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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

ITEMS FROM PAKISTAN

AGRONOMIC RESEARCH STATION

Bahawalpur, Pakistan.

Identifying sources of resistance to wheat leaf rust under induced and natural conditions.

Altaf Hussain Tariq, Saeed Ahmad, Muhammad Arshad Hussain, Muhammad Ziaullah,Lal Hussain Akhtar, and Sabir

Zameer Siddiqi.

Background. Rust diseases pose a major threat to the productivity of wheat crop when epidemics develop. Leaf rust is

world wide in distribution and a most dreaded disease that can spread rapidly and devastate the wheat crop (McIntosh et

al. 1997). In Egypt, Abdel Haq et al. (1980) estimated yield losses up to 50 % in wheat. This disease has appeared in

epidemic form several times in Pakistan. During 1978, a national loss of 86 x 106 USD was estimated (Hussain et al.

1980). Chemical control of rust diseases is not economical. Therefore, cultivation of resistant cultivars is of paramount

importance. Breeders need to plan their hybridization program judiciously in order to produce cultivars with different

genetic backgrounds for resistance to rusts so that any danger of a disease epidemic can be avoided. The present studies

explored new sources for rust resistance in wheat, which will help the breeders in planning future wheat-breeding

programs.

Materials and methods. Local Wheat Diseases Screening Nurseries (LWDSN) comprised of 293 and 346 advanced

wheat lines were planted at Bahawalpur during 2001 and 2002, respectively. Ten commercial wheat cultivars also were

included in the nurseries. The entries, which gave reactions from trace to MRMS at Bahawalpur, and 10 commercial

cultivars also were sown at Kaghan. Each entry was planted in a single 2-m row, 30 cm apart, at both the locations. Two

rows of susceptible checks (Morocco and Local White) were sown repeatedly after every fifth entry and around the

block. The nurseries were inoculated artificially with a spore suspension of leaf rust by injecting, rubbing, and spraying

from the first week of February until 10 March at Bahawalpur during both the years. Kaghan is a summer station about

7,000 ft ASL. Natural rust epidemics occur frequently in this area. The planting at Kaghan was made during the first

week of June. Observations on rust infections were recorded at 10–15 day intervals throughout the growing period at

Bahawalpur and during the end of August at Kaghan. Data were recorded according to the modified Cobb’s scale at both

locations (Peterson et al. 1948). The observations were compared among years and locations to establish the distribution

of rust incidence.

Results and discussion. The

observations of leaf rust on

the 10 commercial wheats

sown at Bahawalpur and

Kaghan in 2001 and 2002

indicated that the intensity of

rust infection during 2001 was

comparatively higher than that

in 2002 at both locations.

Natural infection at Kaghan

was less in 2002 because of

less precipitation throughout

the country during 2002 and

the environmental influence

on the host-pathogen interac-

tion at Kaghan where the

growing season is shorter (80–

90 days) and cooler with a

shorter daylength (Table 1).

Six cultivars, FSD-85, Inqlab-

Table 1. Reaction to the leaf rust pathogen of commercial wheat cultivars at two

different locations in Pakistan during 2001–02. Infection types are listed as TR =

trace, R = resistant, MR = moderately resistant, MS = moderately susceptible, and S

= susceptible.

RARI, Bahawalpur Kaghan

Cultivar (artificial inoculation) (natural infection)

2001 2002 2001 2002

Kohinoor-83 60S 40S 30S 20S

Faisal-85 20R 5R 10MR 5MS

Inqlab-91 TR (< 5 %) 5MR 10MRMS 5MR

Pasban 40S 20S 20S 10S

Rohtas-90 TR 30MRMS 30MRMS 20MRMS

Punjab-96 5MSS 5MS 5MRMS 5MR

Bahawalpur-97 20MRMS 10MRMS 20RMR 10MRMS

MH-97 20MR 20MR 20RMR 5MR

Uqab-2000 10MR 5R 30RMR 20MR

Iqbal-2000 20RMR 30MR 20RMR 5MR

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

91, Rohtas-90, MH-97, Uqab-2000, and Iqbal-2000, were resistant with < 5–30 % infection during both years at

Bahawalpur. At Kaghan, these cultivars exhibited almost the same reaction but with less intensity during 2002. These

cultivars have the Lr10 gene along with Lr27+Lr31 have been very effective in providing resistance to leaf rust. In field

experiments conducted at Faisalabad in Pakistan, Khan et al. (1997) found Pavan, Faisalabad-85, and Inqlab-91 to be

slow rusting. Chaudhry et al. (1996) evaluated 14 commercial wheat cultivars in the field and reported Inqlab-91,

Parwaz-94, and Chakwal-86 resistant to leaf and yellow rust throughout Punjab and the North Western Frontier Province

during 1994 and 1995. Kohinoor-83, Pasban-90, and Punjab-96 remained susceptible to leaf rust at both sites under

induced and natural conditions, whereas Bahawalpur-97 maintained its MR–MS level during both the years.

The leaf rust observations at

the different locations of new ad-

vanced lines during 2001 and 2002

are presented in Table 2. These

observations indicate the number of

test entries under different categories

of rust-infection levels. During 2001,

95 of 293 entries were immune and

135 (46 % of the total) were trace to

moderately resistant. Among 346

lines, 77 remained immune, 92 had

trace infection, 103 were resistant,

and 51 were moderately resistant

during 2002. At Kaghan, the number

of entries was less compared to

Bahawalpur during both years,

because they were selected on the

basis of disease reactions (traces to

resistant and moderately resistant) and

yield traits. Generally, the entries that

were moderately susceptible under

induced conditions at Bahawalpur

were mostly resistant to moderately

resistant reactions at Kaghan during

both years. The inheritance of leaf rust resistance was better in these lines. Rust inoculum is dynamic in nature and

changes from year to year and place to place. Virulence in one environment may not necessarily appear in another

(Khan et al. 2002). The virulence patterns observed at the two sites confirm this hypothesis.

The evolution of new rust races is a permanent feature of the rust pathogen. Whenever new cultivars are

deployed in the field, new races of the pathogen develop after several years and the existing cultivars become suscep-

tible. This phenomenon has been reported by number of workers (Ezzahiri 1989; Meshkova 1990; Meena-Kumari et al.

1992). At present, more than 80 % of the area under wheat cultivation is occupied by the single cultivar Inqlab-91,

which is fraught with the danger. Under these circumstances, steps to avoid monoculture need to be taken. A number of

advanced lines are available from the present studies that were resistant to prevailing rust races to provide sufficient

material for developing new, resistant wheat cultivars.

References.

Abdel Hak ATM, El-Sherif NA, Bassiourny AA, Shafik II, and El-Dauadi Y. 1980. Control of wheat leaf rust by

systemic fungicides. In: Proc Fifth Eur Mediterranean Cereal Rust Conf, Bari, Itlay. Pp. 255-266.

Chaudhary MH, Hussain M, and Shah JA. 1996. Wheat rust scenario, 1994-1995. Pak J Phytopathology 8(1):96-100.

Ezzahiri B and Rolelf AP. 1989. Inheritance and expression of adult plant resistance of leaf rust in Era Wheat. Plant Dis

73:549-551.

Hussain M, Hassan SF, and Kirmani MAS. 1980. Virulence in Puccinia recondita Rob.Ex. Desm. of sp. Tritici in

Pakistan during 1978 and 1979. In: Proc Fifth Eur Mediterranean Cereal Rust Conf, Bari, Itlay. Pp. 179-184.

Khan MA and Hussain M. 2002. Wheat leaf rust (Puccinia recondita) occurrence and shift in its virulence in Punjab

and NWFP. Pak J Phytopathology 14(1):1-6.

Khan MF, Ilyas MB, and Hussain M. 1997. Impact of leaf rust infection on grain yield of various wheat cultivars. Pak J

Phytopathology 9(1):64-66.

Table 2. Reaction to the leaf rust pathogen under natural infection and

induced conditions in new advanced lines at two different locations in

Pakistan during 2001–02. Infection types are listed as I = immune, TR =

trace, R = resistant, MR = moderately resistant, MS = moderately suscep-

tible, S = susceptible, and HS = highly susceptible.

Number of plants

RARI, Bahawalpur Kaghan

(induced epidemic) (natural infection)

Infection type 2001 2002 2001 2002

I (0) 95 77 17 61

TR (< 5 %) 51 92 45 70

R (5–20 %) 70 103 112 114

MR (21–40 %) 14 51 46 20

MS (41–50 %) 28 10 11 6

S (51–80 %) 35 13 1 4

HS (> 80 %) — — — —

Total 293 346 232 275

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

McIntosh RA, Wellings CR and Park RF. 1997. Wheat rusts. An Atlas of resistance genes. CSIRO Publications, P.O.

Box-89, 314 Albert Street, East Melbourn, Victoria 3002, Australia.

Meena-Kumari KVS, Singh DV and Srivastava KD. 1992. Estimation of yield losses in wheat cultivars due to leaf rust

at different growth stages under artificial inoculation. Ind Phytopathology 45(2):266-268.

Meshkova LV. 1990. Source of resistance to wheat brown rust in Western Siberia. Nauch noissledovatel, Skii Institut

Sels Kogo Khozyatstva 6:10-16 and USSR Rev Plant Path 1992, 71(2):802.

Peterson RF, Campbell AB, and Hannah AE. 1948. A diagrammatic scale for estimating rust intensity on leaves and

stems of cereals. Can J Res Sec C 26:496-500.

Performance of advanced wheat genotypes to Helicoverpa armigera Hubner.

Abdul Rashid, Habib Ahmad Saeed, Lal Hussain Akhtar, Altaf Hussain Tariq, and Sabir Zameer Siddiqi.

Background. Wheat is the staple diet of the people of Pakistan, contributing 12.1 % to value added in agriculture and

2.9 % to the GDP. Wheat was grown on an area of 6.30 x 106 ha with a production of 15.42 x 106 tons in 2000–01 in

Punjab (Anonymous 2001). The by-products of wheat are used in bakery products and confectionery. For the last few

years, Pakistan has become self sufficient in wheat production. Surplus wheat is exported to various countries such as

Vietnam, United Arab Emirates, Somalia, Egypt, Ethiopia, Kenya, and Afghanistan. Various rust and smut diseases,

aphids, Helicoverpa armigera, and termites attack this crop. Ann (2002) observed that aphids can be controlled easily

with predators such as Coccinelid beetles and chrysopa and syrphis flies, whereas the reverse is true for H. armigera,

which is a devastating pest of many crop plants world wide (Patankar et al. 2001). Saleem and Rashid (2000) reported a

loss of 13.98 % in grain yield in wheat caused by a single caterpillar of H. armigera per tiller. Being the staple diet, the

use of chemicals is not feasible for the control of this pest because of residual effects that may be hazardous to human

health. The ultimate solution to the problem is the screening of genotypes with built-in resistance to H. armigera.

Keeping in view the significance of the pest, we screened for genotypes of wheat resistant or tolerant to H. armigera.

Materials and methods. To asses wheat losses caused by H. armigera, 20 advance strains of wheat including two

checks were evaluated for spike and grain damage during the Rabi season 1998–99 at the Regional Agricultural Research

Institute, Bahawalpur. The experiment setup was a RCB design with three replications and plot size of 12 m2. Similar

agronomic practices were applied to all genotypes throughout the growing season. Observations of spike damage were

recorded at the harvest by counting the total number of spikes and the number of spikes damaged by the pest from three

randomly selected spots of 1 ft2 from each plot. Grain-damage data were recorded by counting the total number of grains

and number of grains damaged by the pest from five randomly selected spikes from each plot after harvest. Thus, the

percentage of damaged spikes/grains was calculated as follows:

No. of damaged spikes / grains

Spike/grain damage (%) = x 100

No. of total spikes/grains

Data were subjected to statistical analysis using a computer package MSTATC. Correlations were computed

using the Correlation subprogram of

MSTATC. Means were compared by

Duncan’s New Multiple Range Test

(Steel and Torrie 1980).

Results and discussion. Statistical

analysis of the data revealed the highly

significant differences among the mean

values of spike and grain damage (P <

0.01) of all the genotypes (Table 3).

Spike and grain damage ranged from

19.95 to 80.47 and 3.90 to 22.16 % in

the check genotypes, respectively

(Table 4). The most susceptible

genotypes in terms of spike damage

Table 3. Analysis of variance of data with regard to spike and grain damage

of various wheat genotypes after damage by Helicoverpa armigera.

Damaged spikes Damaged grains

Parameter (%) (%)

Means squares 65392 96.35

Probability 0.000 0.000

Coefficient of variation (%) 3.54 % 7.32 %

Cd1 (0.05 %) 2.728 1.558

Cd2 (0.01 %) 3.654 2.086

Standard error 0.953 0.544

Correlation between the two traits (r2) 0.422

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

were D-94654 (80.47 %), PR-68

(76.00 %), WS-94194 (59.53 %),

and V-94091 (58.28 %). The

genotype 92T001 was found to be

the most tolerant with the least spike

damage (19.95 %). For grain

damage, SD-4 had the maximum

damage (22.16 %) and V-8120 had

the least (3.89 %), a vast range of

damage differences. The present

results support the data of Saleem

and Rashid (2000) who found that a

single caterpillar of H. armigera per

tiller caused 13.98 % loss in grain

yield of wheat. Such information

will encourage the wheat breeders to

incorporate this character in their

breeding program. Efforts are being

made to develop the wheat geno-

types tolerant to H. armigera at our

institute.

References.

Anonymous. 2001. Agriculture.

Economic Bulletin, Economic &

Business Research Wing, Na-

tional Bank of Pakistan.

28(8):19-20.

Anonymous. 2002. Agriculture. Economic Bulletin, Economic & Business Research Wing, National Bank of Pakistan.

29(7-8):19-20.

Patankar AG, Giri AP, Harsulkar AM, Sainari MN, Deshpade VV, Ranjekar PK, and Gupta VS. 2001. Complexity in

specificities and expression of Helicoverpa armigera gut proteinases explains polyphagous nature of insect pest.

Insect Biochem Mol Biol 31:453-464.

Saleem M and Rashid A. 2000. Helicoverpa armigera infestation on various wheat varieties. Ann Wheat Newslet

46:101-102.

Steel RGD and Torrie JH. 1980. Principles and Procedures of Statistics. McGraw Hill Book Company, New York, NY.

Pp. 187-188.

Manthar – a high-yielding cultivar of wheat released for general cultivation in southern Punjab.

Sabir Zameer Siddiqi, Mushtaq Ahmad, Manzoor Hussain, Lal Hussain Akhtar, Abdul Rashid, Ghulam Hussain,

Muhammad Aslam, Muhammad Safdar, Muhammad Masood Akhtar, Muhammad Rafiq, and Muhammad Arshad.

Background. Wheat is the main staple food of the people of Pakistan and is grown on the largest area covering more

than 15 x 106 acres in the Punjab. Although Pakistan is a wheat exporter, this situation has been changing for the last 2

years. New steps now are needed to be adopted to progress forward. Agronomic advancement is the utmost need,

including the development of genotypes possessing high-yield potential. Wheat breeders are trying to improve the

potential at their research stations, resulting in wheat cultivars with acceptable and improved characteristics.

Manthar is selection from CIMMYT material and has been tested at Regional Agricultural Research Institute,

Bahawalpur and outstations for 7 years. This strain has the famous CIMMYT line Kauz in its pedigree, which is a more

adapted and a high yielder. Genetically, this strain differs from existing commercial cultivars of Punjab. Manthar rated a

position among the top five strains in National Uniform Wheat Yield Trial the first year and the first position in late

planting and second in 23 sites in Pakistan in its second year. Manthar has improved yield potential and better adaptabil-

Table 4. Data for various traits of the wheat genotypes tested for resistance to

Helicoverpa armigera at the Regional Agricultural Research Institute,

Bahawalpur during 1998–99.

Genotype Damaged spikes Damaged grains Yield (kg/ha)

V-95219 41.01 11.48 4,062

94B-3047 39.34 14.19 3,861

WS-94194 59.53 15.18 3,674

V-94105 52.37 9.58 3,861

PR-68 76.00 14.94 3,243

D-94654 80.47 18.09 3,292

SD-4 39.04 22.16 3,049

92T001 19.95 4.09 4,035

V-95153 45.75 20.52 4,021

AUP-9701 34.58 6.12 4,333

V-94091 58.28 11.44 3,597

93B2707 37.85 14.32 3,674

PR-67 46.72 9.01 3,507

V-95069 36.22 12.29 3,604

DN-10 50.48 9.96 2,931

V-8120 24.23 3.90 3,382

91BT010-1 45.33 4.00 3,986

V-94045 52.42 18.24 3,326

INQ-91 52.37 19.78 3,674

Local check 41.31 19.12 3,771

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

ity. Dry and unfavorable conditions in 2001–02 produced a successful wheat crop during a continuous drought. This

genotype rated the second position in Pakistan based on drought and heat tolerance. The cultivar is resistant to leaf rust

and yellow rust at CDRI, Islamabad. We hope that Manthar will help boost the average wheat yield because of its better

economic characteristics while being a general-purpose cultivar.

Materials and methods. Manthar, selected from CIMMYT (Mexico) material, was tested at Regional Agricultural

Research Institute, Bahawalpur, and outstations for 7 years (1996–

2002) and given the number 97B2210. This line was evaluated for

its yield potential in 81 trials at various locations Preliminary

Yield and Advanced Yield Trials, the Micro Wheat Yield Trials

(2000–01), and the National Uniform Wheat Yield Trial (2000–

01). Sowing date and fertilizer trials also were conducted to

evaluate its production technology during 2000–01 to 2001–02.

The line 97B2210 also was tested for resistance to rusts, loose

smut, and Karnal bunt at the Regional Agricultural Research

Institute, Bahawalpur; the Wheat Research Institute, Faisalabad,

and the Crop Disease Research Institute, NARC, Islamabad during

2000–02 and compared with standard cultivars. The Coordinator

Wheat, NARC, Islamabad, also studied the quality characteristics

of the line in 2000–01. The Federal Seed Certification and

Registration Department, Islamabad, evaluated plant characteris-

tics. The yield data were subjected to ANOVA using the MSTAT

statistical program and means were compared using Duncan's

Multiple Range Test (Steel and Torrie 1980).

Results and discussion.

Yield performance. Station Yield Trials. Manthar was tested in

preliminary and advance yield trials at the Regional Agricultural

Research Institute, Bahawalpur, between 1996–97 and 2001–02 in

late planting and compared with the national checks, Uqab-2000

and Inqlab-91. The performance of Manthar is given in Table 5.

Over a 3-year average, the cultivar had a 7.1 % higher yield than

Inqlab-91 (Table 5) and also outyielded the check by a margin of

3.7 % in zonal trials conducted at three

locations in 1999–2000 (Table 6).

Micro Wheat Yield Trial. The

Director, Wheat Research Institute,

Faisalabad, also evaluated the perfor-

mance of Manthar under a coded number

during 2000–01 at various locations in

Punjab in replicated yield trials. The

results show yields 2.0, 13, and 11 %

higher for Manthar when compared to

Inqlab-91, Uqab-2000, and Iqbal-2000,

respectively, an average of 10 locations

(Table 7).

National Uniform Wheat Yield

Trial. The Coordinator Wheat,

Islamabad, also evaluated Manthar in a

replicated trial called the National

Uniform Yield Trial under normal and

short conditions throughout Pakistan

during 2001–02. The performance of

Table 5. Results of the station yield trials at

Bahawalpur, Pakistan, for Manthar (97B2210)

and the check cultivar Inqlab-91.

Year Trial 97B2210 Inq-91

1997–98 A1 (N) 5,671 a5,322 a

1998–99 B3 (N) 4,750 a4,417 b

1999–2000 CI (N) 6,115 a5,693 b

Average 5,512 5,144

% increase over check + 7.1

Table 6. Zonal testing of Manthar (97B2210) and

the check cultivar Inqlab-91 (Inq-91) at three

locations in Pakistan during 1999–2000.

Location 97B2210 Inq-91

CRSS, Haroonabad 5,245 4,936

ORS, Khanpur 4,442 4,393

ARS, Khanewal 4,782 4,630

Average 4,823 4,652

% increase over check + 3.7

Table 7. Results of the Micro Wheat Yield Trials at various locations in

Pakistan in 2000–01. Source: Director Wheat, Faisalabad.

97B2210 Uqab Iqbal

Location (Manthar) Inqlab-91 2000 2000

RARI, Bhawalpur 5,405 a4,826 a5,004 a4,676 b

ARF, Rahim Yar khan 5,204 a4,932 a4,721 b4,186 bc

CRSS, Haroonabad 6,346 a5,990 a3,741 c4,486 bc

WRL, Faisalabad 5,735 a5,920 a5,965 a5,550 a

ARF, Vehari 3,290 ab 3,660 a3,382 ab 3,290 ab

PSC, Khanewal 3,799 b4,819 a4,819 a5,097 a

Thatta Jawana Jhang 4,263 a3,614 b4,031 a4,031 a

Hafizabad Pindi Bhattian 4,170 a4,263 a2,124 c2,965 b

ARF, Gujranwala 4,911 a4,726 a4,355 b4,633 a

RRI, Kala Shah Kaku 4,720 a4,165 b4,165 b4,165 b

Average with PSC 4,784 4,691 4,231 4,307

% increase over check + 2 + 13 + 11

Average without PSC 4,894 4,677 4,165 4,220

% increase over check + 4.66 + 18 + 16

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Manthar in this trial is given is Table 8. Manthar

had a 7.1 % higher yield than the local check at the

National level on the basis of 12 locations in 24

trials.

Varietal characteristics. Various varietal

characteristics recorded by the Federal Seed

Certification and Registration Department,

Islamabad, in comparison with Inq-91 are given in

Table 9.

Agronomic studies. Six trials were

conducted at Regional Agricultural Research

Institute, Bahawalpur, during 2000–02 to ascertain

production technology. Sowing time is 10

November to 10 December at a seeding rate of 125

kg/ha. Fertilizer requirements include 125–100–

50 NPK with 4–5 irrigations.

Pathology studies. The response of

Manthar to various foliar diseases was studied at

Crop Diseases Research Institute, NARC,

Islamabad; the Wheat Research Institute,

Faisalabad; and the Regional Agricultural Re-

search Institute, Bahawalpur. The data are given

in Table 10. The data indicates that Manthar is

resistant/tolerant to the yellow rust, leaf rust, loose

smut, Fusarium, and Karnal bunt pathogens.

Entomology studies. The response of

Manthar to aphid and Helicoverpa armigera also

was studied at Regional Agricultural Research

Institute, Bahawalpur, in 2000–02. Data are given

in Table 11 shows the performance of Manthar as

compared to commercial checks.

Quality studies. The quality characters

were recorded by the National Agricultural

Research Centre, Islamabad, and are given in

Table 12. The new cultivar is

better than the existing checks.

Conclusion. The cultivar

Manthar (97B2210) not only is

a high-yielder and tolerant/

resistant to all diseases but also

best suited to a wheat–cotton–

wheat rotation. Because of

better adaptability, Manthar has

the potential of replacing

previously approved wheat

cultivars, especially in the

southern Punjab. This cultivar

was approved and released by Variety Evaluation Committee, Islamabad, for general cultivation during 2002.

Table 10. Disease response of Manthar and a local check to rust recorded by the

Crops Disease Research Institute, Islamabad, during 2000–01.

ACI RRI

Year Cultivar leaf rust yellow rust leaf rust yellow rust

2000–01 97B2210 3.4 — 6.7 —

Local White 56.6 — — —

2001–02 97B2210 0.7 0 7.6 8.9

Local White 45.65 — — —

Table 9. Ccharacteristics of Manthar compared to the

local check cultivar Inqlab-91.

Characteristic Manthar Inqlab-91

Days to handing 98 114

Days to maturity 142 135

Plant height 94 cm 98 cm

Lodging Resistant Resistant

Tillers per meter row 145 132

1,000-kernel weight 40–45 g 44.0 g

Protein 12.97 % 10.51 %

Disease reaction Resistant/tolerant Resistant

Grain size Medium —

Maturity status Medium Medium

Growth habit Erect Drooping

Yield potential 6,708 kg/ha 6,900 kg/ha

Table 8. Results of the National Uniform Wheat Yield Trial at

various locations in Pakistan in 2001–02. Seeding date is for

normal and late dates combined. Source: Anonymous 2002.

97B2210

Location (Manthar) Local check

ARF, Rahim Yar khan 3,773 3,348

ORS, Khanpur 4,081 3,619

RARI, Bahawalpur 3,583 3,335

CRSS, Haroonabad,BWN 3,827 3,490

ARF, Vehari 3,852 3,583

PSC, Khanewal 3,919 4,177

WRI, Faisalabad 4,843 4,853

ARF, Layyah Karore 2,977 2,125

Gill Model Farm S.Abad Jhang 3,700 3,382

Hafizabad Pindi Bhattian 4,344 4,567

In service Trg. Sargodha 3,927 3,281

ARF, Sheikhpura 3,813 3,792

Average 3,887 3,629

% increase over check + 7.1

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

References.

Anonymous. 2002. National Uniform Wheat Yield Trials

Summary Results 2001-2002. National Agricultural

Research Centre, Islamabad.

Mustafa SZ, Yasmin S, and Kisana NS. 2001. Results of

the National Uniform Wheat Yield Trials, 2000–2001.

Coordinated Wheat-Barley and Triticale Programme,

Pakistan Agricultural Research Council, P.O. Box 1031,

Islamabad. P. 25.

Steel RGD and Torrie JH. 1980. Principles and Procedures

of Statistics. McGraw Hill Book Company, New York.

Pp. 187-188.

Development of 012679, a new wheat strain with

special characteristics.

Table 11. Leaf rust reaction of Manthar (97B2210) and a check in the National

Wheat Disease Screening Nursery at CDRI, Islamabad, 2001–02.

PRC, AARI, RARI, CCRLD, NIFA, NARC, CDRI RRI

Cultivar SKT FSD BWP SBK PWAR ISD KHI

97B2210 0 10MR 0 0 0 5MRMS 0 7.6

Morocco 50S 90MS 50MSS 40S 20S 80S 30S —

Table 12. Resistance to aphids in Manthar compared with

the standard commercial check cultivars.

Aphid Yield

Year Cultivar population (kg/ha)

2000–01 Manthar 21.4 3,250

Inq-91 22.3 3,084

2001–02 Manthar 0.50 2,512

Auquab-2000 0.55 2,392

Iqbal-2000 0.55 2,332

Table 14. Results of the National Uniform Wheat yield

Trial in 2000–01 for Manthar compared to the local

check cultivar Inqlab-91.

Characteristic Manthar Inqlab-91

1,000-kernel weight 42.3 g 37.0 g

Test weight 79.5 g 74.2 g

PSI (%) 29.0 42.2

Ash (%) 1.55 1.54

Gluten content MS MS

Dry gluten (5) 8.20 5.79

Crude protein (%) 12.79 10.06

Table 13. Resistance to Helicoverpa armigera in

Manthar and some commercial check cultivars in 2000–01

Aphid population Yield

(per tiller) (kg/ha)

Cultivar Normal Late Norma Late

Manthar 0 0.30 4,475 4,175

Inqbal-91 0.33 0.62 4,150 3,880

MH-97 0.34 7.11 4,262 3,925

Mushtaq Ahmad, Ghulam Hussain, Muhammad Rafiq, Manzoor Hussain, Lal Hussain Akhtar, and Sabir Zameer Siddiqi.

Wheat not only is the main staple food of Pakistan, but more than 33 % population of world also depend upon it for

nourishment. Hybridization efforts are not bearing significant yield improvements. Improvement in grain yield is the

ultimate objective of all agronomic and breeding investigations. Genetic yield potential can be improved by increasing

the number of grains/unit area and grain weight. Efforts at the Regional Agricultural Research Institute, Bahawalpur,

seek to improve grain weight and grain number/unit area and combine them in the same plant with required protein and

gluten levels. A new wheat strain was bulked during 2000–01 with number 012679. Strain 012679 is a local cross

(Debaria/WL-711) attempted during 1994–95. The F1 to F6 were grown from 1995–96 to 2000–01 at RAI, Bahawalpur.

The cultivar was evaluated for yield in yield trials during 2001–02 with under the number 012679.

Strain 012679 produced 41.12 % and 61 % more yield than the commercial checks Inqbal-91 and PND-I,

respectively (Tables 15 and 16). Further studies are in progress in yield trials during 2002–03 to confirm these results.

Strain 012679 differs from the existing cultivars in following characteristics: a thick stem is resistant to lodging;

increasing the seeding rate compensates for a lower number of tillers/seed; early maturity fits in a wheat-based cropping

pattern; a thick, dense head with 100 % maturity gives more grains/spike; and more grains than commercial standards

results in a higher grain yield.

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Effect of irrigation and various nitrogen

and phosphorus levels on wheat yield.

Muhammad Aslam, Manzoor Hussain, Lal

Hussain Akhtar, Mushtaq Ahmad, Ghulam

Hussain, Abdul Rashid, Muhammad Safdar,

Muhammad Masood Akhtar, Muhammad Arshad,

and Sabir Zameer Siddiqi.

Background. Wheat, a major food grain of

Pakistan, is being adversely affected by shortage

of water. During 2001–02, a decline of 2.4 % in

cultivated area and yield was found due mainly to

dry weather, a shortage of irrigation water, low

application of NP fertilizer, and a delayed sowing

of the 2001–02 season crop (Sabir et al. 2000;

Anonymous 2002). Under these circumstances,

the positive role of irrigation and NP levels need to

be demonstrated. Similarly, the high use of

irrigation water also is being restricted due to

shortage of canal water and high prices of subsoil

water. The NP fertilizer and irrigation factors play

an important role in getting the highest grain yield

from the wheat crop. Ibrahim (1999) obtained a

high grain yield of 4.6 and 4.8 t/ha using three and

four irrigations. Kalita et al. (2000) achieved a high grain yield from three irrigations. Laxminarayana and Thakur

(1999) found that grain yield increased with an increase in applied phosphorus up to 90 kg/ha. Sabir et al. (2000)

obtained their highest yields with the application of 150:100 kg/ha N:P. Pandey et al. (1999) reported that grain yield

increased up to 150:75 N:P levels. Naser et al. (1999) and Maliwal et al. (2000) found that irrigation treatments increase

the yield. Therefore, this project was to determine the best NP level with three and five irrigations for obtaining best

wheat yield.

Materials and methods. The study was conducted at Regional Agricultural Research Institute, Bahawalpur, during the

years, 2000–02. The wheat cultivar Punjnad-1 was sown during both the years on well prepared seed bed with a single-

row drill in rows 30 cm apart. Ten treatments involving two irrigation levels (three (at crown root, boot, and milk stages)

and five (at crown root, tillering, boot, milk, and grain-formation stages)) with five levels of NP 0–0, 50–50, 100–75,

150–100, and 200–125 kg/ha, were studied. K was kept constant (60 kg/ha) in all treatments. A split-plot design with

four replications was used with net plot size of ‘6 m x 1.8 m’. All phosphorus and potassium was applied as a basal dose

at sowing. All nitrogen fertilizer was applied with the first irrigation. Other agronomic practices were kept uniform for

all the treatments. Grain yield

(kg/ha) was recorded at

harvest. The data were

analyzed statistically by using

Fisher’s analysis of variance

and differences among the

treatments means were

compared by Duncan’s

Multiple Range Test (Steel

and Torrie 1980). Table 17

lists the treatments given.

Results and discussion. Grain yield significantly increases with interactive effects of irrigation and NP (Table 18). T4

gave the highest grain yield of 3,678 kg/ha, which was more economical than T5 because addition of 50–25 kg/ha more

NP in T5 compared to T4 resulted in only 144 kg/ha additional yield which is uneconomical. T7 gave four times more

yield (2,360 kg/ha) than T6 (558 kg/ha). Similarly T8 and T9 gave maximum yield of 3,983 and 4,178 kg/ha, respec-

tively. T10 was at par with T9. The present results support the findings of E1-Far and Teama (1999) who reported that

Table 17. Different treatment regimes used in evaluating different nitrogen and

phosphorus levels and irrigation levels on wheat yield.

NP (kg/ha)

Irrigation 0–0 50–50 100–75 150–100 200–125

Three T1 T2 T3 T4 T5

Five T6 T7 T8 T9 T10

Table 15. Yield data for the new cultivar 012679 compared

with commercial cultivars in 2001–02 at the Regional Agricul-

tural Research Station, Bahawalpur, Pakistan.

Yield Yield

Cultivar (kg/ha) Cultivar (kg/ha)

012672 3,953 012678 4,848

012673 5,222 012679 5,796

012674 4,710 012680 5,219

012675 4,538 Inq-91 4,108

012676 3,810 Punjnad-I 3,600

012677 5,067 Uquab-2000 3,545

Table 16. Yield components of the new cultivar 012679

compared to the local checks in 2001–02 at the Regional

Agricultural Research Station, Bahawalpur, Pakistan.

1,000-kernel No. of Spike Yield

Cultivar weight (g) grain/spike weight (g) (kg/ha)

012679 50.05 108 5.24 5,796

Inqbal-91 40.45 55 4.32 4,108

PND-I 38.20 59 4.54 3,600

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

grain yield was the highest when crop

was irrigated after every 31 days and

lowest when irrigation was applied

after every 60 days. Ibrahim (1999)

obtained grain yield of 4.6 t/ha and

4.8 t/ha using three and four irriga-

tions, respectively. Kalita et al.

(1999) obtained the highest grain

yield from three irrigations. Pandey

et al. (1999) reported that grain yield

increased up to 150:75 kg/ha NP.

Sabir et al. (2000) obtained the

highest yield with the application of 150–100 NP. Laxminarayana and Thakur (1999) reported that grain yield increased

with increase of phosphorus upto 90 kg/ha. Five irrigations were applied at crown root, tillering, boot, milk, and grain-

formation stages.

References.

Anonymous. 2002. Agriculture: wheat production below target. Economic Bulletin, National Bank of Pakistan. 29(11-

12):16.

Ibrahim SM. 1999. Wheat cultivation under limited irrigation and high water table conditions. Egypt J Soil Sci

39(3):361-372.

Kalita MC, Sarmah NN, and Barkakoty PK. 1999. Irrigation regimes on growth and yield of wheat under different land

situations. J Agric Sci Soc North East India. 12(1):18-23.

Laxminarayana KI and Thakur NSA. 1999. Effect of phosphorus on yield performance of wheat in acidic soils of

Mizoram. J Hill Res 12(2):138-140.

Maliwal GL, Patel JK, Kaswala RR, Patel ML, Bhatnagar R, and Patel JC. 2000. Scheduling of irrigation for wheat

(Triticum aestivum) under restricted water supply in Normada region. Ind J Agric Sci 70(2):90-92.

Naser HM, Islam MT, Begum HH, and Idris M. 1999. Effects of time and frequency of irrigation on yield of wheat.

Thai J Agric Sci 32(2):205-209.

Pandey AK, Chauhan VS, Prakash V, and Singh RD. 1999. Seed and fertilizer management under non-availability of

irrigation at crown root initiation stage in wheat. Crop Res (Hisar) 17(3):286-291.

Sabir MI, Ahmad I, Shah SAH, and Shahzad MA. 2000. Effect of different rates and NP levels on growth and yield of

wheat (Triticum aestivum). J Agric Res 38(4):311-317.

Steel RGD and Torrie JH. 1980. Principles and Procedures of Statistics. 2nd Ed. McGraw Hill Book Co. Inc.

Singapore. Pp. 172-177.

Effect of irrigation at different growth stages on the grain yield of wheat.

Muhammad Aslam, Manzoor Hussain, Lal Hussain Akhtar, Abdul Rashid, Ghulam Hussain, Muhammad Safdar,

Muhammad Arshad, and Sabir Zameer Siddiqi.

Background. Wheat is the most important Rabi cereal crop of Pakistan. Because of deficits in irrigation water in the

rivers, the country is facing long-lasting moisture stress. A plan that utilizes our limited sources of irrigation water in

such a way that country does not suffer food shortage is needed. Ibrahim (1999) obtained grain yields of 4.3, 4.6, and

4.8 t/ha by applying 2, 3, and 4 irrigations, respectively. Naser et al. (1999) obtained the highest yield with two irriga-

tions applied 30 and 50 days after sowing. Kalita et al. (1999) obtained high grain yields from three irrigations. Similar

results have been reported by Lidder et al. (1999), Tripathi et al. (2000), and El-far and Teama (1999). The present study

was planned to define the critical stages of the wheat crop using limited number of irrigation water to obtain an optimum

yield.

Materials and methods. The study involved 15 treatments laid out in a RCBD with three replications (Table 19). Net

plot size was ‘6 m x 1.8 m’. The wheat cultivar Punjnad-I was sown during the first week of December 2000–02.

The recommended fertilizer dose was applied to all the treatments. Punjnad-I was sown during both years on a

well-prepared seed bed with a single-row hand drill in rows 30 cm apart. All other agronomic practices were kept

Table 18. Grain yield in Punjand-1 wheat under various treatment regimes

varying in level of nitrogen and phosphorus fertilizer and number of irriga-

tions.

NP (kg/ha)

Irrigation 0–0 50–50 100–75 150–100 200–125

Three 474 F1,482 E3,383 C3,678 BC 3,822 ABC

Five 558 F2,360 D3,983 A4,178 A4,082 A

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

uniform for all treatments. Grain yield/ha was

recorded at harvest. The data were analyzed

statistically using Fisher’s ANOVA and differences

among the treatment means were compared by

LSD (Steel and Torrie 1980).

Results and discussion. One irrigation. One

irrigation was applied at different four growth

stages of wheat crop. Irrigation applied at boot

stage gave the maximum yield compared to other

stages (Table 20). Similar results were reported by

Ibrahim (1999).

Two irrigations. Two irrigations were

applied in six of the combinations. Treatment T10

(boot + milk; 2,676 kg/ha) gave the highest yield

of these treatments. Ibrahim (1999), Naser et al.

(1999), and Lidder et al. (1999) also achieved best

results when irrigation was applied at similar

stages.

Three irrigations.

Three irrigations were applied

in three combinations. Irriga-

tions applied at crown root +

boot + milk stages (T12) gave a

maximum yield of 3,200 kg/ha.

These results are in line with

those of Ibrahim (1999),

Maliwal et al. (2000), Naser et

al. (1999), and Lidder et al.

(1999) who studied similar

growth stages for irrigation and

found that three irrigation

applied at above-mentioned

stages gave the best yield.

Four and five

irrigations. Four and five

irrigations were applied

according to the tradition in the

southern Punjab. Yields of

3,987 and 4,139 kg/ha were

recorded for four and five

irrigations, respectively. Grain

yield declines of 55.6–69.6, 35.4–51.2, 22.7–33.0, and 3.7 % were observed using 1, 2, 3, or 4 irrigations, respectively,

compared to five irrigations. The results are supported by the findings of Naser et al. (1999) and Lidder et al. (1999)

who studied various numbers of irrigations at various growth stages and found that all irrigation treatments increased

yield.

Table 19. Wheat growth stages used to assess the effect of

irrigation for optimum yield.

1. Crown root

2. Tillering

3. Boot

4. Milk

5. Crown root + tillering

6. Crown root+ boot

7. Crown root+ milk

8. Tillering + boot

9. Tillering + milk

10. Boot + milk

11. Crown root + tillering + boot

12. Crown root + boot + milk

13. Tillering + boot + milk

14. Crown root + tillering + boot + milk

15. Crown root + tillering + boot + milk + grain formation

Table 20. Grain yield in Punjnad-I wheat with irrigations applied at various growth

stages.

Irrigations applied at Grain yield (kg/ha)

1. Crown root 1,260 hi

2. Tillering 1,433 ghi

3. Boot 1,836 fgh

4. Milk 1,494 ghi

5. Crown root + tillering 2,018 defg

6. Crown root + boot 2,620 cde

7. Crown root + milk 2,018 efg

8. Tillering + boot 2,273 def

9. Tillering + milk 2,200 def

10. Boot + milk 2,676 cde

11. Crown root + tillering + boot 2,776 cd

12. Crown root + boot + milk 3,200 c

13. Tillering + boot + milk 2,812 cd

14. Crown root + tillering + boot + milk 3,987 b

15. Crown root + tillering + boot + milk + grain formation 4,139 a

Cd1=666.7 Cd2=921.21

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Conclusion. Depending on the amount of irrigation water available, the best growth stage for application of available

irrigation water include:

1 irrigation Boot

2 irrigations Boot + milk

3 irrigations Crown root + boot + milk

4 irrigations Crown root + tillering + boot + milk

5 irrigations Crown root + tillering + boot + milk + grain formation

References.

El-Far IA and Teama EA. 1999. Effect of irrigation intervals on productivity and quality of some bread and durum

wheat. Assiut J Agric Sci 30(2):27-41.

Ibrahim SM. 1999. Wheat cultivation under limited irrigation and high water table conditions. Egypt J Soil Sci

39(3):361-372.

Kalita MC, Sarmah NN, and Barkakoty PK. 1999. Irrigation regimes on growth and yield of wheat under different land

situations. J Agric Sci Soc North East India 12(1):18-23.

Lidder RS, Jain MP, and Khan RA. 1999. Effect of irrigation and fertility levels on wheat (Triticum aestivum) cultivars

in deep vertisol. Internat J Trop Agric 17(1/4):131-134.

Maliwal GL, Patel JK, Kaswala RR, Patel ML, Bhatnagar R, and Patel JC. 2000. Scheduling of irrigation for wheat

(Triticum aestivum) under restricted water supply in Normada region. Ind J Agric Sci 70(2):90-92.

Naser HM, Islam MT, Begum HH, and Idris M. 1999. Effects of time and frequency of irrigation on yield of wheat.

Thai J Agric Sci 32(2):205-209.

Steel RGD and Torrie JH. 1980. Principles and Procedures of Statistics. 2nd Ed. McGraw Hill Book Co. Inc.

Singapore. Pp. 172-177.

Tripathi P, Tomar SK, and Adhar S. 2000. Effect of moisture regimes and genotypes on biomass accumulation, radiation

interception and its use in wheat (Triticum aestivum). Ind J Agric Sci 70(2):97-101.

Wheat yield potential—current status and future research strategies in Pakistan.

Muhammad Sarwar Cheema, Muhammad Akhtar, and Liaquat Ali.

Wheat is the staple food for most of the people of Pakistan, and wheat straw is an integral part of the daily rations for

livestock. The cultivation of wheat has spread throughout the four provinces of Pakistan. The wheat-growing area and

production for the year 1999–2000 were 73 % and 78 %, respectively for the province of Punjab, with smaller amounts

in the Sindh (13.5 % and 14.5 %), Northwest Frontier (9.5 % and 5 %), and Baluchistan (4 % and 2.5 %) provinces.

Yield potential. A substantial yield gap has been observed at the experimental stations, progressive growers, and on

farmer’s fields in each province. Six, high-yielding wheat cultivars were sown at three different locations in D.I. Khan,

(Northwest Frontier Province), Pakistan, to explore their yield potential under prevailing climatic conditions. Daman 98

ranked first among all the tested cultivars by producing a grain yield of 12.5 t/ha (Khan et al. 2000). Choudhary and

Mehmood (1998) obtained a maximum grain yield of 7 t/ha with Inqlab-91. Sadiq and Khan (1994) also reported 7 t/ha

yield from Pak 81 in a study on the effects of intercropping and planting pattern on yield and yield components of wheat.

Bajwa et al. (1993) reported the influence of different irrigation regimes on the yield and yield components of the wheat

Pak 81, obtained maximum yield of 6.5 t/ha after the application of four irrigations.

Current status. Pakistan’s average grain yield ranged between 2,053 to 2,490 kg/ha over the last 5 years, 1996–97 to

2000–01 (Table 21). A huge yield gap lies between the experimental yield and the average yield of the country, so there

is great hope for double and even triple the wheat grain yield.

Population and wheat requirements. For 2001, the projected population for Pakistan is estimated to be 140.47 x 106

and wheat production is 19.02 x 106 tons. Domestic consumption requirements have been estimated at 20 x 106 tons.

Pakistan became self-sufficient in wheat by producing 21.08 x 106 tons during the year 1999–2000, which was primarily

due to 25 % increase in support price of wheat. Wheat growers produced about one million tons of surplus wheat grain,

a marginal self sufficiency that can be changed at any time by natural hazards. Therefore, concerted efforts are needed to

maintain increased production that meets future requirements.

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Yield gap. A substantial yield gap has been observed between yield at the experimental stations and in farmers’ fields in

each province. This gap is primarily because of the lack of finances on the part of the majority of farmers for imple-

menting modern technology for wheat production. Thus, we hope for improving wheat production and yield in the

country.

Constraints to production. Like many developing countries, wheat production is confronted with both biophysical

constraints (disease, fertilizer, water, seed, cultivars, cultural practices, and salinity/sodicity) and socioeconomic con-

straints (credit, knowledge, experience, tradition, and institutions.).

Disease. Although several diseases attack wheat, stripe and leaf rusts, loose and flag smuts, Karnal bunt, powdery

mildew, Helminthosporium leaf spots, and foot and root rots are the most important in Pakistan. Other diseases, such as

those caused by Septoria spp., downy mildew, black point, and black chaff, are of minor importance. Breeding programs

try to develop wheat cultivars that are resistant or tolerant to these principal diseases. Measures to minimize their

adverse effects on production also are being investigated.

Insect pests. Fortunately, wheat in not attacked by any serious pests, however, infestations of army worm, Hilothus, and

green aphids have occurred in localized areas.

Drought. About 21 % of total wheat area in Pakistan is rainfed. The screening of plant material and the testing of new

cultivars for drought tolerance are made in rainfed areas or under simulated moisture stress. Some cultivars (Inqlab 91,

Punjnad1, and Iqbal 2000) that were developed for irrigated areas also have proven to be very successful under rainfed

conditions. The testing of new cultivars under both irrigated and rainfed conditions is encouraged.

Salinity/sodicity. At present, 2.4 x 106 ha of land in Pakistan have been rendered saline-sodic. With the continuous use

of low-quality water, this menace increases every year. Wheat yield was found to be reduced by 19 % under moderately

saline-sodic soils.

Lack of nutrients. Experiments on yield constraints in irrigated and rainfed areas indicate that the proper application of

fertilizer is of utmost importance. Yield reductions ranging from 50 to 75 % have been observed without proper fertilizer

use and clearly demonstrate that wheat yields can be substantially increased if fertilizer use is properly regulated in the

country.

Table 21. Area, production, and average yield of the wheat crop in the different provinces of Pakistan

between 1996–97 and 2000–01. Units are for area (x 103 ha), production (x 103 tons), and yield (kg/ha).

Source: 2002 Pakistan Statistical Year Book, Agricultural Statistics of Pakistan, Government of Paki-

stan, Islamabad, pp. 114. NWFP is the Northwest Frontier Province.

Province

Year Pakistan Punjab Sindh NWFP Balochistan

1996–97 Area 8,109.1 5,839.9 1,106.8 842.8 319.6

Production 16,650.5 12,371.0 2,443.9 1,064.4 771.2

Yield 2,053.0 2,119.0 2,208.0 1,263.0 2,413.0

1997–98 Area 8,354.6 5,934.6 1,120.2 918.1 381.7

Production 18,694.0 13,807.0 2,659.4 1,356.0 871.6

Yield 2,238.0 2,326.0 2,374.0 1,477.0 2,283.0

1998–99 Area 8,229.9 5,934.6 1,123.7 857.6 314.0

Production 17,857.6 13,212.0 2,675.1 1,221.8 748.7

Yield 2,169.0 2,227.0 2,381.0 1,425.0 2,384.0

1999–00 Area 8,463.0 6,180.3 1,144.2 806.5 332.0

Production 21,078.6 16,480.3 3,001.3 1,067.8 529.2

Yield 2,490.0 2,667.0 2,623.0 1,324.0 1,594.0

2000–01 Area 8,180.8 6,255.5 810.7 790.3 324.3

Production 19,023.7 15,419.0 2,226.5 164.0 614.2

Yield 2,325.0 2,465.0 2,476.0 967.0 1,893.0

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Planting date. More than 50 % of the wheat in Pakistan is planted late, i.e., during the month of December. Planting

date experiments have shown that yield is progressively reduced with delayed planting. Yield was found to be reduced

by 28.8 and 75.8 % when sowing was delayed from November to December and from November to January, respec-

tively,

Weeds. With the introduction of high-yielding and fertilizer-responsive Mexican wheat cultivars during mid 1960s,

weed populations have increased tremendously causing considerable losses in crop yield. No data regarding yield losses

due to weeds is available, however, depending upon the degree of infestation, losses yield are estimated to be between

13–42 %. A number of weed species infest the wheat fields; both grasses and broadleaf weeds. Wild oat (Avena fatua),

canary grass (Phalaris minor), Chenopodium spp., and Convolvulis arvensis has been found to be the major weeds.

When weeds were controlled by the herbicides Dicuran M.A., Tribunil, Graminon, and Arelon, yield increases of 41, 22,

22, and 25 %, respectively, over the weedy controls were found (Ahmed et al. 1987).

Future research strategies. Future strategies for the improvement of wheat will involve more emphasis on breeding

cultivars that possess wider adaptation and can withstand various types of stress (disease, high temperature, cold and

frost, drought, salinity/sodicity, and water logging). Efforts also will be made to develop wheat cultivars with low input

requirements. Improving grain characteristics and milling and baking quality of wheat also will receive greater attention.

References.

Ahmad S, Cheema ZA, Bashir S, and Iqbal M. 1987. Weed management in wheat. Progressive Farming 7(1):33-36.

Anonymous. 2002. Pakistan Statistical Year Book. Agricultural Statistics of Pakistan, Government of Pakistan,

Islamabad. Pp. 114.

Bajwa MA, Chaudhary MH, and Sattar A. 1993. Influence of different irrigation regimes on yield and yield components

of wheat. Pak J Agric Res 14:360-365.

Chaudhary AU and Mehmood R. 1998. Determination of optimum level of fertilizer nitrogen for varieties of wheat.

Pak J Biol Sci 4:351-353.

Khan MA, Hussain I, and Baloch MS. 2000. Wheat yield potential - current status and future strategies. Pak J Biol Sci

3(1):82-86.

Sadiq M and Khan HKA. 1994. Effect of intercropping and planting patterns on yield and yield components of wheat.

Sarhad J Agric 10:351-354.

ITEMS FROM ROMANIA

S.C.A. — AGRICULTURAL RESEARCH STATION

Turda, 3350, str. Agriculturii, 27, jud. Cluj, Romania.

Yield stability and breeding for adaptation in winter wheat.

V. Moldovan, Maria Moldovan, and Rozalia Kadar.

Winter wheat provides a substantially larger amount of the world’s wheat production than does spring wheat because

winter wheat is more productive in those areas where both types can be grown. Thus, winter wheat usually is preferred

over spring wheat in the regions where the climate permits production. The limits of winter wheat adaptation are

established primarily by winter temperature. Thus, the winter survival temperature determines the northern limit and the

winter temperature that is sufficiently low to permit vernalization gives the southern limit of the cropping area. From

this point of view, winter wheat cultivars must have a high enough winter hardiness in northern regions and low require-

ments for vernalization in southern regions to be acceptable to producers.

Improved cultivars substantially contribute to increase wheat production. However, wheat yields in most

production regions seem to be no more than one-half of the potential yield of the cultivars and far below the theoretical

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

maximum yields. This difference reflects powerful production constraints that prevent the true genetic potential for yield

to be expressed by the grown cultivars.

Although wheat-breeding programs have some priorities in common, the major objective of increasing the

genetic potential of yield for most, if not for all, can be achieved via breeding for higher yield potential or by diminishing

or eliminating hazards that reduce yield. Actually, wheat breeding seeks to remove yield constraints by developing

cultivars with resistance to disease, insects, lodging, cold, heat, and drought. Other yield constraints can be best dealt

with through improved cultural practices and management. Obviously, some yield constraints are fixed by the environ-

ment and cannot be manipulated.

As a breeding objective, yield represents an extreme example of a quantitative trait being polygenically inher-

ited and subject to environmental influence to a large extent. Studies have shown that the environmental variation

associated with yield often exceeds genotypic variation, which leads to confounding the genotype mean performance

with its true value.

Among breeding priorities, stability of performance may be as important as high yield potential. Therefore,

‘genotype x environment’ interactions are of major importance, because they provide information about the effect of

different environments on cultivar performance and have a key role for assessment of performance stability of the

breeding materials.

Developing a wheat cultivar generally results from the selection of valuable recombinants found in manageable

hybrid populations. During the breeding process, they will be grown in a limited set of environments. Evaluation of

breeding material in a wide range of environments seldom is possible, not to mention the multiple environments encoun-

tered by new cultivars released for commercial production. Testing over as wide a range of environments generally is

essential if widely adapted cultivars are to be identified.

Environments are seldom, if ever, duplicated. Variation in an environment at a single location over years can be

as great as those between locations in one year. Therefore, variations in the ‘genotype x environment’ interaction that are

pertinent to wheat breeding problems are those associated with ‘cultivar x year’, ‘cultivar x location’, and ‘cultivar x

year x location’.

We have discussed stability and adaptation of winter wheat. Yield stability has been defined as the ability of a

cultivar to produce an expected yield at the level of productivity of a certain environment (i.e., the cultivar that has no

‘genotype x environment’ interactions). In practice, the wide variation in yield stability are related to the range in

adaptation and response to production inputs. Therefore, wheat cultivars must have sufficient potential to maintain

competitive yields in various environments and react favorably to conditions or increased production inputs.

That practical wheat breeding can make increases in genetic yield potential without substantial loss in yield

stability and adaptation is of question. Some believe that yield potential and yield stability are more or less independent.

Others say that yield stability is inversely proportional to the sum of squares for the ‘genotype x environment’ interaction

attributable to that cultivar. The fact that one cultivar has significantly superior mean yields than another over a wide

range of environments denotes genetic differences in the behavior of different genotypes. However, high mean yield

alone is not necessarily indicative of high stability and wide adaptation.

Finlay and Wilkinson (1963) pointed out that the desired genotype is the one that produces a high mean yield

over a range of environments and has average yield stability in comparison with other genotypes in the same conditions.

They suggested that each nursery mean yield can be considered as a measure of an environment and, thus, an array of

low- to high-yielding environments becomes available from a given set of ecological trials. The response of a particular

cultivar to this range of environments can be estimated by the regression of yield of each cultivar on the mean yield of

the nursery. The regression coefficient (b) is considered as a parameter of yield stability. So, b = 1 denotes cultivars

with average stability; b > 1 are less stable cultivars, and b < 1 denotes very stable cultivars.

Eberhart and Russel (1966) developed this concept of stability and suggested the use of two stability parameters

when describing the performance of one cultivar over a range of environments. They proposed that the regression of

each cultivar on an environmental index and a function of the squared deviations from regression would provide more

useful estimates of yield stability parameters. The environmental index is a coded deviation of each environment from

the grand mean over a given range of environments. Environmental index is obtained for each environment by subtract-

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

ing the grand mean of all cultivars over all environments from the mean of all cultivars in each environment. This forces

the regression of the mean of all cultivars on the environmental index to have unit slope (b = 1). Therefore, a stable

cultivar can be defined as one that have above average performance in all environments, a unit regression coefficient (b =

1), and a deviation from regression as small as possible (Sd2 = 0).

Evaluating yield stability and adaptation of the ARDS Turda winter wheat cultivars.

We evaluated the data from 22 ecological yield trials to examine the contributions of the ARDS Turda winter wheats to

increases in yield and stability of performance in wheat production. The trials are from seven locations and three years

(1998–2000) plus one location with one year (1998); a total of 22 trials. These locations are representative of the diverse

environmental conditions in Romania. Experimental cultivars in each trial did not exceed 25, including the long-term

check Bezostaia 1, used for comparison to newer cultivars and nursery performance over the years. The trials usually

were evaluated in a RCB with six replications. Previous crop, seeding date, and fertilization were different at each

location and conformed to local practices. Because part of cultivars in the nursery are changed annually and may

influence stability parameters, we chose only 11 cultivars that remained in all trials over the experimental years. The 22

trials included 10 Romanian winter wheats plus Bezostaia 1, the check considered to have had a fairly stable yield and

satisfactory adaptation. Five of the 10 Romanian cultivars analyzed were released by the ARDS Turda wheat-breeding

program.

‘Cultivar x year’, ‘cultivar x location’, and ‘cultivar x year x location’ interactions were significant, indicating

that the yield performance of the cultivars varied with the environments.

Stability parameters, computed according to the Eberhart and Russel model, were used to describe the perfor-

mance of cultivars over environments. According to the model, the environmental index as an independent variable (x)

was obtained for each of 22 environments as the mean of those 11 cultivars minus the grand mean (mean of the 11

cultivars in all 22 environments). The mean yield of each cultivar in each environment (y) was than regressed upon the

environmental index. The statistical mean yield, regression coefficient (b), and coefficient of determination (r2) are

currently used to evaluate the stability of yield over environments. We prefer coefficient of determination instead of

deviation from regression because it directly gives predictability of a cultivar in relation to the environmental index.

Although the deviation from regression must be as small as possible (approaching 0), the desired coefficient of determi-

nation is one that approaching 1 when considerable confidence can be attributed on one environment’s measurement of a

cultivar’s performance and adaptation.

Stability parameters for the yield of

the ARDS Turda winter wheats in the 22

ecological trials and the check Bezostaia 1

are presented in Table 1. According to the

statistical model, the mean yields correspond

to an environmental index value of 0.

Directly evaluating the percentage gain in

yield attributed to cultivar improvement is

relative to Bezostaia 1. In our case, this

value was between 8 % for Transilvania and

17 % for Ariesan. At the same time, in

comparison with Bezostaia 1, our cultivars

have had regression coefficient of 1 or

slightly higher, except for Turda 95, which

has a lower value. In addition, coefficient of

determination values equal to or higher than

Bezostaia 1 show that the cultivar response

to environments is predictable to a considerable degree. Turda 95, which has a lower slope of regression (b = 0.87),

seems to be well adapted to suboptimal environmental conditions.

A higher regression coefficient is desirable for high-yielding cultivars because they must be responsive to

favorable conditions or increased cultural input. Above average performance in all types of environments must be

Table 1. Stability parameters for yield of five ARDS Turda winter

wheat cultivars compared with long-term check Bezostaia 1 and

grown in 22 yield trials in northcentral Romania (1998–2000).

Percent mean yield is expressed as a percent of Besostaia 1.

Mean yield Regression Coefficient of

coefficient determination

Cultivar q/ha % (b) (r2)

Transilvania 54.4 108 1.00 0.91

Ariesan 58.7 117 1.07 0.93

Apullum 57.9 115 1.11 0.90

Turda 95 58.2 116 0.87 0.86

Turda 2000 58.2 116 1.07 0.86

Bezostaia 1 50.3 100 0.96 0.86

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

maintained. The regression of the yield of

Transilvania (released in 1982) and Turda

2000 (released in 2000) on environmental

indexes compared with the Bezostaia 1 check

are shown in Fig. 1. Differences in the mean

yield of Transilvania and Turda 2000 relative

to Bezostaia 1 demonstrate a continuous yield

advance achieved by our wheat-breeding

program during the last 30 years. The

regression lines of the two cultivars are nearly

parallel with that of Bezostaia 1, indicating

that their superiority is maintained across a

wide range of environments. The slope of

Bezostaia 1 is b = 0.96, whereas the slope of

Transilvania is b = 1 and Turda 2000 is b =

1.07. These cultivars tend to be slightly more

favorable to environments. For these two

cultivars, breeding progress to improve yield

potential was accompanied with improved

stability of performance.

The three other cultivars from our

program, Ariesan, Apullum, and Turda 95, had

different regressions of yield on environmental

indices in the same set of trials; graphically

illustrated in Fig. 2. Ariesan, with the largest

mean yield and a reasonable regression

coefficient (b = 1.07), has the highest coeffi-

cient of determination (0.93) denoting a

strong, predictable response to changes in

environmental conditions. The combination

of increased yield potential with good stability

of performance may explain the wide accep-

tance and popularity of Turda-developed wheats like Ariesan. Turda 95, with a larger mean yield (approaching Ariesan)

but low regression coefficient (b = 0.87), sharply contrasts with the smaller mean yield and high regression coefficient of

Apullum. The coefficients of determination were nearly similar for the two cultivars. The larger mean yield of Turda 95

clearly is associated with its higher yield in the poorer environments, whereas Apullum with a larger regression coeffi-

cient seems to be well adapted in favorable environments. The stability parameters in this study do permit comparisons

among cultivars for average yields, stability of performance as a degree of response to changing environments, and the

predictability of response to specified environments. Such comparisons would be useful for judging the release of

cultivars and making recommendations for suitable production conditions and areas of adaptation for different cultivars.

Conclusions and remarks. The final objective of a winter wheat-breeding program is the release of cultivars combining

high yield potential and quality with stability of performance and adaptation. Breeding for resistance to diseases and

different others biotic and climatic stress promote such stability. The high level of winter hardiness of wheat cultivars is

a major requirement for many winter wheat regions. Many genes condition winter hardiness, but the adaptability and

stability represent more complex breeding characters that are controlled genetically and encompass a large number of

known and unknown morphological, physiological, and biochemical attributes. Therefore, breeding for adaptation must

begin with choosing parents for the crosses. They must be well-adapted genotypes that will give valuable hybrid

combinations for the desired cultivar. During the generations of selection, the breeding material needs to be grown in the

different biotic and climatic conditions with which they will interact to allow the breeder to make sound judgements of

chosen material. In addition, testing breeding material in different simulated conditions such as with pathogen inocula-

tions, aluminum toxicity solutions, and sprouting in a mist cabinet, can help achieve the elements of cultivar adaptation.

New techniques of selection or manipulation of genetic material also can aid in developing high-yielding and stable

cultivars. Previously, we suggested that the pedigree selection method, with only a few reselections, may conserve some

heterogeneity in cultivars and buffer against environmental changes resulting in a good stability of performance (Ann

Fig. 2. Regression of the yields of Ariesan, Apullum, and Turda 95

versus the Besostaia 1 (dashed line) check on environmental indicies

in 22 ecological trials.

Fig. 1. Regression of the yield of Transilvania and Turda 2000

versus the Besostaia 1 check on environmental indicies in 22

ecological trials.

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Wheat Newslet 48:113-115). However, we do not exclude the possibility that homozygous genotypes, like pure lines

obtained by double-haploid techniques or other methods, may buffer as any other type of population if selection for

increased stability is applied.

Breeders agree that testing over a wide range of environments is essential if stable and widely adapted cultivars

are to be identified. However, the extensive trial data required for identification stable cultivars becomes available only

in advanced generations, when a cultivar is close to or may be already released. Therefore, the methods for evaluating

yield stability proposed in this study have had a little significant impact in the early generations of selection regarding

breeding wheat for adaptation. Improved evaluation techniques, applied in early generations, should assist in the early

identification of those lines having high yield potential associated with good adaptation in highly variable environments

or in alerting breeders to possible deficiencies in adaptation for other lines.

Based on our results presented here, trends in cultivar response to environments in regional performance

nurseries indicate that breeders must carefully consider the trade-off between maximum yield potential, stability of

performance, and ranges in adaptation during cultivar evaluation. However, convincing breeders to sacrifice high yield

for increased stability and wide adaptation is difficult. Nevertheless, assessing the ‘genotype x environment’ interaction

as a factor in determining the yield potential in the different production conditions will remain the most important tool in

the breeding wheat for yield.

References.

Eberhart SA and Russel WA. 1966. Stability parameters for comparing varieties. Crop Sci 6:36-40.

Finlay KV and Wilkinson GN. 1963. The analysis of adaptation in a plant-breeding programme. Aust J Inst Agric Res

14:742-754.

Bread-making quality research.

Mss. Rozalia Kadar completed her Ph.D. dissertation in December 2002 under the direction of Prof. Dr. Leon S.

Muntean at the University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca. The title of Rozalia’s thesis

was ‘Study of the genotype-environment interaction in achieving of bread-making quality in winter wheat’. Her thesis

research underscores the fact that most wheat quality characteristics are heritable traits and more or less influenced by

environmental conditions and production inputs. The implications of ‘genotype x environment’ interactions in develop-

ment of winter wheat cultivars with improved bread-making quality are discussed.

ITEMS FROM THE RUSSIAN FEDERATION

AGRICULTURAL RESEARCH INSTITUTE OF THE CENTRAL REGION OF NON-

CHENOZEM ZONE

143026, Nemchinovka-1, Moscow region, Russian Federation.

Genetic linkage between endosperm color and caryopsis size in soft wheat hybrids.

V.G. Kyzlasov.

A method for creating a xenia caryopsis color and its inheritance in soft wheat hybrids has been described previously

(Kyzlasov 1998, 2000, 2001). Plants with caryopses of various colors were detected among the progeny with

instaminate flowers. These plants arose through the pollination of a spring soft wheat with pollen from spring barley.

Instead of stamens, this genotype had formed pistils. The segregation of caryopsis color in the F1 hybrid plants was 7

light-colored : 9 pigmented (1.284 light : 1.662 dark). The pigment was produced in the caryopses as the result of

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

complementary interaction of two hypostatic genes determining xenias. In the reciprocal cross, the hybrids did not differ

in their segregation pattern (596 light-colored : 819 pigmented ~ 688 light-colored : 843 pigmented).

In the second and subsequent generations of the hybrid populations, the unpigmented caryopses produced light-

colored grain progeny only. The progeny of plants with pigmented caryopses segregated 7 light : 9 dark, 1 light : 3 dark

or all plants produced pigmented caryopses. Both endosperm and pericarp were colored in the pigmented caryopses.

The genes for caryopsis xenia color have an effect on the color of the forming endosperm immediately after fertilization.

This phenomenon is known in maize, pea, barley, and rye.

Our investigation revealed that grain xenias can be mani-

fested in other features, e.g., caryopsis size. The dark caryopses

obtained from crosses between a light-grained line (female) and a

dark-grained line (male) were significantly bigger than those from

hybrids of dark-grained line (female) and a light-grained line (male).

The ratio the 1,000-kernel weight produced by the dark-grained

hybrid to that produced by the light-grained line was (38.8 : 34.8) x

100 = 111.4 % (Table 1).

The use of grain xenia-color genes makes it possible to mark

and select caryopses within a separate spike that are carriers of the

genes determining large grains. Selecting caryopses of different

colors within separate spikes of F1 hybrid plants indicates that dark-

colored caryopses are signifi-

cantly larger than light-colored

caryopses (see Table 2).

The ratio of the weight

of dark to light caryopses was

(37.1 : 33.0) x 100 = 112.4 % on

average. Linkage between grain

color and size was established in

other experiments. For example,

sweet corn (Zea mays

saccharata) and garden pea

(Pisum sativum) also demonstrate linked inheritance of grain size and variety features.

Dark- and light-colored grains taken from the same spike did not differ in their levels of raw protein, K2O, P2O5,

and gliadin proteins. Glutenins cause dark-grained wheat. The dark-grained wheat obtained in our experiments is

recommended for use when studying the inheritance of grain size in hybrids. The wide distribution of grains of different

colors within the same spike indicates that the difference in the size is exclusively a function of genetic factors. The

environmental influences are identical for all the grains of a spike. By backcrossing coarse-grain lines can be created

that will be analogous to the commercial cultivars with the dark-colored endosperm and pericarp.

References.

Kyzlasov VG. 1998. Wheat grain xenia. In: Proc Conf The theoretical and the practical problems of the genetic,

selection and seed-growing of the grain crop. Moscow region, Nemchinovka P. 43.

Kyzlasov VG. 2000. Grain xenia and its heredity by soft wheat hybrids. In: 2nd Cong Vavilov Soc Genet Select, St.

Petersburg. 1:111.

Kyzlasov VG. 2001. Genes controlling xenia development of the caryopsis in soft wheat. Ann Wheat Newslet 47:142.

Table 1. 1,000-kernel weight (g) of different

light- and dark-grained wheat lines and their

F1 hybrids. Minimum significant difference

(P = 0.05) = 2.5 g

Parental lines

Light-grained 38.3

Dark-grained 39.2

F1 hybrids

Light-grained / light-grained 34.8

Light-grained / dark-grained 38.8

Table 2. Range in weight of caryopsis after selection according to endosperm

color. Values for 1,000-kernel weight are in grams.

1,000-kernel weight

Endosperm Average

color 20 25 30 35 40 45 50 1,000-kernel weight

Light 2 13 33 30 19 3 — 33.0 ± 0.6

Dark — 2 16 36 31 14 1 37.1 ± 0.6

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

AGRICULTURAL RESEARCH INSTITUTE FOR THE SOUTH-EAST REGIONS

Department of Genetics, 7 Toulaikov St., Saratov, 410020, Russian Federation.

The efficacy of the Ut gene from Saratovskaya

57 (Triticum durum subsp. durum) against loose

smut in bread wheat.

A.E. Druzhin.

During two seasons (2001–02), a number of spring bread

wheat lines containing chromatin from T. durum subsp.

durum were studied for resistance to loose smut after

artificial infection. We selected lines with a high level of

resistance to the pathogen. We detected resistance to loose

smut in lines with the spring durum wheat cultivar

Saratovskaya 57 in their pedigree (see Table 1).

Evaluating resistance to loose smut, bunt, and

ergot in spring bread wheat lines containing alien translocations.

A.V. Borozdina.

The Saratov-bred spring bread wheat cultivars and lines containing alien translocations were evaluated under natural

infection by pathogen populations of loose smut, bunt, and ergot. Spring bread wheat lines derived from crossings with

S. cereale; T. turgidum subspp. durum, persicum, and dicoccum; Ag. elongatum; Ag intermedium; and lines with translo-

cations from these species were most susceptible to ergot. The degree of a susceptibility in the lines containing genes

from T. turgidum subsp. durum + Ag. elongatum + S. cereale was higher than lines in lacking rye in their pedigree.

Resistance to loose smut was found in lines with translocations T. turgidum subsp. durum + T. turgidum subsp.

dicoccum + Ag. elongatum (L 836-00), T. turgidum subsp. durum + Ag. elongatum (L 2040 and L 164), T. tudgidum

subsp. durum + Ag. elongatum + Ag. intermedium (L 810-94), T. turgidum subsp. durum + T. turgidum subsp. persicum

(L 589-94), and T. turgidum subsp. durum +Ag. elongatum + S. cereale (L 894-94 and L 255-93).

The study of these lines for resistance to bunt shows that the greatest number of susceptible genotypes in the

groups with translocations from Ag. elongatum, T. turgidum subsp. durum, and their combinations (T. turgidum subsp.

durum + Ag. elongatum) and (T. turgidum subsp. durum + Ag. elongatum + Ag. intermedium). Lines combining resis-

tance to all three diseases were very rare and observed in lines containing T. turgidum subsp. durum + T. turgidum subsp.

persicum line L 589-94.

Genetic control for resistance to leaf rust in spring bread wheat lines derived from crosses with

tetraploid AB-genome species.

S.N. Sibikeev, S.A. Voronina, and V.A. Krupnov.

In the Department of Genetics at ARISER, spring bread wheat lines resistant to leaf rust were obtained from crosses with

T. turgidum subspp. durum, dicoccum, and dicoccoides. These lines were produced by the backcross method. In these

lines, the following ITs to leaf rust were observed: L164 (pedigree: L504/S57//L504, S57 is a spring durum wheat) – IT

= 2–2; L196 (pedigree: S58/T. turgidum subsp. dicoccum*3//S58) – IT = 0;–1; L2870 (pedigree: S55/T. turgidum subsp.

dicoccoides*3//S55) – IT = 0;. Genetic analyses indicated that the resistance in L164 was determined by two recessive

genes, in L196 by two dominant genes, and in L2870 by one dominant gene. Allelism tests detected that these Lr genes

are different from Lr14a and Lr23 and from each other.

Table 1. Reaction of cultivars and lines of spring bread

wheat to race 23 of loose smut during 2001 and 2002.

% of plants sporulating

Cultivar/line 2001 2002

L503 58.3 59.5

L504 64.2 59.3

L222 67.5 59.4

Saratovskaya 58 (S58) 70.3 65.2

L504/S57*2//L504/3/S58 5.8 3.1

L503/S57//L503 5.3 4.7

L503/S57//L503/3/L222 0.0 0.0

Saratovskaya 57 (S57) 0.0 0.0

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

The evaluation of spring bread wheat cultivars for resistance to stripe rust in 2002.

S.N. Sibikeev and A.E. Druzhin.

Stripe rust of bread wheat in the Saratov district of the Volga Region of Russia occurs seldom and the severity of the

epidemics is usually weak. Nevertheless, in the southwestern part of the Saratov district, epidemics of this disease were

observed during the last 2 years. A major part of this area is sown with spring bread wheat cultivars L503, L505,

Belyanka, and Dobrynya. L503, L505, and Dobrynya had an IT of 0 and that of Belyanka was a 3. L503, L505, and

Dobrynaya have resistance gene Lr19, and Belyanka has Lr23+Lr14a. Resistance to stripe rust in L503, L505, and

Dobrynya was surprising, because there is no data regarding the resistance of Agropyron translocation with Lr19 to P.

striiformis tritici.

The reaction of the bread wheat cultivars and lines to loose smut and bunt.

A.Yu. Buyenkov, A.E. Druzhin, V.A. Krupnov, Yu.V. Lobachev, and M.R. Abdryaev.

We compared the resistance of bread wheat cultivars and lines to loose smut and bunt in an artificial infection. Fourteen

cultivars and lines bred at ARISER were infected with spores of loose smut and bunt. The initial inoculum of bunt was

collected from the susceptible line L894. Two

pathotypes of loose smut were collected from

cultivars L505 and Saratovskaya 60, which are

susceptible to the named races L 505 and S 60,

respectively.

The reactions of bread wheats and lines

to bunt and loose smut after artificial inoculation

are given in Table 2. Lutescens 62 is resistant to

bunt but is moderately susceptible to race L 505 of

loose smut and highly susceptible to S 60. L235-

01, the line most susceptible to bunt, was resistant

to race L 505 and moderately susceptible to S 60.

L 2040 was resistant to both races of loose smut

but moderately susceptible to bunt (36 %). The

majority of bread wheat lines were more suscep-

tible to bunt than resistant to loose smut (Albidum

188, L502-01, L105, and L 108). In most cases,

the cultivars and lines were more susceptible

when inoculated with race S 60 of loose smut.

Approximately similar degrees of susceptibility to

bunt and race S 60 of loose smut were observed in

L780 and between bunt and races L 505 in L400,

L154, and L199. A correlation was detected

between the percent susceptible to loose smut and

bunt.

The distribution of Puccinia triticina pustules on the flag leaves of Saratov soft spring wheat

cultivars.

O.V. Subkova.

In practice, we are selecting soft spring wheat cultivars for general resistance to leaf rust. Quantative estimations, both in

the field and greenhouse, include four signs that in total are considered as the final expression of a given interaction.

Most methods for estimating pustule distribution contain a number of errors. A wider selection of characteristics is

Table 2. Percent infection of bread wheat cultivars and lines to

bunt and loose smut pathotypes under conditions of artificial

infection in 2002.

% of plants sporulating

Loose smut

Cultivar/line Bunt L 505 S 60

Lutescens 62 8.0 47.0 71.0

L503 19.0 29.0 47.0

Yuogo-vostotchnaya 2 22.0 34.0 48.0

Albidum 188 23.0 45.0 48.0

L400 25.0 29.0 59.0

L181 rq 30.0 47.0 51.0

L502-01 33.0 49.0 77.0

L154 Rq 35.0 35.0 55.0

L2040 36.0 4.0 5.0

L780-01 44.0 11.0 47.0

L199-01 46.0 50.0 38.0

L105-01 61.0 19.0 13.0

L108-01 62.0 22.0 36.0

L235-01 68.0 9.0 36.0

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

necessary to increase the accuracy. Finding a new phenotypes (up to two) to help determine the various estimations is

desirable.

We visually observed the pustule arrangement on the flag leaves of 12 Saratov soft spring wheat cultivars of the

Ugo-Vostok Scientific Research Institute in the 2000–01 season by sketching. The differences between cultivars were

assessed from pictures of pustule distribution along the leaf blade. Our hypothesis about pustule distribution includes

several points:

in general, in each of the 12 Saratov cultivars, we could determine a typical arrangement from the background

pustule arrangement of the flag leaves and defined this as the abstract picture of pustule arrangement

(aPDP);

each aPDP of a cultivar consists of a definite design type (from 1 up to 9), one of which can be seen only in

single cultivar; others in several; and

a formula describing the pustule arrangement (FDDP) for the resistant and susceptible Saratov wheat cultivars

FDDP includes three traits; the number of dominant designs at the bottom, in the middle, and at the tip of the

flag leaf.

These data are summarized and expressed by the symbol G, with the index n (1–10). These differences in aPDP

are not accidental and are connected with the peculiarities of genotype, plant habit, and the external influences during

epidemic pathogen development.

AGRICULTURAL RESEARCH INSTITUTE FOR THE SOUTH-EAST REGIONS

Department of Genetics, 7 Toulaikov St., Saratov, 410020, Russian Federation.

ALL RUSSIA RESEARCH INSTITUTE OF PHYTOPATHOLOGY

143053, Bol´shie Vyazemy, Moscow region, Russian Federation.

Determining the genotypes of resistant wheat lines using test pathotypes of the Puccinia triticina.

I.F. Lapochkina and E.D. Kovalenko, and A.I. Zhemchuzhina, T.M. Kolomietz, and D.A. Solomatin (Institute of Phyto-

pathology).

Breeding for resistance to the rust fungus is a great problem for many wheat-cultivating countries such as the Russian

Federation, the U.S.A., Canada, Argentina, Brazil, and Australia. One way to solve this problem is to screen for resis-

tance in the world’s wheat germ plasm. After testing common, spring-wheat cultivars grown in the territory of the

former Soviet Union, only 8–9 leaf rust-resistance genes were identified (Singh et al. 1995). The genetic diversity of

winter wheat cultivars is not large; 95 % of the cultivars included in the State Register of Russian Federation in 1998

were the progeny of Bezostaya 1 and Mironovskaya 808 (Martynov and Dobrotvorskaya 2001).

Enriching wheat germ plasm with genes of wild species and establishing new combinations of resistance genes

will increase significantly the efficacy of breeding wheat for immunity. The cytogenetic stock collection created at the

Agriculture Research Institute of Non-Chernozem Zone contains common wheat genotypes with chromosomes added

from Ae. speltoides (over 60 genotypes that are grouped into 16 clusters according to disease resistance and morphologi-

cal traits) (Lapochkina and Volkova 1994; Lapochkina et al. 1998, 2001). The collection also includes hexaploid

genotypes with alien material from Ae. speltoides, Ae. triuncialis, T. kiharae, and S. cereale. Several stable addition lines

of spring wheat obtained by means of wide hybridization with the spring wheat Rodina and the ph1b mutant with Ae.

speltoides and Ae. triuncialis species were used in this research and are described in Table 1.

Lines k-62903 and k-62904 are of the lutescens type. These lines have a long stem (90–100 cm), a lax

multiflowered ear, and anthocyanin-colored anthers. Line k-62905 belongs is a milturun type. This line has a short

stem, lacks wax on the spike, and has short awn-like sprouts on the ear apex. Line 149/00i is characterized by late

ripening, anthocyanin-colored anthers and straw, and waxless spikes. Line 102/00i is T. aestivum subsp. spelta with a

dense spike. This line has anthocyanin-colored anthers and lacks wax. Line 82/00i has a short stem, lax multiflowered

spikes with big glumes, and elongated teeth on the lemma. Line 76/00i is characterized by the presence of the morpho-

100

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Table 2. The genotypes of the Puccinia triticina pathotypes used in to

identify leaf-rust resistance genes in 13 lines with wheat–alien transfers.

191-7 1,3a,3bg,10,11,14a,14b,15,17,18,21,27+31

238-15 1,2a,2b,2c,3a,3bg,3k,10,11,15,17,18,19,20,21,32,36

242-14 1,2c,3a,3bg,3k,10,11,14a,14b,16,17,18,20,21,25,27+31

245-21 2c, 3a,3bg,3k,10,11,14b,16,17,18,21,27+31

249-6 2b,2c,3a,3bg,3k,10,11,14a,14b,16,17,18,21,26,27+31,32,36

261-7 1,2a,2b,2c,3a,3bg,10,11,14a,14b,15,17,18,20,26

277-23 1,2a,2b,2c,3a,3bg,3k,11,14b,17,18,20,21,25,26,27+31

277-14 1,2a, 2b,2c,3a,3bg,10,11,14a,15,17,18,21,26,27+31

262-6 1,3a,3bg,3k,10,11,14a,14b,16,17,18,20,21,23.25

98-3 1,2a,2b,2c,3a,3bg,3k,11,14a,14b,15,17,18,20,21,26.

270-7 3a,3bg,3k,10,11,14b,15,16,17,18,19,20,21,25,27+31

257-3 1,2a,2b,2c,3a,3bg,3k,11,14a,14b,16,17,18,20,21,26,27+31

277-26 1,2a,2b,2c,3a,3bg,11,14a,14b,17,20,21,26.

258-13 1,2a,2b,2c,3a,3bg,3k,10,11,14a,14b,15,17,18,20,21,27+31,28

269-7 1,2c,3a,3bg,3k,10,11,14b,17,18,20,21.

logical features

of wild species;

thin, anthocya-

nin-colored

straw and low

spike density.

Two telomeric

SPELT 1

repeats are

visualized in the

karyotype of

this line after

FISH (Salina et

al. 2000). Line

87/00i has thin

straw and is

susceptible to

powdery

mildew. Line

72/00i is

characterized by

a light-red spike

color, low spike density, and narrow, lancet-shaped spike scales. This line is resistant to powdery mildew (10 % infec-

tion). Lemmas that adhere to the kernel on the side groove and the existence of a long (over 9 cm) spike are typical of

line 99/00i. The line also is susceptible to powdery mildew (40 % infection). Line 85/01i has a low spike density and

anthocyanin-colored anthers and straw. Line 97/01i has thin straw and the lemma adheres to the kernel. Line 132/01i is

an awned form of T. aestivum.

During the last 5 years, all lines

showed a high level of resistance to leaf

rust inoculation (genotype of the popula-

tion 1, 2a, 2b, 2c, 3bg, 3k, 10, 11, 14a,

14b, 16, 17, 18, 20, 21, 23, 25, 26,

27+31, 30) in the field. Fifteen isolates

collected from the natural

uredopopulations of the pathogen in the

Central, Low-Volga, Middle-Volga,

North-Caucasian, and West-Siberian

regions of the Russian Federation were

used as test cultures. Pathotypes of P.

triticina carried from 12 to 18 virulence

genes (Table 2). Disease symptoms were

estimated according to the 5-point scale

of Mains and Jackson (1926). Infection

types 0, 0; 1, 2, and X- mean that a

sample possesses resistance genes

whereas types 3, 4, and %+ indicate their

absence.

As a rule, the investigated lines were resistant to the pathogen penetration with ITs of 0, 0; or 1, and 2. A

susceptible reaction to some pathotypes suggested that lines k-62905, 149/00i, 82/00i, and 102/00i had a combination of

Lr1+Lr10, Lr10+Lr26, Lr10+Lr26, and Lr27+Lr31 genes, respectively. In addition, lines 82/00i and 102/00i each had

one additional, unidentified resistance gene.

Lines k-62903 and k-62904 presumably have new resistance genes from Ae. speltoides. The alien transloca-

tions and substitutions (T2BL·2SL for k-62903, T1BL·1SS and T5AL·5SL for k-62904, and 7A/7S substitution in k-

62905) were identified previously by differential C-banding of chromosomes in k-62903, k-62904 and k-62905

Table 1. Field reaction to powdery mildew and leaf rust and identification of Lr genes in wheat–

Aegilops lines.

% infection in field Assumed

Line Origin mildew leaf rust resistance

k-62903 Rodina/Ae. speltoides 0 0 juvenile gene(s)

k-62904 Rodina/Ae. speltoides 0 0 juvenile gene(s)

k-62905 Rodina/Ae. speltoides 10 20/2 Lr1 + Lr10

149/00i ph1b/Ae. speltoides 00 Lr10 + Lr26

102/00i Rodina/Ae. speltoides (10 kR) 40 0 Lr27 + Lr31 +

82/00i Rodina/Ae. speltoides (10 kR) 0 0 Lr10 + Lr26 +

76/00i Rodina/Ae.speltoides (10 kR) 0 0 adult-plant gene(s)

87/00i Rodina/Ae.speltoides (10 kR) 30 0 juvenile gene(s)

72/00i Rodina/Ae. speltoides (10 kR) 0 0 juvenile gene(s)

99/00i Rodina/Ae. speltoides (10 kR) 5 0 juvenile gene(s)

85/01i Rodina/Ae. speltoides (10 kR) 0 0 juvenile gene(s)

97/01i Rodina/Ae. speltoides (10 kR) 0 0 juvenile gene(s)

132/01i Rodina/Ae. triuncialis (5 kR) 20 0 Lr28 +

101

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

(Lapochkina et al. 1996; Pukhalsky et al. 1999). The presence of resistance genes in these lines probably is related to

these translocations.

When lines 87/00i, 99/00i, 85/01i, and 97/01i were inoculated with leaf rust test pathotypes, they showed only

the resistant-type reaction (0, 0;); suggesting that the resistance genes from Ae. speltoides function in both seedlings and

adult plants. Line 76/00i was susceptible to infection by 11 pathotypes and resistant to four. The presence of APR genes

in this line possibly are related to the presence of Ae. speltoides chromosome 4S in the karyotype. For line 72/00i, the

heterogenic type of reaction (x-) was found in the case of three pathotypes, the 12 remaining pathotypes exhibited a

resistant reaction (0, 0;). We believe that juvenile resistance genes may be present in this line. Lr28 and additional

unidentified resistance genes from Ae. triuncialis were found in line 132/01i.

Conclusions. The testing of 13 wheat–Aegilops lines with leaf rust pathotypes with known genotypes showed

that most lines had juvenile genes of resistance. Line 76/00i with APR genes was identified. All the lines were classified

into three groups: 1) those with unidentified resistance genes from Ae. speltoides (k-62903, k-62904, 72/00i, 85/01i ,

97/01i , 99/00i, 87/00i, 76/00i); 2) those with known genes of resistance (k-62905 and 149/00i); and 3) those with known

resistance genes and an additional unknown resistance gene (82/00i, 102/00i, and 132/01i).

References.

Lapochkina IF. 2001. Genetic diversity of “Arsenal” collection and its use in wheat breeding. In: Abstr Internat Appl

Sci Conf Genetic Resources of Cultural Plants. 13–16 November, 2001, St. Petersburg, Russian Federation. Pp.

133-135.

Lapochkina IF, Solomatin DA, Serezhkina GV, Grishina EE, Vishnyakova KhS, and Pukhalskiy VA. 1996. Common

wheat lines with genetic material from Aegilops speltoides Tausch. Rus J Genet 32(12):1651-1656.

Lapochkina IF and Volkova GA. 1994. Creation of collection of lines spring common wheat, substituted and supple-

mented with chromosomes of Aegilops speltoides Tausch. Rus J Genet 30(suppl):86-87.

Lapochkina IF, Yatchevskaya GL, Kyzlasov VG, Solomatin DA, Vishnyakova KhS, Pogorelova LG, and Lasareva EN.

1998. The production, cytology and practicality of disomic addition lines of T. aestivum-Ae. speltoides. In: Proc 9th

Internat Wheat Genetic Symp (Slinkard AE ed). University Extension Press, Saskatoon, Saskatchewan, Canada.

2:67-69.

Mains EE and Jackson HC. 1926. Physiologic specialization in the leaf rust of wheat, Puccinia tritici Erikss. Phytopath

16(2).

Martynov SP and Dobrotvorskaya TB. 2001. Pedigree-analysis of modern cultivars of winter common wheat. Selektsia

i Semenovodstvo. N 1-2:47-54.

Pukhalski VA, Iordanskaya IV, Badaeva ED, Lapochkina IF, and Bilinskaya EN. 1999. Genetic analysis of the sign

“absence of wax bloom on an ear” of the line of common wheat. Rus J Genet 35(9):1223-1227.

Salina EA, Adonina IG, Efremova TT, Lapochkina IF, and Pshenichnikova TA. 2000. The genome-specific

subtelomeric repeats for study of introgression lines T. aestivum x Ae. speltoides. In: Abstr 11 EWAC Conf, dedi-

cated to the memory of O.I. Maystrenko, 24–28, July, 2000. Pp. 181.

Singh RP, Morgunov A, and Huerta-Espino J. 1995. Genes conferring low seedling reaction to Mexican pathotypes of

Puccnia recondita f. sp. tritici, and adult-plant responses of recent wheat cultivars from the former USSR. Euphytica

81(3):225-234.

INSTITUTE OF COMPLEX ANALYSIS OF REGIONAL PROBLEMS

Karl Marx str., 105 A, kv. 167, Khabarovsk, 680009, Russian Federation.

Technological characteristics of spring wheat cultivars developed in the far-eastern Russian

Federation.

Ivan M. Shindin, Elizoveta N. Meshkova, and Olga V. Lokteva.

The grain market in the far-eastern part of the Russian Federation is mainly imports from abroad and the central region

of the country. The cost is high. Because the far-eastern region has sufficient land and a favorable environment for the

production of spring wheat, the area is completely capable of providing the population of the region with bread and

102

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

bakery products. To solve this important problem, increasing the wheat yield from 1.0–1.2 t/ha to 1.8–2.0 t/ha, extending

the area under cultivation from 200,000 to 450,000–500,000 ha, and having high-quality cultivars is necessary.

In the Russian

Federation, T. aestivum

cultivars are classified

into five groups

according to their grain-

technological character-

istics into the categories

strong, valuable,

medium quality (filler),

satisfactory, and weak

(Table 1).

The term

strong means wheat

with high-quality

protein content that

forms a dough good for

intensive mixing and

long fermentation,

provides for a high

volume of bread, and

has good mixing quality.

The mixing quality is

understood to be the

capability of strong

wheat flour to improve

baking quality of a weak

wheat flour. The higher

the mixing quality of the

flour, the less quantity

of flour is required as a

component of a mixture

(from 50–20 %).

Valuable and medium

(filler) wheats make

high-quality bread, but

they do not improve the

baking quality of weak

cultivars. Flour from

weak wheat when not

combined with a strong

wheat flour is not good

for bread baking.

Wheat quality

problems are of great economic importance. If 100 g of high baking-quality wheat yields 115 kg of bread, then a low

baking-quality wheat will yield only 9l kg (Pumpyansky and Semyonov 1969). Thus, the main importance for obtaining

high-quality bread depends on the cultivar.

The far-eastern region has the proper cultivar resources (Shindin 1996; Shindin and Bochkaryov 2001). In

2002, 13 cultivars of soft spring wheat were released for cultivation; 11 are from breeders in the far east (Amurskaya 75,

Amurskaya 1495, Amurskaya 90, Dalnevostochnaya 10, Zaryanka, Lyra 98, Monakinka, Primorskaya 14, Primorskaya

21, Primorskaya 39, and Kabarovchanka) and two are from other regions (Krasnofimskaya 90 and Priokskaya). Al-

Table 1. Classification of Triticum aestivum cultivars according to bread-making quality.

Good

Quality indicators Strong Valuable filler Satisfactory Weak

grain hardness hard and medium hard — — —

vitreousness, %

(not less than) 60 50 50 40 —

protein content in grain

(not less than) 14 13 12 11 8

gluten content in grain, %

(not less than) 2 ~25 24 22 15

gluten content in

70 % flour output, %

(not less than) 32 29 27 25 20

dough dilution,

pharinograph units,

(not more than 30–60 80 120 150 > 150

valorimetric number

farinograph units,

(not less than) 70–85 55 45 00 < 80

dough deformation,

alveograph units,

(not less than) 280 260 240 180 < 180

dough elasticity

(alveograph), mm

(not less than) 80 70 60 50 < 50

bread output from

100 g of flour, ml

(not less than) 1,200 1,100 900 800 < 800

baking quality mark

(not less than) 4.5 4.0 3.5 3.0 < 3.0

103

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

though no strong wheat cultivar is among these wheats, most are good fillers and are of good quality according to their

technological evaluation. All have good agronomic characteristics (high grain yield and resistance to lodging, disease,

sprouting, and shattering). The technological and agronomic characteristics of eight far-eastern cultivars follow.

Amurskaya 5. This cultivar was bred at the former Amur Agroexperimental Station (now the Russian Soybean

Research Institute, Blagoveshensk), which is situated in the Amur region, a main wheat granary in the far-eastern

Russian Federation. The grain is of average size and vitreous (60–78 %). The 1,000-kernel weight is 27–34 g. The 1 L

weight is 750–760 g. Baking quality is good. Amurskaya 5 belongs to the valuable class of wheat cultivars. Grain

protein content is 14.1–17.8 %, gluten content is 27–40 %, and flour strength is 280–411 units as measured by

alveograph. The bread output from l00 g of flour is 620–1,150 ml. Baking quality is 3–4.5. The cultivar is resistant or

moderately resistant to lodging, shattering, and P. graminis. Grain yield is 2–2.5 t/ha.

Amurskayag 90. This cultivar was bred at the Ear Eastern State Agricultura1 University, Blagoveshensk. The

grain is egg-shaped, red, and vitreous with a shallow groove. The 1,000-kernel weight is 32–35 g. According to

technological evaluations, Amurskayag 90 is a satisfactory filler, threshes well, and is resistant to U. tritici and P.

triticina but susceptible to S. nodorum and P. gramminis. Potential yield is 4–4.5 t/ha, with an average yield of 2.5 t/ha.

Dalnevostochnaya 10. This wheat was bred at the Far Eastern Research Institute of Agriculture, Khabarovsk.

The egg-shaped grain is red. The 1,000-kernel weight is 30–38 g. Bread-making quality is medium to good and the

cultivar is a satisfactory filler. Grain vitreousness is 65 %, and protein content is 28.5–37 %. Flour strength is 230–360

units as measured by alveograph and valorimetric number is 50 units as measured by farinograph. The bread output

from 100 g of flour is 650–1,050 ml. Bread-making quality is 2.7–4.1. The cultivar is resistant to lodging and moder-

ately resistant to P. triticina and P. graminis. Commercial yield is 2–2.5 t/ha with a potential yield of 5 t/ha.

Zaryanka. Bred at the Ear Eastern Research Institute of Agriculture, Khabarovsk, Zaryanka has a grain-protein

content of 14–16.7 %, a vitreousness of 60–65 %, gluten of the first-class quality at 28–30 %, and a flour strength of

320–380 units as measured by alveograph. The bread output from l00 g of flour is 960–1,060 ml. The cultivar belongs

to the valuable class. Zaryanka is more resistant to U. tritici, F. graminearum, shattering, and sprouting as compared

with the standard and yields between 2.5–3 t/ha.

Lyra 98. Lyra 98 was bred at the Far Eastern Research Institute of Agriculture, Khabarovsk and released for

growing in the far-eastern region in 2002. Grain protein content is 16–l7 %, vitreousness 60–70 %, gluten content is 30–

38 %, and flour strength is 450–520 units as measured by alveograph. The bread output from l00 g of flour is 1,100–

1,200 ml. This cultivar is resistant to lodging, sprouting, and U. tritici and moderately resistant to F. grameniarum.

Potential yield is 4.6–5.0 t/ha.

Primorskaya 14. Bred at the Primorskey Research Institute of Agriculture, Ussuryisk, this cultivar has red, egg-

shaped grains with a medium groove and of small to average size. The 1,000-kernel weight is 30–36 g. Baking quality

is from satisfactory to good. Vitreousness is 50–78 %, grain protein content is 15–16.8 %, flour gluten is 34.6–39.8 %,

and flour strength is 255–343 units as measured by alveograph. The bread output is 620–1,020 ml with a bread-making

evaluation of 2.6–3.8 marks. Primorskaya 14 is resistant to lodging except in rainy years, resistant to P. graminis, and U.

tritici and moderately susceptible in rainy years to P. triticina and F. graminearum. The commercial yield of

Primorksaya 14 is 2.5 t/ha with a potential yield of 5 t/ha.

Primorskaya 21. This cultivar was bred at the Primorskey Research Institute of Agriculture, Ussuriysk. The

grain is red and oval with a small, narrow groove. The 1,000-kernel weight is 30–42 g. This wheat is of satisfactory

baking quality and a good filler. Grain protein content is 14.7–17.6 %, flour gluten content is 37 %, and flour strength is

270–320 units as measure by alveograph. The bread output from 100 g of flour is 800–1,030 ml. Baking evaluation is

3.6–4 marks. Primorskaya 21 is resistant to lodging and moderately susceptible to P. triticina and F. graminearum.

Average yield is 2.5–3 t/ha with a potential yield of 5 t/ha.

Primorskaya 39. The cultivar was bred at the Primorskey Research Institute of Agriculture, Ussuriysk. The

grain is red, rounded, and vitreous with a medium groove. The 1,000-kernel weight is 30–34g. Baking quality is good to

excellent. Grain protein content is 13–15.9 %, gluten content is 33 %, and flour strength is 440 units as measured by

alveograph. The bread output from l00 g of flour is 810 ml. Baking evaluation is 4.6 marks. Primorskaya 39 is resistant

104

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

to lodging and moderately susceptible to P. triticina and F. graminearum. The average yield is 3.5 t/ha with a potential

yield between 4.5 and 5 t/ha.

Khabarovchanka. This cultivar was bred at the Far Eastern Research Institute of Agriculture, Khabarovsk. The

large, red, egg-shaped grains have a narrow, medium groove. The 1,000-kernel weight is 36–45. Grain protein content

is 14–16 %, grain gluten content is 28–31.5 %, and flour gluten is of the first and second quality at 35.7 %, and flour

strength is between 280–350 units as measured by alveograph. The bread output from l00 g of flour is 900–1,000 ml.

Baking evaluation is 3.6–4.5 marks. Khabarovchanka is highly resistant to lodging, U. tritici, P. graminis, and P.

triticina. This cultivar is of the intensive type and has a good responsiveness to improved growing conditions. Commer-

cial yield is 3–4 t/ha with a potential yield of 5 t/ha.

In the future, this list of cultivars will increase as new cultivars with high value and strength are released by

breeders from the far-eastern region.

References.

Pumpyanskey AY and Semyonova LV. 1969. The improvement of technological characters of Triticum aestivum.

Moscow. P. 87 (in Russian).

Shindin IM. 1996. Spring wheat and barley selection in far eastern Russia. Scientific report, Khakarovsk. P. 55 (in

Russian).

Shindin IM and Bochkaryov YY. 2002. Plant and cultivar resources of agricultural crops in far eastern Russia.

Ussurlysk, Biribidjan. P. 193 (in Russian).

OMSK STATE PEDAGOGICAL UNIVERSITY

Chemico-Biological Faculty, nab. Tuchachevskogo, 14, Omsk, 644099, Russian

Federation.

Chromosome number variations caused by the 2,4-dichlorphenoxyacetic acid in durum wheat

calli and regenerant plants cultivated in vitro.

Natalia A. Kuzmina.

Introduction. Totipotence is a property of plant cells that makes inheriting information following alterations in the

environment and causes the regenerating of plants. The genetic properties of cell populations cultivated on different

artificial media and the possibility to introducing hereditable mutations caused by different mutagenic substances are

very important (Butenko 1964). Auxins, used for the transformation of isolated plant cells and tissues in culture, were

capable of modifying some stages of the mitotic cycle (Gamburg et al. 1990). In addition, the callus tissue itself was

shown to be heterogeneous and genetically unstable, being affected by certain environmental compounds such as light,

temperature, and nutrition (Shamina 1970). Cytogenetic analysis of durum wheat callus tissues described the ploidy

level and revealed a certain genotype and cultivation time (Bennici et al. 1988; Morozova 1991; Yurkova 1989; Yurkova

et al. 1985). We studied variations of the chromosome number in durum wheat callus tissue and regenerated plants as

they relate to some components of the nutrient medium.

Material and methods. Mature seeds of the durum wheat cultivar Altayskaya Niva were used to obtain callus tissues.

The upper part of mature embryos were cut and placed on a nutrient-agar medium. Hamburg medium (B5) (Gamborg et

al. 1968) supplemented with 2,4-dichlorphenoxyacetic acid (2,4-D) in concentrations of 2, 4, or 6 mg/l and Murashige-

Skoog medium (MS) (Murashige and Skoog 1962) supplemented with 2,4-D at 4 mg/l were used to induce callus

growth. Each group consisted of 30–35 explants. Cultivation was in ‘100 x 20-mm’ glass tubes with 10 ml of medium

at 24˚C under continuous fluorescent light. After 6–7 weeks, the calli were transferred to another tube containing the

same medium, but without the hormone supplement, to induce organogenesis. Cytogenetic analysis of the callus tissues

and tissues of the apical root-tip meristem was with temporary squash preparations in acetocarmine. The samples were

treated with 0.1 % colchicine at 4˚C and fixed in a solution of alchohol:acetic acid (3:1). The significance of the

variation between treatments was determined using the Student’s t test (Lakin 1990).

105

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Results and discussion. After the explants were placed on the agar medium containing 2,4-D, callus formation was

observed after 6 weeks of cultivation. Transferring the calli to hormone-free medium caused a range of morphogenic

processes such as the continuous proliferation of nondifferentiated cells (nonmorphogenic callus), the appearance of

rhyzogenesis zones, shoot induction, and formation of somatic embryoids, the beginning of regenerant plants. The

combination and proportion of these processes depended on the auxin concentration in the primary medium (Kuzmina

1997).

The normal

chromosome number was

2n = 28 in the regenerant

plants (Table 1). Greco et

al. (1984) described

chromosome numbers and

Bennici et al. (1988)

identified mosaics in durum

wheat plants regenerated

from the mesocotyl callus.

Reduction in chromosome

number depended on 2,4-D

concentration and was

detected in the roots of

rhyzogenic callus. When

the calli were grown on a

medium supplemented with

4.0 mg/l of 2,4-D, few cells

had reduced chromosome

numbers, but the proportion

of such cells significantly

increased with higher concentrations of 2,4-D. The data clearly demonstrated that a high concentration of 2,4-D causes a

reduction in chromosome number, at least in root-tip cells, because cells of the nonmorphogenic calli had reduced

chromosome number independently of the 2.4-D concentration. This reduction might be explained by mitotic damage

and reduction in mitosis rate, particularly due to loss of the final phases. The incapability of the proliferating cells to

undergo mitosis might be associated with physiological hyperactivity of the cells maintained in an unusual experimental

environment in vitro and with their concurrence for specific regulatory proteins (Gamburg et al. 1990; Shamina 1970;

Yurkova et al. 1985).

The B5 and MS media contained different proportions of a nitrogen salts, NH4NO3, KNO3, and (NH4)2SO4.

Reduced nitrogen (NH+4 and glycine) prevailed in the MS medium, whereas oxygenated nitrogen (NO-3) prevailed in the

B5. Because genetic instability might be caused by different components of the nutrition medium (Shamina 1970), we

compared results obtained with two nutrition media of different mineral compounds (B5 and MS) supplemented with

2,4-D in equal concentrations of 4.0 mg/l (Table 2). The regenerated plants retained normal chromosome number (2n =

28) independent of the medium used. Roots of the rhyzogenic calli maintained on the MS were diploid, whereas cells of

the nonmorphogenic calli had a reduced chromosome number. Only cells of calli maintained on B5 contained reduced

chromosome number (2n = 20 or 22). Root-tip cells of the rhyzogenic calli maintained on the B5 contained either

normal or reduced chromosome number (2n = 26). Increasing the reduced nitrogen in the MS probably diminished the

effect of auxin stress and retracted the chromosome reduction. In one sample, nonmorphogenic calli maintained on MS

medium had a single giant cell with 84 chromosomes (not included into the average calculations). Therefore,

myxoploidy of the cell populations maintained in vitro might be the result of either reduction or multiplication the

chromosome number.

One effect of chromosomal instability on the morphogenetic potency of the cultivated cells is difficult to

estimate. Winfield et al. (1995) reported a correlation between the stability of a cell line karyotype and the rate of

regeneration of the progeny plants, whereas others detected an absence of morphogenesis in the stable lines. The

absence of shoots and regenerant plants in our experiments may be associated with a reduced chromosome number in the

cells of nonmorphogenic and rhyzogenic calli. This point needs additional study.

Table 1. Chromosome numbers of the root and callus cells depending on the 2,4-D

concentration in the B5 medium. Numbers are averages with 95 % confidence limits.

Sample origin 2 mg/l 2,4-D 4 mg/l 2,4-D 6 mg/l 2,4-D

Roots of the regenerant plants 28.0 + 0.0 28.0 + 0.0 —

Roots of the rhyzogenic calli 28.0 + 0.0 27.5 + 1.0 26.2 + 1.8

Cells of the non-morphogenic calli 21.2 + 1.0 21.5 + 1.0 20.5 + 1.3

Table 2. Chromosome numbers of the root and callus cells depending on the proportion

of NO-3 and reduced nitrogen in the nutrition media (MS (Murashigi-Skoog) or B5

supplemented with 2,4-D, 4 mg/l). Numbers are averages with 95 % confidence limits.

Sample origin MS B5

Roots of the regenerant plants 28.0 + 0.0 28.0 + 0.0

Roots of the rhyzogenic calli 28.0 + 0.0 27.5 + 1.0

Cells of the non-morphogenic calli 22.7 + 1.8 21.5 + 1.0

106

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

References.

Bennici A, Caffaro L, Dameri RM, Gastaldo P, and Profumo P. 1988. Callus formation and plantlet regeneration from

immature Triticum durum Desf. Embryos. Euphytica 39:255-263.

Butenko RG. 1964. A culture of isolated tissues and physiology of plant morphogenesis. Nauka. 272 p.

Gamborg OL, Miller RA, and Ojima K. 1968. Nutrient requirements of suspension culture of soybean root cells. Exp

Cell Res 50:151-158.

Gamburg KZ, Rekoslavskaya NI, and Shevtsov SG. 1990. Auxin in plant tissues and cells M.: Nauka. 243 pp.

Greco B, Tanzarella OA, and Blanco A. 1984. Plant regeneration from leaf base callus in durum wheat (Triticum durum

Desf.). Cereal Res Commun 12:171-177.

Kuzmina NA. 1997. Dependence of the durum wheat callusogenesis and regeneration on the concentration of 2.4-D in

the nutrition medium. A regulator of plant growth and development. In: Proc IV Internat Conf, Moscow, 24–26

June, 1997, Moscow. Pp. 302-303.

Lakin GF. 1990. Biometrics. M.:Vysshaya shkola. 352 pp.

Morozova SE. 1991. Stability of the regeneratory properties in a wheat calli. Biology of cultvated cells and plant

biotechnology. M.: Nauka. Pp. 256-259.

Murashige T and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures.

Physiol Plant 15:473-497.

Shamina ZB. 1970. Genetics and cytology of tissue culture and a regenerant plants. A culture of isolated organs, tissues

and cells of plant. M.: Nauka. Pp. 129-137.

Winfield MO, Schmitt, Lörz H, Davey MR, and Karp A. 1995. Nonrandom chromosome variation and morphogenic

potential in cell lines of bread wheat (Triticum aestivum L.). Genome 38:869-878.

Yurkova GN. 1989. The current state and the future of an in vitro wheat cultures. Experimental plant genetics in the

improvement of the selection process. Kiev:Naukiva dumka. Pp. 128-139.

Yurkova GN, Levenko BA, and Novozhilov OV. 1985. Ploidy of callus tissue in durum and common wheat. Cytol

Genet 19:264-267.

PRYANISHNIKOV ALL RUSSIAN RESEARCH INSTITUTE OF AGRICULTURE

AND SOIL SCIENCE

Pryanishnikova, 31. Moscow 127550, Russian Federation.

Aluminium tolerance in spring wheat plants at different levels of potassium and low temperature.

N.V. Poukhalskaya and A.I. Gurin.

Aluminum (Al) toxicity is one of the major problems of agriculture worldwide. Some Al-resistant genotypes have been

identified, however, conditions that minimize damage from Al are unclear. Breeding material should possess not only

specific characters but also a set of positive metabolic responses to environmental stresses typical of particular genotype.

On the other hand, we need knowledge of how mineral nutrition and temperature regimes may eliminate the negative

effect of Al toxicity. We investigated the Al tolerance of the spring wheat cultivar L-63/1 at different levels of potassium

and temperatures.

Materials and methods. Seedlings of spring wheat were grown in a water solution in plastic pots (300 ml, 10 plants/

pot). After a 2-day germination (20°C day/18°C night ± 2°C), seedlings were transferred to pots and grown in three

nutrition regimes: (1) H2O + CaSO4 x 10-4 M (control), (2) KCl x 10-3 M + CaSO4 x 10-4 M (low K), and (3) KCl x 10-3 M

+ CaSO4 x 10-4 M (high K). After cooling for 2, 3, and 6 h, the solutions were supplemented with Al+3 (AlCl3, pH = 5.6)

at concentrations of 3 mg/l and 12 mg/l. Two levels of Al+3 were studied in each variant; low (3 mg/l) and high (12 mg/

l). We examined nine variants in total: (1) control; (2) low K; (3) high K; (4) water + low Al level; (5) water + high Al

level; (6) low K + low Al levels; (7) low K + high Al levels; (8) high K + low Al levels; and (9) high K + low Al levels.

The root systems of the plants were dipped in solution during the 10 days. Solutions were replaced every 2 d. On the

sixth day after germination, some of the plants from each treatment were exposed to cold (+8°C) for 2, 3, and 6 h without

light. After a 6-day cold treatment (6 days of reparation), respiration/photosynthesis and linear growth were analyzed in

all plants. The experiment was repeated three times.

107

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Results. Cold treatment showed a positive effect

on root development, increasing the root-system

length (RL) (Table 1). The negative effect of Al+3

was reduced in K+-containing solutions. However,

the RL at low potassium levels was small. There-

fore, the parameter (RL:LL) was the smallest in

variants with low K+.

Minimal RL was formed at low K+. The

ratio of root-system length:seedling length

(RL:SL) also was minimal in this variant. The RL

in plants grown in water essentially decreased (43–

47 %) upon the addition of Al+3 ions to solution

(Table 2).

The addition of Al+3 to the solution

changed respiratory metabolism in the plants. In

the experimental treatment with water, Al+3

decreased respiration intensity by 43-47 % at both

doses. At low K+, respiration intensity decreased

by 20 % at low Al+3 levels and by 34 % at high Al+3

levels. At high K+, the addition of Al+3 to the

solution did not change respiration intensity at 3 mg/ml and decreased the intensity by 39 % at 12 mg/ml. We assume

that the addition of K+ to water solutions reduces Al toxicity. Examination of the reparation period after cold treatment

has shown that the negative effect of Al+3 inhibits growth activation by cold (Table 3). An increase of K+ decreases the

level of Al+3 toxicity

The Al+3 concentration has a greater effect in the presence of K+ in the nutrition medium. The toxic effects of

Al+3 ions by suppressing the

cold reaction of the root

system is manifested by an

increase in length. Opti-

mizing K+ is an essential

prerequisite for decreasing

Al+3 toxicity. In turn, cold

treatment is a factor that is

capable of adaptating plants

to Al+3 toxicity, supported

by our data from respiration

and photosynthesis experi-

ments. In the control plants

grown at room temperature,

Al+3 decreased respiration

and photosynthesis activity

by 30–68 % after day 6

(Table 4). Respiration in

plants kept in the cold

either did not change (in the

presence of K+) or de-

creased by 5–24 %.

Finally, we have shown that

cooling plants significantly

increases the resistance of

respiratory metabolism to

Al+3.

Table 1. The effect of the low temperature on the length of the

root system (cm). LTD05 = 2.43.

Length of cold treatment (h)

K+ level Control 2 3 6

0 48.40 35.4 63.02 53.46

5 x 10-3 M 27.27 35.28 38.96 36.30

5 x 10-2 M 40.92 41.52 62.08 58.38

Table 2. The effect of Al+3 at low and high concentrations on the

root length at different levels of K+ and optimal temperature.

Al level Water Low K High K

0 42.73 ± 1.41 27.80 ± 3.68 43.68 ± 6.60

3 mg/l 22.47 ± 4.20 21.08 ± 4.74 45.18 ± 5.08

12 mg/l 24.41 ± 4.14 18.38 ± 3.82 26.76 ± 4.76

Table 3. The influence of low temperature on the length of root system (cm) at different

K+ levels and Al+3 concentrations.

Cold H2OH

2O+H2O+lowK+lowK+high K+high K+

period (h) (control) low Al+3 high Al+3 low Al+3 high Al+3 low Al+3 high Al+3

0 43.70 22.47 24.41 21.08 18.38 45.18 26.76

2 42.30 26.44 24.02 22.41 21.54 35.59 23.80

3 63.02 23.57 27.10 21.30 17.15 38.10 25.88

6 53.96 22.39 24.28 22.56 20.56 39.38 24.49

Table 4. Variation in respiration and photosynthesis intensity in control plants and after

cold treatment at different K+ and Al+3 levels (% relative to control plants without Al3+).

Cold-treated plants Control plants

Treatment Respiration Photosynthesis Respiration Photosynthesis

H2O+, low Al+3 24 5 44 68

H2O+, high Al+3 18 18 41 47

Low K+, low Al+3 —5 45 57

Low K+, high Al+3 14 10 45 55

High K+, low Al+3 —— — —

High K+, high Al+3 —— 30 42

108

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

SIBERIAN INSTITUTE OF PLANT PHYSIOLOGY AND BIOCHEMISTRY

Siberian Division of the Russian Academy of Sciences, Lermontov str., 132, Irkutsk-33,

P.O Box 1243, Russian Federation, 664033.

The function of different mitochondrial respiratory-chain pathways in winter wheat mitochondria

during short-term cold stress and hardening.

O.I. Grabelnych, S.P. Funderat, T.P. Pobezhimova, A.V. Kolesnichenko, and V.K. Voinikov.

Plant mitochondria have a branched respiratory chain and, in addition to the main cytochrome pathway, have an alterna-

tive pathway that depends on the functioning of alternative cyanide-resistant oxidase (AOX) (Vanlerberghe and McIn-

tosh 1997). Plant mitochondria also are able to oxidize exogenous NAD(P)H because of the presence of additional

NAD(P)H dehydrogenases in their structure (Soole et al. 1990; Soole and Menz 1995; Moller and Rasmusson 1998).

Recently, a number of proteins that effect mitochondrial activity were found and characterized. Among these are plant

uncoupling mitochondrial proteins (plant UCPs) (Ricquier and Bouillaud 2000) and the stress protein CSP 310 (Voinikov

et al. 1998), which cause uncoupling of oxidative phosphorylation in mitochondria. AOX (Takumi et al. 2002), WhUCP

(Murayama and Handa 2000), and CSP 310 (Kolesnichenko et al. 2000) are present in the mitochondria of winter wheat.

Some of these proteins, such as AOX and CSP 310, are induced by cold stress in winter wheat, but others (WhUCP) are

not. Although WhUCP is not induced by cold stress in winter wheat, its homologues in other plant species were shown

to be induced by cold stress (Laloi et al. 1997; Maia et al. 1998; Ito 1999; Nantes et al. 1999). The main functions of

these proteins were established for animals and proposed for plants are thermogenesis, participation in defense against

oxidative stress, and regulation of cell metabolism (Sluse and Jarmuszkiewicz 2002). On the other hand, mechanisms

that control the different electron-transport pathways in mitochondrial respiration under different stress conditions have

not been studied in detail. Using inhibitor analysis that blocks terminal oxidases or respiratory-chain complexes, we

studied the role of individual mitochondrial respiratory chain pathways in total mitochondrial respiration to learn how the

different electron-transport pathways function in cold-resistant, winter wheat mitochondria during short-term cold stress

and hardening.

Materials and methods. Three-day-old etiolated shoots of the winter wheat cultivar Zalarinka were germinated on

moist paper at 26°C. Shoots were cold-stressed at –1°C for 1 h or were hardened at 4°C for 7 days. Mitochondria were

extracted from seedling shoots by differential centrifugation (Pobezhimova et al. 1996). Isolated mitochondria were

resuspended in a medium of 40 mM MOPS-KOH buffer (pH 7.4), 300 mM sucrose, 10 mM KCl, 5 mM EDTA, and 1

mM MgCl2. Mitochondrial activity was recorded polarographically at 27°C using a closed platinum electrode in a 1.4-

ml volume cell (Estabrook 1967). The reaction mixture contained 125 mM KCl, 18 mM KH2PO4, 1 mM MgCl2, and 5

mM EDTA, pH 7.4. 10 mM malate in the presence of 10 mM glutamate, 8 mM succinate in the presence of 5 mM

glutamate and 1 mM NADH were used as oxidation substrates. During succinate and NADH oxidation, 3 mkM rotenone

was added to the incubation medium. During NADH oxidation, 0.06 mM CaCl2 was added to incubation medium. The

concentrations of inhibitors of the respiratory chain were antimycin A (A-A) (20 mkM), BHAM (1 mM), KCN (0.4

mM), and CSP 310 antiserum (1 mg/mL). Polarograms were used to calculate the rates of phosphorylative respiration

(state 3), nonphosphorylative respiration (state 4), respiration control by Chance-Williams (RC), and the ADP:O ratio

(Estabrook 1967). The concentration of mitochondrial protein was analyzed according to Lowry et al. (1951). All

experiments were performed on 3-6 separate mitochondrial preparations. The data obtained were analyzed statistically

and arithmetic means and standard errors were determined.

Results and discussion. We studied the mitochondrial respiratory-chain function of winter wheat during short-term low

temperature stress and hardening using different oxidation substrates. When using succinate and NADH as oxidation

substrates, rotenone, which blocks electron transfer through complex I of the mitochondrial respiratory chain, was added

the mitochondrial-incubation medium. When using malate as oxidation substrate, winter wheat mitochondria isolated

from control seedling shoots were well coupled (Table 1). After short-term low-temperature stress, the rates of state-3

and state-4 respiration increased by 19.2 % and 43.8 %, respectively, and the respiratory-control coefficient (RC)

decreased (17.3 %) when compared to the control (Table 1). This data shows that these mitochondria were uncoupled.

On the other hand, mitochondria isolated from hardened winter wheat seedling shoots had a lower rate of state-3 and

state-4 respiration than the control mitochondria and changes in their RC coefficient and ADP:O ratio were to a lesser

degree (13 % for state 3 and 11 % for state 4) (Table 1). When succinate was used as an oxidation substrate, we found

that neither short-term low-temperature stress nor cold hardening influenced the degree of coupling of isolated mitochon-

109

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

dria (Table 1).

When NADH was

the oxidation

substrate, results

were similar to

those of succinate;

no significant

difference between

mitochondria

isolated from

control, stressed,

and hardened

shoots (Table 1).

Thus, cold stress

caused significant

changes only in the

activity of malate-

oxidizing mito-

chondria but did

not influence

succinate- and

NADH-oxidizing mitochondria. Short-term cold stress caused more pronounced changes in mitochondria energetic

activity then cold hardening.

The participation of the main cytochrome and alternative pathways in mitochondrial respiration was studied by

adding an oxidation substrate, mitochondria, and ADP to the polarographic cell. When mitochondria were in state-4

respiration, antimycin A, BHAM, and anti-CSP 310 antiserum or KCN were added to the polarographic cell. We found

that malate-oxidizing mitochondria isolated from control, stressed, and hardened seedling shoots differed in their

reaction to inhibitor addition. Antimycin A in addition to control mitochondria caused ~ 50 % decrease of oxygen

consumption. In mitochondria isolated from stressed plants, this treatment caused only ~ 30 % decrease (Fig. 1A). Cold

shock caused ~ 20 % increase of antimycin A-resistant mitochondrial respiration. In mitochondria isolated from hard-

ened plants, addition of antimycin A caused ~ 65 % decrease in oxygen consumption. Consequent addition of BHAM to

mitochondria isolated from control and hardened plants inhibited oxygen consumption up to 25 % from state-4 respira-

tion but in mitochondria isolated from stressed plants, this treatment inhibited oxygen consumption only up to 33 % (Fig.

1A). Therefore, we can conclude that in control mitochondria about 25 % of the respiration is antimycin A- and BHAM-

resistant and that this part of mitochondria respiration increased during short-term low-temperature stress but was at the

level of the control plants during cold hardening. The residual mitochondrial oxygen consumption was fully inhibited by

consequent addition of anti-CSP 310 antiserum or KCN, so we can conclude that this residual respiration is involved

with CSP 310 function.

Adding antimycin A to succinate-oxidizing mitochondria caused ~ 90 % inhibition of oxygen consumption (Fig.

1B). The consequent addition of BHAM to control mitochondria fully inhibited oxygen consumption. Despite the

absence of cold-shock influence on total mitochondrial activity (Table 1), this treatment caused an increase of antimycin

A-resistant respiration to ~ 20 % of that of state-4 respiration. Consequent addition of BHAM nearly inhibited mito-

chondrial respiration (Fig. 1B). Cold hardening caused an increase of antimycin A-resistant respiration to ~40 % that of

state-4 respiration. This antimycin A-resistant respiration also was nearly inhibited by BHAM addition (Fig. 1B). We

conclude that in succinate-oxidizing winter wheat mitochondria only two electron-transport pathways function, the main

cytochrome pathway and an alternative antimycin A-resistant oxidase. Both cold shock and especially cold hardening

caused an increase in this alternative pathway.

In NADH-oxidizing control winter wheat mitochondria, the addition of antimycin A caused ~ 80 % decrease of

oxygen consumption (Fig. 1C). Consequent BHAM addition fully inhibited oxygen consumption in control mitochon-

dria, but this addition and even the consequent addition of anti-CSP 310 antiserum did not fully inhibit oxygen consump-

tion in mitochondria isolated from stressed plants. The residual respiration in this case was about 10 %. Based on these

data, we concluded that in succinate- and NADH-oxidizing mitochondria the main part of respiration depends on the

functioning of the main cytochrome respiratory chain pathway (77 % and 91 %, accordingly) but only ~ 50 % of

respiration depends on this pathway function in malate-oxidizing mitochondria.

Table 1. The energetic activity of winter wheat mitochondria isolated from control (1), stressed

(2), and hardened (3) shoots analyzed using different oxidizing substrates. Data are presented as

mean + standard error, n = 6–32.

Rate of oxygen uptake,

nMol O2/min/mg of protein Respiration

Substrate Variant State 3 State 4 control ADP:O

10 mM Malate + 1 82.6 + 1.7 32.0 + 1.7 2.60 + 0.15 2.65 + 0.12

10 mM glutamate 2 98.6 + 5.6 46.1 + 2.9 2.15 + 0.06 2.23 + 0.05

3 55.5 + 5.6 24.7 + 2.9 2.25 + 0.06 2.33 + 0.05

8 mM Succinate + 1 66.9 + 1.7 45.5 + 1.8 1.48 + 0.15 1.80 + 0.12

5 mM glutamate 2 69.1 + 5.6 47.5 + 2.9 1.47 + 0.06 1.62 + 0.05

3 63.5 + 3.9 44.4 + 2.7 1.51 + 0.09 1.56 + 0.03

1 mM NADH 1 109.5 + 5.3 96.4 + 5.6 1.14 + 0.04 1.05 + 0.19

2 105.1 + 4.2 83.6 + 6.1 1.27 + 0.06 1.05 + 0.06

110

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Wheat mitochondria

have different electron trans-

port pathways. One is an

alternative KCN- and antimy-

cin A-resistant oxidase. In

addition to this pathway,

different types of uncoupling

proteins recently were found in

plant mitochondria. The plant

stress protein CSP 310 is one

(Voinikov et al. 1998). Data

obtained from inhibitor

analyses agree with that about

the influence of exogenous

CSP 310 on different mito-

chondrial respiratory-chain

complex function (Grabelnych

et al. 2001). The effect of CSP

310 addition to isolated plant

mitochondria was detected at

complex I function but was not

detected in the functioning of

other respiratory chain com-

plexes. Now, we can show that

the main contribution to

mitochondrial respiration of the

CSP 310-pathway that was

inhibited by anti-CSP 310

addition was detected during

malate oxidation (25 %).

Because antimycin A

addition blocks electron

transfer through Q-cycle, i.e.,

inhibits the main cytochrome

respiratory chain pathway, we

can conclude that residual

mitochondrial respiration

depends on the functioning of

alternative pathways. There-

fore, during malate oxidation,

the main cytochrome pathway

contributes ~ 50 % to the total

mitochondria respiration. The

residual 50 % depends on

alternative oxidase (25 %) and

CSP 310 (25 %) functioning

(Table 2). Cold shock caused

about a two-fold decrease in

the main cytochrome pathway

and increased the contribution

of alternative pathways. On

the other hand, cold hardening

caused an increase in the

cytochrome pathway contribu-

Fig. 1. The effect of different respiratory chain inhibitors on state-4 oxygen uptake

by winter wheat mitochondria isolated from control, stressed (short-term cold

stress), and hardened (hardening) shoots. The inhibitors were added in the

sequence shown reading left to right.

A. 10 mM malate in the presence of 10 mM glutamate used as a substrate of

oxidation (M+SE, n = 3–32).

B. 8 mM succinate in the presence of 5 mM glutamate used as a substrate of

oxidation (M+SE, n = 3–19).

C. 1 mM NADH used as a substrate of oxidation (M+SE, n = 5–14).

111

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

tion and decreased the contribution of alternative

pathways in mitochondrial respiration (Table 2).

During succinate oxidation, the main part

of mitochondrial respiration depends on the main

cytochrome pathway function (about 90 %). At

the same time, during succinate oxidation, short-

term low-temperature stress and especially cold

hardening caused a significant increase of

alternative oxidase function. In NADH-oxidizing

winter wheat mitochondria isolated from control

plants, the main part of mitochondrial respiration

also depends on cytochrome pathway function

(about 77 %). Both cold shock and hardening did

not significantly influence the contribution of

different pathways in NADH-oxidizing mitochon-

dria. Concurrently, we also detected an increase

of residual mitochondrial respiration after antimy-

cin A and anti-CSP 310 addition up to 10 % in

these conditions (Table 2). In our opinion, this

fact could depend on the function of external

rotenone-insensitive and antimycin A-insensitive

NADH-cytochrome c reductase (Soole et al.

1990).

Based on our data, we conclude that the

contribution of the different mitochondrial

electron-transport pathways significantly depends

on the oxidized substrate. Short-term cold stress and cold hardening differ in their influence on the different electron

transport pathways in winter wheat mitochondria.

Acknowledgments. The work was possible, in part, with the support of the Russian Foundation of Basic Research

(projects 00-04-48093 and 02-04-06096) and the Siberian Division of Russian Academy of Sciences Youth Grant

(project 78).

References.

Estabrook RW. 1967. Mitochondrial respiratory control and the polarographic measurement of ADP:O ratio. Meth

Enzymol 10:41-47.

Grabelnych OI, Pobezhimova TP, Kolesnichenko AV, and Voinikov VK. 2001. Complex I of winter wheat mitochondria

respiratory chain is the most sensitive to uncoupling action of plant stress-related uncoupling protein CSP 310. J

Therm Biol 26:47-53.

Ito K. 1999. Isolation of two distinct cold-inducible cDNAs encoding plant uncoupling proteins from the spadix of

skunk cabbage (Symplocarpus foetidus). Plant Sci 149:167-173.

Kolesnichenko AV, Zykova VV, Grabelnych OI, Sumina ON, Pobezhimova TP, and Voinikov VK. 2000. Screening of

mitochondrial proteins in winter rye, winter wheat, Elymus, and maize with immunochemical affinity to the stress

protein 310 kD and their intramitochondrial localization in winter wheat. J Therm Biol 25:245-249.

Laloi M, Klein M, Riesmeier JW, Muller-Rober B, Fleury Ch, Bouillaud F, and Ricquier D. 1997. A plant cold-induced

uncoupling protein. Nature 389:135-136.

Lowry OH, Rosebrough NJ, Farr AL, and Randall RJ. 1951. Protein measurement with folin phenol reagent. J Biol

Chem 193:265-275.

Maia IG, Benedetti CE, Leite A, Turcinelli SR, Vercesi AE, and Arruda A. 1998. AtPUMP: an Arabidopsis gene

encoding a plant uncoupling mitochondrial protein. FEBS Lett 429:403-406.

Moller IM and Rasmusson AG. 1998. The role of NADH in the mitochondrial matrix. Trends Plant Sci 3:21-27.

Murayama S and Handa H. 2000. Isolation and characterization of cDNAs encoding mitochondrial uncoupling proteins

in wheat: wheat UCP genes are not regulated by low temperature. Mol Gen Genet 264:112-118.

Nantes IL, Fagian MM, Catisti R, Arruda P, Maia IG, and Vercesi AE. 1999. Low temperature and aging-promoted

expression of PUMP in potato tuber mitochondria. FEBS Lett 457:103-106.

Table 2. The contribution of cytochrome pathway (Cyt) or

alternative pathways with cyanide-resistant alternative oxidase

(AltAOX), CSP 310 (AltCSP310), and outer NADH-dehydrogenase

(NADHouter) to total respiration of winter wheat mitochondria in

control conditions (1), during short-term cold stress (2), and

during hardening (3) using different oxidizing substrates. The

contribution is expressed as a percent of the respiratory rate in

state 4.

Percent contribution

Variant Cyt AltAOX AltCSP310 NADHouter

10 mM malate in the presence of 10 mM glutamate.

1 48.8 26.0 25.2 0.0

2 28.2 38.8 33.0 0.0

3 64.5 10.4 25.1 0.0

8 mM succinate in the presence of 5 mM glutamate.

1 91.6 6.6 1.8 0.0

2 78.3 13.1 8.6 0.0

3 61.6 33.4 4.9 0.0

1 mM NADH.

1 77.2 21.7 0.0 1.1

2 79.4 10.7 0.0 9.9

112

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Pobezhimova TP, Voinikov VK, and Varakina NN. 1996. Inactivation of complex I of the respiratory chain of maize

mitochondria incubated in vitro by elevated temperature. J Therm Biol 21:283-288.

Ricquier D and Bouillaud F. 2000. The uncoupling protein homologues: UCP1, UCP2, UCP3, StUCP and AtUCP.

Biochem J 345:161-179.

Sluse FE and Jarmuszkiewicz W. 2002. Uncoupling proteins outside the animal and plant kingdoms: functional and

evolutionary aspects. FEBS Lett 510:117-120.

Soole KL, Dry IB, and Wiskich JT. 1990. Oxidation of NADH by plant mitochondria: kinetics and effects of calcium

ions. Physiol Plantarum 78:205-210.

Soole KL and Menz RI. 1995. Functional molecular aspects of the NADH dehydrogenases of plant mitochondria. J

Bioenerg Biomembr 27:397-406.

Takumi S, Tomioka M, Eto K, Naydenov N, and Nakamura C. 2002. Characterization of two non-homoeologous

nuclear genes encoding mitochondrial alternative oxidase in common wheat. Genes Genet Syst 77:81-88.

Vanlerberghe GC and McIntosh L. 1997. Alternative oxidase: from gene to function. Ann Rev Plant Physiol Plant Mol

Biol 48:703-734.

Voinikov V, Pobezhimova T, Kolesnichenko A, Varakina N, and Borovskii G. 1998. Stress protein 310 kD affects the

energetic activity of plant mitochondria under hypothermia. J Therm Biol 23:1-4.

The use of linoleic acid as an oxidation substrate by winter wheat mitochondria.

O.I. Grabelnych, T.P. Pobezhimova, A.V. Kolesnichenko, and V.K. Voinikov.

Free fatty acids (FFA) are effective uncouplers of oxidative phosphorylation depending on their protonophoric activity,

which causes a significant increase in the conductance of the inner mitochondrial membrane. Some data shows that

saturated FFA has less influence on mitochondrial membrane potential then unsaturated FFA (Penzo et al. 2002). In

addition, saturated FFA can regulate mitochondrial uncoupling protein activity (Jezek et al. 1997; Jarmuszkiewicz et al.

1998; Costa et al. 1999; Hourton-Cabassa et al. 2002) and even expression of these proteins (Muzzin et al. 1999;

Sbrassia et al. 2002).

The major FFA catabolic pathway in the cell is β-oxidation, which results in acetyl-CoA that can be completely

oxidized by cell to CO2 and H2O via the Kreb’s Acid Cycle. Intermediates of this cycle are the main mitochondrial

respiration substrate (Schulz 1991). The FFA β-oxidation activity of this pathway significantly increases upon seed

germination but dramatically decreases after photosynthesis establishment and during vegetative growth (Masterson and

Wood 2000). FFA was used as an oxidation substrate during the very early stages of sunflower and lettuce seed germina-

tion (Salon et al. 1988; Raymond et al. 1992) and in potato storage organs (Theologis and Laties 1980). At the same

time, data on the capability of wheat-seedling mitochondria to use FFA as an oxidation substrate and about the participa-

tion of different mitochondrial electron transport pathways in this process are lacking.

Thus, the aim of this study the function of winter wheat mitochondria during oxidizing of FFA and the participa-

tion of different mitochondrial electron-transport pathways in this process.

Materials and methods. Three-day-old, etiolated shoots of winter wheat cultivar Zalarinka were germinated on moist

paper at 26°C. Mitochondria were extracted from seedlings shoots by differential centrifugation (Pobezhimova et al.

1996). The isolated mitochondria were resuspended in the following medium: 40 mM MOPS–KOH buffer (pH 7.4),

300 mM sucrose, 10 mM KCl, 5 mM EDTA, and 1 mM MgCl2. Mitochondrial activity was recorded polarographically

at 27°C using a closed-type, platinum electrode in a 1.4-ml cell (Estabrook 1967). The reaction mixture contained 125

mM KCl, 18 mM KH2PO4, 1 mM MgCl2, and 5 mM EDTA, pH 7.4. Malate (10 mM) in the presence of glutamate (10

mM) and linoleic acid (0.056–750 mkM) were used as oxidation substrates. The concentrations of inhibitors of respira-

tory chain were rotenone (3 mkM), antimycin A (A–A) (20 mkM), BHAM (1 mM), and CSP 310 antiserum (1 mg/ml).

Polarograms were used to calculate the rates of phosphorylative respiration (state 3), nonphosphorylative respiration

(state 4), respiration control by Chance-Williams (RC), and the ADP:O ratio (Estabrook 1967). The concentration of

mitochondrial protein was analyzed by Lowry method (Lowry et al. 1951). All the experiments were performed on three

separate mitochondrial preparations. The data obtained were analyzed statistically and arithmetic means and standard

errors determined.

113

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Results and discussion. The amount of total FFA in winter wheat mitochondria is about 15 ng/mg of mitochondrial

protein (0.056 mkM) and increases to ~40 ng/mg (0.15 mkM) after short-term cold shock (Vojnikov et al. 1983). In our

experiments, we used physiological concentrations of FFA and higher concentrations (1-750 mkM).

In the first set of experiments,

linoleic acid (LA) was added to malate

oxidizing mitochondria in state 4 (Fig.

2, 1). We found that LA did not

influence mitochondrial oxygen uptake

in the range of 0.056–5 mkM. At 10

mkM, LA increased oxygen uptake by

25 %. At 20 mkM, a 87 % increase of

oxygen uptake was detected. Further

increases in the LA concentration in

the mitochondria incubation medium

(20–60 mkM) did not cause further

increases in state-4 respiration. On the

other hand, adding 100 mkM or more

LA caused at least a three-fold increase

in mitochondrial oxygen uptake with a

maximum at 500 mkM. The addition

of 100 mkM LA caused an increase in

the level of state-4 respiration up to

that of state-3 respiration.

Similar results were obtained

when the oxidizing of LA was the only

oxidation substrate for mitochondria (Fig. 2, 2). Physiological FFA concentrations and concentrations up to 5 mkM did

not cause an increase in oxygen uptake by winter wheat mitochondria. At the same time, at a concentration of 10 mkM,

mitochondrial oxygen uptake up to 43 % was detected. Higher LA concentrations caused increases in oxygen uptake by

mitochondria. The maximum oxygen uptake by winter wheat mitochondria was at LA concentration of 500 mkM. The

rate of uncoupled respiration (Fig. 2, 1) and the rate of linoleic acid-supported respiration (Fig. 2, 2) were equal; 50 mkM

LA.

Our data show that wheat mitochondria can successfully use linoleic acid as respiration substrate. Therefore,

we were interested in determining what mitochondrial electron-transport pathways participate in this process. By

looking at the influence of different electron-transport pathway inhibitors on oxygen uptake during 100 mkM LA

oxidation, we found that different mitochondrial electron-transport pathways participate in this process. The data

indicate that ~31 % of oxygen consumption was inhibited by the addition of antimycin A, ~34 % was inhibited by

BHAM addition, ~33 % was inhibited by rotenone addition, and 30 % was inhibited by anti-CSP 310 addition.

During the oxidizing of LA, our data show that electrons can transfer oxygen through all branches of the

electron-transport chain. Because rotenone is a complex-I inhibitor, the part of mitochondrial respiration that is not

inhibited by its addition could deal with the functioning of complex II and different rotenone-insensitive, internal NADH

dehydrogenases (Moller 1997).

Antimycin A addition blocks electron transport through complex III and, after this treatment, only alternative

CN-resistant oxidase (Vanlerberghe and McIntosh 1997) and CSP 310 (Kolesnichenko et al. 2002) still function. These

results agree with data on the influence of BHAM, which is an inhibitor of alternative CN-resistant oxidase and anti-CSP

310 antiserum, and its addition inhibits oxygen consumption dependent on CSP 310 function. Therefore, the LA-

dependent increase in oxygen consumption is involved with the functioning of all branches of mitochondrial electron

transport chain, both phosphorylative and nonphosphorylative.

Hermesh et al. (1998) used very high concentrations (0.5–2 mM) of FFA when studing mitochondrial energetic

activity and proposed that FFA effects depend on the FFA-dependent uncoupling of oxidative phosphorylation. We have

shown that LA concentrations higher than 50 mkM mitochondria change their metabolism to oxidizing LA as an oxida-

100

120

140

160

00,06 0,15 1 5 10 20 30 40 50 60 75 100 150 200 500 750

Linoleic acid, mkM

Rate of oxygen uptake,

nMol O 2 / min/ mg of protein

Fig. 2. The influence of linoleic acid on the rate of oxygen uptake in

state-4 (1) and linoleic acid-supported oxygen uptake (2) of winter wheat

mitochondria. 1. 10 mM malate in the presence of 10 mM glutamate used

as a substrate of oxidation (M + SE, n = 3) and 2. linoleic acid used as a

substrate of oxidation (M + SE, n = 3).

114

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

tion substrate, because the rate of LA-supported respiration becomes equal to the uncoupled rate after the addition of LA

respiration during malate oxidation. The function of the main cytochrome pathway in such conditions could depend on

oxidative phosphorylation uncoupling because FFA uncoupling activity causes an increase of oxygen consumption. In

addition to this pathway, other alternative electron-transport pathways function during LA oxidation. Based on our data,

winter wheat mitochondria can use LA as an oxidation substrate. Linoleic acid oxidation in these conditions depends on

the functioning of all electron-transport pathways that exist in plant mitochondria.

Acknowledgment. This work was performed, in part, with the support of the Siberian Division of Russian Academy of

Sciences Youth Grant (project 78).

References.

Costa ADT, Nantes IL, Jezek P, Leite A, Arruda P, and Vercesi AE. 1999. Plant uncoupling mitochondrial protein

activity in mitochondria isolated from tomatoes at different stages of ripening. J Bioenerg Biomembr 31:527-533.

Estabrook RW. 1967. Mitochondrial respiratory control and the polarographic measurement of ADP:O ratio. Methods

Enzymol 10:41-47.

Hermesh O, Kalderon B, and Bar-Tana J. 1998. Mitochondrial uncoupling by long chain fatty acyl analogue. J Biol

Chem 1273:3937-3942.

Hourton-Cabassa C, Mesneau A, Miroux B, Roussaux J, Ricquier D, Zachowski A, and Moreau F. 2002. Alteration of

plant mitochondrial proton conductance by free fatty acids. Uncoupling protein involvement. J Biol Chem

277:41533-41538.

Jarmuszkiewicz W, Almeida AM, Sluse-Goffart CM, Sluse FE, and Vercesi A. 1998. Linoleic acid-induced activity of

plant uncoupling mitochondrial protein in purified tomato fruit mitochondria during resting, phosphorylating, and

progressively uncoupled respiration. J Biol Chem 273:34882-34886.

Jezek P, Costa ADT, and Vercesi AE. 1997. Reconstituted plant uncoupling mitochondrial protein allows for proton

translocation via fatty acid cycling mechanism. J Biol Chem 272:24272-24278.

Kolesnichenko AV, Pobezhimova TP, Grabelnych OI, and Voinikov VK. 2002. Stress-induced protein CSP 310: a third

uncoupling system in plants. Planta 215:279-286.

Lowry OH, Rosebrough NJ, Farr AL, and Randall RJ. 1951. Protein measurement with Folin phenol reagent. J Biol

Chem 193:265-275.

Masterson C and Wood C. 2000. Mitochondrial b-oxidation of fatty acids in higher plants. Physiol Plantarum 109:217-

224.

Moller IM. 1997. The oxidation of cytosolic NAD(P)H by external NAD(P)H dehydrogenases in the respiratory chain

of plant mitochondria. Physiol Plantarum 100:85-90.

Muzzin P, Boss O, and Giacobino JP. 1999. Uncoupling protein 3: its possible biological role and mode of regulation in

rodents and humans. J Bioenerg Biomembr 31:467-473.

Penzo D, Tagliapietra C, Colonna R, Petronilli V, and Bernardi P. 2002. Effects of fatty acids on mitochondria: implica-

tions for cell death. Biochim Biophys Acta 1555:160-165.

Pobezhimova TP, Voinikov VK, and Varakina NN. 1996. Inactivation of complex I of the respiratory chain of maize

mitochondria incubated in vitro by elevated temperature. J Therm Biol 21:283-288.

Raymond P, Spiteri A, Dieuaide M, Gerhardt and Pradet A. 1992. Peroxisomal b-oxidation of fatty acids and citrate

formation by a particulate fraction from early germinating sunflower seeds. Plant Physiol Biochem 30:153-162.

Salon C, Raymond P, and Pradet A. 1988. Quantification of carbon fluxes through the tricarboxylic acid cycle in early

germinating lettuce embryos. J Biol Chem 263:12278-12287.

Sbrassia P, D’Adamo M, Leonetti F, Buongiorno A, Silecchia G, Basso MS, Tamburrano G, Lauro D, Federici M,

Daniele ND, and Lauro R. 2002. Relationship between plasma free fatty acids and uncoupling protein-3 gene

expression in skeletal muscle of obese subjects: in vitro evidence of a causal link. Clinical Endocrinol 57:199-207.

Schulz H. 1991. Beta oxidation of fatty acids. Biochim Biophys Acta 1081:109-120.

Theologis A and Laties GG. 1980. Membrane lipid breakdown in relation to the wound-induced and cyanide-resistant

respiration in tissue slices. Plant Physiol 66:890-896.

Vanlerberghe GC and McIntosh L. 1997. Alternative oxidase: from gene to function. Ann Rev Plant Physiol Plant Mol

Biol 48:703-734.

Vojnikov VK, Luzova GB, and Korzun AM. 1983. The composition of free fatty acids and mitochondrial activity in

seedlings of winter cereals under cold shock. Planta 158:194-198.

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Questioning the possible role of D-amino acids in wheat seedlings.

N.I. Rekoslavskaya, R.K. Salyaev, V.M. Sumzova, T.V. Kopytina, and A.M. Sobenin.

The D isomers of different amino acids (alanine, tryptophan, aspartate, glutamate, proline, and other amino acids) and

their derivatives have been detected in plants (Bell 1980), but their possible physiological functions are unknown in

plants. The presence of nonproteinogenic, D-amino acids in seeds and seedlings is believed to protect plant tissues from

pathogens and parasites (Bell 1980).

D-amino acids are actively synthesized by bacteria and low fungi (Davies 1977). Alanine racemase is of great

importance to bacteria because it supplies them with D-alanine from available L-alanine. Therefore, alanine racemase

may be a key enzyme in the synthesis of the protective peptidoglucan layer of the cell wall. In some cases, the D-amino

acids are abundant (Vicario et al. 1987).

Another mechanism by which D-amino acids are formed involves D-amino acid aminotransferase, which

produces a diversity of D-amino acids. Perhaps the synergistic action of the two enzymes racemase and D-amino acid

transferase accounts for the large amount of different D-amino acids that appear in bacterial cells and plant seedlings.

D-alanine and its dipeptide, D-alanyl-D-alanine, make up a considerable part of the nitrogen pool and probably

play a significant part in regulation of nitrogen metabolism in bacteria. D-amino acids are not toxic in plants, perhaps

because of neutralization via malonylation, acetylation, and glycosylation followed by compartmentalization in the

vacuole. The bonding of D-amino acids with malonyl or acetyl moyeties may be hydrolyzed and reveal amino acids in

intact form.

D-alanine and its derivatives in pea seedlings appeared during germination and disappeared on the 8th day of

growth (Ogawa et al. 1973). D-alanyl-D-alanine and D-alanylglycine were found in rice seedlings and leaves, respec-

tively (Manabe 1986; Manabe and Ohira 1983). Free and bound D-aspartic and D-glutamic acids were determined in

pea seedlings (Ogawa et al. 1977). The N-malonyl-D-tryptophan content increased in leaves of tomato, potato, wheat,

and other species during wilting and after drought during the period of recovery after osmotic stress (Rekoslavskaya et

al. 1988).

All of these data would seem to indicate that synthesis of D-amino acids and their further conversion have

ontogenetic, physiologic, and ecologic significance that is still unknown. As for N-malonyl-D-tryptophan, an acceptable

hypothesis is that it functions as a precursor of the plant hormone indoleacetic acid, IAA (Rekoslavskaya et al. 2002). In

reality, D-tryptophan has been demonstrated in a number of cases to be as active or even more active than L-tryptophan

as an auxin substitute (Rekoslavskaya 1986).

Using D-tryptophan as an IAA precursor illustrates the idea that pools of amino acids for nonprotein synthesis

can be created by means of the conversion of L-amino acids to D-amino acids. Direct competition for the amino acid

between nonprotein syntheses and protein synthesis occurs in the process of growth and development.

Thus, the appearance of D-amino acids in plants apparently is nonrandom, uncontrolled, and physiological

meaningless event, but the physiological significance of D-amino acids remains largely unclear and needs detailed study.

We have investigated the content of amino acids in wheat seedlings in relation with some enzyme activities of amino

acids metabolism different from protein biosynthesis have been done. The specific activity of racemase, transaminase,

and UDPG-transferase were estimated in wheat seedlings during the study.

Materials and methods. The spring wheat cultivar Scala was used in this study. Procedures to determine racemase and

transaminase activities were as described by Rekoslavskaya et al. (2002). UDPG-transferase activity was determined

according to the modified method primarily described by Kowalczyk and Bandurski (1991). Briefly, 21 g of leaf, 44 g of

stem, 5.6 g of young kernel, and 35.1 g of root tissue of green wheat shoots were harvested, ground with mortar and

pestle in liquid nitrogen, and extracted with the buffer containing 0.25 M HEPES, 5 mM EDTA Na2, 0.1 %

mercaptoethanol, and 0.025 % Triton X-100, pH 8.5. One mg of phenylmethylsulfonylfluoride was added to the ground

material at the time of extraction in order to prevent protease activity. The homogenate was passed through four layers

of cheesecloth and centrifuged at 10,000 x g for 20 min at 4oC. The activity of UDPG-transferase was estimated in the

116

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Table 3. Substrate specificity of the enzymes of amino acid metabolism, % to

conversion of tryptophan (Try). Experiments were repeated at least twice.

L- or D- D-amino acid L-amino acid Etioplast Cytosol

amino acid oxidase oxidase racemase racemase Transaminase

Try 100 100 100 100 100

Ala 135 128 583 112 239

Pro 36 43 350 106 152

Met 104 — 310 106 —

Phe 116 129 101 103 124

Val 83 — 455 102 160

Asp 78 — 197 101 116

Asn 65 128 30 100 0

Thr 50 — 480 98 —

Ser 74 71 449 96 454

Arg 43 52 141 96 —

Cys 45 73 179 95 —

Leu 117 130 331 95 146

His 41 84 66 94 96

Glu 48 58 102 94 125

Tyr 95 85 99 90 —

iLeu 86 91 317 89 —

supernatant fraction of each sample. The reaction mixture contained as the substrate 5 mmol of indoleacetic acid (IAA),

5 mmol of UDPG as the cofactor, and in order to prevent the ribosomes activity, 10-4 M CaCl2 were added to 1 ml of

supernatant. The reaction mixture was then incubated for 16 hours at 37oC. The reaction was stopped by adding of 1 ml

of isopropanol. The activity of UDPG-transferase was determined as nmoles of substrate converted during 1 h/mg of

protein. The IAA glucose ester content was determined after passing of reaction mixtures through a DEAE-cellulose

(acetate form) minicolumn (10 x 20 mm) in 6 ml of eluates of 50 % isopropanol. The Ehrlich reagent was used in order

to determine IAA-glucose content with calibration curve made with IAA. A D-amino acids kit was used (Sigma, USA).

L-Amino acids were from Reachim (Russian Federation). The content of amino acids were determined on an amino-acid

analyzer AAA-1 (Microtechna, Czech Republic).

Results and discussion. The amino-acid content of 7-day-old seedling are presented in Fig. 3. The amino acids Glu,

Ala, Val, Pro, Leu, and iLeu had the highest content of > 200 mg/g of fresh weight. The content of Asp was next highest,

but the other amino acids were present at levels below 100 nmol/g of fresh weight. Free Try did not contribute any

significant content of free amino acids, but the sum of free and bound malonyl D-Try content was nearest to the content

of Glu or even greater in seedlings sustaining wilting; 890 nmol/g of fresh weight (Rekoslavskaya et al. 1988).

The appearance of D-amino acids, and especially D-Try, during germination and growth of etiolated seedlings

in the dark was shown previously (Rekoslavskaya et al. 2002). The activity of tryptophan racemase was found in the

cytosol and etioplast fractions of

wheat seedlings. The enzyme

was isolated and some biochemi-

cal characteristics were studied,

but the substrate specificity was

broader and racemase used other

amino acids as substrates (Table

3).

As shown in Table 3,

the chiralic purity of D- or L-

amino acids used were estimated

with D-amino acid oxidase from

hog kidneys (Sigma, USA) or

with L-amino acids oxidase from

snake venom (Sigma, USA).

When D- or L-amino acids were

treated with the enzyme prepara-

tion from wheat seedlings

prepared as described earlier

(Rekoslavskaya et al. 2002), we

observed higher enzyme activi-

ties than in the case of either D-

or L-tryptophan. For example,

the specificity to Ala, Thr, Val, or

Ser was about 5.8 or 4.5 times

higher than to Try. The activity of transaminase was higher if Ala, Ser, Val, and some other amino acids were exploited

in the study in comparison to Try. Therefore, it might be concluded that there was racemase and transaminase with broad

substrate activities in wheat seedlings with some preference to amino acids structurally related to Ala.

About half of the amino acids is in the form of D-enantiomers in etiolated wheat seedlings. The content of D-

and L-amino acids in 7-day-old wheat seedlings were 233.4 ± 34.0 and 194.8 ± 9.2 nmol/100 seedlings, respectively. We

found two pools of amino acids in growing wheat seedlings and question why half of the amino acids in wheat are in a

nonproteinogenic form that is not available for the synthesis of protein.

We tried to explain the appearance of D-Try in wheat seedlings as a creation of nonproteinogenic storage form

for the precursor for IAA biosynthesis when the growth was fast during germination. Nevertheless, free Try was

essential but not the predominant amino acid in wheat seedlings (Fig. 3). Thus, the role of other D-amino acids still

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

remained obscure. We searched for other

explanations for the possibility of using

nonproteinogenic amino acids for wheat

seedlings, which they possess in order to

survive in ecologically unfavorable

conditions.

Amino acids might be used in

the formation of plant lectins or

phytoagglutenines. Plant lectines may

play the role of antibodies against soil

bacteria and fungi and participate in the

defense response of young seedlings

because the localization of lectins was

found in embryos and other parts of plant.

The binding action of amino acids to a

sugar moiety was provided by UDPG-

transferase. UDPG-transferases are a

widespread and abundant enzyme family

with broad substrate specificity. As a

model system, we used IAA as a substrate in order to evaluate the activity of

UDPG-transferase in wheat shoots, because IAA is a derivative of the amino

acid Try and closely related to it in indole and side chain structure (Table 4).

The activity of UDPG-transferase was high in all parts of the wheat

plant. Therefore, wheat seedlings have a highly active system for balancing the

IAA level that was produced by rapid synthesis from D-Try. As a whole, the

IAA biosynthesis and its metabolism is sufficiently intense to provide for the fast

growth of etiolated seedlings during the heterotrophic period in order to emerge

from the soil and initiate photosynthesis. The D-amino acids, which are not

involved in protein biosynthesis, might participate in the protection of young

seedlings from pathogens, bacteria, and fungi by this very unique manner of joining with glucose or another sugar

moiety. This objective will be the subject of following experiments.

References.

Bell EA. 1980. Non-protein amino acids in plants. In: Enc Plant Physiol 8:403-432.

Davies JS. 1977. Occurrence and biosynthesis of D-amino acids. In: Chemistry and biochemistry of D-amino acids

4:1-27.

Kowalczyk S and Bandurski RS. 1991. Enzymic synthesis of 1-O-(indol-3-ylacetyl)-b-D-glucose. Purification of the

enzyme from Zea mays, and preparation of antibodies to the enzyme. Biochem J 279:509-514.

Manabe H. 1986. Effect of exogenous D-alanine on D-alanyl-D-alanine content in leaf blades of Oryza australiensis

Domin. Plant Cell Physiol 27:573-576.

Manabe H and Ohira K. 1983. Effect of light irradiation on the D-alanylglycine content in rice leaf blades. Plant Cell

Physiol 24:1137-1142.

Ogawa T, Fukuda M, and Kei S. 1973. Occurrence of N-malonyl-D-tryptohan in pea seedlings. BBA 297:60-69.

Ogawa T, Kimoto M, and Sasaoka K. 1977. Identification of D-aspartic acid and D-glutamic acid in pea seedlings.

Agric Biol Chem 41:1811-1812.

Rekoslavskaya NI. 1986. Possible role of N-malonyl-D-tryptophan as an auxin precursor. Biol Plantarum 28:62-67.

Rekoslavskaya NI, Markova TA, and Gamburg KZ. 1988. Appearance of N-malonyl-D-tryptophan in excised leaves

during wilting. J Plant Physiol 132:86-89.

Rekoslavskaya NI, Yurieva OV, Shainyan BA, Kopytina TV, and Salyaev RK. 2002. Wheat racemase and the role of

stereoisomers of N-malonyltryptophan during seed germination. Ann Wheat Newslet 48:141-143.

Vicario PP, Green BG, and Katzen HM. 1987. A single assay for simultaneously testing effectors of alanine racemase

and/or D-alanine. J Antibiot 40:209-216.

Fig. 3. The content of free amino acids in wheat seedlings and en-

dosperm at 7 days.

Table 4. The specific activity of

UDPG-transferase in wheat

shoots, nmol of IAA glucosyl

ester/mg of protein/h.

Leaves 9.08 ± 0.04

Stems 12.18 ± 0.22

Young kernels 7.92 ± 0.53

Roots 15.43 ± 0.18

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Changes in the aquaporin content in winter wheat membranes after deadaptation.

G.B. Borovskii, A.Yu. Yakovlev, S.V. Vladimirova, and V.K. Voinikov.

In the past decade, we have discovered that water transport in cells is not directly through membranes but through

numerous channels in the membranes. These channels are formed by proteins adhering to aquaporins. Aquaporins are

found in the plasma and vacuolar membranes in animal and plant cells (Maurel 1997; Connolly et al. 1998). By regulat-

ing the degree of aquaporin phosphorylation, the cell controls the permeability of a membrane to water (Maurel et al.

1997; Kjellbom et al. 1999) and changes in the amount of these proteins shift the range of regulation. During adaptation

to low temperature, membrane permeability increases and water migrates into the intercellular spaces during freezing

(Alberdi and Corcuera 1991). This increase in permeability very likely is associated with an increase of aquaporins in

the membranes. We expect the reverse during deadaptation in the spring. To date, changes in the quantity of water-

channel proteins during deadaptation of overwintered plants has not been investigated.

Materials and methods. The crowns and leaves of winter wheat plants of the cultivar Irkutskaia ozimaia were used in

this study. This genotype is winter hardy and

highly productive under the severe climatic

conditions of eastern Siberia (Borovskii et al.

2001). Crowns, leaves, and soil monoliths

with plants were sampled in the field in

January. Crowns and leaves were used for

membrane-fraction isolation. The remaining

plants in the monoliths were left at room

temperature for 1 month under natural

illumination for de-adaptation. After 1 month,

the crowns and leaves were harvested and the

membrane fraction isolated. We identified

aquaporins inside the microsomal membrane

fraction, because antibodies demonstrated a

high degree of specificity (Fig. 4).

Wheat membranes were isolated by centrifugation at 105,000 g for 1 h. Proteins were dissolved in a sample-

loading buffer at 65°C. Proteins were separated by SDS–PAGE using a mini-Protean II PAGE cell (Bio-Rad, U.S.A.)

according to the manufacturer’s instructions.

Western blotting and immunodetection were as

described by Timmons and Dunbar (1990) using

anti-PIP (plasmalemma-intrinsic protein) and anti-

TIP (tonoplast-intrinsic protein) primary antibodies

(1:1000 dilution), kindly provided by Dr. A.

Schaeffner (Institute of Biochemical Plant Pathol-

ogy, München, Germany) and Dr. C. Maurel

(Institut des Sciences Végétales, Gif-sur-Yvette,

France), respectively.

Results and discussion. We observed a decrease

in aquaporins in both leaves and crowns after

deadaptation of winter wheat (Fig. 5). Plasmale-

mma and tonoplast aquaporins decreased. This

data supports the hypothesis that decreases in

membrane water permeability occur after spring

deadaptation. We assume that the permeability of

membrane to water decreases in plants, because

permeability is associated closely with freezing

resistance. Alternatively, changes in the aquaporin

content of the membrane could be connected with

the start of the next stage plant development after

overwintering plants reinitiate growth.

Fig. 4. Specificity using antibodies against two types of

aquaporins, TIP (tonoplast-intrinsic protein, A) and PIP

(plasmalemma-intrinsic protein, B). Immunoblotting of the proteins

obtained from tonoplast (1) and plasmalemma (2). Arrows and

numerals between the images indicate molecular weight markers.

Fig. 5. Changes in aquaporin content in the membrane

proteins of winter wheat plants after a 1-month deadaptation.

The microsomal fraction was separated from leaves (1, 2) and

crowns (3, 4). Plants were taken from field (1, 3) or left for

30 days under ambient conditions (2, 4). Proteins were

fractionated on 12 % SDS–PAGE gels. Aquaporins were

identified by specific antibodies against TIP (tonoplast-

intrinsic protein, A) and PIP (plasmalemma-intrinsic protein,

B). Arrows and numerals between the images indicate the

molecular size markers.

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

The aquaporin content culminates after development in the autumn; water exits the cell during freezing. We

know that some aquaporins are strongly induced by ABA (Kaldenhoff and Eckert 1999). This fact indirectly confirmed

our results, because ABA content is high during winter adaptation and decreases under deadaptation in the spring.

Activation of the water channels is useful to expel water and entrance inside under extreme thawing. In our opinion,

regulating the action of water channels under the freezing in the external spaces of the cell is the same mechanisms that

takes place under the water stress (Kjellbom et al. 1999), by stress-increasing of Ca2+ content in the cytoplasm. After

winter, a high aquaporin content is dangerous because Ca2+ content in the cytoplasm increases under any stress.

Changes in the permeability of cell membranes to water are very important for plant adaptation to freezing. The

importance requires a tight control of permeability. Our results suggest that aquaporins are involved in adaptation of

wheat to winter and deadaptaion in spring.

Acknowledgments. The work has been supported by the Russian Foundation of Basic Research (projects 02-04-48728

and 02-04-48599). We sincerely thank Dr. A. Schaeffner and Dr. C. Maurel for gift of antibodies.

References.

Alberdi M and Corcuera LJ. 1991. Cold acclimation in plants. Phytochem 30:3177-3184.

Borovskii GB, Stupnikova IV, Peshkova AA, Dorofeev NV, and Voinikov VK. 2001. Ann Wheat Newslet 47:179-185.

Connolly DL, Shanahan CM, and Weissberg PL. 1998. The aquaporins. A family of water channel proteins. Internat J

Biochem Cell Biol 30:169-172.

Kaldenhoff R and Eckert M. 1999. Features and function of plant aquaporins. J Photochem Photobiol B Biol 52:1-6.

Kjellbom P, Larsson C, Johansson I, Karlsson M, and Johansson U. 1999. Trends Plant Sci 4:308-314.

Maurel C, Kado RT, Guern J, and Chrispeels MJ. 1995. Phosphorylation regulates the water channel activity of the

seed-specific aquaporins a-TIP. EMBO J 14:3028-3035.

Maurel C. 1997. Aquaporins and water permeability of plant membranes. Ann Rev Plant Physiol Plant Mol Biol

48:399-429.

Timmons TM and Dunbar BS. 1990. Protein blotting and immunodetection. Methods Enzymol 182:679-688.

Using urea nitrogen for the nutrition of spring wheat under adverse temperatures.

A.K. Glyanko, N.V. Mironova, and G.G. Vasilieva.

Introduction. Urea is used widely in agriculture and is highly competitive with, and under certain conditions superior

to, mineral forms of N fertilizers in its effect on yield and quality. For example, urea contributes to a greater accumula-

tion of protein, gluten, and indispensable amino acids in wheat grain and other cereals during grain formation and

maturation (Finney et al. 1957; Pavlov 1967; Schlehuber and Tacker 1967; Slukhai and Zrazhevsky 1971; Mitrofanov et

al. 1973; Fox et al. 1986). Urea is taken up rapidly and metabolized by plants (Mokronosov et al. 1966; Pavlov 1967;

Andrews et al. 1984). Urea increases the permeability of membranes and tissues and enhances the uptake, transferal, and

reutilization of nutrients in plants (Mitrofanov et al. 1973; Turley and Ching 1986).

The mechanisms by which ammonium fertilizer and urea nitrogen affect plant metabolism are different

(Tishenko et al. 1991). Thus, the role of urea as a N fertilizer has been studied in relatively sufficient detail, but the

influence of adverse environmental factors on plant nutrition and physiology by this form of nitrogen have not. Over the

last decade, researchers have had a great interest in studying the physiological response of plants to the nitrate and

ammonium forms of N under stress conditions of salinity, low temperature, drought, and inadequate illumination

(Chandra et al. 1986; Hubick 1990; Leidi et al. 1991; Gruz et al. 1993; Glyanko 1995).

Our results are derived from studying physiology of nutrition with urea nitrogen when spring wheat plants were

exposed to a late spring frost (–6, –7°C) and low soil temperature (> 0°C) to compared to using the mineral forms of

nitrogen.

Material and methods. Plant material and growth conditions. Soft spring wheat plants of the cultivar Skala were

grown in containers (eight plants/container) in a growth chamber at the Siberian phytotron (Irkutsk, Russia) at a tempera-

ture of 19 ± 1°C/15 ± 1°C (day/night), illuminated by DRL-700 incandescent lamps. The light intensity was 14 ± 0.5

kLx with a 16-hour daylength. Infrared radiation from the lamps was suppressed by a water screen. The plants were

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

grown using a sand–soil mixture with a small amount of total nitrogen (0.009 %). Macro- and microlements were

supplied at half the normal rate into enameled containers filled with dry soil (Grodzinsky and Grodzinsky 1973).

Watering was by weight with distilled water up to 70 % of the moisture capacity of the soil. To guard against any

nitrification of the ammonium, the nitrification inhibitor 2-chlor-6-trichlormetyl pyridine (N-serve) was introduced into

the containers at 1 % of the N dose.

Conditions of the artificial frost. A spring frost condition between –6 and –7°C was produced in a refrigerating

chamber of the phytotron once the plants had reached the three-leaf stage. The chamber was not illuminated during the

frost period. Temperature in the chamber was controlled automatically under a preset program (Kurets 1974). The

program provided for a gradual decrease in temperature within the chamber from the optimum temperature (19 ± 1°C) to

0°C (at the rate of 1°/12 min), followed by a decrease to the minimum subzero temperature (–6 and –7°C) at the rate of

1°/22 min. After a 1.5-hour exposure to temperatures between –6 and –7°C, the temperature was raised to 0°C at the rate

of 1°/12 min. The temperature was raised from 0°C to the optimum temperature at the same rate. The total time of

exposure of the plants to subzero temperature was 6 hr, of which 1.5 hour corresponds to the minimum subzero tempera-

ture. The relative air humidity within the chamber was 85–90 % during the frost. The containers with plants were

placed in holes in plastic foam to avoid freezing the soil during the frost. One and one-half hours after the end of the

frost (the temperature in the chamber was raised to 19°C), both control and experimental plants were fed through their

roots with a mixture of three forms of N, one of which contained labeled 15N. The extra nutrition schemes were variant I,

15NH414NO3 + 14N – urea; variant II, 14NH415NO3 + 14N – urea; and variant III, 14NH414NO3 + 15N – urea.

In variant I, where the label was in the NH4 group, 25.9 mg 15N were introduced in each container and the

enrichment of 15NH4NO3 was 95.31 weight percent of 15N; in variant II, 24.3 mg 15N with an enrichment of NH415NO3 of

89.66 weight percent of 15N; in variant III, 52.3 mg 15N were introduced with a urea enrichment of 93.84 weight percent

of 15N. The total amount of nitrogen that was introduced into the vessels during the extra nutrition was 106.4 mg in the

first two variants and 101.3 mg in variant III.

Soil temperature reduction. To reduce the temperature in the root zone, containers with plants were placed in

thermal chambers through which water at 5 ± 1°C and 19 ± 1°C was passed, maintaining the required soil temperature

(Kurets 1974).

Chemical analyses. Protein in the triturated leaves was precipitated with trichloroacetic acid. Nucleic acids

and other soluble compounds were removed from the protein precipitate (Klyachko et al. 1971). The protein was

digested in concentrated sulfuric acid with a catalyst, selenium (Se). Protein nitrogen was distilled by the micro-Kjeldale

method and determined by the titrimetric method of Ermakov et al. (1987). Samples were analyzed for enrichment of

15N by means of a mass-spectrometer MI-1309. The content of labeled N in samples was determined by a formula for

isotopic dilution (Korenkov 1977). The atomic percent of 15N was converted to weight percent of 15N (Korenkov 1977).

The activities of glucose-6-phosphate dehydrogenase (G-6-PD) and malate dehydrogenase (MD) were determined using

biochemical tests (Boehringer and Soehne GmbH Mannheim, Germany) in cell-free, unpurified root extracts. Urease

activity was determined according to Bollard et al. (1968), and the protein in cell-free preparations was quantified

according to Lowry et al. (1951). The biological and analytical repeatability of assays was fivefold and threefold,

respectively. Results are represented as the arithmetic mean with a standard error. The confidence level of the differ-

ences was evaluated by the Student t-test (tst). Least significant difference for comparing treatment means at the 0.95

probability level.

Results and discussion. Effect of late spring frost. Of 195 plants that underwent frosts, 64 (32 %) had one damaged

leaf, eight had two damaged leaves, and three plants died. Thus, 38 % of the plants showed visually observable dam-

ages.

The sample for quantifying protein was made from the laminas of two plants having no visible damage. The

plants did not show any substantial differences in protein accumulation in their leaves during the first 9 days after the

frost, the absolute content in both control and experimental plants increased by a factor of 1.6 to 1.7. Labeled N is

incorporated into leaf protein at a different rate depending on the form of N-fertilizer (Table 5). For example, 9 days

after the frost, 552.9 µg 15N from urea, 137.0 µg from the ammonium group, and 73.8 µg from the nitrate group were

determined in the protein of the control plants. The percentage of labeled N utilization by the plants from fertilizers

amounted to 1.06, 0.53, and 0.30, for urea, ammonium, and nitrate, respectively. During frost, this remains regular

(Table 5). The difference is that a short exposure to subzero temperature promotes the incorporation into protein of the

121

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

label from urea. When compared to control plants, the label incorporation is 115 and 150 % at 3 and 9 days after the

frost, respectively (differences at td>tst).

Label incorporation into protein 1, 3, and 9 days

after the frost also is stimulated from the 15NO3 group. The

confidence level of the differences between the control and

the assays are very high (P > 0.99). With regard to the effect

of frost on the incorporation of the label from the 15NH4

group, a reliable decrease in 15N incorporation into protein on

day 3 is observed (P > 0.95), whereas the differences are

unreliable at 1 and 9 days after the frost (td<tst).

The utilization of labeled N from different forms of

N on day 9 after the frost was 1.58, 0.57 and 0.48%, from

urea, ammonium, and nitrate, respectively. Thus spring

wheat seedlings predominantly utilize urea N in synthesizing

the protein. Temperature stress has a stimulating effect on

this process. The control and experimental plants did not

differ in absolute protein N content in the leaves (Table 5),

suggesting that, during increased catabolic processes such as

after frost, plants are able to shift the state of decay-synthesis

of proteins toward the latter through an intense utilization of

urea N.

The predominant utilization of urea from the mixture

of three forms of N can probably be explained by a couple of

factors. First, the relatively easy uptake of urea by roots.

Second, the fast transport of urea (or its products) to aerial

organs and subsequent use in metabolism.

In comparison with mineral forms of N (NO3- and

NH4+), the mechanism of urea uptake by plants is not yet

understood (Van Beusichem and Neeteson 1982). We

anticipate that urea, as a neutral compound, is taken up by

root cells with a minimum expenditure of energy and a high

proportion is transported to aerial organs in an unchanged

form. Urease activity in wheat roots and seedling leaves

when the plant roots receive extra nutrition of urea provides

some evidence. Activity of urease in leaves increases by a

factor of 2.9, whereas enzyme activity in the roots is uncer-

tain.

Chen and Ching (1988) induced leaf urease activity

when barley plants are sprayed with urea solution. They

detected urease isozymes, which were synthesized only

during the period of an abrupt increase in enzyme activity.

Our data indicate that spring wheat seedlings contain a

sufficiently active constitutive form of urease in their roots

and a less active form in leaves (medium without N). Under

the influence of extra nutrition of plant roots with urea,

urease activity changes little in roots but increases abruptly in

leaves. The latter is likely to be associated with the de novo

synthesis of enzyme.

The reasons for stimulating the uptake of label from urea as an effect of frost are unclear. We determined the

urease and nitrate reductase activity in wheat leaves as an effect of the frost (within 1 and 3 days) and found that the

activity of both enzymes was enhanced. However, we only can explain the presence in cells of a sufficient number of

Table 5. The effect of a late spring frost on 15N uptake by leaf protein during nutrition of wheat from a mixture of different forms of nitrogen.

N-protein content is from the dry weight of two plants.

Within 1 day Within 3 days Within 9 days

N protein 15N content N protein 15N content N protein 15N content

Assay and content Weight % (N protein, content Weight % (N protein, content Weight % (N protein,

variant (mg) (excess 15N) g) (mg) (excess 15N) g) (mg) (excess 15N) g)

Control 4.19 ± 0.36 0.33 14.5 ± 0.70 3.65 ± 0.20 0.95 36.5 ± 2.41 6.08±0.46 2.14 137.0 ± 5.02

I; frost 3.77 ± 0.05 0.42 16.6 ± 0.73 4.11 ± 0.22 0.67 29.0 ± 1.53 6.17 ±0.15 2.29 148.8 ± 7.41

Control 4.46 ± 0.07 0.02 0.90 ± 0.06 4.23 ± 0.33 0.24 11.3 ± 0.83 7.41 ±0.12 0.89 73.8 ± 8.01

II; frost 4.01 ± 0.22 0.05 2.20 ± 0.17 4.19 ± 0.18 0.32 15.0 ± 1.03 6.88 ± 0.18 1.50 115.6 ± 8.15

Control 4.19 ± 0.09 1.12 50.2 ± 4.31 4.23 ± 0.11 4.32 195.5 ± 8.13 7.62 ± 0.16 6.78 552.9 ± 27.00

II; frost 4.45 ± 0.41 1.09 51.9 ± 3.81 4.22 ± 0.15 4.99 225.3 ± 9.41 7.46 ± 0.04 10.37 827.9 ± 41.30

122

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

NH4+ ions needed for the synthesis of amino acids. The mechanism of the effect of low temperature on the transcription-

translation apparatus in leaves when plants are fed with different forms of nitrogen remains to be elucidated.

Reduced temperature effect of soil. Urea as fertilizer behaves in a peculiar fashion at low above-freezing temperatures

in the root zone. We found that after exposure to low temperature (5 ± 1°C), G-6-PD and MD activity increases in roots

by a significantly greater amount when the plants were fed with urea as compared to NO3– and NH4+. The activity of G-

6-PD in the roots by urea is stimulated by 7-fold, as opposed to 3.2- and 3.9-fold for the NO3- and NH4+ N-sources,

respectively. Under normal temperatures, enzyme activity in plants is higher with NO3 nutrition. The stimulating effect

of NO3 on enzymes of the pentose monophosphate pathway of carbohydrate oxidation has been reported (Givan 1979).

The activity of MD at near-freezing temperature increases in roots by 267, 167, and 136 % in variants with urea, NH4+,

and NO3–, respectively. At the optimum temperature in the root zone (19 ± 1°C), however, the activity of these enzymes

during urea nutrition of plants is lower when compared to variants with other nitrogen forms. A possible mechanism to

explaining the stimulation of the G-6-PD and MD activity under stress could be the dissociation of the multidimensional

forms of enzymes into simpler subunits having increased activity. The presence of electrophoretically different forms of

enzymes suggests that under different conditions in the medium the relationship of different molecular forms of enzymes

can change drastically (Petrova et al. 1985), which is responsible for the increase or decrease in enzyme activity.

We observed a greater stimulating of enzyme activity under low-temperature effect in the presence of urea. In

protein chemistry, urea is known as a dissociating agent of proteins (Zolkiewski et al. 1995). At low temperatures,

conditions that allow the penetration of urea to places where compartmentalizing of enzymes may be created in cells and

the molar concentration suffices to have a dissociating effect on enzymes. An alternative explanation for the activation

of the G-6-PD and MD enzymes could be an enhancement, at low temperature, of other processes such as anaplerotic

pathways for the assimilation of carbonic acid during the enzymatic decomposition of urea in plant cells. This pathway

involves enhancing the carboxylation processes with the participation of root phosphoenolpyruvate carboxylase and

other CO2-fixing enzymes resulting in products that are used in the Krebs cycle.

When urea is used to nourish plants in the root zone at low temperature, root growth is enhanced. According to

our data from a water-culture assay, the presence of urea as the only growth source in the nutrient solution causes

enhanced growth of plants if the temperature in the root zone was 5 ± 1°C. This effect of urea on root growth was not

observed in the root zone at the optimum temperature. This assay was repeated in soil-cultured plants. In this case,

nitrogen in the form of different fertilizers was introduced at 42 mg/kg soil (210 mg/container). All other elements were

introduced at one-half the normal concentration. Once seedlings appeared, containers with seedlings were placed in

different temperature conditions and the plants were grown until the third leaf appeared. At optimum soil temperature,

the plants reached the 3-leaf stage within 13–14 days; at low temperature this occurred with in 21–23 days.

Our results showed that at low soil temperature and optimum air temperature (19 ± 1°C), the root dry weight in

the variant with urea was higher when compared to plants grown with the other forms of N. The mean length of roots in

the variant with urea at both the low and optimum temperatures was greater when compared with the other N-sources

(Table 6). The root wet weight during urea nutrition under low temperature conditions in both water and soil culture

approaches or

exceeds that in

the variant

without N.

Nitrogen

deficiency and

phosphorus in

the medium is

known to

promote growth

of the plant root

system (Barber

1979), and the

presence of

these elements

leads to a

decrease in

Table 6. Wet weight of roots and of the aerial portion, and mean length of 15 spring wheat seed

lings as a function of soil temperature and N-form. Air temperature was the same for all variants,

19 ± 1°C.

Temperature

5 ± 1°C 19 ± 1°C

weight of weight of length of weight of weight of length of

roots aerial portion roots roots aerial portion roots

Variant (g) (g) (cm) (g) (g) (cm)

No N (control) 7.3 ± 0.07 2.8 ± 0.17 27 ± 0.6 6.4 ± 0.15 3.2 ± 0.31 29 ±0.5

Ca(NO3)25.6 ± 0.33 4.4 ± 0.39 24 ± 0.4 4.4 ± 0.33 4.7 ± 0.33 19 ± 0.5

(NH4)2SO46.8 ± 0.12 4.2 ± 0.30 24 ± 0.3 4.1 ± 0.07 5.2 ± 0.10 18 ± 0.7

(NH2)2CO 8.7 ± 0.08 6.0 ± 0.52 26 ± 0.5 4.6 ± 0.24 5.5 ± 0.41 22 ± 0.1

123

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

intensity of growth. In this case, during urea nutrition under low soil temperature conditions, plant roots behave as in the

variant without N.

The mechanism responsible for enhancing root growth in the absence of N (or phosphorus) in the medium is

unknown. Barber (1979) suggests that a stem-connected feedback mechanism causes an increase in root growth. Such a

mechanism could be a hormonal imbalance in wheat roots during nutrition of plants with urea and other forms of N.

According to our data, the relation between indoleacetic acid and abscisic acid in root tissues of wheat seedlings varies

according to the form of N and soil temperature (Glyanko 1995). Lips (1997) also reported that variation in the balance

between abscisic acid and cytokinins in roots during nitrate and ammonium nutrition has an effect on the growth of roots

and aerial organs and contributes to adaptation of plants to stress effects (salinization or moisture deficiency). Thus,

enhancement of root growth in conditions of near-freezing temperatures is effected under the influence of urea, and

activation of urea N in protein molecules as an effect of frost is manifested by the adaptive and reparative changes in

wheat plants induced by the form of N.

References.

Andrews RK, Blakeley RL, and Zerner B. 1984. Urea and urease. In: Advances in Inorganic Biochemistry (Eichhora

GL and Marzilli LG eds), Elsevier, New York. 6:245-283.

Barber SA. 1979. Uptake of nutrients from soil by plant roots. Physiol Biochem Cultiv Plants 11:209-217 (in Russian).

Bollard EJ, Cook AK, and Turner NA. 1968. Urea as sole source of nitrogen for plant growth. 1. The development of

urease activity in Spirodella eligorriza. Planta 83:1-12.

Chandra R, Radhuver P, and Sirohi GS. 1986. Influence of moisture stress and nitrogen on growth and yield of pea and

sorghum. Ann Arid Zone 25: 225-231.

Chen Y and Ching TM. 1988. Induction of barley leaf urease. Plant Physiol 86:941-945.

Cruz C, Lips SH, and Martins-Loucao MA. 1993. Effect of root temperature on carob growth: nitrate versus ammo-

nium nutrition. J Plant Nutrit 16:1517-1530.

Ermakov AI, Arasimovich VV, and Yarosh NP. 1987. Methods of biochemical investigation of plants. VO

Agropromizdat, Leningrad Pp. 237-238 (in Russian).

Finney KF, Mayer JM, Smith FM, and Fryer HC. 1957. Effect of Pawnee wheat with urea solution on yield, protein

content and protein quality. Agron J 49:341-347.

Fox RH, Kern JM, and Piekieler WP. 1986. Nitrogen fertilizer source, and method and time of application effects on

no-till corn yields and nitrogen uptakes. Agron J 78:741-746.

Givan CV. 1979. Metabolic detoxification of ammonia in tissues of higher plants. Phytochem 18:373-383.

Glyanko AK. 1995. Nitrogen nutrition of wheat at low temperatures. Nauka, Novosibirsk (in Russian).

Grodzinsky AM and Grodzinsky DM. 1973. Concise manual on plant physiology. Naukova Dumka, Kiev (in Russian).

Hubick KT. 1990. Effects of nitrogen source and water limitation on growth, transpiration efficiency and carbon-isotope

discrimination in peanut cultivars. Aust J Plant Physiol 17:1413-1430.

Kurets VK. 1974. The Irkutsk phytotron. Nauka, Novosibirsk (in Russian).

Klyachko N, Yakovleva LA, and Kulaeva ON. 1971. Change in protein synthesis in the cotyledons of pumpkin in

connection with their age. Fiziologiya Rasteniy (Sov Plant Physiol) 18:1225-1231.

Korenkov DA. 1977. Methods of application of nitrogen isotope 15N in agricultural chemistry. Kolos Publishing House,

Moscow (in Russian).

Leidi EO, Soares MIM, Silberbush M, and Lips SH. 1991. Salinity and nitrogen nutrition studies on peanut and cotton

plants. J Plant Nutrit 15:591-604.

Lips SH. 1997. The role of inorganic nitrogen ions in plant adaptation processes. Rus J Plant Physiol 44:421-431.

Lowry OH, Rosenbrough NJ, Farr AL, and Randall RJ. 1951. Protein measurement with the Folin phenol reagent. J

Biol Chem 193:265-275.

Mitrofanov BA, Okanenko AS, and Pochinok KhN. 1973. Effect of urea spray application of winter wheat on intensity

of photosynthesis and grain quality. Physiol Biochem Cultiv Plants 5:232-238 (in Russian).

Mokronosov AT, Ilinych ZG, and Shukolyukova NI. 1966. Assimilation of urea by potato plants. Fiziologiya Rasteniy

(Sov Plant Physiol) 13:798-806.

Pavlov A. 1967. Protein accumulation in wheat and maize grains. Nauka, Moscow (in Russian).

Petrova OV, Kolosha OI, Mishustina PS, and Sukhareva IB. 1985. Enzyme form multiplicity and its modification in

winter wheat in the period of adaptation to low temperatures. Physiol Biochem Cultiv Plants 17:361-366 (in Rus-

sian).

Schlehuber AM and Taker BB. 1967. The growing of wheat. In: Wheat and wheat improvement (Reitz LP and

Quisenberry KS eds). American Society of Agronomy, Madison, WI. Pp. 140-198.

124

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Slukhai SI and Zrazhevsky MN. 1971. Increase of winter wheat grain quality under condition of irrigation. Physiol

Biochem Cult Plants 3:303-313 (in Russian).

Tishenko NN, Nikitin DB, Magomedov IM, and Moran E. 1991. Effect of nitrate and ammonium forms of nitrogen

fertilizers on sugarcane photosynthesis and growth parameters. Physiol Biochem Cultiv Plants 23:446-452 (in

Russian).

Turley RH and Ching TM. Physiological responses of barley leaves to foliar applied urea- ammonium nitrate. Crop

Sci 26:987-993.

Van Beusichem ML and Neeteson JJ. 1982. Urea nutrition of young maize and sugarbeet plants with emphasis on ionic

balance and vascular transport of nitrognous compounds. Neth J Agric Sci 30:317-330.

Zolkiewski M, Nosworthy NJ, and Ginsburg A. 1995. Urea-induced dissociation and unfolding of dodecametric

glutamine synthetase from Escherichia coli – calorimetric and spectral tudies. Protein Sci 4:1544-1552.

VAVILOV INSTITUTE OF GENERAL GENETICS, RUSSIAN ACADEMY OF

SCIENCES

Gubkin str. 3, 119991 Moscow, Russian Federation.

SHEMYAKIN AND OVCHINNIKOV INSTITUTE OF BIOORGANIC CHEMISTRY,

RUSSIAN ACADEMY OF SCIENCES2

Ul. Miklukho-Maklaya 16/10, Moscow, Russian Federation.

Isolation and characterization of antimicrobial peptides from Triticum kiharae.

T.I. Odintsova and V.A. Pukhalskiy (Vavilov Institute of General Genetics) and Ts.A. Egorov and A.K. Musolyamov

(Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry).

All living organisms have evolved mechanisms with which to defend themselves against pathogen attack. This innate

immunity involving the production of antimicrobial peptides is one of the most ancient and widespread defense strate-

gies. After defense peptides are produced by transcription and translation of a single gene, they can be delivered rapidly

after infection with a limited input of energy and biomass and display differential activity against different types of

microorganisms (Thomma et al. 2002). Different families of antimicrobial peptides have been identified, including

thionins, defensins, lipid-transfer proteins (LTPs), hevein-type peptides, and knottin-type peptides.

We hoped to identify the antimicrobial peptides in T. kiharae, which is highly resistant to most pathogens

infecting cultivated wheat. T. kiharae has been used in our laboratory in crosses to generate lines resistant to such fungal

pathogens as powdery mildew and brown rust.

Materials and methods. The peptide fraction was extracted from T. kiharae flour with 10 % acetic acid (flour to

solution ratio of 1:10) for 1 h at room temperature. The supernatant was lyophilized and subjected to chromatography.

The acid-soluble fraction was separated by gel-exclusion chromatography on a Sephacryl S-100 HR column using 10 %

acetonitrile containing 0.1 % TCA as eluent. The chromatographic fractions were tested for the antifungal activity

against several fungi (Helminthosporium sativum, Alternaria consortiale, Rhizoctonia solani, Botritis cinerea, and

Fusarium culmorum). The active fraction, which caused inhibition of fungal growth and morphological changes, was

separated by reversed-phase high-performance liquid chromatography (RP–HPLC). The HPLC-fractions were tested

against fungi and characterized by mass spectrometry (MS) and sequencing.

Results and discussion. Separation of acid-soluble peptides on a Sephacryl column produced six fractions designated

from A to G. Only fraction D exhibited antifungal activity against most fungi assayed. This fraction was further

separated by RP–HPLC. Several fractions were obtained. Their molecular masses were measured by MS, and N-

terminal sequences identified by automatic sequencing. The peptide masses separated by RP–HPLC are in Table 1.

The N-terminal sequences of all fractions were determined. Two fractions were identified: Fr. 4:

AXQASQLAVXASAILGGTKPSGE and Fr. 5: KSXXK/RSTL

125

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

The N-terminal sequence of

fraction 4 coincides with that of LTP;

however, three substitutions at positions 3,

4, and 5 have been observed (Garcia-

Olmedo et al. 1998). Plant LTPs are 90–

95 amino acid polypeptides that have been

identified (at a protein and/or cDNA

levels) in various tissues from a high

number of mono- and dicotyledonous

species. They were found to be distributed

throughout the plant. Antimicrobial

activity of LTPs has been reported for all

members of the family tested. The relative

activities of different LTPs vary between

pathogens, suggesting that they have some

degree of specificity. The mass of LTP

from T. kiharae is lower than that de-

scribed in the literature for other members

of this family.

According to the N-terminal sequence, fraction 5 corresponds to α/β purothionins. The toxicity of thionins to

plant pathogens is known from investigations into the susceptibility to wheat endosperm thionins of phytopathogenic

bacteria in the genera Pseudomonas, Xanthomonas, Agrobacterium, Erwinia, and Corynebacterium. Purified genetic

variants of these thionins differed in activity and showed some degree of specificity. Recent experiments in planta also

are indicative of a defense role for the thionins.

Other fractions obtained by RP–HPLC of T. kiharae peptides were heterogeneous; therefore, their sequencing

produced inconclusive results. Some low-molecular peptides were sequenced directly after the separation of the total

acetic-acid extract on an RP–HPLC column. The sequences obtained were TRQLSLRG and TRQLSPRG. Homologous

proteins were not found in the data banks, so their functions remain unknown.

These results indicate that T. kiharae possesses different types of antimicrobial peptides.

References.

Thomma B, Cammue B, and Thevissen K. 2002. Plant defensins. Planta 216:193-202.

Garcia-Olmedo B, Molina A, Alamillo J, and Rodriguez-Palenzuela P. 1998. Plant defense peptides. Biopolymers

(Peptide Science) 47:479-491.

Distribution of hybrid necrosis genes in common wheat cultivars of Australia.

V.A. Pukhalskiy, S.P. Martynov, and E.N. Bilinskaya.

We studied the necrosis genes in modern cultivars of spring common wheat of Australia. The distribution of hybrid

necrosis genes in the old local cultivars was first investigated by Tsunewaki and Hori (1967, 1968), who showed that the

Ne1 ne2 and ne1 ne2 genotypes prevailed by the end of the 19th and early in the 20th centuries. The available data on the

wheat cultivars of Australia and Oceania indicate that 25.4 % are of the Ne1 ne2 genotype, 18.9 % are ne1 Ne2, and 55.7

% are ne1 ne2 (Pukhalskiy et al. 2000). This genotype distribution resulted from nearly a century of breeding in Austra-

lia. We thought it interesting to investigate this parameter at the end of the 20th century.

Materials and methods. The necrotic genotype was analyzed in 48 Australian cultivars of spring common wheat. The

spring common wheat cultivars Marquillo (Ne1sNe1s ne2ne2 genotype) and Balaganka (ne1ne1 Ne2sNe2s) were used as

testers. Crosses were conducted under field conditions by standard procedures including emasculation and isolation of

spikes. The F1 and F2 hybrids were grown in the field. Hybrid necrosis traits were evaluated at different growth stages.

Table 1. Molecular mass of the RP-HPLC fractions obtained from the

fungicidal fraction D. Prevailing masses are indicated in bold.

Fraction number

1 23457

1,371.6 1,236.5 1,070.6 1,153.5 1,206.8 3,487.9

1,425.6 1,344.8 1,644.8 1,405.8 5,900.9

2,734.0 1,535.8 2,189.0 1,574.8 16,372.1

3,451.3 1,829.1 3,484.1 3,021.2

7,007.2 2,678.6 6,972.4 3,622.0

3,373.3 4,803.5

3,565.6 4,919.6

6,983.2

7,641.9

126

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Results and discussion. The distribution of the

different necrosis genotypes in Australian wheat

cultivars shows that breeding led to complete

elimination of the Ne1ne2 genotype (Table 2). If

we estimate the ratios of necrotic genotypes in all

46 cultivars (except for cultivars Beulah and Bt-

Schomburgk where the presence of the Ne2 gene is

problematic), the results are as follows: 76.1 % of

cultivars possess the ne1 ne2 genotype and the ne1

Ne2 genotype is found in 23.9 % of cultivars.

The ratios for wheats at the beginning of

the 20th century were different (Tsunewaki et al.

1967). Among 72 cultivars examined, the ne1 ne2

genotype was found in 57 (79.2 %) of the cultivars,

Ne1 ne2 in 14 cultivars (19.4 %), and ne1 Ne2 (1.4

%) only in one cultivar. The ne1 Ne2 genotype

was found in the cultivar Atlas (Tsunewaki et al.

1968). The authors did not indicate whether Atlas

is a winter or a spring cultivar. In all probability,

Atlas was one of the two winter wheat cultivars

studied.

We suppose that the observed changes in

the distribution of hybrid necrosis genes were due

to the Green Revolution and to the wide use of

CIMMYT material in the Australian breeding

programs.

Pedigree analysis of the Australian wheats

using the GRIS 3.5 (Martynov and Dobrotvorskaya

1993) shows the Brazilian landrace Turco as the

source of the Ne2 gene. In addition, this gene

could be derived from the Argentinian landrace

Barleta or the Japanese cultivar Norin 10, the donor

of the short-stem trait, which has the Ne2 gene

from the landrace Mediterranean through the old,

American cultivars Lancaster and Fultz.

Acknowledgment. The authors are grateful to

Michael MacKay, the curator of the Australian

collection of winter cereals, for the seeds of

modern Australian wheat cultivars used in this

study.

References.

Martynov SP and Dobrotvotvorskaya TV. 1993.

Breeding-oriented database on genetical

resources of wheat. Ann Wheat Newslet

39:214-221.

Pukhalskiy VA, Martynov SP, and Dobrotvorskaya

TV. 2000. Analysis of geographical and

breeding-related distribution of hybrid necrosis

genes in bread wheat (Triticum aestivum L.).

Euphytica 114:233-240.

Table 2. Genotype of necrosis genes identified in modern

Australian cultivars of common spring wheat.

AWCC number Cultivar Year of release Genotype

AUS-25046 Cunningham 1990 ne1 ne2

AUS-25139 Lillimur 1990 ne1 ne2

AUS-25418 Angas 1991 ne1 ne2

AUS-25292 Excalibur 1991 ne1 ne2

AUS-25648 Cadoux 1992 ne1 ne2

AUS-25468 Katunga 1992 ne1 ne2

AUS-27166 Pulsar 1992 ne1 Ne2

AUS-25598 Amery 1993 ne1 ne2

AUS-25567 Beulah 1993 ne1?

AUS-25929 Darter 1993 ne1 ne2

AUS-25568 Goroke 1993 ne1 ne2

AUS-25868 Houtman 1993 ne1 Ne2

AUS-25571 Ouyen 1993 ne1 ne2

AUS-25927 Rowan 1993 ne1 ne2

AUS-25923 Stiletto 1993 ne1 ne2

AUS-25597 Stretton 1993 ne1 ne2

AUS-25869 Sunmist 1993 ne1 Ne2

AUS-25870 Sunstate 1993 ne1 Ne2

AUS-25928 Swift 1993 ne1 ne2

AUS-25557 Tasman 1993 ne1 ne2

AUS-25924 Trident 1993 ne1 ne2

AUS-25925 Vectis 1993 ne1 ne2

AUS-25619 Wellstead 1993 ne1 ne2

AUS-25600 Bt-Schomburgk 1994 ne1?

AUS-25575 Cascades 1994 ne1 ne2

AUS-26161 Datatine 1994 ne1 ne2

AUS-25931 Sunland 1994 ne1 ne2

AUS-26160 Tammin 1994 ne1 ne2

AUS-24350 Yarralinka 1994 ne1 ne2

AUS-25558 Pelsart 1994 ne1 ne2

AUS-26169 Tern 1994 ne1 ne2

AUS-26192 Leichhardt 1995 ne1 Ne2

AUS-25607 Arnhem 1996 ne1 Ne2

AUS-27194 Carnamah 1996 ne1 ne2

AUS-27193 Cunderdin 1996 ne1 Ne2

AUS-27189 Kalannie 1996 ne1 ne2

AUS-27188 Perenjori 1996 ne1 ne2

AUS-27191 Petrel 1996 ne1 ne2

AUS-27192 Sunlin 1996 ne1 ne2

AUS-27199 Yanac 1996 ne1 ne2

AUS-27190 Tailorbird 1996 ne1 Ne2

AUS-25601 Frame 1997 ne1 ne2

AUS-25602 Barunga 1997 ne1 ne2

AUS-27203 Krichauff 1998 ne1 ne2

AUS-27647 Diamondbird 1997 ne1 Ne2

AUS-27694 Baxter 1998 ne1 Ne2

AUS-27660 Goldmark 1998 ne1 Ne2

AUS-27661 Silverstar 1998 ne1 ne2

127

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Tsunewaki K and Hori T. 1967. Distribution of necrosis genes in wheat. IV. Common wheat from Australia, Tibet and

Northern Europe. Jap J Genet 42:245-250.

Tsunewaki K and Hori T. 1968. Necrosis genes in common wheat varieties from Australia, Tibet and Northern Europe.

Wheat Inf Serv 26:22-27.

N.I. VAVILOV RESEARCH INSTITUTE OF PLANT INDUSTRY

42, B. Morskaya Str., St. Petersburg, 190000, Russian Federation.

Genealogical analysis of Russian and Ukrainian winter wheat resistant to common bunt.

S.P. Martynov and T.V. Dobrotvorskaya.

Common bunt is one of most serious diseases of bread wheat. This disease is distributed in many regions of the Russian

Federation including the Northern Caucasus, Central Black Soil region, Volga region, and Non-Black Soil zone. Resis-

tance to common bunt in winter wheat was measured by comparing groups of resistant and susceptible cultivars from the

Russian Federation and Ukraine using a genealogical approach.

Data on winter bread wheat cultivars were taken from the database GRIS 3.5 of the Information and Analytical

System of Wheat Genetic Resources (Martynov and Dobrotvorskaya 2000). A set of 199 cultivars with known resis-

tance/susceptibility to common bunt and known pedigrees were divided into resistant (Table 1) and susceptible (Table 2)

groups.

Tracing expanded pedigrees with the aid of the GRIS program has established the probable donors and sources

of resistance to common bunt (Table 1). Except for eight cultivars for which it was impossible to identify the source of

resistance, the source of resistance to common bunt 36 cultivars (82 %) was from local sources mainly A. glaucum via

PPG-599, Crimean, Odessa local cultivar (LV-Odessa) via Zemka, Eliseevskaya rye, and Yaroslav emmer. Other culti-

vars (18 %) received resistance genes both from local and foreign sources; Florence (Bt3) and Oro (Bt4, Bt7). A number

of cultivars have ambiguous estimations of resistance to bunt (marked by an * in Table 1). For example,

Bezenchukskaya 380 is considered resistant in the Lower Volga region but susceptible in other areas. Moskovskaya 70

and 642, Moskovskaya nizkostebelnaya, Chaika, and Yantarnaya 50 are classified as resistant, but data from State

Varietal Trials indicates susceptibility. Skorospelka 1 and 3, from source data, and Odesskaya 12, from State Varietal

Trials, are resistant, but data from the Vavilov Institute identifies them as susceptible. We assume that the conflicting

data are a consequence of the different race compositions of local pathogen populations. Krivchenko (1984) has identi-

fied 37 different pathogen races. Analyzing the geographical distribution of the pathogen races, we identified two groups

appropriate to two conventional regions; north and south of latitude 49°N. Races 1, 9, 15, 17, and 20 comprised the

southern group and 2, 14, 16, 25, 31, 34, and 37 were specific to the northern group. Races 6 and 11 were common to

both groups. We assume that the sources of resistance differ in southern and northern regions. Therefore, we analyzed

groups of resistant and susceptible cultivars divided into southern and northern subgroups (see Tables 1 and 2). Among

the cultivars of the southern area, the basic sources of resistance are the Odessa local variety (LV-Odessa) via Zemka,

selection from Crimean (CI-1435), and foreign sources via Brevor and CIMMYT cultivars. In the northern subgroup,

the number of sources of resistance is more limited; A. glaucum via PPG 599 and Eliseevskaya rye.

In a three-way ANOVA of the matrixes of ancestor contribution (Table 3), we investigated the resistance (factor

A) with two classes (resistance and susceptibility), the region of origin (factor B) with two classes (south and north), and

the original ancestor or hypothetical source of resistance (factor C) with the number of classes (c = 11). The analyzed

sample included 52 resistant cultivars (including 23 from the southern and 29 from the northern regions) and 147

susceptible cultivars (including 88 from the southern and 59 from the northern regions). The data were transformed

through arcsines. The effects of all investigated factors and interactions, except for interaction (A x B) were highly

significant. Highly significant interactions (A x C), (B x C), and (A x B x C) indicate specific differences between the

distribution of the contributions of hypothetical sources of resistance in groups of resistant and susceptible cultivars

occurring from various regions. Differences in the race composition of regional populations of pathogen explain this

fact.

128

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Table 1. Donors of resistance to common bunt in Russian (RUS) and Ukrainian (UKR) winter wheat cultivars. Culti-

vars marked with an asterisk (*) have conflicting estimates of resistance by different authors.

Country Year of Hypothetical

of origin release Donor of resistance source of resistance

Cultivars bred south of 49°N latitude

Kooperatorka (Bt1) UKR 1929 Sel. Crimean Crimean

Stepnyachka-0496 UKR 1929 Sel. Banatka Kherson Unknown

Odesskaya-12* UKR 1947 Zemka LV-Odessa

Skorospelka-3-B RUS 1955 Kanred Crimean (CI 1435)

Deviz RUS 1978 Skorospelka-3-B, Narodnaya Crimean (CI 1435),

Narodnaya

Odesskaya polukarlikovaya UKR 1980 Odesskaya-12 LV-Odessa

Stepnyak UKR 1982 Odesskaya-12 LV-Odessa

Dneprovskaya-39 UKR 1984 Kanred Crimean (CI 1435)

Prikumskaya-79 RUS 1984 Penjamo-62 (Brevor, Florence, Oro, Yaroslav

Newthatch), Odesskaya-12 emmer, LV-Odessa

Dneprovskaya -1029 UKR 1989 Skorospelka -1* Crimean (CI-1435)

Dneprovskaya -710 UKR 1989 Odesskaya-12, LV- Odessa, Crimean (CI

Skorospelka -3B 1435)

Panna UKR 1990 Chaika*, Skorospelka -1* LV- Odessa, Crimean (CI

1435)

Donchanka UKR 1990 Odesskaya polukarlikovaya, LV- Odessa, Eliseevskaya

Odesskaya-12, RPG-434-154

Donshchina RUS 1992 Skorospelka -3*, Narodnaya Crimean (CI 1435),

Narodnaya

Odesskaya -133 UKR 1993 Brevor Florence, Oro

Donchanka 3 UKR 1995 Brevor Florence, Oro

Delta RUS 1999 Selkirk Crimean (CI 1435),

Yaroslav emmer

Knyajna RUS 1999 Selkirk (?) Crimean (CI 1435),

Yaroslav emmer

Prikumskaya 115 RUS 1999 Mida (U.S.), Odesskaya-12 Florence, LV-Odessa

Prima odesskaya UKR 2000 Odesskaya-12 (?) LV-Odessa

Zernogradka 10 RUS 2001 RPG-424-154, Eliseevskaya, Crimean

Skorospelka –3B (CI 1435)

Zarnitsa RUS 2002 Brevor Florence, Oro

Stanichnaya RUS 2002 Odesskaya-12, Brevor LV-Odessa, Florence, Oro

In the northern region, the contributions of A. glaucum and Eliseevskaya rye are higher in the group of resistant

cultivars. In the southern region, the Odessa local variety prevails among resistant cultivars (Table 4). In the northern

region, the contribution of LV-Odessa is higher in the group of susceptible cultivars, confirming the race specificity of

this resistance source. Yaroslav emmer, in the northern region, and foreign sources (Oro, Florence, Federation, and T.

timopheevii), in the south, are effective, although their contribution is not significant when compared with the group of

susceptible cultivars.

This analysis shows that number of sources of a vertical resistance to bunt used in the winter wheat-breeding

programs in the Russian Federation and Ukraine is not sufficient. The high number of genotypes with identical reaction

to bunt causes genetic uniformity in the cultivars. The narrowing of the genetic diversity from a few identical genes can

cause a change in the pathogen population and increase susceptibility on homogeneous genetic material.

Efficient horizontal (nonracespecific) resistance, which is shown as incomplete resistance to all races of a

pathogen and in varying degrees suppresses its development, also depends on the genetic diversity of the released

cultivars. A study of latent genetic diversity in winter wheat cultivars from the Russian Official List has shown that the

129

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Table 1 (continued). Donors of resistance to common bunt in Russian (RUS) and Ukrainian (UKR) winter wheat

cultivars. Cultivars marked with an asterisk (*) have conflicting estimates of resistance by different authors.

Country Year of Hypothetical

of origin release Donor of resistance source of resistance

Cultivars bred north of 49°N latitude.

Milturum-120 UKR 1929 Sel. Krasnokoloska Krasnokoloska (?)

Moskovskaya-3251 RUS 1929 Sel. Tavtukhi (T. durum) Unknown

RPG-27-36 RUS 1931 Eliseevskaya rye Eliseevskaya rye

RPG-434-154 RUS 1931 Eliseevskaya rye Eliseevskaya rye

PPG-599 (Btz)RUS 1948 Agropyron glaucum A. glaucum

Lgovskaya-873 RUS 1952 Unknown Unknown

Belotserkovskaya-198 UKR 1955 Unknown Unknown

PPG-99 (Btz)RUS 1964 PPG-599 A. glaucum

Kalininskaya-27 RUS 1965 A. glaucum A. glaucum

Kalininskaya-11 (Btz)RUS 1967 A. glaucum A. glaucum

Kharkovskaya-63 UKR 1969 Unknown Unknown

Zarya (Btz)RUS 1978 PPG-599 A. glaucum

Polukarlik-Mytnitskii UKR 1984 Schlanstedter (?) Unknown

Bezenchukskaya yubileinaya RUS 1984 RPG-434-154 Eliseevskaya rye

Yantarnaya-50* (Btz)RUS 1985 Zarya A. glaucum

Moskovskaya nizkostebelnaya* RUS 1990 Zarya A. glaucum

Inna RUS 1991 Zarya A. glaucum

Moskovskaya 642* RUS 1991 Zarya A. glaucum

Moskovskaya 70* RUS 1991 Zarya A. glaucum

Zvezda* RUS 1992 Ag. glaucum A. glaucum

Nemchinovskaya 25 RUS 1992 Zarya A. glaucum

Pamyati Fedina RUS 1993 Zarya A. glaucum

Bezenchukskaya 380* RUS 1994 RPG-434-154 Eliseevskaya rye

Smuglyanka RUS 1998 PV-18, Brevor, RPG-434-154 Florence, Oro, Yaroslav

emmer, Eliseevskaya rye

Povoljskaya 86 RUS 1999 Zarya A. glaucum

Moskovskaya 39 RUS 1999 Yantarnaya -50, Brevor A. glaucum, Florence,

Oro

Guberniya RUS 2000 Unknown Unknown

Tau RUS 2001 Selkirk Crimean (CI 1435),

Yaroslav emmer

Omskaya 4 RUS 2001 RPG-434-154 Eliseevskaya rye

overwhelming majority (96 %) of cultivars recommended for cultivation in the Russian Federation are the descendants of

Bezostaya 1 and/or Mironovsakaya 808. In the Central Black Soil zone and the Northern Caucasus and Middle and

Lower Volga regions, the genetic diversity is acceptable, whereas the Central Non-Black Soil and Volga-Vyatka regions

of the Russian Federation are characterized by low genetic diversity. The majority of cultivars recommended for these

regions are related at the full- and half-sib level.

A key problem of breeding for resistance to bunt is use of the new sources of resistance. In addition to the 11

known resistance genes (Bt1–Bt10 and BtZ), 11 new genes have now been identified. Ukrainian researchers have

identified six new genes; Bt11 from Sel. M-6623, Bt12 and Bt13 from Lutescens 6028, and Bt14 from Erythrospermum

5221 (Novokharka et al. 1990) and Bt15 and Bt16 from Ferrugineum 220/85 (Babayants and Dubinia 1990). CIMMYT

researchers have identified five new genes, which, unfortunately, have been given the same gene designations; Bt11

(from PI-554119), Bt12 (from PI-119333), Bt13 (from Thule III), Bt14 (from Doubbi), and Bt15 (from Carleton)

(Wilcoxson and Saari 1996). In addition, two presumably new genes in lines Erythrospermum 60-89 and Ferrugineum

124-88 were identified (Babayants et al. 1999). Some parental forms of Erythrospermum 5221, Ferrugineum 220/85,

130

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Erythrospermum 60-89, and Ferrugineum 124-88 are unknown, which does not enable pedigree analysis. We could

analyze the pedigree only of Lutescens 6028 and are now able to explain bunt resistance in this line.

Tracing the transmission of Bt-genes on the expanded pedigrees the has shown that Lutescens 6028 (Selection

101/Manella//Kavkaz) can have genes Bt1, Bt3, Bt4, Bt6, and Bt7 from Selection 101 (Fig. 1) that has the following

Table 2. Russian (RUS) and Ukrainian (UKR) winter wheat cultivars susceptible to common bunt.

Country of Year of Country of Year of

Cultivar origin release Cultivar origin release

Cultivars bred south of 49°N latitude.

Odesskaya 3 UKR 1938 Volgodar RUS 1990

Pervenets RUS 1938 Donetskaya 46 UKR 1990

Voroshilovskaya RUS 1939 Olimpiya 2 RUS 1990

Ferrugineum 622-2 RUS 1939 Khersonskaya 86 UKR 1991

Erythrospermum 161 RUS 1941 Odesskaya 117 UKR 1992

Novoukrainka 83 RUS 1945 Skifyanka RUS 1992

Krymskaya 1 UKR 1946 Tarasovskaya 87 RUS 1992

Gibrid 481 RUS 1948 Yubileinaya 75 UKR 1992

Osetinskaya 4 RUS 1950 Zernogradka 8 RUS 1993

Osetinskaya G-720 RUS 1950 Kolos Dona RUS 1993

Gibrid 491 RUS 1951 Odesskaya 120 UKR 1993

Kubanskaya 131 RUS 1951 Soratnitsa RUS 1993

Kubanskaya 24 RUS 1952 Sfera RUS 1993

Novoukrainka 84 RUS 1953 Fedorovka UKR 1993

Odesskaya 16 UKR 1953 Donskaya yubileinaya RUS 1994

Osetinskaya 3 RUS 1953 Eika RUS 1994

Yubileinaya Osetii RUS 1954 Otrada RUS 1994

Bezostaya 4 RUS 1955 Krasnodarskaya 90 RUS 1995

Lutescens 32 RUS 1965 Leda RUS 1995

Krasnodarskaya 33 RUS 1967 Nika Kubani RUS 1995

Skorospelka 35 RUS 1968 Azau RUS 1997

Donetskaya 5 UKR 1982 Aliza RUS 1997

Krasnodarskaya 57 RUS 1982 Zimorodok RUS 1997

Donskaya polukarlikovaya RUS 1983 Nak RUS 1997

Prikubanskaya RUS 1983 Odesskaya 267 UKR 1997

Dar Zaporojya UKR 1984 Viktoriya Odesskaya UKR 1998

Donetskaya 38 UKR 1984 Don 95 RUS 1998

Zaporojskaya 60 UKR 1984 Jirovka RUS 1998

Zirka UKR 1984 Zernogradka 9 RUS 1998

Olimpiya RUS 1984 Kroshka RUS 1998

Brigantina UKR 1986 Pobeda 50 RUS 1998

Prokofevka UKR 1986 Uskoryanka RUS 1998

Stepnaya 7 RUS 1986 Podarok Donu RUS 1999

Zamena RUS 1987 Dar Zernograda RUS 2000

Zimdar 4 RUS 1987 Donskoi mayak RUS 2000

Peresvet UKR 1987 Starnad 1 RUS 2000

Prometei UKR 1987 Tarasovskaya Ostistaya RUS 2000

Zimdar RUS 1988 Ermak RUS 2001

Birlik RUS 1989 Lira RUS 2001

Mriya Khersona UKR 1989 Prestij RUS 2001

Olviya UKR 1989 Rosinka Tarasovskaya RUS 2001

Prikumskaya 98 RUS 1989 Tarasovskaya 97 RUS 2001

Yunnat Odesskii UKR 1989 Deya RUS 2002

Albatros Odesskii UKR 1990 Selyanka RUS 2002

131

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Table 2 (continued). Russian (RUS) and Ukrainian (UKR) winter wheat cultivars susceptible to common bunt.

Country of Year of Country of Year of

Cultivar origin release Cultivar origin release

Cultivars bred north of 49°N latitude.

Hostianum 237 RUS 1929 Meshinskaya RUS 1989

Erythrospermum 20 430 UKR 1934 Mironovskaya 40 UKR 1989

Hostianum 122 76 RUS 1936 Mironovskaya 61 UKR 1989

Kievlyanka 156 UKR 1936 Omskaya ozimaya RUS 1989

Lesostepka 76 UKR 1937 Kharkovskaya 11 UKR 1989

Svea Pushkinskaya RUS 1937 Komsomolskaya 56 UKR 1990

Saratovskaya 46 131 RUS 1938 Nemchinovskaya 52 RUS 1990

Erythrospermum 118 RUS 1938 Polesskaya 87 UKR 1990

Lutescens 17 UKR 1940 Lgovskaya 167 RUS 1991

Lesostepka 75 UKR 1945 Nemchinovskaya 86 RUS 1991

Sekisovskaya RUS 1949 Kharkovskaya 90 UKR 1991

Lutescens 230 RUS 1951 Mironovskaya 27 UKR 1992

PPG 1 RUS 1951 Mironovskaya >AB8AB0O UKR 1992

PPG 186 RUS 1953 Bazalt RUS 1993

Veselopodolyanskaya 499 UKR 1954 Lutescens 9 RUS 1993

Mironovskaya 808 UKR 1963 Meshinskaya 2 RUS 1993

Mironovskaya N18;59=0O UKR 1971 Mironovskaya poluintensivnayaUKR 1993

Polesskaya 70 UKR 1974 Kharkovskaya 92 UKR 1993

Raduga RUS 1976 Chernozemka 212 RUS 1993

Akhtyrchanka UKR 1978 Bagrationovskaya RUS 1994

Moskovskaya 60 RUS 1979 Veselopodolyanskaya 203 UKR 1995

Mironovskaya 25 UKR 1980 Imeni Rapoporta RUS 1995

Nemchinovskaya 110 RUS 1980 Saratovskaya 90 RUS 1995

Drujba 2 UKR 1981 Kruiz RUS 1998

Lgovskaya 77 RUS 1981 Orenburgskaya 105 RUS 1998

Kinelskaya 4 RUS 1985 Orenburgskaya 14 RUS 1998

Mechta 1 UKR 1985 Kharkovskaya 96 UKR 1999

Polesskaya 85 UKR 1985 Malakhit RUS 2000

Shchedraya Polesya UKR 1987 Ershovskaya 11 RUS 2002

Volgogradskaya 84 RUS 1989

Table 3. Analysis of variance of the contribution of hypothetical sources of

resistance to common bunt for Russian and Ukrainian winter wheat cultivars.

Factor A is the group of resistant cultivars, factor B is the geographical

region of origin, and factor C is the ancestry. * = significance at P < 0.0001.

Source SS Df Ms F

General 71,643.4 2,155

Factor A 246.6 1 246.6 15.88*

Factor B 402.8 1 402.8 25.93*

Factor C 33,766.3 10 3,376.6 217.45*

Interaction (A x B) 5.4 1 5.4 0.35

Interaction (A x C) 597.8 10 59.8 3.85*

Interaction (B x C) 2,784.3 10 278.4 17.92*

Interaction (A x B x C) 1,029.6 10 102.9 6.63*

Error 32,810.6 2,112 15.5

cultivars and genes in its pedigree:

Rex (Bt1 and Bt7), Rio (Bt6), Oro (Bt4

and Bt7), Florence (Bt3), Burt (Bt1,

Bt4, and Bt6), and Brevor (Bt1, Bt3,

Bt4, and Bt6). Novokhatka et al.

(1990) could not explain the results of

segregation of resistance in crosses

between ‘Lutescens 6028/Bt4 (mono-

genic line)’ and ‘Lutescens 6028/(Bt6)

Rio’. The first cross segregated 74:26,

which corresponds to the theoretical

ratio 189:67 (r2 = 0.002) suggesting

four genes (one basic and three

duplicate-complementary genes

(Manjunath and Nadaf 1983). A

segregation of 57:58 was found in the

second cross, corresponding to a

theoretical 121:135 (r2 = 0.24) and

132

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

suggesting four genes

(two basic complemen-

tary and two duplicate-

complementary genes

(Manjunath and Nadaf

1983). Thus, we cannot

prove that the resistance

genes in Lutescens 6028

are nonallelic and

independent from

previously described

genes Bt1, Bt3, Bt4, Bt6,

and Bt7. The high level

of resistance in Lutescens

6028 may come from a

combination of all these

genes.

Our analysis

was made on the basis

from information about

resistance or susceptibil-

ity of winter wheat received from different authors by different techniques with different combinations of races in local

pathogen populations. Therefore, we consider the data on source of resistance and statistical estimations made by

comparing samples of resistant and susceptible cultivars as preliminary. Nevertheless, based on genealogical informa-

tion, the data will be useful in conditions of artificial inoculation with certain races of the pathogen and the use of a

standard set of differentials.

References.

Babayants LT and Dubinina LA. 1990. A novel donor of wheat resistance to bunt (Tilletia carries (DC) Tul., T. laevis

Kuehn.) and its genetical background. Rus J Genet 26(12):2186-2190 (in Russian).

Babayants LT, Dubinina LA, and Yushchenko GM. 1999. Genetical background of resistance to common bunt (Tilletia

carries (DC) Tul.) for new wheat lines. Cytol Genet 33(6):25-30.

Krivchenko VI. 1984. Resistance to bunt for grain crops. Moscow:Kolos. 304 pp. (in Russian).

Manjunath A and Nadaf SK. 1983. A ready reckoner of expected F3 breeding behaviour useful in linkage studies.

Madras Agric J 70(6):360-365.

Martynov SP and Dobrotvorskaya TV. 2000. A study of genetic diversity in wheat using the Genetic Resources Infor-

mation and Analytic System GRIS. Rus J Genet 36:195-202.

Novokhatka VG, Mochalova LI, and Odintsova IG. 1990. New genes in wheat for resistance to common and dwarf

bunt (Tilletia carries (DC) Tul., T. laevis Kuehn., T. controversa Kuehn.). Rus J Genet 26(10):1808-1814 (in

Russian).

Wilcoxson RD and Saari EE eds. 1996. Bunt and smut diseases of wheat: Concepts and methods of disease manage-

ment. CIMMYT, Mexico. 66 pp.

Table 4. Average contribution of hypothetical sources of resistance to common bunt for

Russian and Ukrainian winter wheat cultivars in groups of resistant and susceptible

accessions. Values are followed by letters that indicate significant differences at P < 0.05

by Duncan’s multiple range test.

Cultivars bred Cultivars bred

north of 49°N latitude south of 49°N latitude

Ancestor Genes Resistant Susceptible Resistant Susceptible

Agropyron glaucum BtZ 2.11 b0.42 a0.00 a0.02 a

Eliseevskaya (rye) 1.95 b0.35 a0.09 a0.32 ab

Yaroslav emmer 0.27 a0.17 a0.86 a0.89 a

Tr. timopheevii 0.24 a0.28 a0.98 a0.69 a

Petkus (rye) 0.00 a0.13 a0.17 a0.09 a

LV- Odessa (via Zemka) 0.45 a2.07 b12.05 d7.05 c

Oro Bt4, 7 0.22 a0.47 a0.90 a0.59 a

Florence Bt3 0.18 a0.38 a1.01 a0.55 a

Federation Bt7 0.19 a0.46 a0.82 a0.49 a

Hussar Bt1, 2, 5 0.11 a0.11 a0.35 a0.33 a

133

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Fig. 1. The tracing of sources of resistance to common bunt for line Selection 101 by GRIS. The resistant cultivars are

marked with a symbol , susceptible , and carriers of Bt-genes .

134

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

ITEMS FROM THE REPUBLIC OF SOUTH AFRICA

SMALL GRAIN INSTITUTE

Private Bag X 29, Bethlehem 9700, South Africa.

Plant Breeding – winter and facultative breeding program.

J.C. Aucamp, D.J. Exley, and H.A. Smit.

During 2002 Small Grain Institute released a new bread wheat cultivar named Komati. Komati is a facultative cultivar

with moderate vernalization requirements. The lodging resistance of this tall cultivar is good. Komati has a long

coleoptile (± 9.3 cm) and excellent resistance to preharvest sprouting, which has been confirmed successfully. Another

advantage is the high level of resistance against RWA infestations. Komati is susceptible to stripe rust. Though suscep-

tible, the application of a fungicide is only necessary when conditions are optimal for disease development. The cultivar

has no aluminium tolerance and planting on soils with low pH and high levels of acidification is not recommendable.

Komati is suitable for planting on low to high potential fields. Yields are stable and competitive with the best cultivars

available. Komati has an excellent hectoliter mass and falling number and good protein characteristics and, thus,

produces grain of a supreme grade. The cultivar meets with the set standards for flour extraction, protein quality, water

absorption, and mixing quality required by the milling and baking industries.

Plant Breeding – spring wheat irrigation program.

W.H.P. Boshoff and H.A. Smit.

The Wheat Technical Committee accepted BSP98/8 for final classification. The line will be marketed as Olifants.

Olifants yields above average, which appears stable over environments. Important agronomic characteristics of Olifants

are a medium growth period, good tillering, and strong resistance to lodging. The cultivar has excellent quality charac-

teristics that comply with requirements of the milling and baking industry. Olifants has high levels of resistance to local

foliar diseases including the currently prevailing pathotypes of stripe rust.

Genetic diversity.

T. van A. Bredenkamp and M.V. van Wyk.

The success of a breeding program depends mainly on the genetic diversity available. A constant need exists for the

incorporation of new germ plasm to improve locally adapted lines. Activities of the Small Grain Institute Germplasm

Collection consist of the conservation of small grain crops, namely wheat, barley, oats, triticale, and rye. This collection

is maintained in a cold room facility with a mobile shelving system for medium-term viability at ± 4°C.

International collaboration is of extreme importance because no breeding program can function effectively

without sufficient heritable diversity. Over the years a working relationship has been established between South Africa

and CIMMYT (Mexico), ICARDA (Turkey), Uruguay, and other countries. Wheat, barley, and triticale nurseries and

trials, segregating material, and interspecific crosses are imported annually. These lines are evaluated under quarantine.

Microenvironments conductive to disease development are created artificially ensuring high selection pressure. The

three quarantine sites are Bethlehem in the Free State, Riviersonderend in the Western Cape, and Vaalharts in the

Northern Cape.

135

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Doubled-haploid program.

A.F. Malan and H.A. Smit.

During the 2002 wheat season, several combinations were handled in the DH program. The main purpose of the

program is to enhance the development of pure breeding material of promising combinations identified in the different

breeding programs. The DH process is involved in the spring, winter, and prebreeding programs and assists in develop-

ment of lines with good disease resistance, superior quality aspects, and minor gene characteristics.

In the spring wheat-breeding program, cross combinations were made with special emphasis on stripe rust

resistance. A potential winter breeding lines’ progeny will be tested for agronomic traits, bread-making quality, and

disease resistance. Material from the prebreeding program includes combinations for RWA and leaf rust resistance. All

these DH lines will be tested in extensive field trials during the 2003 season.

Applying molecular and tissue-culture techniques to problems in disease resistance of wheat with

an emphasis on stripe rust.

R. Prins, V.P. Ramburan, and W.H.P. Boshoff (ARC-Small Grain Institute, RSA); L.A. Boyd (Department of Disease and

Stress Biology, John Innes Centre, UK); Z.A. Pretorius (Plant Pathology Department, University of Free State, RSA);

and J.H. Louw (Genetics Department, University of Stellenbosch, RSA).

Adult-plant resistance to stripe rust in the South African wheat cultivar Kariega was assessed in a DH-mapping popula-

tion made from the F1 of a cross between Kariega and the susceptible cultivar Avocet S. A partial linkage map covering

all 21 chromosomes was developed with 208 DNA markers and four alternative loci.

Interesting features of the linkage map include the low polymorphism observed in the D genome and a region

showing segregation distortion on chromosome 4A. The Ltn and Sr26 genes also were mapped in this study. Two major

QTL, together explaining about 55 % of the variation in the trait, were identified on chromosomes 2B and 7D, whereas

minor QTL explaining about 14 % of the variation were identified on chromosomes 1A and 4A. The QTL on 7D appears

to correspond to Yr18, a gene for APR to stripe rust. Markers fairly close to the QTL have been identified and these may

be used to detect the presence of these QTL regions in marker-assisted selection. The APR to stripe rust of Kariega

appears to be controlled by major QTL, in combination with other minor QTL, which is characteristic of APR in general.

The DH population developed and the linkage map constructed are valuable resources for future genetic studies that may

include studying APR, plant-pathogen interactions, and the mapping of additional traits polymorphic in this population.

Previous field trials of genetic material derived from Cappelle Desprez (CD) and Palmiet confirmed the

effectiveness of Yr16 (APR) against the South African pathotypes (6E16- and 6E22-). We know that CD also carries a

T5BS–7BS translocation that is a complicating factor in studying Yr16. Chromosome 2D SSR markers, previously

thought to be associated with Yr16, were tested on various resistant and susceptible lines. The molecular data suggest

that the position of Yr16 on chromosome 2D needs further verification. Various resistant plants were used in backcrosses

to Avocet S and Palmiet and the resulting F1s were used to produce DHs to simplify future genetic studies. These DH

lines will be evaluated for their stripe rust phenotypes in a field trial in 2003.

Preharvest sprouting and falling number.

A. Barnard.

The South African wheat-producing areas, especially the Eastern Free State, are highly subject to the risk of preharvest

sprouting because of summer rainfall that occurs just prior to or during harvest. Because preharvest sprouting is closely

related to falling number (FN), a substantial amount of research is done on both topics.

Thousands of wheat spikes obtained from various commercial and newly released cultivars are evaluated for

preharvest sprouting tolerance with the help of a rain simulator. This information is handed down to the commercial

farmer to enable him to make the right decision regarding his cultivar choice for the coming season. Recently, more

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

attention also was given to breeding programs for sprouting resistance with the help of the rain simulator and protein

electrophoresis. This technique is still in developmental and its usefulness still uncertain. Should this technique prove to

be useful, direct crosses can be made and the progeny screened for the presence of the necessary electrophoretic bands,

ensuring that, as sprouting resistance is a polygenic trade, none of the genes will be lost during the breeding program.

Since the incorporation of the FN method within the grading regulations, attention has been given to the

possibility of managing FN within a wheat production system. The effect of early termination of kernel development

(early harvest) on the FN of wheat and the effect of fertilizer on FN are being investigated.

Wheat production in the Summer Rainfall Region.

Because of the importance of cultivar choice in the Summer Rainfall Region, an extensive cultivar-evaluation program is

followed for each of these areas. Different cultivars are planted in each region and these cultivars are evaluated and

characterized in terms of yield reaction and stability in the different areas. Other characteristics that also are evaluated in

this program include important quality specifications such as hectoliter mass, protein content, and falling number. These

characteristics are used in recommending cultivars best suited for each area in the region.

Dryland production. Almost half of the South African wheat production is in cultivation under dryland conditions in

the Summer Rainfall Region. Because of the large variation in climatic conditions and soil types existing in this region,

wheat production is very challenging. Not only are good cultivation and management practices essential for successful

wheat production, but also the correct cultivar choice. The dryland production area is divided primarily into four

homogenous areas where different cultivars, mainly winter and intermediate types, are planted. All cultivar-evaluation

trials planted at 18 sites throughout the Western, Central, and Eastern Free State were successful and reported. Eighteen

entries were included in the trials, seven were from Small Grain Institute, six from Monsanto, and five from PANNAR.

Production under irrigation. Wheat produced under irrigation amounts to about 20 % of the total wheat production of

South Africa and has a stabilizing influence on the total production. Currently, six major irrigation regions exist,

although irrigation farming is expanding into new regions.

Mainly spring wheat cultivars are planted in a total of 44 evaluation trials at 23 localities in the different

irrigation areas. Entries in these trials originated from Small Grain Institute (7) and from Monsanto (4). A durum

cultivar also was included. ANOVA, AMMI analysis, and biplots are used in the interpretation of results and identifying

cultivar adaptation and stability in the different production regions. Results from these trials are available in a detailed

report.

Wheat production in the Winter Rainfall Region.

There are mainly two wheat producing areas in the Winter Rainfall Region:

•The Swartland area stretches from Durbanville in the south to the Sandveld area around Elandsbaai in the

north and from Saldana Bay in the west to the mountain ranges in the east.

• The Rûens or South Coast area, stretches from Botrivier in the west to the Albertina-district in the east and

from Aghullas in the south to the Langeberg mountain range north of Greyton through to Riversdal.

Spring wheat cultivars are grown in these two regions. These cultivars do not require the same amount of cold

to break their dormancy as that of the winter wheats grown in the rest of South Africa. Cultivar choice in the Winter

Rainfall Region is of extreme importance because of the varied climatic differences between cultivation areas. The

yields of available cultivars differ relative to the changing yield-potential conditions that exist in the Winter Rainfall

Region. Other important factors that also need consideration are grain quality, hectoliter mass, and disease susceptibility.

In the Winter Rainfall Region, the cultivar-evaluation program is run jointly by The Small Grain Institute and

The Directorate of Agriculture of the Western Cape. The program consists of 13 sites in the Swartland and 13 sites in the

Rûens, with 11 cultivars included in the trials. Cultivars, from ARC-Small Grain Institute, Monsanto, and PANNAR, are

annually tested for yield potential, quality, disease resistance, and adaptability.

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Karnal bunt in South Africa.

K. Naudé.

Karnal bunt was identified for the first time in South Africa in December 2000 in the Douglas irrigation area. Karnal

bunt is caused by the quarantine organism, Tilletia indica, and according to South African regulations, the occurrence

thereof should be reported to the National Department of Agriculture (NDA).

To date, the South African wheat industry has been protected against wheat imports from countries where KB

already occurs. After the identification of KB in South Africa, a KB Task Team was founded with the objective to

compile protocols to limit the spread of the disease in South Africa. These protocols include the testing of all registered

seed units and all commercial grain for the presence of teliospores produced by the fungus. Using quarantine regulations

and permits for the transportation of grain to intake points and mills also are included.

Karnal bunt occurrence in South Africa. As was the case with the 2001–02 wheat-production season, official surveys

were made by the NDA–Directorate Plant Health and Quality (NDA–DPHQ) to test seed and grain for the presence of

KB spores and infected kernels. All seed units of the 2001–02 season tested free of spores and infected kernels. Karnal

bunt spores and infected kernels, however, were found in grain from the Douglas and Koffiefontein areas. Infection also

was found on four farms in the Douglas district, and these farms were placed under quarantine. Results of the 2002–03

season will be available at a later date.

Control of Karnal bunt. At this stage wheat producers are making use of seed free of KB spores. The treatment or

nontreatment of seed with chemical fungicidal seed dressing is done at the producer’s own discretion. In areas where

KB has been identified, spraying twice with Triticonazole is recommended. A first application is done at 25 % ear

emergence, followed by a second application 10 days later. This spraying system is used by most wheat producers in the

Douglas area with the purpose of limiting KB infection to levels lower than 2 %.

The role of ARC–Small Grain Institute (ARC–SGI). The latest information regarding KB and its control is trans-

ferred to producers, agents, and advisors at farmers’ days and during courses on a continual basis. ARC–SGI tests all its

seed and grain at the KB Laboratory at Bethlehem. All ARC–SGI seed required for planting at the more than 90 locali-

ties country wide is washed at the KB Washing Facility according to procedures used by CIMMYT. Fifty lines and

cultivars of ARC–SGI were evaluated by CIMMYT in the 2001–02 season for KB resistance. These lines and cultivars

will be evaluated by CIMMYT again during the 2002–03 season. Planting of less susceptible or resistant cultivars in the

affected areas is regarded as the only sustainable solution for the control of KB.

Research into the utilization of nitrogen by wheat cultivars under irrigation.

W.M. Otto.

The objective of this research was to measure the yield and protein response of irrigated wheat cultivars to nitrogen (N)

management options. Furthermore, the effect of split N applications combined with residual soil mineral N on grain

yield and quality also is determined. The contributions of soil mineral N, plant uptake of N, and biomass development to

N management of the crop also were calculated. The aim is to develop an N management system that the producer can

implement to optimize all the relevant production factors.

The yield and grain protein responses to split applications of N applied at planting, late tillering, and flag-leaf

stages of six commercially available wheat cultivars (SST 876, SST 822, Kariega, Olifants, Baviaans, and Steenbras)

were measured. The trials were planted at Riet River, Vaalharts, Loskop, and Bethlehem.

The tested cultivars differed in response to split applications of applied N, with the magnitude of yield and

protein response linked to the adaptability and growth period of the respective cultivar. Measured residual soil mineral N

influenced yield response to N. A high level of soil mineral N (205 kg N/ha) decreased the response to applied N,

whereas where a low soil mineral N of 60 kg N/ha was measured, significant responses to applied N were found.

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Protein percentage of the grain increased with an application of 40 kg N/ha at the flag-leaf stage. A decrease in

yield of all the tested cultivars was found when the total N rate was applied at planting. The split application of N, where

80–120 kg N/ha applied at planting was followed by 40–80 kg N/ha at late tillering and 40 kg N/ha at the flag-leaf stage,

resulted in the highest yields and protein percentage of the grain. Plant N analysis at the measured growth stages

indicated that the split application of N increased plant N concentration to within the optimal N analysis range, showing

the potential use of this measurement in N management of the crop.

Wheat Quality Laboratory.

A. Barnard, C.W. Miles, K.B. Majola, M.L.T. Moloi, M.M. Raderbe, N.E.M. Mtjale, C.N. Matla, M.M. Mofokeng, M.L.

Dhlamini, and N.M. Mtshali.

One of the main objectives of the Quality Laboratory is to maintain a cost-effective, highly scientific, and objective

quality assessment of Small Grain Institute breeding lines, to incorporate contract work for milling and baking industries

and private companies, and to provide an objective service to wheat producers. To ensure accurate data to researchers

and external parties, the laboratory takes part in quarterly and monthly ring tests. A total of 57,410 analyses were

performed during 2002.

Soil Analyses Laboratory.

L. Visser.

Soil analyses form an essential part of a producer’s success. The laboratory provides this service and plant and water

analyses to external clients and researchers.

During the 2001–02 financial year, the laboratory performed 111,032 tests on 9,329 samples. Fifty-four percent

of these samples were received from external clients such as producers, advisors, and representatives of different

fertilizer companies. During December 2001, the laboratory bought a new Inductive Coupled Plasma Emission Spec-

trometer. The instrument is known for accurate, reliable, and fast results. With this instrument the laboratory can handle

a larger amount of samples/year and also analyze for elements such as sulphur and boron.

The laboratory is also involved in a research project to evaluate the soil fertility status of resource poor areas

where Small Grain Institute operates. The database will help identify trends like increases in soil acidity and also

improve the quality of technology transfer in future.

In order to ensure an accurate and reliable service to all clients, the laboratory runs internal control samples and

also belongs to Agri-LASA, a national control scheme.

During the past year the external income of the laboratory increased by 10 %. The main objective of the

laboratory is to improve on this by rendering an accurate and efficient service to all clients.

Personnel.

Ms. Vicki Tolmay was appointed program manager of Plant Protection replacing Dr. Hugo Smit. Ms. Anri Barnard has

resigned to pursue household duties. Labious Masike replaced Godwin Khorommbi as a researcher of Plant Protection.

Sanesh Raburam joined Crop Science as a research technician. Willem Otto was transferred to Crop Science to handle

the Cultivar Evaluation Programme under irrigation. Pieter Craven and Danie van Niekerk resigned from Small Grain

Institute.

Publications.

Boshoff WHP and Van Niekerk BD. 2002. Resistance in South African and foreign wheat cultivars to pathotypes

6E16A- and 6E22A- of Puccinia striiformis f. sp. tritici. So Afr J Plant Soil 19(1):27-36.

139

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Boshoff WHP, Pretorius ZA, and Van Niekerk BD. 2002. Establishment, distribution and pathogenicity of Puccinia

striiformis f.sp. tritici in South Africa. Plant Dis 86(5):485-492.

De Villiers BL. 2002. Glyphosate absorption and activity may be explained by the physical appearance of the spray

droplet residuals. In: Proc 17th So Afr Weed Sci Soc Cong, 18–21 June 2002, Umhlanga Rocks, South Africa.

Du Preez CC, Kotze E, and Steyn JT. 2001. Influence of wheat residue management practices on potassium distribution

in a Plinthosol. In: Proc 12th World Fertilizer Cong, 3–9 August, 2001, Beijing, China.

Du Preez CC, Steyn JT, and Kotze E. 2001. Long-term effects of wheat residue management practices on some fertility

indicators of an Avalon soil. In: Proc 23rd Natl Cong Soil Sci Soc So Afr, Pretoria, South Africa.

Hatting JL and Kaya HK. 2001. Entomopathogenic nematodes: prospects for biological control in South Africa. In:

Proc 13th Cong So Afr Ent Soc, 2–5 July 2001, Pietermaritzburg, South Africa.

Labuschagne MT, Mamuya IN, and Koekemoer FP. 2002. Canonical variate analysis of breadmaking quality character-

istics in irrigated spring wheat (Triticum aestivum). Cereal Res Commun 30(1-2):195-201.

Maré, R and Tolmay, VL. 2001. Why does annual Russian wheat aphid infestation on wheat fluctuate? In: Proc 13th

Cong So Afr Ent Soc, 2–5 July 2001, Pietermaritzburg, South Africa (Poster presentation).

Ntushelo K and Van Niekerk BD. 2002. Reaction of South African spring wheat cultivars to head blight caused by

Fusarium graminearum. So Afr J Plant Soil 19(1):50-51.

Otto WM and Kilian WH. 2001. Response of soil phosphorus content, growth and yield of wheat to long-term phospho-

rus fertilization in a conventional cropping system. Nutrient Cycling in Agroecosystems 61(3):283-292.

Prinsloo GJ. 2001. Can the parasitoid Aphelinus hordei (Kurdjumov) smell aphid infested wheat plants? In: Proc 13th

Cong So Afr Ent Soc, 2–5 July 2001, Pietermaritzburg, South Africa.

Tolmay VL. 2002. Resistance to biotic and abiotic stress in the Triticeae. Hereditas 135(2-3):239-242.

Tolmay VL. 2002. Economic and social aspects of pest management: pest tolerance in crops. In: Encyc Pest Manage-

ment, Marcel Dekker Inc, New York. Available online at http://www.dekker.com, 3 pp.

Tolmay VL and Maré R. 2001. Mechanisms of resistance to Russian wheat aphid (Diuraphis noxia) in resistant South

African wheat cultivars. In: Proc 13th Cong So Afr Ent Soc, 2–5 July 2001, Pietermaritzburg, South Africa.

Tolmay VL. 2001. Resistance is the key to a successful revolution. In: Proc 4th Internat Triticeae Symp, 10–12

September, 2001, Cordoba, Spain (Invited Keynote speaker for Session D Resistance to Biotic and Abiotic stresses).

UNIVERSITY OF STELLENBOSCH

Department of Genetics, Private Bag X1, Matieland 7602, South Africa.

G.F. Marais, H.S. Roux, A.S. Marais and W.C. Botes.

Triticale breeding.

Three new triticale cultivars will be available for commercial production in 2003, these are Bacchus, a selection from

CIMMYTs 28th ITYN-48 (SUPI 3//HARE 7265/YOGUI 1); Tobie, a selection from the local cross KIEWIET/4/

W.TCL83/HOHI//RHINO 4/3/ARDI 1; and Ibis, also derived from a local cross FLORIDA 201/17th ITSN 238 (=

DURUM WHEAT/BALBO//BOK”S”). Tobie is a very early maturing triticale with high grain yield and excellent

hectoliter mass, whereas Bacchus is a high-yielding, later maturing cultivar. Ibis is a late-maturing, tall straw cultivar

selected specifically for the production of fodder. Two cultivars released at Stellenbosch, Tobie and USGEN19 (late

maturing), also have been released in Ethiopia under the names Sinan and Maynet, respectively, as part of a collaboration

with the Ethiopian Bureau of Agriculture, Amhara Region, and the German G.T.Z. (Technische Zusammenarbeit)

program coördinated by Dr. K. Feldner.

Recurrent selection of wheat.

A recurrent-selection procedure for wheat based on genetic male sterility and hydroponic culture of cut tillers was

continued and further improvements of the technique were made (Fig. 1). The breeding cycle could be reduced to 4

years (1 year for making crosses and 3 years for inbreeding and field evaluating the F4 and F6 male parents). Single-seed

descent steps were introduced to advance from the F1 to the F6 in the course of 2 years. To accomplish this, two addi-

tional plantings (F2 and F3) are made during the summer months in an uncooled greenhouse. The F4 is again planted in

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Fig. 1. Recurrent mass selection scheme for spring wheat.

Reserve

• Select for leaf and stem rust

F1 hybrid seed seedling resistance

• Marker-aided selection ?

Pollinate selected F1 1/2 Msms 1/2 msms

female plants with (Male sterile) (Male fertile)

selected F6 male plants

Harvest 5–10 random F2 seeds/plant

(800–1,000 plants). Plant (single pot)

in greenhouse by 15 October, year 1

(spring).

Harvest single and bulked seeds of each

pot in December, season 1. Make SDS-

sedimentation tests. Plant 5–10 random

F3 seeds/selection in January of year 2

(summer greenhouse).

Plant F4 rows in the field (normal winter

planting), select single plants from superior

rows.

Plant F5 rows in summer (irrigation).

Inoculate and screen for adult-plant

stem rust resistance.

Evaluate as unreplicated F6 plots; micro-mill,

and evaluate mixograph and kernel hardness.

Advanced trials: select for agrotype,

disease resistance, yield, and quality.

Regional tests

Select for resistance

genes and/or

markers.

Year 4 Year 3 Year 2 Year 1

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

the normal growing season (winter) and the F5 planted under irrigation during the summer. The F6 is used for the first

yield trials (plots) in the third winter. This modification has several advantages. The shorter breeding cycle allows for a

more rapid increase in the frequency of desirable alleles of disease resistance loci. However, properly selecting for

agronomic and quality characteristics of lower heritability is still possible. Inbreeding to the F6 has the same effect as the

use of DH technology but can be achieved at considerably reduced costs. When inbred male parents are selected for

marker or disease-resistance loci, very rapid shifts in the frequency of desirable alleles occur. Once the frequency of the

favorable allele of a number of genes has been raised to 0.70 or higher, a significant proportion of inbreds will have these

genes fixed in their genotypes, i.e., the procedure facilitates gene pyramiding. The advantage of pyramiding genes in

this manner is that there is no yield ceiling as is the case with backcross-based procedures and numerous diverse geno-

types with pyramided genes can be generated over time.

Genetic studies.

In 1993, we initiated a program for transferring leaf rust-resistance genes identified in a collection of wild Triticum

species. We have developed advanced material of 11 sources that show effective resistance to all known local patho-

types of one or more of the diseases leaf, stem, and stripe rusts. These include a subset of six lines in which resistance

(derived from T. turgidum subsp. dicoccoides, Ae. sharonensis, Ae. speltoides, Ae. peregrina, and Ae. kotschyi) appears to

occur on wheat chromosomes and five addition lines with added chromosomes from Ae. peregrina, Ae. umbellulata, Ae.

biuncialis, and Ae. neglecta. In several instances, promising stem rust and/or stripe rust resistance genes were co-

transferred with leaf rust resistance. The stripe rust resistance genes (from T. turgidum, Ae. sharonensis, Ae. speltoides,

Ae. peregrina, Ae. kotschyi, and Ae. biuncialis) also were effective against four Australian pathotypes and appeared to be

novel (evaluations done by Dr. Colin Wellings, University of Sydney). Leaf rust-resistance genes in 10 of the sources

showed promising resistance to commonly occurring Western Canadian pathotypes of the disease (evaluated by Dr.

Brent McCallum, Cereal Research Centre, Winnipeg, Canada). Stem rust-resistance genes from two Ae. speltoides

sources were tested with Western Canadian stem rust pathotypes (Dr. Thomas Fetch, Cereal Research Centre, Winnipeg,

Canada). One source showed resistance to all pathotypes whereas another was susceptible to one of the pathotypes. All

the genes appear to have a wide spectrum of resistance to justify continued introgression into wheat. Preliminary results

would suggest that the Ae. kotschyi-derived genes (leaf and stripe rust resistance) occur on chromosome 2D, whereas leaf

and stripe rust-resistance genes from Ae. sharonensis are on 3B. Resistance from Ae. biuncialis and Ae. neglecta appears

to be on group-3 chromosomes of these species. Some of the resistance genes have preferential transmission and the Ae.

speltoides-derived genes may involve gametocidal effects.

A unique, Th. distichum/ 4x rye hybrid (95M1) with genomes J1dJ2dRR allowed us to identify four Thinopyrum

chromosomes apparently involved with salt tolerance. When 95M1 was pollinated with diploid rye it yielded F1 off-

spring with primarily 21 chromosomes (two complete rye genomes and seven Thinopyrum chromosomes). Apparently,

the closely related homoeologous chromosomes of the J1d and J2d genomes regularly formed bivalents during megasporo-

genesis, and egg cells mostly received a random, yet balanced set of seven Thinopyrum chromosomes. F1 plants were

tested for salt tolerance and a set of 15 highly salt-tolerant F1 plants were selected and maintained as clones for several

years. These plants were C-banded and the Thinopyrum chromosomes in each line were determined. By comparing

segregation patterns, the Thinopyrum chromosomes were grouped into seven homoeologous pairs. For each of four

homoeologous pairs, one of its members occurred at a higher than expected frequency, implying that these chromosomes

are primarily being expressed under salt-stress conditions. The results could be confirmed by backcrossing two of the

most tolerant F1 plants to diploid rye. Although the critical chromosomes can be identified through C-banding, an

attempt also was made to find an RFLP marker for each. RFLP probes, diagnostic for the group 2, 3, 4, and 5

homoeologues of wheat, detected polymorphisms on the respective critical Thinopyrum chromosomes. However, the

preliminary allocation of the critical chromosomes to homoeology groups needs to be confirmed using more and varied

markers. An attempt also was initiated to develop triticale plants with disomic additions of the respective critical

Thinopyrum chromosomes. Disomic addition lines producing the group-3 and 5 RFLPs of two of the target chromo-

somes have been recovered and are being used in attempts to induce translocations to triticale chromosomes.

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ITEMS FROM SPAIN

UNIVERSIDAD POLITECNICA DE MADRID

Departamento de Biotecnologia, E.T.S. Ing. Agronomos.- C. Universitaria, 28040,

Madrid, Spain.

A. Delibes, I. López-Braña, M. J. Montes, and C. González-Belinchón.

UNIVERSIDAD DE LLEIDA

Institut de Recerca i Tecnologia Agroalimentaries (UdL-IRTA).

Rovira Roure, 191-25198 Lleida, Spain.

J.A. Martín-Sánchez, E. Sin, C. Martínez, and A. Michelena.

JUNTA DE EXTREMADURA. SERVICIO DE INVESTIGACION AGRARIA.

Departamento de Fitopatología. Ap. 22 CP. 06080 Badajoz, Spain.

J. del Moral, F. Pérez-Rojas, F. J. Espinal, and M. Senero.

Studies in relation to the Hessian fly-resistance gene (H30) transferred from the wild grass

Aegilops triuncialis to hexaploid wheat.

The transfer line TR-3531 (42 chromosomes), derived from the cross ‘T. turgidum/Ae. triuncialis//T. aestivum’ and

carrying the H. avenae resistance gene Cre7 (Romero et al. 1998), showed a high level of resistance to the M. destructor

biotype prevailing in southwestern Spain. A single, dominant gene (H30) determines the Hessian fly resistance in this

introgression line (Delibes et al. 2001), and its linkage with an isozyme marker (Acph-U1) has been studied. A phos-

phatase marker, resolved into two components, is present in the TR-3531 line, Ae. triuncialis (UC), Ae. umbellulata (U),

and the amphiploid Chinese Spring/Ae. umbellulata (ABDU), but is absent in Ae. caudata (C). Linkage between H30

and the Acph-U1 marker (associated with the U genome) was determined by analyzing 126 individual (TR-3531/H-10-

15) F2 plants. The kernels were cut transversely and the halves without embryos were used to obtain phosphatase

zymograms, an enzymatic system associated in wheat with homoeologous group 4 (Delibes et al. 1997a). The linkage in

this cross is not very tight, which would be consistent with the recombination expected of the ability of the C genome to

suppress the Ph-diploidization mechanism of wheat (Romero et al. 1998).

F2 progeny, derived from crosses between different wheat cultivars from Uniform Hessian Fly Nursery (UHFN

and H-93-33 transfer line) and with other sources of resistance, and TR-3531 were tested for resistance in field condi-

tions in order to determine if the new resistance gene was allelic with the H3, H5, H6, H12, H13, H18, H21, or H27

genes. Although the cultivars from UHFN are effective against Hessian fly in the United States, there is no evidence that

the selected genes confer resistance to biotype present in Azuaga (southwestern Spain). The results and summarized in

Table 1. All UHFN cultivars tested with different genes were resistant to this biotype, except the cultivar Abe with the

gene H5, which showed a inconsistent reaction. The resistance gene H30 in line TR-3531 is nonallelic with respect to

the genes H3, H6, H12, H13, H18, and H21 present in wheat cultivars from UHFN and H27 in the introgression line H-

93-33 (Delibes et al. 1997a and b). Previously, we demonstrated that H30 in TR-3531 line was not allelic with respect

H9 and H11 present in cultivars Ella and Kay, respectively (Delibes et al. 2001).

Advanced lines with the H30 gene were obtained by backcrossing the transfer line and different commercial

wheats (cultivars Anza, Betres, Cajeme, Cartaya, Marius, Rinconada, and Osona) as recurrent parents. In all advanced

lines, the infestation level was higher, but in the same range, than the donor. Several agronomic characteristics were

studied in 16 advanced lines and the results of three of the lines are summarized in the Table 2. The best results were

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

achieved with the Ma-6

line, which displayed

good agronomic charac-

teristics, in comparison to

the susceptible controls,

for the three traits studied.

Another fact that in-

creases the importance of

this line is that it also

carries the CCN resistance

gene Cre7.

Coöperation with other

institutions. We are

coöperating with Acorex

(Cooperative of

Extremadura farmers).

Financial support. This

work was supported by

grants AGL2001-3824-CO4, PTR 95-0496-OP CO1021001 from CICYT “Comision Interministerial de Ciencia y

Tecnologia” of Spain and IPR99A042 (Junta de Extremadura).

References.

Delibes A, Del Moral J, Martín-Sánchez JA, Mejías A, Gallego M, Casado D, Sin E and López-Braña I. 1997a. Hessian

fly-resistance gene transferred from chromosome 4Mv of Aegilops ventricosa to Triticum aestivum. Theor Appl Genet

94:858-864.

Delibes A, López-Braña I, Martín-Sanchez JA, Sin E, Martínez C, Michelena A, Del Moral J and Mejías A. 1997b.

Transfer of one gene for resistance to Hessian fly (Mayetiola destructor) from Aegilops ventricosa to cultivars of

wheat. Ann Wheat Newslet 43:214-215.

Table 1. Hessian fly reactions of the parents, F1, and F2 populations from crosses between wheats with

different resistance genes and the resistant line TR-3531, a carrier of the H30 gene. Hessian fly reaction of

line R-3531 is 25R:0S. R = resistance and S = susceptibility to Hessian fly. UHFN = Uniform Hessian Fly

Nursery.

Hessian fly reaction

Crosses between cultivars of the UHFN / TR-3531

UHFN No. F1No. F2 X2(1:d.f.)

line plants plants 15:1 ratio

Cultivar or Chromo-

UHFN line Gene some R:S R:S R:S Value Probability (P)

Howell H3 5A 18:0 7:0 154:8 0.46 0.5

Monon H3 5A 19:0 9:0 187:6 3.25 0.05<P<0.1

Caldwell H6 5A 29:0 8:1 218:14 0.02 0.9

841453

H15-1-1-2-5-2 H12 5A 26:1 2:0 95:1 4.45 0.01<P<0.05

86925 RA1-16 H13 6DL 18:2 3:0 140:7 0.55 0.3<P<0.5

Brule H18 —15:4 7:0 141:2 5.74 0.01<P<0.05

KS86HF012-23-6 H21 2BS 19:0 7:0 243:6 6.27 0.01<P<0.05

H-93-33 H27 4Mv21:0 7:0 50:5 0.76 0.3<P<0.5

Table 2. Agronomic characteristics of three advanced lines with the H30 resistance gene

in comparison to three bread wheat cultivars.

Yield Kernels 1,000-kernel

Cultivar or line (g/m2)/spikelet weight (g)

T. aestivum cv. Osona 1,240.57 2.71 29.49

T. aestivum cv. Astral 897.55 1.64 30.17

T. aestivum cv. Adalid 1,687.51 2.65 33.14

Ma6: TR/OS⊗//OS/3/RN/4/OS/5/RN/6/AZ 3⊗1,914.12 3.00 43.37

Ma4: TR/BT⊗//AL/3/MA/4/3*BT 2⊗1,336.08 2.84 27.65

Ma3: TR/3*OS//4*CYÄ/3/CJ 4⊗1,486.86 2.64 32.50

Least significant difference (LSDP<0.05) 409.16 0.47 5.05

Abbreviations used: Ma = advanced lines, TR = TR-353 line, AZ = Anza, BT = Betres, CJ =

Cajeme, CY = Cartaya, MA = Marius, OS = Osona, RN = Rinconada, ⊗ = selfing.

144

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Delibes A, López-Braña I, Montes MJ, Gómez-Colmenarejo M, Romero D, Andrés MF, Martín-Sánchez JA, Sin E,

Martínez C, Michelena A, del Moral J, and Mejías A. 2001. Transfer of resistance genes to Hessian fly and Cereal

Cyst nematode from Aegilops triuncialis to hexaploid wheat and its use in breeding programs. Ann Wheat Newslet

47:198-200.

Romero MD, Montes MJ, Sin E, López-Braña I, Duce A, Martín-Sánchez JA, Andrés MF and Delibes A. 1998. A cereal

cyst nematode (Heterodera avenae Woll.) resistance gene transferred from Aegilops triuncialis to hexaploid wheat.

Theor Appl Genet 96:1135-1140.

Publications.

Andrés MF, Romero D, Montes MJ and Delibes A. 2001. Genetic relationships and isozyme variability in Heterodera

avenae complex determined by isoelectrofocusing. Plant Pathol 50:270-279.

Andrés MF, Melillo MT, Delibes A, Romero MD and Bleve-Zacheo T. 2001. Changes in wheat root enzymes correlated

with resistance to cereal cyst nematodes. New Phytol 152:343-354.

Andrés MF, Melillo MT, Delibes A, Romero MD and Bleve-Zacheo T. 2001. Biochemical and cytochemical changes in

a resistant wheat/ Aegilops ventricosa introgression line during cereal cyst nematode infection. In: Proc 11th Cong

Mediter Phytopathological Union and 3rd Cong Sociedade Portuguesa de Fitopatologia, Evora (Portugal), 17–20

September.

Andrés MF, Melillo MT, Delibes A, Romero MD and Bleve-Zacheo T. 2002. Biochemical and cytochemical changes in

a resistant wheat/ Aegilops ventricosa introgression line during cereal cyst nematode infection. Phytopath Meditet (in

press).

Delibes A, López-Braña I, Montes MJ, Gómez-Colmenarejo M, Romero MD, Andrés MF, Martín-Sánchez JA, Sin E,

Martínez C, Michelena A, del Moral J and Mejías A. 2001. Transfer of resistance genes to Hessian fly and Cereal

Cyst nematode from Aegilops triuncialis to hexaploid wheat and its use in breeding programs. Ann Wheat Newslet

47:198-200.

Delibes A, López-Braña I, Montes MJ, Gómez-Colmenarejo M, González-Belinchón C, Romero D, Andrés MF, Martín-

Sánchez, JA, Sin, E, Martínez C, and Michelena A. 2002. Differential induction of defence-enzymes and chromo-

somal location of two Heterodera avenae resistance genes transferred to wheat from Aegilops ventricosa. Ann Wheat

Newslet 48:165-167.

Delibes A, López-Braña I, Montes MJ, Gómez-Colmenarejo M, González-Belinchón C, Martín-Sanchéz JA, Sin E,

Martínez C, and Michelena A, del Moral J, Pérez-Rojas F, and Espinal FJ. 2002. Resistance to Hessian fly conferred

by the gene H27. Relationships with other sources of resistance and its effect in some agronomic traits. Ann Wheat

Newslet 48:167-169.

Del Moral J, Delibes A, Martín JA, Pérez-Rojas F, Rubiales D, Merino M, Sin E, Martínez C, Montes MJ, Sereno M and

Campos L. 2001. Selección de una colección de líneas avanzadas de trigo con resistencia a Mayetiola destructor

Say. por su comportamiento frente a los patógenos y accidentes fisiológicos más importantes de este cultivo en la

Campiña Sur de Extremadura. In: Proc II Congr Nac Ent Aplicada, VIII Jornadas Científicas de la S.E.E.A.

Pamplona (Spain), November (in Spanish).

Del Moral, J, Delibes A, Martín-Sánchez JA, Mejías A, López-Braña I, Sin E, Montes MJ, Pérez-Rojas F, Espinal F and

Senero M. 2002. Obtención de líneas de trigo resistentes a Mayetiola destructor say en la campiña sur de

Extremadura. Boletín Sanidad Vegetal 28(4):585-590 (in Spanish).

Martín-Sánchez JA, Gómez-Colmenarejo M, Del Moral J, Sin E, Montes MJ, González-Belinchón C, López-Braña I,

Delibes A. 2002. A new Hessian fly resistance gene (H30) transferred from the wild grass Aegilops triuncialis to

hexaploid wheat. Theor Appl Genet (in press).

Martín-Sánchez JA, Montes MJ, López-Braña I, Romero MD, Sin E, Martínez C, Andrés MF, Gómez-Colmenarejo M,

González-Belinchón C, Delibes A. 2002. Differential induction of defence-enzymes and chromosomal location in

wheat/Aegilops ventricosa introgression lines of Cre2 and Cre5 Heterodera avenae resistance genes. In: Eucarpia

Cereal Sect Meet, Salsomaggiore, Italy, 21–25 November.

Montes MJ. 2001. Transferencia de un gen de resistencia al nematodo de quiste de los cereales, Heterodera avenae,

desde Aegilops triuncialis a trigo hexaploide. Relaciones con otros genes de resistencia. PhD Thesis. Universidad

Politecnica de Madrid, Spain (in Spanish).

Montes MJ, Gómez-Colmenarejo M, Andrés MF, Romero MD, López-Braña I, Delibes A. 2002. Inducción de enzimas

relacionados con el estrés oxidativo en líneas de introgresión trigo/Aegilops portadoras de los genes de resistencia a

Heterodera avenae Cre2, Cre5 y Cre7. In: Proc XI Congr Soc Esp Fitopatología, Almería, Spain, 14–18 October (in

Spanish).

145

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Montes MJ, López-Braña I, Gómez-Colmenarejo M and Delibes A. 2001. Enzimas relacionadas con el estres oxidativo

inducidas en raices de trigos resistentes Heterodera avenae, en respuesta a la infestacion por el nematodo. Congr Soc

Esp Genet, Sevilla, Spain, 19–22 September (in Spanish).

Montes MJ, López-Braña I, Romero, MD, Sin E, Andrés MF, Martín-Sánchez JA, Gómez-Colmenarejo M and Delibes

A. 2002. Comparative study of two Heterodera avenae resistance genes from Aegilops ventricosa. Differences in

defence-enzymes induction and chromosomal location in wheat/Ae. ventricosa introgression lines. In: Proc 4th

Internat Congr Nemat, Tenerife, Spain, 8–13 June.

Ogbonnaya S, Seah I, Delibes A, Jahier J, López-Braña I, Eastwood RF and Lagudah ES. 2001. Molecular-genetic

characterisation of nematode resistance from Aegilops ventricosa and its derivatives in wheat. Theor Appl Genet

102:263-269.

Sin E, Montes MJ, Romero MD, López-Braña I, Andrés MF, Martín-Sánchez JA, Benavente E, Gómez-Colmenarejo M,

Delibes A. 2001. Transferencia del gen de resistencia a Heterodera avenae (Cre7) desde Aegilops triuncialis a trigo

hexaploide. In: Congr Soc Esp Genet, Sevilla, Spain, 19–22 September (in Spanish).

ITEMS FROM TURKEY

CIMMYT AND THE MINISTRY OF AGRICULTURE AND RURAL AFFAIRS

P.K. 39 Emek, 06511 Ankara, Turkey.

H.J. Braun, A.R. Hede, J. Nicol, and B. Akin (CIMMYT–Turkey); M. Keser, N. Bolat, N. Colak, H. Ekiz, S. Taner, S.

Ceri, F. Partigoc, L. Cetin, S. Albustan, F. E. Donmez, Dusunceli, S. Yazar, I. Ozseven, I. Ozturk, and T. Yildirim

(Ministry of Agriculture and Rural Affairs); and M. Mousaad and A. Yahyaoui (ICARDA, Aleppo, Syria).

The International Winter Wheat Program (IWWIP) is a joint program carried out by the Ministry of Agriculture of

Turkey, CIMMYT, and ICARDA. The two main objectives of the program are to develop broadly adapted, disease-

resistant, high-yielding winter wheat germ plasm for the winter and facultative wheat-growing areas in Central and West

Asia and North Africa (CWANA) and to help facilitate germ plasm exchange among the winter wheat-breeding programs

around the world.

Of the 103 million ha of wheat grown in the least-developed countries. Approximately 31 x 106 ha are winter

and facultative wheat, of which 16.5 x 106 ha are sown in CWANA; 13 x 106 ha in China; and 1 x 106 ha in South

America, North Africa, and North Korea. After China, Turkey is the 2nd largest winter wheat grower among the least-

developed countries with 6.6 x 106 ha; followed by Iran with 4 x 106 ha.

News on germ plasm development and cultivar releases.

Since 1980, 27 cultivars from the IWWIP program have been released in CWANA. In 2002 alone, nine cultivars were

released in Afghanistan (1), Georgia (1), Turkey (6), and Uzbekistan (1) (Table 1). Three of these cultivars are targeted

for rainfed areas and five for irrigated/supplementary irrigation conditions. Thirty-four cultivars are presently included

in registration trials in Armenia (6), Georgia (1), Kazakhstan (2), Kyrghyzstan (7), Tajikistan (6), Turkey (5),

Turkmenistan (3), and Uzbekistan (4).

The winter wheat program draws heavily on the winter (W)/spring (S) crosses. A major contribution to the

winter wheat program is made through the spring wheat lines developed at CIMMYT–Mexico, which are crossed with

winter wheats. Many of the most successful CIMMYT spring wheats were derived from W/S crosses. Now, the same is

happening for winter wheat. More than 75 % of the IWWIP lines released or in registration trials are selected from

crosses between winter and spring wheat lines and three-way crosses (winter/spring//winter). These WSW-derived

cultivars are now making their way into registration trials throughout the CWANA region (Fig. 1).

146

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

News on germ plasm exchange:

the case of yellow rust.

The Facultative and Winter Wheat

Observation Nursery (FAWWON) has

served as the main vehicle for facilitating

germ plasm exchange among winter

wheat programs. This nursery consists of

lines developed by the IWWIP program

and of cultivars submitted by national

programs, university programs, or private

companies from countries in CWANA,

western and eastern Europe, China,

South America, and the U.S.A. The 11th

FAWWON consisting of 146 entries was

distributed for planting in the 2001–02

cropping cycle to around 80 coöperators

from more than 40 countries

Yellow rust is one of the most important

leaf diseases for the winter wheat areas in west

and central Asia. Within the last decade, CWANA

countries suffered several major yellow rust

epidemics, with losses up to 50 %. Fig. 2 shows

the maximum yellow rust score from the evalua-

tion of the 11th FAWWON across 10 locations in

Iran (5), Turkey (1), Azerbaijan (1), Tajikistan (1),

Syria (1), and China (1). Characteristically, most

lines developed by the IWWIP program show

good levels of resistance, whereas most other lines

are highly susceptible to yellow rust. The fact that

many yellow rust-susceptible, but otherwise

excellent, lines with highly favorable characteris-

tics will be dismissed by breeders due to yellow

rust susceptibility and, therefore, not utilized by

breeding programs has forced us to think of ways

to restructuring the FAWWON nursery. These

changes will be implemented within the coming

year.

Table 1. International Winter Wheat Program-derived wheat cultivars registered in Central and West Asia

and Northern Africa in 2002.

Country Cultivar Cross Type

Afghanistan Solh02 OK82282//BOW/NKT WS

Georgia Mtsjetslaua 1 TAST/SPRW//ZAR WS

Turkey Soyer ATAY/Galvez WS

Turkey Yildirim ID800994.W/VEE WS

Turkey Daphan JUP/4/CLLF/3II14.53/ODIN//CI14431/WA00477 WS

Turkey Bagco 2002 HN7/Oorfen//BJN8/3/SERI82/4/74CB462/Trapper//Vona WW

Turkey Nenehatun ND/P101/Blueboy

Turkey Sakin PI/FUNO*2//VLD/3/CO723595 WW

Uzbekistan Dostlik YMH/TOB//MCD/3/LIRA WS

Fig 1. Frequency of winter (W)/spring (S), W/W, and W/S//W crosses in

International Winter Wheat Program-derived cultivars registered since

1980 or lines presently in registration trials in Central and West Asia and

Northern Africa.

Fig 2. Maximum scores for resistance to yellow rust across 10

locations in six countries in Central and West Asia and North

Africa of 146 entries in the 11th Facultative and Winter Wheat

Observation Nursery. Entries within each group of origin are

sorted by increasing susceptibility.

147

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Research on root rots and nematodes.

H. Aktas, A. Bagci, N. Bolat, O. Buyuk, C. Cekic, H. Ekiz, I. Gultekin, H. Hekimhan, Y. Kaya, M. Keser, I. Ozseven, M.

Tekeoglu, H. Toktay, B. Tunali, A. Tulek, Zafer Uçkun, A.F. Yildirim, and A. Yorgancilar (Turkish Ministry of Agricul-

ture and Rural Affairs); H.J. Braun, M. Mergoum, J. Nicol, R. Trethowan, M. van Ginkel, and M William (CIMMYT

International); H. Elekcioglu (Cukurova University); E. Sahin (Osman Gazi University); and R. Rivoal (INRA/ENSAR

France).

Since 1998, the Ministry of Agriculture and Rural Affairs in Turkey (MARA) in collaboration with CIMMYT staff based

in Turkey have initiated two key National/International projects. One of these is on cereal nematodes and the other on

cereal root rots. These projects cover a range of research areas including;

– surveys,

– economic importance and population dynamics,

– identifying of sources of resistance, and

– control methods emphasising plant genetic resistance.

Below is a brief summary of some of our findings to date. We very much encourage anyone interested in

collaborating with our program to make contact with us.

Preliminary surveys. These surveys have been conducted on the Central Anatolian Plateau, the major winter wheat-

growing region of Turkey. The objective was to understand the distribution of two economically important cereal

nematodes, cyst (Heterodera spp.), and lesion (Pratylenchus spp.). Seventy-two percent of the root samples and 83 % of

the soil samples contained cysts and in approximately 40 % of soil samples one or both lesion nematodes were found.

Cereal cyst nematode was identified to species level using both traditional morphology and a RFLP PCR-based molecu-

lar method. None of the samples contained H. avenae, the most common cereal cyst nematode documented. Instead, 40

% of the samples contained H. latipons, 32 % H. filipjevi, and 28 % a mix a both species

A range of Fusarium species have been isolated from cereal crown roots with the most frequently isolated

species being F. culmorum, F. nivale, F. psuedograminearum, F. acuminatum, and F. heterosporum.

The taxonomy of cyst nematode is very time consuming and difficult, and work is underway to optimize the

molecular technology to identify the different species of Heterodera. We hope in the near future to relate and collate

survey data with both classical morphology and molecular methods.

In many cases, several species of nematodes and root rots are present in the same soil, suggesting that we are

dealing with a root disease complex and management strategies need to account for this. In addition, zinc deficient soils

are widespread and can be considered part of the problem complex (Cakmak et al. 1999).

To understand the economic importance and population dynamics of both nematodes and root rots, multi-

location and multi-year yield trials are being conducted. Work with root rots is more advanced than with nematodes.

Data from 2-year yield trials with and without root rot inoculation (inoculated as a mix of both Fusarium species and

Bipolaris) indicate that most of the common winter wheat cultivars grown in Turkey are intolerant suffering average

yield losses of 37 %. Furthermore, preliminary data of only 1 year suggests strongly that sources of spring wheat

resistance identified in Australia and confirmed at CIMMYT–Mexico also are resistant and have high tolerance under

field conditions in Turkey. We are in the second year of field testing with nematodes in three locations. Preliminary data

from last year clearly show economic grain loss, with most common winter wheat cultivars being intolerant and suscep-

tible (i.e., allow nematode multiplication) to both the lesion and cyst nematodes. Detailed knowledge is available about

the population dynamics of the lesion nematodes (P. thornei and P. neglectus) and at least one of the species of cyst

nematode (H. avenae). However, little is known about the biology and behavior of other Heterodera species that are

commonly found in winter wheat areas of the world, namely H. filipjevi and H. latipons. In addition to monitoring field-

population dynamics, we also conduct a range of basic biological experiments to identify factors that affect the hatch of

the cysts and to understand the duration of the life cycle and the infection process.

Identification of sources of resistance. A major focus of the work is to screen winter wheat and identify sources of

resistance to both the key nematodes and root rots. Again, this work is more advanced with root rots than nematodes on

winter wheat. Over 100 crosses/year are being made with sources of both nematode and root rot-resistant germ plasm

148

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

(Nicol 2002; Nicol et al. 2001). Spring wheat lines with confirmed root disease resistance also are used in the crossing

program.

A large screening program in the south of Turkey near Konya was established where annually around 2,000

accessions are inoculated with a root rot complex. The best accessions form screening nurseries are further tested in

replicated screening and yield trials. Resistant lines are confirmed based on field trials and greenhouse work. Screening

for nematode resistance concentrates on field screening at present, but work is underway to establish pure nematode

cultures in the laboratory and ultimately screen with individual nematodes under controlled laboratory conditions. With

cyst nematode, more work is required to understand the biology before such screening can be conducted on a larger

scale.

Work within CIMMYT also is utilizing the tools of molecular biology in a MAS strategy. Several markers for

known nematode-resistance genes developed in Australia are optimized and are being used routinely on CIMMYT germ

plasm. However, given the complexity of the nematode in the region we need to confirm the effectiveness of these

known sources of resistance with the range of nematode populations from the region.

Once we have more advanced plant populations (F4/F5) where resistance has been incorporated, we will conduct

confirmation screening to validate the incorporation of resistances using both traditional and molecular tools where

appropriate. As we produce these root disease resistant wheat lines, they will be distributed through the international

nurseries.

Cereal nematodes and root rots can be controlled in several ways. The major emphasis in our program is placed

on using plant genetic resistance. Because resistance alone is probably not the complete answer (as many resistances are

partial), other methods need to be investigated. To control root rots, we will look at the effects of seed treatment with

fungicides, application of microelements (including B, Cu, Fe, Mn, S, and Zn), seed-sowing density, and rotation

experiments of cereals with other non-cereal crops (such as canola and sugar beet). With nematodes, we will look at the

effect of crop rotation and management practices (such a conservation tillage and cultivation) on nematode numbers. As

has been proven in the U.S. and Australia, there is no doubt that agronomic practices have a key role to play in the

control of these pathogens.

Progress to date on breeding for root rot resistance and tolerance.

Extensive field screening over the last 3 years has assessed the resistance of winter wheat germ plasm against root rots

under field conditions. Resistance is defined as a reduction in symptom development of the disease. In Cumra, 40 km

south of Konya, field-observation plots were assessed by inoculating seed with a combination of root rot species

(Fusarium and Bipolaris) and comparing symptom development against uninoculated plots. These lines are now

entering yield trials to assess tolerance (yield loss) and also have been extensively crossed in the IWWIP program. The

best entries after 3-year screening are shown in Table 2. Several of the identified lines are widely grown cultivars such

as Gerek 79, Dagdas, and Katia-1.

Training. The IWWIP program is training Turkish scientists and scientists from the region in the field of soil disease

cereal research. This includes postgraduate training and special courses such as the one planned for June 2003 in Turkey.

Several Australian pathologists will attend this course to provide their expertise and knowledge. The training course is

called ‘Soil Borne Pathogens of Cereals’ and will be from 14–28 June under the coördination of the IWWIP. Participants

will be trained to work with both nematodes and root rots. We are very grateful to the sponsors, principally led by the

ATSE Crawford Fund, CIMMYT, MARA, ICARDA, GRDC, ACIAR, and the Kirkhouse Trust.

Concluding remarks. We believe by conducting this highly focused, complex and difficult research we can clearly

define the soilborne constraints in the winter wheat regions of CWANA and ultimately significantly improve wheat

production and sustainability of the cropping systems in our region. The key to this will involve a breeding approach to

produce high-yielding, quality-adapted germ plasm combined with multiple root disease resistances and microelement

efficiencies, complimented with appropriate management practices. This work is large and encompassing, and we

welcome any collaboration from interested parties.

149

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Publications.

Nicol JM. 2002. Important Nematode Pests of Cereals. In: Bread Wheat Production and Improvement, FAO Plant

Production and Protection Series, FAO, Rome, Italy. Pp. 345-366.

Nicol JM, Rivoal R, Bolat N, Aktas H, Braun HJ, Mergoum M, Yildrim AF, Bagci A, Eleckcioglu H, and Yahyaoui, A.

2002. The frequency and diversity of the cyst and lesion nematode on wheat in the Turkish Central Anatolian Plateau.

Nematology 4(2):272.

Nicol JM, Rivoal R, Trethowan RM, van Ginkel M, Mergoum M, and Singh RP. 2001. CIMMYT’s approach to identify

and use resistance to nematodes and soil-borne fungi, in developing superior wheat germplasm. In: Wheat in a

Global Environment (Bedö Z and Láng L eds.). Kluwer Academic Publishers, Netherlands. Pp. 381-389.

Table 2. Germ plasm with field resistance to the root complex after 3 years of field inocula-

tion experiments in Cumra, Turkey. TK = Turkey, TCI = TURKEY/CIMMYT/ICARDA

IWWIP.

Cross Origin

LOV41//LI7/LE2062 Argentina–TCI

Katea-1 Bulgaria–Sadovo

Dachnaya/LAJ3302 TCI

Bilinmiyen96.7 TCI

Burbot-6 TCI

Zargana-2 TCI

Zargana-3 TCI

ECVD12/KAUZ//Unknown TCI

F12.71/SKA//FKG15/3/F483/4/CTK/VEE TCI

F130L1.12/Attila TCI

KRC66/SERI//KINACI79 TCI

KS82W409/SPN//CA8055 TCI

NEMURA/KAUZ//AGRI/NAC TCI

OK81306//ANB/BUC/3/GRK/7C TCI

OK81306/SITTA//AGRI/NAC TCI

Orkinos-1 TCI

Orkinos-3 TCI

PYN//TAM101/AMI/3/KRC66/SERI TCI

Sultan 95 TCI

TAM200/KAUZ TCI

BEZ/TVR/5/CFN/BEZ//SUW92/CI13645/3/NA160/4/EMU/6/UNA TK–Edirne

BEZ/HAWK//ES14 TK–Eskisehir

Cerco/Alondra TK–Eskisehir

ES 14/Flamura 85 TK–Eskisehir

GEREK79 TK–Eskisehir

BLL2973/Thunderbird TK–Konya

DAGDAS TK–Konya

HAWK/AIRI TK–Konya

PLK70/LIRA”S//30-KZ-1 TK–Konya

TX71A1039-VI*3/AMI(TX81V6603)//MVR16-85 TK–Konya

HARA456/4/61-130/414-44//68111/WARD/3/69T02 TK

150

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

ITEMS FROM THE UKRAINE

INSTITUTE OF PLANT PRODUCTION N.A. V.YA. YURJEV

Moskovsky prospekt, 142, 61060, Kharkiv, Ukraine.

The relationship between glume, lemma, and kernel size in polonoid wheats.

R.V. Rozhkov and O.O. Kushchenko.

Polonoid wheats are species that have the traits of T. turgidum subsp. polonicum, including very long (2.5–4.5 cm or

greater) glumes with a straw-like constituency, long lemmas twice the size of the paleas, well-marked knobs on the

rachis under the glume that are lacking in other wheat species, and long large kernels. The polonoid wheat group

includes the naked species T. turgidum subsp. polonicum, T. petropavlovskyi Udacz. et E.Migusch. (2n = 42), and the

hulled wheat T. ispahanicum Heslot (2n = 28). The last species does not have knobs on the rachis. Watanabe (2001)

determined that the gene for the polonoidy traits are on chromosome 7A in T. polonicum and T. petropavlovskyi and 7B

in T. ispahanicum. thus, the polonoidy may have different genetic nature.

Polonoidy may be of practical interest because glumes are one of the photosynthetic organs that are the young-

est and nearest to the kernel, therefore, they may be a significant reserve for providing kernels with nutrients and

determine kernel size. In 2001 and 2002, we studied the polonoid traits displayed in three polonoid species and their F1

hybrids with T. durum cultivar Kharkivs’ka 19 and T. aestivum cultivar Kharkivs’ka 28. The first and second florets of

the medial spikelet were analyzed in at least 25 spikes for each accession or hybrid. We measured the lemma and palea -

length and width of the glume and the length, width, height, and weight of the grain. Paired correlation coefficients (CC)

and degree of dominance (D) were calculated for these traits.

Tables 1 and 2 list the results of the 2-year study. The two tetraploid polonoid species had CCs between

dimensions of glume, lemma and palea and grain length and weight of the second floret that exceeded the index for first

floret and in most cases were nonsignificant. On the other hand, the CCs between glume, lemma, and palea dimensions

for the second floret were less than those for the first. At the same time, almost all the CCs for first floret are greater

than those for the second in the hexaploid polonoid and were moderately to highly significant. In the Kharkivs’ka 19

and Kharkivs’ka 28 cultivars, the CCs were low to moderate and differences between the first and second florets were

not similar for the different trait pairs.

For the F1 hybrid ‘T. ispahanicum/Kharkivs’ka 19’, the CCs for all trait pairs for the second floret of a spikelet

and the most of them for the first floret were high and significant (from 0.71 to 0.94). The CC for one of the most

interesting trait pairs, glume length–grain length, is very high, significant for the second floret (0.94) and moderately

significant for the first (0.58). The CCs for the another interesting trait pair, glume length–grain weight, is high and

significant for the first (0.84) and second (0.74) florets. The CC values indicate heterosis for all trait pairs on the second

floret and for the most of the trait pairs on first floret in ‘T. ispahanicum/T. durum Kharkivs’ka 19’ hybrids in comparison

to the parental lines.

In the ‘T. polonicum/Kharkivs’ka 19’ F1 hybrid, the CC for glume length–grain length in the first floret is

positive, moderate, and significant. For the other trait pairs including grain length and weight, the CCs were not signifi-

cant, moderate, or low. The CCs between glume, lemma, and palea dimensions are high and positive. These parents also

have low, often negative, CCs, whereas the hybrids have positive moderate, though nonsignificant, CCs as a rule.

In the F1 hybrid ‘T. petropavlovskyi/Kharkivs’ka 28’, nearly all CCs including grain length and weight are

nonsignificant for both the first and second florets. The CCs for glume, lemma, and palea are high and significant.

From the three polonoid species, only T. ispahanicum displays dominance by glume length (D = 0.2) in hybrids

with T. durum cultivar Kharkivs’ka 19. In the two other hybrids, ‘T. polonicum/Kaharkivs’ka 19’ and ‘T.

petropavlovskyi/Kharkivs’ka 28’, this trait is inherited as a recessive (D = –0.2 and –0.4 respectively). For grain length

151

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

and weight, dominance of T. polonicum (0.7 and 0.9, respectively), overdominance of T. ispahanicum (1.4 and 2.6

respectively), and recessiveness of T. petropavlovskyi (–0.2 and –0.4, respectively) were observed.

Triticum ispahanicum is dominant over T. durum in lemma length (D = 0.7) and is overdominant for palea

length (3.7). Triticum petropavlovskyi is dominant over T. aestivum for lemma and palea length (the both 1.0) and grain

height (0.3) and is overdominant for grain width (2.0).

Table 1. Correlation coefficients between the morphological traits of spikelets in the polonoid species, T. durum, T. aestivum, and their

F1 hybrids, first floret. Ti = Triticum ispahanicum, Tdp = T. turgidum subsp. polonicum, Tp = T. petropavlovsky, K19 = T. durum subsp.

durum cultivar Kharkivs’ka 19, and K28 = T. aestivum subsp. aestivum cultivar Kharkivs’ka 28. Significant correlations are indicated

with an asterisk (*).

Trait pairs Hybrids and parental forms

Ti F1 Ti/K19 K19 Ttp F1 Ttp/K19 Tp F1 Tp/K28 K28

Glume length–grain length 0.16 0.58 * 0.32 –0.40 0.50 * 0.69 * 0.26 0.21

Glume width–grain length –0.40 0.05 0.08 –0.29 0.38 0.35 –0.02 0.34

Lemma length–grain length 0.35 0.54 0.49 * –0.21 0.42 0.71 * 0.43 * 0.39

Palea length–grain length 0.32 0.32 0.18 –0.18 0.33 0.65 * 0.23 0.47 *

Glume length–grain weight -0.11 0.74 * –0.18 –0.68 * 0.31 0.29 0.20 0.16

Glume width–grain weight 0.13 –0.03 –0.28 –0.72 * 0.30 0.04 –0.26 –0.15

Lemma length–grain weight 0.15 0.68 * –0.01 –0.53 * 0.20 0.39 0.32 0.01

Palea length–grain weight 0.42 0.67 * 0.07 –0.42 –0.11 0.28 0.09 0.49 *

Glume length–glume width 0.48 * 0.36 0.09 0.81 * 0.85 * 0.52 * 0.73 * 0.21

Glume length–lemma length 0.72 * 0.97 * 0.44 * 0.77 * 0.91 * 0.79 * 0.87 * 0.40

Glume length–palea length 0.62 * 0.83 * 0.53 * 0.66 * 0.70 * 0.39 0.71 * 0.27

Lemma length–palea length 0.55 * 0.83 * 0.41 0.60 * 0.71 * 0.55 * 0.78 * 0.06

Table 2. Correlation coefficients between the morphological traits of spikelets in the polonoid species, T. durum, T. aestivum, and their

F1 hybrids, second floret.

Glume length–grain length 0.44 * 0.94 * 0.49 * 0.08 –0.03 0.18 0.42 0.38

Glume width–grain length 0.35 0.71 * 0.15 0.13 –0.01 –0.08 0.21 0.28

Lemma length–grain length 0.63 * 0.84 * –0.03 0.18 0.07 0.37 0.00 0.11

Palea length–grain length 0.39 0.76 * 0.23 0.08 0.18 0.38 –0.12 0.25

Glume length–grain weight 0.04 0.84 * –0.05 –0.20 0.02 –0.28 0.23 0.40

Glume width–grain weight 0.40 0.84 * –0.54 * 0.09 –0.00 0.02 –0.06 0.12

Lemma length–grain weight 0.22 0.85 * –0.20 0.29 0.01 –0.06 –0.05 –0.07

Palea length–grain weight 0.04 0.77 * 0.03 0.17 –0.06 0.05 0.09 0.33

Glume length–glume width 0.41 0.77 * 0.44 * 0.67 * 0.87 * 0.37 0.81 * 0.08

Glume length–lemma length 0.64 * 0.91 * 0.37 0.48 * 0.81 * 0.45 0.78 * –0.12

Glume length–palea length 0.49 * 0.79 * 0.47 * 0.35 –0.17 0.35 0.43 0.17

Lemma length–palea length 0.34 0.92 * 0.58 * 0.44* 0.02 0.71 * 0.64 * 0.57 *

152

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Kharkivs’ka 19 and Kharkivs’ka 28 dominate at various rates for glume width in hybrids with T. ispahanicum

(D = 0.1) and T. petropavlovskyi (0.7). Triticum durum is dominant over T. ispahanicum for grain width (1.0) and height

(0.5), over T. polonicum for lemma length (0.6) and grain width and height (the both 1.0), and is overdominant for glume

width (2.0) and palea length (3.0). Triticum aestivum is dominant over T. petropavlovskyi in grain height (0.3) and

overdominant for grain width (2.0).

Examining glume length with grain length and weight under the growing conditions at Kharkiv, the ‘T.

ispahanicum/Kharkivs’ka 19 durum’ F1 hybrids are early ripening when compared with the durum parent, T. polonicum,

T. aestivum, T. petropavlovskyi, and their hybrids. The grains of T. ispahanicum have time to fully develop, whereas the

other parental lines and hybrids cease grain filling before maturity, which is caused by hot and dry summer temperatures.

A clear relationship exists between the glume dimensions and grain length and weight in hybrids with T. ispahanicum.

Hence, T. ispahanicum seems to by more perspective source of polonoid complex for wheat breeding, than T.

polonicum and T. petropavlovskyi. Moreover, the F1 hybrids of all the three polonoid species are more adaptive than

their parental forms and, as a rule, CCs are more positive and higher in the hybrids than the parents. Improvement may

be gained in grain size (and other related traits) in T. aestivum and T. durum advanced cultivars by means of addition of

polonoid complex if early ripening genotypes are used.

References.

Watanabe N. 2001. Near isogenic lines of durum wheat cultivar LD222 and the origin of T. petropavlovskyi. In: Proc

Internat Conf on Genetic collections, isogenic and alloplasmic lines. Novosibirsk, Russia, 30 July–3 August, 2001.

Novosibirsk Pp. 62-64.

Sowing dates, rates, and phytosanitary state of winter wheat fields.

Yu.G. Krasilovetz, N.V. Kouzmenko, A.E. Litvinov, and V.A. Tzyganko.

These studies were conducted at the Plant Production Institute named after V.Ya. Yuryev (Eastern Forest-Steppe of

Ukraine) in 2001–02. The soil was a typical weakly leached, medium humus, black earth soil. The agrotechnique was

general use. The three sowing dates were 10–13 September, 20–22 September, 30 September–2 October. The sowing

rate was 4.0–5.0 x 106 viable seeds/hectare. The relationship between the phytosanitary state of winter wheat, the sowing

dates, and the sowing rates was studied by preceding black fallow on a manure background, 30 t/ha along with NPK30

application. Plant damage from disease and cereal flies was studied using conventional methods.

In the experimental years, spread and development of root rots (Helminthosporium and Fusarium) and the

intensity of disease development on winter wheat leaves differed for the sowing dates (Table 3). The later sowing data

had a considerable reduction in spread and development of root rots and leaf infection by Septoria in 2001. Leaf rust

occurred less at the first sowing date than at the other two. Damage by powdery mildew was low. The least amount of

shoot damage by cereal flies was observed at the third sowing date; the highest degree was at the second sowing date.

In 2002, the spread and development of root rots did not vary among different sowing dates. The degree of the develop-

ment of Septoria increased with a delay of the sowing date. Leaf rust and powdery mildew were not observed. Damage

to the shoots of winter wheat by cereal flies was not considerable.

Over 2 years, damage by root rots was reduced considerably at later dates of sowing on average. The intensity

of Septoria infection on the upper leaves of winter wheat planted at the first sowing date was 1.6 times lower compared

with the third date. Leaf rust infection in wheat planted at the second and third dates exceeded that in wheat sown at the

first date on all leaves by 2.5–2.6 times and on the upper leaves by 2.2–2.3 times. The lowest degree of shoots damage

by cereal flies was recorded at the third date of sowing winter wheat and highest at the first date.

A shift in sowing date led to changes in the phytosanitary state of winter wheat and, as a result, to the changes in

yield capacity of the crop. Thus, there was a progressive increase in yield from the first to the third sowing dates in 2001

and, a decreased grain yield with a delay of planting in 2002. On average over 2 years, grain yield increased by 2.6–7.6

c/ha at the third sowing date in comparison with that at the second and first sowing dates.

153

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

The effect of sowing rates on the phytosanitary state of winter wheat planting is given in Table 4. In 2001, we

noted that the sowing rate of 4–5 x 106 viable seeds/ha did not differ with respect to the damage by diseases and cereal

flies. The 2002 data showed that an increase in sowing rates from 4–5 x 106 viable seeds increased damage by root rots

and Septoria. Powdery mildew and leaf rust were not factors in 2002.

The spread and development of root rots at the increased sowing rate did not increase considerably on average

over 2 years. Septoria infection on all leaves of the winter wheat plants in these variants was approximately the same as

that of the upper leaves for planting rates with 5 x 106 seeds; exceeding this index compared with a usual rate of 4 x 106

Table 3. Phytosanitary state of winter wheat plants depending on sowing dates.

Root rots at Powdery

tillering (%) Septoria mildew Leaf rust Shoot

damage Grain

Sowing develop- all upper all upper all upper by cereal flies yield

date spread ment layers layer layers layer layers layer (%) (c/ha)

2001

I 34.6 13.2 36.6 7.5 0.3 0.2 5.1 6.6 3.0 48.9

II 23.1 8.6 26.0 3.4 1.1 0.4 13.1 15.1 3.7 60.9

III 12.8 4.6 24.0 3.4 1.4 0.1 13.6 14.5 0.7 72.2

LSD05 — 5.3 — 2.6 0.7 — 7.1 7.2 0.8 —

2002

I 48.7 21.3 46.0 18.2 0.0 0.0 0.0 0.0 2.0 78.6

II 49.7 24.5 52.0 30.6 0.0 0.0 0.0 0.0 0.6 76.7

III 47.0 25.1 53.5 36.7 0.0 0.0 0.0 0.0 0.0 70.6

LSD05 — 6.8 7.8 14.9 ———— 0.8 —

Mean (2001–02)

I 41.7 17.3 41.3 12.9 0.2 0.1 2.6 3.3 2.5 63.8

II 36.4 16.6 39.0 17.0 0.6 0.2 6.6 7.6 2.2 68.8

III 29.9 14.9 38.8 20.1 0.7 0.05 6.8 7.3 0.4 71.4

Table 4. Phytosanitary state of winter wheat on black fallow depending on a sowing rate (N30P30K30).

Root rots at Powdery

tillering (%) Septoria mildew Leaf rust Shoot No. of

damage productiveGrain

Sowing develop- all upper all upper all upper by cereal flies tillers yield

date spread ment layers layer layers layer layers layer (%) (m2)(c/ha)

2001.

4.0 34.6 13.2 36.0 7.5 0.2 0.1 3.5 5.5 2.5 601 47.4

5.0 34.9 13.5 36.6 7.5 0.3 0.2 5.1 6.6 3.0 701 48.9

LSD05 —— ——0.1——— — ——

2002.

4.0 48.7 21.3 46.0 18.2 0.0 0.0 0.0 0.0 2.0 664 78.6

5.0 67.6 27.4 53.4 31.8 0.0 0.0 0.0 0.0 1.2 690 84.6

LSD05 —— 3.43.0———— — ——

Mean (2001–02).

4.0 41.7 17.3 41.0 12.9 0.1 0.05 1.8 2.8 2.3 633 63.0

5.0 51.3 20.5 45.0 19.7 0.2 0.1 2.6 3.3 2.1 696 66.8

154

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

seeds. The variants with different sowing rates did not vary greatly in relation to the damage of shoots by cereal flies. In

winter wheat sowings at 5 x 106 viable seeds/ha, the number of productive tillers increased and, as a result, grain yield

was higher by 3.8 c/ha compared to the yield at 4 x 106 seeds/ha.

Productivity of spring durum wheats at different seeding rates.

O.V. Golik.

Creating new spring durum wheat cultivars demands a variety of agrotechnics for obtaining the highest yield while

maintaining high grain quality. Spring wheat in the Ukraine is grown in risky agricultural conditions. Moisture is the

main limiting factor and an optimal seeding rate is the main factor for greatest yield. According to most authors, crops

with optimal and dense seeding rates ensure the largest grain yield in any agroecological conditions (Makrushin 1985;

Sechnyak et al. 1983). Overgrown plants in thin crop stands have an increased vegetative period and a higher degree of

infection by fungal diseases and pests. Thinning crops promotes weed growth. All of these factors decrease the seed-

sowing quality (Chulkina et al. 2000). Lelli (1980) stated that the productivity potential of a plant is hereditary and

depends on genetically and ecological conditions. Studying the elements of yield structure is a prerequisite of this

potential. Thus, we wanted to investigate and determine optimal seeding rates for new cultivars bred for the conditions

of the eastern Forest-Steppe Region of the Ukraine.

We analyzed the spring durum wheats Kharkovskaya 15 and Kharkovskaya 23 (standards of the Ukrainian state

Service on right protection for plant cultivars), Kharkovskaya 46 (grain quality standard), and Kharkovskaya 19 (lodg-

ing-resistance standard) and the new cultivars Kharkovskaya 27, Kharkovskaya 33, and Kharkovskaya 41 under the

climatic conditions of the eastern Ukraine (Kharkov) between 1998 and 2000. The characters analyzed included

productivity (g/m2), grain yield (%, the ratio weight of kernels to weight of chaff), plant stand (plant/m2), productive

tillering, and 1,000-kernel weight (g) with seeding rates of 3, 4, 5, 6, and 7 x 106 germinating kernels/hectare (MKH).

The humidity varied over the years; 1998 and 1999 were severe droughts and 2000 was optimal but irregular in different

vegetative phases. The 1-m2 plots were replicated three times. The cultivars were sown in an experimental field

following peas. The results are from a three-factor dispersion analysis.

No differ-

ences were observed

for durum wheat with

different seeding rates.

The productivity

exceeded the general

mean (155 g/m2) at

seeding rates of 3, 5,

and 7 MKH (172–175

g/m2) only in 2000.

The least productivity

was 146 g/m2 in 1998.

The highest mean

productivity (170 g/

m2) was found in

Kharkovskaya 33 at 3

MKH (Table 5

represents data for

only for best

(Kharkovskaya 33)

and worst

(Kharkovskaya 19)

cultivars. The

productivity of these

cultivars was equal to

or higher than the

general mean in all

Table 5. The influence of seeding rates on productivity and associated traits in durum wheat

cultivars, 1998–2000. * = reliable in comparison with the means by cultivars, ** = reliable

in comparison with means by experiment (by trait), and *** = reliable means by cultivar.

The mean indices of traits

Seeding Produc- Grain Plant 1,000-

rate tivity yield stand Productive kernel

Cultivar (x 106/ha) (g/m2)(%) plants/m2tillering weight (g)

Kharkovskaya 33 3 183 * 31.4 172 * 0.99 34.5 *

4 157 32.2 197 * 1.00 * 34.6 *

5 184 * 30.6 236 1.00 * 33.5

6 150 29.8 259 * 0.93 31.4 *

7 179 * 31.8 292 * 1.01 * 31.0 *

Mean by cultivar 170 ** 31.2 231 0.99 33.0 **

Kharkovskaya 19 3 113 * 29.6 146 * 0.91 * 32.7 *

4 154 29.7 173 * 0.92 * 33.6

5 135 27.1 * 209 * 0.91 * 32.2 *

6 128 * 29.2 234 0.89 * 31.2 *

7 130 * 28.9 264 * 0.89 * 32.1 *

Mean by cultivar 132 ** 28.9 ** 205 ** 0.90 ** 32.4 **

Mean by experiment 155 30.8 227 0.96 33.9

Least significant difference 5 % 9.4 1.06 11.9 0.037 0.53

155

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

variants. The highest value was 233 g/m2 at 7 MKH for Kharkovskaya 33 in 2000. Thus, Kharkovskaya 33 was the

most productive cultivar grown in the climatic conditions of the Ukraine. The basic group of tested wheats

(Kharkovskaya 15, Kharkovskaya 23, Kharkovskaya 27, Kharkovskaya 41, and Kharkovskaya 46) had a productivity

equal to the general mean. The optimal seeding rate was 4 MKH. The productivity of Kharkovskaya 19 was minimal

(100 g/m2 in 1999 at 3 MKH). The optimal seeding rate was 5 MKH.

Grain yield depended more on climactic condition. Few cultivars differentiated and did not depend on seeding

rates. Those exceeding the general mean of 30.8 % for all cultivars in 1998 (35.7 %) were less in 2000 (25.3 %). The

grain yield of Kharkovskaya 23 (33.2 %) and Kharkovskaya 27 (33.5%) reliably exceeded the general mean of

Kharkovskaya 19 (28.9 %) and Kharkovskaya 46 (27.8 %) usually less at all seeding rates. This character of other

varieties was up-to-date of general mean.

The 1,000-kernel weight also depended more on climatic conditions and less on cultivar differences and seeding

rates. This trait always exceeded (34.4–36.5 g) the general mean (33.9 g) in 1998 and 1999 but was lower (30.7 g) in

2000. Kharkovskaya 23 and Kharkovskaya 27 reliably exceeded (35.2–35.3 g) the general mean for all cultivars. Little

difference was observed in the 1,000-kernel weight at different seeding rates.

Variability in productive tillering was similar to 1,000-kernel weight but reliable only in some years (1.03 in

1999, 0.89 in 2000, 0.9 for Kharkovskaya 19, and 1.02 for 3 MKH, compared to the general mean of 0.96. Plant stand

depended on seeding rate (rates of 3 and 4 MKH reliably exceeded the general mean of 227 plant/m2) and equaled 160–

198 plant/m2; 5, 6, and 7, MKH were 237–283 plant/m2. Less reliable were years and cultivar.

Graphics can simplify the dependence estimates of

the tested characters by specific factors. For example,

productivity in specific ecological conditions by different

seeding rates can be represented with the help of a quadratic

surface. If the ecological conditions are a mean value of

productivity during a suitable year, the maximum of the

surface indicates the highest possible display this character

(see Figs. 1 and 2).

Thus, the effect of climatic conditions on traits was

greatest on 1,000-kernel weight, productive tillering, and

grain yield for the variables productivity and plant stand.

The lack of difference in productivity and grain yield and the

minimal difference in 1,000-kernel weight and productive

tillering at different seeding rates indicates the impossibility

for estimating the optimal seeding rate for durum wheat.

Therefore, this question can be solved only for the concrete

cultivar. Kharkovskaya 33 was the most productive with a

mean index of other testing traits. The optimal seeding rate

was 5 MKH. Kharkovskaya 23 and Kharkovskaya 27 had

higher grain yield and 1,000-kernel weight with mean indices

of other testing traits. The optimal seeding rate was 4 MKH.

Kharkovskaya 41 has mean indices for all traits. The optimal

seeding rate was 5 MKH. This cultivar was entered in the

Kharkovskaya 46 have most indices of plant stand, productive tillering, and mean other traits. The optimal seeding rate

was 5 MKH. Kharkovskaya 19 had the smallest indices for all tested traits. The optimal seeding rate was 4 MKH.

References.

Chulkina VA, Toropova EYu, Chulkin YuI, and Stezov GYa. 2000. The agrotechtical method of plant protection.

Novosibirsk:UKEA Publishing, 241 pp. (in Russian).

Lelli Ya. 1980. The breeding of wheat. Moscow:Kolos, 383 pp. (in Russian).

Makrushin NM. 1985. Ecological basis of industrial seed-growing of cereals. Moscow: Agro Industrial Publishing, 139

pp. (in Russian).

Sechnyak LK, Kindruk NA, Slusarenko et al. 1983. The ecology of wheat seeds . Moscow:Kolos, 164 pp. (in Russian).

Fig. 1. Productivity

of Kharkovskaya 19

for seeding rates and

climatic conditions,

1998–2000. Mean

productivity of the

cultivar in suitable

years.

Fig. 2. Productivity of

Kharkovskaya 33 for

different seeding rates

and climatic

conditions, 1998–

2000. Mean

productivity of the

cultivar in suitable

years.

156

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Winter wheat gene pool of the CIMMYT international nurseries for improvement of breeding for

resistance to fungal diseases and productivity in the eastern Forest-Steppe Region of the Ukraine.

V.V. Sotnikov and T.M. Yevlanova.

We conducted a soil-quarantine control on quarantine pathogens and a primary investigation of 164 bread and 30

samples of hard winter wheat from the CIMMYT International Nurseries at the Introductory Quarantine Nursery of the

Plant Production Institute in 2001–02. We looked at the main economical and morphological characters, biological

properties, and resistance to nonquarantine, harmful organisms in order to replenish the stocks of the National Centre for

Plant Genetic Resources of Ukraine and breeding subdivisions of the Institute with a new foreign germ plasm that is

fully free of quarantine pathogens and for the use of highly productive cultivars in breeding. The introductory material

was from the 5th Winter Wheat Observation Nursery for Semi-Arid Areas (5th WON-SA, 102 entries), 6th Facultative

and Winter Wheat Elite Yield Trial for Rainfed Areas (6th EYTRF, 20 entries), 4th Winter Wheat East-European Re-

gional Yield Trial (4th WWEERYT, 42 entries), and 3rd Winter Durum Wheat East-European Regional Yield Trial (3rd

WDEERYT, 30 entries), which contained cultivars from Azerbaijan, Bulgaria, the Czech Republic, Georgia, Hungary,

Iran, Kazakstan, Moldova, Romania, the Russian Federation, Syria, Turkey, the Ukraine, and the U.S.A.

The bread wheat cultivar Albatros odeskiy and the hard wheat Kharkivska 32 were used as local checks. The

degree of damage was estimated after winter according to a 0–9 scale: 0 = death, 1 = very low, and 9 = very high. The

same scale was used to estimate resistance to fungal pathogens in natural conditions where 1 = very high susceptibility, 5

= moderate susceptibility, 6 = moderate resistance, and 9 = very high resistance. Yield was considered very low if it was

less than 76 % of the local checks Albatros odeskiy and Kharkivska 32, low = 76–95 %, average = 96–115 %, high =

116–135 %, and very high > 135 %. Thousand-kernel weight was low if less than 39.0 g, average = 39.0–46.9, high =

47.0–54.0, and very high > 54.0 g. Planting in the 5th WON-SA and 6th EYTRF was done with a hand-sower at the

planting rate of 60 seeds/m in 0.75-m2 plots. Planting of the 4th WWEERYT and 3rd WDEERYT was done with a

tractor-sower SSFK-7 at a planting rate of 6.0 x 106 viable seeds/ha in 8-m2 plots. Vitavax was used as a seed treatment.

Field trials were conducted in arid conditions with a black fallow forecrop. The planting date was very late, 5 October,

2001, or 17 days later than permissible (18.09). Overwintering was satisfactory because of a mild winter. Growth

recovery in the spring began much earlier than in comparison with the mean sowing dates of earlier years. Yield level

was limited by the late sowing date; an outbreak of fungal diseases including powdery mildew, leaf rust (only on the

bread wheats), and Septoria blight; and unfavorable climactic factors including increased daily temperature, hot dry

winds, and the lack of soil moisture from the end of tillering to grain filling. Over that period, rainfall was 84 mm (66 %

of the mean of many years). During a entire vegetative period, rainfall was approximately 260 mm (105 % of the yearly

average). Total rainfall, including winter precipitation, was 408 mm. In spite of these negative effects, the wheats

produced a high grain yield. The average yield capacity of the local checks and control cultivars in the 5th WON-SA and

in the 6th EYTRF nurseries were Albatros odeskiy, 414 g/m2; Donetska 48, 534 g/m2; Kharus, 466 g/m2; Odeska 267,

485 g/m2; Kharkivska 96, 627 g/m2; Tira, 465 g/m2; Gerek 79, 370 g/m2; Dagdas 94, 438 g/m2; Suzen 97, 408 g/m2;

Kirgiz 95, 541 g/m2; and Gun, 244 g/m2.. In the 4th WWEERYT, average yield capacities were Albatros odeskiy, 277 g/

m2; Donetska 48, 335 g/m2; Kharus, 415 g/m2; Myronivska 61, 384 g/m2; Bezostaya 1, 280 g/m2; Seri, 199 g/m2; Jagger,

246 g/m2; and in the 3rd WDEERYT Kharkivska 32, 314 g/m2; Aisberg odeskiy, 284 g/m2; and Kunduru, 177 g/m2.

A mild winter did not permit evaluation of the entries for winter hardiness. A predominant part of the material

had a high degree of wintering (scores of 8–9), with the exception of ‘494J6.11//TRAP#1/BOW’ (wintering score = 4);

Unknown 95-3 (3); ‘FRTL//AGRI/NAC (2)’; ‘Nemura/Kauz//AGRI/NAC (TOP SIEVE95-TOP SIEVE96 TOP

SIEVE97)’ (5.5); all entries from Turkey; the 5th WON-SA nursery; ‘Saulesku #44/TR810200’ (5) from Turkey; the 6th

EYTRF; ‘Brindur/DF 38-86’ (5); ‘DF900-83/WPB881’ (U.S.A) and Altin (4.5); DUT-TA00-22 (1); ‘DICLE74/

HALKALI058’ (5) from Turkey; DYT-CA00-7 (5) from Syria; Turan (1) from Azerbaijan; and the 3rd WDEERYT

nursery.

The heading of the bread wheats for the 5th WON-SA compared to the local check Albatros odeskiy (headed

144 days after 1 January) were 55 % similar and 38 % earlier and 41 % and 59 %, respectively, yielded greater than the

check. In the 6th EYTRF, 70 % were earlier and 30 % were similar and 86 % and 67 %, respectively, yielded greater

than the check. In the 4th WWEERYT, 67 % were similar and 31 % were earlier; the number of entries yielding greater

than the check were 82 % and 69 %, respectively. For the hard wheats, when compared to the check Kharkivska 32 (147

days from 1 January) in the 3rd WDEERYT 43 % were similar, 30 % later, and 27 % earlier and those entries with a

yield of 96 % of the check were 8 %, 11 %, and 50 %, respectively.

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

Leaf fungal diseases were caused considerable damage. Susceptibility (1–4 scale) was estimated in the nurser-

ies (Table 6). This natural infection permitted differentiation of the material for the degree of resistance to the major

pathogens. A large part of the entries in the 6th EYTRF nursery (83 %) had average or higher grain yields than the

check. In the 4th WWEERYT, yield was 76 %, 45 % in the 5th WON-SA, and 21 % in the 3rd WDEERYT compared to

the check.

We distinguished the following entries that had a combination of useful economic characters and resistance or

moderate susceptibility to leaf disease pathogens. These lines are recommended for further study and possible use in

breeding highly productive cultivars of bread and hard wheats. Unfortunately, most lines of these group have resistance

to two disease pathogens but considerable susceptibility to a third, making their use difficult. The combination of

economical characters given include 1 = mean yield, 2 = high yield, 3 = very high yield, 4 = average 1,000-kernel

weight, 5 = high 1,000-kernel weight, 6 = very high 1,000-kernel weight, 7 = heading ≥ 2 days earlier compared to the

check, and 8 = similar heading date as check.

Moderately susceptible or resistant to powdery mildew and Septoria but highly resistant to leaf rust. CIT90089-

0YC-0YC-0YC-7YC-0YC-1SE-0YC-4YC-0YC (4, 7; pedigree: Weston/VEE) and BDKE930161-0YC-0YC-1YC-

0YC-3YC-0YC (2, 4, 7; pedigree: Haymana75/4/YMH/TOB//MCD/3/LIRA(BDME 9)). Both these lines were from

Turkey and the 5th WON-SA nursery.

Moderately susceptible to powdery mildew and Septoria. MVTD 15-99 (4; Hungary, 3rd WDEERYT nursery,),

CIT932314-0SE-0YC-2YE-0YC-2YK-0YK (1, 4, 8; pedigree: RSK/FKG15//CHAM6/3/FDL4; Turkey, 5th WON-SA,),

and CIT94072-0SE-1YC-0YC (3, 4, 8; pedigree: PYN//TAM101/AMI/3/KRC66/SERI; Turkey, 6th EYTRF).

Moderately susceptible or resistant to powdery mildew and leaf rust. SG-RU 8069 (3, 4, 8; Czechia), Iveta NTA-92/

89-6 (2, 7; Bulgaria); GK Bagoly (1, 4, 8; Hungary), GK Vevecky (2, 4, 8; Hungary), MV Dalma (2, 4, 8; Hungary), GK

Forras (3, 8; Hungary), MV 04-96 (3, 5, 8; Hungary), Turda 2000 (2, 4, 8; Romania), Destin (2, 4, 8; Romania), Efekt (1,

4, 8; Romania), Expres (2, 4, 7; Romania), Manypa (3, 8; Moldova), Strumok (3, 4, 8; Ukraine), Erythrospermum 270 (3,

7; Ukraine), Knjazhna (3, 4; Russia), and Akinci-84 (3, 4, 8; Azerbaijan). All entries were from the 4th WWEERYT

nursery.

Moderately susceptible to Septoria and resistant or moderately susceptible to leaf rust. CIT922411-0SE-0YC-

2YC-0YC-2YC-0YC-3YC-0YC (1, 4, 7; pedigree: CHAM4/TAM200//RSK/FKG15), CIT90089-0YC-0YC-0YC-7YC-

0YC-1SE-0YC-3YC-0YC (1, 4, 7; pedigree: Weston/VEE), CIT937256-0SE-0YC-3YE-0YC-1YC-0YC (2, 4, 8;

pedigree: PLK70/LIRA//Attila/3/AGRI/NAC), CMSW93WM0071-0AP-0YC-8YE-0YC-3YC-0YC (1, 4, 8; pedigree:

FRTL//AGRI/NAC), CIT922229-0SE-0YC-1YC-0YC-7YC-0YC-2YC-0YC (2, 4, 8; pedigree: Necomp1/5/BEZ//TOB/

8156/4/ON/3/TH*6/KF//), and MXTK930076-0SE-0YC-12YE-0YC-4YK-0YK (1, 5, 7; pedigree: 1D13.1/MLT/3/LFN/

SDY//PVN). All entries are from Turkey and the 5th WON-SA nursery. CIT945175-030SE-0YC-7YE-0YC (3, 4, 7;

pedigree: DDZ2141.85.271/ES14//F134.71/NAC) from Turkey and the 6th EYTRF nursery.

Resistant to powdery mildew. ELIDUR (1, 4, 7; Romania) and Perlyna (2, 5, 8; Ukraine) both from the 3rd

WDEERYT nursery; CIT935011-0SE-0YC-3YE-0YC-2YC-0YC (1, 4, 7; pedigree: ES14/130L1.12//MNCH) and

CIT930082-0SE-0YC-3YE-0YC-2YK-0YK (3, 5, 8; pedigree: KARL/Ariesan) both from Turkey and the 5th WON-SA

nursery; ICWH900747-0AP-0YC-0YC-6YC-0YC-9YC-0YC (1, 4, 7; pedigree: Motah-7, Turkey, 6th EYTRF); Demir

(2, 5, 7, Turkey, 4th WWEERYT); Kiziltan (5, 8; Turkey, 3rd WDEERYT); and Ankara 98 (6, 8; Turkey), Yilmaz (5, 8;

Table 6. Overall susceptibility scores of each of the international nurseries investigated in 2001–02.

Powdery mildew Septoria Leaf rust

5th Winter Wheat Observation Nursery for Semi-Arid Regions 70 62 28

6th Elite Yield Trial for Rainfed Regions 90 80 45

4th Winter Wheat East European Regional Yield Trial 36 98 12

3rd Winter Durum East European Regional Yield Trial 17 97 no damage

158

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Turkey), Brindur/DF 38-86 (1, 4, 7; accession #019006; U.S.A.), Brindur/DF 38-86 (2, 4, 7; accession #019007;

U.S.A.), UVY162/61.130//HC6654/3/AKB/OVI65/4/WPB881 (5, 8; U.S.A.), WPB881/Rodur (1, 4, 8; U.S.A.), and

WPB88/H7092-50B//MI83.84.503 (5; U.S.A.) all entries from the 3rd WDEERYT nursery.

Moderately susceptible to Septoria. CIT922142-0SE-0YC-3YC-0YC-6YC-0YC-1YC-0YC (2, 4, 7; pedigree: JI5418/

MARAS), CIT88088T-0SE-1YC-0YC-2YC-0YC-2YC-0YC-8YC-0YC-1YC-0YC (2, 5, 8; pedigree: Zander-34), and

CMSW93WM0182-0AP-0YC-5YE-0YC-1YC-0YC (1, 4, 8; pedigree: SW89-3218//ASP/BLT) all from Turkey and the

5th WON-SA nursery; and CMSW94WM00586S-03Y-0B-0SE-1YE-0YC (2, 5, 7; pedigree: Saulesku #44/TR810200)

from Turkey and the 6th EYTRF nursery).

Resistant to leaf rust. Capuz (2, 4, 8; Moldova), Kupava (3, 4, 8; Russian Federation), and Kroshka (3, 4, 7; Russian

Federation) all from the 4th WWEERYT nursery; 0YA-0YA-5YC-0YC-6YC-0YC (1, 5, 8; pedigree: BUC/5/Naphal/

CI13449/4/SEL14.53/3/Lancer//ATL66/CMN), CIT922142-0SE-0YC-3YC-0YC-6YC-0YC-2YC-0YC (3, 4, 7; pedi-

gree: JI5418/Maras), CIT932332-0SE-0YC-7YE-0YC-3YK-0YK (2, 5, 7; pedigree: CHAM6//1D13.1/MLT/3/

SHI4414/CROW/4/KVZ/AU//GRK), CIT932282-0SE-0YC-3YE-0YC-3YK-0YK (1, 5, 8; pedigree: Karous-10),

CIT935166-0SE-0YC-4YE-0YC-1YK-0YK (3, 4, 7; pedigree: PLK70/LIRA/5/NAI60/3/14-53/ODIN//CI13441/4/

GRK/6/MNCH), 0YA-0YA-5YC-0YC-4YC-0YC (3, 5, 7; pedigree: BUC/5/NAPHAL/CI13449/4/SEL14.53/3/L//

ATL66/CMN), and CIT935224-0SE-0YC-3YE-0YC-3YC-0YC (3, 5, 8; pedigree: NGDA146/4/YMH/TOB//MCD/3/

LIRA/5/F130L1.12) all from Turkey and the 5th WON-SA nursery; F2.96.24-0SE-0YC-1YE (2, 4, 8; pedigree:

Bilinmiyen96.24), CIT932332-0SE-0YC-1YE-0YC-2YC-0YC (3, 4, 7; pedigree: CHAM6//1D13.1/MLT/3/SHI4414/

CROW/4/KVZ/AU//GRK), and CIT930151-0SE-0YC-9YE-0YC-1YC-0YC (3, 5, 7; pedigree: Jing Dong 1//1D13.1/

MLT) all from Turkey and the 6th EYTRF nursery.

Immunological basis for developing initial material resistant to septoriosis for winter and spring

bread wheat breeding in Ukraine.

S.V. Rabinovich, V.P. Petrenkova, I .M. Chernyaeva, and L.N. Chernobay.

Among the cereal crops, wheat is the major grain grown in Ukraine. The winter wheat area occupies about 7 x 106

hectares; spring wheats occupy 3–3.5 x 105 ha. Resistance to disease pathogens for increasing the yield capacity of

cereal crops in developed countries has become more significant than other traits. The acuteness of the problem will not

decrease in the future as breeding progress for productivity effects a pathogen’s development and accelerates adaptation.

The breeder, however, must take the lead in order to ensure constant sources and donors of disease-resistance genes.

Septoria resistance will play an important role among other diseases. The best results will be achieved only by the use of

resistant material in breeding.

Septoria has been one of the most harmful diseases of wheat during the last decade, although it has been known

since 1907 (Yachevskiy 1908). At present, this disease is widespread all over the regions where wheat is cultivated,

including the Ukraine. The disease symptoms appear as spots, occurring on all above-ground parts of the plant and at all

developmental stages. The disease pathogens in the Eastern Forest-Steppe of Ukraine include S. tritici and S. graminum,

which affect the leaves and leaf sheaths of winter and spring wheats (Boublik et al. 1999).

The ascospores are an additional source of infection, but the pycnospores are of great importance in infecting

and reinfecting plants. Prolonged moisture, mild, windy weather, and precipitation, especially at the heading and

flowering, are favorable to infection. At the Department for the Plant Immunity to Diseases and Pests in the Plant

Production Institute named after V. Ya. Yurjev between 1996–2002, we studied some genotypes from a collection of

breeding material of winter and spring wheats obtained from the National Centre for Plant Genetic Resources of Ukraine

(NC PGRU) and breeding departments of the Institute. Artificial infections for Septoria infection were created according

to known procedures (Anonymous 1989a and b). In the Ukraine, a parasite of the Septoria pathogens at the ascospore

stage has been found in cereals that belongs to the genus Leptosphaeria Ces. et de Not.

The years of the study were characterized by considerable changes in weather conditions, particularly during

plant development; 1997 and 2000 were favorable for plant and pathogen development characterized by increased

moisture and mild temperatures, and 1996, 1998, and 1999 were years of severe drought during the entire vegetative

period of both winter and spring cereals and a reduced level of pathogen development. The weather conditions in 2001

159

A n n u a l W h e a t N e w s l e t t e r V o l. 4 9.

were characterized by increased moisture

at the start of plant vegetation and

flowering stages and severe drought and

heat during grain formation and filling.

Epidemics of the disease in 1997 and

2000 did not require creating artificial

infections.

The maximum infection of

plants amounted to 65–100 %. In 1996,

the pathogen was epidemic among the

leaf diseases. In 1998, Septoria in winter wheat was suppressed at the milk stage. In 1999, fungal pycnidia emerged in

the first week of May on the bottom leaves of plants, spread to the middle and upper leaves, but symptoms were not

found on the flag leaf due to a severe drought (Table 7). In 2001, favorable conditions for disease development in both

winter and spring wheats were produced by artificial infections through use of a local population and a population

received from the Plant Protection Institute (Kyiv). During 1996–2000, we studied the resistance of 1,542 samples of

winter wheat and 1,186 samples of spring wheat. In 2001 with artificial infection, 453 lines of winter wheat and 202

lines of spring wheat were tested.

Table 8. Resistance to Septoria in a collection of winter bread wheats at the Department for Plant Immunity to

Diseases and Pests in the Plant Production Institute named after V.Ya. Yurjev between 1996–2001. Scores are on a

scale of 1–9 where 0 = susceptible, 5 = medium resistance, and 9 = high resistance; — indicates no test.

Resistance score by year

Cultivar Origin 1996 1997 1998 1999 2000 2001

Myronivska 32 Ukraine 5 5 7–6 7 — —

Myronivska 33 Ukraine 9 8 8 7 6 6

Myronivska 64 Ukraine 7 5 7–6 7 — —

Myrich Ukraine 7 — 7–6 7 6 5

Kyivska 7 Ukraine 6 — 8 6 6 5

Luna 3 Ukraine 8 6 6 6 7 —

Lutescence 20191 Ukraine — — 7–6 6 7 6

D 169 Ukraine — — 7 7–6 6 5

Atol Odeskiy (T. durum)Ukraine — — — 8 8 6

Plamya Ukraine 6 — 7–6 7–6 5 5

Don 93 Russia — — 6–5 6 5 5

Smouglyanka Russia — — 6 6 6 6

Knyazhna Russia — — 7–6 7 5 5

Arbatka Russia — — 7 6 6 5

Norman Great Britain — — — 7 8 8–7

Arina Czech Republic 8 6 6 — — —

Ikarus Austria 7 6–7 7 — — —

MV23 Hungary — 6 7–6 6 — —

Granada Germany 8 6 7 — — —

Niclas Germany 8 7 7–8 — — —

Olma Poland — — — 6–7 7 6–7

SMH 2893 Poland — 6 7–6 7 6 —

Panda Poland 7 6–7 6–7 — — —

N92L228 U.S.A — — 7–6 6 6 5

Wakefield U.S.A — — — 6–7 6 5–7

KS91WGRC11 U.S.A — 7 — — 7 6

Charmany U.S.A — 6 7–6 — 7 6

TX90V8727 U.S.A — 6 — 8 — —

Table 7. Maximum infection of cereal crops by Septoria, 1996–2001.

Year of study

Crop 1996 1997 1998 1999 2000 2001

Winter wheat 65.0 100.0 60.0 25.0 100.0 65.0

Spring bread wheat 21.0 100.0 65.0 14.0 100.0 65.0

Spring durum wheat 25.0 100.0 25.0 20.0 65.0 40.0

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A n n u a l W h e a t N e w s l e t t e r V o l. 4 9 .

Immune or highly resistant lines were not identified, which shows the low adaptation of resistance genes in the

cultivars studied. In the material of the competitive variety trials of the Winter Wheat Breeding Department, two lines

were the most resistant (scores 6–7), Lutescens 159-95 and Erythrospermum 224, and 10 lines were of medium resis-

tance (score 5). The rest of the material from the nursery was susceptible or very susceptible (Chernyaeva and Mouraeva

1992; Dolgova et al. 1997; Rabinovich et al. 1999).

Among samples from the world gene pool of winter wheat, we identified genotypes that maintained resistance

to the pathogen (scores 6–8) including Granada from Germany and Myronivska 33 and Myrich from the Ukraine.

Myrych and Myronivska 33 of Myronivskiy Institute of Wheat n.a. V.M. Remeslo were used as initial material for

developing winter wheat resistant hybrids (see Table 8).

Septoria infection in spring wheat was at a lower degree than that of winter wheat. Spring wheats also were

infected by leaf rust and powdery mildew fungi. We identified medium-resistant breeding lines (scores 5–8) between

1990–96. In 2002, line 97-171 showed medium resistance.

Among the durum wheats, we identified three lines that maintained resistance during 3 years at a level of 6–7;

Leucurum 79, from Kazakhstan, and Hordeifome 1613 and Hordeiforme 1620 from Bulgaria. In 2000, no resistant lines

were identified and only two cultivars, Irridur (U.S.A.) and CD 89239 (Mexico), were medium resistant (score 6).

Both spring bread and winter wheats had high levels of Septoria infection. No immune or resistant lines were

found after many years. Five genotypes were classified to be of medium resistance (score 5; Largo and Oasis (U.S.A.),

and Krasnokutskaya 9, Legenda, and Lutescens 115/85-3 (Russian Federation). In the 2000 epidemic, three lines from

the Samariskiy NIISHK scored 6 (medium resistant); Volgouralskaya, Erythrospermum 1508, and Erythrospermum

1509.

Disease dynamics were investigated to analyze resistance in breeding material in an artificial infection. Resis-

tance was determined by the AUDPC. According to the criteria, the best lines were the winter wheats Lutescence 234-

99, Erythrospermum 293-99, Lutescence 422-2000, and Lutescence 625-2000 with resistance scores of 6. Infection did

not exceed 15 %, with the average indices of resistance at scores 4–5.

Among the NC–PGRU genotypes useful as the sources of resistance were the Ukrainian winter wheats AC-182,

Lutescence 20191, TK 121 Line 2, Perlyna Lisostepu, Myronivska 67, and Myronivska 68; cultivars from Great Britain

Tara, Brigadier, Hussar, and Norman; the U.S. wheats Wakefield, U 1254, KS91WGRC11, and Charmany; the Russian

cultivars Smouglyanka, Douslyk, and Delta; and the Polish cultivar Olma.

References.

Anonymous. 1989a. Methods for estimation of resistance of breeding material and wheat varieties to septoriosis.

Moscow 43 pp.

Anonymous. 1989b. Methods for breeding and estimation of resistance of wheat and barley to diseases in the CEA

member-countries. Prague 321 pp.

Boublik LI, Vasechko GI, Vasyljev VP, et al. 1999. Plant Protection Reference Book (Losovoi MP and Urozhai K eds).

744 pp.

Chernyaeva IM and Mouraeva OV. 1992. Search for sources of resistance for breeding Septoria-resistant varieties of

winter wheat. In: Heads Rept Internat Conf, Scientific bases for stabilization of the production of plant breeding

produce. Jalta (in Ukrainian).

Dolgova OM, Chernyaeva IM, Man’ko OP, and Afons’ka OYu. 1997. Resistance of winter wheat in the Eastern Forest

– Steppes of the Ukraine to Septoria tritici. Ann Wheat Newslet 43:240-241.

Rabinovich SV, Afons’ka OYu, Dolgova OM, and Chernyaeva IN. 1999. Development of wheat cultivars with genetic

protection against diseases–the basis for grain stable production. In: Heads Rept Internat Conf, Scientific bases for

stabilization of the production of plant breeding produce. Kharkiv (in Ukrainian).

Yachevskiy. 1908. Annual news on diseases and damage of cultivated and wild useful plants. S. Petersbourgh. 2,006

pp.

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