摘要
为建立高效稳定的玉米叶片原生质体瞬时转化体系,以玉米品种农大108叶片为材料,用聚乙二醇(polyethylene glycol,PEG 4000)介导的瞬时转化法将报告基因增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)转化到玉米原生质体中,分析PEG 4000浓度、转化时间和质粒用量对其原生质体转化效率的影响。结果表明:当PEG 4000浓度为40%、转化时间为15 min、质粒用量为7.5μg时,玉米叶片原生质体转化效率最高,为76.98%。
The recalcitrant nature of maize necessitates the development of an efficient and stable transient transformation system.Using the leaves of the maize cultivar Nongda 108 as explants,this study developed a transformation system and optimized its efficiency.The reporter gene encoding the enhanced green fluorescent protein(EGFP)was transformed into maize protoplasts by using polyethylene glycol(PEG 4000)-mediated transient transformation method.We determined the effects of PEG 4000 concentration,transformation time,and plasmid dosage on the transformation efficiency of maize protoplasts.The results showed that the transformation system achieved the maximum transformation efficiency of 76.98%under the optimal conditions as follows:the PEG 4000 concentration at40%,the plasmid coverage of 7.5μg,and the time interval of 15 minutes.The optimal conditions constitute a cost-effective and efficient transient transformation system of maize leaf protoplasts,laying the foundation for maize functional genomics studies.
作者
赵小慧
郁凯
刘冲
钟明娟
贺亭亭
董静
王凯
邢锦城
ZHAO Xiao-hui;YU Kai;LIU Chong;ZHONG Ming-juan;HE Ting-ting;DONG Jing;WANG Kai;XING Jin-cheng(Jiangsu Coastal Area Institute of Agricultural Sciences,Yancheng 224002,China;Zhucheng Municipal Bureau of Agricultural and Rural Affairs,Zhucheng 262200,China)
出处
《大麦与谷类科学》
2022年第6期6-10,共5页
Barley and Cereal Sciences
基金
国家自然科学基金(31901856)
苏北科技专项(SZ-YC202163)。
关键词
玉米
原生质体
瞬时转化体系
体系优化
Maize
Protoplast
Transient transformation system
Optimization
