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Asymbiotic seed germination and in vitro seedling development of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid

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Abstract

Continuing loss of native orchid habitat has lead to an increased emphasis on orchid conservation. Major obstacles in the production of native orchid seedlings for use in conservation have been: (1) development of efficient and reliable seed germination protocols and (2) an understanding of early seedling growth and development. Effects of six asymbiotic media (Modified Lucke, Murashige & Skoog, Lindemann, Vacin & Went, Malmgren Modified, Knudson C), four exogenous cytokinins (BA, Zea, Kin, 2-iP), and three photoperiods (0/24, 16/8, 24/0 h L/D) were examined on seed germination and early protocorm development of Habenaria macroceratitis, a rare native Florida terrestrial orchid. Finally, the effects of three photoperiods (8/16, 12/12, 16/8 h L/D) on in vitro seedling development were examined. Percent seed germination was highest on both LM and KC after seven weeks culture (LM = 89.1%, KC = 89.2%); however, protocorm development was enhanced on MM after both seven and 16 weeks. Both zeatin and kinetin at 1 µM enhanced seed germination (Zea = 58.1%, Kin = 47.2%). Final percent seed germination (91.7%) and protocorm development (Stage 4) was increased in the absence of light (0/24 h L/D). In␣vitro seedlings cultured under 8/16 h L/D conditions produced the highest number of tubers per seedling (1.06) with the greatest tuber (42.7 µg) and shoot (fwt = 69.5 µg) biomass and tuber diameter (3.1 mm).

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Abbreviations

2-iP:

6-(γ, γ-dimethylallylamino) purine

BA:

benzyladenine

Kin:

kinetin

KC:

Knudson C

LD:

long day

LN:

Lindemann’s

ML:

Modified Lucke’s

MM:

Malmgren’s Modified Terrestrial Orchid Medium

MS:

Murashige & Skoog

SD:

short day

VW:

Vacin & Went

Zea:

zeatin

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Acknowledgements

The authors thank Philip Kauth and Tim Johnson (Environmental Horticulture Department, University of Florida) for their reviews of this manuscript. Gratitude is conveyed to the San Diego County Orchid Society and the Florida Panther National Wildlife Refuge—US Fish and Wildlife Service for providing financial support of this project. Brand names are provided for references, the authors do not solely endorse these particular products.

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

    Scott L. Stewart & Michael E. Kane

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  1. Scott L. Stewart
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  2. Michael E. Kane
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Correspondence to Scott L. Stewart.

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Stewart, S.L., Kane, M.E. Asymbiotic seed germination and in vitro seedling development of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tiss Organ Cult 86, 147–158 (2006). https://doi.org/10.1007/s11240-006-9098-y

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  • DOI: https://doi.org/10.1007/s11240-006-9098-y

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