Identification of Wheat Septoria tritici Resistance Genes in Wheat Germplasm Using Molecular Markers.

Zymoseptoria tritici (Z. tritici) is the main threat to global food security; it is a fungal disease that presents one of the most serious threats to wheat crops, causing severe yield losses worldwide, including in Kazakhstan. The pathogen leads to crop losses reaching from 15 to 50%. The objectives of this study were to (1) evaluate a wheat collection for Z. tritici resistance during the adult plant and seedling growth stages, (2) identify the sources of resistance genes that provide resistance to Z. tritici using molecular markers linked to Stb genes, and (3) identify potentially useful resistant wheat genotypes among cultivars and advanced breeding lines. This study evaluated 60 winter and spring wheat genotypes for Z. tritici resistance. According to the field reactions, 22 entries (35.7%) showed ≤10% disease severity in both years. The resistant reaction to a mix of Z. tritici isolates in the seedling stage was associated with adult plant resistance to disease in four wheat entries. The resistance of Rosinka 3 was due to the presence of Stb8; Omskaya 18 showed an immune reaction in the field and a moderately susceptible reaction in the seedling stage, possibly provided by a combination of the Stb7 and Stb2 genes. The high resistance in both the adult and seedling stages of Omskaya 29 and KR11-03 was due to the Stb4 and Stb2 genes and, possibly, due to the presence of unknown genes. A linked marker analysis revealed the presence of several Stb genes. The proportion of wheat entries with Stb genes was quite high at twenty-seven of the genotypes tested (45.0%), including four from Kazakhstan, nine from Russia, nine from the CIMMYT-ICARDA-IWWIP program, and five from the CIMMYT-SEPTMON nursery. Among the sixty entries, ten (16.7%) carried the resistance genes Stb2 and Stb8, and the gene Stb4 was found in seven cultivars (11.6%). Marker-assisted selection can be efficiently applied to develop wheat cultivars with effective Stb gene combinations that would directly assist in developing durable resistance in Kazakhstan. Resistant genotypes could also be used as improved parents in crossing programs to develop new wheat cultivars.

QTL mapping for seedling and adult plant resistance to stripe and leaf rust in two winter wheat populations.

The two recombinant inbred line (RIL) populations developed by crossing Almaly × Avocet S (206 RILs) and Almaly × Anza (162 RILs) were used to detect the novel genomic regions associated with adult plant resistance (APR) and seedling or all-stage resistance (ASR) to yellow rust (YR) and leaf rust (LR). The quantitative trait loci (QTLs) were detected through multi-year phenotypic evaluations (2018-2020) and using high-throughput DArTseq genotyping technology. RILs exhibited significant genetic variation with p < 0.001, and the coefficient of variation ranged from 9.79% to 47.99% for both LR and YR in all Environments and stages of evaluations. The heritability is quite high and ranged between 0.47 and 0.98. We identified nine stable QTLs for YR APR on chromosomes 1B, 2A, 2B, 3D, and 4D and four stable QTLs for LR APR on chromosomes 2B, 3B, 4A, and 5A. Furthermore, in silico analysis revealed that the key putative candidate genes such as cytochrome P450, protein kinase-like domain superfamily, zinc-binding ribosomal protein, SANT/Myb domain, WRKY transcription factor, nucleotide sugar transporter, and NAC domain superfamily were in the QTL regions and probably involved in the regulation of host response toward pathogen infection. The stable QTLs identified in this study are useful for developing rust-resistant varieties through marker-assisted selection (MAS).

Phenotyping and Identification of Molecular Markers Associated with Leaf Rust Resistance in the Wheat Germplasm from Kazakhstan, CIMMYT and ICARDA.

Leaf rust (LR) is the most widespread disease of common wheat worldwide. In order to evaluate leaf rust resistance, 70 uncharacterized wheat cultivars and promising lines with unknown leaf rust resistance genes (Lr genes) were exposed to Kazakhstani Puccinia triticina (Pt) races at the seedling stage. Field tests were performed to characterize leaf rust responses at the adult plant growth stage in the 2020-2021 and 2021-2022 cropping seasons. The wheat collection showed phenotypic diversity when tested with two virulent races of Pt. Thirteen wheat genotypes (18.6%) showed high resistance at both seedling and adult plant stages. In most cases, breeding material originating from international nurseries showed higher resistance to LR. Nine Lr genes, viz. Lr9, Lr10, Lr19, Lr26, Lr28, Lr34, Lr37, Lr46, and Lr68, either singly or in combination, were identified in 47 genotypes. Known Lr genes were not detected in the remaining 23 genotypes. The most commonly identified resistance genes were Lr37 (17 cultivars), Lr34 (16 cultivars), and Lr46 (10 cultivars), while Lr19, Lr68, Lr26, and Lr28 were the least frequent. Four Lr genes were identified in Keremet and Hisorok, followed by three Lr genes in Aliya, Rasad, Reke, Mataj, Egana and Almaly/Obri. The molecular screening revealed twenty-nine carriers of a single Lr gene, ten carriers of two genes, six carriers of three genes, and two carriers of four genes. Most of these accessions showed a high and moderate level of APR (Adult plant resistance) and may be utilized for the incorporation of Lr genes in well-adapted wheat cultivars. The most effective combination was Lr37, Lr34, and Lr68, the carriers of which were characterized by a low disease susceptibility index. The obtained results will facilitate breeding programs for wheat resistance in Kazakhstan.

Identification of Stripe Rust Resistance Genes in Common Wheat Cultivars and Breeding Lines from Kazakhstan.

Stripe (yellow) rust, caused by Puccinia striiformis f.sp. tritici (Pst), is a fungal disease that presents one of the most serious threats to the wheat crops, causing severe yield losses worldwide, including Kazakhstan. The objectives of this study were to: (1) evaluate a winter wheat collection for stripe rust resistance during an adult plant growth stage, (2) identify the presence of selected Yr genes using linked molecular markers in wheat germplasm, (3) identify potentially useful resistant wheat genotypes among leading cultivars and advanced breeding lines. This study evaluated 70 winter wheat genotypes for stripe rust resistance. According to the field reactions, 42 entries (60%) had R or MR reactions including 27 breeding lines (38.6%) and 15 (21.4%) cultivars. Twenty-eight breeding lines/cultivars (40.0%) were susceptible in both years. According to the average coefficient of infection value (ACI) six genotypes were regarded as possessing high level of adult plant resistance. Cultivars/lines carrying Yr10 alone or in combination with other Yr resistance genes provided resistance to stripe rust. Eleven breeding lines showed <5% disease severity in both years. Linked marker analysis revealed the presence of several gene and gene complexes (Yr5, Yr10, Yr15, Yr17/Lr37/Sr38 and Yr18/Lr34). Among a collection of 70 winter wheat breeding lines and cultivars produced in Kazakhstan three stripe rust resistance genes (Yr10, Yr5 and Yr15) demonstrated high frequency occurrence (31.4%, 14.0% and 7.0%, respectively). The most abundant was gene Yr10 identified in 22 genotypes. It was followed by the Yr5 gene, which conferred resistance in 14 lines (20%) and Yr18 gene-11 lines (15.7%). Yr15 was identified in 7 genotypes. Yr17/Lr37/Sr38 gene complex was found in 2 entries. Among 70 evaluated germplasm sources, 42 disease resistant entries are potentially useful resistant wheat genotypes. These carriers of different Yr genes can be used directly in breeding programs to improve stripe rust resistance of winter wheat. Marker-assisted selection can be efficiently applied to develop wheat cultivars with effective gene combinations that would directly assist in developing durable resistance in Kazakhstan.

Evaluation of Wheat Germplasm for Resistance to Leaf Rust (Puccinia triticina) and Identification of the Sources of Lr Resistance Genes Using Molecular Markers.

Leaf rust, caused by Puccinia triticina (Ptr), is a significant disease of spring wheat spread in Kazakhstan. The development of resistant cultivars importantly requires the effective use of leaf rust resistance genes. This study aims to: (i) determine variation in Ptr population using races from the East Kazakhstan, Akmola, and Almaty regions of Kazakhstan; (ii) examine resistance during seedling and adult plant stages; and (iii) identify the sources of Lr resistance genes among the spring wheat collection using molecular markers. Analysis of a mixed population of Ptr identified 25 distinct pathotypes. Analysis of these pathotypes using 16 Thatcher lines that are near-isogenic for leaf rust resistance genes (Lr) showed different virulence patterns, ranging from least virulent "CJF/B" and "JCL/G" to highly virulent "TKT/Q". Most of the pathotypes were avirulent to Lr9, Lr19, Lr24, and Lr25 and virulent to Lr1, Lr2a, Lr3ka, Lr11, and Lr30. The Ptr population in Kazakhstan is diverse, as indicated by the range of virulence observed in five different races analyzed in this study. The number of genotypes showed high levels of seedling resistance to each of the five Ptr races, thus confirming genotypic diversity. Two genotypes, Stepnaya 62 and Omskaya 37, were highly resistant to almost all five tested Ptr pathotypes. Stepnaya 62, Omskaya 37, Avangard, Kazakhstanskaya rannespelaya, and Kazakhstanskaya 25 were identified as the most stable genotypes for seedling resistance. However, most of the varieties from Kazakhstan were susceptible in the seedling stage. Molecular screening of these genotypes showed contrasting differences in the genes frequencies. Among the 30 entries, 22 carried leaf rust resistance gene Lr1, and two had Lr9 and Lr68. Lr10 and Lr28 were found in three and four cultivars, respectively. Lr19 was detected in Omskaya 37. Two single cultivars separately carried Lr26 and Lr34, while Lr37 was not detected in any genotypes within this study. Field evaluation demonstrated that the most frequent Lr1 gene is ineffective. Kazakhstanskaya 19 and Omskaya 37 had the highest number of resistance genes: three and four Lr genes, respectively. Two gene combinations (Lr1, Lr68) were detected in Erythrospermum 35 and Astana. The result obtained may assist breeders in incorporating effective Lr genes into new cultivars and developing cultivars resistant to leaf rust.