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Asymbiotic and symbiotic seed germination of Eulophia alta (Orchidaceae)—preliminary evidence for the symbiotic culture advantage

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Abstract

Eulophia alta (Linnaeus) Fawcett & Rendle seeds collected from the Florida Panther National Wildlife Refuge (Collier County, FL; FPNWR) were used in a screen of five asymbiotic orchid seed germination media to determine their effectiveness in promoting germination and protocorm development. In addition, 10 fungal isolates collected from the roots of E. alta at sites in the FPNWR, Highlands County (FL), and Goethe State Forest (Levy County, FL; GSF), and a fungal isolate from the roots of Spiranthes brevilabris collected from GSF were screened for their effectiveness at promoting in vitro symbiotic germination of E. alta seeds. After 18 weeks asymbiotic culture, seeds sown on PhytoTechnology Orchid Seed Sowing Medium germinated to a higher percentage (87.9%) and had a higher percentage of protocorms with developing protomeristems (32.7%) than seeds cultured on Knudson C, Malmgren Modified Terrestrial Orchid Medium, ½-strength Murashige & Skoog, or Vacin & Went Modified Orchid Medium. Significantly more leaf-bearing protocorms were observed on PhytoTechnology Orchid Seed Sowing Medium (0.8%) and Vacin & Went Modified Orchid Medium (1.3%) than other media tested. Of the fungi tested, one fungal isolate (Ealt-396) promoted germination to 69.0%, two isolates promoted germination to less than 0.75% and did not support further protocorm development, and eight isolates did not support germination. Seeds co-cultured in darkness with Ealt-396 grew more rapidly than asymbiotic seedlings following germination. In addition, co-cultured (=symbiotic) seedlings continued to develop more rapidly than asymbiotic seedlings upon transfer to 16/8 h light/dark photoperiod. Symbiotic seed culture of E. alta may be a more desirable method of propagation since protocorms develop more rapidly than seeds sown on asymbiotic media. Symbiotic seedlings may be more appropriate for reintroduction to natural areas than asymbiotic seedlings since symbiotic seedlings could serve to inoculate soils with a germination promoting mycobiont.

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Abbreviations

1/5-PDA:

1/5-strength Potato dextrose agar

½MS:

½-strength Murashige & Skoog

CMA:

Corn meal agar

FPNWR:

Florida Panther National Wildlife Refuge

dd:

Distilled deionized

KC:

Knudson C

L/D:

Light/dark

MM:

Malmgren Modified Terrestrial Orchid Medium

P723:

PhytoTechnology Orchid Seed Sowing Medium

TZ:

Tetrazolium

VW:

Vacin & Went Modified Orchid Medium

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Acknowledgements

The authors thank the Florida Panther National Wildlife Refuge—US Fish and Wildlife Service for providing financial and logistical support for this project. Appreciation is also extended to Dr. Carrie Reinhardt Adams (University of Florida) for the use of microscopic equipment, and Dr. James Kimbrough (University of Florida) for assistance in fungal identification. The authors would also like to thank Philip Kauth (University of Florida) and Nancy Philman (University of Florida) for help constructing and revising this paper. Brand names are provided for references; the authors do not solely endorse these particular products.

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Authors and Affiliations

  1. Department of Environmental Horticulture, University of Florida, PO Box 110675, Gainesville, FL, 32611, USA

    Timothy R. Johnson, Scott L. Stewart, Daniela Dutra & Michael E. Kane

  2. Florida Panther National Wildlife Refuge, U.S. Fish and Wildlife Service, 3860 Tollgate Blvd., Suite 300, Naples, FL, 34114, USA

    Larry Richardson

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  1. Timothy R. Johnson
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  2. Scott L. Stewart
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  3. Daniela Dutra
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Correspondence to Timothy R. Johnson.

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Johnson, T.R., Stewart, S.L., Dutra, D. et al. Asymbiotic and symbiotic seed germination of Eulophia alta (Orchidaceae)—preliminary evidence for the symbiotic culture advantage. Plant Cell Tiss Organ Cult 90, 313–323 (2007). https://doi.org/10.1007/s11240-007-9270-z

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