丹参、黄精内生放线菌的分离及遗传多样性分析

Isolation and Genetic Diversity of the Endophytic Actinomycetes from Salvia miltiorrhiza Bge.and Polygonatum sibiricum Red.

从四川遂宁地区采集了丹参、黄精2种道地中药材,样品通过0.87%次氯酸钠不同时间梯度消毒,用组织法和匀浆法对植株进行处理,并在HV、G2、S等培养基中加入不同浓度的重铬酸钾和萘啶酮酸以抑制非放线菌的生长,确定了分离中药材内生放线菌比较适宜的方法。经分离、纯化得到52株菌落大小、形态、颜色各异的内生放线菌。选取其中12株代表菌株进行16SrRNAPCR-RFLP分析,在88%的相似水平上,被分为5个遗传类型,表明了药用植物内生放线菌的遗传多样性。

Two species of Chinese medicinal plants,Salvia miltiorrhiza Bge.and Polygonatum sibiricum Red.,were collected from Suining region of Sichuan province.The plants were disinfected with 0.87% hypoclorite under different time and treated with the methods of organic law and homogenate law,respectively.Then endophytes were isolated in the media HV,G2,S amended with which use different concentrations of potassium dichromate plus malidixic acid as inhibitors.The method of isolating endophytic actinomycetes from the medicinal plants was optimized.Fifty-two endophytic actinomycetes with different colony size,shape and color were isolated and purified.Twelve representative strains were chosen for genetic diversity study by 16S rRNA PCR-PFLP.Five clusters were abtained based the 16S rRNA PCR-RFLP pattens which indicated the high genetic diversity of the endophytic actinomycetes in medicinal plants.