摘要
选用10对DNA条形码标准序列ITS、COI、nrDNA-LSU、nrDNA-SSU和trnL DNA对海南和云南红根病区分离的21个橡胶灵芝菌进行检测。除COI和trnLDNA外,其他8对引物均扩增得到PCR产物,产物大小范围为268～1355bp,种内相似度为98.05%～99.54%。分别将扩增得到的序列与GenBank公布上的Ganoderma属和常见真菌属序列,共77个ITS、74个nrDNA-LSU和74个nrDNA-SSU序列进行系统进化树构建与分析。结果表明:nrDNA-LSU序列和nrDNA-SSU序列种间差异较小,不能准确区分Ganoderma属及其他属;5对ITS序列均能区分Ganoderma属,但仅ITS1/ITS4序列能将Ganoderma属下16个种归入不同分支。ITS1/ITS4序列表现出适宜的种内与种间差异,适宜作为G. pseudoferreum的DNA条形码。
This study was aimed to identify Ganoderma pseudoferreum by using ten pairs of DNA barcode sequences including COI, ITS, nrDNA-LSU, nrDNA-SSU and trnL. Twenty-one strains of G. pseudoferreum were obtained from fourteen rubber planting farms in Hainan and Yunnan provinces from May to July in 2016. DNA barcode standard sequences were tested by PCR and sequencing. Eight pairs of primers were amplified to obtain PCR products except COI and trnL DNA. The PCR product sizes ranged from 268 bp to 1 355 bp, and intra-specific similarity were 98.05%– 99.54%. Sequences of 77 ITS, 74 nrDNA-LSU and 74 nrDNA-SSU downloaded from GeneBank of Ganoderma and other common fungi were constructed and analyzed. The results indicated that the inter-specific differences between the nrDNA-LSU and nrDNA-SSU sequences were too small to distinguish Ganoderma and other genera accurately. All the five ITS sequences could distinguish Ganoderma, but only ITS1/ITS4 sequence could classify the 16 species of Ganoderma into different branches. ITS1/ITS4 sequences showed suitable intra and inter-specific differences and could be used as a suitable DNA barcode for fast identification of G. pseudoferreum.
作者
蔡海滨
涂敏
孙爱花
程汉
安泽伟
胡彦师
李维国
CAI Haibin;TU Min;SUN Aihua;CHENG Han;AN Zewei;HU Yanshi;LI Weiguo(Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Biology and Genetic Resources of Rubber Tree,Ministry of Agriculture / State Center for Rubber Tree Breeding,Danzhou,Hainan 571737,China)
出处
《热带作物学报》
CSCD
北大核心
2018年第11期2225-2234,共10页
Chinese Journal of Tropical Crops
基金
中国热带农业科学院橡胶研究所基本业务费项目(No.1630022015002)
中国热带农业科学院基本科研业务费项目(No.1630022017010)
农业农村部物种品种资源保护项目(No.2130135)
