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Acta Physiologiae Plantarum

, Volume 35, Issue 9, pp 2813–2820 | Cite as

In vitro propagation and assessment of clonal fidelity of Nepenthes khasiana Hook. f.: a medicinal insectivorous plant of India

  • Soibam Purnima Devi
  • Suman Kumaria
  • Satyawada Rama Rao
  • Pramod Tandon
Original Paper

Abstract

An efficient in vitro protocol for large-scale multiplication of Nepenthes khasiana, a threatened insectivorous plant of India, has been developed from nodal stem segments. The highest shoot proliferation of 19.16 ± 0.23 shoots/explant was recorded in half-strength Murashige and Skoog (MS) medium supplemented with 2.5 mg/l kinetin, 2.0 mg/l 6-benzyl aminopurine, 3 % sucrose and 0.8 % agar. The best rooting was achieved in half-strength MS medium supplemented with 2.0 mg/l α-naphthalene acetic acid with an average of 9.04 ± 0.46 roots/shoot. The plantlets were successfully transferred to the greenhouse with survival rate of 92 %, exhibiting normal development. Cytological and random amplified polymorphic DNA (RAPD) analyses were carried out to assess the genetic integrity of the regenerated plantlets. Cytological analysis revealed no change in chromosome number with cells studied showing 2n = 80. Of the 80 primers screened for RAPD analysis, 14 primers resulted in clear and scorable bands. A total of 72 amplification products were obtained out of which only 4.1 % bands were polymorphic. Cluster analysis of the RAPD profile revealed an average similarity coefficient ranging from 0.98 to 1.0, thus suggesting genetic stability in the micropropagated plants of N. khasiana.

Keywords

Nepenthes khasiana Multiple shoots Nodal segments Genetic stability RAPD analysis 

Notes

Acknowledgments

The study was supported by Research Grant No. BT/04/NE/2009 from Department of Biotechnology, Government of India, New Delhi, India.

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Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2013

Authors and Affiliations

  • Soibam Purnima Devi
    • 1
  • Suman Kumaria
    • 1
  • Satyawada Rama Rao
    • 2
  • Pramod Tandon
    • 1
  1. 1.Department of BotanyNorth-Eastern Hill UniversityShillongIndia
  2. 2.Department of Biotechnology and BioinformaticsNorth-Eastern Hill UniversityShillongIndia

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