The wheat wild relative Aegilops tauschii was previously used to transfer the Lr42 leaf rust resistance gene into bread wheat. Lr42 confers resistance at both seedling and adult stages, and it is broadly effective against all leaf rust races tested to date. Lr42 has been used extensively in the CIMMYT international wheat breeding program with resulting cultivars deployed in several countries. Here, using a bulked segregant RNA-Seq (BSR-Seq) mapping strategy, we identify three candidate genes for Lr42. Overexpression of a nucleotide-binding site leucine-rich repeat (NLR) gene AET1Gv20040300 induces strong resistance to leaf rust in wheat and a mutation of the gene disrupted the resistance. The Lr42 resistance allele is rare in Ae. tauschii and likely arose from ectopic recombination. Cloning of Lr42 provides diagnostic markers and over 1000 CIMMYT wheat lines carrying Lr42 have been developed documenting its widespread use and impact in crop improvement.
Aegilops tauschii (2n = 2x = 14, genome DD), also known as Tausch’s goatgrass, is the D genome donor of bread or hexaploid wheat Triticum aestivum (2n = 2x = 42, AABBDD genome). It is a rich reservoir of useful genes for biotic and abiotic stress tolerance for wheat improvement. We developed a TILLING (Targeting Induced Local Lesions In Genomes) resource for Ae. tauschii for discovery and validation of useful genes in the D genome of wheat. The population, referred to as TILL-D, was developed with ethyl methanesulfonate (EMS) mutagen. The survival rate in M1 generation was 73%, out of which 22% plants were sterile. In the M2 generation 25% of the planted seeds showed phenotypic mutations such as albinos, chlorinas, no germination, variegated, sterile and partially fertile events, and 2,656 produced fertile M2 plants. The waxy gene was used to calculate the mutation frequency (1/70 kb) of the developed population, which was found to be higher than known mutation frequencies for diploid plants (1/89–1/1000 kb), but lower than that for a polyploid species (1/24–1/51 kb). The TILL-D resource, together with the newly published Ae. tauschii reference genome sequence, will facilitate gene discoveries and validations of agronomically important traits and their eventual fine transfer in bread wheat.
Soil alkalinity is an important stressor that impairs crop growth and development, resulting in reduced crop productivity. Unlike salinity stress, research efforts to understand the mechanism of plant adaptation to alkaline stress is limited in rice, a major staple food for the world population. We evaluated a population of 193 recombinant inbred lines (RIL) developed from a cross between Cocodrie and N22 under alkaline stress at the seedling stage. Using a linkage map consisting of 4849 SNP markers, 42 additive QTLs were identified. There were seven genomic regions where two or more QTLs for multiple traits colocalized. Three important QTL clusters were targeted, and several candidate genes were identified based on high impact variants using whole genome sequences (WGS) of both parents and differential expression in response to alkalinity stress. These genes included two expressed protein genes, the glucan endo-1,3-beta-glucosidase precursor, F-box domain-containing proteins, double-stranded RNA-binding motif-containing protein, aquaporin protein, receptor kinase-like protein, semialdehyde hydrogenase, and NAD-binding domain-containing protein genes. Tolerance to alkaline stress in Cocodrie was most likely due to the low Na+/K+ ratio resulting from reduced accumulation of Na+ ions and higher accumulation of K+ in roots and shoots. Our study demonstrated the utility of integrating QTL mapping with WGS to identify the candidate genes in the QTL regions. The QTLs and candidate genes originating from the tolerant parent Cocodrie should be targeted for introgression to improve alkalinity tolerance in rice and to elucidate the molecular basis of alkali tolerance.
Genetic improvement of crop plants is dependent on the availability of genetic variation, which can be present naturally or can be created using mutations. Ethyl methanesulfonate (EMS) mutagenesis provides an attractive tool to create variant alleles of genes of interest in any plant. In the present paper, we report development of an EMS‐mutagenized population in a hard red winter wheat (Triticum aestivum L.) variety Jagger. Jagger has been a leading variety across the United States and has been grown internationally as well, for over two decades now. It possesses many useful traits related to disease resistance, cold tolerance, and milling and baking qualities. Because it is a landmark variety, the genome of Jagger is being sequenced by an international collaborative team, which will speed up gene mapping and cloning efforts in it. The Jagger Targeting Induced Local Lesions In Genome (TILLING) resource developed in this work will be useful for functional validation of these genes. The mutagenized population was developed using EMS and was characterized phenotypically and genotypically. Gene‐wise mutation screening was performed for three genes located on different chromosomes across all the three genomes. The population has an overall mutation frequency of 1/40.7 kb, and we were able to find multiple knockout, missense, and missplicing mutations for each targeted gene. The population will be useful as a forward and reverse genetic screen for a number of valuable traits.
Use of genetic resistance is one of the most important strategies to manage the devastating disease Fusarium head blight (FHB) in wheat. Numerous quantitative trait loci (QTL) having varying effects on reducing FHB and the mycotoxin deoxynivalenol (DON) accumulation have been reported from Asian, European, or distant sources such as wild relatives of wheat (Triticum aestivum L.). However, coming from nonadapted backgrounds, the incorporation of such QTL into regional breeding programs has often resulted in the simultaneous transfer of other undesirable traits. Therefore, it is important to identify, characterize, and deploy sources of genetic resistance that do not suffer from poor adaptability and/or linkage drag. In the present work, QTL associated with FHB resistance in a high‐yielding, moderately resistant soft red winter wheat cultivar ‘Jamestown’ were mapped and validated. The QTL mapping was done using a recombinant inbred line (RIL) population of Pioneer ‘25R47’ × Jamestown having 186 individuals. Phenotyping over 2 yr at three locations, and genotyping using the 90K single nucleotide polymorphism (SNP) platform identified two new QTL, named QFHB.vt‐1B.1 and QFHB.vt‐1B.2, on the chromosome 1B long arm. The QTL contributed to FHB incidence, FHB severity, Fusarium‐damaged kernels, and DON content. Independent mapping of these QTL using two additional RIL populations of FG95195 × Jamestown (170 RILs) and Jamestown × LA97113UC‐124 (77 RILs) validated their stability and effectiveness in different genetic backgrounds. Kompetitive allele specific polymerase chain reaction (KASP) assays were developed using linked SNPs for marker‐assisted selection of the QTL. These QTL are being used in breeding programs to develop FHB‐resistant, high‐yielding varieties.
Genetic improvement of crop plants is dependent on the availability of genetic variation, which can be present naturally or can be created using mutations. Ethyl methanesulfonate (EMS) mutagenesis provides an attractive tool to create variant alleles of genes of interest in any plant. In the present paper, we report development of an EMS‐mutagenized population in a hard red winter wheat (Triticum aestivum L.) variety Jagger. Jagger has been a leading variety across the United States and has been grown internationally as well, for over two decades now. It possesses many useful traits related to disease resistance, cold tolerance, and milling and baking qualities. Because it is a landmark variety, the genome of Jagger is being sequenced by an international collaborative team, which will speed up gene mapping and cloning efforts in it. The Jagger Targeting Induced Local Lesions In Genome (TILLING) resource developed in this work will be useful for functional validation of these genes. The mutagenized population was developed using EMS and was characterized phenotypically and genotypically. Gene‐wise mutation screening was performed for three genes located on different chromosomes across all the three genomes. The population has an overall mutation frequency of 1/40.7 kb, and we were able to find multiple knockout, missense, and missplicing mutations for each targeted gene. The population will be useful as a forward and reverse genetic screen for a number of valuable traits.
SummaryField experiments during the wet seasons of 1981–1985 determined the response of soya bean to the application of P, K and Mo at four locations in the Nigerian savanna. Soya bean was grown at factorial combinations of four rates of P (0, 13·2, 26·4 and 396 kg P/ha) and two rates of K. (0 or 41·5 kg K/ha) in 1981, but in subsequent years two or three rates of Mo (0, 0·5 or 1·0 kg ammonium molybdate/ha) were included in a randomized complete block design with three replications at all locations.Significant yield increases due to the application of P were recorded in 10 out of 11 trials. Yield increases due to 13·2 or 26·4 kg P/ha were from 29 to 210% over the control. The effects of K and Mo application were not significant in 10 out of 11 trials and eight out of nine trials, respectively. Whereas P x Mo interaction effects were significant in three trials only, highest yields were recorded due to application of 26·4 or 39·6 kg P/ha in combination with 0·5 kg ammonium molybdate/ha.Application of P increased the percentage of P in soya-bean leaves significantly and a range of 0·275–0·330% P appears to be the critical nutrient range of P in soya-bean leaves at full bloom stage. Application of K had no effect on K concentration in leaves.
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