7种镰刀菌引起头疫病的线粒体dna限制性内切片段长度多态性。

N. Kodo, C. Nakamura, H. Kato, Takumi Yoshiawa, N. Mori, C. Kaneda
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引用次数: 15

摘要

采用CsCl/双苯并亚胺密度梯度超离心技术,从小麦和其他谷物中引起枯萎病和霉菌毒素污染的7种镰刀菌21株中纯化了线粒体dna (mtdna)。利用F. graminearum菌株KU-1615的mtDNA构建了12个PstI片段中缺失1个(14.7 kb)的部分PstI克隆库。7种代表性菌株经4或5种限制性内切酶(BamHI、MluI、PstI、pvii和XhoI)单酶切、双酶切和三酶切测定mtdna的分子量,其余菌株经BamHI和/或PstI单酶切测定mtdna的分子量。不同菌株的MtDNA大小不同,最小的为一株avenaceum的49 kb,最大的为一株culmorum的116 kb。限制性内切片段长度多态性(RFLP)分析显示,种间差异较大,所有种均根据其限制性内切片段模式进行鉴定,并将其划分为单独的聚类,但tricinctum菌株的限制性内切片段模式与孢子毛线虫的两种菌株中的一种相同。还发现了包括大小变异在内的相当大的种内变异。这些结果表明,种间和种内的插入/缺失发生率很高。在聚类分析的基础上,讨论了镰刀菌属植物的分类问题。
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Restriction fragment length polymorphisms of mitochondrial DNAs from seven Fusarium species causing Fusarium head blight.
Mitochondrial DNAs (mtDNAs) were purified by CsCl/bisbenzimide density-gradient ultracentrifugation from 21 strains of seven Fusarium species that cause fusarium head blight and mycotoxin contamination in wheat and other cereals. A partial PstI clone bank, from which one of twelve PstI fragments (14.7 kb) is missing, was constructed using mtDNA from strain KU-1615 of F. graminearum. Molecular sizes of mtDNAs of single representative strains from the seven species were determined after single-, double- and triple-digestion by four or five restriction enzymes (BamHI, MluI, PstI, PvuII and XhoI), while those of others were after single-digestion by BamHI and/or PstI. MtDNA size varied from the smallest 49 kb in one strain of F. avenaceum to the largest 116 kb in one strain of F. culmorum. Restriction fragment length polymorphism (RFLP) analysis revealed a large interspecific variation, thus all the species were identified by their restriction fragment patterns and assigned to individual clusters except for F. tricinctum in that a strain studied showed identical patterns to one of two strains of F. sporotrichioides. Considerable intraspecific variation including size variation was also detected. These results indicated a high incidence of insertions/deletions both between and within species. On the basis of results obtained by the cluster analysis, some aspects of taxonomy in these Fusarium species were discussed.
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