Bulked segregant analysis was undertaken to tag gene(s) controlling rust resistance using molecular markers in cowpea, to permit rapid selection of superior desirable rust resistant genotypes in the breeding program. For this purpose, the C-152, cultivated variety with high yielding, semi determinate plant type, good protein content and highly rust susceptible was crossed with genotype IC202778, the landrace from Himachal Pradesh, India having determinate, semi spreading and rust resistant characters. The parental genotypes were analyzed with 92 SSR markers for detection of polymorphism and only 13 markers showed polymorphism between the parents. Using each of these 13 SSR primers, we carried out bulked segregate analysis on F 2 plants representing two extremes of rust disease resistance and susceptible trait. Three SSR markers VuUGM02, VuUGM08 and VuUGM19 were found to be associated with rust resistance. This was further confirmed through selective genotyping. The co-segregation data on these molecular markers and rust resistance on F 2 plants were analysed by means of single-marker linear regression approach. Significant regression suggested linkage between VuUGM02 and rust resistance gene.
In plants, the biosynthesis of the phenylpropanoid, flavonoid and fatty acid pathway monomers, polymers and conjugated metabolites play a vital role in disease resistance. These are generally deposited to reinforce cell walls to contain the pathogen to the site of infection. Identification of sequence variants in genes that biosynthesise these resistance metabolites can explain the mechanisms of disease resistance. The resistant and susceptible genotypes inoculated with Phytophthora infestans were RNA sequenced to identify the single nucleotide polymorphisms (SNPs) and insertion/deletion (InDel) variations. The SNPs/InDels were annotated and classified into different categories based on their effect on gene functions. In the selected 25 biosynthetic genes overlapping 39 transcripts, a total of 52 SNPs/InDels were identified in the protein-coding (CDS) regions. These were categorised as deleterious based on prediction of their effects on protein structure and function. The SNPs/InDels data obtained in this study can be used in genome editing to enhance late blight resistance in Russet Burbank and other potato cultivars.
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