cDNA sequence encoding metallothionein protein from Aegiceras corniculatum and its gene expression induced by Pb²⁺ and Cd²⁺ stresses

Abstract

Constructing various green wetland examples for mangrove wetland systems is a useful way to use natural power to remediate the polluted wetlands at intertidal zones. Metallothioneins (MT) are involved in heavy metal tolerance, homeostasis, and detoxification of intracellular metal ions in plants. In order to understand the mechanism of heavy metal uptake in Aegiceras corniculatum, we isolated its metallothionein gene and studied the MT gene expression in response to heavy metals contamination. Here, we report the isolation and characterization of MT2 genes from young stem tissues of A. corniculatum growing in the cadmium (Cd) and lead (Pb) polluted wetlands of Quanzhou Bay, southeast of China. The obtained cDNA sequence of MT is 512 bp in length, and it has an open reading frame encoding 79 amino acid residues with a molecular weight of 7.92 kDa and the theoretical isoelectric point of 4.55. The amino acids include 14 cysteine residues and 14 glycine residues. It is a non-transmembrane hydrophilic protein. Sequence and homology analysis showed the MT protein sequence shared more than 60 % homology with other plant type 2 MT-like protein genes. The results suggested that the expression level of MT gene of A. corniculatum young stems induced by a certain range concentration of Cd²⁺ and Pb²⁺ stresses (0.2 mmol L⁻¹ Pb²⁺, 1 mmol L⁻¹ Pb²⁺, 0.2 mmol L⁻¹ Pb²⁺, and 40 μmmol L⁻¹ Cd²⁺; 1 mmol L⁻¹ Pb²⁺ and 40 μmol L⁻¹ Cd²⁺) compared with control might show an adaptive protection. The expression levels of MT gene at 20 h stress treatment were higher than those at 480 h stress treatment. The expression levels of MT gene with 0.2 mmol L⁻¹ Pb²⁺ stress treatment were higher than those with 0.2 mmol L⁻¹ Pb²⁺ and 40 μmol L⁻¹ Cd²⁺ stress treatment, and the MT gene expression levels with 1 mmol L⁻¹ Pb²⁺ treatment were higher than those with 1 mmol L⁻¹ Pb²⁺ and 40 μmol L⁻¹ Cd²⁺ treatment. There exists an antagonistic action between Pb²⁺ and Cd²⁺ in the MT metabolization of A. corniculatum.