用于检测两种古老的内生菌根真菌——隐匿球囊霉和巴西球囊霉的分类群特异性寡核苷酸引物。

Taxon-specific oligonucleotide primers for detection of two ancient endomycorrhizal fungi, Glomus occultum and Glomus brasilianum.

作者信息

Millner P D, Mulbry W W, Reynolds S L

机构信息

US Department of Agriculture, Agricultural Research Service, Beltsville Agricultural Research Center, Sustainable Agricultural Systems Laboratory, 10300 Baltimore Blvd, Bldg. 001, Rm140, Beltsville, MD 20705-2350, USA.

出版信息

FEMS Microbiol Lett. 2001 Mar 15;196(2):165-70. doi: 10.1111/j.1574-6968.2001.tb10559.x.

DOI:10.1111/j.1574-6968.2001.tb10559.x

PMID:11267774

Abstract

A unique oligonucleotide pair, GOCC56:GOCC427, was designed that correctly primed specific amplification of a approximately 370-bp sequence spanning the ITS and 5.8S rDNA regions of Glomus occultum and Glomus brasilianum. In addition, this primer pair successfully detected G. occultum and G. brasilianum DNA in nested PCR using a primary PCR product amplified from highly diluted extracts of colonized corn (Zea mays) roots using modified ITS1:ITS4 primers. A second primer pair, GBRAS86:GBRAS388, primed specific amplification of a approximately 200-bp sequence spanning the ITS and 5.8S rDNA regions present only in G. brasilianum and Glomus strain GR582. Combined use of both primer pairs provides the means to detect and differentiate two ancient endomycorrhizal species, G. occultum and G. brasilianum, undetectable by standard root staining procedures. Sequence analysis showed that the purported G. occultum strain GR582 is likely a strain of G. brasilianum.

摘要

设计了一对独特的寡核苷酸引物，即GOCC56:GOCC427，它能正确引发对球囊霉属隐匿种（Glomus occultum）和巴西球囊霉（Glomus brasilianum）的内转录间隔区（ITS）和5.8S核糖体DNA（rDNA）区域约370 bp序列的特异性扩增。此外，在巢式聚合酶链反应（PCR）中，使用从经改良的ITS1:ITS4引物扩增的定殖玉米（Zea mays）根的高稀释提取物中获得的初级PCR产物，该引物对成功检测到了隐匿球囊霉和巴西球囊霉的DNA。第二对引物GBRAS86:GBRAS388引发了仅存在于巴西球囊霉和球囊霉属菌株GR582中的ITS和5.8S rDNA区域约200 bp序列的特异性扩增。同时使用这两对引物提供了检测和区分两种古老的内生菌根真菌——隐匿球囊霉和巴西球囊霉的方法，而这两种真菌通过标准的根染色程序无法检测到。序列分析表明，所谓的隐匿球囊霉菌株GR582可能是巴西球囊霉的一个菌株。