Next-generation sequencing for identification of candidate genes for Fusarium wilt and sterility mosaic disease in pigeonpea (Cajanus cajan)
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Vikas Singh, Aamir Khan, Rachit Saxena, Vinay Kumar, Sandip Kale, Sinha Pallavi, Annapurna Chitikineni, Pazhamala Lekha, Garg Vanika, Chanda Venkata Sameer Kumar, Swathi Parupalli, Suryanarayana Vechalapu, Suyash Patil, Sonnappa Muniswamy, Anuradha Ghanta, Kalinati Narasimhan Yamini, Pallavi Subanna Dharmaraj, Rajeev Varshney. (31/5/2016). Next-generation sequencing for identification of candidate genes for Fusarium wilt and sterility mosaic disease in pigeonpea (Cajanus cajan). Plant Biotechnology Journal, 14(5), pp. 1183-1194.
To map resistance genes for Fusarium wilt (FW) and sterility mosaic disease (SMD) in pigeonpea, sequencing-based bulked segregant analysis (Seq-BSA) was used. Resistant (R) and susceptible (S) bulks from the extreme recombinant inbred lines of ICPL 20096 9 ICPL 332 were sequenced. Subsequently, SNP index was calculated between R- and S-bulks with the help of draft genome sequence and reference-guided assembly of ICPL 20096 (resistant parent). Seq-BSA has provided seven candidate SNPs for FW and SMD resistance in pigeonpea. In parallel, four additional genotypes were re-sequenced and their combined analysis with R- and S-bulks has provided a total of 8362 nonsynonymous (ns) SNPs. Of 8362 nsSNPs, 60 were found within the 2-Mb ﬂanking regions of seven candidate SNPs identiﬁed through Seq-BSA. Haplotype analysis narrowed down to eight nsSNPs in seven genes. These eight nsSNPs were further validated by re-sequencing 11 genotypes that are resistant and susceptible to FW and SMD. This analysis revealed association of four candidate nsSNPs in four genes with FW resistance and four candidate nsSNPs in three genes with SMD resistance. Further, In silico protein analysis and expression proﬁling identiﬁed two most promising candidate genes namely C.cajan_01839 for SMD resistance and C.cajan_03203 for FW resistance. Identiﬁed candidate genomic regions/ SNPs will be useful for genomics-assisted breeding in pigeonpea.
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