农杆菌介导的河北杨遗传转化体系的建立  

Establishment of System for Genetic Transformation of Agrobacterium tumefaciens Mediated Transformation of Populus hopeiensis with Growth-promoting Gene

作　　者：王沛雅[1,2] 杨晖[1,2] 杨涛[1,2] 张军[1,2] 郭琪[1,2] 强维亚[3] 周剑平[2,4] 

机构地区：[1]甘肃省科学院生物研究所,兰州730000 [2]甘肃省工业微生物工程技术研究中心,兰州730000 [3]兰州大学生命科学学院,兰州730000 [4]甘肃省科学院,兰州730000

出　　处：《生物技术通报》2012年第3期141-147,共7页Biotechnology Bulletin

基　　金：甘肃省国际科技合作项目(0708WCGA140);甘肃省科学院应用技术研究与开发计划项目(2007YS-JK-05);甘肃省工业生物过程工程创新服务平台资助项目(094TTPA002)

摘　　要：以兼具生态和能源植物功能的木本模式植物——杨树(河北杨)为材料,研究了携带促生长基因(35S-DAS5)的根癌农杆菌载体介导的河北杨遗传转化若干因素对转化效果的影响。结果显示,较适宜的转化系统为预培养2-4 d,农杆菌菌液(OD600值为0.4)侵染20 min,共培养4 d,在含30 mg/L卡那霉素(Km)的培养基上诱导不定芽,生根培养基中Km的适宜浓度为10 mg/L。Several factors affecting transformation of Populus hopeiensis were studied by infecting with Agrobacterium tumefaciens GV3101,such as pre-culture time,adding AS,Agrobacterium concentration,infection time,co-culture time,concentration of kanamycin used for selection and adding time,of which the frequency of kanamycin resistance buds was used as measure standard.The superior transformation system was as following,pre-cultured 2-4 days and then infected 20 minutes in Agrobacterium diluted（OD600=0.4）,co-cultured 4 days,transgenic tissues and shoots were selected with 30 mg/L Km and rooting stage with 10 mg/L Km.

关 键 词：河北杨 根癌农杆菌 GV3101 35S-DAS5 遗传转化体系 

分 类 号：S792.11[农业科学—林木遗传育种]