影响农杆菌介导草菇遗传转化的因素研究  

Assessment of Factors Affecting the Transformation Efficiency of Volvariella volvacea Mediated by Agrobacterium tumefaciens

作　　者：王杰[1] 林俊芳[1] 郭丽琼[1] 

机构地区：[1]华南农业大学食品学院,广东广州510642

出　　处：《华南农业大学学报》2010年第1期38-41,共4页Journal of South China Agricultural University

基　　金：广东省科技计划项目(2008A020100024;2009B020201012);"十一五"国家科技支撑计划重点项目(2008BADA1B02);国家自然科学基金(30871768);华南农业大学校长基金(5100-K08197)

摘　　要：以草菇Volvariella volvacea菌株V51为受体材料,用农杆菌Agrobacterium tumefaciens介导法转入抗冷冻蛋白基因(afp),探索不同的转化受体、农杆菌种类、乙酰丁香酮(AS)的浓度、共培养的温度、时间等因素对转化效率的影响,并采用PCR及Southern杂交检测afp基因是否整合到草菇的基因组中.结果表明:以D660 nm为0.15的EHA105为侵染的农杆菌菌株,加入200μmol/L AS进行预培养,侵染菌丝球后转入含有AS为200μmol/L的IMA培养基上,28℃,共培养60 h为最佳转化条件.PCR与Southern杂交结果证明afp基因已整合到草菇的基因组中.afp gene was transformed into VolvarieUa volvacea via Agrobacterium tumefaciens. We evaluated some critical factors influenced transfer efficiency, including explant, Agrobacterium strain, concentration of acetosyringone （AS）, temperature and time of co-culture. The integration of afp gene into V. volvacea genome was determined by PCR and Southern blot. The resuhs showed that effective transformation was acquired when mycelium pellets inoculated with A. tumefaciens strain EHA105 with a optical density at D660nm of 0. 15, pre-cuhure with 200 umol/L AS, co-cultivation for 60 h at 28 ℃on IMA medium containing 200 umol/L AS. afp gene had been introduced into V. volvacea with Agrobacterium-mediated transformation successfully.

关 键 词：草菇 农杆菌 遗传转化 转化效率 

分 类 号：S646.13[农业科学—蔬菜学]