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PLoS One. 2012;7(9):e44908. doi: 10.1371/journal.pone.0044908. Epub 2012 Sep 13.

Highly efficient isolation of Populus mesophyll protoplasts and its application in transient expression assays.

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  • 1Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, United States of America.



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.


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.


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.

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