农杆菌介导韭菜遗传转化相关因素的研究

生物技术通报 ›› 2014, Vol. 0 ›› Issue (1): 125-131.

农杆菌介导韭菜遗传转化相关因素的研究

常婧¹,高行英¹,李小东²,侯雷平¹,李梅兰¹

1. 山西农业大学园艺学院，太谷 030801
2. 方兴现代农业有限公司，长治 046600

收稿日期:2013-09-06 出版日期:2014-01-23 发布日期:2014-01-23
作者简介:常婧，女，硕士研究生，研究方向: 蔬菜育种及生物技术；E-mail: 412567956@qq.com
基金资助:
山西省农业科技攻关项目(20090311022，0110311015-1)，山西省人事厅人才引进项目

Research on the Factors Related to Agrobacterium-mediated Genetic Transformation of the Chinese Chive

Chang Jing¹, Gao Xingying¹, Li Xiaodong², Hou Leiping¹, Li Meilan¹

1. College of Horticulture，Shanxi Agriculture University，Taigu 030801;
2. Fangxing Modern Agriculture Co. Ltd, Changzhi 046600

Received:2013-09-06 Published:2014-01-23 Online:2014-01-23

摘要/Abstract

摘要： 以“汉中冬韭”韭菜品种为试验材料，用含有pCAMBIA3301 质粒的根癌农杆菌菌株EHA105 对影响韭菜遗传转化#br#效率的多种因素进行研究。结果表明，诱导40 d 的愈伤组织，GUS 基因瞬时表达率达到93%，且最适宜于不定芽的分化；当乙酰#br#丁香酮(AS)的浓度为100 μmol/L 时，愈伤的GUS 表达率达到91%，植株再生率为7.9%，AS 浓度增加时其值也不会增加；菌液#br#OD600 值为0.6 侵染10 min 时，与其他组合相比，外植体受伤程度小，GUS 表达率及再生率最高；侵染后的愈伤共培养3 d 后，农#br#杆菌生长较少，GUS 表达率为91.1%，而再生率达到7.2%，为最佳的共培养时间。通过试验得到韭菜遗传转化因素的最佳条件，#br#为今后的遗传转化提供一些参考。

关键词: 韭菜, 农杆菌, GUS 染色, 遗传转化

Abstract: Using “Hanzhong Dongjiu” variety and the Agrobacterium EHA105 containing pCAMBIA3301 plasmid as the material, it studied the factors which could influence transformation efficiency of Chinese chive. The results showed that transient expression rate of GUS gene reached 93% with the callus cultured for 40 d as the explant and the adventitious buds differentiated well. When 100 μmol/L acetosyringone was added to the miedium, GUS expression rate of the callus reached 91% and the plant regeneration rate was 7.9%, but did not increase with increasing of the AS concentration. Compared with other combinations, the calli were soaked in Agrobacterium(OD600 ≈ 0.6)for 10 min, the injury degree of explants was slight and the GUS expression rate and regeneration rate were the highest. When the calli were co-cultured for 3 days after infection, the bacterium grew less than other period, GUS expression rate and the regeneration rate were 91.1% and 7.2% respectively. Through the experiment, the best conditions of genetic transformation factors for chinese chive were obtained.

Key words: Chinese chive, Agrobacterium, GUS staining, Genetic transformation

常婧,高行英,李小东,侯雷平,李梅兰. 农杆菌介导韭菜遗传转化相关因素的研究[J]. 生物技术通报, 2014, 0(1): 125-131.

Chang Jing, Gao Xingying, Li Xiaodong, Hou Leiping, Li Meilan. Research on the Factors Related to Agrobacterium-mediated Genetic Transformation of the Chinese Chive[J]. Biotechnology Bulletin, 2014, 0(1): 125-131.

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