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ABSTRACT
Diospyros kaki Thunb. was divided into four types based on astringency in the fruit at harvest, presence of seed, and flesh colour: The factors that affected cryopreservation of D. kaki Thunb. were studied by modified droplet vitrification: preculture condition and PVS2 treatment time. After cryopreservation, different concentration of Pluronic F-68 in the regenerating medium was also investigated. A suitable protocol was established: shoot tips of axillaries were precultured on MS (1/2) medium with 0.5mol/L sucrose and 10g/L Pluronic F-68 for 5 days, which were loaded in loading solution for 20 min at 20°C, and incubated in PVS2 for 1.5 h at 0°C prior to direct plunge into liquid nitrogen. After rapid thawing at 40°C warm water bath and rinsed, the shoot tips were plated on the regeneration medium supplemented with 1.0mg/L TDZ, 0.6g/L PVP, 0.1% Pluronic F-68, 30g/L sucrose and 7g/L agar for one week in dark prior to exposure to normal conditions. Using this procedure in cryopreservation of D. kaki Thunb., the best regeneration rate was 80%, the average rate was 73% and the difference of four types was not significant at P=0.05.