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Plant Regeneration Through Callus Organogenesis and True-to-Type Conformity of Plants by RAPD Analysis in Desmodium gangeticum (Linn.) DC.

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Abstract

An efficient plant regeneration protocol was established for an endangered ethnomedicinal plant Desmodium gangeticum (Linn.) DC. Morphogenic calli were produced from 96 % of the cultures comprising the immature leaf explants on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (4.0 mg l−1) in combination with 6-benzylaminopurine (BA; 0.8 mg l−1). For callus regeneration, various concentrations of BA (1.0–5.0 mg l−1) or thidiazuron (TDZ; 1.0–5.0 mg l−1) alone or in combination with indole-3-acetic acid (IAA; 0.2–1.0 mg l−1) were used. Highest response of shoot regeneration was observed on MS medium fortified with TDZ (4.0 mg l−1) and IAA (0.5 mg l−1) combination. Here, 100 % cultures responded with an average number of 22.3 shoots per gram calli. Inclusion of indole-3-butyric acid in half MS medium favored rooting of recovered shoots. Out of 45 rooted plants transferred to soil, 40 survived. Total DNA was extracted from the leaves of the acclimatized plants of D. gangeticum. Analysis of random amplified polymorphic DNA using 13 arbitrary decanucleotide primers showed the genetic homogeneity in all the ten plants regenerated from callus with parental plant, suggesting that shoot regeneration from callus could be used for the true-to-type multiplication of this plant.

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Abbreviations

BA:

N 6-Benzylaminopurine

2,4-D:

2,4-Dichlorophenoxyacetic acid

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

MS:

Murashige and Skoog (1962)

NAA:

1-Naphthalene acetic acid

RAPD:

Random amplified polymorphic DNA

TDZ:

Thidiazuron

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Acknowledgments

TDT acknowledges the financial assistance from UGC in the form of a major research project (project no. 38-233/2009).

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  1. Postgraduate and Research Department of Botany, St. Thomas College, Pala, Arunapuram (P.O.), 686574, Kottayam (Dt.), Kerala, India

    Meena K. Cheruvathur, Jyothi Abraham & T. Dennis Thomas

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  1. Meena K. Cheruvathur
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Correspondence to T. Dennis Thomas.

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Cheruvathur, M.K., Abraham, J. & Thomas, T.D. Plant Regeneration Through Callus Organogenesis and True-to-Type Conformity of Plants by RAPD Analysis in Desmodium gangeticum (Linn.) DC.. Appl Biochem Biotechnol 169, 1799–1810 (2013). https://doi.org/10.1007/s12010-013-0117-2

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