一种白肉灵芝的快速鉴定方法  

A Rapid Identification Method of Ganoderma leucocontextum

作　　者：黄龙花[1] 贺凤[1] 刘远超[1] 黄志勇 钟元茂 史钏 胡惠萍[1] 

机构地区：[1]广东省微生物研究所省部共建华南应用微生物国家重点实验室/广东省菌种保藏与应用重点实验室,广州510070 [2]广东粤微食用菌技术有限公司,广州510070

出　　处：《农业生物技术学报》2017年第1期159-164,共6页Journal of Agricultural Biotechnology

基　　金：广东省科技计划农业攻关项目(No.2012A020100010);广东省科技计划项目(No.2015A030302055)

摘　　要：白肉灵芝(Ganoderma leucocontextum)栽培中,菌种的真实性问题一直是制约其栽培成功率的主要因素之一,为了快速准确地鉴定白肉灵芝,本研究对白肉灵芝、灵芝(赤芝,G.lingzhi)、紫芝(G.sinense)、漆光灵芝(亮盖灵芝,G.lucidum)、四川灵芝(G.sichuanense)、日本灵芝(G.japonicum)、树舌(G.applanatum)、有柄树舌(G.gibbosum)、弯柄灵芝(G.flexipes)、无柄灵芝(G.resinaceum)和紫铜灵芝(G.cupreum)等共17个灵芝属样品进行了ITS序列测定分析比对,从而发现白肉灵芝ITS序列在120~140 bp以及450~470 bp处有多个特异碱基位点,基于这些特异碱基位点设计白肉灵芝的特异引物对BRLZ-F和BRLZ-R,结果表明,该特异引物对在58℃退火温度下特异性好,仅能扩增出白肉灵芝相应的长348 bp的特异片段。因此该特异引物对及方法可用于白肉灵芝的快速准确鉴定。Ganoderma leucocontextum is morphologically similar to certain species from its genus in nature.And the so-called distinguishing white flesh within its basidiocarp seemed non-unique according to Ganodermataceae sampling around Southern China. ITS region was reported a DNA barcode marker for macrofungi, a unique DNA marker is yet expected for species identification within genus or genus group. This article designed pair of specific PCR primers for G. leucocontextum identification. Mycelium samples of the 17 selected species phylogenetically closed to G. leucocontextum were harvested and their ITS were isolated and amplified, based on the universal primers. The alignment result showed multiple non-conservative sites located at 120~140 bp and 450~470 bp. Two oligo sequences, BRLZ-F and BRLZ-R, were designed to match the 2 differentiate fragments as the forward and reverse primers for G. leucocontextum PCR identification.BRLZ-F and BRLZ-R bind efficiently to the DNA templates at 58 ℃ in annealing and only a 348 bp fragment was capable for amplification. Confirmatory experiment illustrates BRLZ-F and BRLZ-R is effective for G.leucocontextum identification within the Ganoderma genus.

关 键 词：白肉灵芝 ITS序列 特异引物 快速鉴定 

分 类 号：S567.31[农业科学—中草药栽培]