alexa Transfer And Expression Analysis Of RTBV-O-Ds2 Viral Gene In Indian Rice Varieties Against Rice Tungro Virus (RTV) Using RNAi Technology And Chromosomal Location Of Transgene | 23681
ISSN: 1948-5964

Journal of Antivirals & Antiretrovirals
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4th World Congress on Virology

Jayanta Tarafdar
Accepted Abstracts: J Antivir Antiretrovir
DOI: 10.4172/1948-5964.S1.022
Abstract
Rice, Oryza sativa L., is one of the most important cereal crops in the world and rice tungro disease (RTD) is the most limiting factor for rice cultivation especially in South and South-East Asian countries. Rice tungro is a composite disease caused by simultaneous infection of plant with two unrelated viruses viz. rice tungro bacilliform virus (RTBV), a plant pararetro double-stranded (ds) DNA virus in the family Caulimoviridae, under the genus tungrovirus and rice tungro spherical virus (RTSV), a positive sense single-stranded (ss) RNA virus in the Sequiviridae family. Tungro viruses are efficiently transmitted in a semi-persistent manner by green leafhoppers (Nephotetiix virescens). In the present study RNAi based transgene construct, carrying RTBV ORF IV in both sense and anti-sense orientation, from a transgenic Pusa Basmati 1 line (PB1/RTBV-O-Ds2) was backcrossed with two popular rice varieties viz. Sadabdi (IET 4786) and Khitis ( IET 4094). Functionally, the transgene construct targets homology-dependent selective degradation of RTBV transcripts employing RNAi mechanism. The backcross (BC) generations (BC3F4) were developed from O-Ds2 positive lines. The PCR-mediated detection of transgene indicated successful transfer of the RNAi into the rice cultivars through repeated backcrossing and a high frequency of amplification of fragments specific to ORF-IV (Ods-2 gene) and for hpt marker gene was detected in the successive backcrossed progenies of the rice varieties. The reverse-transcription PCR (RT-PCR) analysis of the backcrossed progenies for RTBV-O-Ds2 was also performed to check the activation of RNAi strategy in case of backcrossed lines. The expression of the transgene in both the varieties was analyzed by real-time PCR and attempt has been taken to identify the flanking sequence of the T-DNA and its characterization. Keywords: Rice Tungro Disease, Ods-2 gene, expression, rice progenies
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