Conserving the Critically Endangered Anacamptis coriophora L. in Türkiye through Ex Vitro Seed Germination

Öz

Anacamptis coriophora (Orchidaceae) is a highly endangered orchid in Türkiye due to its excessive collection and the continuing deterioration of its habitat. In this study, the cultivation conditions of A. coriophora were determined. A sterile soil mixture was filled into jars and the fungal isolate (previously isolated from A. coriophora roots), Ceratobasidiaceae MG762693 was inoculated in separate glass jars, producing fungal compost when hyphae were developed. This fungal compost was then filled into pots where A. coriophora seed packs (0.001 g) were placed and subsequently moistened with sterile liquid nutrient medium. After 45 days of germination, fifty seedlings of approximately equal size were transferred directly to a natural environment and after 6 months of development the measuring of the tubers was done. The phenological process was then monitored until flowering. After 45 days, germination and developmental stages rates were determined from the seed packs in the pots inoculated with the Ceratobasidiaceae MG762693 fungal isolate and 64.3% germination and 11.75% leaf-rooted seedlings (stage 4) occurred. Plants flowered in June the following year, and the seeds ripened in July. The largest tuber in adult individuals was about 3 times the weight of first-year tubers. Each individual formed 2 or 3 tubers, thus increasing the number of tubers approximately 2.5 times in 2 years. In this study, ex vitro symbiotic seedlings were planted in the natural environment and a small population was formed in a 2-year period. The results revealed that orchids can be grown on a large scale with this method, both economically and for conservation and reintroduction.

Anahtar Kelimeler

• Orchidaceae
• Orchid cultivation
• Seed germination
• Symbiotic

Kaynakça

1. Aewsakul N, Maneesorn D, Serivichyaswat P, Taluengiit AT, Nontachaiyapoom S. 2013. Ex vitro symbiotic seed germination of Spathoglottis plicata Blume on common orchid cultivation substrates. Sci Horticultur, 160: 238–242.
2. Bektaş E, Cüce M, Sökmen A. 2013. In vitro germination protocorm formation and plantlet development of Orchis coriophora (Orchidaceae) a naturally growing orchid species in Turkey. Turkish J Botany, 37(2): 336-342. 10.3906/bot-1205-28.
3. Clements MA, Muir H, Cribb PJ. 1986. A Preliminary report on the symbiotic Germination of European terrestrial orchids. Kew Bullet, 41: 437–445.
4. Coats DJ, Dixon KW. 2007. Current perspectives in plant conservation biology. Aust J Bot, 55: 187-93. Çığ A, Demirer DE, İşler S. 2018. In vitro symbiotic germination potentials of some Anacamptis Dactylorhiza Orchis and Ophrys terrestrial orchid species. Appl Ecol Environ Res, 16(4): 5141-5155.
5. Deniz IG, Kömpe YÖ, Harzli I, Aytar EC, Mutlu VA, Uysal Dİ. 2022. From seed to flowering tuberous orchid using ex vitro symbiotic seed germination: A breakthrough study with Anacamptis sancta. Rhizosphere, 24: 100597. https://doi.org/10.1016/j.rhisph.2022.100597.
6. Eşitgen A, Ercişli S, Eken C. 2005. Effects of Mycorrhiza Isolates on Symbiotic Germination of Terrestrial Orchids (Orchis palustris Jacq. and Serapias vomeracea subsp. vomeracea (Burm.f.) Briq. in Turkey. Symbiosis, 38: 59-68.
7. quFatahi M, Vafaee Y, Nazari F, Tahir N.A. 2022. In vitro asymbiotic germination protocorm formation and plantlet development of Orchis simia Lam.: Athreatened terrestrial orchid species. South African J Botany, 151: 156-165.
8. Ghorbani A, Gravendeel B, Naghibi F de Boer H. 2014. Wild orchid tuber collection in Iran: a wake-up call for conservation. Biodiver Conserv, 23: 2749–2760.

9. Govaerts R, Bernet P, Kratochvil K, Gerlach G, Carr G, Alrich P, Pridgeon AM, Pfahl J, Campacci MA, Baptista DH, Tigges H, Shaw J, Cribb P, George A, Kreuz K, Wood JJ. 2017. World checklist of Orchidaceae. Facilitated by the Royal Botanic Gardens Kew, URL: https://www.scienceopen.com/document?vid=2e89a8a9-1cc4-49b5-b8fc-1a54fa2f9f48 (accessed date: April 16, 2023).
10. Gümüş C, Ellialtıoğlu Ş. 2012. Seed Germination and Development of Serapias vomeracea (Burm.fil.) Briq. ssp. orientalis Greuter in Tissue Culture. Res J Biotechnol, 7(3): 4-8.
11. Hopper SD. 2000. How well do phylogenetic studies inform the conservation of Australian plants? Aust J Bot, 48: 321-28.
12. Joppa LN, Roberts DL, Myers N, Pimm SL. 2011a. Biodiversity hotspots house most undiscovered plant species. Proceed National Acad Sci USA, 108: 13171–13176.
13. Joppa LN, Roberts DL, Pimm SL. 2011b. How many species of flowering plants are there? Proceed Royal Soc B, 278: 554–559.
14. Khasim SM, Hedge SN, Arnao MTG, Thammasiri K. 2020. Orchid biology: Recent Trends and hallenges. Springer, New York, USA, pp: 547.
15. Kömpe YÖ, Mutlu VA. 2021. Ex vitro symbiotic germination of the seeds of Anacamptis coriophora (L.) R.M. Bateman Pritgeon and M.W. Chase and Orchis anatolica Boiss. Biol Futur, 72: 509-516. https://doi.org/10.1007/s42977-021-00100-5.
16. Kömpe YÖ, Karakaya H, Mutlu VA. 2021. Symbiotic propagation of the wet meadow orchid Anacamptis laxiflora for conservation and reintroduction. Fresenius Environ Bullet, 30(9): 10535-10544.
17. Kömpe YÖ, Mutlu VA, Özkoç İ, Demiray S, Bozkurt S. 2022. Fungal diversity and ex vitro symbiotic germination of Serapias vomeracea (Orchidaceae). Acta Botan Croatica, 81(1): 108-116. DOI: 10.37427/botcro-2022-008.
18. Kreziou A, de Boer H, Gravendeel B. 2016. Harvesting of salep orchids in North-western Greece continues to threaten natural populations. Oryx, 50(3): 393–396. https:// doi. org/ 10. 1017/S0030 60531 50002 65.
19. León-Yánez S, Valencia R, Pitman N, Endara L, Ulloa Ulloa C, Navarette H. 2011. Red book of the endemic plants of Ecuador. Quito: Pontificia Universidad Católica del Ecuador, Quito, Ecuador, pp: 957.
20. Löki V, Tökölyi J, Süveges K, Lovas-Kiss A, Hürkan K, Sramko G, Molnar AV. 2015. The orchid flora of Turkish graveyards: a comprehensive field survey. Willdenowia, 45(2): 231-243.
21. Mutlu, V. A., & Kömpe, Y. O. (2020). Mycorrhizal fungi of some Orchis species of Turkey. Pakistan Journal of Botany, 52(2), 687-695.
22. Oktalira FT, Whitehead MR, Linde CC. 2019. Mycorrhizal specificity in widespread and narrow-range distributed Caladenia orchid species. Fungal Ecol, 42: 100869.
23. Pujasatria GC, Miura C, Kaminaka H. 2020. In vitro symbiotic germination: A revitalized heuristic approach for orchid species conservation. Plants, 9(12): 1742.
24. Qin H, Yang Y, Dong S. 2017. Threatened species list of China’s higher plants. Biodiver Sci, 25: 696–744.
25. Quay L, McComb JA, Dixon KW. 1995. Methods for ex vitro germination of Australian terrestrial orchids. Hortsci, 30: 1445-1446. https://doi.org/10.21273/HORTSCI.30.7.1445
26. Sezik E. 2002. Turkish orchids and salep. Acta Pharm Turcica, 44: 151–157
27. Sosa V, Platas T. 1998. Extinction and persistence of rare orchids in Veracruz Mexico. – Conservation Biology 12: 451-455.
28. Steinfort U, Verdugo G, Besoain X, Cisternas MA. 2010. Mycorrhizal association and symbiotic germination of the terrestrial orchid Bipinnula fimbriata (Poepp.) Johnst (Orchidaceae). Flora-Morphol Distrib Funct Ecol Plants, 205: 811–817.
29. Wang H, Fang H, Wang Y, Duan L, Guo S. 2011. In situ seed baiting techniques in Dendrobium officinale Kimuraet Migo and Dendrobium nobile Lindl.: the endangered Chinese endemic Dendrobium (Orchidaceae). World J Microbiol Biotechnol, 27: 2051–2059.
30. Wu K, Zeng S, Lin D, Da Silva T, Bu Z, Zhang J, Duan J. 2014. In vitro propagation and reintroduction of the endangered Renanthera imschootiana Rolfe. PLoS ONE, 9: e110033. doi: 10.1371/J.pone.0110033.
31. Zhang YY, Wu KL, Zhang JX, Deng RF, Duan J, Da Silva T, Huang WC, Zeng SJ. 2015. Embryo development in association with asymbiotic seed germination in vitro Paphiopedilum armeniacum S.C. Chen et F.Y. Liu. Scie Report, 5: 16356.
32. Zhang S, Yang Y, Li J, Qin J, Zhang W, Huang W, Hu H. 2018. Physiological diversity mpeof orchids. Plant Divers, 40(4): 196–208. https://doi.org/10.1016/j.pld.2018.06.003.
33. Zhang Y, Li YY, Chen XM, Guo SX, Li YI. 2020. Efect of diferent mycobionts on symbiotic germination and seedling growth of Dendrobium offcinale an important medicinal orchid. Botan Stud, 61: 2.