RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice.
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 ri...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3718813?pdf=render |
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| author | Huiying Zhang Xiangli Niu Jia Liu Fangming Xiao Shuqing Cao Yongsheng Liu |
| author_facet | Huiying Zhang Xiangli Niu Jia Liu Fangming Xiao Shuqing Cao Yongsheng Liu |
| author_sort | Huiying Zhang |
| collection | DOAJ |
| description | 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. |
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| language | English |
| last_indexed | 2024-04-12T09:51:17Z |
| publishDate | 2013-01-01 |
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| spelling | doaj.art-a913398446324d6e93dfd80a2539e5a82022-12-22T03:37:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0187e6904610.1371/journal.pone.0069046RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice.Huiying ZhangXiangli NiuJia LiuFangming XiaoShuqing CaoYongsheng LiuStomatal 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.http://europepmc.org/articles/PMC3718813?pdf=render |
| spellingShingle | Huiying Zhang Xiangli Niu Jia Liu Fangming Xiao Shuqing Cao Yongsheng Liu RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice. PLoS ONE |
| title | RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice. |
| title_full | RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice. |
| title_fullStr | RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice. |
| title_full_unstemmed | RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice. |
| title_short | RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice. |
| title_sort | rnai directed downregulation of vacuolar h atpase subunit a results in enhanced stomatal aperture and density in rice |
| url | http://europepmc.org/articles/PMC3718813?pdf=render |
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