Research in Plant Disease 2012;18(4):268-276.
Published online December 30, 2012.
Transient Expression of Homologous Hairpin RNA Interferes with Broad bean wilt virus 2 Infection in Nicotiana benthamiana
Ju-Yeon Yoon†, Ki Hyun Ryu†, Seung-Kook Choi1, Gug Sun Choi1, Soon Bae Kwon2, Jin Woo Park3 and Peter Palukaitis*
PVGB, Department of Horticulture and Landscape, College of Natural Science, Seoul Women’s University, Seoul 139-774, Korea
1Horticultural & Herbal Environmental Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Suwon 441-440, Korea
2Ganwon Agricultural Research and Extension Services, Chunchon 200-939, Korea
3Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Korea
Received: October 30, 2012   Revised: November 15, 2012   Accepted: November 26, 2012
Abstract
Broad bean wilt virus 2 (BBWV2), genus Fabavirus, subfamily Comovirinae, family Secoviridae, causes damage in many economically important horticultural and ornamental crops. Sequence alignments showed several conserved sequences in 5' non-coding regions (5' NCRs) of RNA 1 and RNA 2 in all BBWV2 strains characterized so far. Based on this observation, we generated a hpRNA construct (pIR-BBWV2) harboring an inverted repeat containing a 210 bp cDNA fragment homologous to 5' NCR portion of BBWV2 RNA 1 to investigate the silencing potential for its ability to interfere with a rapidly replicating BBWV2. Agrobacteriummediated transient expression of the IR-BBWV2 had a detrimental effect on BBWV2 infection, showing no distinct symptoms in non-inoculated leaves of the agroinfiltrated Nicotiana benthamiana plants. BBWV2 genomic RNAs were not detected by RT-PCR from tissues of both the inoculated leaves and upper leaves of the agroinfiltrated plants. Accumulation of virus-derived small interfering RNAs was detected in the inoculated leaf tissues of N. benthamiana plants elicited by transient expression of IR-BBWV2 indicating that RNA silencing is responsible for the resistance to BBWV2.
Key Words: Agroinfiltration, Broad bean wilt virus 2, Resistance, RT-PCR, Small-interfering RNA, Transient expression
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