Cryopreservation of orchid seeds through rapid and step

Ecuador has a great variety of climatic regions that potentiate biodiversity. The family constitutes one of the most important of the country, having Orchidaceae identified about 4032 species with a high degree of endemism, therefore the development and research of alternative methods of storage and conservation of species is a strategy of primary interest for researchers and for society in general. In cryopreservation, temperatures reach below -190°C in order to paralyze the chemical reactions and keep the plant material viable for long periods. The present research focuses on the development of protocols for cryopreservation of seeds, aimed at the preservation of biodiversity, focusing on the family , for the subsequent generation of a seed bank. The Orchidaceae assays were performed on seeds of , Epidendrum quitensium, Sobralia rosea and . Two freezing rates were tested: rapid freezing at Epidendrum anderssonii -196°C; and step freezing at -22°C, -60°C to 196°C, further analyzed four combinations from Dimethylsulfoxide DMSO, glycerol and sucrose (DMSO 1M; DMSO 1M + glycerol 1M; DMSO 1M + sucrose 1M; DMSO 1M + glycerol 0,5M + sucrose 0,5M). The best results were obtained both in rapid and stepped freezing without the use of cryo-protective substances, by introducing the seeds directly into liquid nitrogen. Species of the genus presented Epidendrum a more efficient response in comparison to . The viability of the seeds Sobralia was evaluated by the tetrazolium test. Bence Mátyás ( ) Corresponding author: [email protected] : Conceptualization, Investigation, Methodology, Writing – Original Draft Preparation; : Investigation, Author roles: Cerna M Valdivieso P Methodology; : Investigation, Methodology, Writing – Original Draft Preparation; : Project Administration, Writing – Original Draft Cella R Mátyás B Preparation, Writing – Review & Editing; : Investigation, Methodology, Writing – Original Draft Preparation Aucapiña C No competing interests were disclosed. Competing interests: Cerna M, Valdivieso P, Cella R How to cite this article: et al. Cryopreservation of orchid seeds through rapid and step freezing methods 2018, :209 (doi: ) [version 1; referees: awaiting peer review] F1000Research 7 10.12688/f1000research.13622.1 © 2018 Cerna M . This is an open access article distributed under the terms of the , which Copyright: et al Creative Commons Attribution Licence permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Data associated with the article are available under the terms of the (CC0 1.0 Public domain dedication). Creative Commons Zero "No rights reserved" data waiver The author(s) declared that no grants were involved in supporting this work. Grant information: 20 Feb 2018, :209 (doi: ) First published: 7 10.12688/f1000research.13622.1 1 2 3 4