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Physiology and Molecular Biology of Plants

, Volume 18, Issue 2, pp 169–176 | Cite as

Micropropagation of Pithecellobium dulce (Roxb.) Benth—a multipurpose leguminous tree and assessment of genetic fidelity of micropropagated plants using molecular markers

  • Pooja Goyal
  • Sumita Kachhwaha
  • S. L. Kothari
Research Article

Abstract

An efficient and reproducible protocol has been developed for in vitro propagation of Pithecellobium dulce (Roxb.) Benth (a multipurpose leguminous tree) from field grown nodal segments (axillary bud). Shoot bud induction occurred from nodal explants of 15-years-old tree on Murashige and Skoog (MS) basal medium supplemented with 4.4 μM 6-benzyladenine (BA) and multiplication was achieved on MS medium supplemented with 4.4 μM BA + 0.73 μM phenylacetic acid (PAA) i.e. up to 7 shoot buds in the period of 5–6 weeks. Addition of adenine sulphate (AdS) to this medium further enhanced the number of shoot buds up to 10. Proliferating shoot cultures were established by repeatedly subculturing primary culture on fresh medium (MS + 4.4 μM BA + 0.73 μM PAA) after every 25 days. In vitro rooting was achieved on MS medium supplemented with 2.46 μM Indole-3-butyric acid (IBA) + 41.63 μM activated charcoal (AC). The micropropagated shoots with well developed roots were acclimatized in green house in pots containing sand, soil and manure (1:1:1). Genetic stability of micropropagated clones was evaluated using Random amplified polymorphic DNA (RAPD) and Inter simple sequence repeat (ISSR) markers. The amplification products were monomorphic in micropropagated plants and similar to those of mother plant. No polymorphism was detected revealing the genetic uniformity of micropropagated plants. This is the first report of an efficient protocol for regeneration of P. dulce through organogenesis, which can be used for further genetic transformation and pharmaceutical purposes.

Keywords

Legume tree Nodal segments Activated charcoal Adenine sulphate RAPD, ISSR marker 

Abbreviations

AC

Activated charcoal

AdS

Adenine sulphate

BA

6-benzyladenine

IAA

Indole-3-acetic acid

IBA

Indole-3-butyric acid

ISSR

Inter-simple sequence repeats

Mo

Mother plant

MS

Murashige and Skoog

NAA

α-naphthaleneacetic acid

PAA

Phenylacetic acid

RAPD

Random amplified polymorphic DNA

WPM

Woody plant medium

Notes

Acknowledgements

Pooja Goyal thanks Council for Scientific and Industrial Research (CSIR), New Delhi for the award of SRF. Dr. Arunima Sinha thanks University Grants Comission (UGC) for financial support in the form of R&D project 33-154/2007(SR).

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Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2012

Authors and Affiliations

  1. 1.Centre for Converging Technologies (CCT)University of RajasthanJaipurIndia
  2. 2.Department of BotanyUniversity of RajasthanJaipurIndia

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