Ascocenda Wangsa Gold is a new and fascinating orchid hybrid in the Malaysian flower industry. An efficient plant vitrification solution 2 (PVS2) cryopreservation technique was developed for protocorm-like bodies (PLBs) of Ascocenda Wangsa Gold orchid. Parameters assessed included the effect of PVS2 exposure periods, thawing duration, temperature, and culture conditions based on 2,3,5-triphenyltetrazolium chloride absorbance readings and regrowth rates. A regrowth rate of 33.3% was obtained after 2 months when the PLBs were dehydrated in PVS2 for 30 min. The growth rate was improved to 47% when thawing was conducted at 45 °C for 85 s. The highest growth rate (53.3%) was obtained when the PLBs were subjected to a 7-day dark treatment before being transferred to a 16-h/8-h light/dark photoperiod. Histological analyses were conducted to study the morphology of cryopreserved and noncryopreserved PLBs of Ascocenda Wangsa Gold. The vitrification protocol developed in this study is a feasible and safe method for strengthening the germplasm conservation of this orchid for commercial purposes.
This study was carried out to evaluate the encapsulation-dehydration technique on IFEs of Rosa hybrida L. cv. Helmut Schmidt. The survival of IFEs was first assessed based on the effects of four sucrose concentrations (0, 0.25, 0.5 and 0.75 M) at different durations (0, 24 and 48 hours). The IFEs were then encapsulated and osmoprotected on shaker at 110 rpm with various sucrose concentrations (0, 0.1, 0.5 and 0.75 M). Subsequently, encapsulated IFEs were dehydrated under laminar air flow during three different time periods (0, 3 and 6 hours) on oven sterilized 50 g silica gel. The encapsulated IFEs were plunged into liquid nitrogen for a minimum duration of 24 hour. Encapsulated IFEs were thawed at 40ºC for 90 seconds and cultured on fullstrength MS medium supplemented with 3% sucrose for a week in dark, a week in semi-light and an additional 1 week under full light exposure. IFEs were then evaluated for survival by 2,3,5-Triphenyltetrazolium Chloride assay at 490 nm. The best conditions for the encapsulation-dehydration of IFEs of Rosa hybrida L. cv. Helmut Schmidt was obtained when IFEs were precultured in 0.25 M sucrose for 24 hours, osmoprotected in 0.75 M sucrose and dehydrated for 3 hours. Histological analysis showed cryopreserved IFEs with intensely stained nucleus but with severely damaged membranes.
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