title: Metallothionein-like gene from Cicer microphyllum is regulated by multiple abiotic stresses.
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.
authors.affiliation: Molecular Biology and Genetic Engineering, Defence Institute of Bio Energy Research, Goraparao, Haldwani, Nainital, 263139, Uttarakhand, India.
authors.forename: Rupesh K
authors.initials: RK
authors.lastname: Singh
authors.forename: Sivalingam
authors.initials: S
authors.lastname: Anandhan
authors.forename: Shweta
authors.initials: S
authors.lastname: Singh
authors.forename: Vikas Y
authors.initials: VY
authors.lastname: Patade
authors.forename: Zakwan
authors.initials: Z
authors.lastname: Ahmed
authors.forename: Veena
authors.initials: V
authors.lastname: Pande
mesh_terms: D000040:Abscisic Acid / Q000494:pharmacology; D000595:Amino Acid Sequence; D015152:Blotting, Northern; D029863:Cicer / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D003080:Cold Temperature; D018744:DNA, Plant / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D017404:In Situ Hybridization, Fluorescence; D008668:Metallothionein / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009997:Osmotic Pressure; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism; D011092:Polyethylene Glycols / Q000494:pharmacology; D013312:Stress, Physiological; D013997:Time Factors; D019287:Zinc Sulfate / Q000494:pharmacology
chemical_list: D018744:DNA, Plant; D011092:Polyethylene Glycols; D000040:Abscisic Acid; D019287:Zinc Sulfate; D008668:Metallothionein