Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (14): 2904-2912.doi: 10.3864/j.issn.0578-1752.2012.14.013

• HORTICULTURE • Previous Articles     Next Articles

Cloning and Expression Analysis of Metallothionein Gene (MhMT2) in Malus hupehensis

 WANG  Shun-Cai, LIANG  Dong, MA  Feng-Wang   

  1. 1.天水师范学院生命科学与化学学院，甘肃天水748100
    2.西北农林科技大学园艺学院，陕西杨凌712100
  • Received:2012-03-21 Online:2012-07-15 Published:2012-05-08

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Abstract: 【Objective】The objective of this study is to clone the full-length cDNA of metallothionein (MT) gene from rootstock (Malus hupehensis), to understand the mechanism of MT in response to environmental stress and provide fundamental evidence for molecular breeding of apple trees under stress conditions.【Method】The full-length cDNA sequence of MhMT2 gene was amplified from the leaves of M. hupehensis using in silico cloning and RT-PCR. Bioinformatics methods were used to analyze cDNA sequence obtained and putative amino acid sequence. The expression profiles of MhMT2 transcript in control seedlings and those exposed to various abiotic stresses and hormone treatments were detected by quantitative real-time RT-PCR.【Result】Sequence analysis indicated that MhMT2 cDNA was 534 bp in length and contained an intact open reading frame of 243 bp, encoding a polypeptide of 80 amino acids with a theoretical molecular mass of 7.87 kD. Homology comparison analysis showed that the deduced MhMT2 protein was highly homologous to other MT2 proteins from plant species, sharing a 96.4% homology with M. xiaojinensis. Bioinformatic analysis revealed that the putative MhMT2 protein contained 14 cysteine residues occurring as C-C, C-X-C, and C-X-X-C motifs in the N-terminal, and of C-X-C in the C-terminal region (X are other amino acids other than cysteine). Phylogenetic analysis suggested that this gene belonged to type 2 plant MT genes family. Quantitative real-time RT-PCR analysis indicated that MhMT2 showed the highest transcript abundance in leaves, moderate level in roots and the lowest level in shoots. MhMT2 transcripts in leaves were markedly increased by abscisic acid (ABA), hydrogen peroxide (H2O2), mechanical wounding, and NaCl, especially H2O2 treatment.【Conclusion】The induced expression profiling of MhMT2 gene under ABA, H2O2, mechanical wounding, and salt stress indicates that the MhMT2 gene may play an important role in response to these abiotic stresses.

Key words: M. hupehensis, metallothionein gene, cloning, expression

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