GrainGenes

ITEMS FROM HUNGARY

AGRICULTURAL RESEARCH INSTITUTE OF THE HUNGARIAN ACADEMY OF SCIENCES

Martonvásár, H-2462, Brunsvik str. 2, Hungary.

www.mgki.hu

The wheat season.The weather for the 2005-06 crop season was basically favorable for winter wheat. The winter was milder than usual at first, but January 2006 temperatures were below average. However, the snow cover prevented any great damage to the crops. Spring came late and there was standing water in many fields, although there was less rainfall than usual. The warm weather in May helped the crops catch up, and the heading date was similar to the average. The weather was ideal for grain filling until the second half of June, after which the accumulation of assimilates was halted by two heat-waves. Ten to 14 extremely hot days caused forced ripening and shrivelled grains especially in late-maturing cultivars.

Among the biotic stress factors, considerable spontaneous infection with powdery mildew was observed in spring. The level of natural infection with leaf rust was lower than the previous year, but Fusarium head blight was reported in some areas. Harvest began relatively late, in mid-July, but the weather was dry and favorable. The yield average was moderate (4.06 t/ha) and a total of 4.3 x 10⁶ tons of good quality wheat was produced in Hungary. [p. 26]

Breeding. [p. 26-29]

Z. Bedö, L. Láng, O. Veisz, G. Vida, I. Karsai, M. Rakszegi, K. Mészáros, and D. Pribék.

The winter wheat cultivar Mv Kolo was registered in Hungary and Mv Vekni in Slovakia this year. Several cultivars already registered in Hungary were granted registration in foreign countries; Mv Suba in Serbia, Mv SŸveges in Croatia, and Mv Marsall in Romania.

Mv Kolo (Mv 417-03) is a medium, early maturing, top quality cultivar with good abiotic stress resistance, selected from the cross 'Mironovskaya-Ostistaya / Atay-85 // Alföld'. Yield level in official trials varied between 6 and 7.5 t/ha. The frost resistance level determined in phytotron tests is good, and it has above-average yield stability in dry years. Mv Kolo has 14.3-16.3% protein content and 33-36% wet-gluten content, with excellent gluten quality. The HWM-glutenin composition is 2*, 7+9, 5+10. Mv Kolo does not carry the T1B·1R translocation. Dough quality is outstanding, whether determined by Farinograph or Alveograph. Mv Kolo is characterized by a reliably high falling number. The cultivar is moderately resistant to powdery mildew, has good field resistance to leaf rust, and is resistant to the stem rust population used in the local artificially inoculated nursery.

Mv Vekni is a mid-late, hard red cultivar with a high gluten content. Good adaptability is proven by the ability to give high yields in production zones with widely differing climates. Mv Vekni has good winter hardiness and lodging resistance, allowing it to be reliably grown. Flour with a high water uptake can be ground from the grain. With a high gluten content (34-38%) and moderate falling number stability, bread-making quality is on par with that of Mv Magdaléna and Mv Csárdás, the cultivars used most widely for conventional bread-making purposes in Hungary. Mv Vekni has the T1B·1R translocation. The cultivar is moderately susceptible to powdery mildew and resistant to leaf rust and stem rust.

Wheat transformation. Experiments in wheat transformation were done with the biolistic method (gene gun) and Agrobacterium tumefaciens inoculation using the spring wheat cultivar Cadenza and the winter wheat cultivar Mv Emese, which have good adaptability, as the model plants.

A storage-protein gene from Amaranthus hypochondriacus (AMA1) and the 1Ax2*B storage-protein gene, isolated from the wheat cultivar Bánkúti 1201, were used as target genes in the biolistic transformation. Twenty experiments with the AMA1 gene, using ppt (herbicide) selection, involved a total of 720 embryos from Mv Emese and 1,500 from Cadenza. After the bombardment of the isolated embryos, nine surviving plants of Mv Emese and 37 of Cadenza were planted out into peat cubes. Seven of the Cadenza plants and four of the Mv Emese plants that survived selection were proved to be transgenic using PCR. For the 1Ax2*B gene, 270 embryos of each genotype were transformed, after which six herbicide-resistant plants of Cadenza and 11 of Mv Emese were grown. The DNA of these plants is currently being analyzed by PCR.

The presence of the transgene (bar or AMA1) also was detected in genomic DNA extracted from the leaves of T₁ plants raised from T₁ seeds produced on T₀ transgenic plants transformed in 2005. The expression of the mRNA extracted from immature seeds of these plants was confirmed using rtPCR. The expression of the protein coded by the AMA1 gene is now being investigated using Western blotting. Herbicide spraying was used to examine the expression of the herbicide-resistance gene. The expression of the protein coding for resistance was confirmed in plants from 12 independent transgenic lines.

Work has begun on the genetic transformation of immature embryos freshly isolated from the winter wheat Mv Emese and the spring wheat Cadenza using the hypervirulent A. tumefaciens strain AGL1 (pAL154/156). The T-DNA of the pGreen-based pAL156 plasmid contains the bar selection marker gene and a modified uidA (GUS) reporter gene, which includes an intron inhibiting gene expression in Agrobacterium. Gene transfer was detected using the histochemical reaction of GUS. In order to improve the efficiency of callus induction and plant regeneration, several hormone concentration combinations were tested. The in vitro selection of the first candidate transformants is now in progress. After selection, the presence of the GUS and bar genes incorporated into the plants will be detected with the help of specific primers.

Molecular markers. Molecular markers have been used to identify the Lr37 gene responsible for leaf rust resistance in a number of wheat genotypes . Among the wheat lines bred in Martonvásár, the gene complex Lr37-Yr17-Sr38 originating from source VPM-1 is carried by the registered cultivar Mv Vekni and the advanced lines Mv17-04 and Mv21-2000. The presence of the Lr19 gene has been demonstrated in genotypes Mv12-04, Mv35-06, Mv06-07, MVM28-04, and Mv14-06, whereas the Lr24 gene is in Mv Hombár, Mv08-03, and Mv15-06.

In order to discover molecular markers linked with the yellow index, which is closely correlated with the yellow pigment content of durum wheat, a bulk segregant analysis was made on 98 advanced lines from a progeny population developed using parents exhibiting large deviation for the yellow index and on the DNA of the parents. Five primers were used to identify nine polymorphic RAPD markers, and linkage between these markers and 2-year means of the yellow index was tested by means of correlation analysis. The coefficient of determination had values between 0 and 17%; the closest linkage being found for marker OPA16800. Multiple regression gave a coefficient of multiple determination of R² = 0.421 for all nine markers, and R² = 0.377 when only the four significant markers were considered. Discriminant analysis was used to check whether the marker data confirmed groupings made on the basis of the yellow index. When all nine markers were used, 80% of the lines were grouped correctly. Using only four markers, this value dropped to 77%, but the analysis confirmed 93.47% of the groupings of lines with low yellow index, indicating that lines with poor technological quality could be reliably identified using these markers.

Wheat quality. Genotypes over-expressing 1Bx7 HMW glutenin are bred in order to develop wheat with strong, elastic dough. Adaptable lines are selected from populations developed by crossing Glenlea, N93-326, and sublines of Bankuti 1201, which carry the gene responsible for Bx7 over-expression. On the basis of agronomic traits and farinograph curve stability, line Mv08-07 appears to be competitive.

Based on previous analyses that indicated the starch properties of normal winter wheat cultivar populations was unexpectedly variable and on the interest exhibited by the processing industry, tests were made to determine the starch content and amylose/amylopectin ratios of wheat genotypes, the viscosity of the starch suspension, and the level of starch damage. The starch content of the tested cultivars ranged from 60Ð88% and the amylose content from 18-34%. Values of 10.5-20.5 UCDc were found for the level of starch damage from wheat. When measuring RVA viscosity, greater differences were recorded when testing starch samples (4,706-6,310 cP final viscosity) than for flour samples (3,019-3,775 cP).

Disease-resistance studies. Powdery mildew isolates collected in the neighborhood of Martonvásár were used to determine the race composition of the pathogen population, the degree of virulence, and the effectiveness of known resistance genes. The following wheat powdery mildew races were dominant in 2006 (frequency in parentheses): 51 (26.7%), 72 and 76 (15.1%), and 77 (10.9%). The number of virulence genes in the pathogen population averaged 5.89. Almost complete protection against the wheat powdery mildew isolates tested was provided by the Pm4a+ resistance gene.

Fusarium head blight resistance of lines developed from populations of old Hungarian cultivars together with foreign sources of resistance was investigated in an artificially inoculated field nursery. The level of head blight severity for nine of the lines developed from populations of old Hungarian cultivars was less than 10%. Among the Martonvásár genotypes, a low level of FHB severity was recorded for Mv Emese, Mv Palotás, Mv Táltos, Mv Kolo, and line Mv08-05.

A survey was made of the virus composition of winter wheat, winter barley, durum wheat, winter oat and triticale crops. The wheat dwarf virus (WDV) was identified on almost 100% of plants exhibiting symptoms.

Abiotic stress resistance studies. As part of the research on abiotic stress resistance, changes in the antioxidant enzyme activity of six wheat cultivars grown in phytotron climatic chambers were studied spectrophotometrically in the course of 15-day heat stress treatment at the beginning of shooting and from the 12th day after heading. High temperature induced substantial differences in the activities of five antioxidant enzymes over the course of plant development. In response to heat stress, a decrease in the antioxidant enzyme activity was observed as the plants aged. High temperature had a degrading effect on the enzymes, manifested as a reduction in the level of activity. The greatest stability was observed for glutathione-S-transferase, catalase, and ascorbate peroxidase. The least change as the result of heat treatment was observed for glutathione reductase, whereas the most instable enzyme proved to be guaiacol peroxidase.

The effect of climatic components on the biomass and yield of cereals and on the quality of the grain yield was investigated in long-term experiments. The appearance and course of diseases was monitored, together with their effect on agronomic traits. For most of the cultivars tested, |the protein content of the grain was significantly lower in plants infected with leaf diseases than in those protected by spraying. The gluten quality (gluten index and gluten extension) suggested, in general, that in the case of natural infection the dough is firmer and less elastic than that of sprayed plants.

In a series of model experiments in the phytotron, the effects, interactions, and correlations of increases in mean temperature and atmospheric CO₂ concentration on the biomass production and yields of various cereal species and cultivars were investigated. High temperature generally accelerated plant development, leading to earlier heading and maturity, sometimes by as much as 5-13 days. Because of the shortening of the vegetation period, there was less biomass accumulation, with a consequent reduction in the grain number/plant and the yield quantity. In general, CO₂ fertilization had a positive influence on biomass accumulation and on quantitative yield parameters. The grain protein content of plants grown at higher atmospheric CO₂ concentration changed in some cultivars and remained constant in others.

Publications. [p. 29]

• Balla K, Bedö Z, and Veisz O. 2006. Effect of heat and drought stress on the photosynthetic processes of wheat. Cereal Res Commun 34(1):381-384.
  
• Bányai J, Szqcs P, Karsai I, Mészáros K, Kuti Cs, Láng L, and Bedö Z. 2006. Identification of winter wheat cultivars by simple sequence repeats (SSRs). Cereal Res Commun 34(2-3):865-870.
  
• Bencze Sz, Bedö Z, and Veisz O. 2006. Variation in the leaf composition of winter wehat varieties due to soil nitrogen content and elevated atmospheric CO2 level. Cereal Res Commun 34(1):401-404.
  
• Komáromi J, Vida Gy, Puskás K, Szunics L, and Veisz O. 2006. Identification of wheat genotypes with adult plant resistance to powdery mildew. Cereal Res Commun 34(2-3):1051-1058.
  
• Kuti Cs, Láng L, and Bedö Z. 2006. Pedigree records in plant breeding: from independent data to interdependent data structures. Cereal Res Commun 34(2-3):911-918.
  
• Pribék D, Pocsai E, Vida Gy, and Veisz O. 2006. Presence of wheat dwarf virus, cereal yellow dwarf virus-RPV and barley yellow dwarf viruses in cereal species in Martonvásár. Cereal Res Commun 34(1):625-628.
  
• Puskás K, Vida Gy, Komáromi J, Bürstmayr H, Lemmens M, Bedö Z, and Veisz O. 2006. Study of Fusarium head blight resistance in wheat microsatellite markers. Cereal Res Commun 34(1):629-632.
  
• Rakszegi M, Láng L, and Bedö Z. 2006. Importance of starch properties in quality oriented wheat breeding. In: Proc V Alps Adria Scientific Workshop, Opatija, Croatia, 6-11 March, 2006 (Hidvégi S, Ed). Cereal Res Commun 34(1):637-640.
  
• Uhrin A, Vida Gy, Gál M, Láng L, and Bedö Z. 2006. Marker-assisted selection for leaf rust resistance gene Lr37 in the Martonvásár breeding programme. In: Proc V Alps Adria Scientific Workshop, Opatija, Croatia, 6-11 March, 2006 (Hidvégi S, Ed). Cereal Res Commun 34(1):89-91.
  

Cell Biology Department. [p. 29-30]

B. Barnabás, M. Molnár-Láng, G. Linc, É. Szakács, K. Jäger, I. Molnár, F. Bakos, H. Ambrus, A. Schneider, A. Sepsi, and A. Fábián.

Induction of chromosome rearrangements in a 4H(4D) wheat-barley substitution using a wheat line containing a ph suppressor gene. Translocation lines were developed by inducing homoeologous chromosome pairing in a 4H(4D) wheat-barley substitution line previously developed in Martonvásár. We hoped to incorporate various segments of the barley 4H chromosome from the 4H(4D) substitution into wheat. Observations were made on the frequency with which wheatÐbarley translocations appeared in the F₂ progeny from a cross between the line CO4-1, which carries the Ph suppressor gene from Ae. speltoides and, thus, induces a high level of homoeologous chromosome pairing, and the 4H(4D) wheat-barley substitution line. Translocations were identified by means of genomic in situ hybridization. Of the 117 plants examined, three (2.4%) did not contain translocations. A total of four translocations were observed; one plant contained two different translocations. The translocations consisted of one centric fusion, two dicentric translocations, and one acrocentric chromosome. Plants with translocations were grown in the phytotron and selection of homozygous translocation lines was made in the F₃ progeny.

Characterization of chromosome-specific S-SAP markers and their use to study genetic diversity in Aegilops species. The short, interspersed nuclear element (SINE), Au, was used to develop sequence-specific amplified polymorphism (S-SAP) markers for the U- and M-genome chromosomes. The markers were localized using wheat-Ae. geniculata and wheat-Ae. biuncialis disomic chromosome addition lines. Thirty-seven markers distributed over six U- and six M-genome chromosomes were produced. Genetic diversity studies on 37 accessions of Ae. biuncialis, Ae. comosa, Ae. geniculata, and Ae. umbellulata suggested that Ae. biuncialis arose from its diploid ancestors more recently than Ae. geniculata. Several earlier studies indicated that the M genomes in polyploid Aegilops species had accumulated substantial rearrangements, whereas the U genomes remained essentially unmodified. However, this cannot be attributed to the preferential insertion of retroelements into the M-genome chromosomes. Fourteen markers from a total of eight chromosomes were sequenced. Three markers were similar to known plant genes. One marker was derived from an LTR-retrotransposon. Ten markers did not match to any known DNA sequences, suggesting that they were located in the highly variable intergenic regions.

Development and molecular cytogenetic identification of new winter wheat/winter barley (Martonvásári 9 kr1/Igri) disomic addition lines. A series of winter wheat/winter barley disomic addition lines were developed from hybrids between winter wheat line Martonvásári 9 kr1 and the German two-rowed winter barley cultivar Igri. The barley chromosomes in a wheat background were identified from the fluorescent in situ hybridization patterns obtained with various combinations of repetitive DNA probes GAA-HvT01 and pTa71-HvT01. The disomic addition lines 2H, 3H, and 4H, and the 1HS isochromosome were identified on the basis of a two-color FISH with the DNA probe pairs GAA-pAs1, GAA-HvT01 and pTa71-HvT01. Genomic in situ hybridization was used to confirm the presence of the barley chromosomes in the wheat genome. The identification of the barley chromosomes in the addition lines was further confirmed with SSR markers. The addition lines also were characterized morphologically.

Effect of heat stress and water deficit on reproductive development in wheat. The combined effect of elevated temperature and water deficit on the meiosis, flowering, and early seed development of the drought resistant wheat genotype Plainsman and sensitive Cappelle Desprez were studied. These processes lead to the development of the male and female gametes and those ensuring that the fusion of the gametes and the development of the embryo and endosperm should take place undisturbed. The effects of various kinds of abiotic stress on these processes are different, but in all cases negative, and their influence always results in a decline in the yield quantity. In maize, the effects of these stressors on reproductive development have been studied extensively, but a lack of information exists on their effect on some stages in the gametophytic development in wheat. High temperature and drought applied at the stage of gametogenesis resulted in the production of dysfunctional male and female gametophytes. At the time of flowering and early seed development, stressors negatively affected the fertility rates. The duration of dry matter accumulation decreased, so a dramatic reduction in 1,000-kernel weight was observed in both genotypes. The treatment did not affect the germination ability of seeds treated during the early grain-filling period, but the number of seminal roots markedly decreased in seedlings of the sensitive genotype.

Publications. [p. 30]

• Ascough G, Bakos F, Balázs E, Barnabás B, and van Staden J. 2006. Screening South African wheat germplasm for androgenic competence. S Afr J Bot 72:40-45.
  
• Bakos F, Darkó É, Pulido A, Géspér L, Ambrus H, Olmedilla A, and Barnabás B. 2006. Effect of aluminium treatment on the in vitro development of wheat microspores in anther and isolated microspore culture systems. In: From gametes to genes, Proc XIXth Internat Cong on Sexual Plant Reproduction (Barnabás B and Jáger K, Eds), Budapest, Hungary, 11-15 July. Pp. 116-117.
  
• Dudits D, Jáger K, Ötvös K, Ayaydin F, Kelemen Zs, Fábián A, Kristóf Z, Fehér A, and Barnabás B. 2006. Basic molecular mechanisms of cell cycle regulation and activation of wheat egg cells. In: From gametes to genes, Proc XIXth Internat Cong on Sexual Plant Reproduction (Barnabás B and Jäger K, Eds), Budapest, Hungary, 11-15 July. Pp. 17-18.
  
• Jäger K, Fábián A, and Barnabás B. 2006. Effect of heat stress and water deficit on embryo development in wheat (Triticum aestivum L.). In: From gametes to genes, Proc XIXth Internat Cong on Sexual Plant Reproduction (Barnabás B and Jäger K, Eds), Budapest, Hungary, 11-15 July. Pp. 74-75.
  
• Linc G, Molnár I, Schneider A, and Molnár-Láng M. 2006. Development of wheat-barley and wheat-Ae. biuncialis disomic additions using a doubled haploid technique, and identification of the lines by FISH. In: From gametes to genes, Proc XIXth Internat Cong on Sexual Plant Reproduction (Barnabás B and Jäger K, Eds), Budapest, Hungary, 11-15 July. Pp. 176-177.
  
• Molnár-Láng M, and Szakács É. 2006. Development and molecular cytogenetic identification of new winter wheat 'Martonvásári 9 kr1'/winter barley 'Igri' disomic addition lines. In: EWAC Newsletter (Börner A, Pankova K, and Snape JW, Eds). Pp. 26-30.
  
• Molnár-Láng M, Szakács É, and Linc G. 2006. Identification of newly developed wheat/winter barley addition lines using fluorescence in situ hybridization and SSR markers. In: Proc Vereinigung der PflanzenzŸchter und Saatgutkaufleute Österreichs HBLFA Raumberg-Gumpenstein, 21-23. November 2006. 57. Pp. 75-77.
  
• Molnár-Láng M, Szakács É, Linc G, and D Nagy E. 2006. Production and molecular cytogenetic identification of new winter wheat/winter barley disomic addition lines. In: Proc 7th Internat Wheat Conf, Mar del Plata, Argentina, 27. November- 4 December 2005 (in press).
  Szqcs A, Jurca M, Jäger K, Barnabás B, and Fehér A. 2006. Genomic approaches to reveal gene expression changes during fertilization and early seed development in wheat. In: From gametes to genes, Proc XIXth Internat Cong on Sexual Plant Reproduction (Barnabás B and Jäger K, Eds), Budapest, Hungary, 11-15 July. Pp. 65.

Genetic and physiological studies. [p. 31]

G. Galiba, G. Kocsy, A. Vágújfalvi, A. Bálint, F. Szira, A. Soltész, and T. Kellcs.

Effect of low temperature on gene expression. The frost tolerance locus Fr-Am2 was recently discovered in T. monococcum subsp. monococcum. Mapping data showed that 11 TmCbf genes are clustered at this locus in a 0.8 cM region. A novel mapping population is being developed to identify which of the Cbf genes are responsible for the differences in frost tolerance between the einkorn parental lines at the Fr-Am2 locus. The changes induced by cold or by chromosome 5A in the expression of wheat candidate genes selected by transcript profiling were confirmed by Northern analysis and RT-PCR.

Physiological changes induced by low temperature. The mechanism of the contribution of light during the development of freezing tolerance was investigated in winter wheat plants. Light induced the activity of certain antioxidant enzymes and altered the lipid composition and the metabolism of salicylic acid. Low-temperature hardening in the light caused a downshift in the far-red-induced AG thermoluminescence band. The faster dark re-reduction of P700+, monitored by 820-nm absorbance, could also be observed in these plants. These results suggest that the induction of cyclic photosynthetic electron flow may also contribute to the advantage of frost hardening under light conditions in wheat plants.

Drought tolerance studies. In order to determine QTL involved in osmotic and drought tolerance in barley, the Oregon Wolfe Barley (OWB) mapping population was examined at germination, as seedlings, and at maturity. Water stress was induced in young plants by adding polyethylene-glycol (PEG) to the growing solution. Limited watering was used at the mature stage. The most effective QTL for drought tolerance were found to be different in each developmental stage. The QTL influencing the yield parameters of mature plants under drought stress were different from those affecting osmotic adjustment in barley.

Publications. [p. 32]

• Apostol S, Szalai G, Sujbert L, Popova LP, and Janda T. 2006. Non-invasive monitoring of the light-induced cyclic photosynthetic electron flow during cold hardening in wheat leaves. Zeitschrift für Naturforsch C 61(9-10):734-740.
  
• Miller AK, Galiba G, and Dubcovsky J. 2006. A cluster of 11 CBF transcription factors is located at the frost tolerance locus Fr-Am2 in Triticum monococcum. Mol Genet Genomics 275(2):193-203.
  
• Szira F, Balint A, Galiba G, Rajeev KV, and Börner A. 2006. Mapping of QTLs for drought tolerance in barley at different developmental stages. Vorträge für Pflanzenzüchtung 70(1):113-115.

Cereal Genebank. [P. 31-32]

G. Kovács.

Characterization of cereal genetic resources. Last year, the agronomic performance and flowering of 120 new einkorn lines were tested, and an einkorn core collection established. Several new emmer genotypes and landraces were obtained via germ plasm exchange with other cereal genebanks and via collection from French farmers, who still cultivate them. Growth habit and winter hardiness were determined in these emmer wheats, and the frost-tolerant winter types were used in crosses to transfer their quality traits and frost tolerance to advanced winter durum lines. A new interspecific crossing program was started in order to transfer the biotic resistance of einkorn to other cultivated Triticum species. Einkorn lines with good crossability were identified in durum/einkorn crosses, and artificial primary hexaploid plants were produced with an ABA-genome structure. These fertile lines will be used for crossing with other hexaploid wheat species such as bread wheat, spelt wheat, and T. zhukovskyi.

Organic breeding. Some years ago, organic breeding programs were initiated for einkorn and emmer under certified organic conditions. During this period, several new crosses were produced, resulting in relatively high-yielding lines with good abiotic tolerance. Two of the best-performing lines are currently being tested for VCU, prior to being introduced into cultivation. These lines have very good agronomic performance under organic growth conditions. The einkorn genotype Mv Alkor has excellent wet gluten content, with good bread-making quality, but is an extremely soft-grained type. By contrast, the emmer line is a very hard-grained type with excellent pasta-making quality. The new advanced einkorn lines include several gluten-free variants, the lutein content of which is now being improved.

Publications. [P. 32]

• Kovacs G. 2006. The possible use of founder effect to produce locally adapted cereal varieties. In: Cereal Crop Diversity: Implications for production and products (Oestergard H and Fontaine L, Eds). ITAB, Paris, France. Pp. 68-70.