CYcD2基因转入丽格海棠研究  

Preliminary Study on Transferring CYcD2 Gene into Begonia×hiemalis

作　　者：徐美隆[1] 张怀渝[2] 

机构地区：[1]种苗生物工程国家重点实验室,宁夏银川750004 [2]四川农业大学原子能农业应用研究室

出　　处：《安徽农业科学》2008年第19期8011-8013,共3页Journal of Anhui Agricultural Sciences

摘　　要：[目的]改变丽格海棠的开花时间,为丽格海棠分子育种奠定基础。[方法]以丽格海棠叶片为外植体,利用液氮直接转化法将pBI121-CYcD2转入根癌农杆菌EHA52中,再通过农杆菌介导法将CYcD2基因导入丽格海棠中。[结果]选择100 mg/L的卡那霉素作为转化外植体的起始选择压力。将经农杆菌共培养及过渡培养后的材料转入筛选培养基中,60 d后获得15个不定芽,再在生根培养基中培养20 d后,全部生根。PCR检测发现,15株卡那霉素抗性植株中有6株表现出与阳性对照相同的特异性条带,剩余植株为假转化体。Southern杂交分析表明6个转化植株中有5个出现了杂交信号,即为转基因植株,仅有1个为假阳性。[结论]该研究已成功地将外源基因CYcD2转入丽格海棠受体中。[Objective] The study aimed to change the flowering time of Begonia×hiemalis and lay a foundation for its molecular breeding. [Method] With Begonia×hiemalis leaves as explants,pBI121-CYcD2 was transferred into agrobacterium tumefacien EHA52 by direct liquid nitrogen transformation,and then CYcD2 gene was mediated into Begonia×hiemalis by agrobacterium-mediated method.[Result] 100 mg/L kanamycin was selected as initial selection pressure of transformed explants.The materials co-cultured with agrobacterium and cultured transitionally were transferred to screening medium and 15 adventitious buds were obtained after 60 d and all of them rooted after they were cultured in rooting medium for 20 d.It was found through PCR detection that the 6 of 15 plants with resistance to kanamycin showed specific bands identical to that of positive CK and the other plants were false transformants.Southern hybridization analysis showed the 5 of 6 transformed plants showed hybridization signals and were transgenic plants and only 1 was false positive plant.[Conclusion] The exogenous gene CYcD2 was transferred into Begonia×hiemalis receptor succcessfully in this study.

关 键 词：丽格海棠 CYcD2基因 遗传转化 农杆菌介导法 

分 类 号：S336[农业科学—作物遗传育种]