Mycorrhiza

, Volume 24, Issue 1, pp 35–43 | Cite as

Symbiotic seed germination and protocorm development of Aa achalensis Schltr., a terrestrial orchid endemic from Argentina

  • Fracchia Sebastián
  • Silvani Vanesa
  • Flachsland Eduardo
  • Terada Graciela
  • Sede Silvana
Original Paper

Abstract

Aa achalensis is an endangered terrestrial orchid endemic from Argentina. In vitro symbiotic seed germination was evaluated for its propagation. Five different fungal strains were isolated from this species: two Rhizoctonia-like related to Thanatephorus cucumeris and three ascomicetaceous fungi belonging to Phialophora graminicola and one to an uncultured Pezizaceae. All five isolates promoted seed germination being one T. cucumeris strain the most effective. After 16 weeks of growth, 30 % of A. achalensis protocorms developed until seedlings with two/four leaves in this treatment. These findings open an opportunity to the knowledge and preservation of this species.

Keywords

Orchid mycorrhiza Seed germination Dark septate endophytes Conservation 

References

  1. 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. Aust J Bot 54:375–381CrossRefGoogle Scholar
  2. Bianco C, Cantero JJ (1985) Las especies de Orchidaceae del suroeste de la provincia de Cordoba. Revista de la Universidad Nacional de Rio Cuarto 5:131–141Google Scholar
  3. Bidartondo MI, Read DJ (2008) Fungal specificity bottlenecks during orchid germination and development. Mol Ecol 17:3707–3716PubMedGoogle Scholar
  4. Bidartondo M, Burghardt B, Gebauer G, Bruns TD, Read DJ (2004) Changing partners in the dark: isotopic and molecular evidence of ectomycorrhizal liaisons between forest orchids and trees. Proc R Soc Lond 271:1799–1806CrossRefGoogle Scholar
  5. Cagnolo L, Cabido M, Valladares G (2006) Plant species richness in the Chaco Serrano Woodland from central Argentina: ecological traits and habitat fragmentation effects. Biol Conserv 132:510–519CrossRefGoogle Scholar
  6. Chutima R, Dell B, Vessabutr S, Bussaban B, Lumyong S (2011) Endophytic fungi from Pecteilis susannae (L.) Rafin (Orchidaceae), a threatened terrestrial orchid in Thailand. Mycorrhiza 21:221–229PubMedCrossRefGoogle Scholar
  7. Currah RS, Sigler L, Hambleton S (1987) New records and new taxa of fungi from the mycorrhizae of terrestrial orchids of Alberta. Can J Bot 65:2473–2482CrossRefGoogle Scholar
  8. Dearnaley JDW (2007) Further advances in orchid mycorrhizal research. Mycorrhiza 17:475–486PubMedCrossRefGoogle Scholar
  9. Dressler RL (1993) Phylogeny and classification of the orchid family. Dioscorides, PortlandGoogle Scholar
  10. Dutra D, Kane ME, Richardson L (2009) Asymbiotic seed germination and in vitro seedling development of Cyrtopodium punctatum: a propagation protocol for an endangered Florida native orchid. Plant Cell Tiss Org 96:235–243CrossRefGoogle Scholar
  11. Flachsland EA, Terada G, Rey Y, Mroginski LA (1996) Medios de cultivo para la germinación in vitro de 41 especies de orquídeas. Facena 12:93–100Google Scholar
  12. Flachsland EA, Terada G, Scocchi A, Rey Y, Mroginski LA, Engelmann F (2006) Cryopreservation of seeds and in vitro-cultured protocorms of Oncidium bifolium Sims. (Orchidaceae) by encapsulation-dehydration. Cryo-Lett 27:235–242Google Scholar
  13. Fracchia S, Aranda-Rickert A, Gopar A, Silvani V, Fernandez L, Godeas A (2008) Mycorrhizal status of plant species in the Chaco Serrano Woodland from central Argentina. Mycorrhiza 19:205–214CrossRefGoogle Scholar
  14. Goloboff PA, Farris JS, Nixon KC (2008) TNT, a free program for phylogenetic analysis. Cladistics 24:774–786CrossRefGoogle Scholar
  15. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acid S 41:95–98Google Scholar
  16. Katoh K, Toh H (2008) Recent developments in the MAFFT multiple sequence alignment program. Brief Bioinform 9:286–298PubMedCrossRefGoogle Scholar
  17. Marco DE, Páez SA (2000) Invasion of Gleditsia triacanthos in Lithraea ternifolia Montane Forests of Central Argentina. Environ Manage 26:409–419PubMedCrossRefGoogle Scholar
  18. McCormick M, Whigham D, Sloan D, O'Malley K (2006) Orchid–fungus fidelity: a marriage meant to last? Ecology 87:903–911PubMedCrossRefGoogle Scholar
  19. Newsham KK (2011) A meta-analysis of plant responses to dark septate root endophytes. New Phytol 190:783–793PubMedCrossRefGoogle Scholar
  20. Otero Ospina JT, Bayman P (2009) Germinación simbiótica y asimbiótica en semillas de orquídeas epífitas. Acta Agron 58:270–276Google Scholar
  21. Pereira OL, Megumi Kasuya MC, Rollemberg C, Borges A (2005) In vitro symbiotic seed germination of Oncidium flexosum (Orchidaceae) by Rhizoctonia-like mycorrhizal fungi. R Bras Ci Solo 29:199–206CrossRefGoogle Scholar
  22. Phillips JM, Hayman DS (1970) Improved procedures for clearing roots and staining parasitic and vesicular–arbuscular mycorrhizal fungi for rapid assessment of infection. Trans Brit Mycol Soc 55:158–161CrossRefGoogle Scholar
  23. Porras-Alfaro A, Bayman P (2007) Mycorrhizal fungi of Vanilla: diversity, specificity and effects on seed germination and plant growth. Mycologia 99:510–525PubMedCrossRefGoogle Scholar
  24. Rasmussen HN (1995) Terrestrial orchid: from seed to mycotrophic plant. Cambridge University Press, CambridgeCrossRefGoogle Scholar
  25. Rasmussen HN, Rasmussen FN (2007) Trophic relationships in orchid mycorrhiza—diversity and implications for conservation. Lankesteriana 7:334–341Google Scholar
  26. Schinini A, Waechter J, Izaguirre P, Lehnebach C (2008) Orchidaceae. In: Zuloaga FO, Morrone O, y Belgrano MJ, Catálogo de las Plantas Vasculares del Cono Sur (eds) Monocotyledoneae, vol. 1. St. Louis: Missouri Botanical Garden. pp 472–609Google Scholar
  27. Selosse MA, Faccio A, Scappaticci G, Bonfante P (2004) Chlorophyllous and achlorophyllous specimens of Epipactis microphylla (Neottieae, Orchidaceae) are associated with ectomycorrhizal septomycetes, including truffles. Microb Ecol 47:416–426PubMedCrossRefGoogle Scholar
  28. Sérsic A, Cocucci A, Benítez-Vieyra S, Cosacov A Díaz L, Glinos E, Grosso N, Lazarte C, Medina M, Moré M, Moyano M, Nattero J, Paiaro V, Trujillo C, Wiemer P (2006) Flores del centro de Argentina. 354 pp. Academia Nacional de Ciencias, Córdoba-Argentina. ISBN 987-98313-5-7. CórdobaGoogle Scholar
  29. Sobral A, Fracchia S (2010) Aa achalensis Schltr (Orchidaceae) en la Sierra de Velasco, La Rioja, Argentina. Kurtziana 35:19–21Google Scholar
  30. Steinfort U, Verdugo G, Besoain X, Cisternas MA (2010) Mycorrhizal association and symbiotic germination of the terrestrial orchid Bipinnula fimbriata (Poepp.) Johnst (Orchidaceae). Flora 205:811–817CrossRefGoogle Scholar
  31. Stewart SL, Kane ME (2006) Symbiotic seed germination and in vitro seedling development of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tiss Org 86:147–158CrossRefGoogle Scholar
  32. Stewart S, Kane M (2007) Symbiotic seed germination and evidence for in vitro mycobiont specificity in Spiranthes brevilabris (Orchidaceae) and its implications for species-level conservation. In Vitro Cell Dev B 43:178–186CrossRefGoogle Scholar
  33. Stretton HM, McKenzie AR, Baker KF, Flentje NT (1964) Formation of the basidial stage of some isolates of Rhizoctonia. Phytopathology 54:1093–1095Google Scholar
  34. Urcelay C, Pasquini R, Cánovas S, Liébana V (2005) Colonización micorricica en tres especies de orquídeas nativas de las Sierras de Córdoba, Argentina. Kurtziana 31:51–57Google Scholar
  35. Van Waes JM, Bebergh PC (1986) Adaptation of the tetrazolium method for testing the seed viability, and scanning electron microscopy study of some Western European orchids. Physiol Plant 66:435–442CrossRefGoogle Scholar
  36. Vischi N, Natale E, Villamil C (2004) Six endemic plant species from central Argentina: an evaluation of their conservation status. Biodivers Conserv 13:997–1008CrossRefGoogle Scholar
  37. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic, New York, pp 315–322Google Scholar
  38. Yamazaki J, Miyoshi K (2006) In vitro asymbiotic germination of immature seed and formation of protocorm by Cephalanthera falcata (Orchidaceae). Ann Bot 98:1197–1206PubMedCrossRefGoogle Scholar
  39. Yuan Z, Chen Y, Yang Y (2009) Diverse non-mycorrhizal fungal endophytes inhabiting an epiphytic, medicinal orchid (Dendrobium nobile): estimation and characterization. World J Microb Biot 25:295–303CrossRefGoogle Scholar
  40. Zettler LW, Hofer CJ (1998) Propagation of the little club-spur orchid (Platanthera clavellata) by symbiotic seed germination and its ecological implications. Environ Exp Bot 39:189–195CrossRefGoogle Scholar
  41. Zimmerman E, Peterson RL (2007) Effect of a dark septate fungal endophyte on seed germination and protocorm development in a terrestrial orchid. Symbiosis 43:45–52Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Fracchia Sebastián
    • 1
  • Silvani Vanesa
    • 2
  • Flachsland Eduardo
    • 3
  • Terada Graciela
    • 3
  • Sede Silvana
    • 4
  1. 1.Centro Regional de Investigaciones Científicas y Transferencia Tecnológica, CRILAR- CONICETAnillacoArgentina
  2. 2.Departamento de Biodiversidad y Biología Experimental. Facultad de Ciencias Exactas y Naturales (FCEyN)Universidad de Buenos Aires (UBA)Buenos AiresArgentina
  3. 3.Instituto de Botánica del Nordeste (CONICET—Consejo Nacional de Investigaciones Científicas y Técnicas)Facultad de Ciencias Agrarias (UNNE—Universidad Nacional del Nordeste)CorrientesArgentina
  4. 4.Instituto de Botánica DarwinionBuenos AiresArgentina

Personalised recommendations