核农学报2011，Vol.25Issue(4)：665-672,8.
农杆菌介导的大豆成熟胚芽尖遗传转化体系的优化
OPTIMIZATION OF AGROBACTERIUM-MEDIATED TRANSFORMATION SYSTEMS OF SOYBEAN MATURE EMBRYONIC TIP
摘要
Abstract
Transcription factor gene GmGT-2A was introduced into soybean variety Zhechun 3 via Agrobacterium-mediated soybean mature embryonic tip transformation systems.The effects of three main factors on the transformation efficiency,including pre-culture time of embryonic tips,6-BA concentration in the co-culture medium and the infection time were studied.Single factor optimizing test showed that transformation efficiency was affected by all the three factors.The optimal levels for each factor was 1.5d for pre-culture time,10h for infection time and 8mg/L for 6-BA concentration in the co-culture medium.Results of further orthogonal test showed that these three factors could be ranked as infection time,pre-culture time,and 6-BA concentration according to their effectiveness on transformation efficiency.The optimal transformation system at the same levels for each factor as mentioned above resulted in a highest transformation efficiency of 5.67%.A preliminary test also showed that rooting medium without IBA addition was better than that with 0.2mg/L IBA in terms of root emerging time and roots qualities.The transformation events were confirmed using target gene-specific,selective marker gene-specific and construction-specific PCR analysis.关键词
大豆/根癌农杆菌/胚芽尖/遗传转化Key words
soybean/Agrobacterium tumefaciens/embryonic tip/transformation分类
农业科技引用本文复制引用
杨海英,朱丹华,胡小丽,杨清华,袁凤杰,董德坤..农杆菌介导的大豆成熟胚芽尖遗传转化体系的优化[J].核农学报,2011,25(4):665-672,8.基金项目
转基因生物新品种培育重大专项 （）
