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A reliable protocol for transformation of Catharanthus roseus through Agrobacterium tumefaciens

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Abstract

Proliferation of axillary shoot buds and multiple shoot formation in Catharanthus roseus was obtained in 96 % explants on MS medium (3 % sucrose) containing NAA + BA. 2,4-D induced callusing in both, the nodal as well as in leaf segments. Leaf-derived callus was used for transformation with Agrobacterium tumefaciens LBA4404/pBI-S1. Bacterial cell concentration, duration of co-cultivation and acetosyringone concentration influenced transformation efficiency. Under optimal co-cultivation conditions, 98 % of the explants showed GUS expression. PCR based amplification of the transformed and subsequently selected callus tissue indicated the presence of uidA, Gly I and nptII genes.

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Abbreviations

AA:

1-naphthaleneacetic acid

BA:

N6-benzyladenine

CH:

Casein hydrolysate

IAA:

Indole-3-acetic acid

Kn:

Kinetin

2,4-D:

2,4-dichlorophenoxyacetic acid

RAPD:

Random amplification of polymorphic DNA

PCR:

Polymerase chain reaction

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Authors and Affiliations

  1. Department of Biotechnology, Jamia Hamdard, New Delhi, 110062, India

    Toolika Srivastava, Sandip Das & P. S. Srivastava

  2. Plant Molecular Biology, ICGEB, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Sudhir Kumar Sopory

Authors
  1. Toolika Srivastava
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  2. Sandip Das
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  3. Sudhir Kumar Sopory
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  4. P. S. Srivastava
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Correspondence to P. S. Srivastava.

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Srivastava, T., Das, S., Sopory, S.K. et al. A reliable protocol for transformation of Catharanthus roseus through Agrobacterium tumefaciens . Physiol Mol Biol Plants 15, 93–98 (2009). https://doi.org/10.1007/s12298-009-0010-1

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