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

• Published in: Plant physiology and biochemistry Vol. 70; pp. 504 - 511
• Main Authors: Chen, Qi, Guo, Chuan-Long, Wang, Ping, Chen, Xuan-Qin, Wu, Kong-Huan, Li, Kui-Zhi, Yu, Yong-Xiong, Chen, Li-Mei
• Format: Journal Article
• Language: English
• Published: Paris Elsevier Masson SAS 01.09.2013; Elsevier
• Subjects: 14-3-3 Protein; 14-3-3 Proteins - metabolism; Adaptation, Physiological - genetics; adenosine; Adenosine Monophosphate - pharmacology; Al stress; aluminum; Aluminum - adverse effects; Aluminum - metabolism; Biological and medical sciences; Broad bean; Cell Membrane - metabolism; Citrate exudation; citrates; Citric Acid - metabolism; correlation; cultivars; excretion; exudation; faba beans; Fundamental and applied biological sciences. Psychology; fusicoccin; gene expression regulation; Gene Expression Regulation, Plant; genes; Genes, Plant; Glycosides - pharmacology; H-transporting ATP synthase; H-transporting ATPase; Phosphorylation; Plant physiology and development; Plant Proteins - genetics; Plant Proteins - metabolism; plasma membrane; Plasma membrane H+-ATPase; precipitin tests; Proton-Translocating ATPases - genetics; Proton-Translocating ATPases - metabolism; protons; reverse transcriptase polymerase chain reaction; roots; secretion; Stress, Physiological - genetics; Transcription, Genetic - drug effects; Up-Regulation; Vicia faba; Vicia faba - drug effects; Vicia faba - genetics; Vicia faba - metabolism; vf14-3-3b; AD; 14-3-3 Protein; Plasma membrane H+-ATPase; Al; YD; 5’-AMP; VHA2p; Al stress; OA; Broad bean; vha2; DDT; PMSF; Citrate exudation; FC; PM; BTP; Enzyme; Interaction; ATPase; Citrate; H; Aluminium; transporting ATP synthase; Vicia faba; Protein; Grain legume; Phytotoxicity; Leguminosae; Plant physiology; Dicotyledones; Angiospermae; Plasma membrane; Plasma membrane H; Hydrolases; Spermatophyta; Exudation; Vicia faba plasma membrane H(+)-ATPase; aluminum; phenylmethylsulfonyl fluoride; fusicoccin; phosphorylated VHA2 at Thr-951 in C terminus; 1,3-bis-[tris(hy- droxymethyl)-methylamino]-propane; dithiothreitol; plasma membrane; Vicia faba 14-3-3-like protein b; Vicia faba. L cv. Yunnan Candou 8363; adenosine 5′-monophosphate; Plasma membrane H(+)-ATPase; Vicia faba. L cv. Anhui Candou; organic acid
• ISSN: 0981-9428; 1873-2690
• DOI: 10.1016/j.plaphy.2013.06.015

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. [Display omitted] •We analyzed the regulatory mechanism of Al-induced citrate exudation in broad bean.•More citrate exudation is observed in Al-resistant YD than sensitive AD.•PM H+-ATPase activity was positively related to citrate exudation in YD and AD.•Higher transcription amounts of VHA2 and vf14-3-3b were detected in YD than AD.•The interaction between VHA2p and the vf14-3-3b was enhanced in YD but not in AD.