title: Cryopreservation of shoot tips of Picrorhiza kurroa Royle ex Benth, an indigenous endangered medicinal plant, through vitrification.
abstract: The cryopreservation of shoot tips of Picrorhiza kurroa Royle ex Benth (IC 266698), an endangered medicinal plant of India was investigated. Shoot tips (about 1 mm in length) excised from four-week-old proliferating shoot cultures were precultured on MS medium supplemented with various osmotica before dehydrating with PVS2 solution at 0 degrees C. The dehydrated shoot tips were directly immersed in LN2. Following cryopreservation, and after rapid rewarming at 45 degrees C, shoot tips were quickly washed with 1.2 M sucrose solution and then plated on solidified shoot culture medium. Shoot tips were successfully cryopreserved by vitrification, when they were precultured on medium supplemented with 5% DMSO at 4 degrees C for two days before dehydrating in PVS2 for 10-20 minutes at 0 degrees C. Average survival in terms of normal shoot formation after 4 wks of plating was about 20% without callus formation. Cold hardening of shoot cultures for four weeks at 4 degrees C significantly improved the survival and shoot regeneration of cryopreserved shoot tips to 70% and 35%, respectively.
authors.affiliation: Tissue Culture and Cryopreservation Unit, National Bureau of Plant Genetic Resources, Pusa Campus, New Delhi-110012, India. neelam@nbpgr.delhi.nic.in
authors.forename: Neelam
authors.initials: N
authors.lastname: Sharma
authors.forename: Bindu
authors.initials: B
authors.lastname: Sharma
mesh_terms: D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D003470:Culture Media; D046508:Culture Techniques; D004121:Dimethyl Sulfoxide; D007194:India; D032261:Picrorhiza; D018514:Plant Structures; D010946:Plants, Medicinal; D014022:Tissue Survival
chemical_list: D003451:Cryoprotective Agents; D003470:Culture Media; D004121:Dimethyl Sulfoxide