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Development and Optimization of a Germination Assay and Long-Term Storage for Cannabis sativa Pollen

Pollen viability and storage is of great interest to cannabis breeders and researchers to maintain desirable germplasm for future use in breeding or for biotechnological and gene editing applications. Here, we report a simple and efficient cryopreservation method for long-term storage of Cannabis sa...

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Autores principales: Gaudet, Daniel, Yadav, Narendra Singh, Sorokin, Aleksei, Bilichak, Andriy, Kovalchuk, Igor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7284845/
https://www.ncbi.nlm.nih.gov/pubmed/32456251
http://dx.doi.org/10.3390/plants9050665
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author Gaudet, Daniel
Yadav, Narendra Singh
Sorokin, Aleksei
Bilichak, Andriy
Kovalchuk, Igor
author_facet Gaudet, Daniel
Yadav, Narendra Singh
Sorokin, Aleksei
Bilichak, Andriy
Kovalchuk, Igor
author_sort Gaudet, Daniel
collection PubMed
description Pollen viability and storage is of great interest to cannabis breeders and researchers to maintain desirable germplasm for future use in breeding or for biotechnological and gene editing applications. Here, we report a simple and efficient cryopreservation method for long-term storage of Cannabis sativa pollen. Additionally, the bicellular nature of cannabis pollen was identified using DAPI (4′,6-diamidino-2-phenylindole) staining. A pollen germination assay was developed to assess cannabis pollen viability and used to demonstrate that pollen collected from different principal growth stages exhibited differential longevity. Finally, a simple and efficient method that employs pollen combined with baked whole wheat flour and subsequent desiccation under vacuum was developed for the long-term cryopreservation of C. sativa pollen. Using this method, pollen viability was maintained in liquid nitrogen after four months, suggesting long-term preservation of cannabis pollen.
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spelling pubmed-72848452020-06-15 Development and Optimization of a Germination Assay and Long-Term Storage for Cannabis sativa Pollen Gaudet, Daniel Yadav, Narendra Singh Sorokin, Aleksei Bilichak, Andriy Kovalchuk, Igor Plants (Basel) Protocol Pollen viability and storage is of great interest to cannabis breeders and researchers to maintain desirable germplasm for future use in breeding or for biotechnological and gene editing applications. Here, we report a simple and efficient cryopreservation method for long-term storage of Cannabis sativa pollen. Additionally, the bicellular nature of cannabis pollen was identified using DAPI (4′,6-diamidino-2-phenylindole) staining. A pollen germination assay was developed to assess cannabis pollen viability and used to demonstrate that pollen collected from different principal growth stages exhibited differential longevity. Finally, a simple and efficient method that employs pollen combined with baked whole wheat flour and subsequent desiccation under vacuum was developed for the long-term cryopreservation of C. sativa pollen. Using this method, pollen viability was maintained in liquid nitrogen after four months, suggesting long-term preservation of cannabis pollen. MDPI 2020-05-23 /pmc/articles/PMC7284845/ /pubmed/32456251 http://dx.doi.org/10.3390/plants9050665 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Gaudet, Daniel
Yadav, Narendra Singh
Sorokin, Aleksei
Bilichak, Andriy
Kovalchuk, Igor
Development and Optimization of a Germination Assay and Long-Term Storage for Cannabis sativa Pollen
title Development and Optimization of a Germination Assay and Long-Term Storage for Cannabis sativa Pollen
title_full Development and Optimization of a Germination Assay and Long-Term Storage for Cannabis sativa Pollen
title_fullStr Development and Optimization of a Germination Assay and Long-Term Storage for Cannabis sativa Pollen
title_full_unstemmed Development and Optimization of a Germination Assay and Long-Term Storage for Cannabis sativa Pollen
title_short Development and Optimization of a Germination Assay and Long-Term Storage for Cannabis sativa Pollen
title_sort development and optimization of a germination assay and long-term storage for cannabis sativa pollen
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7284845/
https://www.ncbi.nlm.nih.gov/pubmed/32456251
http://dx.doi.org/10.3390/plants9050665
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