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Plant and Cell Physiology Advance Access originally published online on July 18, 2007
Plant and Cell Physiology 2007 48(8):1081-1091; doi:10.1093/pcp/pcm091
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Rapid Paper

An Aluminum-Activated Citrate Transporter in Barley

Jun Furukawa1, Naoki Yamaji1, Hua Wang1, Namiki Mitani1, Yoshiko Murata2, Kazuhiro Sato1, Maki Katsuhara1, Kazuyoshi Takeda1 and Jian Feng Ma1,*

1Research Institute for Bioresources, Okayama University, Chuo, Kurashiki, Okayama, 710-0046 Japan
2Suntory Institute for Bioorganic Research, Wakayamadai, Shimamoto-cho, Mishima-gun, Osaka, 618-8503 Japan

*Corresponding author: E-mail, maj{at}rib.okayama-u.ac.jp; Fax, +81-86-434-1209.


  Abstract

Soluble ionic aluminum (Al) inhibits root growth and reduces crop production on acid soils. Al-resistant cultivars of barley (Hordeum vulgare L.) detoxify Al by secreting citrate from the roots, but the responsible gene has not been identified yet. Here, we identified a gene (HvAACT1) responsible for the Al-activated citrate secretion by fine mapping combined with microarray analysis, using an Al-resistant cultivar, Murasakimochi, and an Al-sensitive cultivar, Morex. This gene belongs to the multidrug and toxic compound extrusion (MATE) family and was constitutively expressed mainly in the roots of the Al-resistant barley cultivar. Heterologous expression of HvAACT1 in Xenopus oocytes showed efflux activity for 14C-labeled citrate, but not for malate. Two-electrode voltage clamp analysis also showed transport activity of citrate in the HvAACT1-expressing oocytes in the presence of Al. Overexpression of this gene in tobacco enhanced citrate secretion and Al resistance compared with the wild-type plants. Transiently expressed green fluorescent protein-tagged HvAACT1 was localized at the plasma membrane of the onion epidermal cells, and immunostaining showed that HvAACT1 was localized in the epidermal cells of the barley root tips. A good correlation was found between the expression of HvAACT1 and citrate secretion in 10 barley cultivars differing in Al resistance. Taken together, our results demonstrate that HvAACT1 is an Al-activated citrate transporter responsible for Al resistance in barley.

Keywords: Aluminum - Barley - Citrate transporter - MATE - Resistance - Root

Abbreviations: BAC, bacterial artificial chromosome; CaMV, cauliflower mosaic virus; EST, expressed sequence tag; GFP, green fluorescent protein; MATE, multidrug and toxic compound extrusion; ORF, open reading frame; QTL, quantitative trait locus; SNP, single nucleotide polymorphism

(Received June 26, 2007; Accepted July 9, 2007)
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