A highly efficient maize nucellus protoplast system for transient gene expression and studying programmed cell death-related processes
A highly efficient maize nucellus protoplast system for transient gene expression and studying programmed cell death-related processes
复制标题
用于瞬时基因表达和研究程序性细胞死亡相关过程的高效玉米珠心原生质体系统
DOI：
10.1007/s00299-015-1783-z
复制
发表时间：
2015-07-01
影响因子：
6.2
通讯作者：
Huang, Yubi
中科院分区：
文献类型：
--
作者：
Chen, Jiang;Yi, Qiang;Huang, Yubi
建立了玉米胞心原生质体的分离和转染条件。我们展示了它在蛋白质表达、定位、蛋白质相互作用和研究pcd相关过程中的应用。植物原生质体是一种重要的多功能细胞系统，广泛应用于基因表征和多种信号通路的分析。细胞程序性死亡（PCD）发生在植物从胚胎发育到受精的整个生命过程中。玉米胚囊发育过程中，珠心发生典型的PCD。胞心原生质体在pcd相关过程的研究中显示出潜在的应用前景。目前还没有关于胞心原生质体的分离和转染的研究报道。本研究建立了玉米核质原生质体的分离和转染条件。玉米原生质体系统可用于蛋白表达、定位和蛋白-蛋白相互作用。我们应用这个系统来调查与pcd相关的过程。实时荧光定量PCR分析表明，MADS29在玉米核质原生质体中的瞬时表达提高了cys -蛋白酶基因的转录水平。此外，β -葡萄糖醛酸酶和荧光素酶活性测定表明，MADS29可以增强cys -蛋白酶基因的启动子活性。因此，我们证明了一个高效的玉米核原生质体系统在瞬态基因表达和研究pcd相关过程中的潜力。
Conditions for the isolation and transfection of maize nucellus protoplasts were established. We demonstrated its utilization for protein expression, localization, protein-protein interaction, and the investigation of PCD-related processes.Plant protoplasts are an important and versatile cell system that is widely used in the analysis of gene characterization and diverse signaling pathways. Programmed cell death (PCD) occurs throughout the life of plants from embryogenesis to fertilization. The maize nucellus undergoes typical PCD during development of the embryo sac. The nucellus protoplast shows potential for use in research of PCD-related processes. No studies have reported previously the isolation and transfection of nucellus protoplasts. In this study, conditions for the isolation and transfection of maize nucellus protoplasts were established. The maize protoplast system can be used for protein expression, localization, and protein-protein interaction. We applied this system to investigate PCD-related processes. Quantitative real-time PCR analysis revealed that transient expression of MADS29 in the maize nucellus protoplast increases Cys-protease gene transcript level. In addition, beta-glucuronidase and luciferase activity assays showed that MADS29 could enhance the promoter activities of the Cys-protease gene. Thus, we demonstrated the potential of a highly efficient maize nucellus protoplast system for transient gene expression and investigation of PCD-related processes.