Abstract
Helonias bullata, commonly known as swamp pink, is a perennial rhizomatous herb native to the eastern coast of the United States that is found mainly in forested wetlands, swamps, and mountain bogs. Habitat destruction has caused H. bullata to become rare, and it is federally listed as threatened. Protocols were developed for in vitro germination, shoot micropropagation, shoot establishment in soil, and seed cryopreservation that will assist in the safeguarding and conservation of dwindling natural populations. Seeds were germinated in vitro on plant growth regulator (PGR)-free 1/3-strength Murashige and Skoog (MS) medium after seed sterilization in H2O2 and used for establishment of shoot cultures. Shoot multiplication and elongation occurred on 1/3-strength MS medium containing 0.25–1 mg/l kinetin or 1–2 mg/l trans-zeatin followed by growth on PGR-free medium. All shoots (100%) rooted when planted directly into potting media. Rapid-immersion seed cryopreservation resulted in statistically significant increases in germination in vitro or in potting mix compared to control treatments. H. bullata seeds share characteristics of both orthodox and recalcitrant seeds including rapid loss of viability over time, survival at 5°C, and survival after cryopreservation. Therefore, they are most accurately classified as intermediate-type seed.
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Acknowledgements
We thank Bill Brumback and the New England Wildflower Society and Tom Patrick from the Georgia Department of Natural Resources for providing seeds. The authors gratefully acknowledge the help of Kylie Bucalo for student training, laboratory infrastructure maintenance, and media preparation. We also thank the Georgia Institute of Technology for providing materials and supplies for undergraduate research in support of this project and the Greenville Zoo Conservation Fund for an award to help carry out the research.
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Perullo, N., Determann, R.O., Cruse-Sanders, J.M. et al. Seed cryopreservation and micropropagation of the critically endangered species swamp pink (Helonias bullata L.). In Vitro Cell.Dev.Biol.-Plant 51, 284–293 (2015). https://doi.org/10.1007/s11627-015-9679-8
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DOI: https://doi.org/10.1007/s11627-015-9679-8