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机构地区:[1]西北农林科技大学园艺学院/旱区作物逆境生物学国家重点实验室,陕西杨陵712100 [2]广西农业科学院蔬菜研究所
出 处:《长江蔬菜》2012年第18期10-14,共5页Journal of Changjiang Vegetables
基 金:国家科技支撑计划(2009BADB8B03);陕西省蔬菜产业体系;广西农业科学院基本科研业务专项项目(2011YZ17);广西农业科学院博士科研启动基金项目
摘 要:以06J28橙色大白菜子叶段为外植体,通过根癌农杆菌介导CMS7311-orf224基因,探讨了潮霉素、头孢霉素、预培养时间、农杆菌浓度、感菌时间和共培养时间等因素对橙色大白菜遗传转化的影响。试验结果表明,子叶段预培养2~3 d后,在OD600值为0.3~0.5的农杆菌EHA-105菌液中侵染5 min,再共培养2~3 d,在培养基中加入5 mg/LHyg(抗性筛选)和500 mg/L Cef(脱菌)可得到转化植株。试验初步建立了橙色大白菜遗传转化体系,为大白菜种质资源创新奠定了基础。In this study,several factors affecting the transformation rate were studied by Agrobacterium-mediated CMS7311-orf224 gene in 06J28 Chinese cabbage,and these factors included sensitivity of cotyledon segments to hygromycin(Hyg) and Cefotaxime(Cef),pre-culture days,concentrations of Agrobacterium tumefaciens,co-culture days and times of being infected on the optimum culture medium.The results showed that,resistant plants could be obtained with the conditions as follows: the cotyledon segments of 06J28 Chinese cabbage were pre-cultured for 2-3 days,and then infected with EHA-105(the OD600 value ranged from 0.3 to 0.5) for 5 minutes and were co-cultured for 2-3 days,lastly were transformed to screening-culture medium(adding 5 mg/L Hyg and 500 mg/L Cef).A higher efficient genetic transformation system was established for orange Chinese cabbage,which set up a basis for germplasm resources innovation of Chinese cabbage.
关 键 词:橙色大白菜 CMS7311-orf224基因 遗传转化
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