Identification of the Dry Rot Fungus, Serpula lacrymans, and the Wild Merulius, S. himantioides, by Amplified Ribosomal DNA Restriction Analysis (ARDRA)
Identification of the Dry Rot Fungus, Serpula lacrymans, and the Wild Merulius, S. himantioides, by Amplified Ribosomal DNA Restriction Analysis (ARDRA)
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通过扩增核糖体 DNA 限制性分析 (ARDRA) 鉴定干腐真菌 Serpula lacrymans 和野生 Merulius S. Himantioides
DOI：
10.1515/hf.1999.020
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
U. Moreth
中科院分区：
文献类型：
--
作者：
O. Schmidt;U. Moreth
总结干腐真菌Serpula lacrymans和形态相似的野生merulius S. himantioides，通过扩增核糖体DNA限制性内切酶分析（ARP-RFLP），证明该方法可作为室内重要经济真菌的诊断工具。该技术使用聚合酶链反应（PCR）扩增ITS区的相对可变序列，该序列位于核核糖体DNA（rDNA）重复单元的18 S和28 S RNA基因的高度保守部分之间。随后用限制性内切酶消化扩增子可能会在种和亚种水平上表现出差异。用通用引物ITS 1/ITS 4组合对所有菌株的ITS区进行了扩增. lacrymans和S. himantioides扩增。琼脂糖凝胶电泳结果显示，两种细菌的扩增产物大小均在630 bp左右。分别用内切酶对HhaI/HhaI和AvaII/MboII消化扩增子，发现两种菌株的rDNA-ITS片段相同，表明它们的遗传关系。另一方面，分别用Bgl I/Hinf I和Hae III/TaqI消化，通过不同的片段模式分离真菌。因此，ARDRA-ITS被证明是适合于这两种真菌的鉴定。
Summary Isolates of the dry rot fungus, Serpula lacrymans, and of the morphologically similar wild merulius, S. himantioides, were investigated by amplified ribosomal DNA restriction analysis (ARDRA) of the internal transcribed spacer (ITS) region to prove this method as diagnosis tool for the economically important indoor rot fungi. The technique uses the polymerase chain reaction (PCR) to amplify the relatively variable sequences of the ITS region arranged between the highly conserved portions of the 18S and 28S RNA genes of the nuclear ribosomal DNA (rDNA) repeat unit. Subsequent digestion of the amplicon with restriction endonucleases may exhibit differences at species and subspecies level. Using the universal ITS 1/ITS 4 primer combination, the ITS region of all isolates of S. lacrymans and S. himantioides was amplified. The size of the amplified products was about 630bp in both species, as estimated from agarose gel electrophoresis. Digestion of the amplicon with the endonuclease pairs AluI/HhaI and AvaII/MboII, respectively, revealed identical rDNA-ITS fragments for the isolates of both species, indicating their genetic relationship. On the other hand, digestion with BglI/Hinf I and HaeIII/TaqI, respectively, separated the fungi by means of different fragment patterns. Thus, ARDRA-ITS proved to be suited for the identification of both fungi.