Velikova Violeta, Ghirardo Andrea, Vanzo Elisa, Merl Juliane, Hauck Stefanie M, Schnitzler Jörg-Peter
Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences , Acad. G. Bonchev Str. Bl. 21, 1113 Sofia, Bulgaria.
J Proteome Res. 2014 Apr 4;13(4):2005-18. doi: 10.1021/pr401124z. Epub 2014 Mar 20.
Biogenic isoprene (2-methyl-1,3-butadiene) improves the integrity and functionality of thylakoid membranes and scavenges reactive oxygen species (ROS) in plant tissue under stress conditions. On the basis of available physiological studies, we hypothesized that the suppression of isoprene production in the poplar plant by genetic engineering would cause changes in the chloroplast protein pattern, which in turn would compensate for changes in chloroplast functionality and overall plant performance under abiotic stress. To test this hypothesis, we used a stable isotope-coded protein-labeling technique in conjunction with polyacrylamide gel electrophoresis and liquid chromatography tandem mass spectrometry. We analyzed quantitative and qualitative changes in the chloroplast proteome of isoprene-emitting and non isoprene-emitting poplars. Here we demonstrate that suppression of isoprene synthase by RNA interference resulted in decreased levels of chloroplast proteins involved in photosynthesis and increased levels of histones, ribosomal proteins, and proteins related to metabolism. Overall, our results show that the absence of isoprene triggers a rearrangement of the chloroplast protein profile to minimize the negative stress effects resulting from the absence of isoprene. The present data strongly support the idea that isoprene improves/stabilizes thylakoid membrane structure and interferes with the production of ROS.
生物源异戊二烯(2-甲基-1,3-丁二烯)可改善类囊体膜的完整性和功能,并在胁迫条件下清除植物组织中的活性氧(ROS)。基于现有的生理学研究,我们推测通过基因工程抑制杨树中异戊二烯的产生会导致叶绿体蛋白质模式发生变化,进而补偿非生物胁迫下叶绿体功能和整体植物性能的变化。为了验证这一假设,我们将稳定同位素编码蛋白质标记技术与聚丙烯酰胺凝胶电泳和液相色谱串联质谱联用。我们分析了释放异戊二烯和不释放异戊二烯的杨树叶绿体蛋白质组的定量和定性变化。在此我们证明,通过RNA干扰抑制异戊二烯合酶会导致参与光合作用的叶绿体蛋白质水平降低,以及组蛋白、核糖体蛋白和与代谢相关的蛋白质水平升高。总体而言,我们的结果表明,异戊二烯的缺失会引发叶绿体蛋白质谱的重新排列,以尽量减少因异戊二烯缺失而产生的负面胁迫效应。目前的数据有力地支持了异戊二烯改善/稳定类囊体膜结构并干扰ROS产生的观点。