农杆菌介导甘蔗愈伤组织遗传转化体系的优化

中国生物工程杂志

2013, Vol. 33

Issue (9): 66-72

技术与方法

农杆菌介导甘蔗愈伤组织遗传转化体系的优化

秦翠鲜^1,2, 陈忠良^1,2, 桂意云^1,2, 汪淼^1,2, 周建辉^2,3, 廖青^2,3, 李杨瑞^1,2, 黄东亮^1,2

1. 广西农业科学院甘蔗研究所广西甘蔗遗传改良重点实验室农业部广西甘蔗生物技术与遗传改良重点实验室南宁 530007;
2. 中国农业科学院甘蔗研究中心南宁 530007;
3. 广西农业科学院南宁 530007

Optimization of Genetic Transformation System of Sugarcane Callus Mediated by Agrobacterium

QIN Cui-xian^1,2, CHEN Zhong-liang^1,2, GUI Yi-yun^1,2, WANG Miao^1,2, ZHOU Jian-hui^2,3, LIAO Qing^2,3, LI Yang-rui^1,2, HUANG Dong-liang^1,2

1. Sugarcane Research Institute, Guangxi Academy of Agricultural Sciences, Guangxi Key Laboratory of Sugarcane Genetic Improvement, Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture, P.R.China, Nanning 530007, China;
2. Sugarcane Research Center, Chinese Academy of Agricultural Sciences, Nanning 530007, China;
3. Guangxi Academy of Agricultural Sciences, Nanning 530007, China

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摘要： 以甘蔗胚性愈伤组织为材料,对愈伤组织增殖、分化、幼苗生根的最适抗生素(Km)浓度进行筛选,并对影响农杆菌介导的甘蔗遗传转化因素:预培养时间、侵染时间和共培养时间等进行了研究。最后确立了甘蔗愈伤组织增殖、分化及幼苗生根三个阶段的最适抗生素(Km)浓度分别为300 mg/L,40 mg/L,20 mg/L。确定了农杆菌浸染的预培养时间、侵染时间和共培养时间分别为4d,30min,4d。PCR检测表明,61.6%的抗性植株可扩增出预期的片段,表明外源基因已经整合到甘蔗基因组中。为进一步的甘蔗基因功能验证及转基因研究提供良好的技术支撑。

关键词： 甘蔗; 农杆菌; 遗传转化

Abstract: To establish high-efficiency sugarcane transformation system, the optimal concentration of antibiotic (Km) was screened for subculture and differentiation of sugarcane calli, as well as seedling rooting, respectively. The factors that influenced sugarcane transformation, such as pre-culture time, infection time and co-culture time were also studied. The results showed that the optimal concentrations of antibiotic (Km) for subculture, differentiation and seedling rooting were 300 mg/L, 40 mg/L and 20 mg/L, respectively. The optimal pre-culture time, infection time and co-culture time were 4d, 30min and 4d, respectively. PCR detection showed that 61.6% of resistance plants were transformed plants. This work lays a solid foundation for further study on gene function and transgenic research of sugarcane.

Key words: Sugarcane Agrobacterium Genetic transformation

收稿日期: 2013-06-19 出版日期: 2013-09-25

ZTFLH:

Q943

基金资助: 国家自然科学基金(31160301);广西自然科学基金(2011GXNSFF018002、2012GXNSFDA053011);广西农业科学院基本科研业务专项(桂农科2012YZ11、桂农科2012JM06、桂农科2013YM41、桂农科2013YM09)资助项目

通讯作者: 李杨瑞liyr@gxaas.net;黄东亮huangdl@gxaas.net E-mail: liyr@gxaas.net;huangdl@gxaas.net

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引用本文:

秦翠鲜, 陈忠良, 桂意云, 汪淼, 周建辉, 廖青, 李杨瑞, 黄东亮. 农杆菌介导甘蔗愈伤组织遗传转化体系的优化[J]. 中国生物工程杂志, 2013, 33(9): 66-72.

QIN Cui-xian, CHEN Zhong-liang, GUI Yi-yun, WANG Miao, ZHOU Jian-hui, LIAO Qing, LI Yang-rui, HUANG Dong-liang,. Optimization of Genetic Transformation System of Sugarcane Callus Mediated by Agrobacterium. China Biotechnology, 2013, 33(9): 66-72.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I9/66

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