Cryobiotechnological Studies in Vanilla: The Orchid of Multi-industrial Uses

In this work we provide new information about several cryobiotechnological studies conducted using Vanilla planifolia species as a case study. We evaluated the effect of three vitrification-based techniques, droplet-vitrification (D-V), V cryo-plate (V-Cp) and D cryo-plate (D-Cp), on the survival of two types of in vitro explants (shoot-tips and root-tips). Using differential scanning calorimetry (DSC), there were defined thermal characteristics of shoot-tips subjected to several desiccation times with SG following D-Cp procedure. It was calculated the removal of osmotically active water (OAW) by the effect of desiccation duration after the osmoprotective steps of preconditioning and loading. Using ISSR markers with six selected ISSR primers, we also evaluated the impact of tissue culture and of cryopreservation on genetic stability of vanilla shoot-tips dehydrated with the vitrification solution PVS2 and comparing D-V and V-Cp procedures. All these investigations are allowing the development and optimization of reliable cryopreservation protocols for long-term storage of Vanilla germplasm.