Journal Article

Functional Analysis of Water Channels in Barley Roots

Maki Katsuhara, Yoshiko Akiyama, Kazuki Koshio, Mineo Shibasaka and Kunihiro Kasamo

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 43, issue 8, pages 885-893
Published in print August 2002 | ISSN: 0032-0781
Published online August 2002 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcf102
Functional Analysis of Water Channels in Barley Roots

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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We identified three genes homologous to water channels in the plasma membrane type subfamily from roots of barley seedlings. These genes were designated HvPIP2;1, HvPIP1;3, and HvPIP1;5 after comparison to Arabidopsis aquaporins. Competitive reverse transcription (RT)-PCR was applied in order to distinguish and to quantify their transcripts. The HvPIP2;1 transcript was the most abundant among the three in roots. Salt stress (200 mM NaCl) down-regulated HvPIP2;1 (transcript and protein), but had almost no effect on the expressions of HvPIP1;3, or HvPIP1;5. Approximately equal amounts of the transcripts of the three were detected in shoots, and salt stress enhanced the expression of HvPIP2;1 but not of HvPIP1;3, or HvPIP1;5. HvPIP2;1 protein was confirmed to be localized in the plasma membrane. Functional expression of HvPIP2;1 in Xenopus oocytes confirmed that HvPIP2;1 encoded an aquaporin that transports water. This water permeability was reduced by HgCl2, which is a typical water channel inhibitor. This activity was not modified by some inhibitors against protein kinase and protein phosphatase.

Keywords: Keywords: Aquaporin — Barley — Competitive RT-PCR — Salt stress.; Abbreviations: DTT, dithiothreitol; DMSO, dimethylsulfoxide; IBMX, 3-isobutyle-1-methylxanthine; MBS, modified Barth’s solution; Pf, Osmotic water permeability; PIP, Plasma membrane Intrinsic Protein; PIPES, piperazine-N,N′-bis(2-ethanesulfonic acid); PMSF, phenylmethylsulfonylfluoride; RACE, rapid amplification of cDNA end; RT-PCR, reverse transcription-PCR; TIP, Tonoplast Intrinsic Protein.

Journal Article.  5497 words.  Illustrated.

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

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