近日,北京林业大学生物科学与技术学院陈少良实验室利用“非损伤微测技术”取得的科研成果在《Plant, Cell & Environment》(2008 IF 4.666)杂志发表。这项成果将非损伤微测技术与激光共聚焦、X射线微量分析等技术相结合,研究了胡杨细胞在盐胁迫下与K+/Na+动态平衡调控相关的信号通路,发现质膜H+偶联转运体(PM H+-coupled transport system)可以激发H2O2及胞质Ca2+信号,进而通过调节K+通道及Na+/H+反向转运体调控K+/Na+动态平衡。非损伤微测技术在本研究中是获得离子跨膜流动信息的必不可少的工具。
陈少良实验室在一年多时间里利用非损伤微测技术已经连续发表三篇高水平的研究论文,达到甚至超过国外相同领域的科研水平,跻身世界一流实验室行列。该实验室已经积累了应用非损伤微测技术的丰富经验,培养出以本文第一作者孙健(也是前两篇研究论文的第一作者)为代表的一批熟练掌握非损伤微测技术的优秀学子,相信不久还会有大量的优质成果诞生。
ABSTRACT:
Using confocal microscopy, X-ray microanalysis and the scanning ion-selective electrode technique, we investigated the signalling of H2O2, cytosolic Ca2+ ([Ca2+]cyt) and the
PM H+-coupled transport system in K+/Na+ homeostasis control in NaCl-stressed calluses of Populus euphratica.An obvious Na+ /H+ antiport was seen in salinized cells;however, NaCl stress caused a net K+ efflux, because of the salt-induced membrane depolarization. H2O2 levels, regulated upwards by salinity, contributed to ionic homeostasis,because H2O2 restrictions by DPI or DMTU caused enhanced K+ efflux and decreased Na+ /H+ antiport activity. NaCl induced a net Ca2+ influx and a subsequent rise of [Ca2+]cyt, which is involved in H2O2-mediated K+/Na+ homeostasis in salinized P. euphratica cells. When callus cells were pretreated with inhibitors of the Na+ /H+ antiport system, the NaCl-induced elevation of H2O2 and [Ca2+]cyt was correspondingly restricted, leading to a greater K+ efflux and a more pronounced reduction in Na+ /H+ antiport activity. Results suggest that the PM H+-coupled transport system mediates H+ translocation and triggers the stress signalling of H2O2 and Ca2+, which results in a K+/Na+ homeostasis via mediations of K+ channels and the Na+ /H+ antiport system in the PM of NaCl-stressed cells. Accordingly, a salt stress signalling pathway of P. euphratica cells is proposed.