Fusarium Species Associated with Fusarium Head Blight in Hungarian Wheat Fields.

The species composition of the genus Fusarium associated with Fusarium head blight (FHB) in wheat fields of Hungary in the year 2019 was assessed. Symptomatic wheat heads were collected at 20 geographical locations representing different ecosystems. A total of 256 Fusarium strains were isolated and identified by partial sequences of the translation elongation factor 1-alpha gene and, where required, the second-largest subunit of the DNA-directed RNA polymerase gene. Overall, Fusarium graminearum (58.2%) proved to be the dominant species, followed by F. annulatum (formerly F. proliferatum) (17.2%) and F. verticillioides (7.4%). The presence of all other species, including F. culmorum, in the population was less than 5%. F. graminearum was identified as the main species associated with FHB at 14 sampling sites. Fumonisin-producing F. annulatum, primarily known as the pathogen of maize in Hungary, was detected nearly as frequently as F. graminearum at three locations and dominated at two other sites. F. poae was not found during the survey. F. vorosii, a species that is believed to be of Asian origin and was already found in Hungary in 2002, was identified at two locations.

Characterization of the Protein and Carbohydrate Related Quality Traits of a Large Set of Spelt Wheat Genotypes.

Spelt wheat (Triticum aestivum subsp. spelta L.) is an underexploited hexaploid wheat species that has become an increasingly fashionable raw material of bakery products in the last decades, partly because of its ability to grow under organic agricultural conditions and partly because of the growing number of people following the trend of having a healthy diet. However, due to its difficult threshing, most research on spelt seed is based on a very limited number of genotypes. Therefore, we determined the physical, compositional, and breadmaking quality traits of 90 spelt genotypes in order to highlight the variation of these properties and to identify possible genetic resources for spelt improvement. The thousand kernel weight of the spelt genotypes ranged between 23.2 and 49.7 g, the protein content between 12.1% and 22.2%, the gluten index between 0.7 and 98.8, the dough stability between 0.0 and 19.6 min, and the starch damage between 6.3 and 19.4 UCD value. The average values showed that spelt has higher protein and gluten contents but weaker dough strength and stability than common bread wheat. The starch pasting temperature was also higher in spelt, but the starch damage was lower, resulting in lower water absorption. Some genebank accessions (MVGB142, 145, 353, and 525) and internationally available cultivars (Bohemia, Bodensonne, Black-Bearded, and White-Beardless) were identified as good genetic resources for improving the breadmaking-quality traits of spelt.

Variation in the Content and Composition of Tocols in a Wheat Population.

Wheat is a well-known source of B vitamins but also contains significant amounts of vitamin E and related tocols, which have a number of positive health benefits. However, there are no reports on increasing the tocol content of wheat. A prerequisite for increasing the tocol content is the identification of variation in its amount within wheat and related cereals. We therefore determined the tocol content and composition in the grain of 230 recombinant inbred lines (RILs) of a diverse biparental wheat population (Mv Toborzó/Tommi), showing variation in the total content from 13.69 to 45.18 µg/g d.m. The total content also showed transgressive segregation in the population. The effect of the genotype on the variance components of tocols was studied, and the broad-sense heritability was calculated to be 0.71. The lines were also grouped based on their tocol content and analyzed for their chemical composition and breadmaking quality. The high heritability value and the wide variation found in the total amount indicate that increasing the content of tocols is a possible breeding strategy.

Investigation of Protein and Epitope Characteristics of Oats and Its Implications for Celiac Disease.

The use of pure oats (oats cultivated with special care to avoid gluten contamination from wheat, rye, and barley) in the gluten-free diet (GFD) represents important nutritional benefits for the celiac consumer. However, emerging evidence suggests that some oat cultivars may contain wheat gliadin analog polypeptides. Consequently, it is necessary to screen oats in terms of protein and epitope composition to be able to select safe varieties for gluten-free applications. The overall aim of our study is to investigate the variability of oat protein composition directly related to health-related and techno-functional properties. Elements of an oat sample population representing 162 cultivated varieties from 20 countries and the protein composition of resulting samples have been characterized. Size distribution of the total protein extracts has been analyzed by size exclusion-high performance liquid chromatography (SE-HPLC) while the 70% ethanol-extracted proteins were analyzed by RP-HPLC. Protein extracts separated into three main groups of fractions on the SE-HPLC column: polymeric proteins, avenins (both containing three subgroups based on their size), and soluble proteins, representing respectively 68.79-86.60, 8.86-27.72, and 2.89-11.85% of the total protein content. The ratio of polymeric to monomeric proteins varied between 1.37 and 3.73. Seventy-six reversed phase-HPLC-separated peaks have been differentiated from the ethanol extractable proteins of the entire population. Their distribution among the cultivars varied significantly, 6-23 peaks per cultivar. The number of appearances of peaks also showed large variation: one peak has been found in 107 samples, while 15 peaks have been identified, which appeared in less than five cultivars. An estimation method for ranking the avenin-epitope content of the samples has been developed by using MS spectrometric data of collected RP-HPLC peaks and bioinformatics methods. Using ELISA methodology with the R5 antibody, a high number of the investigated samples were found to be contaminated with wheat, barley, or rye.

Abiotic Stress Response of Near-Isogenic Spring Durum Wheat Lines under Different Sowing Densities.

A detailed study was made of changes in the plant development, morphology, physiology and yield biology of near-isogenic lines of spring durum wheat sown in the field with different plant densities in two consecutive years (2013-2014). An analysis was made of the drought tolerance of isogenic lines selected for yield QTLs (QYld.idw-2B and QYld.idw-3B), and the presence of QTL effects was examined in spring sowings. Comparisons were made of the traits of the isogenic pairs QYld.idw-3B++ and QYld.idw-3B-- both within and between the pairs. Changes in the polyamine content, antioxidant enzyme activity, chlorophyll content of the flag leaf and the normalized difference vegetation index (NDVI) of the plot were monitored in response to drought stress, and the relationship between these components and the yield was analyzed. In the case of moderate stress, differences between the NIL++ and NIL-- pairs appeared in the early dough stage, indicating that the QYld.idw-3B++ QTL region was able to maintain photosynthetic activity for a longer period, resulting in greater grain number and grain weight at the end of the growing period. The chlorophyll content of the flag leaf in phenophases Z77 and Z83 was significantly correlated with the grain number and grain weight of the main spike. The grain yield was greatly influenced by the treatment, while the genotype had a significant effect on the thousand-kernel weight and on the grain number and grain weight of the main spike. When the lines were compared in the non-irrigated treatment, significantly more grains and significantly higher grain weight were observed in the main spike in NIL++ lines, confirming the theory that the higher yields of the QYld.idw-3B++ lines when sown in spring and exposed to drought stress could be attributed to the positive effect of the "Kofa" QTL on chromosome 3B.

Effect of Multi-Year Environmental and Meteorological Factors on the Quality Traits of Winter Durum Wheat.

A detailed study was made of the effect of rainfall, average temperature and hot days on the gluten index and Minolta b* value of winter durum wheat sown in the field in 16 consecutive crop years (2005-2020). The joint analysis of these two technological quality traits represented a complex (plant-environment-meteorological factors) approach for the identification of durum wheat cultivars carrying an optimum combination of the two traits and for the determination of quality stability. The results of GGE-biplot analysis indicated that the cultivar that had the most favorable combination of the traits was 'MVP', while cultivar 'GKS' had the best gluten strength and 'MVH' the best yellow pigment content. Correlation analysis and stepwise regression between various meteorological factors (rainfall, mean temperature, number of heat days per 10-day period during grain-filling) and the two technological quality traits indicated that the expected value of the quality traits could be reliably estimated based on meteorological factors, with a generally negative effect on gluten index and a positive one on yellowness in all cultivars.

A novel approach to the characterization of old wheat (Triticum aestivum L.) varieties by complex rheological analysis.

BACKGROUND: Lines of the internationally recognized old Hungarian Bánkúti 1201 variety are important genetic resources for breeding programmes. Their protein composition and gluten dependent technological traits have been comprehensively studied, however, little information is available about their carbohydrate dependent viscous properties. The aim of this work was to obtain comprehensive rheological characterization of all sublines of Bánkúti 1201 maintained at Martonvásár and to investigate their variability if the carbohydrate dependent viscous behaviour was also included in the analyses. RESULTS: The majority of the lines reflected the famously good mixing quality of Bánkúti, however, much higher diversity of pasting behaviour was detected. Cluster analysis of the Mixolab data was performed resulting in four sample groups. Since several lines of similar mixing properties had significantly different pasting characteristics, it was assumed that classification was mainly based on the viscous properties. From each cluster two to three representative samples were selected for wider examination using conventional testing methods. These results also supported the higher variability of pasting behaviour of the lines, which can be critical for end product quality. The members of the second cluster can be highlighted due to their waxy wheat like behaviour. CONCLUSIONS: Possible reasons for the great variability of pasting behaviour could be the compositional and structural differences of starch and other carbohydrates (e.g. arabinoxylans). Complex rheological characterization and study of molecular background can provide information about important traits from the point of view of technology and product development, which are unknown in the case of old wheat varieties and landraces. © 2020 Society of Chemical Industry.

Stability analysis of wheat lines with increased level of arabinoxylan.

Plant breeders have long sought to develop lines that combine outstanding performance with high and stable quality in different environments. The high-arabinoxylan (AX) Chinese variety Yumai-34 was crossed with three Central European wheat varieties (Lupus, Mv-Mambo, Ukrainka) and 31 selected high-AX lines were compared for physical (hectolitre weight, thousand grain weight, flour yield), compositional (protein content, gluten content, pentosan) and processing quality traits (gluten index, Zeleny sedimentation, Farinograph parameters) in a three-year experiment (2013-2015) in the F7-F9 generations. The stability and heritability of different traits, including the relative effects of the genotype (G) and environment (E), were determined focusing on grain composition. The contents of total and water-soluble pentosans were significantly affected by G, E and G × E interactions, but the heritability of total (TOT)-pentosan was significantly lower (0.341) than that of water-extractable (WE)-pentosan (0.825). The main component of the pentosans, the amount and composition (arabinose:xylose ratio) of the arabinoxylan (AX), was primarily determined by the environment and, accordingly, the broader heritability of these parameters were 0.516 and 0.772. However, genotype significantly affected the amount of water-soluble arabinoxylan and its composition and thus the heritability of these traits was also significant (0.840 and 0.721). The genotypes exhibiting higher stability of content of TOT-pentosan also showed more stable contents of WE-pentosan. There was a positive correlation between the stability of contents of WE-pentosan and WE-AX, while the stability of the WE-AX content and AX composition were also strongly correlated. Water absorption was strongly genetically determined with a heritability of 0.829 with the genotype determining 38.67% of the total variance. Many lines were grouped in the GGE biplot, indicating that they did not significantly differ stability.

Multiple Fungal Metabolites Including Mycotoxins in Naturally Infected and Fusarium-Inoculated Wheat Samples.

In this study, the occurrence of multiple fungal metabolites including mycotoxins was determined in four different winter wheat varieties in a field experiment in Croatia. One group was naturally infected, while the second group was inoculated with a Fusarium graminearum and F. culmorum mixture to simulate a worst-case infection scenario. Data on the multiple fungal metabolites including mycotoxins were acquired with liquid chromatography with mass spectrometry (LC-MS/MS) multi-(myco)toxin method. In total, 36 different fungal metabolites were quantified in this study: the Fusarium mycotoxins deoxynivalenol (DON), DON-3-glucoside (D3G), 3-acetyldeoxynivalenol (3-ADON), culmorin (CULM), 15-hydroxyculmorin, 5-hydroxyculmorin, aurofusarin, rubrofusarin, enniatin (Enn) A, Enn A1, Enn B, Enn B1, Enn B2, Enn B3, fumonisin B1, fumonisin B2, chrysogin, zearalenone (ZEN), moniliformin (MON), nivalenol (NIV), siccanol, equisetin, beauvericin (BEA), and antibiotic Y; the Alternaria mycotoxins alternariol, alternariolmethylether, altersetin, infectopyron, tentoxin, tenuazonic acid; the Aspergillus mycotoxin kojic acid; unspecific metabolites butenolid, brevianamid F, cyclo(L-Pro-L-Tyr), cyclo(L-Pro-L-Val), and tryptophol. The most abundant mycotoxins in the inoculated and naturally contaminated samples, respectively, were found to occur at the following average concentrations: DON (19,122/1504 µg/kg), CULM (6109/1010 µg/kg), 15-hydroxyculmorin (56,022/1301 µg/kg), 5-hydroxyculmorin (21,219/863 µg/kg), aurofusarin (43,496/1266 µg/kg). Compared to naturally-infected samples, Fusarium inoculations at the flowering stage increased the concentrations of all Fusarium mycotoxins, except enniatins and siccanol in Ficko, the Aspergillus metabolite kojic acid, the Alternaria mycotoxin altersetin, and unspecific metabolites brevianamid F, butenolid, cyclo(L-Pro-L-Tyr), and cyclo(L-Pro-L-Val). In contrast to these findings, because of possible antagonistic actions, Fusarium inoculation decreased the concentrations of the Alternaria toxins alternariol, alternariolmethylether, infectopyron, tentoxin, tenuazonic acid, as well as the concentration of the nonspecific metabolite tryptophol.

1RS arm of Secale cereanum 'Kriszta' confers resistance to stripe rust, improved yield components and high arabinoxylan content in wheat.

Wheat-rye T1BL.1RS translocation is widespread worldwide as the genes on 1RS arm have positive effect on stress resistance, grain yield and adaptation ability of wheat. Nowadays, the T1BL.1RS wheat cultivars have become susceptible to rust diseases because of the monophyletic ('Petkus') origin of 1RS. Here we report and discuss the production and detailed investigation of a new T1BL.1RS translocation line carrying 1RS with widened genetic base originating from Secale cereanum. Line '179' exhibited improved spike morphology traits, resistance against stripe rust and leaf rust, as well as higher tillering capacity, fertility and dietary fiber (arabynoxylan) content than the parental wheat genotype. Comparative analyses based on molecular cytogenetic methods and molecular (SSR and DArTseq) makers indicate that the 1RS arm of line '179' is a recombinant of S. cereale and S. strictum homologues, and approximately 16% of its loci were different from that of 'Petkus' origin. 162 (69.5%) 1RS-specific markers were associated with genes, including 10 markers with putative disease resistance functions and LRR domains found on the subtelomeric or pericentromeric regions of 1RS. Line '179' will facilitate the map-based cloning of the resistance genes, and it can contribute to healthy eating and a more cost-efficient wheat production.

Rising atmospheric CO2 concentration may imply higher risk of Fusarium mycotoxin contamination of wheat grains.

Increasing atmospheric CO2 concentration not only has a direct impact on plants but also affects plant-pathogen interactions. Due to economic and health-related problems, special concern was given thus in the present work to the effect of elevated CO2 (750 µmol mol-1) level on the Fusarium culmorum infection and mycotoxin contamination of wheat. Despite the fact that disease severity was found to be not or little affected by elevated CO2 in most varieties, as the spread of Fusarium increased only in one variety, spike grain number and/or grain weight decreased significantly at elevated CO2 in all the varieties, indicating that Fusarium infection generally had a more dramatic impact on the grain yield at elevated CO2 than at the ambient level. Likewise, grain deoxynivalenol (DON) content was usually considerably higher at elevated CO2 than at the ambient level in the single-floret inoculation treatment, suggesting that the toxin content is not in direct relation to the level of Fusarium infection. In the whole-spike inoculation, DON production did not change, decreased or increased depending on the variety × experiment interaction. Cooler (18 °C) conditions delayed rachis penetration while 20 °C maximum temperature caused striking increases in the mycotoxin contents, resulting in extremely high DON values and also in a dramatic triggering of the grain zearalenone contamination at elevated CO2. The results indicate that future environmental conditions, such as rising CO2 levels, may increase the threat of grain mycotoxin contamination.

Powdery Mildew Resistance in Wheat Cultivar Mv Hombár is Conferred by a New Gene, PmHo.

A new powdery mildew resistance gene designated as PmHo was identified in 'Mv Hombár' winter wheat, bred in Martonvásár, Hungary. It has exhibited a high level of resistance over the last two decades. Genetic mapping of recombinant inbred lines derived from the cross 'Ukrainka'/Mv Hombár located this gene on chromosome 2AL. The segregation ratio and consistent effect in all environments indicated that PmHo is a major dominant powdery mildew resistance gene. The race-specific nature of resistance in Mv Hombár was shown by the emergence of a single virulent pathotype designated as 51-Ho. This pathotype was, to some extent, able to infect Mv Hombár, developing visible symptoms with sporulating colonies. Microscopic studies revealed that, in incompatible interactions, posthaustorial hypersensitivity reaction was the most prevalent but not exclusive plant defense response in Mv Hombár, and fungal growth was mostly arrested during haustorium formation or in the early stages of colony development. The delayed fungal development of the virulent pathotype 51-Ho may be explained by additional effects of other loci that were also involved in the powdery mildew resistance of Mv Hombár.

Dataset on the mean, standard deviation, broad-sense heritability and stability of wheat quality bred in three different ways and grown under organic and low-input conventional systems.

An assessment was previously made of the effects of organic and low-input field management systems on the physical, grain compositional and processing quality of wheat and on the performance of varieties developed using different breeding methods ("Comparison of quality parameters of wheat varieties with different breeding origin under organic and low-input conventional conditions" [1]). Here, accompanying data are provided on the performance and stability analysis of the genotypes using the coefficient of variation and the 'ranking' and 'which-won-where' plots of GGE biplot analysis for the most important quality traits. Broad-sense heritability was also evaluated and is given for the most important physical and quality properties of the seed in organic and low-input management systems, while mean values and standard deviation of the studied properties are presented separately for organic and low-input fields.

McGISH identification and phenotypic description of leaf rust and yellow rust resistant partial amphiploids originating from a wheat × Thinopyrum synthetic hybrid cross.

A Thinopyrum intermedium × Thinopyrum ponticum synthetic hybrid wheatgrass is an excellent source of leaf and stem rust resistance produced by N.V.Tsitsin. Wheat line Mv9kr1 was crossed with this hybrid (Agropyron glael) in Hungary in order to transfer its advantageous agronomic traits into wheat. As the wheat parent was susceptible to leaf rust, the transfer of resistance was easily recognizable in the progenies. Three different partial amphiploid lines with leaf rust resistance were selected from the wheat/Thinopyrum hybrid derivatives by multicolour genomic in situ hybridization. Chromosome counting on the partial amphiploids revealed 58 chromosomes (18 wheatgrass) in line 194, 56 (14 wheatgrass) in line 195 and 54 (12 wheatgrass) in line 196. The wheat chromosomes present in these lines were identified and the wheatgrass chromosomes were characterized by fluorescence in situ hybridization using the repetitive DNA probes Afa-family, pSc119.2 and pTa71. The 3D wheat chromosome was missing from the lines. Molecular marker analysis showed the presence of the Lr24 leaf rust resistance gene in lines 195 and 196. The morphological traits were evaluated in the field during two consecutive seasons in two different locations.

New Insights into the Life Cycle of the Wheat Powdery Mildew: Direct Observation of Ascosporic Infection in Blumeria graminis f. sp. tritici.

Although Blumeria graminis is an intensively studied pathogen, an important part of its life cycle (namely, the way ascospores initiate primary infections on cereal leaves) has not yet been explored in detail. This study reports, for the first time, the direct observation of this process in B. graminis f. sp. tritici using light and confocal laser-scanning microscopy. All the germinated ascospores produced a single germ tube type both in vitro and on host plant surfaces; therefore, the ascosporic and conidial germination patterns are markedly different in this fungus, in contrast to other powdery mildews. Germinated ascospores penetrated the epidermal cells of wheat leaves and produced haustoria as known in the case of conidial infections. This work confirmed earlier studies reporting that B. graminis chasmothecia collected from the field do not contain mature ascospores, only asci filled with protoplasm; ascospore development is induced by moist conditions and is a fast process compared with other powdery mildews. Although ascosporic infections are frequent in B. graminis f. sp. tritici in the field, as shown by this study and other works as well, a recent analysis of the genomes of four isolates revealed the signs of clonal or near-clonal reproduction. Therefore, chasmothecia and ascospores are probably more important as oversummering structures than genetic recombination factors in the life cycle of this pathogen.

A consensus framework map of durum wheat (Triticum durum Desf.) suitable for linkage disequilibrium analysis and genome-wide association mapping.

BACKGROUND: Durum wheat (Triticum durum Desf.) is a tetraploid cereal grown in the medium to low-precipitation areas of the Mediterranean Basin, North America and South-West Asia. Genomics applications in durum wheat have the potential to boost exploitation of genetic resources and to advance understanding of the genetics of important complex traits (e.g. resilience to environmental and biotic stresses). A dense and accurate consensus map specific for T. durum will greatly facilitate genetic mapping, functional genomics and marker-assisted improvement. RESULTS: High quality genotypic data from six core recombinant inbred line populations were used to obtain a consensus framework map of 598 simple sequence repeats (SSR) and Diversity Array Technology® (DArT) anchor markers (common across populations). Interpolation of unique markers from 14 maps allowed us to position a total of 2,575 markers in a consensus map of 2,463 cM. The T. durum A and B genomes were covered in their near totality based on the reference SSR hexaploid wheat map. The consensus locus order compared to those of the single component maps showed good correspondence, (average Spearman's rank correlation rho ρ value of 0.96). Differences in marker order and local recombination rate were observed between the durum and hexaploid wheat consensus maps. The consensus map was used to carry out a whole-genome search for genetic differentiation signatures and association to heading date in a panel of 183 accessions adapted to the Mediterranean areas. Linkage disequilibrium was found to decay below the r2 threshold=0.3 within 2.20 cM, on average. Strong molecular differentiations among sub-populations were mapped to 87 chromosome regions. A genome-wide association scan for heading date from 27 field trials in the Mediterranean Basin and in Mexico yielded 50 chromosome regions with evidences of association in multiple environments. CONCLUSIONS: The consensus map presented here was used as a reference for genetic diversity and mapping analyses in T. durum, providing nearly complete genome coverage and even marker density. Markers previously mapped in hexaploid wheat constitute a strong link between the two species. The consensus map provides the basis for high-density single nucleotide polymorphic (SNP) marker implementation in durum wheat.

Mapping of quantitative adult plant field resistance to leaf rust and stripe rust in two European winter wheat populations reveals co-location of three QTL conferring resistance to both rust pathogens.

KEY MESSAGE: We detected several, most likely novel QTL for adult plant resistance to rusts. Notably three QTL improved resistance to leaf rust and stripe rust simultaneously indicating broad spectrum resistance QTL. The rusts of wheat (Puccinia spp.) are destructive fungal wheat diseases. The deployment of resistant cultivars plays a central role in integrated rust disease management. Durability of resistance would be preferred, but is difficult to analyse. The Austrian winter wheat cultivar Capo was released in the 1989 and grown on a large acreage during more than two decades and maintained a good level of quantitative leaf rust and stripe rust resistance. Two bi-parental mapping populations: Capo × Arina and Capo × Furore were tested in multiple environments for severity of leaf rust and stripe rust at the adult plant stage in replicated field experiments. Quantitative trait loci associated with leaf rust and stripe rust severity were mapped using DArT and SSR markers. Five QTL were detected in multiple environments associated with resistance to leaf rust designated as QLr.ifa-2AL, QLr.ifa-2BL, QLr.ifa-2BS, QLr.ifa-3BS, and QLr.ifa-5BL, and five for resistance to stripe rust QYr.ifa-2AL, QYr.ifa-2BL, QYr.ifa-3AS, QYr.ifa-3BS, and QYr.ifa-5A. For all QTL apart from two (QYr.ifa-3AS, QLr.ifa-5BL) Capo contributed the resistance improving allele. The leaf rust and stripe rust resistance QTL on 2AL, 2BL and 3BS mapped to the same chromosome positions, indicating either closely linked genes or pleiotropic gene action. These three multiple disease resistance QTL (QLr.ifa-2AL/QYr.ifa-2AL, QLr.ifa.2BL/QYr.ifa-2BL, QLr.ifa-3BS/QYr.ifa.3BS) potentially contribute novel resistance sources for stripe rust and leaf rust. The long-lasting resistance of Capo apparently rests upon a combination of several genes. The described germplasm, QTL and markers are applicable for simultaneous resistance improvement against leaf rust and stripe rust.

Verification of STS markers for leaf rust resistance genes of wheat by seven European laboratories.

A set of Thatcher near-isogenic lines and two breeding lines were used to examine sequence tagged site (STS) markers linked to leaf rust resistance genes Lr9, Lr10, Lr19, Lr24, Lr28, Lr29, Lr35, and a simple sequenced repeat (SSR) marker for Lr39. The selected STS markers for resistance genes Lr9, Lr10, Lr19, Lr24 and Lr28 were identified in seven accessions by seven European laboratories. Near-isogenic lines of the spring wheat Thatcher were used as positive controls. Markers for resistance genes Lr9, Lr10, Lr19, Lr24 were identified in all seven laboratories as amplification products of 1100 bp, 310 bp, 130 bp and 310 bp, respectively. The STS markers linked to resistance genes Lr9, Lr10, Lr19, Lr24, Lr29, Lr35 and the SSR marker for Lr39 were robust and highly specific for these genes and will be useful in marker-assisted selection in wheat. However, the amplification product of 378 bp that corresponded with resistance gene Lr28 was detected in all accessions including genotypes lacking this gene in all seven laboratories. This marker needs to be improved.