Abstract
Vanilla is an orchid of economic importance widely cultivated in tropical regions and native to Mexico. We sampled three species of Vanilla (V. planifolia, V. pompona, and V. insignis) in different crop systems. We studied the effect of crop system on the abundance, type of fungi, and quality of pelotons found in the roots using light and electron microscopy and direct sequencing of mycorrhizal structures. Fungi were identified directly from pelotons obtained from terrestrial roots of vanilla plants in the flowering stage. Root samples were collected from plants in crop systems located in the Totonacapan area in Mexico (states of Puebla and Veracruz). DNA was extracted directly from 40 pelotons and amplified using ITS rRNA sequencing. Peloton-like structures were observed, presenting a combination of active pelotons characterized by abundant hyphal coils and pelotons in various stages of degradation. The most active pelotons were observed in crop systems throughout living tutors (host tree) in comparison with roots collected from dead or artificial tutors. Fungi identified directly from pelotons included Scleroderma areolatum, a common ectomycorrhizal fungus that has not been reported as a mycorrhizal symbiont in orchids. Direct amplification of pelotons also yielded common plant pathogens, including Fusarium and Pyrenophora seminiperda, especially in those sites with low colonization rates, and where large numbers of degraded pelotons were observed. This research reports for the first time the potential colonization of Vanilla by Scleroderma, as a putative orchid mycorrhizal symbiont in four sites in Mexico and the influence of crop system on mycorrhizal colonization on this orchid.
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References
Adame-García J, Rodríguez-Guerra R, Iglesias-Andreu LG, Ramos-Prado JM, Luna-Rodríguez M (2015) Molecular identification and pathogenic variation of Fusarium species isolated from Vanilla planifolia in Papantla Mexico. Bot Sci 93:669–678. 10.17129/botsci.142
Altschul SF, Madden T, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25(17):3389–3402. https://doi.org/10.1093/nar/25.17.3389
Barmicheva KM (1989) Ultrastucture of Neottia nidus-avis mycorrhizas. Agric Ecosyst Environ 29:23–27. https://doi.org/10.1016/0167-8809(90)90248-C
Batty AL, Dixon KW, Brundrett MC, Sivasithamparam K (2002) Orchid conservation and mycorrhizal associations. In: Sivasithamparam K, Dixon KW, Barrett RL (eds) Microorganisms in plant conservation and biodiversity. Springer, Science+Business Media, Dordrecht, pp 195–226
Bayman P, Mosquera-Espinosa AT, Porras-Alfaro A (2011) Mycorrhizal relationship of Vanilla and prospects for biocontrol of root rots. In: Havking-Frenkel D, Belanger FC (eds) Handbook of Vanilla science and technology, 1st edn. Blackwell Publishing Ldt, Oxford, pp 266–280
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. P RoySoc B-Biol Sci 271(1550):1799–1806. https://doi.org/10.1098/rspb.2004.2807
Boddington CL, Dodd JC (1998) A comparison of the development and metabolic activity of mycorrhizas formed by arbuscular mycorrhizal fungi from different genera on two tropical forage legumes. Mycorrhiza 8(3):149–157. https://doi.org/10.1007/s005720050228
Bozzola JJ, Russell LD (1999) Electron microscopy: principles and techniques for biologists, 2nd edn. Jones and Bartlett Publishers, Massachusetts
Chen YL, Kang LH, Malajczuk N, Dell B (2006) Selecting ectomycorrhizal fungi for inoculating plantations in south China: effect of Scleroderma on colonization and growth of exotic Eucalyptus globulus, E. urophylla, Pinus elliottii, and P. radiata. Mycorrhiza 16(4):251–259. https://doi.org/10.1007/s00572-006-0039-8
Cortés-PérezA (2011) Diversidad del género Scleroderma Pers. emend. Fr. (Fungi, Basidiomycotina, Sclerodermatales) en Veracruz. Dissertation, Universidad Veracruzana, Zalapa, Mexico, p 78
Dell B, Malajczuk N, Bougher NL, Thomson G (1994) Development and function of Pisolithus and Scleroderma ectomycorrhizas formed in vivo with Allocasuarina,Casuarina and Eucalyptus. Mycorrhiza 5(2):129−138. https://doi.org/10.1007/s005720050050
Ehlers D, Pfister M (1997) Compounds of vanilla (Vanilla pompona Schiede). J Essent Oil Res 9(4):427–431. https://doi.org/10.1080/10412905.1997.9700743
Filipski A, Kamura S, Peterson D, Stecher G, Tamura K (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197
Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes application to the identification of mycorrhizae and rusts. Mol Ecol 2(2):113–118. https://doi.org/10.1111/j.1365-294X.1993.tb00005.x
Gretzinger B, Dean MD (2011) Vanilla production in the context of culture, economics, and ecology of Belize. In: Havking-Frenkel D, Belanger FC (eds) Handbook of vanilla science and technology, 1st edn. Blackwell Publishing Ltd, Oxford, pp 50–68
Guzmán G, Cortés-Pérez A, Guzmán-Dávalos L, Ramírez-Guillén F, Sánchez-Jácome MR (2013) An emendation of Scleroderma, new records, and review of the known species in Mexico. Rev Mex Biodivers 84:S191–S191. https://doi.org/10.7550/rmb.31979
Havkin-Frenkel D, Dorn R (1997) Vanilla. In: Risch J, Ho CT (eds) Species, flavor chemistry and antioxidant properties. American Chemical Society, Washington DC, pp 29–40. https://doi.org/10.1021/bk-1997-0660.ch004
Havkin-Frenkel D, Belanger FC (2011) Handbook of vanilla science and technology,1st edn. Blackwell Publishing Ltd, Oxford
Huynh TT, Lawrie AC, Coates F, McLean CB (2004) Effect of developmental stage and peloton morphology on success in isolation of mycorrhizal fungi in Caladenia formosa (Orchidaceae). Aust J Bot 52:231–241. https://doi.org/10.1071/bt03099
IMPI (Instituto Mexicano de la Propiedad Industrial) (2016) Denominaciones de origen. In: Orgullo de México. Secretaría de Economía, México
Kohout P, Tĕšitelova T, Roy M, Vohnik M, Jersáková J (2013) A diverse fungal community associated with Pseudorchis albida (Orchidaceae) roots. Fungal Ecol 6(1):50–64. https://doi.org/10.1016/j.funeco.2012.08.005
Kõljalg U, Nilsson RH, Abarenkov K, Tedersoo L, Taylor AFS, Bahram M, Bates ST, Bruns TD, Bengtsson-Palme J, Callaghan TM, Douglas B, Drenkhan T, Eberhardt U, Dueñas M, Grebenc T, Griffith GW, Hartmann M, Kirk PM, Kohout P, Larsson E, Lindahl BD, Lücking R, Martín MP, Matheny PB, Nguyen NH, Niskanen T, Oja J, Peay KG, Peintner U, Peterson M, Põldmaa K, Saag L, Saar I, Schüßler A, Scott JA, Senés C, Smith ME, Suija A, Taylor DL, Telleria MT, Weiß M, Larsson KH (2013) Towards a unified paradigm for sequence-based identification of fungi. Mol Ecol 22(21):5271–5277. https://doi.org/10.1111/mec.12481
Kristiansen KA, Taylor DL, Kjøller R, Rasmussen HN, Rosendahl S (2001) Identification of mycorrhizal fungi from single pelotons of Dactylorhiza majalis (Orchidaceae) using single-strand conformation polymorphism and mitochondrial ribosomal large subunit DNA sequences. Mol Ecol 10(8):2089–2093. https://doi.org/10.1046/j.0962-1083.2001.01324.x
Kuga Y, Sakamoto N, Yurimoto H (2014) Stable isotope cellular imaging reveals that both live and degenerating fungal pelotons transfer carbon and nitrogen to orchid protocorms. New Phytol 202(2):594–605. https://doi.org/10.1111/nph.12700
Làtr A, Čuříková M, Baláž M, Jurčák J (2008) Mycorrhizas of Cephalanthera longifolia and Dactylorhiza majalis, two terrestrial orchids. Ann Bot Fenn 45(4):281–289. https://doi.org/10.5735/085.045.0405
Leake JR, Cameron DD (2012) Untangling aboveand belowground mycorrhizal fungal networks in tropical orchids. Mol Ecol 21(20):921–4924. https://doi.org/10.1111/j.1365-294X.2012.05718.x
Lee S-S (2002) A review of orchid mycorrhizae in Korea. Plant Pathol J 18(4):169–178. https://doi.org/10.5423/PPJ.2002.18.4.169
Lee YI, Yang CK, Gebauer G (2015) The importance of associations with saprotrophic non-Rhizoctonia fungi among fully mycoheterotrophic orchids is currently under-estimated: novel evidence from sub-tropical Asia. Ann Bot 116(3):423–435. https://doi.org/10.1093/aob/mcv085
Lubinsky P, Romero-Gonzalez GA, Heredfia SM, Zabel S (2011) Origins and patterns of vanilla cultivation in tropical America (1500−1900): no support for an independent domestication of vanilla in South America. In: Havking-Frenkel D, Belanger FC (eds) Handbook of Vanilla science and technology, 1st edn. Blackwell Publishing Ldt, Oxford, pp 117–138
Masuhara G, Katsuya K (1994) In situ and in vitro specificity between Rhizoctonia spp. and Spiranthessinensis (Persoon) Ames, var. amoena (M. Bieberstein) hara (Orchidaceae). New Phytol 127(4):711–718. https://doi.org/10.1111/j.1469-8137.1994.tb02974.x
Mejstrik V (1970) The anatomy of roots and mycorrhizae of the orchid Dendrobium cunninhamii Lindl. Biol Plantarum 12(2):105–109. https://doi.org/10.1007/BF02920857
Moncalvo JM, Nilsson RH, Koster B, Dunham SM, Bernauer T, Matheny PB, Porter TM, Margaritescu S, Weiß GS, Danell E, Langer G, Langer E, Larsson E, Larsson KH, Vilgalys R (2006) The cantharelloid clade: dealing with incongruent gene trees and phylogenetic reconstruction methods. Mycologia 98(6):937–948. https://doi.org/10.1080/15572536.2006.11832623
Nouhra E, Hernández M, Pastor N, Crespo E (2012) The species of Scleroderma from Argentina, including a new species from the Nothofagus forest. Mycologia 104(2):488–495. https://doi.org/10.3852/11-082
Oja J, Kohout P, Tedersoo L, Kull T, Kõljalg U (2015) Temporal patterns of orchid mycorrhizal fungi in meadows and forests as revealed by 454 pyrosequencing. New Phytol 205(4):1608–1618. https://doi.org/10.1111/nph.13223
Otero JT, Bayman P (2009) Germinación simbiótica y asimbiótica en semillas de orquídeas epífitas. Acta Agronómica 58:270–276. 10.15446/acag
Otero JT, Ackerman JD, Bayman P (2002) Diversity and host specificity of endophytic Rhizoctonia-like fungi from tropical orchids. Am J Bot 89(11):1852–1858. https://doi.org/10.3732/ajb.89.11.1852
Pathan AK, Bond J, Gaskin RE (2010) Sample preparation for SEM of plant surfaces. Mater Today 12:32–43. https://doi.org/10.1016/S1369-7021(10)70143-7
Pellegrino G, Bellusci F (2009) Molecular identification of mycorrhizal fungi in Dactylorhiza sambucina (Orchidaceae). Biologia 64:893–897. https://doi.org/10.2478/s11756-009-0175-7
Pereira OL, Kasuya MCM, Borges AC, Araújo EF (2005) Morphological and molecular characterization of mycorrhizal fungi isolated from neotropical orchids in Brazil. Can J Botany 83(1):54–65. https://doi.org/10.1139/b04-151
Peterson RL, Massicotte HB (2004) Exploring structural definitions of mycorrhizas, with emphasis on nutrient-exchange interfaces. Can J Botany 82(8):1074–1088. https://doi.org/10.1139/b04-071
Porras-Alfaro A, Bayman P (2003) Mycorrhizal fungi of Vanilla: root colonization patterns and fungal identification. Lankesteriana 7:147–150. 10.15517/lank.v3i2.23041
Porras-Alfaro A, Bayman P (2007) Mycorrhizal fungi of Vanilla: diversity, specificity and effects on seed germination and plant growth. Mycologia 99(4):510–525. https://doi.org/10.1080/15572536.2007.11832545
Rasmussen HN (1995) Terrestrial orchids: from seed to mycotrophic plant. Cambridge University Press, New York. https://doi.org/10.1017/CBO9780511525452
Rasmussen HN (2002) Recent developments in the study of orchid mycorrhiza. Plant Soil 244(1/2):149–163. https://doi.org/10.1023/A:1020246715436
Rasmussen HN, Rasmussen FN (2014) Seedling mycorrhiza: a discussion of origin and evolution in Orchidaceae. Bot J Linn Soc 175(3):313–327. https://doi.org/10.1111/boj.12170
Rasmussen HN, Whigham DF (2002) Phenology of roots and mycorrhizae in orchid species differing in phototrophic strategy. New Phytol 154(3):797–807. https://doi.org/10.1046/j.1469-8137.2002.00422.x
Roche SA, Carter RJ, Peakall R, Smith LM, Whitehead MR, Linde CC (2010) A narrow group of monophyletic Tulasnella (Tulasnellaceae) symbiont lineages are associated with multiple species of Chiloglottis (Orchidaceae): implications for orchid diversity. Am J Bot 97(8):1313–1327. https://doi.org/10.3732/ajb.1000049
Roy M, Yagame T, Yamato M, Iwase K, Heinz C, Faccio A, Bonfante P, Selosse MA (2009) Ectomycorrhizal Inocybe species associate with the mycoheterotrophic orchid Epipogium aphyllum but not its asexual propagules. Ann Bot-London 104(3):595–610. https://doi.org/10.1093/aob/mcn269
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. https://doi.org/10.1007/s00248-003-2034-3
Shefferson RP, Taylor DL, Weiss M, Garnica S, McCormick MK, Adams S, Gray HM, McFarland JW, Kull T, Tali K, Yukawa T, Kawahara T, Miyoshi K, Lee YI (2007) The evolutionary history of mycorrhizal specificity among lady's slipper orchids. Evolution 61(6):1380–1390. https://doi.org/10.1111/j.1558-5646.2007.00112.x
Shefferson RP, Kull T, Tali K (2008) Mycorrhizal interactions of orchids colonizing Estonian mine tailings hills. Am J Bot 95(2):156–164. https://doi.org/10.3732/ajb.95.2.156
Smith SE, Read DJ (1997) Mycorrhizal symbiosis, 2nd edn. Academic Press, California
Sreedhar RV, Venkatachalam L, Roohie K, Bhagyalaksmi N (2007) Molecular analysis of Vanilla planifolia cultivated in India using RAPD and ISSR markers. Orchid Sci Biotechnol 1:29–33
Taylor DL, Bruns TD, Leake JR, Read DJ (2002) Mycorrhizal specificity and function in myco-heterotrophic plants. In: van der Heijden MGA, Sanders RI (eds) The ecology of mycorrhizas. Springer Verlag, Berlin, pp 375–414. https://doi.org/10.1007/978-3-540-38364-2_15
Taylor DL, McCormick MK (2008) Internal transcribed spacer primers and sequences for improved characterization of basidiomycetous orchid mycorrhizas. New Phytol 177(4):1020–1033. https://doi.org/10.1111/j.1469-8137.2007.02320.x
Valadares RB, Pereira MC, Otero JT, Cardoso EJ (2012) Narrow fungal mycorrhizal diversity in a population of the orchid Coppensia doniana. Biotropica 44(1):114–122. https://doi.org/10.1111/j.1744-7429.2011.00769.x
White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR protocols: a guide to methods and applications. Academic press Inc, New York, pp 315–322
Yamada A, Katsuya K (1995) Mycorrhizal association of isolates from sporocarps and ectomycorrhizas with Pinus densiflora seedlings. Mycoscience 36(3):315–323. https://doi.org/10.1007/BF02268607
Zettler LW (1997) Terrestrial orchid conservation by symbiotic seed germination: techniques and perspectives. Selbyana 18:188–194.
Zettler LW, Stewart SL, Bowles ML, Jacobs KA (2001) Mycorrhizal fungi and cold-assisted symbiotic germination of the federally threatened eastern prairie fringed orchid, Platanthera leucophaea (Nuttall) Lindley. Am Midl Nat 145(1):168–175. https://doi.org/10.1674/0003-0031(2001)145[0168:MFACAS]2.0.CO;2
Zhu GS, ZN Y, Gui Y, Liu ZY (2008) A novel technique for isolating orchid mycorrhizal fungi. Fungal Divers 33:123–137
Acknowledgements
Authors thank SAGARPA-CONACYT-SNITT fund for their support through the Subproject 03 and Megaproject 2012-04-190442; and Vanilla producers for the opportunity to sample in their commercial plots. SAS thanks for her scholarship from this project.
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González-Chávez, M.d.C.A., Torres-Cruz, T.J., Sánchez, S.A. et al. Microscopic characterization of orchid mycorrhizal fungi: Scleroderma as a putative novel orchid mycorrhizal fungus of Vanilla in different crop systems. Mycorrhiza 28, 147–157 (2018). https://doi.org/10.1007/s00572-017-0808-6
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DOI: https://doi.org/10.1007/s00572-017-0808-6