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

Protoplasma. 2011 Oct;248(4):839-47. doi: 10.1007/s00709-010-0249-y. Epub 2010 Dec 16.

Abstract

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 ZnSO(4) (1 μM) foliar spray indicated by Northern hybridization suggests the involvement of CmMet-2 in multiple stress response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abscisic Acid / pharmacology
  • Amino Acid Sequence
  • Blotting, Northern
  • Cicer / drug effects
  • Cicer / genetics*
  • Cicer / metabolism
  • Cloning, Molecular
  • Cold Temperature
  • DNA, Plant / genetics*
  • Gene Expression Regulation, Plant
  • Gene Library
  • In Situ Hybridization, Fluorescence
  • Metallothionein / genetics*
  • Metallothionein / metabolism
  • Molecular Sequence Data
  • Open Reading Frames
  • Osmotic Pressure
  • Plant Leaves / drug effects
  • Plant Leaves / genetics
  • Plant Leaves / metabolism
  • Plant Roots / genetics
  • Plant Roots / metabolism
  • Polyethylene Glycols / pharmacology
  • Stress, Physiological*
  • Time Factors
  • Zinc Sulfate / pharmacology

Substances

  • DNA, Plant
  • Polyethylene Glycols
  • Abscisic Acid
  • Zinc Sulfate
  • Metallothionein