47.糸状菌のキメラジテルペン合成酵素の植物体内発現(口頭発表)

概要

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Fusicoccins are produced by plant-pathogenic fungus Phomopsis amygdali, and possess phytohormone-related activities. Recently, we identified an unusual chimeric diterpene synthase gene, PaFS, which is responsible for fusicoccin biosynthesis. PaFS encodes fusicoccadiene (biosynthesis intermediate cyclic hydrocarbon of fusicoccin) synthase (719 amino acids), possessing the geranylgeranyl diphosphate (GGDP) cyclase domain at the N-terminus and GGDP synthase (GGS) domain at the C-terminus. In this study, we tried to generate transgenic Arabidopsis thaliana plants expressing PaFS or N390 (N-terminus 390 amino acids of PaFS), possessing only diterpene cyclase activity, in plastids, containing abundant GGDP, or in the cytoplasm to examine wthether the fungal diterpenes are produced efficiently by the chimeric enzyme in plant cells. To target the fungal enzymes to plastids, a transit peptide (TP) of Arabidopsis copalyl diphosphate synthase was fused to their N-terminus. The four cDNA (PaFS, TP-PaFS, N390, TP-N390) were introduced to Arabidopsis through Agrobacterium infection. It was confirmed that each translated product from the transgene had proper activity by in vitro assay using bacterial recombinant protein, and RTPCR anylysis showed that each transcript was detected in T1 plants. We will further characterize each transgenic Arabidopsis using T2 plants.
2010-10-01