Clapp J P, Rodriguez A, Dodd J C
Research School of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, UK.
International Institute of Biotechnology/Research School of Biosciences (UKC), 1/13 Innovation Buildings, Sittingbourne Research Centre, Sittingbourne, Kent ME9 8HL, UK.
New Phytol. 2001 Mar;149(3):539-554. doi: 10.1046/j.1469-8137.2001.00060.x.
• High levels of variation are reported in the large subunit (LSU) rRNA gene, D2 region of Glomus coronatum, a well characterized species of arbuscular mycorrhizal fungus (AMF). • Clones (435) containing the D2 regions from 7 isolates of G. coronatum were investigated for intra- and inter-isolate sequence variation using PCR-single-strand conformational polymorphism (PCR-SSCP) as a prescreen before sequencing. Isolates of G. mosseae, G. constrictum and G. geosporum, three species of AMF with similar spore ontogeny and morphology, were also analysed. • Analysis of 138 representative sequences indicated that most were unique; this variation could not be attributed to DNA polymerase or cloning artefacts. Only 13 sequences were found in more than one isolate. Neighbour-joining analysis showed that most sequences from G. coronatum formed a main group although several sequences from G. mosseae and G. constrictum clustered with G. coronatum. • There was greater than expected variation in the LSU D2 region sequences from G. coronatum. The four Glomus species, closely related by spore morphology, might represent part of a genetic continuum. Implications for the concept of species in AMF, the use of rRNA sequences to estimate biodiversity and in situ detection in field ecology are discussed.
• 据报道,丛枝菌根真菌(AMF)中一种特征明确的物种——冠球囊霉(Glomus coronatum)的大亚基(LSU)rRNA基因的D2区域存在高度变异。
• 对来自7个冠球囊霉分离株的含有D2区域的435个克隆进行了研究,在测序之前,使用聚合酶链反应-单链构象多态性(PCR-SSCP)作为预筛选方法来检测分离株内和分离株间的序列变异。还分析了摩西球囊霉(G. mosseae)、缩球囊霉(G. constrictum)和地生球囊霉(G. geosporum)这三种具有相似孢子发育和形态的AMF物种。
• 对138个代表性序列的分析表明,大多数序列是独特的;这种变异不能归因于DNA聚合酶或克隆假象。只有13个序列在不止一个分离株中被发现。邻接法分析表明,尽管来自摩西球囊霉和缩球囊霉的几个序列与冠球囊霉聚类在一起,但冠球囊霉的大多数序列形成了一个主要类群。
• 冠球囊霉的LSU D2区域序列存在大于预期的变异。这四个通过孢子形态密切相关的球囊霉物种可能代表了遗传连续体的一部分。文中讨论了其对AMF物种概念、利用rRNA序列估计生物多样性以及野外生态学原位检测的影响。