2004-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/691936摘要：在台灣許多重要病害諸如花生簇葉病、甘藷簇葉病、梨樹衰弱病、水稻黃萎病等均為植物菌質體 (phytoplasmas) 所引起。長久以來，因植物菌質體均無法成功地被培養出來，且其細胞的純化仍有許多困難，因此有關其基本生物及生理、生化特性之研究仍屬有限，而有關植物菌質體與寄主植物交互作用分子生物學之探討更是付之闕如。在本計畫中將利用differential display及其他方法如subtractive hybridization及cDNA-AFLP等，以期能找出日日春感染花生簇葉病菌質體後基因表現差異的mRNA，並加以選殖cDNA，製作探針對基因庫進行篩選、進行南方氏雜配及北方雜配分析、核酸解序與比對等工作，深入探討該等基因之功能，並綜合評析日日春與花生簇葉病菌質體之間交互作用之關係。本計畫將於三年完成，分年之工作項目則包括： 第一年：完成花生簇葉病菌質體DNA之純化、基因庫之構築、模式植物日日春cDNA基因庫之構築。利用differential display方式篩選日日春受到花生簇葉病菌質體感染後，表現有差異的mRNA，並完成cDNA之選殖、製作探針對構築完成之基因庫進行篩選、南方<br> Abstract: In Taiwan, many important plant diseases such as peanut witches' broom (PNWB), sweetpotato witches' broom, pear decline, rice yellow dwarf were caused by phytoplasmas, former mycoplasmalike organisms (MLOs). Up to now, the phytopathogenic phytoplasmas still resist to be cultured in any available media. It's also very difficult to purify phytoplasmas from affected plants without the contamination of plant antigens. The biological and biochemical data of phytoplasmas are also next to nothing. In this study, differential display strategies will be applied to isolate periwinkle cDNAs differentially expressed following infection with PNWB-phytoplasma. Differentially expressed cDNAs will be cloned and characterized by Southern blot, Northern blot and sequence analysis. Gene functions and interactions between periwinkle and PNWB- phytoplasma will be further revealed. The proposed study will be accomplished in three years. In the first year, the genomic library of PNWB-phytoplasma and cDNA library of periwinkel will be constructed in lambda vector. Differential display strategies will be applied to isolate periwinkle cDNAs differentially expressed following infection with PNWB-phytoplasma. Differentially expressed cDNAs will be cloned and characterized by Southern blot, Northern blot and sequence analysis. In the second year, differentially expressed mRNAs will be screened by differential display and other approaches such as subtractive hybridization and cDNA-AFLP. In the third year, screening and analyzing of differentially expressed genes will be continued. The roles of these genes involved in the interaction between periwinkle and PNWB- phytoplama will be studied.植物菌質體日日春交互作用phytoplasmaperiwinkleinteraction