Establishment of the basis on molecular taxonomy of phytopathogenic fungi

植物病原真菌分子分类学基础的建立

• 批准号: 07660061
• 负责人: FURUYA Naruto
• 金额: $ 1.47万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660061/
• 关键词: Phizoctonia; Fatty acid analysis; 28SrDNA; PCR; RFLP; Rhizoctonia solani; リボソーム遺伝子; 菌体脂肪酸分析; 菌糸融合群; 植物病原糸状菌

Biochemical methods consisting of fatty acid analysis and restriction fragment length polymorphism (RFLP) analysis were performed for the objective indicator for identification and characterization of Rhizoctonia spp.and R.solani AGs.Percentage composition of whole-cellular fatty acids of 5 Rhizoctonia spp. (R.solani, R.oryzae, R.oryzae-sativae, R.fumigata and R.candida) and 7 AGs including 13 intraspecific groups (ISGs) of R.solani were characterized by gas-liquid chromatography. The major fatty acids identified were palmitic, palmitoleic, oleic and linoleic acids. Principal componet analysis based on the percentage composition of whole-cellular fatty acids revealed consistently low variability among isolates of a single species and significant differences among 5 Rhizoctonia spp.Similar results by principal component analysis were presented low variability among isolates of an identical AG and differences among seven AGs of R.solani. Average linkage cluster analysis also showed that … More the isolates in the same species of Rhizoctonia spp.and an identical AG of R.solani formed individual clusters on the dendrogram, and each species of Rhizoctonia and different AGs R.solani couls be distinguished based on the dendrogram. However, all of the ISGs belonging to the AG-1, AG-2 and Ag-4 isolates were differentiated by the average linkage cluster analysis. Based on percentage composition of fatty acids, 3ISGs in AG-1,5 ISGs in AG-2, and 2 ISGs in AG-4, were presented respectively. Isolates of R.solani AG-1 IA showed specific and significant composition of whole-cellular fatty acids from other AGs and ISGs of R.solani.Isolates of 5 Rhizoctonia spp.and 7 AGs representing 13 ISGs of R.solani were characterized by the RFLPs analysis of a 28S rDNA gene amplified by the polymerase chain reaction (PCR). Amplification products by PCR reaction were 1.4-and 1.8-kirobase pair fragments. RFLP profiles obtained after digestion of 28S rDNA with the restriction enzymes were different depending on the 4enzymes, MspI,HhaI,Sau3AI and HaeIII,used. The RFLP profiles by the digestion with MspI and Hhal showed specific restriction fragments polymorphism among 5 Rhizoctonia spp. Variations in the PCR-amplified rDNA products and the polymorphisms on digestion with 4 restriction enzymes were also observed among 7 AGs of R.solani. These differences were maily conseved among ISGs of AG-1 and AG-2. A dendrogram derived from the RFLP profiles showed that ISGs from AG-1 and AG-2 can each be subdivided into distinct groups, that are distantly related to the majority isolates of the other AGs. Isolates of R.solani AG-1 IA showed specific RFLP profiles and phylogenetic differences from the other ISGs and AGs of R.solani.These two nethods, fatty acid analusis and RFLP analysis, clearly concluded the significant difference among isolates of Rhizoctonia spp. Less

进行了由脂肪酸分析和限制片段长度多态性（RFLP）分析组成的生化方法，以用于鉴定和表征Rhizoctonia spp的客观指标。和r.solani ags。5根contonia spp的全细胞脂肪酸的百分比组成。 （R.Solani，R.Oryzae，R.OryZae-Sativae，R.Fumigata和R.Candida）和7 AGS和7 AGS，包括13个r.solani的13个种内组（ISG），以气体色谱法进行了表征。确定的主要脂肪酸是棕榈酸，棕榈油酸，油酸和亚油酸。基于整个细胞脂肪酸的百分比组成的百分比组成的主要成分分析表明，单个物种的分离株之间的变异性始终较低，并且在5个根瘤菌属的5个根源分析之间存在显着差异。主成分分析的类似结果显示了相同AG的分离株之间相同AG和R.Solani的七个Ags ags ags ag和差异的差异。平均连接群集分析还表明，……更多的分离株在同一种类的根瘤菌中，以及在树状图上形成的单个簇的相同Ag，并且根瘤菌和不同的Ags r.solani物种可以根据树状图来区分。但是，通过平均链接群集分析，属于AG-1，AG-2和AG-4分离株的所有ISG均分化。基于脂肪酸的百分比，AG-2中的Ag-1,5 ISG中的3ISG和AG-4中的2个ISGS分别提出。 Isolates of R.solani AG-1 IA showed specific and significant composition of whole-cellular fatty acids from other AGs and ISGs of R.solani.Isolates of 5 Rhizoctonia spp.and 7 AGs representing 13 ISGs of R.solani were characterized by the RFLPs analysis of a 28S rDNA gene amplified by the polymerase chain reaction (PCR).通过PCR反应扩增产物为1.4和1.8千里酶对片段。用限制酶消化28S rDNA后获得的RFLP曲线不同，具体取决于使用的4酶，MSPI，HHAI，SAU3AI和HAEIII。用MSPI和HHAL消化的RFLP谱图显示了5个Rhizoctonia spp之间的特定限制片段多态性。在R.Solani的7 AGS中，还观察到了4种限制酶消化的PCR放大rDNA产物和多态性的变化。这些差异是在AG-1和AG-2的ISG之间汇总的。从AG-1和AG-2的ISG中得出的源自RFLP轮廓的树状图可以分为不同的组，这些组与其他AGS的多数分离株遥远相关。 R.Solani Ag-1 Ia的分离株显示出与其他ISGS和AGS的特定RFLP谱和系统发育差异。这两种Nethods，脂肪酸止痛和RFLP分析清楚地结论了根瘤菌分离株之间的显着差异。较少的

项目成果

Masaru Matsumoto: "A study on fatty acid analysis as a new taxonomic tool for differentiating Rhizoctonia spp." J.Fac.Agr.,Kyushu Univ.140(3・4). 279-286 (1996)

Masaru Matsumoto：“脂肪酸分析作为区分丝核菌的新分类工具的研究。”J.Fac.Agr.，九州大学140（3・4）（1996）。

Masaru Matsumoto: "Characterization of Rhizoctonia spp.,causal agants of sheath diseases of Rice plant by whole cellular fatty acides analysis." Ann.Phytopath.Soc.Japan. 63・3(印刷中). (1997)

Masaru Matsumoto：“通过全细胞脂肪酸分析来表征丝核菌，水稻鞘病的致病菌。”Ann.Phytopath.Soc.Japan 63·3（出版中）。

Matsumoto, M., Furuya, N.and Matsuyama, N.: "PCR-RFLP analysis of amplified 28S ribosomal DNA for identification of Rhizoctonia spp., the causal agents of sheath diseases of rice plants" J.Fac.Agr., Kyushyu Univ.41(1・2). 39-44 (1996)

Matsumoto, M.、Furuya, N. 和 Matsuyama, N.：“扩增 28S 核糖体 DNA 的 PCR-RFLP 分析用于鉴定丝核菌，水稻鞘病的致病因子”J.Fac.Agr.，Kyushyu大学41（1·2）39-44（1996）

Matsumoto, M., Furuya, N., Takanami, Y.and Matsuyama, N.: "RFLP analysis of the PCR-amplified 28SrDNA in Rhizoctonia solani" Mycoscience. 37. 351-356 (1996)

Matsumoto, M.、Furuya, N.、Takanami, Y. 和 Matsuyama, N.：“立枯丝核菌中 PCR 扩增的 28SrDNA 的 RFLP 分析”Mycoscience。

Masaru Matsumoto: "Characterization of Rhizoctonia spp.,causal agents of sheath diseases of rice plant,by total cellular fatty acids analysis" Ann.Phytopathol.Soc.Jpn.63・3. 149-154 (1997)

Masaru Matsumoto：“通过总细胞脂肪酸分析来表征水稻纹枯病的致病菌”Ann.Phytopathol.Soc.Jpn.63・3（1997）。