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First published online September 3, 2004; 10.1104/pp.104.046003

Plant Physiology 136:2532-2547 (2004)
© 2004 American Society of Plant Biologists

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Expression Patterns of a Novel AtCHX Gene Family Highlight Potential Roles in Osmotic Adjustment and K+ Homeostasis in Pollen Development1,[w]

Heven Sze*, Senthilkumar Padmanaban, Françoise Cellier, David Honys, Ning-Hui Cheng, Kevin W. Bock, Genevieve Conéjéro, Xiyan Li, David Twell, John M. Ward and Kendal D. Hirschi

Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland 20742–5815 (H.S., S.P., K.W.B., X.L.); Biochimie et Physiologie Moléculaire des Plantes, Unité Mixte de Recherche 5004 Institute National de la Recherche Agronomique/Centre National de la Recherche Scientifique/AgroM/Université Montpellier II, 34060 Montpellier cedex, France (F.C., G.C.); Laboratory of Pollen Biology, Institute of Experimental Botany Academy of Sciences of the Czech Republic, 16502 Prague 6, Czech Republic (D.H.); Departments of Pediatrics and Human and Molecular Genetics, Children's Nutrition Research Center, Baylor College of Medicine, Houston, Texas 77030 (N.-H.C., K.D.H.); Department of Biology, University of Leicester, Leicester LE1 7RH, United Kingdom (D.T.); and Department of Plant Biology, University of Minnesota, St. Paul, Minnesota 55108 (J.M.W.)

A combined bioinformatic and experimental approach is being used to uncover the functions of a novel family of cation/H+ exchanger (CHX) genes in plants using Arabidopsis as a model. The predicted protein (85–95 kD) of 28 AtCHX genes after revision consists of an amino-terminal domain with 10 to 12 transmembrane spans (approximately 440 residues) and a hydrophilic domain of approximately 360 residues at the carboxyl end, which is proposed to have regulatory roles. The hydrophobic, but not the hydrophilic, domain of plant CHX is remarkably similar to monovalent cation/proton antiporter-2 (CPA2) proteins, especially yeast (Saccharomyces cerevisiae) KHA1 and Synechocystis NhaS4. Reports of characterized fungal and prokaryotic CPA2 indicate that they have various transport modes, including K+/H+ (KHA1), Na+/H+-K+ (GerN) antiport, and ligand-gated ion channel (KefC). The expression pattern of AtCHX genes was determined by reverse transcription PCR, promoter-driven {beta}-glucuronidase expression in transgenic plants, and Affymetrix ATH1 genome arrays. Results show that 18 genes are specifically or preferentially expressed in the male gametophyte, and six genes are highly expressed in sporophytic tissues. Microarray data revealed that several AtCHX genes were developmentally regulated during microgametogenesis. An exciting idea is that CHX proteins allow osmotic adjustment and K+ homeostasis as mature pollen desiccates and then rehydrates at germination. The multiplicity of CHX-like genes is conserved in higher plants but is not found in animals. Only 17 genes, OsCHX01 to OsCHX17, were identified in rice (Oryza sativa) subsp. japonica, suggesting diversification of CHX in Arabidopsis. These results reveal a novel CHX gene family in flowering plants with potential functions in pollen development, germination, and tube growth.


1 This work was supported in part by the National Science Foundation Arabidopsis 2010 Project (grant nos. IBN0209788 and IBN0200093 to H.S., 0209792 to J.M.W., and 020977 to K.H.) and by a Royal Society/NATO Fellowship and the Ministry of Education of the Czech Republic (grant no. 1K03018 to D.H.).

[w] The online version of this article contains Web-only data.

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.046003.

* Corresponding author; e-mail hsze{at}umd.edu; fax 301–314–9081.

Received May 14, 2004; returned for revision June 14, 2004; accepted July 12, 2004.




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