Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, United States of America.
PLoS One. 2012;7(9):e44908. doi: 10.1371/journal.pone.0044908. Epub 2012 Sep 13.
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.
杨树是一种模式木本植物，也是木质纤维素生物燃料生产的很有前途的原料。然而，其漫长的生命周期阻碍了基因功能的快速特征描述。
方法/主要发现：我们优化了杨树叶片叶肉原生质体分离方案，并建立了杨树原生质体瞬时表达系统。我们证明了杨树原生质体能对激素刺激做出反应，并且一系列细胞器标记在杨树原生质体中正确定位。此外，我们表明，杨树原生质体瞬时表达系统适合研究蛋白质-蛋白质相互作用、基因激活和细胞信号事件。
结论/意义：本研究建立了一种从杨树叶片中高效分离原生质体的方法，并证明了使用杨树原生质体瞬时表达分析作为体内系统来描述基因和途径的有效性。
