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標題: 台灣地區水稻紋枯病菌菌株差異性之研究
Studies on the diversity of Rhizoctonia solani AG1-IA isolates from Taiwan
作者: 楊相哲
Yang, Siang-Jhe
關鍵字: Rhizoctonia solani
出版社: 植物病理學系所
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摘要: 摘要 水稻病害是影響水稻產量的重要原因,其中,稻紋枯病為台灣稻作主要病害之一。水稻紋枯病由立枯絲核菌(Rhizoctonia solani AG 1-IA)所引起,而立枯絲核菌(Rhizoctonia solani Kühn)為絲核菌屬中一重要之集合種,目前已區分為14個菌絲融合群,分別為AG-1到AG-13及AG-BI。紋枯病之病害發展在初期以初次感染源菌核的密度影響發病,而二次感染源為菌絲。本試驗在收集來自台灣各地區水稻紋枯病菌後,培養在不同溫度(8、12、16、20、24、28、32、36 ℃)、不同酸鹼值(pH 4、5、6、7、8、9)的情形下,觀察紀錄其病原菌之菌絲生長與菌核數量、菌核乾重,了解其生理特性。結果發現菌絲最適生長溫度為28℃,其菌落大小平均為67.3625 mm/day,菌絲平均每小時生長速率為1.198 mm。在菌絲融合群(MCGs)試驗方面,依照菌絲間的親和性將台灣水稻紋枯病菌分為四群Group I ~ IV,另外,利用單一引子聚合酵素連鎖反應(Single primer PCR)增幅出差異性片段,應用電腦軟體依UPGMA分群法對各菌株進行差異性比較,發現相似度在相似度68.8%時,就能夠區分出台灣水稻紋枯病菌AG1-IA與其它融合群,而再深入觀察,在相似度86%時可將台灣水稻紋枯病菌大致分成五群:第一群為N群 (Group north),主要採集自北部地區(包括宜蘭縣、桃園縣及新竹縣市)的菌株;第二群為M群 (Group midland),主要採集自中南部(包括台中縣市、南投縣、彰化縣、雲林縣及嘉義縣市)的菌株;第三群為E群 (Group east),主要採集自東部地區(包括花蓮縣及台東縣)的菌株;第四群為S群 (Group south),主要採集自南部地區(包括嘉義縣市、台南縣市及屏東縣)的菌株;第五群為C群 (Group complex),為較複雜的一群,地理位置不一。由MCGs及SSR-PCR的分群,發現台灣水稻紋枯病菌菌株間差異性主要依地理環境而不同,且環境上的地理區隔對菌株的遺傳差異性影響遠比寄主(水稻品種)因素大。未來可持續收集台灣各地區菌株,建立紋枯病菌菌株資料庫,作為水稻防治及育種之用。
ABSTRACT Rice disease is the major reason for decreasing the harvest of rice, and the rice sheath blight is an important disease of rice disease in Taiwan. The pathogen is Rhizoctonia solani AG1-IA, and Rhizoctonia solani is a large species complex. At lest, there are 14 anastomosis groups (AGs) including AG-1 through AG-13 and AG-BI (bridging isolate). Sclerotia is the first infection source of the initial stage of rice disease, and mycelium is the second infection source of ice disease. The purposes of the thesis were to collect isolates of Rhizoctonia solani AG1-IA from Taiwan and to botanize these isolates about the diversity among isolates. About physiological characteristic test, Isolates were cultured on PDA mediums at various temperature (8, 12, 16, 20, 24, 28, 32, 36 ℃) and pH value (pH 4, 5, 6, 7, 8, 9), to botanize mycelial grouth rate and dry weight of sclerotia, and numbers of sclerotia. The result suggested that the mycelial growth of all isolates was found best at 28 ℃, and the average colony size was 67.3625 mm/day, and then the average mycelial growth rato rate was 1.198 mm/hr. About mycelial compatiable groups (MCGs) test, there were four groups in Taiwan by mycelial compatiable and incompatiable reation. On the other hand, using single primer PCR to amplify difference fragments and to analysis genetic similarity by by UPGMA cluster analysis with computer software Gel Compar. Dendrogram based on the analysis of 54 isolates of Rhizoctonia solani AG1-IA, Lentinula edode, and Pleurotus ostreatus, the result suggested to differentiate Rhizoctonia solani AG1-IA in Taiwan or Japan and other AGs in similarity 68.8%; and further, isolates of Rhizoctonia solani AG1-IA from Taiwan could differentiate five groups in similarity 86%. Group north: isolates from Yilan, Hsinchu, Taoyua. Group midland: isolates from Taichung, hanghua, Nantou, Chiayi, Yunlin. Group east: isolates from Hualien. Group south: Pingtung, Tainan, Chiayi. Group complex: a large breadth group. The relationship between MCGs and dendrogram based on SSR-PCR analysis showed that diversity of isolates from Taiwan had high degree with geography. This had a great effect upon the diversity of isolates from Taiwan with geography morr than with rice variety (host). In the future, to collect isolates abidingly from Taiwan and to establish the database about rice sheath blight pathogen Rhizoctonia solani AG1-IA. And then, using the database to apply in rice control and breeding.
其他識別: U0005-2508200602174600
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