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Linear plasmid DNAs of the plant pathogenic fungus Rhizoctonia solani with unique terminal structures
Authors:Shun-ichiro Miyashita  Hirohiko Hirochika  Joh-E Ikeda and Teruyoshi Hashiba
Institution:(1) Laboratory of Plant Pathology, Faculty of Agriculture, Tohoku University, 981 Sendai, Japan;(2) Laboratory of Molecular Genetics, National Institute of Agrobiological Resources, 305 Tsukuba, Japan
Abstract:Summary 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 analysis of pRS64-1, -2, and -3 revealed the presence of homologous regions at both termini. The plasmid DNAs were resistant to both 3prime-exonuclease and 5prime-exonuclease even after treatment with proteinase 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 the plasmids is discussed.
Keywords:Fungus  Rhizoctonia solani  Linear plasmid  Hairpin loop structure  Dimeric form
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