基于线粒体序列特征的茶树菇及杨柳田头菇分离物快速鉴定

The Fast Identification of the Isolation of Two Confusing Species A.aegerita and A.salicacola Based on the Mitochondrial Sequences

为探寻特异的DNA序列,快速鉴定茶树菇(Agrocybe aegerita)及其形态上易混淆种,给种质资源收集及利用提供有效工具。以云南采集到的18个菌株为材料,其中5株为A.aegerita,13个为A.salicacola,并对线粒体小亚基V9区进行序列分析。结果发现A.aegerita比A.salicacola多1个约50碱基的插入片段,而其他序列组成一致。根据此特点,在插入区筛选到1个特异DraⅠ酶切位点。通过对18个菌株V9区PCR产物进行酶切,发现A.aegeritaV9片段能完全被切开,而A.salicacolaV9片段则为完整。根据此酶切方法,可以高效鉴定A.aegerita及A.salicacola2个近似种,为食用菌遗传及育种研究提供了重要的基础。

The aim was to explore special DNA sequences,and identify the confusing species of A.aegerita in morphology for providing an efficient tool for the resource collection and utilization.In this study,18 strains including 5 A.aegerita and 13 A.salicacola collected in Yunnan Province were investigated on their small mitochondrial V9 sequences.The result showed the identical sequences were found in 2 species except one insertion segment in A.aegerita compared with A.salicacola.Based on the sequence characterization,one unique enzyme digestion site of DraⅠwas found in insertion segment,and the verification on the V9 PCR products of 18 strains confirmed that segments of A.aegerita were digested,and converse result occurred in that of A.salicacola.According to the method of enzyme digestion,2 species could be efficiently distinguished,and it provided an important basis for genetics and breeding of edible mushroom.