ウメ輪紋ウイルス日本分離株の感染性cDNAクローンの構築および感染性cDNAクローンを用いた研究手法の確立

Plum pox virus (PPV) is regarded as an agriculturally important virus outside of Japan, and molecular biological analysis using infectious cDNA clones is widely performed for its pathogenicity analysis. Therefore, the purpose of this study is to construct infectious cDNA clones of PPV isolated in Japan and to establish research methods using infectious cDNA clones in Japanese apricot and Nicotiana plants. The cDNA clones of Japanese isolates using the 35S promoter were constructed, and the infectivity was confirmed in Nicotiana benthamiana. Then using the infectious cDNA clones, the research methods such as inoculating protoplasts of Japanese apricot and the leaves with particle delivery system (PDS) were established. Inoculation into protoplasts showed PPV infection and proliferation at the single-cell level, and inoculation with PDS showed cell-to-cell movement of PPV in the leaf. The chimeric clones between PPV Japanese isolates were constructed, and the inoculation test into N. benthamiana suggested that the 3´-latter half region of the PPV genome was involved in systemic infection.