潮霉素抗性油菜遗传转化体系优化  

Optimization of genetic transformation system based on hygromycin selection in rapeseed

作　　者：刘婧琳 范世航 华玮[1,2] 李俊 LIU Jing-lin;FAN Shi-hang;HUA Wei;LI Jun(Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences,Key Laboratory of Biology and Ge-netic Improvement of Oil Crops,Ministry of Agriculture and Rural Affairs,Wuhan 430062,China;Hubei Hongshan Laboratory,Wuhan 430070,China)

机构地区：[1]中国农业科学院油料作物研究所/农业农村部油料作物生物学与遗传育种重点实验室,湖北武汉430062 [2]湖北洪山实验室,湖北武汉430070

出　　处：《中国油料作物学报》2023年第3期518-523,共6页Chinese Journal of Oil Crop Sciences

基　　金：中国农业科学院科技创新工程重大科研任务(CAAS-ZDRW202105);国家自然科学基金(32072096)。

摘　　要：油菜分子育种有赖于高效的遗传转化技术。为提高甘蓝型油菜的遗传转化效率,采用油菜品种中双6号为材料,以下胚轴为外植体,通过农杆菌介导遗传转化,通过改变筛选培养基状态,增加液体培养基筛选处理的方式,对以潮霉素为筛选剂的油菜遗传转化体系进行了优化。在传统的固体筛选培养基培养21 d后,以液体筛选培养基形式对愈伤组织进行3 d再筛选培养。与传统方法相比,优化后再生苗形成提前约一周,出苗率提高6.94%,转基因苗阳性率提高15%,总转化效率提高6%。综上,该优化后的方法可为油菜基因功能解析与转基因育种研究提供有效技术支撑。Molecular breeding of rapeseed depends mostly on genetic transformation.To optimize the pro-cess of Agrobacterium-mediated genetic transformation system(hypocotyl as explant)based on hygromycin se-lection,we utilized cultivar Zhongshuang 6 as material by changing the state of the selection medium and adding liquid medium for selection.After culturing in traditional solid selection medium for 21 days,callus was re-se-lected for 3 days in the liquid selection medium.Compared to traditional methods,the regeneration seedlings formed about one week earlier with~6.94%increment of formation rate.In addition,the positive rate of transgen-ic plants increased by about 15%and the total transgenic efficiency increased by about 6%.In conclusion,the op-timized method reported here provides effective technical support for gene function analysis and transgenic breed-ing in rapeseed.

关 键 词：甘蓝型油菜 遗传转化 潮霉素 下胚轴 农杆菌介导法 

分 类 号：S565.4[农业科学—作物学]