Journal Article

Functional Analysis of Salt-Inducible Proline Transporter of Barley Roots

Akihiro Ueda, Weiming Shi, Kazutsuka Sanmiya, Mariko Shono and Tetsuko Takabe

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 42, issue 11, pages 1282-1289
Published in print November 2001 | ISSN: 0032-0781
Published online November 2001 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pce166
Functional Analysis of Salt-Inducible Proline Transporter of Barley Roots

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We cloned a cDNA encoding Hordeum vulgare Proline Transporter (HvProT) from salt-stressed barley roots by differential display. HvProT was 2,161 bp long and had an open reading frame encoding 450 amino acids. The deduced amino acid sequence of HvProT was similar to those of proline transporter proteins of rice (65.7%), Arabidopsis (57.7%) and tomato (42.0%). Northern blot analysis showed that the transcript level of HvProT was induced in roots at 30 min after 200 mM NaCl treatment and its peak was observed at 3 h. However, the transcript level was very low in leaves and did not increase by salt stress. The expression level of Δ1-pyrroline-5-carboxylate synthetase (P5CS), encoding a key enzyme of proline synthesis, was induced later than HvProT by salt stress. A transport assay using a yeast with mutation in proline uptake revealed that HvProT was a transporter with high affinity for l-proline (Km = 25 µM). HvProT was found to be a unique transporter with high affinity for l-proline. Since its transport activity was dependent on the pH gradient, HvProT was suggested to be a H+/amino acid symporter. In situ hybridization analysis showed that the HvProT mRNA was strongly expressed in root cap cells under salt stress. HvProT might play an important role in the transport of proline to root tip region urgently upon salt stress.

Keywords: Key words: Hordeum vulgare — Proline transporter — Salt stress — Differential display — In situ hybridization.; Abbreviations: CCCP, carbonyl cyanide m-chlorophenyl-hydrazone; DIG, digoxygenin; GABA, γ-amino butyric acid; P5CS, Δ1-pyrroline-5-carboxylate synthetase; ProT, proline transporter.

Journal Article.  3867 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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