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High-efficiency cryopreservation of the medicinal orchid Dendrobium nobile Lindl.

  • P. Mohanty
  • M. C. DasEmail author
  • S. Kumaria
  • P. Tandon
Original Paper

Abstract

An efficient protocol for cryopreservation of protocorm like bodies (PLBs) of Dendrobium nobile, based on encapsulation–dehydration (ED) and encapsulation–vitrification (EV), was established. In both cryogenic procedures, PLBs were initially osmoprotected with a mixture of 0.4 M sucrose and 2 M glycerol, incorporated in the encapsulation matrix [comprising 3% (w/v) sodium alginate and 0.1 M CaCl2]. Out of the two methods, EV resulted in higher survival (78.1%) and regrowth (75.9%) than ED (53.3 and 50.2% respectively). Incorporation of 0.4 M sucrose and 2 M glycerol in the encapsulation matrix resulted in higher survival percentage after cryopreservation. In both the cases (ED and EV), shoots regenerated from cryopreserved PLBs with an intermediary PLB formation. Regenerated shoots were successfully rooted in the medium containing 1.5 mg/l Indole-3 butyric acid. Successful acclimatization of plantlets was obtained in the compost containing brick pieces and charcoal chunks (1:1) + a top layer of moss with a maximum survivability (82%). EV method proved to be most appropriate way to cryopreserve the PLBs of D. nobile. Regenerated plantlets showed normal morphology as that of control plants.

Keywords

Dendrobium nobile Protocorm like bodies Cryopreservation 

Abbreviations

ED

Encapsulation–dehydration

EV

Encapsulation–vitrification

PVS2

Plant vitrification solution 2

PLB

Protocorm like bodies

NAA

α-Naphthaleneacetic acid

IBA

Indole-3 butyric acid

Notes

Acknowledgments

This research was financially supported by the Centre for Advanced studies in Botany, North-Eastern Hill University (NEHU), Shillong, India. The authors are also thankful to SAIF, NEHU for supply of liquid nitrogen.

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Plant Biotechnology Laboratory, Centre for Advanced Studies in BotanyNorth Eastern Hill UniversityShillongIndia

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