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A new multilocus approach for a reliable DNA-based identification of Armillaria species.

机译:一种新的多基因座方法,可对基于蜜环菌的物种进行可靠的基于DNA的鉴定。

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In this paper we highlight and critically discuss limitations to molecular methods for identification of fungi via the example of the basidiomycete genus Armillaria. We analyzed a total of 144 sequences of three DNA regions commonly used for identifying fungi (ribosomal IGS-1 and ITS regions, translation elongation factor-1 alpha gene) from 48 specimens of six Armillaria species occurring in Europe (A. cepistipes, A. ostoyae, A. gallica, A. borealis, A. mellea, A. tabescens). Species were identified by comparing newly obtained sequences with those from the NCBI database, phylogenetic analyses and PCR-RFLP analyses of the three regions considered. When analyzed separately, no single gene region could unambiguously identify all six Armillaria species because of low interspecific and high intrasequence variability. We therefore developed a multilocus approach, which involves the stepwise use of the three regions. Following this scheme, all six species could be clearly discriminated. Our study suggests that, to improve the reliability of DNA-based techniques for species identification, multiple genes or intergenic regions should be analyzed.Registry Number/Name of Substance 0 (DNA, Fungal).
机译:在本文中,我们将以担子菌属蜜环菌属为例,重点并批判性地讨论鉴定真菌的分子方法的局限性。我们分析了来自欧洲(A. cepistipes,A.)的6种蜜环菌物种的48个标本中通常用于鉴定真菌的3个DNA区(核糖体IGS-1和ITS区,翻译延伸因子-1α基因)的144个序列。 ostoyae,A。gallica,A.boralis,A。mellea,A。tabescens)。通过将新获得的序列与来自NCBI数据库的序列,系统发育分析和所考虑的三个区域的PCR-RFLP分析进行比较,从而确定了物种。单独进行分析时,由于低的种间差异和高的序列内变异性,没有一个单一的基因区域可以明确识别所有六个蜜环菌属。因此,我们开发了一种多位置方法,其中涉及三个区域的逐步使用。按照这一计划,可以对所有六个物种进行明确区分。我们的研究表明,为了提高基于DNA的物种识别技术的可靠性,应分析多个基因或基因间区域。物质0的注册号/名称(DNA,真菌)。

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