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Molecular Breeding

, Volume 22, Issue 2, pp 169–181 | Cite as

Salt stress alleviation in transgenic Vigna mungo L. Hepper (blackgram) by overexpression of the glyoxalase I gene using a novel Cestrum yellow leaf curling virus (CmYLCV) promoter

  • Prasanna Bhomkar
  • Chandrama P. Upadhyay
  • Mukesh Saxena
  • Annamalai Muthusamy
  • N. Shiva Prakash
  • Mikhail Pooggin
  • Thomas Hohn
  • Neera B. Sarin
Article

Abstract

A reproducible and efficient transformation system utilizing the nodal regions of embryonal axis of blackgram (Vigna mungo L. Hepper) has been established via Agrobacterium tumefaciens. This is a report of genetic transformation of Vigna mungo for value addition of an agronomic trait, wherein the gene of interest, the glyoxalase I driven by a novel constitutive Cestrum yellow leaf curling viral promoter has been transferred for alleviating salt stress. The overexpression of this gene under the constitutive CaMV 35S promoter had earlier been shown to impart salt, heavy metal and drought stress tolerance in the model plant, tobacco. Molecular analyses of four independent transgenic lines performed by PCR, Southern and western blot revealed the stable integration of the transgene in the progeny. The transformation frequency was ca. 2.25% and the time required for the generation of transgenic plants was 10–11 weeks. Exposure of T1 transgenic plants as well as untransformed control plants to salt stress (100 mM NaCl) revealed that the transgenic plants survived under salt stress and set seed whereas the untransformed control plants failed to survive. The higher level of Glyoxalase I activity in transgenic lines was directly correlated with their ability to withstand salt stress. To the best of our knowledge this is the only report of engineering abiotic stress tolerance in blackgram.

Keywords

Cestrum yellow leaf curling virus (CmYLCV) promoter glyoxalase I gene (gly ISalt stress tolerance Embryonal axis Transgenic blackgram 

Abbreviations

gly I

Glyoxalase I gene

BAP

6-Benzylaminopurine

IBA

Indole butyric acid

nptII

Neomycin phosphotransferase II gene

Notes

Acknowledgements

This research project has been implemented with financial contributions from Swiss Agency for Development and Cooperation (SDC), Government of Switzerland and the Department of Biotechnology, Government of India under Indo-Swiss collaboration in Biotechnology. The gift of the gly 1 gene from Prof. S.K. Sopory of I.C.G.E.B. India and the critical evaluation of this work by Prof. Barbara Hohn (F.M.I. Basel, Switzerland) and Prof. K. Veluthambi (M.K.U. Madurai, Tamilnadu, India) is gratefully acknowledged. We are thankful to Dr. Mohd. Aslam Yusuf, J.N.U. for his valuable critical comments. The awards of Senior Research Fellowship to PB and CPU by UGC and DBT, and the experimental inputs by Ms. Nidhi Sharma, Mr. Ravi Rajwanshi and Mr. Nishakant are gratefully acknowledged.

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Prasanna Bhomkar
    • 1
  • Chandrama P. Upadhyay
    • 1
  • Mukesh Saxena
    • 1
  • Annamalai Muthusamy
    • 1
  • N. Shiva Prakash
    • 1
  • Mikhail Pooggin
    • 2
  • Thomas Hohn
    • 2
  • Neera B. Sarin
    • 1
  1. 1.School of Life SciencesJawaharlal Nehru UniversityNew DelhiIndia
  2. 2.Botanical InstituteUniversity of BaselBaselSwitzerland

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