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Preliminary findings on identification of mycorrhizal fungi from diverse orchids in the Central Highlands of Madagascar

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Abstract

The Orchid flora of Madagascar is one of the most diverse with nearly 1000 orchid taxa, of which about 90 % are endemic to this biodiversity hotspot. The Itremo Massif in the Central Highlands of Madagascar with a Highland Subtropical climate range encompasses montane grassland, igneous and metamorphic rock outcrops, and gallery and tapia forests. Our study focused on identifying culturable mycorrhizae from epiphytic, lithophytic, and terrestrial orchid taxa to understand their diversity and density in a spatial matrix that is within the protected areas. We have collected both juvenile and mature roots from 41 orchid taxa for isolating their orchid mycorrhizal fungi (OMF), and to culture, identify, and store in liquid nitrogen for future studies. Twelve operational taxonomic units (OTUs), of three known orchid mycorrhizal genera, were recognized by analysis of internal transcribed spacer (ITS) sequences of 85 isolates, and, by comparing with GenBank database entries, each OTU was shown to have closely related fungi that were also found as orchid associates. Orchid and fungal diversity were greater in gallery forests and open grasslands, which is very significant for future studies and orchid conservation. As far as we know, this is the first ever report of detailed identification of mycorrhizal fungi from Madagascar. This study will help start to develop a programme for identifying fungal symbionts from this unique biodiversity hotspot, which is undergoing rapid ecosystem damage and species loss. The diversity of culturable fungal associates, their density, and distribution within the Itremo orchid hotspot areas will be discussed.

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References

  • Alvarado ST, Buisson E, Rabarison H, Rajeriarison C, Birkinshaw C, Lowry PP (2014) Comparison of plant communities on the Ibity and Itremo massifs, Madagascar, with contrasting conservation histories and current status. Plant Ecol Divers 7:497–508

  • Arditti J (1992) Fundamentals of orchid biology. Wiley, New York

    Google 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. Aust J Bot 54:375–381

    Article  Google Scholar 

  • Bidartondo MI, 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 B 271:1799–1806

    Article  CAS  Google Scholar 

  • Bonnardeaux Y, Brundrett M, Batty A, Dixon K, Koch J, Sivasithamparam K (2007) Diversity of mycorrhizal fungi of terrestrial orchids: compatibility webs, brief encounters, lasting relationships and alien invasions. Mycol Res 111:51–61

    Article  PubMed  Google Scholar 

  • Clements MA, Ellyard RK (1979) The symbiotic germination of Australian terrestrial orchids. Am Orchid Soc Bull 48:810–816

    Google Scholar 

  • Cribb P, Hermans J (2009) Field guide to the orchids of Madagascar. Kew Publishing. Royal Botanic Gardens, Kew

    Google Scholar 

  • Dearnaley JDW (2007) Further advances in orchid mycorrhizal research. Mycorrhiza 17:475–486

    Article  PubMed  Google Scholar 

  • Dixon KW (1987) Raising terrestrial orchids from seed. In: Harris WK (ed) Modern orchid growing for pleasure and profit. Orchid Club of S Australia, Inc, Adelaide, pp 47–100

    Google Scholar 

  • Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15

    Google Scholar 

  • du Puy D, Moat J (1996) A refined classification of the primary vegetation of Madagascar based on the underlying geology: using GIS to map its distribution and to assess its conservation status. In: Lourenço WR (ed) Proceedings of the International Symposium on the ‘Biogeography de Madagascar’, Paris, September 1995, pp 205–218

  • Fenu G, Mattana E, Congiu A, Bacchetta G (2010) The endemic vascular flora of Supramontes (Sardinia), a priority plant conservation area. Candollea 65:347–358

    Article  Google Scholar 

  • Gale SW, Yamazaki J, Hutchings MJ, Yukawa T, Miyoshi K (2010) Constraints on establishment in an endangered terrestrial orchid: a comparative study of in vitro and in situ seed germinability and seedling development in Nervilia nipponica. Bot J Lin Soc 163:166–180

    Article  Google Scholar 

  • Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes: application to the identification of mycorrhizae and rusts. Mol Ecol 2:113–118

    Article  CAS  PubMed  Google Scholar 

  • Girlanda M, Segreto R, Cafasso D, Liebel HT, Rodda M, Ercole E, Cozzolino S, Gebauer G, Perotto S (2011) Photosynthetic Mediterranean meadow orchids feature partial mycoheterotrophy and specific mycorrhizal associations. Am J Bot 98:1148–1163

    Article  PubMed  Google Scholar 

  • Jacquemyn H, Deja A, De hert K, Bailarote BC, Lievens B (2012) Variation in mycorrhizal associations with tulasnelloid fungi among populations of five Dactylorhiza species. PLoS One 7(8):e42212

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Jonsson L, Nylund JE (1979) Favolaschia dybowskyana (Singer) Singer (Aphyllophorales), a new orchid mycorrhizal fungus from tropical Africa. New Phytol 83:121–128

    Article  Google Scholar 

  • Martos F, Munoz F, Pailler T, Kottke I, Gonneau C, Selosse MA (2012) The role of epiphytism in architecture and evolutionary constraint within mycorrhizal networks of tropical orchids. Mol Ecol 21:5098–5109

    Article  PubMed  Google Scholar 

  • McKendrick SL, Leake JR, Taylor DL, Read DJ (2000) Symbiotic germination and development of myco-heterotrophic plants in nature: ontogeny of Corallorhiza trifida and characterization of its mycorrhizal fungi. New Phytol 145:523–537

    Article  Google Scholar 

  • Mitchell RB (1989) Growing hardy orchids from seeds at Kew. Plantsman 2:152–169

    Google Scholar 

  • Mittermeier RA, Robles PG, Hoffman M, Pilgrim J, Brooks T, Mittermeier CG, Lamoreux J, da Fonseca GAB (2005) Hotspots revisited: earth’s biologically richest and most endangered terrestrial ecoregions. University of Chicago Press, Chicago

    Google Scholar 

  • Moat J, Smith P (2007) Atlas of the vegetation of Madagascar. Royal Botanic Gardens, Kew

    Google Scholar 

  • Mugambi GK (2001) Ensuring survival of Kenyan orchids: the ex situ conservation interventions. International Orchid Conservation Congress I (Conference Proceedings), p 102

  • Murray-Smith C, Brummitt NA, Oliviera-Filho AT, Bachman S, Moat J, Lughadha EM, Lucas EJ (2009) Plant diversity hotspots in the Atlantic coastal forests of Brazil. Conserv Biol 23:151–163

    Article  PubMed  Google Scholar 

  • Nilsson LA, Rabakonandrianina E (1988) Hawk-moth scale analysis and pollination specialization in the epilithic malagasy endemic Aerangis ellisii (Reichenb fil) Schltr (Orchidaceae). Bot J Linn Soc 97:49–61

    Article  Google Scholar 

  • Nilsson LA, Rabakonandrianina E, Pettersson B (1992a) Exact tracking of pollen transfer and mating in plants. Nature 360:666–668

    Article  Google Scholar 

  • Nilsson LA, Rabakonandrianina E, Rotaharivelo R, Randriamanindry JJ (1992b) Long pollinia on eyes: hawk-moth pollination of Cynorkis uniflora Lindley (Orchidaceae) in Madagascar. Bot J Linn Soc 109:145–160

    Article  Google Scholar 

  • Nontachaiyapoom S, Sasirat S, Manoch L (2010) Isolation and identification of Rhizoctonia-like fungi from roots of three orchid genera, Paphiopedilum, Dendrobium, and Cymbidium, collected in Chiang Rai and Chiang Mai provinces of Thailand. Mycorrhiza 20:459–471

    Article  PubMed  Google Scholar 

  • Otero JT, Ackerman JD, Bayman P (2002) Diversity and host specificity of endophytic Rhizoctonia-like fungi from tropical orchids. Am J Bot 89:1852–1858

    Article  CAS  Google Scholar 

  • Phillips RD, Barrett MD, Dixon KW, Hopper SD (2011) Do mycorrhizal symbioses cause rarity in orchids? J Ecol 99:858–869

    Article  Google Scholar 

  • Rasmussen HN (2002) Recent developments in the study of orchid mycorrhiza. Plant Soil 244:149–163

    Article  CAS  Google Scholar 

  • Rasmussen HN, Whigham DF (1993) Seed ecology of dust seeds in situ: a new study technique and its application to terrestrial orchids. Am J Bot 80:1374–1378

    Article  Google Scholar 

  • Richardson KA, Currah RS, Hambleton S (1993) Basidiomycetous endophytes from the roots of neotropical epiphytic Orchidaceae. Lindleyana 8:127–137

    Google Scholar 

  • Roos MC, Kessler PJA, Gradstein SR, Baas P (2004) Species diversity and endemism of five major Malesian islands: diversity–area relationships. J Biogeogr 31:1893–1908

    Article  Google Scholar 

  • Samarakoon T, Wang SY, Alford MH (2013) Enhancing PCR amplification of DNA from recalcitrant plant specimens using a trehalose-based additive. Appl Plant Sci 1:1200236

    Google Scholar 

  • 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 47:416–426

    CAS  PubMed  Google Scholar 

  • Swarts ND, Dixon KW (2009) Terrestrial orchid conservation in the age of extinction. Ann Bot 104:543–556

    Article  PubMed Central  PubMed  Google Scholar 

  • Taberlet P, Gielly L, Pautou G, Bouvet J (1991) Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol Biol 17:1105–1109

    Article  CAS  PubMed  Google Scholar 

  • Taylor DL, McCormick MK (2008) Internal transcribed spacer primers and sequences for improved characterization of basidiomycetous orchid mycorrhizas. New Phytol 177:1020–1033

    Article  CAS  PubMed  Google Scholar 

  • Tyson P (2000) The eighth continent: life, death and discovery in the lost world of Madagascar. William Morrow (Harper Collins) Publishers, New York

    Google Scholar 

  • Vorontsova MS, Ratovonirina G, Randriamboavonjy T (2013) Revision of Andropogon and Diectomis (Poaceae: Sacchareae) in Madagascar and the new Andropogon itremoensis from the Itremo Massif. Kew Bull 68:1–15

    Article  Google Scholar 

  • Warcup JH, Talbot PHB (1980) Perfect states of Rhizoctonias associated with orchids III. New Phytol 86:267–272

    Article  Google Scholar 

  • Waterman RJ, Bidartondo MI, Stofberg J, Combs JK, Gebauer G, Savolainen V, Barraclough TG, Pauw A (2011) The effects of above- and belowground mutualisms on orchid speciation and coexistence. Am Nat 177:54–68

    Article  Google Scholar 

  • Weiss M, Selosse MA, Rexer KH, Urban A, Oberwinkler F (2004) Sebacinales: a hitherto overlooked cosm of heterobasidiomycetes with a broad mycorrhizal potential. Mycol Res 108:1003–1010

    Article  PubMed  Google Scholar 

  • White TJ, Bruns TD, Lee S, Taylor JW (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, San Diego, pp 315–322

    Google Scholar 

  • Whitman M, Medler M, Randriamanindry JJ, Rabakonandrianina E (2011) Conservation of Madagascar’s granite outcrop orchids: the influence of fire and moisture. Lankesteriana 11:55–67

    Article  Google Scholar 

  • Zettler LW, Piskin KA (2011) Mycorrhizal fungi from protocorms, seedlings and mature plants of the Eastern Prairie Fringed Orchid, Platanthera leucophaea (Nutt.) Lindl.: a comprehensive list to augment conservation. Am Midl Nat 166:29–39

    Article  Google Scholar 

  • Zettler LW, Sharma J, Rasmussen F (2003) Mycorrhizal diversity. In: Dixon KW, Kell SP, Barrett RL, Cribb PJ (eds) Orchid conservation. Natural History Publications (Borneo), Kota, pp 185–203

    Google Scholar 

  • Zettler LW, Corey LL, Jacks AL, Gruender LT, Lopez AM (2013) Tulasnella irregularis (Basidiomycota: Tulasnellaceae) from roots of Encyclia tampensis in south Florida, and confirmation of its mycorrhizal significance through symbiotic seed germination. Lankesteriana 13:119–128

    Google Scholar 

  • Zi XM, Sheng CL, Goodale UM, Shao SC, Gao JY (2014) In situ seed baiting to isolate germination-enhancing fungi for an epiphytic orchid, Dendrobium aphyllum (Orchidaceae). Mycorrhiza 24:487–499

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We kindly thank the financial support received from Sainsbury Orchid Project, Bentham and Moxon Trust, and Margaret A Cargill Foundation. We acknowledge the invaluable assistance received from Gaëtan Ratovonirina and Landy Rajaovelona (KMCC), Solo Rapanarivo (PBZT) for field support during the collection; Korrie Edwards (Illinois College), and Helen Sandford, Margaret Ramsay, and Edward Jones (Kew) for technical assistance; Connie Gibas and Lynne Sigler (UAMH) for deposition of isolates; Stuart Cable (Kew) and Hanne Rasmussen for helpful suggestions; and Mike Fay and Robyn Cowan (Kew) for their advice on genetic fingerprinting of plant samples.

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Authors and Affiliations

  1. Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3AB, UK

    Kazutomo Yokoya, Jonathan P. Kendon, Martin I. Bidartondo & Viswambharan Sarasan

  2. Department of Biology, Illinois College, 1101 West College Avenue, Jacksonville, IL, 62650, USA

    Lawrence W. Zettler, Andrew L. Stice, Shannon Skarha, Laura L. Corey & Audrey C. Knight

  3. Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK

    Martin I. Bidartondo

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  1. Kazutomo Yokoya
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  2. Lawrence W. Zettler
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  4. Martin I. Bidartondo
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  9. Viswambharan Sarasan
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Correspondence to Viswambharan Sarasan.

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Yokoya, K., Zettler, L.W., Kendon, J.P. et al. Preliminary findings on identification of mycorrhizal fungi from diverse orchids in the Central Highlands of Madagascar. Mycorrhiza 25, 611–625 (2015). https://doi.org/10.1007/s00572-015-0635-6

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