摘要
【目的】将抗逆相关转录因子ABP9基因导入大豆(Glycine max L.)基因组,并对转化系统条件进行探索,为提高大豆遗传转化效率提供依据。【方法】以农杆菌介导的半种法大豆遗传转化系统为基础,对目的基因进行转化;以健康外植体获得率、抗性丛生芽获得率和抗性芽伸长比率为指标,对转化系统条件进行优化。【结果】共培养时保证充足的光照和氧气,外植体生长情况最好;在共培养或诱导丛生芽培养基中加入600mg·L-1的L-半胱氨酸可以使抗性丛生芽获得率由22.1%提高至32.6%;采用4种植物激素进行正交试验,诱导抗性芽伸长比率最高的激素配比为:6-BA为1.5mg·L-1,GA3为0.75mg·L-1,IAA为0.1mg·L-1,ZT为1.0mg·L-1。【结论】利用优化的方法进行遗传转化研究已获得转基因再生植株,经PCR和DNA测序等分子检测,证明目的基因ABP9已导入并整合到大豆基因组中;转化率为1.33%。
[Objective] The present study was to introduce the transcription factor ABP9 into soybean genome and optimize the conditions of the genetic transformation system. [ Method ] The foreign gene was transformed into half-seed explants of soybean via Agrobacterium. The half-seed method system of Agrobacterium-mediated soybean transformation was researched applying healthy explants rates, resistant tufty shoots and elongated shoots as targets. [ Result ] Explants of soybean grow well in environments with abundant light and oxygen in cocultivation. Supplementation of L-cys （600 mg·L^-1） in culture media of cocultivation or shoot induction increased the rates of resistant tufty shoots from 22.1% to 32.6%. As the result of orthogonal experiment with four auxins, the concentrations made the highest proportion of shoots elongation were 6-BA 1.5 mg·L^-1, GA3 0.75 mg·L^-1, IAA 0.1 mg·L^-1 and ZT 1.0 mg·L^-1. [ Conclusion ] The regenerated planflets resistant to glufosinate were obtained. The PCR and DNA sequence results show that ABP9 gene was integrated into the genome of soybean. The rate of genetic transformation in cultivar Zhongdou was 1.33%.
出处
《中国农业科学》
CAS
CSCD
北大核心
2008年第7期1908-1916,共9页
Scientia Agricultura Sinica
基金
国家“863”快速反应课题
国家农作物基因资源与遗传改良重大科学工程开放课题
中国农业科学院作物科学研究所基金
国家自然科学基金重大课题(30490251)
