小菇一种Mycena sp.的DNA提取及测序

DNA Extraction and Sequencing of One Species of Mycena sp.

[目的]为新野生真菌小菇一种Mycenasp.的分类提供依据。[方法]采集到一种新的野生真菌,经形态学观察初步确定为小菇一种Mycenasp.。对该真菌进行了DNA提取、18SrDNA的PCR扩增与产物回收、琼脂糖凝胶电泳检测和测序等一系列分子生物学研究,对其基因序列进行了Blast比对和同源性数据检索。[结果]利用通用引物进行18SrDNA的PCR扩增,可获得约1.8 kb的扩增片段。通过序列分析,对回收后的产物进行鉴定,确定其为18SrDNA基因序列。通过对小菇一种Mycenasp.的18SrDNA基因序列进行Blast比对和同源性数据检索,发现其与白蘑属、毛伞属和脐伞属的亲缘关系最近。[结论]可推测该野生真菌为担子菌亚门、层菌纲、伞菌目、白蘑科一种,并结合其形态特征,暂时确定为小菇一种Mycenasp.。

[Objective]The aim of the research was to provide basis for the classification of a new kind of wild fungi-one species of Mycena sp. [Method] A new kind of wild fungiwas collected and it was preliminarily confirmed as one species of Mycena sp. though motphological observation. A series of molecular biology researches such as DNA extraction, PCR amplification of 18S rDNA and products reclamation, agarose gel electrophoresis detection and sequencing were conducted on this epiphyte. And Blast analysis and homology data retrieval was made on the sequence of this gene. [Result] PER amplification of 18S rDNA was conducted by using universal primers and about 1,8 Ida amplification fragment was obtained. Through identifying the reclaimed products by sequence analysis, it was confirmed as 18S rDNA gene sequence. According to Blast analysis and homology data retrieval of 18 rDNA gene sequence of one species of Mycena sp., it was found it had the closest genetic relatioaship with Tricholoma mongolicum, Crinipellis sp. and Omphalia. [Conclusion] It can be predicted that this wild fungi was Basidiomycotina, Hymenomycetes, Agaricales, one species of T. mongolicum family. Combining with its motphological characteristics, it was confinmed as one species of Mycena sp. for the moment.