河南农业大学学报2018，Vol.52Issue(1)：43-49,7.
‘蜂蜜罐’枣遗传转化条件的优化及Bt基因的导入
Optimization of genetic transformation system and transformation of Bt gene for epicotyl and cotyledon from seedlings of jujube'Fengmiguan'(Ziziphus jujuba Mill)
摘要
Abstract
The establishment of a high efficient and stable genetic transformation system is the basis for genetic improvement of plant germplasm by genetic engineering. In this study, the binary vector Pbi121 containing gus (β-glucuronidase) gene and the neomycin phosphotransferase Ⅱ (NPTⅡ) gene was introduced into jujube 'Fengmiguan' (Ziziphus jujuba Mill) by Agrobacterium-mediated transformation using epicotyls and cotyledons as explants. The effects of different bacterial concentra-tions,time of infection and co-cultivation,the concentration of acetosyringone (AS) and the infection pattern on the transient expression rate of guswere investigated. Bt (Bacillus thuringiensis) gene was also introduced into jujube 'Fengmiguan' by using the optimized genetic transformation system. The results showed that the most appropriate genetic transformation system by gus staining for the epicotyls and cotyledons was when bacterial culture reached OD600= 0.5 and infected for 10 min under vacu-um and then co-cultivated in a medium supplemented with 20 mg·L-1,10 mg·L-1of AS for 3 d, resepctively, the rate ofgus transient expression on epicotyls was significantly higher than cotyledons. Bt gene introduced into 'Fengmiguan' was carried out by using the optimized genetic transformation system and epicotyls and cotyledons were used as explants. After co-cultivation in a selective medium, the resistant shoots with the height of 1 ~1.5 cm were analyzed by PCR. Total 24 transgenic shoots were obtained and elongated from epicotyls and cotyledons. The transgenic plantlets with Bt gene were eventually obtained. Thus would provide a possibility for creating insect-resistant germplasm.关键词
'蜂蜜罐'枣/上胚轴/子叶/gus瞬时表达率/Bt基因Key words
'Fengmiguan'/epicotyl/cotyledon/the rate of gustransient expression/Bt gene分类
农业科技引用本文复制引用
张燕征,张梦洋,李继东,谭彬,王腾飞,郑先波,叶霞,冯建灿..‘蜂蜜罐’枣遗传转化条件的优化及Bt基因的导入[J].河南农业大学学报,2018,52(1):43-49,7.基金项目
河南省重大科技专项项目(151100110900) （151100110900）
河南省现代农业技术体系建设专项资金资助项目(S2014-11-G02) （S2014-11-G02）
