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Web End = Plant Cell Rep (2015) 34:12391251 DOI 10.1007/s00299-015-1783-z
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Web End = A highly efcient maize nucellus protoplast system for transient gene expression and studying programmed cell death-related processes
Jiang Chen1 Qiang Yi1 Qiaoheng Song3 Yong Gu1 Junjie Zhang2
Yufeng Hu3 Hanmei Liu2 Yinghong Liu1 Guowu Yu3 Yubi Huang1
Received: 13 February 2015 / Revised: 6 March 2015 / Accepted: 9 March 2015 / Published online: 20 March 2015 Springer-Verlag Berlin Heidelberg 2015
AbstractKey message Conditions for the isolation and transfection of maize nucellus protoplasts were established. We demonstrated its utilization for protein expression, localization, proteinprotein interaction, and the investigation of PCD-related processes.
Abstract 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 proteinprotein 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, b-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 efcient maize nucellus protoplast system for transient gene expression and investigation of PCD-related processes.
Keywords Protoplasts Maize nucellus PCD Transient
expression Cys-protease
Introduction
Plant protoplasts are cells surrounded only by the plasma membrane, but retain their viability. Because the protective cell wall has been removed, they can easily uptake bacteria, cell organelles, and nucleic acids from outside the cell (Davey and Cocking 1972; Ohyama et al. 1972). These characteristics make plant protoplasts a versatile cell-based experimental system...