农杆菌介导的刺芹侧耳遗传转化体系的建立

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/mL;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。

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

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 OD₆₀₀=0.6-0.7 of the bacterial concentration, 30-35min of infection duration, 2 days of the co-cultivation period and 1mg/mL of acetosyringone (AS) concentration. The results of PCR identification and the histochemical analysis of β-glucuronidase (GUS) activities indicated that the exogenous gene gus was transferred into the mycelia of P. eryngii by hygromycin resistance selection. A stable Agrobacterium-mediated genetic transformation system of P. eryngii was constructed successfully.