Long-term conservation of dormant buds of Prunus dulcis (Miller) DA Webb. using three different new cryotechniques

Choudhary, R.; Malik, S. K.; Chaudhury, R.; Sharma, K. C.

Romanian Biotechnological Letters 19(4): 9575-9584


ISSN/ISBN: 1224-5984
Accession: 066288828

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Winter dormant buds of Prunus dulcis were cryopreserved using either two-step freezing, encapsulation-dehydration or vitrification. A two-step freezing cryoprotocol preceded by desiccation to 18 to 28% moisture content was developed. Recovery conditions, including dark incubation and rehydration in sterile moist moss grass for different durations, led to higher survival. For encapsulation-dehydration, alginate beads containing descaled bud were dehydrated for 1-3 days in various sucrose concentrations (0.3, 0.5, 0.75 or 1.0 M). Bead desiccation was performed using laminar air flow for either 1-6 h. Treatment of alginate beads with 0.75 M sucrose was more effective in promoting re-growth of explants after immersion in liquid nitrogen. For vitrification, descaled buds were directly immersed for 20, 40, 60, 90 or 120 min in a vitrification solution (PVS2). Re-growth of explants was also observed following vitrification and this reached 50% with increasing duration of the PVS2 treatment from 20 to 90 min. Overall, the highest frequency of explant re-growth was obtained when explants were subjected to two step freezing followed by encapsulation-dehydration in the presence of 0.75 M along with 24 h sucrose dehydration pre-treatment and followed by desiccation for 5 h in laminar air flow.