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Application of encapsulation-vitrification in combination with air dehydration enhances cryotolerance of Chrysanthemum morifolium shoots tips

Jeon, Su Min, Arun, Muthukrishnan, Lee, Seung-Yeol, Kim, Chang Kil
Scientia horticulturae 2015 v.194 pp. 91-99
Chrysanthemum morifolium, air drying, air flow, carbon, cryopreservation, cultivars, fructose, germplasm, glucose, maltose, nitrogen, regrowth, shoot tips, sucrose, survival rate, vitrification, water content
In this study, we report an efficient encapsulation-vitrification-dehydration protocol for cryopreservation of Chrysanthemum morifolium shoot tips. Various factors influencing cryopreservation such as explant size (1.0–2.0mm), carbon source (0.0–0.9M sucrose, glucose, fructose and maltose), duration of pre-culture (0–3 days), duration of treatment with loading solution (0–300min), duration of treatment with plant vitrification solution (0–120min with PVS2 or PVS3), and duration of air-dehydration (0–6h) were investigated. Shoot tips of 1.0mm size that were encapsulated, and pre-cultured for 1 day in medium enriched with 0.6M sucrose, treated with loading solution for 180min, dehydrated with PVS3 for 60min, and sequentially air dried for 4h (33.0% water content) in laminar air flow cabinet prior to exposure with liquid nitrogen (LN) for 1h was found to be the most suitable regime for cryopreservation of Chrysanthemum shoot tips. Under optimal conditions, the highest survival rate of 91.3%, and shoot regrowth rate of 80.3% was obtained using cultivar ‘Borami’. The method developed was successfully adapted to three more cultivars, ‘Secret pink’, ‘Yellow cap’, and ‘White ND’ which showed a survival rate of 60.3–87.3%, and shoot regrowth rate of 55.3–70.0%. To our knowledge this could be the first report in cryopreservation combining the advantage of encapsulation-vitrification and encapsulation-dehydration. We believe that the developed protocol will be a tool to conserve Chrysanthemum germplasm.