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Plant and Cell Physiology Advance Access published online on April 17, 2008
Plant and Cell Physiology, doi:10.1093/pcp/pcn062
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Overexpression of a new rice vacuolar antiporter regulating protein OsARP improves salt tolerance in tobacco

Md. Imtiaz Uddin1, Yanhua Qi2, Satoshi Yamada3, Izumi Shibuya4, Xi-Ping Deng5, Sang-Soo Kwak6, Hironori Kaminaka1 and Kiyoshi Tanaka1

1Lab of Plant Biotechnology, Faculty of Agriculture, Tottori University, Koyama-cho, Minami 4-101, Tottori, 680-8553, Japan
2The State Key Laboratory of Plant Physiology and Biochemistry, College of Life Science, Zhejiang University, Hangzhou 310058, China
3Lab of Plant Nutrition, Faculty of Agriculture, Tottori University, Koyama-cho, Minami 4-101, Tottori 680-8553, Japan
4Department of Veterinary Physiology, Faculty of Veterinary Medicine, Tottori University, Koyama-cho, Minami 4-101, Tottori 680-8553, Japan
5The State Key Laboratory of Soil Erosion and Dryland Farming on the Loess Plateau, Institute of Soil and Water Conservation, Chinese Academy of Sciences, Yangling, Shaanxi 712100, China
6Lab of Environmental Biotechnology, Korea Research Institute of Bioscience and Biotechnology, Yuseong-gu, Daejeon 305-806, Korea

Corresponding author: Professor. Kiyoshi Tanaka, Ph. D. Address: Lab of Plant Biotechnology, Faculty of Agriculture, Tottori University, Koyama-cho, Minami 4-101, Tottori, 680-8553, JapanTel/ Fax: +81-857-31-5638, E-mail: jotanaka{at}muses.tottori-u.ac.jp


  Abstract

We examined function of the rice (Oryza sativa L.) antiporter regulating protein OsARP by overexpressing in tobacco (Nicotiana tabacum L.). In public databases, this protein was annotated as putative Os02g0465900 protein of rice. The OsARP gene was introduced into tobacco under the control of cauliflower mosaic virus 35S promoter. The transformants were selected for their ability to grow on medium containing kanamycin. Incorporation of the transgene in the genome of tobacco was confirmed by polymerase chain reaction, and its expression was confirmed by Western-blot analysis. Transgenic plants had better growth and vigor than that of non-transgenic under salt stress in vitro. Overexpression of OsARP in transgenic tobacco plants resulted in salt tolerance had higher photosynthesis and effective PSII photon yield when compared with the wild type. The OsARP protein was localized in tonoplast of rice plants. Transgenic plants accumulated more Na+ in their leaf tissue than those of wild type plants. It is conceivable that the toxic effect of Na+ in cytosol might be reduced by sequestration into vacuoles. The rate of water loss was higher in wild type than that of transgenic under salt stress. Increased vacuolar solute accumulation and water retention could confer salt tolerance in transgenic plants. Tonoplast vesicles isolated from OsARP transgenic plants showed Na+/H+ exchange rates three-fold higher than those of wild type plants. These results suggest that OsARP on the tonoplasts plays important role in compartmentation of Na+ into vacuoles. We suggest OsARP is a new type protein participating in Na+ uptake in vacuoles.

Keywords: Tobacco (Nicotiana tabacum L.) - Rice (Oryza sativa L.) - Salt tolerance - Vacuolar antiporter regulator - Na+/H+ exchange

(Received April 8, 2008; Accepted April 13, 2008)
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