Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources

Sweet potato (Ipomoea batatas) is one of the ten most important staple crops and provides a livelihood for many people around the globe. To adapt to ever-changing circumstances farmers and breeders need to have access to a broad diversity of germplasm. This study focuses on the development of a cryo...

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Autores principales: Wilms, Hannes, Fanega Sleziak, Natalia, Auweraer, Maarten van der, Brands, Martijn, Verleije, Matthijs, Hardeman, Dirk, André, Edwige, Panis, Bartholomeus
Formato: Journal Article
Lenguaje:Inglés
Publicado: Springer 2020
Materias:
Acceso en línea:https://hdl.handle.net/10568/109851
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author Wilms, Hannes
Fanega Sleziak, Natalia
Auweraer, Maarten van der
Brands, Martijn
Verleije, Matthijs
Hardeman, Dirk
André, Edwige
Panis, Bartholomeus
author_browse André, Edwige
Auweraer, Maarten van der
Brands, Martijn
Fanega Sleziak, Natalia
Hardeman, Dirk
Panis, Bartholomeus
Verleije, Matthijs
Wilms, Hannes
author_facet Wilms, Hannes
Fanega Sleziak, Natalia
Auweraer, Maarten van der
Brands, Martijn
Verleije, Matthijs
Hardeman, Dirk
André, Edwige
Panis, Bartholomeus
author_sort Wilms, Hannes
collection Repository of Agricultural Research Outputs (CGSpace)
description Sweet potato (Ipomoea batatas) is one of the ten most important staple crops and provides a livelihood for many people around the globe. To adapt to ever-changing circumstances farmers and breeders need to have access to a broad diversity of germplasm. This study focuses on the development of a cryopreservation protocol that allows the long term storage of different sweet potato cultivars. For this, a droplet vitrification protocol was optimized, comparing several parameters; preculture method (0.3 M sucrose vs no preculture); meristem position (axillary vs apical); plant age (3 to 9 weeks); regeneration medium (MS + 2.22 µM BA, Hirai and MS); and length of loading solution treatment (20 to 360 min). Two months after cryopreservation, the regeneration rates of the meristems were compared, which resulted in significant differences for the preculture method, meristem position and loading solution. With these new insights an optimized droplet vitrification protocol was developed with the following parameters: use of 3–9 week old axillary meristems, no preculture phase, 20 min LS treatment, 30 min PVS2 treatment, exposure to liquid nitrogen by droplet vitrification, warming treatment in RS for 15 min, 1 day 0.3 M sucrose recuperation culture, 1 month MS + 2.22 µM BA followed by 1 month of MS cultures. This protocol was subsequently tested on 10 representative accessions resulting in a post cryopreservation regeneration rate of more than 40% for 70% of the tested cultivars, showing that this protocol could be implemented for a large portion of existing sweet potato collections.
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spelling CGSpace1098512025-11-12T05:39:17Z Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources Wilms, Hannes Fanega Sleziak, Natalia Auweraer, Maarten van der Brands, Martijn Verleije, Matthijs Hardeman, Dirk André, Edwige Panis, Bartholomeus ipomea batatas cryopreservation germplasm conservation research methods criopreservación conservación del germoplasma investigación Sweet potato (Ipomoea batatas) is one of the ten most important staple crops and provides a livelihood for many people around the globe. To adapt to ever-changing circumstances farmers and breeders need to have access to a broad diversity of germplasm. This study focuses on the development of a cryopreservation protocol that allows the long term storage of different sweet potato cultivars. For this, a droplet vitrification protocol was optimized, comparing several parameters; preculture method (0.3 M sucrose vs no preculture); meristem position (axillary vs apical); plant age (3 to 9 weeks); regeneration medium (MS + 2.22 µM BA, Hirai and MS); and length of loading solution treatment (20 to 360 min). Two months after cryopreservation, the regeneration rates of the meristems were compared, which resulted in significant differences for the preculture method, meristem position and loading solution. With these new insights an optimized droplet vitrification protocol was developed with the following parameters: use of 3–9 week old axillary meristems, no preculture phase, 20 min LS treatment, 30 min PVS2 treatment, exposure to liquid nitrogen by droplet vitrification, warming treatment in RS for 15 min, 1 day 0.3 M sucrose recuperation culture, 1 month MS + 2.22 µM BA followed by 1 month of MS cultures. This protocol was subsequently tested on 10 representative accessions resulting in a post cryopreservation regeneration rate of more than 40% for 70% of the tested cultivars, showing that this protocol could be implemented for a large portion of existing sweet potato collections. 2020-10 2020-10-15T09:01:05Z 2020-10-15T09:01:05Z Journal Article https://hdl.handle.net/10568/109851 en Open Access application/pdf Springer Wilms, H.; Fanega Sleziak, N.; Van der Auweraer, M.; Brands, M.; Verleije, M.; Hardeman, D.; Andre, E.; Panis, B. (2020) Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources. Scientific Reports 10: 14674 ISSN: 2045-2322
spellingShingle ipomea batatas
cryopreservation
germplasm conservation
research
methods
criopreservación
conservación del germoplasma
investigación
Wilms, Hannes
Fanega Sleziak, Natalia
Auweraer, Maarten van der
Brands, Martijn
Verleije, Matthijs
Hardeman, Dirk
André, Edwige
Panis, Bartholomeus
Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources
title Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources
title_full Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources
title_fullStr Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources
title_full_unstemmed Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources
title_short Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources
title_sort development of a fast and user friendly cryopreservation protocol for sweet potato genetic resources
topic ipomea batatas
cryopreservation
germplasm conservation
research
methods
criopreservación
conservación del germoplasma
investigación
url https://hdl.handle.net/10568/109851
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