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Transformation through agroinfection on decapitated shoot apex of field-growing Phyllanthus amarus

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Abstract

A simple and easy transformation strategy was accomplished on field growing plants of Phyllanthus amarus, an anti-hepatitis B drug plant. Infection of Agrobacterium rhizogenes strains A4M70GUS and ATCC 15834 on decapitated shoots of field growing P. amarus induced hairy roots and crown gall, respectively. Infection with A4M70GUS yielded a mean of 23.2 roots from 40% plants in 40-day period. The crown gall induced on 30% plants after infection with ATCC 15834 grew to 5–10 mm in diameter. The roots and crown galls established in vitro on Murashige and Skoog (MS) basal medium grew well. The hairy roots yielded fivefold (6.91 g) biomass in half-strength MS liquid medium to that of the adventitious roots derived from internode explants in MS medium with 8.0 μM α-naphthaleneacetic acid (1.39 g). Histochemical assay and PCR analysis using the primers of uidA coding region confirmed the hairy roots induced by A4M70GUS. The crown galls induced by ATCC 15834 were confirmed by PCR analysis using rolB gene primers. The protocol enables an easy and early accomplishment of hairy roots.

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Abbreviations

MS:

Murashige and Skoog medium

NAA:

α-Naphthaleneacetic acid

PCR:

Polymerase chain reaction

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Acknowledgments

The financial support of Department of Biotechnology, Govt. of India, New Delhi is gratefully acknowledged.

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Author notes

  1. Kottackal Poulose Martin

    Present address: Department of Plant Sciences, University of California, One Shields Ave, Davis, CA, 95616, USA

Authors and Affiliations

  1. Department of Biotechnology, University of Calicut, Calicut, 673 635, Kerala, India

    Awadhesh Kumar Arya, Kottackal Poulose Martin & Joseph Madassery

  2. Faculty of Biology, Institute of Botany, 11000, Belgrade, Serbia

    Aneta Sabovljevic

  3. Department of Medicine, IMS, Banaras Hindu University, Varanasi, 221005, India

    Awadhesh Kumar Arya

Authors
  1. Awadhesh Kumar Arya
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  2. Kottackal Poulose Martin
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  3. Aneta Sabovljevic
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  4. Joseph Madassery
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Corresponding authors

Correspondence to Kottackal Poulose Martin or Joseph Madassery.

Additional information

Communicated by J. Sadowski.

A. K. Arya and K. P. Martin contributed equally.

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Arya, A.K., Martin, K.P., Sabovljevic, A. et al. Transformation through agroinfection on decapitated shoot apex of field-growing Phyllanthus amarus . Acta Physiol Plant 33, 2011–2017 (2011). https://doi.org/10.1007/s11738-011-0716-x

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  • DOI: https://doi.org/10.1007/s11738-011-0716-x

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