臺灣博碩士論文加值系統

水稻是許多國家的主要糧食，許多植物病原菌皆會危害水稻並降低其產量，其中以水稻稻熱病、水稻紋枯病為主要病害。水稻紋枯病是由立枯絲核菌第一融合群亞群(Rhizoctonia solani AG1-IA)所引起，紋枯病菌主要發生在高溫高濕氣候的地區，由於台灣依年四季多為高溫多濕氣候所以病害發生快速而嚴重。因此藉由了解有關水稻紋枯病其遺傳差異性以及菌絲型態、生長型態以及菌核分佈等差異性並輔以病原菌毒力強弱的比較、即經由溫室水到接種後所產生之病害指數來比較菌株來源及其供試菌株採集地在地理區域上的差異性等比較並综合其差異以及相似因子進行分析以獲得菌株間的特質為一研究水稻紋枯病菌在台灣之多樣性模式。而經由與參考菌株進行菌絲融合以及使用Rho1以及Rho2所增幅的特殊片段能專一性的針對Rhizoctonia solani AG1-IA對於其他亞群作出區別的結果進行比較後發現，使用專性引子作為鑑定方式所得到的結果可以支持利用菌絲融合進行鑑定而鑑別出屬於Rhizoctonia solani AG1-IA的菌株。而利用隨機合成引子針對各地蒐集到之供試菌株以及參考菌株進行增幅的結果進行分析發現各地供試菌株於演化樹上之相似度大於48%時的叢集符合與之地緣相關的河川流域，而以dsRNA條帶上的特徵作為親緣性分析則是與地緣性並無相關並於相似度大於72%時大致分為八個叢集。另一方面、經由接種於三種水稻所統計的病害指數則發現整體病害嚴重的程度而言、台梗11號大於台梗9號大於台農71號。另一方面、對於供試菌株中dsRNA出現的種類則與菌株的生理生化特性以及供試菌株接種三種水稻所獲得之病害指數而言則也是無相關性。在本研究三項因子之相關性只有病害接種部分結果與供試菌株生理生化特性有相關性以及RAPD對於供試菌株分群的結果與灌溉水源流域有相關性之外其餘則較沒有相關性。對於水稻紋枯病菌在台灣之多樣性而言水稻紋枯病菌唯一具有區域相關性以及其生理生化特徵與病害發生強弱有相關之水稻病原菌，因此對於台灣水稻紋枯病菌之多樣性而言有長期探討之必要性。

Rice is used to be the dominated food in many countries, with the suffering of plant pathogen like rice blast and rice sheath blight, the production is reduced. Rice sheath blight caused by the pathogen of Rhizoctonia solani AG1-IA and the pathogen dominantly happens in the area with high temperature and moist weather. Besides, the weather above each season with humidity and high temperature facilitate the serious disease. Therefore, with the understanding of the genetic difference, characteristic, and the difference of disease severity of different test isolates, the process to discriminate the similarity and the resource of each test isolates collected from, to have the idea of condition of rice sheath blight in Taiwan could became possible. Besides, after the recognition of anastomosis test and the specific primer Rho1 and Rho2 designed to detect the specific sequence of test isolates, the result of molecule tool totally similar with the anastomosis test. That means the molecule tool can separate the difference between different groups of Rhizoctonia solani. Besides, with the composeion of the information with RAPD of 13 primers, the cluster of discrimination with phylogenetic tree of each test isolates and reference isolates are recognized in to 8 clusters at the simileruty of 48%. The clusters with phylogenetic tree of dsRNA are divided into also 8 clusters at similarity of 72%. On the otherhand, with the calculation of diseasivity after the inoculation with rice culture in greenhouse, the disease severity is subsequently worse form Tai-nung No. 71 to Tai-keng No.9 and to Tai-keng No. 11. Besides, there is no relativity of the elements of the quantity, hypha growth rate and the radius of the distribution of sclerotium of test isolates pretend to be positive related. And the existing of dsRNA seems to have nothing positively related with the disease severity of inoculation of test isolates and the phenomenon with the culture of test isolates. The conclusion is that the genetic characteristic of test isolates are related with the geographical resource of water irrigation is the valuable result of this research.

目錄
壹、 中文摘要 4
貳、 英文摘要 6
参、 前言 8
肆、 材料與方法 18
一、 供試水稻紋枯病菌來源 18
1 供試水稻紋枯病菌的收集、分離、培養 18
2 供試水稻紋枯病菌鑑定 18
二、 供試水稻紋枯病菌生長勢與培養型態測試 19
1 供試水稻紋枯病菌菌落型態 19
2 供試水稻紋枯病菌菌核產生數量 19
3 供試水稻紋枯病菌菌絲生長速率 19
4 供試水稻紋枯病菌菌核分佈型態 19
三、 總量去氧核醣核酸(Total DNA)的抽取及其濃度測定 20
1 供試水稻紋枯病菌菌絲培養 20
2 總量DNA的萃取以及濃度測試 20
3 水稻紋枯病菌專一性引子鑑定 21
四、 雙股核醣核酸(dsRNA)的抽取 22
1 水稻紋枯病菌培養 22
2 雙股核醣核酸(dsRNA)的抽取 22
五、 供試水稻於溫室中之接種 23
1 台梗11號、台梗9號以及台農71號水稻之溫室栽種 23
2 供試水稻紋枯病菌接種 24
3 供試水稻紋枯病菌接種後之病害指數測定 24
4 供試水稻紋枯病菌接種後之病徵高度 24
六、 隨機引子增幅核酸多型性(RAPD)各產物之表現 24
1 RAPD增幅反應 25
2 RAPD增幅產物親緣性之分析 25
伍、 結果 27
陸、 討論 32
柒、 表 37
捌、 圖 48
玖、 文獻 71

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