CRYOPRESERVATION OF TISSUE-CULTURED SHOOT PRIMORDIA FROM SHOOT APICES OF CULTURED PROTOCORMS IN VANDA PUMILA FOLLOWING ABA PRECULTURE AND DESICCATION

摘要

A cryopreservation protocol including preculture with ABA followed by desiccation was developed for tissue-cultured shoot primordia of Vanda pumila (Orchidaceae). After a 3-day-preculture period in B5 liquid medium supplemented with 1.0 mg/l ABA, shoot primordia were desiccated under 40-45% relative humidity down to similar to 24% relative water content, then cryopreserved in liquid nitrogen (LN). After 10 days of recovery in B5 liquid medium supplemented with 0.02 mg/l BAP, cryopreserved shoot primordia showed a survival rate of 65.0 +/- 7.5%, The regrown shoot primordia from cryopreservation displayed no abnormalities in chromosome number or cell structure, and could induce new meristematic tissues like those of the non-cryopreserved controls. [References: 19]