Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/20665
標題: 立枯絲核菌第四融合群致病性與DNA變異之探討
Exploration of pathogenicity and DNA variation of Rhizoctonia solani AG-4
作者: 黃柏挺
Huang, Po-Ting
關鍵字: Rhizoctonia solani
立枯絲核菌
random amplified polymorphic DNA (RAPD)
rDNA
riboprinting
pathogenicity
致病性
逢機增幅多型性核酸
核糖指紋
出版社: 植物學系
摘要: 立枯絲核菌 (Rhizoctonia solani) 為一世界性分佈之土媒植物病原真菌 ,因各地環境不同,其生長條件、型態、寄主植物及致病性都有明顯的差 異。本論文為探討此菌之致病性與DNA 變異,選取15株台灣地區之立枯絲 核菌第四融合群 (R. solani AG-4) 菌株,及一HGI 標準菌株,共16株供 試菌株。測試供試菌株接種葉萵苣幼苗之致病能力,結果顯示上述供試菌 株有8株強致病性菌株、2株中致病性菌株、4株弱致病性菌株、2株無致病 性菌株。篩選出12條逢機增幅多型性核酸 (RAPD) 之引子,增幅之產物經 洋菜凝膠電泳分析,記錄成二元 (binary) 資料,共獲得467個數據列, 以NTSYS-PC軟體進行分析,顯示強致病性菌株具有高相似度。以真菌通用 (universal) 核糖核酸引子ITS1與ITS4 進行PCR,增幅之ITS1-5.8S rDNA-ITS2之片段大小約 650至750 bp,再分別以內切限制EcoR I、Hae III、Mbo I等切割,產生多種核糖指紋 (riboprinting) 圖譜。ITS1-5.8 S rDNA-ITS2片段大小與核糖指紋圖譜與供試菌株之致病性具有相關性。
Rhizoctonia solani K(hn is a world-wide soil-borne pathogenic fungus which show tremendous variation in characteristics such as geographic locations, morphology, host specificity and pathogenicity. The relation of pathogenicity and DNA variation of fifteen isolates of Rhizoctonia solani belonging to anastomosis group (AG) 4 from Taiwan and one isolate belonging to HGI were examined. The virulence of each strain was tested in loose leaf lettuce seedlings. The results indicated that eight strains were designated hypervirulent, two strains were middle virulent, four strains were hypovirulent and two strains were non-virulent of the sixteen tested strains. The genetic variation of these strains were evaluated by random amplified polymorphic DNA (RAPD) method with a set of twelve primers. The amplification results revealed scorable polymorphisms among these isolates, and a total of 467 band positions were scored with binary coding and assayed for Numerical Taxonomy and Multivariate Analysis System (NTSYS) software. The results of analysis indicated that the DNA banding patterns of the highly virulent isolates were highly similarity each other. Approximate 650 to 750 base pairs of ITS1-5.8S rDNA ITS2 fragments were amplified by universal primers ITS1 and ITS4 of fungal ribosomal DNA. The fragments were subjected to restriction digestion with recognition endonucleases EcoRI, HaeIII, MboI and different riboprinting patterns were displayed. The virulence of the sixteen isolates can be related with differences in the size of the fragments of ITS1-5.8S rDNA ITS2 and riboprinting patterns.
URI: http://hdl.handle.net/11455/20665
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