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Characterization of a novel glycinebetaine/proline transporter gene expressed in the mestome sheath and lateral root cap cells in barley

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Abstract

The accumulation of glycinebetaine (GB) is one of the adaptive strategies to adverse salt stress conditions. Although it has been demonstrated that barley plants accumulate GB in response to salt stress and various studies focused on GB synthesis were performed, its transport mechanism is still unclear. In this study, we identified a novel gene, HvProT2, encoding Hordeum vulgare GB/proline transporter from barley plants. Heterologous expression in yeast (Saccharomyces cerevisiae) mutant demonstrated that the affinity of HvProT2 was highest for GB, intermediate for proline and lowest for γ-aminobutyric acid. Transient expression of fusions of HvProT2 and green fluorescent protein in onion epidermal cells revealed that HvProT2 is localized at the plasma membrane. Relative quantification of mRNA level of HvProT2 using semi-quantitative reverse transcription-polymerase chain reaction analysis showed that HvProT2 is constitutively expressed in both leaves and roots, and the expression level was higher in old leaves than young leaves and roots. Moreover, we found that HvProT2 was expressed in the mestome sheath and lateral root cap cells. We discussed the possible involvement of HvProT2 for salt stress tolerance.

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Abbreviations

BADH:

Betaine aldehyde dehydrogenase

BBD:

Barley betaine aldehyde dehydrogenase

CaMV:

Cauliflower mosaic virus

cDNA:

Complementary deoxyribonucleic acid

DIG:

Digoxigenin

DNA:

Deoxyribonucleic acid

DNP:

Dinitrophenol

EF:

Elongation factor

EST:

Expressed sequence tag

GABA:

γ-Aminobutyric acid

GB:

Glycinebetaine

GFP:

Green fluorescent protein

mRNA:

Messenger ribonucleic acid

ORF:

Open reading frame

P5CS:

Δ1-Pyrroline-5-carboxylate synthetase

PCR:

Polymerase chain reaction

RNA:

Ribonucleic acid

RT-PCR:

Reverse transcription-polymerase chain reaction

SD:

Synthetic dextrose

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Acknowledgments

We are grateful to Dr. Bruno André (Université Libre de Bruxelles, Belgium) for providing the yeast strain 22574d. This work was supported by Research Fellow of the Japan Society for the Promotion of Science (to T. F.), Grant-in-Aid for scientific research (Nos. 20380177 and 18880013) from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to T. T. and S. M., respectively) and The Salt Science Research Foundation (to S. M.).

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Authors and Affiliations

  1. Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya, 464-8601, Japan

    Takashi Fujiwara, Shiro Mitsuya, Hiroshi Miyake, Tasuku Hattori & Tetsuko Takabe

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  1. Takashi Fujiwara
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  2. Shiro Mitsuya
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  3. Hiroshi Miyake
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  4. Tasuku Hattori
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  5. Tetsuko Takabe
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Correspondence to Tetsuko Takabe.

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Fujiwara, T., Mitsuya, S., Miyake, H. et al. Characterization of a novel glycinebetaine/proline transporter gene expressed in the mestome sheath and lateral root cap cells in barley. Planta 232, 133–143 (2010). https://doi.org/10.1007/s00425-010-1155-4

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  • DOI: https://doi.org/10.1007/s00425-010-1155-4

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