2011 Fiscal Year Final Research Report

Development of a reverse-genetics system for Tomato spotted wilt virus

Research Project

Project/Area Number	23658043
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Plant pathology
Research Institution	National Institute of Agrobiological Sciences
Principal Investigator	ISHIKAWA Masayuki 独立行政法人農業生物資源研究所, 植物・微生物間相互作用研究ユニット, ユニット長 (70192482)
Project Period (FY)	2011
Keywords	感染・増殖 / 植物 / マイナス鎖RNAウイルス / 複製 / 転写 / 宿主因子
Research Abstract	Toward establishing an experimental system in which infection with Tomato spotted wilt virus(TSWV) can be initiated from its cDNA, we expressed TSWV nucleocapsid protein and RNA polymerase(the L protein) in Escherichia coli and Saccharomyces cerevisiae, respectively. We purified these proteins and mixed with in vitro-synthesized TSWV RNA in various conditions, however, we could not detect RNA synthesis. We also constructed plasmids to express TSWV-coded proteins and RNA in vivo, and introduced these plasmids into Nicotiana benthamiana leaves by the Agro-infiltration method, infection with TSWV did not occur. In the course of these trials, we obtained results that suggest that the translation elongation factor eEF1A plays important roles in mRNA transcription of TSWV.