橡胶灵芝菌（Ganoderma pseudoferreum）DNA 条形码检测 筛选

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 trnL DNA 外，其他 8 对引物均扩增得到 PCR 产物，产物大小范围为 268~1 355bp，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。

Ganoderma pseudoferreum Identification Using ITS1/ITS4 DNA Barcode Sequences

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.