Highly Efficient Isolation of Populus Mesophyll Protoplasts and Its Application in Transient Expression Assays |
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Authors: | Jianjun Guo Jennifer L. Morrell-Falvey Jessy L. Labbé Wellington Muchero Udaya C. Kalluri Gerald A. Tuskan Jin-Gui Chen |
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Affiliation: | Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, United States of America.; University College Dublin, Ireland, |
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Abstract: | BackgroundPopulus 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 FindingsWe 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/SignificanceThis 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. |
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