title: RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice.
abstract: Stomatal movement plays a key role in plant development and response to drought and salt stress by regulating gas exchange and water loss. A number of genes have been demonstrated to be involved in the regulation of this process. Using inverse genetics approach, we characterized the function of a rice (Oryza sativa L.) vacuolar H(+)-ATPase subunit A (OsVHA-A) gene in stomatal conductance regulation and physiological response to salt and osmotic stress. OsVHA-A was constitutively expressed in different rice tissues, and the fusion protein of GFP-OsVHA-A was exclusively targeted to tonoplast when transiently expressed in the onion epidermal cells. Heterologous expression of OsVHA-A was able to rescue the yeast mutant vma1Δ (lacking subunit A activity) phenotype, suggesting that it partially restores the activity of V-ATPase. Meanwhile, RNAi-directed knockdown of OsVHA-A led to a reduction of vacuolar H(+)-ATPase activity and an enhancement of plasma membrane H(+)-ATPase activity, thereby increasing the concentrations of extracellular H(+) and intracellular K(+) and Na(+) under stress conditions. Knockdown of OsVHA-A also resulted in the upregulation of PAM3 (plasma membrane H(+)-ATPase 3) and downregulation of CAM1 (calmodulin 1), CAM3 (calmodulin 3) and YDA1 (YODA, a MAPKK gene). Altered level of the ion concentration and the gene expression by knockdown of OsVHA-A probably resulted in expanded aperture of stomatal pores and increased stomatal density. In addition, OsVHA-A RNAi plants displayed significant growth inhibition under salt and osmotic stress conditions. Taken together, our results suggest that OsVHA-A takes part in regulating stomatal density and opening via interfering with pH value and ionic equilibrium in guard cells and thereby affects the growth of rice plants.
authors.affiliation: School of Life Science, Chongqing University, Chongqing, China.
authors.forename: Huiying
authors.initials: H
authors.lastname: Zhang
authors.forename: Xiangli
authors.initials: X
authors.lastname: Niu
authors.forename: Jia
authors.initials: J
authors.lastname: Liu
authors.forename: Fangming
authors.initials: F
authors.lastname: Xiao
authors.forename: Shuqing
authors.initials: S
authors.lastname: Cao
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Liu
mesh_terms: D015536:Down-Regulation / Q000235:genetics*; D055864:Droughts; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D055785:Gene Knockdown Techniques; D006863:Hydrogen-Ion Concentration; D042541:Intracellular Space / Q000378:metabolism; D012275:Oryza / Q000166:cytology / Q000235:genetics* / Q000254:growth & development* / Q000502:physiology; D009997:Osmotic Pressure; D054046:Plant Stomata / Q000254:growth & development*; D021122:Protein Subunits / Q000172:deficiency / Q000235:genetics; D021381:Protein Transport; D034622:RNA Interference; D012492:Salts / Q000494:pharmacology; D013312:Stress, Physiological / Q000187:drug effects / Q000235:genetics; D025262:Vacuolar Proton-Translocating ATPases / Q000172:deficiency* / Q000235:genetics*
chemical_list: D021122:Protein Subunits; D012492:Salts; D025262:Vacuolar Proton-Translocating ATPases
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