Highly Efficient Isolation of Populus Mesophyll Protoplasts and Its Application in Transient Expression Assays
Jianjun Guo, Jennifer L. Morrell-Falvey, Jessy L. Labbé, Wellington Muchero, Udaya C. Kalluri, Gerald A. Tuskan, and Jin-Gui Chen
20 January 2013, PLoS ONE 7(9): e44908; 10.1371/journal.pone.004908
Background: Populus is a model woody plant and a promising feedstock for lignocellulosic biofuel production. However, its lengthy life cycle impedes rapid characterization of gene function.
Methodology/Principal Findings: We optimized a Populus leaf mesophyll protoplast isolation protocol and established a Populus protoplast transient expression system. We demonstrated that Populus protoplasts are able to respond to hormonal stimuli and that a series of organelle markers are correctly localized in the Populus protoplasts. Furthermore, we showed that the Populus protoplast transient expression system is suitable for studying protein-protein interaction, gene activation, and cellular signaling events.
Conclusions/Significance: This study established a method for efficient isolation of protoplasts from Populus leaf and demonstrated the efficacy of using Populus protoplast transient expression assays as an in vivo system to characterize genes and pathways.
Guo J, Morrell-Falvey JL, Labbé JL, Muchero W, Kalluri UC, et al. (2012) Highly Efficient Isolation of Populus Mesophyll Protoplasts and Its Application in Transient Expression Assays. PLoS ONE 7(9): e44908. doi:10.1371/journal.pone.0044908