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通过两种基于PCR的分型方法评估木耳菌株的遗传多样性。

Assessment the genetic diversity of Auricularia strains by two PCR-based typing methods.

作者信息

Wen Ya-Li, Cao Hui, Pan Ying-Jie

机构信息

Department of Microbiology, Nanjing Agricultural University, Nanjing 210095, China.

出版信息

Wei Sheng Wu Xue Bao. 2004 Dec;44(6):805-10.

Abstract

Two PCR-based methods, enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) and randomly amplified polymorphic DNA (RAPD), were adopted for differentiating Auricularia strains. Taken the similarity coefficient as 75%, 29 strains of three Auricularia species were grouped into 6 and 9 clusters by RAPD and ERIC, respectively. The dendrogram from ERIC exhibited two distinct parts, one representing A. auricula and the other A. polytricha, but the dendrogram from RAPD failed to clearly distinguish between these two species. However, both methods similarly revealed high homology between A. fuscosuccinea and A. auricula. The homology relationships among the three species obtained from ERIC were validated by Southern hybridization. The analyses showed that RAPD is able to differentiate mainly at the species level, while ERIC is effective at the strain level and therefore more consistent with cultivation characteristics. The results indicate that the method of ERIC-PCR is more rapid and reliable than RAPD, and may substitute for RAPD in research related to the genetic identification and genetic diversity in Auricularia.

摘要

采用两种基于聚合酶链式反应(PCR)的方法,即肠杆菌重复基因间共有序列聚合酶链式反应(ERIC-PCR)和随机扩增多态性DNA(RAPD),对木耳菌株进行鉴别。以相似系数75%为标准,三种木耳属菌株的29个菌株分别通过RAPD和ERIC-PCR被聚为6个和9个聚类群。ERIC-PCR的聚类图呈现出两个明显的部分,一部分代表黑木耳,另一部分代表毛木耳,而RAPD的聚类图未能清晰区分这两个物种。然而,两种方法均同样显示出皱木耳与黑木耳之间具有高度同源性。通过Southern杂交验证了从ERIC-PCR获得的这三个物种之间的同源关系。分析表明,RAPD主要能够在物种水平上进行鉴别,而ERIC-PCR在菌株水平上有效,因此更符合栽培特性。结果表明,ERIC-PCR方法比RAPD更快速可靠,在木耳的遗传鉴定和遗传多样性相关研究中可能会取代RAPD。

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