IDENTIFICATION AND CLASSIFICATION OF VARIOUS OYVMV (OKRA YELLOW VEIN MOSAIC VIRUS) ISOLATES FROM DIFFERENT AGROCLIMATIC ZONES OF GUJARAT USING NEXT GENERATION SEQUENCING
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Date
2019
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DEPARTMENT OF AGRICULTURAL BIOTECHNOLOGY B. A. COLLEGE OF AGRICULTURE ANAND AGRICULTURAL UNIVERSITY ANAND
Abstract
The present investigation entitled “Identification and classification of various
OYVMV (Okra Yellow Vein Mosaic Virus) isolates from different agro-climatic
zones of Gujarat using Next Generation Sequencing” was carried out at the
Department of Agricultural Biotechnology, Anand Agricultural University, Anand
during 2017-19. During June-November 2018, total sixteen samples of OYVMV
infected okra leaves were collected from eight different agro-climatic zones of Gujarat
having okra under cultivation.
Viral DNA isolation from Yellow Vein Mosaic Virus infected okra leaf samples
was carried out by Extract-n-Amp Plant PCR kit. After isolation of viral genomic DNA
from sixteen samples, gel electrophoresis was carried out using 0.8% agarose gel. The
DNA bands corresponding to viral genome from the 16 different viral DNA samples
run on the agarose gel were eluted and purification through promega purification kit.
the ssDNA was quantified by NanoDrop® ND-1000. The Navsari samples gave the
highest quantity of viral genomic ssDNA (44.7 ng/μl) followed by Mehsana (43.3
ng/μl). The ssDNA was converted into dsDNA by using cDNA synthesis kit. After
purification, the Qubit® 3.0 Fluorometer was used to quantify the viral genomic dsDNA
and the highest quantity was recorded by sample from Kheda (38.3 ng/μl) followed by
Navsari (29.2 ng/μl).