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Plant Molecular Biology Reporter

, Volume 30, Issue 4, pp 894–903 | Cite as

Molecular Cloning and Characterization of γ-Glutamyl Cysteine Synthetase (VrγECS) from Roots of Vigna radiata (L.) Wilczek Under Progressive Drought Stress and Recovery

  • Debashree Sengupta
  • Golla Ramesh
  • Shalini Mudalkar
  • Koppolu Raja Rajesh Kumar
  • Pulugurtha Bharadwaja Kirti
  • Attipalli R. Reddy
Original Paper

Abstract

Glutathione is an essential redox buffer and an antioxidant in majority of higher plants, imparting tolerance against abiotic stress. The rate-limiting enzyme, gamma-glutamyl cysteine synthetase (γECS), plays an important role in regulation of glutathione biosynthesis under adverse environmental conditions including drought. To understand the role of γECS in an economically important food legume, Vigna radiata (L.) Wilczek, under progressive drought stress, we cloned and derived the full-length cDNA sequence and denoted it as VrγECS. Real-time PCR analysis of VrγECS in the roots of V. radiata, during progressive drought stress and recovery, indicated a stable expression pattern of the gene. However, the VrγECS enzyme activity altered differentially during varying water-deficit conditions and recovery period, reflecting the existence of some post-transcriptional or post-translational regulatory system for the enzyme. Linear regression analysis between H2O2 and lipid peroxidation as well as H2O2 and VrγECS enzyme activity during drought stress and recovery demonstrates the delicate inter-relationships and putative regulatory mechanisms operating in the root system under adverse conditions. The present study could contribute towards understanding the complex regulation of γECS in glutathione biosynthesis in an important food legume under drought stress.

Keywords

Drought Gamma-glutamyl cysteine synthetase Glutathione Mungbean Regulatory mechanisms 

Abbreviations

DAS

Days after onset of stress treatment

DAR

Days after re-watering

GSH

Glutathione

PPFD

Photosynthetic photon flux density

RACE

Rapid amplification of cDNA ends

ROS

Reactive oxygen species

γECS

Gamma-glutamyl cysteine synthetase

Notes

Acknowledgements

We thank Prof. N. Nadarajan, Tamil Nadu Agricultural University (TNAU), Coimbatore, India, for providing Vigna radiata seeds. We also thank DST-FIST facility of our department and CREBB facility of School of Life Sciences. DS and SM acknowledge the fellowship from Council of Scientific and Industrial Research (CSIR) and University Grant Commission (UGC), New Delhi, India, respectively. KRRK was supported by CSIR and Dr. D.S. Kothari Postdoctoral fellowship from UGC.

Supplementary material

11105_2011_398_Fig5_ESM.jpg (213 kb)
Supplementary Fig. S1

The complete cDNA sequence of pTZ57RT-encoded Vigna radiata γECS (JPEG 212 kb)

11105_2011_398_MOESM1_ESM.tif (149 kb)
High resolution image (TIFF 148 kb)
11105_2011_398_Fig6_ESM.jpg (321 kb)
Supplementary Fig. S2

Multiple alignment of VrγECS precursor protein with γECS of P. vulgaris, P. sativum, A. thaliana, B. napus, B. juncea and R. communis. The alignment clearly demonstrates the conservation of the cleavage site (IVAA) of the chloroplast target peptide among different plants (JPEG 321 kb)

11105_2011_398_MOESM2_ESM.tif (613 kb)
High resolution image (TIFF 613 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Debashree Sengupta
    • 1
  • Golla Ramesh
    • 2
  • Shalini Mudalkar
    • 1
  • Koppolu Raja Rajesh Kumar
    • 1
  • Pulugurtha Bharadwaja Kirti
    • 1
  • Attipalli R. Reddy
    • 1
  1. 1.Department of Plant Sciences, School of Life SciencesUniversity of HyderabadHyderabadIndia
  2. 2.Division of Haematology and Oncology, School of MedicineUniversity of PennsylvaniaPhiladelphiaUSA

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