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农杆菌介导法将硝酸还原酶基因转入大白菜的研究
引用本文:张艳萍,陈玉梁,张正英,肖兴国.农杆菌介导法将硝酸还原酶基因转入大白菜的研究[J].安徽农业科学,2009,37(30):14597-14599.
作者姓名:张艳萍  陈玉梁  张正英  肖兴国
作者单位:甘肃省农业科学院生物技术研究所,甘肃,兰州,730070;中国农业大学,北京,100080
基金项目:甘肃省科学事业费项目 
摘    要:目的]建立大白菜的硝酸还原酶基因(NR)转化体系。方法]以大白菜北京小杂印为材料,通过农杆菌LBA4404介导,将硝酸还原酶基因(NR)转入大白菜,对获得的转NR基因苗进行PCR检测、Southern杂交检测以及硝酸盐含量测定。结果]试验初步建立了一套大白菜的转化体系,即以预培养2d苗龄4d的大白菜不带柄半子叶为外植体,在OD600值nm约为0.6的农杆菌工程菌液中浸染8min,然后在pH值为5.8且不加抗生素的分化培养基中将子叶与菌共培养2d,其后经过诱导、抗生素筛选、生根生长等过程,获得抗性植株,转化频率为6.48%。对抗性植株采用分级筛选,获得8株转NR基因苗。结论]试验证实,NR基因已整合进大白菜的基因组中并得到表达,植株体内的硝酸盐含量均低于对照。

关 键 词:大白菜  农杆菌  硝酸还原酶基因(NR)  硝酸盐含量

Study on Transformation of NR Gene into Chinese Cabbage by Agrobacterium Mediating Method
ZHANG Yan-ping et al.Study on Transformation of NR Gene into Chinese Cabbage by Agrobacterium Mediating Method[J].Journal of Anhui Agricultural Sciences,2009,37(30):14597-14599.
Authors:ZHANG Yan-ping
Institution:ZHANG Yan-ping et al (Institute of Biotechnology,Gansu Academy of Agricultural Science,Lanzhou,Gansu 730070)
Abstract:Objective] The purpose was to establish the nitrate reductase gene(NR) transformation system for Chinese cabbage.Method] With Chinese cabbage of Beijing Xiaoza 60 as material,the NR was transferred into Chinese cabbage through Agrobacterium LBA4404 mediating,the transgenic seedlings obtained were conducted for PCR detection and Southern blot as well as nitrate content determination.Result] A transformation system of Chinese cabbage was founded preliminarily in the test:with half cotyledon,without petiole...
Keywords:Chinese cabbage  Agrobacterium  Nitrate reductase(NR) gene  Nitrate content  
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