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ARS Home » Plains Area » Fargo, North Dakota » Edward T. Schafer Agricultural Research Center » Weed and Insect Biology Research » Research » Publications at this Location » Publication #410738
Research Project: Enhancing Pollinator Health and Availability Through Conservation of Genetic Diversity and Development of Novel Management Tools and Strategies

Location: Weed and Insect Biology Research

Title: Monarch butterfly conservation through male germplasm cryopreservation

Author
item Grula, Courtney
item Rajamohan, Arun
item Rinehart, Joe

Submitted to: Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/21/2024
Publication Date: 6/28/2024
Citation: Grula, C.C., Rajamohan, A., Rinehart, J.P. 2024. Monarch butterfly conservation through male germplasm cryopreservation. Science. https://doi.org/10.1038/s41598-024-65624-x.
DOI: https://doi.org/10.1038/s41598-024-65624-x

Interpretive Summary: Monarch butterfly populations are in decline in North America. Monarchs have been recently classified as endangered by the International Union for Conservation of Nature (ICUN), leading to increased conservation efforts. One method of conservation is cryopreservation. Cryopreservation is a method that preserves biological material by cooling to very low temperatures with the potential for long term storage. Our objective was to develop a cryopreservation protocol for monarch butterfly sperm to ensure effective, extended storage in liquid nitrogen. In this study sperm from male monarch butterflies was cryopreserved in a cryoprotective medium and stored in liquid nitrogen. To assess sperm viability after cryopreservation, a subsample of preserved sperm was thawed, and the viability was qualified and compared to non-cryopreserved sperm samples. Through cryopreservation, our aim was to preserve and store both genetic material and viable sperm. Our results show that cryopreservation does not impact the survival of monarch sperm. This marks the first documented success in the cryopreservation of monarch genetic material, laying the groundwork for continued cryopreservation and conservation efforts in this species.

Technical Abstract: Monarch butterfly, Danaus plexippus (L.), populations have declined, and there has been increased interest in conservation efforts. Our approach to conservation is through cryopreservation of germinal cells. Our goal was to develop a cryopreservation protocol for monarch spermatozoa for successful long-term storage. Cryopreservation would serve as a reserve for monarch germplasm, which could be utilized in the event of population loss. The sperm cells collected from monarch butterflies were cryopreserved and stored in liquid nitrogen. Post-cryopreserved sperm was thawed, and the viability was qualified. Through cryopreservation, we hope to preserve and store genetic material and viable sperm. To date, this is the first report of successful cryopreservation of monarch germplasm which sets the foundation for cryostorage and may be extensible to other vulnerable lepidopterans.