鸭梨多酚氧化酶基因反义表达载体的构建及农杆菌介导的遗传转化  

Construction and Transformation for Antisense Expression Vector of Polyphenol Oxidase Gene in Yali Pear(Pyrus bretschneideri Rehd.)

作　　者：李桂琴[1,2] 齐靖[1] 高志华[2] 许冬倩[2] 李会宣[2] 张玉星[1] 

机构地区：[1]河北农业大学园艺学院,保定071001 [2]河北经贸大学生物科学与工程学院,石家庄050061

出　　处：《植物遗传资源学报》2010年第5期635-639,共5页Journal of Plant Genetic Resources

基　　金：河北省科技攻关计划项目(042401116D-2)

摘　　要：根据鸭梨多酚氧化酶基因序列设计引物,PCR扩增该基因3′端450bp的片段,并将该片段反向插入真核表达载体pB I121的CaMV 35S启动子和NOS终止子之间,首次构建了鸭梨PPO基因的反义表达载体;其后,在农杆菌EHA105的介导下,成功实现了PPO反义基因对鸭梨组培苗的遗传转化。经Northern杂交和酶活检测证实,转基因鸭梨植株体内的多酚氧化酶基因转录和翻译水平均得到明显抑制,从而为耐褐化梨新品种的培育奠定了基础。In this paper,a cDNA fragment of 450 bp which located on the 3′ terminal of polyphenol oxidase（PPO） gene was amplified from Yali pear by using RT-PCR method,then the antisense expression vector was constructed by inserting the fragment of Yali pear PPO gene between the CaMV promoter and NOS terminator of the expression vector pBI121 in reverse orientation.After that,with agrobacterium-mediated method,the PPO antisense gene was transformed into Yali plants.Northern blot analysis and Enzyme activity assay showed that the PPO activities in the transgenic Yali plants were significantly decreased compared with the non-transformed Yali plants.This will lay a good foundation for breeding new varieties of pears which resistant to browning in the future.

关 键 词：鸭梨 多酚氧化酶 反义表达载体 遗传转化 

分 类 号：S661.2[农业科学—果树学]