Up-regulation and interaction of the plasma membrane H(+)-ATPase and the 14-3-3 protein are involved in the regulation of citrate exudation from the broad bean (Vicia faba L.) under Al stress.

质膜 H （ ） - ATPase 和 14 - 3 - 3 蛋白的上调和相互作用参与了 Al 胁迫下蚕豆 （ 蚕豆 ） 柠檬酸盐渗出的调节。

• 影响指数: 5.437
• 发表时间: Sep 2013
• 来源期刊: Plant Physiol Biochem
• DOI: 10.1016/j.plaphy.2013.06.015
• PMID: 23860230
• 文章类型: Journal Article
• 中科院分区: Q1
• 方向: 生物
• 作者列表: Chen Q,Guo CL,Wang P,Chen XQ,Wu KH,Li KZ,Yu YX,Chen LM

关键词: 1,3-bis-[tris(hy- droxymethyl)-methylamino]-propane; 14-3-3 Protein; 5’-AMP; AD; Al; Al stress; BTP; Broad bean; Citrate exudation; DDT; FC; OA; PM; PMSF; Plasma membrane H(+)-ATPase; VHA2p; Vicia faba 14-3-3-like protein b; Vicia faba plasma membrane H(+)-ATPase; Vicia faba. L cv. Anhui Candou; Vicia faba. L cv. Yunnan Candou 8363; YD; adenosine 5′-monophosphate; aluminum; dithiothreitol; fusicoccin; organic acid; phenylmethylsulfonyl fluoride; phosphorylated VHA2 at Thr-951 in C terminus; plasma membrane; vf14-3-3b; vha2

Mesh: 14-3-3 Proteins / metabolism; Adaptation, Physiological / genetics; Adenosine Monophosphate / pharmacology; Aluminum / adverse effects metabolism; Cell Membrane / metabolism; Citric Acid / metabolism; Gene Expression Regulation, Plant; Genes, Plant; Glycosides / pharmacology; Phosphorylation; Plant Proteins / genetics metabolism; Proton-Translocating ATPases / genetics metabolism; Stress, Physiological / genetics; Transcription, Genetic / drug effects; Up-Regulation; Vicia faba / drug effects genetics metabolism

Abstract

Our previous study showed that citrate excretion coupled with a concomitant release of protons was involved in aluminum (Al) resistance in the broad bean. Furthermore, genes encoding plasma membrane (PM) H(+)-ATPase (vha2) and the 14-3-3 protein (vf14-3-3b) were up-regulated by Al in Al-resistant (YD) broad bean roots. In this study, the roles of PM H(+)-ATPase (E.C. 3.6.3.6) and the 14-3-3 protein in the regulation of citrate secretion were further investigated in Al-resistant (YD) and Al-sensitive (AD) broad bean cultivars under Al stress. The results showed that greater citrate exudation was positively correlated with higher activities of PM H(+)-ATPase in roots of YD than AD. Real-time RT-PCR analysis revealed that vha2 was clearly up-regulated by Al in YD but not in AD roots, whereas the transcription levels of vf14-3-3b were elevated in a time-dependent manner in both YD and AD roots. Immunoprecipitation and Western analysis suggested that phosphorylation and interaction with the vf14-3-3b protein of the VHA2 were enhanced in YD roots but not in AD roots with increasing Al treatment time. Fusicoccin or adenosine 5\'-monophosphate increased or decreased the interaction between the phosphorylated VHA2 and the vf14-3-3b protein, followed by an enhancement or reduction of the PM H(+)-ATPase activity and citrate exudation in both cultivars under Al stress conditions, respectively. Taken together, these results suggested that Al enhanced the expression and interaction of the PM H(+)-ATPase and the 14-3-3 protein, which thereby led to higher activity of the PM H(+)-ATPase and more citrate exudation from YD plants.

摘要

我们先前的研究表明，柠檬酸盐的排泄以及伴随的质子释放与蚕豆的铝（Al）抗性有关。此外，编码质膜（PM）H（）-ATPase（vha2）和14-3-3蛋白（vf14-3-3b）的基因在抗Al（YD）蚕豆根中被Al上调。在这项研究中,进一步研究了抗Al（YD）和Al敏感（AD）蚕豆品种中PMH（）-ATPase（E.C.3.3.6.3.6.6）和14-3-3蛋白在柠檬酸盐分泌调节中的作用在Al胁迫下。结果表明，与AD相比，YD根中的柠檬酸渗出量较大与PMH（）-ATPase活性较高呈正相关。实时RT-PCR分析显示，vha2在YD中明显被Al上调，而在AD根中未被Al上调，而vf14-3-3b的转录水平在YD和AD根中以时间依赖性方式升高。免疫沉淀和Western分析表明，随着Al处理时间的增加，YD根中VHA2的磷酸化和与vf14-3-3b蛋白的相互作用得到增强，而AD根中则没有。镰刀霉素或腺苷5'-单磷酸增加或减少磷酸化VHA2和vf14-3-3b蛋白之间的相互作用，在Al胁迫条件下，两个品种的PMH（）-ATPase活性和柠檬酸盐渗出均增强或降低，分别。一起来看,这些结果表明Al增强了PMH（）-ATPase与14-3-3蛋白的表达和相互作用，从而导致PMH（）-ATPase的活性更高，并且从YD植物中渗出更多的柠檬酸盐。