RNAi-derived transgenic resistance to Mungbean yellow mosaic India virus in cowpea
Open Access
- 27 October 2017
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 12 (10), e0186786
- https://doi.org/10.1371/journal.pone.0186786
Abstract
Cowpea is an important grain legume crop of Africa, Latin America, and Southeast Asia. Leaf curl and golden mosaic diseases caused by Mungbean yellow mosaic India virus (MYMIV) have emerged as most devastating viral diseases of cowpea in Southeast Asia. In this study, we employed RNA interference (RNAi) strategy to control cowpea-infecting MYMIV. For this, we generated transgenic cowpea plants harbouring three different intron hairpin RNAi constructs, containing the AC2, AC4 and fusion of AC2 and AC4 (AC2+AC4) of seven cowpea-infecting begomoviruses. The T0 and T1 transgenic cowpea lines of all the three constructs accumulated transgene-specific siRNAs. Transgenic plants were further assayed up to T1 generations, for resistance to MYMIV using agro-infectious clones. Nearly 100% resistance against MYMIV infection was observed in transgenic lines, expressing AC2-hp and AC2+AC4-hp RNA, when compared with untransformed controls and plants transformed with empty vectors, which developed severe viral disease symptoms within 3 weeks. The AC4-hp RNA expressing lines displayed appearance of milder symptoms after 5 weeks of MYMIV-inoculation. Northern blots revealed a positive correlation between the level of transgene-specific siRNAs accumulation and virus resistance. The MYMIV-resistant transgenic lines accumulated nearly zero or very low titres of viral DNA. The transgenic cowpea plants had normal phenotype with no yield penalty in greenhouse conditions. This is the first demonstration of RNAi-derived resistance to MYMIV in cowpea.Funding Information
- Department of Biotechnology, Government of India (BT/173/NE/TDP/2011 and BT/PR13560/COE/34/44/2015)
This publication has 74 references indexed in Scilit:
- Mungbean yellow mosaic virus (MYMV) AC4 suppresses post-transcriptional gene silencing and an AC4 hairpin RNA gene reduces MYMV DNA accumulation in transgenic tobaccoVirus Genes, 2013
- Improved Agrobacterium-mediated transformation of cowpea via sonication and vacuum infiltrationPlant Cell Reports, 2011
- Geminivirus mixed infection on pepper plants: Synergistic interaction between PHYVV and PepGMVVirology Journal, 2011
- Engineering Resistance Against Mungbean yellow mosaic India virus Using Antisense RNAIndian Journal of Virology, 2010
- Roles of Plant Small RNAs in Biotic Stress ResponsesAnnual Review of Plant Biology, 2009
- RNA Silencing against Geminivirus: Complementary Action of Posttranscriptional Gene Silencing and Transcriptional Gene Silencing in Host RecoveryJournal of Virology, 2009
- Cowpea golden mosaic disease in Gujarat is caused by a Mungbean yellow mosaic India virus isolate with a DNA B variantArchiv für die gesamte Virusforschung, 2008
- Antiviral Immunity Directed by Small RNAsCell, 2007
- Tomato Chlorotic Mottle Virus Is a Target of RNA Silencing but the Presence of Specific Short Interfering RNAs Does Not Guarantee Resistance in Transgenic PlantsJournal of Virology, 2007
- Cowpea [ Vigna unguiculata (L.) Walp.]Published by Taylor & Francis Ltd ,2005