Molecular Cloning and Expressional Analysis of Metallothionein Gene DaMT2a from Dioscorea alata
Metallothioneins(MTs),a class of small,cysteine(Cys)-rich and metal-binding proteins,play important roles in stress response.In this study,the full-length cDNAs encoding a metallothionein gene was obtained by RT-PCR in Dioscorea alata,which is homologous to MT2a and named as DaMT2a.Sequence analysis revealed that DaMT2a was 540 bp long,contained 3 exons and 2 introns,predicted to encode proteins of 79 amino acids,and had the conserved domains of the metallothionein.Phylogenetic analysis showed that DaMT2a was clustered into Type2 of the metallothionein family.The DaMT2a gene was explored by RT-qPCR for its expressional patterns in different tissues and in tissue-cultured seedling responding to stresses.The results indicated that DaMT2a showed a predominance expression in the stems and male flowers.The treatments of mechanical wounding,ethylene and ab-scisic acid markedly up-regulated the expression of DaMT2a,whereas the high-temperature treatment down-regu-lated its expression.This study provides a reference for the screening and functional identification of stress-related MT genes in Dioscorea alata.
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