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Genetic transformation of chickpea (Cicer arietinum L.) with insecticidal crystal protein gene using particle gun bombardment

  • Genetic Transformation and Hybridization
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Abstract

Here, we report the establishment of an efficient particle gun bombardment mediated genetic transformation in chickpea (Cicer arietinum L.) using cryIAc gene of Bacillus thuringiensis. Explants were bombarded with recombinant plasmids engineered for the expression of cryIAc transgene in plants and stable transformants regenerated in presence of benzyladenine, kinetin and kanamycin. Transformation frequency showed dependence on explant type, cultivars, plasmids, helium pressure and microcarrier type used. Integration of transgenes was demonstrated using polymerase chain reaction and Southern blot hybridization approaches in T 0 plants. The expression of CryIA(c) delta-endotoxin and GUS enzyme was ascertained by enzyme linked immunosorbent assay and histochemical assays, respectively. These transgenic plants (T 0) showed more protection and high mortality for Heliothis armigera and Spodoptera litura larvae as compared to control plants. The results of the present study indicate that highest transformation frequency (18%) could be achieved by use of gold as a microcarrier in combination with helium pressure of 900 psi. Among the other factors tested, plasmid pHS 102 was the most efficient plasmid, while epicotyl explant was the best explant source for particle gun bombardment. Among the different cultivars of chickpea tested, cultivar ICCC37 and PG-12 produced higher frequency of transformation frequency compared to others.

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Abbreviations

BA:

Benzyladenine

ELISA:

Enzyme linked immunosorbent assay

IBA:

Indole butyric acid

Kn:

Kinetin

MS:

Murashige and Skoog’s medium

PCR:

Polymerase chain reaction

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Acknowledgements

The authors are thankful to Dr. S.F. D’Souza and Dr. S.K. Apte for their encouragement, ICRISAT, AP, and MPKV, Rahuri for providing the seed material. S. Indurker is grateful to UGC for the fellowship.

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Authors and Affiliations

  1. Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai, 400 085, Maharashtra, India

    Shivani Indurker & Susan Eapen

  2. Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai, 400 085, Maharashtra, India

    Hari S. Misra

Authors
  1. Shivani Indurker
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  2. Hari S. Misra
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  3. Susan Eapen
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Corresponding author

Correspondence to Susan Eapen.

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Communicated by G. C. Phillips

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Indurker, S., Misra, H.S. & Eapen, S. Genetic transformation of chickpea (Cicer arietinum L.) with insecticidal crystal protein gene using particle gun bombardment. Plant Cell Rep 26, 755–763 (2007). https://doi.org/10.1007/s00299-006-0283-6

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  • DOI: https://doi.org/10.1007/s00299-006-0283-6

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