农杆菌介导CSPs基因转化甘蓝型油菜转化体系的研究  

On System of Transforming CSPs Gene into Brassica napus Mediated by Agrobacterium

作　　者：谢琳[1] 杨娟[1] 代其林[1] 张新[1] 陈翠娜[1] 张思维[1] 王劲[2] 

机构地区：[1]西南科技大学极端环境生物资源利用实验室,四川绵阳621000 [2]中国农业科学院生物技术研究所,北京100084

出　　处：《绵阳师范学院学报》2011年第11期91-95,共5页Journal of Mianyang Teachers＇ College

基　　金：转基因生物新品种培育重大专项项目(2009ZX08009-091B);国家自然科学基金项目(30871555);教育部新世纪优秀人才支持计划(NCET-08-0940);四川省教育厅青年基金项目(09ZA034);西南科技大学博士研究基金项目(11zx7104)

摘　　要：以甘蓝型油菜下胚轴为外植体,通过农杆菌介导法,对其遗传转化条件进行了研究,建立了转冷激蛋白(CSPs)的遗传转化体系。结果表明,甘蓝型油菜下胚轴经预培养2d后,用浓度为OD600=0.4的农杆菌菌液侵染90s,再经共培养和愈伤诱导培养,外源基因转化到油菜下胚轴外植体的转化效率高。然后通过PCR检测转化后在筛选培养基上诱导成苗的油菜,初步确定CSPs基因已整合到油菜再生植株中。这为以后其他基因转化到甘蓝型油菜的研究奠定一定的基础。he system of genetic transformation into hypocotyl （as explant） of Brassica napus mediated by Agrobacterium was studied. The results demonstrate that there had high conversion efficiency for exogenous genes into the hypocotyl （explants） Brassica napus L after the hypocotyl was pre - cultured for 2d and then infected for 90s by Agrobacterium tumefaciens （OD600 = 0. g） with CSPs gene, and the infected hypocoty was co -cultured and cultured for induced - callus. The rape seedlings induced on the screening culture - medium were detected by PCR analysis, which proved that the CSPs gene had been integrated into the genome of regenerated rape. From above, it laid foundation for the research on other gene transformed into B. napus in future.

关 键 词：遗传转化 油菜下胚轴 农杆菌 冷激蛋白 

分 类 号：Q7[生物学—分子生物学]