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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 objective of this study was to investigate the genetic transformation of Pleurotus ostreatus with Agrobacterium tumefaciens mediated by hygromycin. Hygromycin sensitivity test results show that the celery side of the ear Hyg tolerance concentration of 50mg / L. Agrobacterium tumefaciens mediated the best genetic transformation system Pleurotus ostreatus for the bacteria concentration OD600 = 0.6-0.7, infection time 30-35min, co-culture time 2d, the infection solution and co-culture medium acetosyringone (AS) concentration of 1mg / m L; hygromycin resistant screening, PCR identification and histochemical analysis of GUS activity, indicating that the foreign gene GUS has been transferred to Pleurotus eryngii to obtain and express. The experiment successfully established a stable Agrobacterium-mediated E. sinensis genetic transformation system.