Cryopreservation of Teucrium polium L. shoot-tips by vitrification and encapsulation-dehydration

Abstract

Teucrium polium L. with the common name of Felty Germander is one of the plants flora that is widely used in folk medicine in many Middle East countries, it is an endangered plant species and must be highly considered for preservation. Cryopreservation of T. polium by vitrification and encapsulation-dehydration was successfully achieved in this study. Shoot-tips were excised aseptically from in vitro grown plants and incubated for 3 days on solid hormone free-Murashige and Skoog (HF-MS) media supplemented with 0.3 M sucrose under complete darkness at 24 ± 1 °C. In vitrification, shoot-tips were loaded in 0.4 M sucrose and 2 M glycerol for 20 min followed by desiccation with different combinations and concentrations of plant vittrification solution 2 (PVS2), before immersion in Liquid Nitrogen (LN). Whereas for the encapsulation-dehydration; shoot-tips were encapsulated in calcium alginate and dehydrated under laminar air flow cabinet for 0, 3, 6, or 9 h. A total of 60 % of the cryopreserved vitrified shoot-tips survived when desiccated in concentrated PVS2 solution for 20 min, whereas, 28 % of the cryopreserved vitrified shoot-tips were regrown after 20 min of desiccation by two step increase in PVS2 concentration. Complete survival were obtained for the non-cryopreserved encapsulated shoot-tips treated for 3 days in 0.5 M sucrose with MS media without or with 3 h of dehydration, whereas, only 20 % of the cryopreserved encapsulated shoot-tips were regrown. The procedures developed in this study are easy to handle and produced a high levels of shoot formation.