New methods to improve symbiotic propagation of temperate terrestrial orchid seedlings from axenic culture to soil
A. L. Batty A B D , M. C. Brundrett A C , K. W. Dixon A and K. Sivasithamparam BA Science Directorate, Botanic Gardens and Parks Authority, Kings Park and Botanic Garden, West Perth, WA 6005, Australia.
B School of Earth and Geographical Sciences (Soil Science Discipline), Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, WA 6009, Australia.
C School of Plant Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, WA 6009, Australia.
D Corresponding author. Email: abatty@bgpa.wa.gov.au
Australian Journal of Botany 54(4) 367-374 https://doi.org/10.1071/BT04023
Submitted: 12 February 2005 Accepted: 27 October 2005 Published: 22 June 2006
Abstract
This research aimed to improve the success of soil transfer of terrestrial orchid seedlings after symbiotic germination in the laboratory. Three native Western Australian terrestrial orchids (Caladenia arenicola Hopper & A.P.Brown, Diuris magnifica D.L.Jones and Thelymitra crinita Lindley) were used in this study. The key to improved seedling survival on transfer to soil was found to be the use of an intermediate stage between the Petri dish and soil where larger seedlings were grown in an axenic environment with controlled humidity. There was no apparent benefit of pre-inoculating potting medium with appropriate strains of mycorrhizal fungi for subsequent growth of symbiotic seedlings under glasshouse conditions. Initial survival of seedlings in the glasshouse was high. However, some seedlings failed to produce tubers (from modified roots or droppers) necessary for plant survival through the summer dormancy period, and this caused survival to decrease to 40–60% of the glasshouse-grown seedlings in the first year. The initiation of tubers on droppers by C. arenicola was inversely correlated with leaf size, with smaller plants more likely to form tubers. This suggests that leaves and tubers were competing for resources. However, larger seedlings that did tuberise had larger tubers that were more likely to survive summer dormancy. There was no correlation between leaf size and root tuber size in D. magnifica, but the number of tubers produced was greatest in larger plants. As with C. arenicola, plants of D. magnifica and T. crinita with larger tubers were more likely to survive summer dormancy. Methods developed in this study enable the production of both actively growing symbiotic seedlings and dormant tubers which improve the success of translocation of laboratory-grown terrestrial orchids to field sites.
Acknowledgments
The conservation of terrestrial orchids in Western Australia requires further research to help prevent loss of diversity. Western Power, the supplier of electricity for Perth, recognised the plight of the states threatened flora and provided the necessary financial support for this study to help save critically endangered terrestrial orchids. The first two authors were supported by funding from the ARC.
Anderson AB
(1991) Symbiotic and asymbiotic germination and growth of Spiranthes magnicamporum (Orchidaceae). Lindleyana 6, 183–186.
Batty AL,
Dixon KW,
Brundrett M, Sivasithamparam K
(2001a) The long-term storage of mycorrhizal fungi and seed as a tool for the conservation of endangered terrestrial orchids. Australian Journal of Botany 49, 619–628.
| Crossref | GoogleScholarGoogle Scholar |
Batty AL,
Dixon KW,
Brundrett M, Sivasithamparam K
(2001b) Constraints to symbiotic germination of terrestrial orchid seed in Mediterranean woodland. New Phytologist 152, 511–520.
| Crossref | GoogleScholarGoogle Scholar |
Batty AL,
Brundrett MC,
Dixon KW, Sivasithamparam K
(2006) In situ symbiotic seed germination and propagation of terrestrial orchid seedlings for establishment at field sites. Australian Journal of Botany 54, 375–381.
Clements MA, Ellyard RK
(1979) The symbiotic germination of Australian terrestrial orchids. American Orchid Society Bulletin 48, 810–816.
Clements MA,
Muir H, Cribb PJ
(1986) A preliminary report on the symbiotic germination of European terrestrial orchids. Kew Bulletin 41, 437–445.
Debeljak N,
Regvar M,
Dixon KW, Sivasithamparam K
(2002) Induction of tuberisation in vitro with jasmonic acid and sucrose in an Australian terrestrial orchid, Pterostylis sanguinea. Plant Growth Regulation 36, 253–260.
| Crossref | GoogleScholarGoogle Scholar |
Hahn EJ, Paek KY
(2001) High photosynthetic photon flux and high CO2 concentration under increased number of air exchanges promote growth and photosynthesis of four kinds of orchid plantlets in vitro. In Vitro Cellular and Developmental Biology. Plant 37, 678–682.
Hollick P,
Senaratna T,
McComb J,
Bunn E, Dixon K
(2002) Response to paclobutrazol of symbiotic mycorrhizal fungi and dropper (tuber stalk) formation of host seedlings. Plant Growth Regulation 36, 31–39.
| Crossref | GoogleScholarGoogle Scholar |
Masuhara G, Katsuya K
(1994) In situ and in vitro specificity between Rhizoctonia spp. and Spiranthes sinensis (Persoon.) Ames. var. amoena (M.Beiberstein) Hara (Orchidaceae). New Phytologist 127, 711–718.
| Crossref |
Perkins AJ, McGee PA
(1995) Distribution of the orchid mycorrhizal fungus Rhizoctonia solani, in relation to its host, Pterostylis acuminata, in the field. Australian Journal of Botany 43, 565–575.
| Crossref | GoogleScholarGoogle Scholar |
Peterson RL,
Uetake Y, Zelmer C
(1998) Fungal symbioses with orchid protocorms. Symbiosis 25, 29–55.
Pridgeon AM, Chase MW
(1995) Subterranean axes in three Diurideae (Orchidaceae): morphology, anatomy, and systematic significance. American Journal of Botany 82, 1473–1495.
| Crossref |
Ramsay MM
(1998) Re-establishment of the lady’s slipper orchid (Cypripedium calceolus L.) in Britain. Botanical Journal of the Linnean Society 126, 173–181.
| Crossref | GoogleScholarGoogle Scholar |
Ramsay RR,
Dixon KW, Sivasithamparam K
(1986) Patterns of infection and endophytes associated with Western Australian orchids. Lindleyana 1, 203–214.
Scade A,
Brundrett MC,
Batty AL,
Dixon KW, Sivasithamparam K
(2006) Survival of transplanted terrestrial orchid seedlings in urban bushland habitats with high or low weed cover. Australian Journal of Botany 54, 383–389.
Stewart SL, Zettler LW
(2002) Symbiotic germination of three semi-aquatic rein orchids (Habenaria repens, H. quinquiseta, H. macroceratitis) from Florida. Aquatic Botany 72, 25–35.
| Crossref | GoogleScholarGoogle Scholar |
Warcup JH
(1971) Specificity of mycorrhizal associations in some Australian terrestrial orchids. New Phytologist 70, 41–46.
| Crossref |
Warcup JH
(1973) Symbiotic germination of some Australian terrestrial orchids. New Phytologist 72, 387–392.
| Crossref |
Zelmer CD, Currah RS
(1997) Symbiotic germination of Spiranthes lacera (Orchidaceae) with naturally occurring endophyte. Lindleyana 12, 142–148.
Zettler LW
(1997) Terrestrial orchid conservation by symbiotic seed germination: techniques and perspectives. Selbyana 18, 188–194.
Zettler LW, McInnis TM
(1992) Propagation of Platanthera integrilabia (Correll) Luer, an endangered terrestrial orchid, through symbiotic seed germination. Lindleyana 7, 154–161.
Zettler LW, McInnis TM
(1993) Symbiotic seed germination and development of Spirathes cerua (L.) L.C.Rich. and Goodyera pubescens (Wild.) R.Br. (Orchidaceae: Spiranthoideae). Lindleyana 8, 155–162.
Zettler LW,
Barrington FV, McInnis TM
(1995) Developmental morphology of Spiranthes odorata seedlings in symbiotic culture. Lindleyana 10, 211–216.