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当前位置: 首页 > 文章 > 根癌农杆菌介导蓝猪耳转化的影响因素研究 亚热带植物科学 2008,37 (2) 17-20
Position: Home > Articles > Optimization of Agrobacterium-mediated Transformation System for Torenia fournieri Subtropical Plant Science 2008,37 (2) 17-20

根癌农杆菌介导蓝猪耳转化的影响因素研究

作  者：
龙海涛;李洪清;李玲
单  位：
华南师范大学生命科学学院广东省植物发育生物工程重点实验室
关键词：
蓝猪耳;根癌农杆菌;转化;影响因子
摘  要：
研究了根癌农杆菌介导蓝猪耳转化的影响因子。结果表明,以蓝色花类型蓝猪耳5～6周的叶片为外植体,在OD600为0.5～0.6的活化菌液中浸染5～10min,暗培养4d后,在愈伤组织诱导培养基(MS+BA1.0mg/L+2,4-D0.1mg/L)上生长14d,获得抗性愈伤组织;经芽诱导培养基(1/2MS+BA1.0mg/L+NAA0.1mg/L)培养28d得到抗性芽;生根培养基(1/2MS)上培养14d得到抗性植株。经PCR检测证实外源基因已整合到蓝猪耳基因组中,转化率达13%～14%。Cef和Hyg浓度对转化影响较大,转化的不同阶段其适宜浓度不同。
译  名：
Optimization of Agrobacterium-mediated Transformation System for Torenia fournieri
作  者：
LONG Hai-tao,LI Hong-qing,LI Ling(Guangdong Key Laboratory of Biotechnology for Plant Development,College of Life Science,South China Normal University,Guangzhou 510631,Guangdong China)
关键词：
Torenia fournieri;Agrobacterium tumefacien;transformation;factor
摘  要：
An efficient and stable Agrobacterium-mediated transformation system for Torenia was established through investigating the factors influencing the transformation efficiency.Genetic transformation of Torenia was performed by using 5～6 week leaflet as explants.After incubation in Agrobacterium suspension of OD600 at 0.5～0.6 for 5～10min,the explants were co-cultivated on co-culture medium containing 200μmol/L acetosyringone for 4d under dark.The explants were then transferred to the calla induction agar medium MS + BA 1.0mg/L + 2,4-D 0.1mg/L,and cultivated for 14d to obtain the resistant calla.The resistant calla were transferred to the shoot induction agar medium 1/2MS + BA 1.0mg/L + NAA 0.1mg/L,and cultured for another 28d to obtain resistant shoots.The 0.5cm or taller resistant shoots were finally transferred to rooting agar medium,and the resistant plantlets were achieved after 14d cultivation.Hygromycin-resistant plants were confirmed to be positive transformants by PCR.The transformation frequency was up to 13%～14% overall.

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