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Cloning and characterization of GPAT gene from Lepidium latifolium L.: a step towards translational research in agri-genomics for food and fuel

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Abstract

Glycerol-3-phosphate acyltransferase (GPAT) catalyzes first and the rate limiting step in glycerolipid synthesis pathway, which in turn contribute to stabilization of plasma membrane structure and oil lipid synthesis in plant cells. Here, we report cloning and characterization of GPAT gene from Lepidium latifolium (LlaGPAT). The cDNA sequence (1,615 bp) of LlaGPAT gene consisted of 1,113 bp ORF encoding a protein of 370 aa residues, with deduced mass of 41.2 kDa and four acyltransferase (AT) motifs having role in catalysis and in glycerol-3-phosphate binding. Southern blot analysis suggested presence of a single copy of the gene in the genome. Tissue specific expression of the gene was seen more abundantly in aerial parts, compared to the roots. Quantitative real-time PCR indicated down-regulation of the gene by cold (4 °C), drought (PEG6000), salt (300 mM NaCl) and ABA (100 μM) treatments. Considering the vitality of the function of encoded enzyme, LlaGPAT can be considered a potential candidate gene for genetic engineering of oil yields and abiotic stress management in food as well as fuel crops.

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Abbreviations

GPAT:

Glycerol-3-phosphate acyltransferase

PEG:

Poly ethylene glycol

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Reverse transcription-PCR

TAG:

Triacylglycerol

qPCR:

Quantitative PCR

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Acknowledgments

Defence Research and Development Organisation (DRDO), HQ, New Delhi is duly acknowledged for funding the project and providing Senior Research Fellowship (SRF) to PP and SS.

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Authors and Affiliations

  1. Molecular Biology and Genetic Engineering Laboratory, Defence Institute of Bio-Energy Research (DIBER), DRDO, Goraparao, P.O.-Arjunpur, Haldwani, Nainital, 263139, Uttarakhand, India

    Sanjay Mohan Gupta, Pankaj Pandey, Atul Grover, Vikas Yadav Patade, Sadhana Singh & Zakwan Ahmed

Authors
  1. Sanjay Mohan Gupta
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  2. Pankaj Pandey
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  3. Atul Grover
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  4. Vikas Yadav Patade
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  5. Sadhana Singh
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  6. Zakwan Ahmed
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Corresponding author

Correspondence to Sanjay Mohan Gupta.

Additional information

GenBank Accession Numbers: cDNA sequence of LlaGPAT: GenBank Acc. No. JN398166; Genomic sequence of LlaGPAT: GenBank Acc. No. JN882006.

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Gupta, S.M., Pandey, P., Grover, A. et al. Cloning and characterization of GPAT gene from Lepidium latifolium L.: a step towards translational research in agri-genomics for food and fuel. Mol Biol Rep 40, 4235–4240 (2013). https://doi.org/10.1007/s11033-013-2505-7

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  • DOI: https://doi.org/10.1007/s11033-013-2505-7

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