Journal of Taiwan Agricultural Research

Author:Yih-Juh Shiau* and Guan-Wen Chen

Abstract:

    Strawberry (Fragaria × ananassa Duch.) is a high economic value perennial crop. It belongs to the Rosaceae family and is distributed in more than 70 countries. Therefore, the research on long-term storage of germplasm is important. The major objective of this study was to establish an optimum cryopreservation method of non-cold hardening Fragaria × ananassa Duch. cv. ‘Taoyuan No.1’. The results obtained on pretreatment of explants prior to plunge into liquid nitrogen (LN) were as follows: (1) The survival rate of shoot tips (1-2 mm³) was 100% when explants precultured on MS basal medium supplemented with 0.3 M sucrose for 14 d. (2) To increase sucrose levels from 0.6 M to 1.6 M in loading solution (LS) and then osmoprotected for 90 min, the survival rate of shoot tips remained 100%. (3) Survival rate of shoot tips achieved 96.7% ± 4.7% when treated with Plant Vitrification Solution 2 (PVS2) dehydration for 60 min. This percentage was significantly higher than treatments of PVS2 dehydration for 180 min (60.0% ± 14.1%) and 240 min (46.7% ± 17.0%). After PVS2 dehydration treatment, 10 shoot tip beads with 0.3 mL fresh PVS2 were transferred to a 2 mL cryotube and then plunged directly into LN for at least 1 h. After storage in LN, cryotubes were rapidly rewarmed in a water bath at 38℃ for 30 s. The final average regeneration rate of shoot tip beads (80%) was achieved after subcultured for 14 d. These research results appear promising for establishing and simplifying cryopreservation procedure of strawberry.

Key words:Fragaria × ananassa Duch., Cryopreservation, Loading solution, Osmoprotection, Plant Vitrification Solution 2

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