农杆菌介导的刺芹侧耳遗传转化体系的建立  

Construction of the genetic transformation system of Pleurotus eryngii by Agrobacterium‐mediated method

作　　者：盛立柱 宋冰[2] 戴月婷[2] 李丹[2] 付永平[2] 李玉[2] 

机构地区：[1]吉林农业大学农学院,吉林长春130118 [2]吉林农业大学食药用菌教育部工程中心,吉林长春130118

出　　处：《菌物学报》2015年第4期653-661,共9页Mycosystema

基　　金：国家重点基础研究发展计划(2014CB138305);国家自然基金项目(31471926);省技厅项目(20140101149JC)

摘　　要：以刺芹侧耳菌丝球为受体,潮霉素(Hyg)为筛选标记,应用农杆菌介导法对刺芹侧耳菌丝进行了遗传转化研究。潮霉素敏感性测试结果表明,刺芹侧耳Hyg耐受浓度为50mg/L。农杆菌介导的刺芹侧耳菌丝最佳遗传转化体系为:菌液浓度OD600=0.6–0.7,侵染时间30–35min,共培养时间2d,侵染液和共培养培养基中乙酰丁香酮(AS)浓度为1mg/m L;经潮霉素抗性筛选、PCR鉴定和GUS活性的组织化学分析,表明外源基因GUS已转入到刺芹侧耳菌丝中,并获得表达。本实验成功地建立了稳定的农杆菌介导的刺芹侧耳遗传转化体系。The mycelial pellets of Pleurotus eryngii were used as receptor for transformation by Agrobacterium-mediated method, and the genetic transformation system of P eryngii was established. Sensitivity test showed that the tolerance concentration for P eryngii against hygromycin was 50mg/L. The optimum genetic transformation system for Agrobacterium-mediated P eryngii mycelia was OD6oo=0.6-0.7 of the bacterial concentration, 30-35min of infection duration, 2 days of the co-cultivation period and lmg/mL of acetosyringone （AS） concentration. The results of PCR identification and the histochemical analysis of 13-glucuronidase （GUS） activities indicated that the exogenous gene gus was transferred into the mycelia of P. eryng/i by hygromycin resistance selection. A stable Agrobacterium-mediated genetic transformation system of P eryngii was constructed successfully

关 键 词：刺芹侧耳 菌丝球 农杆菌 GUS染色 转化体系 

分 类 号：S646[农业科学—蔬菜学]