Establishment of encapsulation-dehydration technique for in vitro fragmented explants of Rosa hybrida L. cv. Helmut Schmidt

Abstract

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.

Integration of Cryopreservation and Tissue Culture for Germplasm Conservation and Propagation of Rosa pomifera ‘Karpatia’

“…Pawłowska and Bach (2011) achieved better results using the same method to cryopreserve meristems of Rosa 'New Dawn'. The encapsulation-dehydration technique was also described for the in vitro fragmented explants of R. hybrida 'Helmut Schmidt' using TTC (2,3,5-triphenyl-tetrazoliumchloride) to assess the survival (Mubbarakh et al, 2014).…”

In vitro PROPAGATION OF BANANA SHOOT TIPS AND CRYOPRESERVATION BY VITRIFICATION

“…"Over the last ten years, different studies and research are going on for the development of different efficient methods for cryopreservation. Recently, two important methods i.e., the vitrification" [10], Sakai et al 1991a; [11] and "encapsulation-dehydration techniques [12,13] have been developed for the cryopreservation of many plant species" (Engelmann and Takagi 2000; Engelmann et al [14]; Gonzalez-Arnao and Engelmann [15]; Sakai [16]; Sakai 1997).…”