@conference{
author = "Trajković, Milena and Antonić Reljin, Dragana and Trailović, Maja and Trifunović-Momčilov, Milana and Subotić, Angelina and Jevremović, Slađana",
year = "2018",
abstract = "Cryopreservation represents a suitable method for long term storage of different plant genetic
resources. The aim of this study was to develop protocol for cryopreservation of Viola cornuta
shoot tips using one step freezing method with chemical dehydration of tissue with modified
Plant Vitrification Solutions (PVS2 or PVS3). Shoot tips (1-2 mm) of two-week cold acclimated
shoots were cultured on ½MS medium with 0.3 M sucrose for one day before treatment with loading
solution (2 M glycerol, 0.4 M sucrose) for 30 min. Osmotic dehydration with PVS2 solution (30% glycerol, 15% ethylene glycol and 15% DMSO in liquid ½MS medium with 0.4 M sucrose) were
tested at 0 °C or 24 °C. Osmotic dehydration with PVS3 (50% sucrose, 50% glycerol in liquid ½MS
medium) were tested at 24 °C for 45 min. After the treatment the explants were directly immersed
in liquid nitrogen (LN) for at least one day. Re-warming was performed at 42 °C in water bath
for 2 min. After re-warming, the PVS solutions were replaced with unloading solution containing
1.2 M sucrose for 20 min. Re-warmed shoot tips were cultured on ½MS medium with 0.1 mg L-1
BAP. We observed that PVS2 solution is cytotoxic for V. cornuta shoot tips and cannot be used for
cryopreservation. However, cryopreservation with PVS3 solution was successful, where 71.9-100%
shoot tips survived treatment before immersion to LN and 31-40% survived after re-warming from
LN. Regrowth of cryopreserved shoot tips with new well-formed leaves was obtained after four
weeks of culture.",
publisher = "Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Faculty of Biology, University of Belgrade",
journal = "Book of abstracts. 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia",
title = "Cryopreservation of Viola cornuta shoot tips using vitrification procedure",
pages = "91-92",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6722"
}