《植物生理学报》 2016, 52(4): 575-579

根癌农杆菌介导GUS基因转化玉米芽尖生长点的研究

邢小龙¹, 常纪苹¹, 付忠军², 胡德升¹, 杨慧丽¹, 付小康¹, 钟立华¹, 胡彦民^1,*

¹河南农业大学农学院/河南粮食作物协同创新中心, 郑州450002; ²重庆市农业科学院, 重庆401329

通信作者：胡彦民；E-mail: huyanmin007@163.com

摘　要：

本试验以玉米(Zea mays)杂交种‘郑单958’的芽尖生长点作为农杆菌侵染受体, 以β-葡萄糖醛酸苷酶基因(GUS)作为转化基因, 吸水链霉菌草丁膦乙酰转移酶基因(bar)作为筛选标记基因, 通过GUS颜色反应, 分别从侵染液浓度、侵染时间和真空处理时间三个方面分析了影响GUS基因转化率的因素, 初步得出结论: 当侵染液浓度(OD₆₀₀)为0.6、侵染时间2 h、真空处理时间为10 min时, 选用杂交种‘郑单958’作为受体材料将会达到较高的遗传转化率。

关键词：玉米; 芽尖生长点; GUS基因; 根癌农杆菌; 遗传转化

收稿：2016-02-16   修定：2016-03-14

资助：国家自然科学基金(31071431)。

Study on Agrobacterium tumefaciens–mediated transformation of GUS gene into maize shoot apical growing point

XING Xiao-Long¹, CHANG Ji-Ping¹, FU Zhong-Jun², HU De-Sheng¹, YANG Hui-Li¹, FU Xiao-Kang¹, ZHONG Li-Hua¹, HU Yan-Min^1,*

¹College of Agronomy, Henan Agricultural University / Collaborative Innovation Center of Henan Grain Crops, Zhengzhou 450002, China; ²Chongqing Academy of Agricultural Sciences, Chongqing 401329, China

Corresponding author: HU Yan-Min; E-mail: huyanmin007@163.com

Abstract:

Using shoot apical growing point of maize hybrid ‘zhengdan958’ seeds as explants, several factors affecting the Agrobacterium tumefaciens–mediated genetic transformation were studied with β-glucuronidase (GUS) as transformation gene and Streptomyces hygroscopicus phosphinothricin acetyltransferase (bar) as screening gene. The results indicate that good transformation efficiency can be obtained when the concentration of bacterial OD₆₀₀ is 0.6, the co-cultivation time is 2 h, the vacuum infiltration time is 10 min and the receptor is ‘zhengdan958’.

Key words: maize; shoot apical growing point; GUS gene; Agrobacterium tumefaciens; genetic transformation