Inter- and intraspecific variation in the ITS region of rDNA of ectomycorrhizal fungi in Fennoscandia as detected by endonuclease analysis
Inter- and intraspecific variation in the ITS region of rDNA of ectomycorrhizal fungi in Fennoscandia as detected by endonuclease analysis
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DOI：
10.1046/j.1469-8137.1997.00742.x
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发表时间：
1997-06-01
期刊：
影响因子：
9.4
通讯作者：
Nylund, JE
中科院分区：
文献类型：
--
作者：
Karen, O;Hogberg, N;Nylund, JE
采用限制性片段长度多态性（RFLP）方法，研究了17属44种外生菌根真菌rDNA内转录间隔区（ITS）的种间和种内变异。对于每个物种，两到五个标本凭证（主要是担子果），收集整个Fennoscandia，进行了检查。利用聚合酶链反应（PCR）和通用引物ITS1和ITS4扩增ITS区，并用三种内切酶进行RFLP分析。在ITS区的种内多态性被发现在7个物种（在9的132个标本馆凭证）。多态性是由于长度突变，范围从5到15 bp的7个多态性物种和突变的核酸内切酶限制性位点在6个物种，主要影响只有一个核酸内切酶，但在两个物种两个核酸内切酶。使用一个单一的内切酶，一个独特的RFLP模式，可以获得超过一半的调查物种。通过结合不同的核酸内切酶，34（77%）的物种可以区分。其余的RFLP类型发生在一个属。低内，但高种间的ITS区域的变化的基础上，它的结论是，大多数外生菌根形成的44个调查的物种应承认通过与此数据集的比较，如果菌根是从一个网站位于芬诺斯堪的纳维亚采样。然而，在包含ITS区域种内变异程度较高的更大地理区域的数据集中，或者当来自彼此远离的几个地点的菌根进行比较时，可能需要包括当地参考种。
Interspecific and intraspecific Variation in the internal transcribed spacer (ITS) region of rDNA of ectomycorrhizal fungi of 44 species in 17 genera were examined using the restriction fragment length polymorphism (RFLP) method. For each species, two to five herbarium vouchers (mainly basidiocarps), collected throughout Fennoscandia, were examined. The ITS region was amplified using the polymerase chain reaction (PCR) and the universal primers ITS1 and ITS4, and subjected to RFLP analysis with three endonucleases. Intraspecific polymorphisms in the ITS region were found in seven species (in nine of the 132 herbarium vouchers). Polymorphisms were due to length mutations, ranging from 5 to 15 bp in four of the seven polymorphic species and mutations in endonuclease restriction sites in six species, mostly affecting only one endonuclease, but in two species two endonucleases. Using a single endonuclease, a unique RFLP pattern could be obtained for more than half the investigated species. By combining different endonucleases, 34 (77%) of the species could be distinguished from another. The remaining RFLP types occurred in one genus. On the basis of the low intra- but high interspecific variation in the ITS region, it is concluded that most ectomycorrhizas formed by the 44 investigated species should be recognized by comparison with this dataset, if the mycorrhizas are sampled from a site located in Fennoscandia. However, in datasets from even larger geographical areas encompassing a higher degree of intraspecific variation in the ITS region, or when mycorrhizas from several sites distant from each other are compared, it might be necessary to include local reference species.