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Plant and Cell Physiology Advance Access originally published online on November 18, 2008
Plant and Cell Physiology 2009 50(1):106-117; doi:10.1093/pcp/pcn175
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Novel Cysteine-Rich Peptides from Digitaria ciliaris and Oryza sativa Enhance Tolerance to Cadmium by Limiting its Cellular Accumulation

Masato Kuramata1,4, Shuichi Masuya1, Yoshihiro Takahashi1, Etsuko Kitagawa2, Chihiro Inoue3, Satoru Ishikawa4, Shohab Youssefian5 and Tomonobu Kusano1,*

1Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Aoba, Sendai, Miyagi, 980-8577 Japan
2Akita Agricultural Experiment Station, Akita Prefectural Agriculture, Forestry and Fisheries Research Center, 34-1 Aikawa, Yuwa, Akita, 010-1231 Japan
3Graduate School of Environmental Studies, Tohoku University, 6-6-20 Aoba, Aramaki, Aoba, Sendai, Miyagi, 980-8579 Japan
4Department of Environmental Chemistry, National Institute for Agro-Environmental Sciences, 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-0856 Japan
5Faculty of Biological Resources, Akita Prefectural University, 241-7 Kaidobata Nishi, Akita, 010-1095 Japan

*Corresponding author: E-mail, kusano{at}ige.tohoku.ac.jp;Fax, 81-22-217-5709.


  Abstract

By means of functional screening using the cadmium (Cd)-sensitive ycf1 yeast mutant, we have isolated a novel cDNA clone, DcCDT1, from Digitaria ciliaris growing in a former mining area in northern Japan, and have shown that it confers Cd tolerance to the yeast cells, which accumulated almost 2-fold lower Cd levels than control cells. The 521 bp DcCDT1 cDNA contains an open reading frame of 168 bp and encodes a deduced peptide, DcCDT1, that is 55 amino acid residues in length, of which 15 (27.3%) are cysteine residues. Five DcCDT1 homologs (here termed OsCDT1–OsCDT5) have been identified in rice, and all of them were up-regulated to varying degrees in the above-ground tissues by CdCl2 treatment. Localization of green fluorescent protein fusions suggests that DcCDT1 and OsCDT1 are targeted to both cytoplasmic membranes and cell walls of plant cells. Transgenic Arabidopsis thaliana plants overexpressing DcCDT1 or OsCDT1 displayed a Cd-tolerant phenotype and, consistent with our yeast data, accumulated lower amounts of Cd when grown on CdCl2. Collectively, our data suggest that DcCDT1 and OsCDT1 function to prevent entry of Cd into yeast and plant cells and thereby enhance their Cd tolerance.

Keywords: Cadmium - Cysteine-rich peptide - Digitaria ciliaris - Oryza sativa - Tolerance - Transgenic plant

Abbreviations: Cd, cadmium; GFP, green fluorescent protein; ICP-MS, inductively coupled plasma-mass spectrometry; MT, metallothionein; PC, phytochelatin; RT–PCR, reverse transcription–PCR; YCF1, yeast cadmium factor 1.

The nucleotide sequences described here have been deposited in the GenBank/DDBJ database under the following accession numbers: DcCDT1, AB333802 [GenBank] ; DcMT1, AB333803 [GenBank] ; EcCDT1, AB426477 [GenBank] ; MsCDT1, AB426478.

(Received August 26, 2008; Accepted November 13, 2008)
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