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http://dx.doi.org/10.7732/kjpr.2019.32.6.689

Cryopreservation of in vitro Grown Shoot Tips of Strawberry (Fragaria × ananassa Duch.) Genetic Resources by Droplet-vitrification  

Lee, Young-Yi (National Agrobiodiversity Center, NAS, RDA)
Balaraju, Kotnala (National Agrobiodiversity Center, NAS, RDA)
Song, Jae-Young (National Agrobiodiversity Center, NAS, RDA)
Yi, Jung-Yoon (National Agrobiodiversity Center, NAS, RDA)
Lee, Sun-Yi (Department of Horticultural Crop Research, NIHHS, RDA)
Lee, Jung-Ro (National Agrobiodiversity Center, NAS, RDA)
Yoon, Munsup (National Agrobiodiversity Center, NAS, RDA)
Kim, Haeng-Hoon (Department of Well-being Resources, Sunchon National University)
Publication Information
Korean Journal of Plant Resources / v.32, no.6, 2019 , pp. 689-697 More about this Journal
Abstract
This study describes an efficient and widely applicable droplet-vitrification following cryopreservation for shoot tips of strawberry (Fragaria × ananassa Duch.) cvs. 'Wonkyo3114' and 'Gurumi40'. The shoot tips were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.5M). Precultured explants were osmoprotected with loading solution (LS, C4) containing 20% glycerol and 20% sucrose for 40 min and exposed to dehydration solution (B5) containing 40% glycerol and 40% sucrose for 40 min at 25℃, Subsequently, the explants were transferred onto droplets containing 2.5 μL PVS3 on sterilized aluminum foils (4 cm × 0.5 cm) prior to direct immersion in liquid nitrogen (LN) for 1 h. The highest regrowth rate (%) in both the cultivars was obtained when the shoot tips were precultured with MS + 0.3M sucrose for 40 h at 25℃. The cryopreserved shoots tips exhibited 55% regrowth rate by culturing in NH4NO3-free MS medium supplemented with 3% sucrose, 1.0 g/L casein, 1.0 mg/L GA3, and 0.5 mg/L BA for 5 weeks and in MS medium supplemented with 0.5 mg/L GA3 for 8 weeks. This result shows that droplet-vitrification could be employed as a promising method for cryostorage of strawberry germplasm.
Keywords
Cryopreservation; Dehydration; Regrowth; Strawberry; Shoot tips;
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