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Characterization of Al-responsive citrate excretion and citrate-transporting MATEs in Eucalyptus camaldulensis

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Abstract

Many plant species excrete organic acids into the rhizosphere in response to aluminum stress to protect sensitive cells from aluminum rhizotoxicity. When the roots of Eucalyptus camaldulensis, a major source of pulp production, were incubated in aluminum-toxic medium, citrate released into the solution increased as a function of time. Citrate excretion was inducible by aluminum, but not by copper or sodium chloride stresses. This indicated that citrate is the major responsive organic acid released from the roots of this plant species to protect the root tips from aluminum damage. Four genes highly homologs to known citrate-transporting multidrugs and toxic compounds exclusion proteins, named EcMATE1–4, were isolated using polymerase chain reaction-based cloning techniques. Their predicted proteins included 12 membrane spanning domains, a common structural feature of citrate-transporting MATE proteins, and consisted of 502–579 amino acids with >60 % homology to orthologous genes in other plant species. One of the homologs, designated EcMATE1, was expressed in the roots more abundantly than in the shoots and in response to both Al and low pH stresses. Ectopic expression of EcMATE1 and 3 in tobacco hairy roots enhanced Al-responsive citrate excretion. Pharmacological characterization indicated that Al-responsive citrate excretion involved a protein phosphorylation/dephosphorylation process. These results indicate that citrate excretion through citrate-transporting multidrugs and toxic compounds exclusion proteins is one of the important aluminum-tolerance mechanisms in Eucalyptus camaldulensis.

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Abbreviations

Al:

Aluminum

ALMT1:

Aluminum-activated malate transporter 1

ART1:

Al-resistance transcription factor 1

AtFRD3:

Arabidopsis thaliana ferric redictase defective3

HvAACT1:

Hordeum vulgare aluminum-activated citrate transporter 1

GUS:

β-Glucuronidase

MATE:

Multidrugs and toxic compounds exclusion

MS:

Murashige and Skoog

OAs:

Organic acids

STOP1:

Sensitive to proton rhizotoxicity1

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Acknowledgments

We thank Dr Tsuyoshi Nakagawa (Shimane University) for providing the gateway vector, pGWB2. This work was supported by the METI (Ministry of Economy, Trade and Industry) Grant Program, Japan.

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Authors and Affiliations

  1. Forest Technology Laboratories, Research and Development Division, Oji Paper Co., Ltd., 1-10-6 Shinonome, Koto-ku, Tokyo, 135-8558, Japan

    Yoshiharu Sawaki, Tomonori Kihara-Doi, Nobuyuki Nishikubo, Tetsu Kawazu & Shigeru Sato

  2. Laboratory of Plant Cell Technology, Faculty of Applied Biological Sciences, Gifu University, 1-1 Yanagido, Gifu, 501-1193, Japan

    Yuriko Kobayashi, Yasufumi Kobayashi & Hiroyuki Koyama

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  1. Yoshiharu Sawaki
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  2. Tomonori Kihara-Doi
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  3. Yuriko Kobayashi
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  4. Nobuyuki Nishikubo
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  5. Tetsu Kawazu
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  6. Yasufumi Kobayashi
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  8. Shigeru Sato
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Correspondence to Shigeru Sato.

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Sawaki, Y., Kihara-Doi, T., Kobayashi, Y. et al. Characterization of Al-responsive citrate excretion and citrate-transporting MATEs in Eucalyptus camaldulensis . Planta 237, 979–989 (2013). https://doi.org/10.1007/s00425-012-1810-z

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