COLD HARDENING VS ABA AS A PRETREATMENT FOR MERISTEM CRYOPRESERVATION

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  • 1 National Clonal Germplasm Repository, 33447 Peoria Road, Corvallis, OR 97333

Cold hardening is an effective method for conditioning meristems for cryopreservation. ABA plays a role in hardening and produces increased hardiness in suspension cultured cells. This study was designed to determine if growth, in vitro, on ABA (5×10-5 M) for one week, would substitute for one week of cold hardening, and if ABA would provide additional conditioning when added in combination with cold hardening treatments. In vitro plantlets of Rubus spp. were grown for one week with or without cold hardening and with or without ABA. Meristems from these plants were frozen at 0.8C* min-1 to -35 C, then plunged into LN2, thawed, and plated on recovery medium. One month after thawing, cold-hardened plants with and without ABA treatment had recovery rates of up to 83%. Survival of plants grown at room temperature ranged from zero to 8% and zero to 28% for plants grown on ABA at room temperature. At the rates tested, ABA is less effective than cold hardening in conditioning apical meristems of in vitro Rubus plants for cryopreservation and provides no additional protection to cold-hardened meristems.

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