Allelic Variation Analysis at the Vernalization Response and Photoperiod Genes in Russian Wheat Varieties Identified Two Novel Alleles of Vrn-B3

Heading time is an important agronomic trait affecting the adaptability and productivity of common wheat. In this study, 95 common wheat varieties from Russia and the late-maturing breeding line ‘Velut’ were tested for allelic diversity of genes having the strongest effect on heading. In this research, allelic variation at the Ppd-D1, Vrn-A1, Vrn-B1, Vrn-D1, and Vrn-B3 loci was tested. The Vrn-B1 and Vrn-B3 loci provided the largest contribution to genetic diversity. We found two novel allelic variants of the Vrn-B3 gene in the studied varieties. Ten varieties carried a 160 bp insertion in the promoter region, and the breeding line ‘Velut’ carried a 1617 bp insertion. These alleles were designated Vrn-B3e and Vrn-B3d, respectively. The analysis of the sequences showed the recent insertion of a retrotransposon homologous to the LTR retrotransposon (RLX_Hvul_Dacia_ RND-1) in the Vrn-B3d allele. Plants with the Vrn-B3e and the ‘Velut’ line with the Vrn-B3d allele headed later than the plants with the wild-type allele; among these plants, ‘Velut’ is the latest maturing wheat variety. Analysis of the gene expression of two groups of lines differing by the Vrn-B3 alleles (Vrn-B3d or vrn-B3) from the F2 population with ‘Velut’ as a parental line did not reveal a significant difference in the expression level between the groups. Additional research is required to study the reasons for the late maturation of the ‘Velut’ line. However, the studied wheat varieties could be used as a potential source of natural variation in genes controlling heading times.

Variability of selected quantitative traits in new spring barley genotypes

The research included 19 breeding lines and 4 cultivars of spring barley from the preliminary field experiments harvested in 2020 in Radzików. All barley samples were characterized for the content of protein, non-starch polysaccharides (NSP) with soluble (S-NSP) and insoluble (I-NSP) fractions and β-glucan. Additionally, viscosity of water extracts (WEV) was measured to determine the functional properties of the grain. It was the most diverse parameter (CV = 27%) and was significantly correlated with β-glucan content (r = 0.50; for p <0.05). This dependence is shown by the results obtained for the grain of the Avatar cultivar and the RAH 744/19 breeding line, in which the content of β-glucan (5.3% and 4.8%, respectively), as well as the WEV (3.3 mPa.s and 3.0 mPa.s, respectively) were the highest. The lowest content of β – glucan (3.5%) and one of the lowest WEV values (1.4 mPa.s) were observed for KWS Jessie cultivar. Principal component analysis (PCA) showed a substantial impact of the two components PC1 and PC2 on the variability of the analyzed material showing significant variability of the 5 barley genotypes and confirmed the previous results of biochemical analyzes. Our results made it possible to indicate several genotypes that may constitute a source of variability in breeding works aimed at improving the quality of barley. Presented study also show that the grain of some new barley genotypes, with a favorable chemical composition from a fodder and brewing perspective, is a good material for future use in industry.

Reactions of winter wheat, spring wheat, and barley cultivars to mechanical inoculation with wheat streak mosaic virus

Wheat streak mosaic virus (WSMV) causes sporadic epidemics in Montana which can threaten profitability of the state’s small grains production. One challenge for WSMV management in Montana is that most commercially available wheat and barley cultivars are susceptible to WSMV or their performance under WSMV pressure is unknown. In a three-year field study from 2017 to 2019 winter wheat, spring wheat, and barley cultivars were evaluated for their susceptibility to WSMV and yield performance under WSMV pressure. Plants were mechanically inoculated and WSMV incidence was assessed using DAS-ELISA. There was effective resistance to WSMV in breeding line CO12D922, which had consistently low WSMV incidence, highlighting promising efforts in the development of WSMV-resistant winter wheat cultivars. Moderate WSMV incidence and minor yield losses were observed from WSMV infection of commercial winter wheat ‘Brawl CL Plus’ and MSU breeding line MTV1681. Spring wheat cultivars in this study had high WSMV incidence of up to 100 % in ‘Duclair,’ ‘Egan,’ and ‘McNeal.’ High WSMV incidence was associated with severe yield losses as high as 85 % for Duclair and ‘WB9879CL’ in 2019, demonstrating a high degree of susceptibility to WSMV inoculation. Barley cultivars had considerably lower WSMV incidence compared to spring and winter wheat. Grain yield response to WSMV inoculation was variable between barley cultivars. The study provided an experimental basis for cultivar recommendations for high WSMV pressure environments and identified breeding lines and cultivars with potential resistance traits of interest to breeding programs that aim to develop WSMV-resistant cultivars.

Identification of Gray Leaf Spot Disease Candidate Gene in Narrow-Leafed Lupin (Lupinus angustifolius L.)

Selection for resistance against gray leaf spot (GLS) is a major objective in the lupin breeding programs. A segregation ratio of 1:1 (resistant:susceptible) in F8 recombinant inbred lines (RIL8) derived from a cross between a breeding line 83A:476 (resistant to GLS) and a wild accession P27255 (susceptible to GLS) indicated that GLS was controlled by a single major gene. To develop molecular markers linked to GLS, in the beginning, only 11 resistant lines and six susceptible lines from the 83A:476 and P27255 population were genotyped with MFLP markers, and three MFLP markers were identified to be co-segregated with GLS. This method was very efficient, but the markers were located outside of the gene, and could not be used in other germplasms. Then QTL analysis and fine mapping were conducted to identify the gene. Finally, the gene was narrowed down to a 241-kb region containing two disease resistance genes. To further identify the candidate gene, DNA variants between accessions Tanjil (resistant to GLS) and Unicrop (susceptible to GLS) were analyzed. The results indicated that only one SNP was detected in the 241 kb region. This SNP was located in the TMV resistance protein N-like gene region and also identified between 83A:476 and P27255. Genotyping the Tanjil/Unicrop RIL8 population showed that this SNP co-segregated with GLS resistance. The phylogenetic tree analysis of this gene among 18 lupin accessions indicates that Australian resistant breeding line/varieties were clustered into one group and carry two resistant alleles, while susceptible accessions were clustered into different groups.

Characterization of the Powdery Mildew Resistance Gene in Wheat Breeding Line KN0816 and its Evaluation in Marker-Assisted Selection

Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a destructive disease seriously threatening yield and quality of common wheat (Triticum aestivum L., 2n=6x=42, AABBDD). Characterization of resistance genes against powdery mildew is useful in parental selection and for developing disease resistant cultivars. Chinese wheat breeding line KN0816 has superior agronomic performance and resistance to powdery mildew at all growth stages. Genetic analysis using populations of KN0816 crossed with different susceptible parents indicated that a single dominant gene, tentatively designated PmKN0816, conferred seedling resistance to different Bgt isolates. Using a bulked segregant analysis (BSA), PmKN0816 was mapped to the Pm6 interval on chromosome arm 2BL using polymorphic markers linked to the catalogued genes Pm6, Pm52, and Pm64, and flanked by markers CISSR02g-6 and CIT02g-2 both with genetic distances of 0.7 cM. Analysis of closely linked molecular markers indicated that the marker alleles of PmKN0816 differed from those of other powdery mildew resistance genes on 2BL, including Pm6, Pm33, Pm51, Pm64, and PmQ. Based on the genetic and physical locations and response pattern to different Bgt isolates, PmKN0816 is most likely a new powdery mildew resistance gene and confers effective resistance to all the 14 tested Bgt isolates. In view of the elite agronomic performance of KN0816 combined with the resistance, PmKN0816 is expected to become a valuable resistance gene in wheat breeding. To transfer PmKN0816 to different genetic backgrounds using marker-assisted selection (MAS), closely linked markers of PmKN0816 were evaluated and four of them (CIT02g-2, CISSR02g-6, CIT02g-10, and CIT02g-17) were confirmed to be applicable for MAS in different genetic backgrounds.

Environmental trial of pea breeding acessions

The aim of the study was to determine the information content and adequacy of the ecological testing points of the pea breeding material and to identify the "ideal" genotype. Materials and methods. The breeding material in the experiments was represented by cultivars bred at the PPI nd.a. V.Ya. Yuriev (Tsarevych, Oplot, Otaman, Metsenat, Korvet, Haiduk, and Malakhit) and ten breeding lines (SL 11-129, SL 11-213, SL 11-55, SL 11-58, SL 10-37, SL 11-32, SL 10-132, SL 09-118, SL 11-166, and SL 11-176). In addition there was one cultivar (Svit) bred at the Plant Breeding and Genetics Institute. All the cultivars are leafless, semi-dwarf, mid-ripening, except for Tsarevych (mid-early). The field experiments were carried out in accordance with the methods of field experimentation, using the conventional pea growing technology. The seeding rate was 1.2 million germinable seeds/ha; the plot area was 10 m2. To evaluate the accessions for the variability in different environments, we used a regression model developed by S.A. Eberhart and W.A. Russel, where the regression coefficient is an indicator of the genotype-environment interaction. This model is included in "Guidelines for Environmental Trials of Corn". Results and discussion. Thus, comparing the regression coefficient in pea cultivars Oplot, Tsarevych, Haiduk, Korvet, and Metsenat, we could conclude that these accessions were highly intensive in the OSES conditions and extensive in the PPI NAAS conditions (except for Metsenat). Regarding the regression coefficient in the breeding lines, none of them had a regression coefficient of 1.0. Over the study period, the regression coefficient was 1.4 only in line SL 11-58 (PPI NAAS) and 1.2 (OSES), characterizing this line as intensive regardless of the place of cultivation. Taking into account that the regression coefficient values of <1 are intrinsic to extensive accessions, lines SL 10-132 (RC = 0.4) and SL 11-176 (RC = 0.8) are preferred. Because these accessions also have a high genotypic effect. In addition, the regression coefficient in breeding line SL 09-118 was 0.9, with a genotypic effect of 0.07. Such combination of the indicators characterizes the line as relatively stable, with sufficient potential performance, and this breeding line will not be demanding to growing conditions similar to the OSES ones. Conclusions. Thus, the evaluation of both cultivars and breeding lines in the environmental trial showed that the pea breeding at the Plant Production Institute named after VYa Yuryev had a significant potential to create cultivars that would be well-adapted to both eastern and southern conditions, and that environmental trials remained an effective tool for assessing breeding material and selecting accessions with the maximum fulfillment of the genetic potential

Genetic Dissection of the Powdery Mildew Resistance in Wheat Breeding Line LS5082 Using BSR-Seq

Wheat powdery mildew is a destructive disease seriously threatening yield and quality. Comprehensive dissection of new resistance-related loci/genes is necessary to control this disease. LS5082 is a Chinese wheat breeding line with resistance to powdery mildew. Genetic analysis indicated that a single dominant gene, tentatively designated PmLS5082, conferred seedling resistance to different Bgt isolates. Bulked segregant RNA-seq (BSR-seq) was carried out to map the R gene PmLS5082 and profile differentially expressed genes associated with PmLS5082. PmLS5082 was mapped to chromosome arm 2BL and flanked by the markers WGGBH612-5 and YTU19-005 with genetic distances of 0.3 and 0.4 cM, respectively. The physical position was subsequently locked into the interval of 710.3-711.0 Mb. PmLS5082 differs from the catalogued Pm genes on chromosome arm 2BL in its resistant spectrum, physical position and origin, suggesting it is most likely a new Pm gene. Through COG and KEGG analysis, differentially expressed genes associated with PmLS5082 were profiled, and six genes in PmLS5082 interval were confirmed to be associated with PmLS5082 via RT-qPCR, using an additional set of wheat samples and time-course analysis post-inoculation with Bgt. In order to transfer PmLS082 to different genetic backgrounds using marker-assisted selection (MAS), closely linked markers, including Kompetitive Allele-Specific PCR (KASP) markers, were evaluated and nine markers were confirmed to be widely applicable for MAS in different genetic backgrounds.

Seedling morphology of different wheat genotypes at early stages under hydrocultural conditions

Consuming “sprouted seeds” is one of the most important factors of a healthy diet. An experiment was conducted in the University of Debrecen, Research Centre of Nyíregyháza (Hungary) in 2014 to analyse some morphological traits of four winter wheat genotypes (Triticum aestivum L.) and one spelt (T. spelta) variety. Our results showed that the spelt wheat variety “Franckenkorn” could maintain higher root length throughout the experimental period. On average, both “Perbetei” and “Franckenkorn” varieties could maintain higher root number compared to the other varieties. The extensive breeding line “1401 HK” had the highest shoot length throughout the whole experiment, being significantly higher than the landrace variety “Perbetei” and both of the varieties “KG Bendegúz” and “KG Kunhalom”. It could be concluded that “KG Bendegúz” cultivar and “Perbetei” landrace seem to be the most suitable for aquaculture techniques. In addition, “1401 HK” breeding line can be the most suitable for the production of juice since the minimal required shoot length (12 cm) was achieved within the shortest period of time after sowing (9 days). This breeding line and “Franckenkorn” can also be suitable for production of “wheatgrass”, because it is consumed without roots. Further research is needed to evaluate nutritional values of these genotypes.