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Molecular characterization of a rice metal tolerance protein, OsMTP1

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Abstract

Rice (Oryza sativa L. ‘Nipponbare’) cDNA subtractive suppression hybridization (SSH) libraries constructed using cadmium (Cd)-treated seedling roots were screened to isolate Cd-responsive genes. A cDNA clone, encoding the rice homolog of Metal Tolerance Protein (OsMTP1), was induced by Cd treatment. Plant MTPs belong to cation diffusion facilitator (CDF) protein family, which are widespread in bacteria, fungi, plants, and animals. OsMTP1 heterologous expression in yeast mutants showed that OsMTP1 was able to complement the mutant strains’ hypersensitivity to Ni, Cd, and Zn, but not other metals including Co and Mn. OsMTP1 expression increased tolerance to Zn, Cd, and Ni in wild-type yeast BY4741 during the exponential growth phase. OsMTP1 fused to green fluorescent protein was localized in onion epidermal cell plasma membranes, consistent with an OsMTP1 function in heavy metal transporting. OsMTP1 dsRNAi mediated by transgenic assay in rice seedlings resulted in heavy metal sensitivity and changed the heavy metal accumulation in different organs of mature rice under low-concentration heavy metal stress. Taken together, our results show that OsMTP1 is a bivalent cation transporter localized in the cell membrane, which is necessary for efficient translocation of Zn, Cd and other heavy metals, and maintain ion homeostasis in plant.

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Abbreviations

MTP:

Metal tolerance protein

CDF:

Cation diffusion facilitator

NRAMP:

Natural resistance-associated macrophage protein

ZIP:

Zn-regulated transporter-/Fe-regulated transporter-like protein

TMD:

Transmembrane domain

ZAT:

Zinc transporter of Arabidopsis thaliana

AAS:

Atomic absorption spectrometer

dsRNAi:

Double-stranded RNA interference

SD medium:

Synthetic defined medium

RT-PCR:

Reverse transcription PCR

DIG:

Digoxigenin

ORF:

Open reading frame

EGFP:

Enhanced green fluorescent protein

SSH:

Subtractive suppression hybridization

ISH:

In situ hybridization

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Acknowledgments

We sincerely thank Wei Zhang for her help in the rice transformation assay. We are also very grateful to Prof. David Ow for his comments and constructive suggestions to improve this manuscript. This research was supported by the China National Natural Sciences Foundation (Grant no. 30900892), the International Foundation for Science (Grant no. C/4787) to Mei Zhang and the CAS ‘100 Talents’ Program to Keqiang Wu.

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Authors and Affiliations

  1. Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, 510650, People’s Republic of China

    Lianyu Yuan, Songguang Yang, Baoxiu Liu, Mei Zhang & Keqiang Wu

  2. Graduate School of the Chinese Academy of Sciences, Beijing, 100039, People’s Republic of China

    Lianyu Yuan & Baoxiu Liu

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  1. Lianyu Yuan
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  2. Songguang Yang
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  3. Baoxiu Liu
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  4. Mei Zhang
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  5. Keqiang Wu
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Correspondence to Mei Zhang.

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Communicated by Y. Lu.

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Yuan, L., Yang, S., Liu, B. et al. Molecular characterization of a rice metal tolerance protein, OsMTP1 . Plant Cell Rep 31, 67–79 (2012). https://doi.org/10.1007/s00299-011-1140-9

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