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Ectopic expression of an osmotin gene leads to enhanced salt tolerance in transgenic chilli pepper (Capsicum annum L.)

  • Kondeti Subramanyam
  • K. V. Sailaja
  • Koona Subramanyam
  • D. Muralidhara Rao
  • K. LakshmideviEmail author
Original Paper

Abstract

Plants, when exposed to abiotic or biotic stress, produce several pathogenesis-related proteins to counteract the effects of stress. Osmotin is one of the important pathogenesis-related proteins induced during several stress conditions. We have developed improved salt stress tolerant transgenic chilli pepper plants (Capsicum annum L. var. Aiswarya 2103) by ectopic expression of the Nicotiana tabaccum osmotin gene using Agrobacterium tumefaciens EHA105 as a vector. Four-week-old chilli pepper leaves were used as an explant and A. tumefaciens EHA105 harboring pBINASCOSM plasmid that contains osmotin gene under the control of CaMV 35S promoter and npt II as a selectable marker was used in co-cultivation. Transgene integration and expression were analyzed using molecular, immunochemical, and biochemical assays. PCR and Southern blot analysis confirmed that osmotin gene has been successfully integrated into the genome of chilli pepper plants. The osmotin gene was stably segregated and expressed in T2 generation transgenic chilli pepper plants, and it was confirmed by Western blot analysis. Biochemical assays of these putative transgenic plants revealed enhanced levels of chlorophyll, proline, glycinebetaine, APX, SOD, DHAR, MDHAR, GR, and relative water content. Yield potential of the putative transgenic chilli pepper plants was evaluated under salinity stress conditions in a green house. The putative transgenic chilli pepper plants overexpressing the osmotin gene were morphologically similar to wild-type plants and produced 3.32 kg chilli pepper fruits per plant at 300 mM NaCl concentration.

Keywords

Capsicum annum L. Agrobacterium tumefaciens EHA105 Osmotin gene Binary vector CaMV 35S promoter Western blotting 

Abbreviations

CaMV 35S

Cauliflower mosaic virus 35S promoter

APX

Ascorbate peroxidase

SOD

Superoxide dismutase

DHAR

Dehydro ascorbate reductase

MDHAR

Monodehydroascorbate reductase

GR

Glutathione reductase

MS

Murashige and Skoog medium

BA

6-Benzyladenine

IAA

Indole-3-acetic acid

GA3

Gibberellic acid

IBA

Indole-3-butyric acid

AgNO3

Silver nitrate

Notes

Acknowledgments

We thank Sri Krishnadevaraya University, Anantapur, Andhra Pradesh, India for providing financial support to carry out the present work. The authors are grateful to Prof. M.V. Rajam, Department of Genetics, Delhi University—South campus, India for the critical correction and evaluation of this manuscript.

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Kondeti Subramanyam
    • 1
  • K. V. Sailaja
    • 1
  • Koona Subramanyam
    • 2
  • D. Muralidhara Rao
    • 3
  • K. Lakshmidevi
    • 1
    Email author
  1. 1.Department of BiochemistrySri Krishnadevaraya UniversityAnantapurIndia
  2. 2.Department of BiotechnologySreenidhi Institute of Science and Technology (Autonomous), Jawaharlal Nehru Technological UniversityHyderabadIndia
  3. 3.Department of BiotechnologySri Krishnadevaraya UniversityAnantapurIndia

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