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Development and optimization of a germination assay and long-term storage for Cannabis sativa pollen

Daniel Gaudet, View ORCID ProfileNarendra Singh Yadav, Aleksei Sorokin, Andrii Bilichak, Igor Kovalchuk
doi: https://doi.org/10.1101/2020.03.19.999367
Daniel Gaudet
1Department of Biological Sciences, University of Lethbridge, Lethbridge T1K 3M4, Alberta, Canada
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Narendra Singh Yadav
1Department of Biological Sciences, University of Lethbridge, Lethbridge T1K 3M4, Alberta, Canada
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  • ORCID record for Narendra Singh Yadav
Aleksei Sorokin
1Department of Biological Sciences, University of Lethbridge, Lethbridge T1K 3M4, Alberta, Canada
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Andrii Bilichak
1Department of Biological Sciences, University of Lethbridge, Lethbridge T1K 3M4, Alberta, Canada
2Morden Research and Development Center, Agriculture and Agri-Food Canada, Morden, MB, Canada
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Igor Kovalchuk
1Department of Biological Sciences, University of Lethbridge, Lethbridge T1K 3M4, Alberta, Canada
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  • For correspondence: igor.kovalchuk@uleth.ca
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Abstract

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, we have deciphered the bicellular nature of cannabis pollen using DAPI staining. We have also standardized a pollen germination assay to assess the viability of cannabis pollen, and found pollen collected from different principal growth stages exhibits different longevity. Finally, we developed a long-term storage method which includes pollen combination with baked whole wheat flower and desiccation under vacuum for cryopreservation. By using this method, we were able to maintain germination viability in liquid nitrogen after 4 months, suggesting potentially indefinite preservation of cannabis pollen.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted March 20, 2020.
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Development and optimization of a germination assay and long-term storage for Cannabis sativa pollen
Daniel Gaudet, Narendra Singh Yadav, Aleksei Sorokin, Andrii Bilichak, Igor Kovalchuk
bioRxiv 2020.03.19.999367; doi: https://doi.org/10.1101/2020.03.19.999367
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Development and optimization of a germination assay and long-term storage for Cannabis sativa pollen
Daniel Gaudet, Narendra Singh Yadav, Aleksei Sorokin, Andrii Bilichak, Igor Kovalchuk
bioRxiv 2020.03.19.999367; doi: https://doi.org/10.1101/2020.03.19.999367

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