Preculture Condition for Stable Recovery of Strawberry (Fragaria × ananassa Duch.) Shoot Tips after Cryopreservation Using Droplet Vitrification
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This study describes an efficient and stable droplet vitrification following cryopreservation of strawberry shoottip (Fragaria × ananassa Duch.) accessions ‘Massey’ and ‘MDUS3816’. The shoot tips were precultured in Murashige andSkoog (MS) liquid medium supplemented with sucrose (0.3-0.7M). Precultured explants were osmoprotected with loadingsolution (LS, C4) containing 17.5% glycerol and 17.5% sucrose for 40 min and exposed to dehydration solution (B1)containing 50% glycerol and 50% sucrose for 40 min at 25oC. Subsequently, the explants were transferred onto dropletscontaining 2.5 µL PVS3 on sterilized aluminum foils (4 ㎝× 0.5 ㎝) prior to direct immersion in liquid nitrogen (LN) for 1h. The highest regrowth rate (%) in both the cultivars was obtained when the shoot tips were precultured with 0.3M sucrosefor 30 h + 0.5M sucrose for 16 h at 25oC. The cryopreserved shoots tips exhibited 57.8 % recovery rate by culturing inNH4NO3-free MS medium supplemented with 3% sucrose, 1.0 g/L casein, 1.0㎎/L GA3, and 0.5 ㎎/L BA for 5 weeks andin MS medium supplemented with 0.5 ㎎/L GA3 for 8 weeks. Variation was not observed in both of ploidy analysis andmorphological investigation on plantlets of two accessions cryopreserved under variable preculture conditions.
Abstract
This study describes an efficient and stable droplet vitrification following cryopreservation of strawberry shoottip (Fragaria × ananassa Duch.) accessions ‘Massey’ and ‘MDUS3816’. The shoot tips were precultured in Murashige andSkoog (MS) liquid medium supplemented with sucrose (0.3-0.7M). Precultured explants were osmoprotected with loadingsolution (LS, C4) containing 17.5% glycerol and 17.5% sucrose for 40 min and exposed to dehydration solution (B1)containing 50% glycerol and 50% sucrose for 40 min at 25oC. Subsequently, the explants were transferred onto dropletscontaining 2.5 µL PVS3 on sterilized aluminum foils (4 ㎝× 0.5 ㎝) prior to direct immersion in liquid nitrogen (LN) for 1h. The highest regrowth rate (%) in both the cultivars was obtained when the shoot tips were precultured with 0.3M sucrosefor 30 h + 0.5M sucrose for 16 h at 25oC. The cryopreserved shoots tips exhibited 57.8 % recovery rate by culturing inNH4NO3-free MS medium supplemented with 3% sucrose, 1.0 g/L casein, 1.0㎎/L GA3, and 0.5 ㎎/L BA for 5 weeks andin MS medium supplemented with 0.5 ㎎/L GA3 for 8 weeks. Variation was not observed in both of ploidy analysis andmorphological investigation on plantlets of two accessions cryopreserved under variable preculture conditions.
참고문헌 (23)
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