MOLECULAR CHARACTERIZATION OF BEGOMOVIRUS INFECTING SOYBEAN (GLYCINE MAX L.) CROP IN CENTRAL INDIA

Author(s): Brijesh K. Yadav, Sangeeta, Ritesh K. Jaiswal, Sunita Patel, and Achuit K. Singh

Abstract: Aim: To detect bipartite begomovirus infection in soybean crop, characterize the pathogen and demonstrate Koch`s postulate on Nicotiana benthamiana. Methods: Symptomatic soybean leaves were collected from fields; total DNA was isolated followed by PCR detection of begomovirus by degenerate primers. RCA product of viral genome was digested by restriction enzyme, cloned in pBluescript (pKS+) cloning vector and sequenced. Evolutionary study of begomo virus was analyzed by constructing phylogenetic tree. Partial tandem repeats were constructed in pCAMBIA2300 plant binary vector for demonstration of koch`s postulate. Results: Soybean crop was infected by bipartite begomo virus along with associated betasatellite. MYMIV DNA-A (2736nt) showed maximum (99.8%) nucleotide sequence identity with Mungbean yellow mosaic India virus (MYMIV-[IN-Ak-14] KP779635). MYMIV DNA-B (2671nt) showed maximum (99.8%) nucleotide sequence identity with Mungbean yellow mosaic India virus (MYMIV-[IN-Sat-12] KP779634). MYMIV DNA-β (1366nt) showed maximum (97.3%) nucleotide sequence identity with Tomato leaf curl Gandhinagar betasatellite (ToLCGNB-[IN-GN-12] KC952006). Evolutionary studies of DNA-A, DNA-B and DNA-β showed close relationship with previously reported bipartite begomoviruses. Upon agro-inoculation of partial tandem repeats of DNA-A, DNA-B and DNA-β, typical symptoms of begomovirus infection were observed on N. benthamiana and verified by RT-PCR in terms of accumulation of viral copy number. Conclusion: Soybean crop was found to be infected by bipartite begomovirus along with associated betasatellite of Tomato leaf curl Gandhinagar betasatellite. Agro-inoculation of partial tandem repeats of DNA-A, DNA-B and DNA-β was capable to produce and increase the symptom severity on N. benthamiana. This study will help to develop a resistant variety of soybean cultivar to enhance soybean crop yield.

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