PLoS ONE (Jan 2012)

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,
  • Jin-Gui Chen

DOI
https://doi.org/10.1371/journal.pone.0044908
Journal volume & issue
Vol. 7, no. 9
p. e44908

Abstract

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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.