共培养的水分状态对农杆菌转化玉米的影响
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摘要
农杆菌与受体的共培养条件,对农杆菌介导的遗传转化有着重要的作用,为了提高玉米芽生长点的遗传转化效率,以郑58玉米自交系的种子为试验材料,用EGFP(增强型绿色荧光蛋白)基因作为标记基因,通过控制农杆菌共培养时的水分提供方式,研究共培养时的水分条件对农杆菌转化玉米芽生长点的影响。结果表明,将转化的玉米种子放到加有2层滤纸的培养皿中,每培养皿放置10～15粒种子,加1.5 mL无菌水,放在25℃暗培养3～4 d,在不影响转化效果的基础上有利于转化后发芽,可获得较多的转化材料。
The co-culture condition of agrobacterium and its receptor plays an important role in the agrobacterium-mediaed genetic transformation. The purpose of this study is to improve the genetic transformation efficiency of maize bud growth point. Using the seed of maize inbred line Zheng 58 as the experimental material, EGFP(enhanced green fluorescent protein) gene as reporter, and by controlling the water supply in agrobacterium co-culture, the effect of water condition on the growth point of maize germinating was studied. The results showed that when the transformed maize seeds was cultured in petri dishes with 2 layers of filter papers and 10 ～15 seeds and 1.5 mL sterile water for 3 ～4 d at 25 ℃ in dark, it was beneficial to germinate after transformation without affecting the transformation effect, and took more transformants.
The co-culture condition of agrobacterium and its receptor plays an important role in the agrobacterium-mediaed genetic transformation. The purpose of this study is to improve the genetic transformation efficiency of maize bud growth point. Using the seed of maize inbred line Zheng 58 as the experimental material, EGFP(enhanced green fluorescent protein) gene as reporter, and by controlling the water supply in agrobacterium co-culture, the effect of water condition on the growth point of maize germinating was studied. The results showed that when the transformed maize seeds was cultured in petri dishes with 2 layers of filter papers and 10 ～15 seeds and 1.5 mL sterile water for 3 ～4 d at 25 ℃ in dark, it was beneficial to germinate after transformation without affecting the transformation effect, and took more transformants.
引文
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[5]邹湘辉,庄东红,胡忠,等.负压和超声波处理对农杆菌介导的花生遗传转化效率的影响[J].中国油料作物学报, 2004,26(1):12-16.
[8]白云凤,王国英,苟明月,等.农杆菌介导玉米转基因的影响因素研究[J].中国生态农业学报,2006,14(3):173-175.
[6]王关林,方宏筠.植物基因工程[M].北京:科学出版社,2002:734-735.
[9]段学清,郑莉茹,刘文波,等. GFP绿色荧光法辅助筛选玉米转基因植株[J].玉米科学2017,25(2):31-38.
[10]张丽君,刘龙龙,张建珍,等.农杆菌介导创伤胚转化的应用[J].安徽农业科学, 2015, 43(3):25-28.
[2]张洁,周岩.根癌农杆菌转化单子叶植物的研究进展与对策[J].生物技术通报, 2013(5):7-14.
[3]沈平,章秋艳,林友华,等.推进我国转基因玉米产业化的思考[J].中国生物工程杂志, 2016, 36(4):24-29.
[4]袁鹰,李启云,郝文媛,等.农杆菌介导的玉米遗传转化影响因子的研究[J].分子植物育种,2006,4(2):228-232.
[7]李晓丽,王建军,崔瑞洁,等.大豆铁蛋白基因GmFerritin在玉米中的遗传转化[J].安徽农业大学学报, 2012, 39(2):263-268.
[5]邹湘辉,庄东红,胡忠,等.负压和超声波处理对农杆菌介导的花生遗传转化效率的影响[J].中国油料作物学报, 2004,26(1):12-16.
[8]白云凤,王国英,苟明月,等.农杆菌介导玉米转基因的影响因素研究[J].中国生态农业学报,2006,14(3):173-175.
[6]王关林,方宏筠.植物基因工程[M].北京:科学出版社,2002:734-735.
[9]段学清,郑莉茹,刘文波,等. GFP绿色荧光法辅助筛选玉米转基因植株[J].玉米科学2017,25(2):31-38.
[10]张丽君,刘龙龙,张建珍,等.农杆菌介导创伤胚转化的应用[J].安徽农业科学, 2015, 43(3):25-28.