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, 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.
Highlights
Cannabis or hemp (Cannabis sativa L.) is an annual, primarily dioecious flowering plant
ToTo obtain a representation of the germination profile, a time-lapse of a pollen germination assay obtain a representation of the germination profile, a time-lapse of a pollen germination assay (PGA)
Pollen cryopreservation has been employed in a variety of agriculturally and medicinally important plant species for the preservation of elite germplasm
Summary
Cannabis or hemp (Cannabis sativa L.) is an annual, primarily dioecious flowering plant. Breeding projects may wish to store pollen for extended periods of time, where high value genetic material may be stored for future use or for biotechnological and gene editing applications that requires a quick and effective method for determining pollen viability [3,4,5]. Fluorescent stains such as fluorescein diacetate (FDA). Pollen germination rates were assessed period when stored at 4 ◦ C from males at different stages of floral development.
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