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Acta Physiologiae Plantarum

, Volume 35, Issue 4, pp 1373–1379 | Cite as

Cryopreservation of pharmaceutically important orchid Dendrobium chrysanthum Wall. ex Lindl. using vitrification based method

  • P. Mohanty
  • M. C. DasEmail author
  • S. Kumaria
  • P. Tandon
Short Communication

Abstract

Our present study constitutes the successful and efficient protocol for cryopreservation of Dendrobium chrysanthum. D. chrysanthum Wall. ex Lindl. is a pharmaceutically valuable, ornamental epiphytic orchid of temperate and subtropical regions. On account of excellent herbal medicinal value and horticultural importance, D. chrysanthum is becoming rare due to over exploitation. For long-term conservation of this orchid, protocorm-like bodies of D. chrysanthum were excised and used for cryopreservation by encapsulation–vitrification. In this cryogenic procedure, PLBs were initially osmoprotected with a mixture of 0.4 M sucrose and 2 M glycerol, incorporated in the encapsulation matrix (comprising of 3 % (w/v) sodium alginate and 0.1 M CaCl2). Encapsulated protocorm-like bodies (PLBs) were then precultured on MS liquid medium supplemented with different concentrations of sucrose (0.06, 0.3, 0.5, 0.7 M), and loaded in a loading solution (comprised of 2 M glycerol and 0.4 M sucrose) for different duration to make the precultured PLBs tolerant to plant vitrification solution 2 (PVS2). Subsequently, the PLBs were subjected to PVS2 (Sakai et al. 1990) treatment at different time of exposure (minutes) and temperatures (0 °C and 25 °C). Encapsulated–vitrified PLBs were plunged directly into liquid nitrogen and stored for 1 h. Optimum result (survival 63.2 % and regrowth 59.9 %) was obtained when the beads treated with loading solution for 80 min followed by PVS2 treatment for 100 min. Regenerated plants showed normal morphology as that of control plants.

Keywords

Dendrobium chrysanthum Protocorm-like bodies A-naphthaleneacetic acid 6-benzylaminopurine Plant vitrification solution 2 

Abbreviations

EV

Encapsulation–vitrification

PVS2

Plant vitrification solution 2

PLB

Protocorm-like bodies

Notes

Acknowledgments

The first author is thankful to UGC Centre for Advanced Studies in Botany, North Eastern Hill University (NEHU), Shillong, India for financial support. SAIF, NEHU is highly acknowledged for supply of liquid nitrogen.

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Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2012

Authors and Affiliations

  1. 1.Plant Biotechnology LaboratoryCentre for Advanced Studies in Botany North Eastern Hill UniversityShillongIndia

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