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In vitro cloning of Azadirachta indica from root explants

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Biologia Plantarum

Abstract

In vitro cultures of Azadirachta indica A. Juss. were raised by first culturing the root segments on modified Murashige and Skoog (MS) medium supplemented with 8.88 μM 6-benzylaminopurine (BAP), 9.84 μM N6-(2-isopentenyl) adenine (2iP), 5.71 μM indole-3-acetic acid (IAA), 81.43 μM adenine hemisulphate and 2.27 μM putrescine for 2 d followed by their transfer to the same medium except containing one-tenth of the initially used concentrations of BAP, 2iP and IAA. The regenerated shoots sustained proliferation in the basal medium supplemented with 1.11 μM BAP, 1.43 μM IAA and 135.72 μM adenine hemisulphate. The isolated shoots were rooted to produce plantlets in the presence of 2.46 μM indole-3-butyric acid (IBA). The plantlets showed uniform luxuriant growth under field conditions. True-to-type nature of the field-grown root-regenerated plants was ascertained by random amplified polymorphic DNA (RAPD) analysis.

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Abbreviations

BAP:

6-benzylaminopurine

2,4-D:

2,4-dichlorophenoxyacetic acid

IAA:

indole-3-acetic acid

IBA:

indole-3-butyric acid

IPA:

indole-3-propionic acid

2iP:

N6-(2-isopentenyl) adenine

MS medium:

Murashige and Skoog medium

NAA:

α-naphthaleneacetic acid

NOA:

α-naphthoxyacetic acid

PVP:

polyvinylpyrollidone

RAPD:

random amplified polymorphic DNA

TDZ:

thidiazuron

Z:

zeatin

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Acknowledgements

The authors thank the Director, National Botanical Research Institute (NBRI), Lucknow for the facilities provided. The first author also thanks the Council of Scientific and Industrial Research (CSIR), India, for awarding her Senior Research Fellowship. Thanks are also due to Mr. R.S. Tripathi and Mr. O.P. Sharma for technical assistance.

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Authors and Affiliations

  1. Tissue Culture Laboratory, National Botanical Research Institute, Lucknow, 226001, India

    K. Arora, M. Sharma & A. K. Sharma

  2. Plant Molecular Biology (Genomics) Division, National Botanical Research Institute, Lucknow, 226001, India

    J. Srivastava & S. A. Ranade

Authors
  1. K. Arora
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  2. M. Sharma
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  3. J. Srivastava
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  4. S. A. Ranade
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  5. A. K. Sharma
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Correspondence to A. K. Sharma.

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Arora, K., Sharma, M., Srivastava, J. et al. In vitro cloning of Azadirachta indica from root explants. Biol Plant 55, 164–168 (2011). https://doi.org/10.1007/s10535-011-0023-9

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  • DOI: https://doi.org/10.1007/s10535-011-0023-9

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