Resistance gene identification in Vigna species infected with Begomovirus
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Date
2024
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Punjab Agricultural University
Abstract
The successful intra and inter-specific hybridization in Vigna revealed the dominant nature of yellow mosaic disease resistance with both monogenic and digenic gene action with dominant epistasis in Vigna mungo and digenic gene action with complementary epistasis in Vigna radiata. Further, differential expression of different classes of resistance genes in both compatible and incompatible Vigna – mungbean yellow mosaic India virus interactions was investigated. The qPCR primers were designed for amplifying plant resistant protein domain - specific regions (NB-ARC; SgS3; RdRp3) from genome of different Vigna species. Resistance protein domain specific sequences were shortlisted (with candidate genes, type species) and retrieved from NCBI database, followed by multiple sequence alignment through MEGA11 software on different Vigna species genome. A total of 37 qPCR primers with three domain - specific regions were designed using Primer3Web version 4.1.0 online software, were amplifying with specific amplicon size (90 to195 bp) and moreover without primer dimer formation. Most of the primers were amplified on genomic DNA of four resistant and two
susceptible genotypes of different Vigna speices genome. Total RNA was isolated and cDNA panel was prepared from mock and MYMIV inoculated leaf tissues of both resistant and susceptible genotypes of different Vigna species at 1, 5, 10, 15, 20, 25 and 30 days post inoculations. SYBR green chemistry was performed in RT-qPCR to study differential gene expression in both compatible and incompatible interactions. All the three resistant protein domains i.e. NB-ARC, SgS3 and RDR were found to be involved in YMD resistance in Vigna radiata and Vigna umbellata with absence of NB-ARC functional, only in Vigna mungo. On the other hand, expression of mungbean yellow mosaic India virus specific coat protein gene AV1 was higher in susceptible Vigna radiata genotype (SML1082) as compared to Vigna mungo and vice-versa in case of replication gene AC1. Low level of expression of both the AV1 and AC1 genes was also observed in resistant genotypes of Vigna with first experimental evidence of Vigna umbellata as host of mungbean yellow mosaic India virus.
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Kuppuraj J (2024). Resistance gene identification in Vigna species infected with Begomovirus (Unpublished Ph.D. Dissertation). Punjab Agricultural University, Ludhiana, Punjab, India.