Next‐generation sequencing for identification of candidate genes for Fusarium wilt and sterility mosaic disease in pigeonpea (Cajanus cajan) |
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Authors: | Vikas K Singh Aamir W Khan Rachit K Saxena Vinay Kumar Sandip M Kale Pallavi Sinha Annapurna Chitikineni Lekha T Pazhamala Vanika Garg Mamta Sharma Chanda Venkata Sameer Kumar Swathi Parupalli Suryanarayana Vechalapu Suyash Patil Sonnappa Muniswamy Anuradha Ghanta Kalinati Narasimhan Yamini Rajeev K Varshney |
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Institution: | 1. International Crops Research Institute for the Semi‐Arid Tropics (ICRISAT), Patancheru, India;2. Agricultural Research Station (ARS)‐Gulbarga, University of Agricultural Sciences (UAS), Raichur, Karnataka, India;3. Institute of Biotechnology, Professor Jayshankar Telangana State Agricultural University (PJTSAU), Telangana, India;4. School of Plant Biology and Institute of Agriculture, The University of Western Australia, Crawley, WA, Australia |
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Abstract: | 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 × 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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Keywords: | Fusarium wilt sterility mosaic disease bulked segregant analysis whole‐genome re‐sequencing SNP index nonsynonymous SNPs |
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