title: A highly efficient maize nucellus protoplast system for transient gene expression and studying programmed cell death-related processes.
abstract: 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, β-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.
authors: Key Laboratory of Biology and Genetic Improvement of Maize in Southwest Region, Maize Research Institute, Sichuan Agricultural University, 211 Huiming Road, Chengdu, 611130, China.
authors: Jiang
authors: J
authors: Chen
authors: Qiang
authors: Q
authors: Yi
authors: Qiaoheng
authors: Q
authors: Song
authors: Yong
authors: Y
authors: Gu
authors: Junjie
authors: J
authors: Zhang
authors: Yufeng
authors: Y
authors: Hu
authors: Hanmei
authors: H
authors: Liu
authors: Yinghong
authors: Y
authors: Liu
authors: Guowu
authors: G
authors: Yu
authors: Yubi
authors: Y
authors: Huang
mesh_terms: D017209:Apoptosis; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D009997:Osmotic Pressure; D010940:Plant Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified; D054817:Pollination; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D060066:Protein Interaction Maps; D021381:Protein Transport; D011523:Protoplasts / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection; D003313:Zea mays / Q000166:cytology* / Q000235:genetics*
chemical_list: D010940:Plant Proteins