Structural and Functional Analysis of Linear Plasmid DNAs of Plant Pathogenic Fungus

植物病原真菌线性质粒 DNA 的结构和功能分析

基本信息

批准号：
63480039
负责人：
HAYASHIDA Teruyoshi
金额：
$ 4.42万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480039/
关键词：
Plasmid of plant pathogenic fungus Rhizoctonia solani pRS64 plasmid Hairpin loops structure Sequence homology among plDNAs Chromosomal DNA 病原糸状菌の線状プラスミド R__ー.<solani>___ー菌の転移因子染色体DNAとpRS64との相同性 Rhizoctonia Solani菌 PRS64プラスミド巨大プラスミド

项目摘要

1. Plasmid-like DNA (plDNA) was found in 48 out of 114 field isolates of Rhizoctonia solani. These 48 isolates were distributed as follows among the nine anastomosis groups (AG) and intraspecific groups (ISG) recognized among Japanese isolates of R. solani : 0 in AG-1 (sasakii type), 1 in AG-1 (web-blight type), 0 in AG-2-1, 11 in AG-2-2 (rush type), 10 in AG-2-2 (root rot type), 10 in AG-3, 8 in AG-4, 4 in AG-5 and 4 in AG-6. Each isolate carried one, two or three plDNAs identified by gel electrophoresis. Electron microscopic analysis revealed that all these plDNAs were linear molecules. The sequence homology among plDNAs found in representative isolates was examined by Southern blot analysis, using nick-translated plDANs as probes. Considerable sequence homology was observed among plDNAs obtained from isolates within the same AG and ISG. The plDNAs occurring in the isolates of AG-2-2 were classified into two groups on the basis of the sequence homology.2. Three linear DNA plasmids were found in isolate RI-64 of anastomosis group 4 (AG-4) of Rhizoctonia solani. These plasmids, designated pRS64-1, -2, and -3, possessed the same size of 2.7 kb. Restriction mapping and Southern hybridization anlysis of pRS64-1, -2, and -3 revealed the presence of homologous regions at both termini. The plasmid DNAs were resistant to both 3'-exonuclease and 5'-exonuclease even after treatment with protease K or alkali. The length of both terminal fragments that were generated by restriction endonuclease digestion was doubled under the denaturation condition, indicating that the linear plasmid DNAs have hairpin loops at both termini. Southern blotting analysis of total DNA showed the presence of two types of dimeric forms of pRS64 DNA. One is a head-to-head dimer and the other is a tail-to-tail dimer. The role of these unique DNA structures in replication of th

1。质粒样DNA（PLDNA）在114个田杆菌的田间分离株中有48个发现。这48个分离株分布在以下分布中，如下9个吻合组（Ag）和种内组（ISG）在Ag-1（Sasakii型）的日本分离株中识别的种群内（ISG）（ISG）（sasakii型），Ag-1中的1个（Web-Blight类型）），ag-2-1中的0，在AG-2-2中为11，Ag-2-2（刺激类型），AG-2-2中的10（root ROT类型），Ag-3中的10，Ag-4中的8，4，4，Ag中的4 AG-6中的-5和4。每个分离株携带一个，两个或三个PLDNA，通过凝胶电泳鉴定。电子显微镜分析表明，所有这些PLDNA都是线性分子。使用尼克翻译PLDAN作为探针，通过Southern印迹分析研究了在代表性分离株中发现的PLDNA之间的序列同源性。在同一Ag和ISG中的分离株中获得的PLDNA中观察到了相当大的序列同源性。根据序列同源性，将发生在AG-2-2分离株中的PLDNA分为两组2。在根瘤菌4（AG-4）的分离株RI-64中发现了三个线性DNA质粒。这些质粒称为PRS64-1，-2和-3，具有相同的2.7 kb的大小。 PRS64-1，-2和-3的限制映射和南部杂交驱虫揭示了两个末端的同源区域。即使在用蛋白酶K或碱处理后，质粒DNA也对3'-核酸酶和5'-核酸酶都具有抗性。在变性条件下，由限制性核酸内切酶消化产生的两个末端片段的长度均加倍，表明在两个末端，线性质粒DNA都有发夹环。总DNA的Southern印迹分析显示了两种类型的二聚体形式的PRS64 DNA。一个是头对头二聚体，另一个是尾巴二聚体。这些独特的DNA结构在复制中的作用