National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
PCR was employed to establish association of begomovirus through amplification of geminivirus specific PCR product. In order to determine the complete nucleotide sequence of DNA-A component of HgYMV, several universal primers/ abutting primers and specific primers available in the literature were tried to amplify full length DNA-A of 2.7 kb. The amplification of full length of DNA-A component of HgYMV was achieved with all the primers such as AC-abut and AV-abut, HgYMVAF and HgYMVAR and HYMV-A1500F and HYMV-A1500R. Virus specific DNA fragments of approximately 2700 bp were obtained from DNA of infected horsegram samples. No PCR product was obtained from DNA extracted from healthy samples and water control. An annealing temperature of 55º C for 2 min was found suitable for amplification of full length of DNA-A component of HgYMV. The PCR product (approx. 2700 bp) of DNA-A component of HgYMV amplified with HgYMVAF and HgYMVAR primers was cloned, sequenced and assembled and its length was determined as 2654 nucleotides. The nucleotide sequences of DNA-A component of horsegram yellow mosaic virus were compared with those of selected begomoviruses obtained from the NCBI database. Comparison of the complete DNA-A sequence of HgYMV-GKVK-Bangalore with other bipartite begomoviruses revealed 82-84 per cent identity with isolates of Mungbean yellow mosaic virus (MYMV), 78-81 per cent with Mungbean yellow mosaic India virus (MYMIV), 98 and 95 per cent identity with the accessions of horsegram and frenchbean isolates of HgYMV from Tamil Nadu and Srilanka, respectively. The phylogenetic tree built using the DNA-A of HgYMV-GKVK-Bangalore showed three clusters, with HgYMV-GKVK-Bangalore falling in cluster II, Mungbean yellow mosaic virus (MYMV) isolates in cluster I and Mungbean yellow mosaic India virus (MYMIV isolates) in cluster III. Analyses of HgYMV-DNA-A-GKVK-Bangalore isolate sequence showed typical features of bipartite begomoviruses characterized by six conserved open reading frames in DNA-A.
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