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http://dx.doi.org/10.5423/RPD.2012.18.4.268

Transient Expression of Homologous Hairpin RNA Interferes with Broad bean wilt virus 2 Infection in Nicotiana benthamiana  

Yoon, Ju-Yeon (PVGB, Department of Horticulture and Landscape, College of Natural Science, Seoul Women's University)
Ryu, Ki Hyun (PVGB, Department of Horticulture and Landscape, College of Natural Science, Seoul Women's University)
Choi, Seung-Kook (Horticultural & Herbal Environmental Division, National Institute of Horticultural and Herbal Science, Rural Development Administration)
Choi, Gug Sun (Horticultural & Herbal Environmental Division, National Institute of Horticultural and Herbal Science, Rural Development Administration)
Kwon, Soon Bae (Ganwon Agricultural Research and Extension Services)
Park, Jin Woo (Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration)
Palukaitis, Peter (PVGB, Department of Horticulture and Landscape, College of Natural Science, Seoul Women's University)
Publication Information
Research in Plant Disease / v.18, no.4, 2012 , pp. 268-276 More about this Journal
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. Agrobacterium-mediated 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.
Keywords
Agroinfiltration; Broad bean wilt virus 2; Resistance; RT-PCR; Small-interfering RNA; Transient expression;
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