保卫细胞微管骨架参与蛋白丝氨酸/苏氨酸磷酸化调节的气孔运动

doi:10.3864/j.issn.0578-1752.2012.21.003

摘要/Abstract

摘要： 【目的】探讨在气孔运动的信息传递通路中，保卫细胞微管骨架与蛋白丝氨酸/苏氨酸磷酸化两种因素之间是否存在相互作用，进而深入了解气孔运动机理。【方法】以拟南芥野生型及GFP-α-tubulin-6植株为材料，利用药理学试验、激光共聚焦扫描显微镜观察、免疫印迹等细胞学及生物化学方法研究丝氨酸/苏氨酸蛋白磷酸化对保卫细胞气孔运动及微管骨架的影响。【结果】丝氨酸/苏氨酸蛋白激酶抑制剂星型孢菌素（staurosporine，STS）能促进光照下气孔开放，微管特异性抑制剂长春花碱（vinblastine）和微管稳定剂紫杉醇（taxol）分别减弱和增强其作用；1/2A型磷酸酶抑制剂冈田酸（okadaic acid，OA）、花萼海绵诱癌素A（calyculin A，CalA）抑制光照诱导的气孔开放，微管特异性药物长春花碱和紫杉醇又能分别增强和削弱该抑制作用。激光共聚焦扫描显微镜下观察，两种磷酸酶抑制剂OA和CalA处理后，正常开放气孔保卫细胞中整齐有序的辐射状微管数量显著减少，变为以交错网状及解聚态为主；蛋白激酶抑制剂STS处理则增加了辐射状微管的比例。微管特异性药物紫杉醇、长春花碱分别与以上3种抑制剂共同处理，同样能够在一定程度上改变其对保卫细胞微管骨架组织排布的影响。提取保卫细胞原生质体总蛋白进行免疫印迹检测，55 kD分子量处的微管蛋白发生明显的蛋白丝氨酸磷酸化。【结论】蛋白丝氨酸/苏氨酸磷酸化可以通过调节保卫细胞微管骨架的动态排布进行气孔运动的信息传递。

关键词: 微管骨架, 丝氨酸/苏氨酸蛋白磷酸化, 气孔运动

Abstract: 【Objective】 The interaction between microtubules (MTs) and protein serine/threonine phosphorylation in guard cells was investigated in order to provide a reliable evidence for better understanding of the mechanism of stomatal movment. 【Method】Pharmacological analysis, confocal laser scanning microscopy, and western blotting were used to explore the possible interaction of these two elements. 【Result】 In vivo stomatal aperture assays revealed that staurosporine (STS), one well-known specific inhibitor of serine/threonine protein kinase, enhanced the light-induced stomatal opening. While, okadaic acid (OA) and calyculin A (CalA), another two inhibitors of 1/2A phosphatase, showed the opposite effects under the same condition. In addition, it was found that these changes of stomatal movement could be reversed or became intensified to some extent when microtubule-specific drug, taxol or vinblastine was present during the treatments above. Furthermore, OA and CalA also had significant influences on the MT organization in the opening guard cells. In the presence of OA or CalA, the fine radial MT arrays tended to be broken down or rearranged into random network. And guard cells kept in STS showed increase in radial MT arrays and stomatal aperture. MT-targeted drugs, taxol and vinblastine, also altered the effects of three inhibitors (OA, CalA, STS) on MT organization partiallly in a similar manner. Immunoblot detection of total extract from guard cells confirmed that tubulin-subunits were phosphorylated on serine. 【Conclusion】 The results suggest that reversible protein serine/threonine phosphorylation may regulate the MT dynamics of guard cells during stomatal movement.

Key words: microtubule, protein serine/threonine phosphorylation, stomatal movement

叶子, 黄聪聪, 于荣. 保卫细胞微管骨架参与蛋白丝氨酸/苏氨酸磷酸化调节的气孔运动[J]. 中国农业科学, 2012, 45(21): 4351-4360.

YE Zi, HUANG Cong-Cong, YU Rong. Microtubules in Guard Cells Involve in Serine/Threonine Phosphorylation During Stomatal Movement[J]. Scientia Agricultura Sinica, 2012, 45(21): 4351-4360.

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