Novel Apoplastic Antifreeze Proteins of Deschampsia antarctica as Enhancer of Common Cell Freezing Media for Cryobanking of Genetic Resources, a Preliminary Study

Author:

Short Stefania E.1ORCID,Zamorano Mauricio1ORCID,Aranzaez-Ríos Cristian1,Lee-Estevez Manuel2,Díaz Rommy3ORCID,Quiñones John3ORCID,Ulloa-Rodríguez Patricio4ORCID,Villalobos Elías Figueroa5ORCID,Bravo León A.6ORCID,Graether Steffen P.7ORCID,Farías Jorge G.1ORCID

Affiliation:

1. Department of Chemical Engineering, Universidad de La Frontera, Av. Francisco Salazar 01145, P.O. Box 54D, Temuco 4811230, Chile

2. Faculty of Health Sciences, Universidad Autónoma de Chile, Av. Alemania 1090, Temuco 4810101, Chile

3. Faculty of Agricultural and Environmental Sciences, Universidad de La Frontera, Av. Francisco Salazar 01145, Temuco 4811230, Chile

4. Department of Agronomical Sciences, Universidad Católica del Maule, Av. Carmen 684, Curicó 3341695, Chile

5. Nucleus of Research in Food Production, Faculty of Natural Resources, Universidad Católica de Temuco, Manuel Montt 056, Temuco 4813302, Chile

6. Department of Agronomical Sciences and Natural Resources, Universidad de La Frontera, Av. Francisco Salazar 01145, Temuco 4811230, Chile

7. Department of Molecular and Cellular Biology, University of Guelph, 50 Stone Rd E, Guelph, ON N1G 2W1, Canada

Abstract

Antifreeze proteins (AFPs) are natural biomolecules found in cold-adapted organisms that lower the freezing point of water, allowing survival in icy conditions. These proteins have the potential to improve cryopreservation techniques by enhancing the quality of genetic material postthaw. Deschampsia antarctica, a freezing-tolerant plant, possesses AFPs and is a promising candidate for cryopreservation applications. In this study, we investigated the cryoprotective properties of AFPs from D. antarctica extracts on Atlantic salmon spermatozoa. Apoplastic extracts were used to determine ice recrystallization inhibition (IRI), thermal hysteresis (TH) activities and ice crystal morphology. Spermatozoa were cryopreserved using a standard cryoprotectant medium (C+) and three alternative media supplemented with apoplastic extracts. Flow cytometry was employed to measure plasma membrane integrity (PMI) and mitochondrial membrane potential (MMP) postthaw. Results showed that a low concentration of AFPs (0.05 mg/mL) provided significant IRI activity. Apoplastic extracts from D. antarctica demonstrated a cryoprotective effect on salmon spermatozoa, with PMI comparable to the standard medium. Moreover, samples treated with apoplastic extracts exhibited a higher percentage of cells with high MMP. These findings represent the first and preliminary report that suggests that AFPs derived from apoplastic extracts of D. antarctica have the potential to serve as cryoprotectants and could allow the development of novel freezing media.

Funder

Fondecyt

CONICYT National Doctorate Scholarships

NEXER

Publisher

MDPI AG

Subject

Molecular Biology,Biochemistry

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