クランベリーおよびブルーベリー培養体茎頂の超低温保存

概要

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The objective of this study was to establish a cryopreservation protocol for shoot apices of cranberry (Vaccinium macrocarpon Ait. cv. Early Black) and highbush blueberryn (V. corybossum L. cv. Rancoccas). Cryopreservation was carried out via encapsulation-dehydration on shoot apices excised from in vitro cultures. In the encapsulation dehydration method of both species, the protocol leading to optimal regrowth was as follows: After cold-acclimation at 5℃ in the dark for 2 weeks, excised shoot tips were pretreated for 24hr at 25℃ on hormone-free culture medium with 0.4mol/L sucrose, then encapsulated and precultured in liquid culture medium with 0.8mol/L sucrose for 16hr. Precultured beads were dehydrated for 360-mins in the dessicator containing 50g silica gel at 18% moisture content (fresh weight basis) before cryostorage for 1hr. In addition, the effect of adding glycerol and/or salicylic acid to and loading solution was examined to enhance regrowth after cryopreservation. In the present study, it was shown that adding both 0.5mol/L (cranberry) or 1.0mol/L glycerol (highbush blueberry) and 50μmol/L resulted in high regrowth.
低温生物工学会の論文
2010-03-15