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Metallothionein-like gene from Cicer microphyllum is regulated by multiple abiotic stresses

Cicer microphyllum, a wild relative of cultivated chickpea, is a high altitude cold desert-adapted species distributed in western and trans-Himalayas. A complementary DNA (cDNA) encoding metallothionein-like protein has been identified from a cold-induced subtraction cDNA library from C. microphyllum. The sequence of the cloned metallothionein gene from C. microphyllum (GQ900702) contains 240-bp-long open reading frame and encodes predicted 79-amino acid protein of 7.9 kDa. Sequence analysis identified the motifs characteristic of type II metallothionein and designated as CmMet-2. Southern hybridization confirms a single copy of the CmMet-2 gene in C. microphyllum genome. In situ hybridization indicated spatial transcript regulation of CmMet-2 in root and aerial parts and also confirmed through real-time PCR-based quantitative transcript analysis. The data revealed a significantly low level of transcript in the aerial parts than the roots. Quantitative analysis using real-time PCR assay revealed induction of transcript in all parts of plants in response to cold stress at 4°C. The transcript abundance was found to increase exponentially with time course from 6 to 24 h after exposure. Further, regulation of transcript accumulation in response to abscisic acid application, polyethylene glycol (100 μM)-induced osmotic stress, or ZnSO4 (1 μM) foliar spray indicated by Northern hybridization suggests the involvement of CmMet-2 in multiple stress response.

Authors:   Rupesh K. Singh, Sivalingam Anandhan, Shweta Singh, Vikas Y. Patade, Zakwan Ahmed, Veena Pande
Journal:   Protoplasma
Year:   2010
DOI:   10.1007/s00709-010-0249-y
Publication date:   15-Dec-2010
Facts, background information, dossiers
  • PCR assays
  • PCR
  • ?-amino acids
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