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The combination of several additive, partial resistance genes has been proposed as a preferred strategy to breed wheat cultivars with acceptable levels of durable resistance. The German winter wheat cv. Centrum has displayed high levels of adult plant stripe rust resistance (APR) in field environments for many years. One hundred and fifty one F2:7 RILs were developed from a cross between susceptible landrace Mingxian169 and Centrum to determine the inheritance of the APR resistance. The RILs and parents were evaluated for maximum disease severity (MDS) in the field during the 2015-2016 and 2016-2017 cropping seasons. Affymetrix 35K single nucleotide polymorphism (SNP) arrays were used to genotype the RILs and parents. In addition, the 660K SNP array was used to genotype bulked extreme pools and parents for saturation mapping. Four stable QTL were detected in all tested environments using inclusive composite interval mapping (ICIM); namely QYrCEN.nwafu-4AL, QYrCEN.nwafu-4BS, QYrCEN.nwafu-7BL, and QYrCEN.nwafu-7DS. QYrCEN.nwafu-4BS and QYrCEN.nwafu-7DS were contributed by MX169, QYrCEN.nwafu-4AL and QYr-CEN.nwafu-7BL were contributed by Centrum. QYrCEN.nwafu-7BL and QYrCEN.nwafu-4AL appear to represent new APR loci based on map comparisons. QYrCEN.nwafu-4BS contributed by MX169 also seems to represent a new locus. QYrCEN.nwafu-7DS is likely Yr18. Although MX169 was fully susceptible in our nurseries it is positive for the widely used marker csLV34. Reference lines carrying Yr18 are moderately resistant. Our hypothesis is that MX169 probably carries the inhibitor reported in Chinese landrace varieties by Wu et al. (2015, Plant Breeding 134: 634-640). SNP markers within these QTL were converted to KASP markers and validated in a subset of 120 diverse lines. These KASP markers should be useful for marker-assisted selection to improve stripe rust resistance in breeding programs.
Leaf rust disease, caused by the fungal pathogen Puccinia tritcina, is the most destructive foliar disease of wheat worldwide. Gene combination of Lr37/Yr17/Sr38 has been used in Georgia (GA) to prevent the loss from leaf rust; however, with the emergence of new virulent races, these genes have lost their effectiveness. 'AGS 2000' and 'Pioneer 26R61' are the most common soft red winter wheat (SRWW) cultivars in Southeastern US, and have been used as good sources of resistance to leaf and stripe rusts, and powdery mildew. To characterize the genetic basic of resistance of AGS 2000, a mapping population of 178 recombinant inbred lines (RIL) has been developed from a cross with Pioneer 26R61. This population was genotyped using a combination of SSR, DArT, and SNP markers, and a total of 2734 markers covering the entire genome were used for the construction of genetic map. Phenotypic evaluation of parents and RIL population was conducted at the seedling stage using a virulent GA leaf rust race. QTL mapping revealed a major QTL on chromosome 2BL, explaining about 20% of total phenotypic variation in AGS 2000. Additionally, a minor QTL was also detected on chromosome 5B. QTL on 2BL was identified as a novel gene, and can be used in marker-assisted selection for leaf rust resistance.
Leaf rust (LR) and stem rust (SR) are threats to global wheat production and new races frequently overcome resistance genes deployed in wheat cultivars. Identification of new sources of resistance is a major goal for many pre-breeding programs. The objective of this study was to investigate the genetic basis of resistance to LR and SR in a diverse South American wheat panel. Molecular markers for known resistance genes and GBS were used to dissect genetic components. The wheat panel of 122 lines was characterized under field conditions at La Estanzuela Research Station, Uruguay, for disease severity (DS) to LR (2014 and 2015) and SR (2015), and LTN (leaf tip necrosis). Final DS for LR ranged between 0 and 95%, with mean values of 40% (2014) and 46% (2015). For SR, final DS ranged between 0 and 50%, with a mean value of 5%. The frequencies of positive diagnostic resistance markers among accessions were 20.5% for Lr34/Sr57, 6.6% for Lr68, 3.3% for Sr2/Lr27, 23% for Sr31/Lr26, 20.5% for Sr24/Lr24, 9.4% for Sr25/Lr19, and 0% for Sr39/Lr35. Of all the LR/SR resistance genes, only the effect of Lr68 was significant when predicting LR DS. Seventeen lines were identified with combinations of two genes, but no combination conferred a significantly improved level of resistance. Preliminary GWAS analysis for LR response on a subset of 86 lines revealed several QTLs, with a major QTL explained by Lr68. Lines with good levels of resistance to LR and SR, high expression of LTN, and absence of markers for the studied resistance genes were identified, indicating that there are other genes involved in resistance. Future research involving the testing of additional molecular markers for other known resistance genes, and a deeper GWAS analysis, will provide further information about the resistance genes present in this wheat panel.
Common wheat Arula displays an acceptable level of adult plant resistance (APR) to stripe rust (YR), leaf rust (LR) and stem rust (SR) in Mexico, and to SR (Ug99 races) in Kenya. A recombinant inbred line (RIL) population developed from the cross of Arula with susceptible parent Apav was phenotyped under artificially created epidemics of the three rusts in 2014, 2015 and 2016 in Mexico and for SR during the off and main seasons of 2015 in Kenya. The RIL population and parents were genotyped using an iSelect 90K SNP array and 3 gene-linked markers (Sr2/Yr30-gwm533; Lr34/Yr18/Sr57-csLV34; Lr68-csGS), and a genetic map of 2,634 markers was constructed to locate the resistance loci. Two consistent QTL contributed by Arula were detected on chromosomes 3BS and 7DS, which corresponded to the previously known APR genes Sr2/Yr30 and Lr34/Yr18/Sr57, respectively. Sr2/Yr30 explained 1.1-14.7% and 41.0-61.5% of the phenotypic variation for YR and SR, respectively; whereas Lr34/Yr18/Sr57 accounted for 22.5-78.0%, 40.0-84.3% and 13.8-24.8% of the phenotypic variation for YR, LR and SR, respectively. Arula was also found to carry the positive allele for marker csGS closely linked to gene Lr68 on chromosome 7BL, although this gene was not detected using composite interval mapping. Our results show that RILs possessing both Sr2/Yr30 and Lr34/Yr18/Sr57 had significantly enhanced APR to all three rusts in field trials conducted in Mexico and Kenya. Strategic utilization of these two pleiotropic, multi-pathogen resistance genes with other minor genes is recommended to develop durable rust resistant wheat cultivars.
Diploid A genome wheat species harbor immense genetic variability which has been targeted and proven useful in wheat crop improvement. Further, the development and deployment of sequence based markers in wheat using survey sequences from next generation sequencing has opened avenues for comparative analysis, gene transfer and marker assisted selection (MAS) using high throughput cost effective genotyping techniques. Chromosome 2A of wheat is known to harbor several economically important genes. The present study aimed at in silico identification of genes corresponding to full length cDNAs and mining of SSRs and ISBPs from 2A draft sequence assembly of Chinese Spring for marker development. In totality, 1029 primer pairs (478 gene based, 501 SSRs and 50 ISBPs) were used to screen for polymorphism in diploid A genome species i.e., T. monococcum and T. boeoticum that identified 221 polymorphic loci. Out of these, 119 markers were mapped in T. monococcum X T. boeoticum RIL population. The enriched 2A genetic map constituted 161 mapped markers with final map length of 549.6 cM. Further, the utility of this enriched genetic map was demonstrated towards the fine mapping of adult plant resistance (APR) QTL, QYrtm.pau-2A against stripe rust. Using composite interval mapping, a QTL was detected between G45 and G54 markers explaining 19% of phenotypic variance. The primer sequences of the two genic markers were used to find the scaffold of 343 kb from IWGSC WGA V0.4 data. Thirty five simple sequence repeat markers were designed from the scaffold sequence which are being used for the fine mapping of QYrtm.pau-2A.
Durum wheat (Triticum turgidum subsp. durum) landraces are rapidly disappearing from the main wheat production areas in the Fertile Crescent. Such local landraces are most likely contain geographically specific, ectopically adapted alleles or gene complexes for their harsh environments. A panel of 156 durum wheat landraces and released varieties were assembled from historical collections deposited in national and international gene banks and from a recent active collection mission from selected areas across Jordan. The panel were evaluated under field conditions in two different locations for one growing season. Data for days to heading, plant height, peduncle length, number of spikes spike length, spike weight, grains number, grains weight, number of kernels per spike and thousand-kernel weight were recorded. Results indicate the existence of a wide variation between the tested genotypes for all tested agronomical traits. For heading date, the Jordanian landrace "JDu103" was the earliest under dry environment conditions. Regarding grains weight and spike weight, the Jordanian landrace "JDu105" produced the highest mean value under humid conditions. Another landrace "JDu46" produced the longest spikes and the highest TKW mean value, while the Jordanian landrace "JDu105" produced the heaviest spikes weight mean value, while "JDu100" produced the highest grains number. For molecular analysis, total genomic DNA was extracted from each genotype and then used for SNP genotyping using Illumina iSelect wheat 90k SNP chip. Structure analysis showed that the analyzed durum wheat panel can be divided into three genetically distinct subgroups. The GWAS analysis identified 93 significant markers-traits associations for multiple traits with two QTLs located at 7A and 7B, which seems important for TKW in durum wheat under dry environments. In conclusion, the Jordanian landraces used in this study showed wide genotypic and phenotypic variability, which can be considered by plant breeders for their future use in breeding programs.
Resistance is the most economically viable approach to curb the threat of rusts in wheat. The defeat of Sr31 and vulnerability of other resistance genes to the highly virulent Pgt race Ug99 and variants led to renewed efforts to discover and deploy resistance genes/QTLs in new durably resistant varieties. Akuri is a CIMMYT-developed bread wheat line exhibiting adult plant resistance (APR) in field trials in Kenya despite susceptibility to many races at the seedling stage. This study was designed to identify genomic regions contributing APR to stem rust in Akuri. One hundred and forty one RILs and parents of an F2:5 Akuri x PBW343 population were evaluated in Njoro for APR to stem rust over three seasons. Composite interval mapping was implemented on Windows QTL Cartographer to detect QTLs at a LOD threshold of 2.5 utilizing 910 high quality SNPs previously typed on the DArT-GBS platform. Preliminary QTL analyses revealed loci on chromosomes 1B, 2B and 3B consistently contributing to stem rust resistance. These QTL respectively explained ~7, 9, and 8% of the phenotypic variation. A comparison with the recently reported QTL consensus map revealed that the QTL herein discovered are probably novel. Work is underway to saturate the identified genomic regions with microsatellite markers to identify candidate, linked markers for use in marker assisted selection (MAS)
The durability of stem rust resistance in wheat varieties is strengthened by the use of polygenic, and broad-spectrum sources of resistance. Adult plant resistance (APR) was observed in the mid-20th century Ecuadorian bread wheat cv. Morocho Blanco (PI 286545) in field tests at Njoro, Kenya, and at St. Paul. Morocho Blanco was susceptible to races TTKSK, RCRSC and TPMKC at the seedling stage. A doubled haploid (DH) mapping population was created from a cross between Morocho Blanco and the susceptible line LMPG-6 to identify loci associated with APR phenotypes. Eighty-eight DH lines were genotyped with approximately 90,000 SNPs using a custom Infinium assay from Illumina. Sixty-seven additional DH lines were used to verify SNPs associated with reduced stem rust levels. Severity and infection type were assessed on adult plants at the stem rust screening facility in Kenya in 2013 and 2014, and in two single race nurseries inoculated with races RCRSC and TPMKC at St. Paul in 2014. Two identified and verified QTL reducing stem rust severity were located on chromosome arms 2BS and 6AS. The QTL on 6AS also reduced infection type at Njoro, but a similar reduction was not observed at St. Paul suggesting a genotype x environment or genotype x race interaction. The QTL on 2BS was associated with reduced stem rust severity at both Njoro and St. Paul. It is a strong candidate for use in breeding for APR to stem rust.