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萝卜遗传转化体系的建立
引用本文:李海萍,张鲁刚,胥宇建,范爱丽. 萝卜遗传转化体系的建立[J]. 中国瓜菜, 2011, 24(4)
作者姓名:李海萍  张鲁刚  胥宇建  范爱丽
作者单位:西北农林科技大学园艺学院,陕西杨凌,712100
摘    要:系统研究了影响根癌农杆菌介导的萝卜(Raphanus sativus L.)遗传转化的若干因素,建立了萝卜遗传转化体系:即切取萝卜带柄子叶外植体,先进行2 d预培养,用OD600为0.3~0.5的EHA105农杆菌菌液侵染5~7 min后,再进行5 d共培养,然后将外植体转接到MS+6-BA 6 mg.L-1+NAA 0.05 mg.L-1+Cef 500 mg.L-1的培养基上进行7 d延迟筛选,再将外植体转接到MS+6-BA 6 mg.L-1+NAA 0.05 mg.L-1+Cef 500 mg.L-1+Hyg 5 mg.L-1的培养基上进行10 d的抗性筛选。结果共获得126个抗性芽,其中8个抗性芽经PCR检测扩增出目的基因的启动子BcA9,约850bp,阳性率为6.3%,初步验证目的基因已经转入萝卜再生芽中。

关 键 词:萝卜  遗传转化  细胞质雄性不育基因

Establishment of Genetic Transformation System in Radish
LI Hai-ping,ZHANG Lu-gang,XU Yu-jian,FAN Ai-li. Establishment of Genetic Transformation System in Radish[J]. China Cucurbits And Vegetables, 2011, 24(4)
Authors:LI Hai-ping  ZHANG Lu-gang  XU Yu-jian  FAN Ai-li
Affiliation:LI Hai-ping,ZHANG Lu-gang,XU Yu-jian,FAN Ai-li(College of Horticulture,Northwest Agriculture and Forestry University,Yangling,Shaanxi,712100,China)
Abstract:The factors for Agrobacterium tumefaciens-mediated genetic transformation in Radish(Raphanus sativus L.)were studied.A radish genetic transformation system is established.Cotyledons with petiole were cultured in MS + 6-BA 6 mg·L-1 + NAA 0.05 mg·L-1 for 2 days as pre-culture.They were then infected for 5-7 min by the EHA105 Agrobacterium at OD600 = 0.3-0.5,and co-cultured in same medium for 5 days.The explants were then transferred to MS + 6-BA 6 mg·L-1 + NAA 0.05 mg·L-1 + 500 mg·L-1 Cef medium for 7 days for pre-screening.The explants were later cultured no MS + 6-BA 6 mg·L-1 + NAA 0.05 mg·L-1 + 500 mg·L-1 Cef + 5 mg·L-1 Hyg medium for 10 days for selecting transgenic events.We got 126 resistant shoots,and 8 of them were amplified the target gene BcA9 with size about 850 bp.The preliminary validation showed that the target gene has been transferred into radish.
Keywords:Radish  Genetic transformation  Cytoplasmic male sterility gene  
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