Abstract
Species from Ceratobasidium are known as orchid mycorrhizal, plant pathogens or saprophytic. In mycorrhizal association with orchids, Ceratobasidium can establish a highly specific interaction. Gomesa recurva is an epiphytic orchid occurring in the Brazilian Atlantic Forest. In the present study mycorrhizal fungi were isolated from roots of G. recurva and its phylogenetic positions was investigated. A total of nine isolates were obtained directly from pelotons and submitted to phylogenetic analysis based on alignments of sequences of the internal transcribed spacer (ITS) of the nuclear ribosomal DNA. Phylogenetic analyzes, along with morphological data, revealed that isolates are distinct from all known Ceratobasidium species and a new species, namely Ceratobasidium gomesae sp. nov., is proposed. Given the dependence on mycorrhizae for germination and protocorm development, knowledge about orchid symbionts is an important factor in managing the conservation of these plants, and future studies may investigate the ability of C. gomesae to promote seed germination.
References
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Mendes, M.A.S. & Urben, A.F. (2021) Fungos relatados em plantas no Brasil, Laboratório de Quarentena Vegetal. Brasília, DF: Embrapa Recursos Genéticos e Biotecnologia. Available from: http://pragawall.cenargen.embrapa.br/aiqweb/michtml/fgbanco01.asp (accessed 22 August 2021)
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Miller, M.A., Pfeiffer, W. & Schwartz, T. (2010) Creating the CIPRES science gateway for inference of large phylogenetic trees. Gateway Computing Environments Workshop: 1–8. https://doi.org/10.1109/GCE.2010.5676129
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Otero, J.T., Ackerman, J.D. & Bayman, P. (2004) Differences in mycorrhizal preferences between two tropical orchids. Molecular Ecology 13: 2393–2404. https://doi.org/10.1111/j.1365-294X.2004.02223.x
Otero, J.T., Flanagan, N.S., Herre, E.A., Ackerman, J.D. & Bayman, P. (2007) Widespread mycorrhizal specificity correlates to mycorrhizal function in the neotropical, epiphytic orchid Ionopsis utricularioides (Orchidaceae). American Journal of Botany 94 (12): 1944–1950. https://doi.org/10.3732/ajb.94.12.1944
Pereira, O.L., Rollemberg, C.L., Borges, A.C., Kasuya, M.C.M. & Matsuoka, K. (2003) Epulorhiza epiphytica sp. nov. isolated from mycorrhizal roots of epiphytic orchids in Brazil. Mycoscience 44 (2): 153–155. https://doi.org/10.1007/S10267-002-0087-7
Pereira, O.L., Kasuya, M.C.M., Borges, A.C. & Araújo, E.F. (2005) Morphological and molecular characterization of mycorrhizal fungi isolated from neotropical orchids in Brazil. Canadian Journal Botany 83: 54–65. https://doi.org/10.1139/b04-151
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Rasmussen, H.N. (2002) Recent developments in the study of orchid mycorrhiza. Plant and Soil 244 (1–2): 149–163. https://doi.org/10.1023/A:1020246715436
Ronquist, F. & Huelsenbeck, J.P. (2003) MrBayes 3: Beyesian phylogenetic inference under mixed models. Bioinformatics 19: 1572–1574. https://doi.org/10.1093/bioinformatics/btg180
Rambaut, A. (2009) FigTree version 1.4.4. Available from: http://tree.bio.ed.ac.uk/software/figtree/ (accessed 5 February 2020)
Rammitsu, K., Yagame, T., Yamashita, Y., Yukawa, T., Isshiki, S. & Ogura-Tsujita, Y. (2019) A leafless epiphytic orchid, Taeniophyllum glandulosum Blume (Orchidaceae), is specifically associated with the Ceratobasidiaceae family of basidiomycetous fungi. Mycorrhiza 29 (2): 159–166. https://doi.org/10.1007/s00572-019-00881-7
Rannala, B. & Yang, Z. (1996) Proballity distribution of molecular evolutionary trees: A new method of phylogenetic inference. Journal of Molecular Evolution 43: 304–311. https://doi.org/10.1007/BF02338839
Smith, S.E. & Read, D.J. (2008) Mycorrhizal Symbiosis, 3rd edn. Adelaide, Australia.
Thixton, H.L., Esselman, E.J., Corey, L.L. & Zettler, L.W. (2020) Further evidence of Ceratobasidium D.P. Rogers (Basidiomycota) serving as the ubiquitous fungal associate of Platanthera leucophaea (Orchidaceae) in the North American tallgrass prairie. Botanical Studies 61: 12. https://doi.org/10.1186/s40529-020-00289-z
Valadares, R.B., Pereira, M.C., Otero, J.T. & Cardoso, E.J. (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
Warcup, J.H. & Talbot, P.H.B. (1980) Perfect states of Rhizoctonias associated with orchids. New Phytologist 66 (4): 631–641. https://doi.org/10.1111/j.1469-8137.1980.tb00787.x
Weiß, M., Waller, F., Zuccano, A. & Selosse, M.A. (2016) Sebacinales – one thousand and one interactions with land plants. New Phytologist 211: 20–40. https://doi.org/10.1111/nph.13977
White, T.J., Bruns, T.D., Lee, S. & Taylor, J. (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis, M.A., Gelfand, D.H., Sninsky, J.J. & White, T.J. (Eds.) PCR protocols: a guide to methods and applications. Academic, San Diego, pp. 315–322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
Zettler, L.W., Piskin, K.A., Stewart, S.L., Hartsock, J.J., Bowles, M.L. & Bell, T.J. (2005) Protocorm mycobionts of the Federally threatened Eastern Prairie Fringed Orchid, Platanthera leucophaea (Nutt.) Lindl., and a technique to prompt leaf elongation in seedlings. Studies in Mycology 53: 163–171. https://doi.org/10.3114/sim.53.1.163
Ceresini, P.C., Sousa, E.C., Zala, M., Furtado, E.L. & Souza, N.L. (2012) Evidence that the Ceratobasidium-like white-thread blight and black rot fungal pathogens from persimmon and tea crops in the Brazilian Atlantic Forest agroecosystem are two distinct phylospecies. Genetics and Molecular Biology 35 (2): 480–497. https://doi.org/10.1590/S1415-47572012005000032
Costa, R.C., Verzignassi, J.R., Poltronieri, T.P.S., Poltronieri, L.S. & Monteiro, L.C. (2013) Novos hospedeiros de Ceratobasidium ochroleucum, agente causal da queima-do-fio, no Pará. Summa Phytopathologica 39: 62. https://doi.org/10.1590/S0100-54052013000100011
Dearnaley, J.D. (2007) Further advances in orchid mycorrhizal research. Mycorrhiza 17 (6): 475–486. https://doi.org/10.1007/s00572-007-0138-1
Ferreira, B.W., Nóbrega, T.F. & Barreto, R.W. (2021) Reinstating Ceratobasidium lantanae-camarae: the white thread blight fungus on the pantropical weed Lantana camara. Australasian Plant Pathology 50: 545–557. https://doi.org/10.1007/s13313-021-00790-4
Freitas, E.F.S., Silva, M., Cruz, E.S., Mangaravite, E., Bocayuva, M.F., Veloso, T.G.R., Selosse, M.A. & Kasuya, M.C.M. (2020) Diversity of mycorrhizal Tulasnella associated with epiphytic and rupicolous orchids from the Brazilian Atlantic Forest, including four new species. Scientific Reports 10 (1): 1–14. https://doi.org/10.1038/s41598-020-63885-w
Gao, Y., Zhao, Z., Li, J., Liu, N., Jacquemyn, H., Guo, S. & Xing, X. (2020) Do fungal associates of co-occurring orchids promote seed germination of the widespread orchid species Gymnadenia conopsea? Mycorrhiza 30: 221–228. https://doi.org/10.1007/s00572-020-00943-1
Govaerts, R., Bernet, P., Kratochvil, K., Gerlach, G., Carr, G., Alrich, P., Pridgeon, A.M., Pfahl, J., Campacci, M.A., Holland Baptista, D., Tigges, H., Shaw, J., Cribb, P.J., George, A., Kreuz, K. & Wood, J. (2021) World Checklist of Orchidaceae. Facilitated by the Royal Botanic Gardens, Kew. Available from: http://wcsp.science.kew.org/ (accessed September 2021)
Gónzalez, D., Rodriguez-Carres, M., Boekhout, T., Stalpers, J., Kuramae, E.E., Nakatani, A.K., Vilgalys, R. & Cubeta, M.A. (2016) Phylogenetic relationships of Rhizoctonia fungi within the Cantharellales. Fungal Biology 120 (4): 603–619. https://doi.org/10.1016/j.funbio.2016.01.012
Graham, R.R. & Dearnaley, J.D.W. (2012) The rare Australian epiphytic orchid Sarcochilus weinthalii associates with a single species of Ceratobasidium. Fungal Diversity 54 (1): 31–37. https://doi.org/10.1007/s13225-011-0106-0
IUCN (2021) The IUCN Red List of Threatened Species. Version 2021-2. Available from: https://www.iucnredlist.org (accessed 5 November 2021)
Katoh, K. & Standley, D.M. (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution 30 (4): 772–780. https://doi.org/10.1093/molbev/mst010
Kumar, S., Stecher, G. & Tamura, K. (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33: 1870–1874. https://doi.org/10.1093/molbev/msw054
Macedo, D.M., Pereira, O.L., Hora Júnior, B.T., WEIR, B.S. & Barreto, R.W. (2016) Mycobiota of the weed Tradescantia fluminensis in its native range in Brazil with particular reference to classical biological control. Australasian Plant Pathology 45: 45–56. https://doi.org/10.1007/s13313-015-0388-x
Meinhardt, L.W., Bellato, C.D.M. & Tsai, S.M. (2001) SYBR® Green I used to evaluate the nuclei number of fungal mycelia. Biotechniques 31: 42–46. https://doi.org/10.2144/01311bm06
Melo, M.P., Matos, K.S., Moreira, S.I., Silva, F.F., Conceição, G.H., Nechet, K.L., Halfeld-Vieira, B.A., Bezerra Júnior, J.E.A., Ventura, J.A., Costa, H., Furtado, E.L., Alves, E. & Ceresini, P.C. (2018) Two new Ceratobasidium species causing white thread blight on tropical plants in Brazil. Tropical Plant Pathology 43 (6): 559–571. https://doi.org/10.1007/s40858-018-0237-x
Mendes, M.A.S. & Urben, A.F. (2021) Fungos relatados em plantas no Brasil, Laboratório de Quarentena Vegetal. Brasília, DF: Embrapa Recursos Genéticos e Biotecnologia. Available from: http://pragawall.cenargen.embrapa.br/aiqweb/michtml/fgbanco01.asp (accessed 22 August 2021)
Meng, Y.Y., Zhang, W.L., Selosse, M.A. & Gao, J.Y. (2019) Are fungi from adult orchid roots the best symbionts at germination? A case study. Mycorrhiza 29: 541–547. https://doi.org/10.1007/s00572-019-00907-0
Miller, M.A., Pfeiffer, W. & Schwartz, T. (2010) Creating the CIPRES science gateway for inference of large phylogenetic trees. Gateway Computing Environments Workshop: 1–8. https://doi.org/10.1109/GCE.2010.5676129
Neto, L.M., Barros, F., Vinhos, F., Furtado, S.G., Judice, D.M., Fernandez, E.P., Sfair, J.C., Barros, F.S.M., Prieto, P.V., Kutschenko, D.C., Moraes, M.A., Zanata, M.R.V. & Filho, L.A.F. dos S. (2013) Orchidaceae. In: Martinelli, G. & Moraes, M.A. (Eds.) Livro Vermelho da Flora do Brasil. Rio de Janeiro, pp.749–818.
Nylander, J.A.A. (2004) MrModeltest v2. Evolutionary Biology Centre, Uppsala University, Sweden. Available from: https://github.com/nylander/MrModeltest2 (accessed 5 February 2020)
Otero, J.T., Ackerman, J.D. & Bayman, P. (2002) Diversity and host specificity of endophytic Rhizoctonia-like fungi from tropical orchids. American Journal of Botany 89: 1852–1858. https://doi.org/10.3732/ajb.89.11.1852
Otero, J.T., Ackerman, J.D. & Bayman, P. (2004) Differences in mycorrhizal preferences between two tropical orchids. Molecular Ecology 13: 2393–2404. https://doi.org/10.1111/j.1365-294X.2004.02223.x
Otero, J.T., Flanagan, N.S., Herre, E.A., Ackerman, J.D. & Bayman, P. (2007) Widespread mycorrhizal specificity correlates to mycorrhizal function in the neotropical, epiphytic orchid Ionopsis utricularioides (Orchidaceae). American Journal of Botany 94 (12): 1944–1950. https://doi.org/10.3732/ajb.94.12.1944
Pereira, O.L., Rollemberg, C.L., Borges, A.C., Kasuya, M.C.M. & Matsuoka, K. (2003) Epulorhiza epiphytica sp. nov. isolated from mycorrhizal roots of epiphytic orchids in Brazil. Mycoscience 44 (2): 153–155. https://doi.org/10.1007/S10267-002-0087-7
Pereira, O.L., Kasuya, M.C.M., Borges, A.C. & Araújo, E.F. (2005) Morphological and molecular characterization of mycorrhizal fungi isolated from neotropical orchids in Brazil. Canadian Journal Botany 83: 54–65. https://doi.org/10.1139/b04-151
Pereira, M.C., Coelho, I.S., Valadares, R.B.S., Oliveira, S.F., Bocayuva, M., Pereira, O.L., Araújo, E.F. & Kasuya, M.C.M. (2014) Morphological and molecular characterization of Tulasnella spp. fungi isolated from the roots of Epidendrum secundum, a widespread Brazilian orchid. Symbiosis 62: 111–121. https://doi.org/10.1007/s13199-014-0276-0
Pereira, M.C., Pereira, O.L., Costa, M.D., Rocha, R.B. & Kasuya, M.C.M. (2009) Diversidade de fungos micorrízicos Epulorhiza spp. isolados de Epidendrum secundum (Orchidaceae). Revista Brasileira de Ciência do Solo 33: 1187–1197. https://doi.org/10.1590/S0100-06832009000500012
Nogueira, R.E., Pereira, O.L., Kasuya, M.C.M., Lanna, M.C.S. & Mendonça, M.P. (2005) Fungos micorrízicos associados a orquídeas em campos rupestres na região do Quadrilátero Ferrífero, MG, Brasil. Acta Botanica Brasilica 19 (3): 417–424. https://doi.org/10.1590/S0102-33062005000300001
Rasmussen, H.N. (2002) Recent developments in the study of orchid mycorrhiza. Plant and Soil 244 (1–2): 149–163. https://doi.org/10.1023/A:1020246715436
Ronquist, F. & Huelsenbeck, J.P. (2003) MrBayes 3: Beyesian phylogenetic inference under mixed models. Bioinformatics 19: 1572–1574. https://doi.org/10.1093/bioinformatics/btg180
Rambaut, A. (2009) FigTree version 1.4.4. Available from: http://tree.bio.ed.ac.uk/software/figtree/ (accessed 5 February 2020)
Rammitsu, K., Yagame, T., Yamashita, Y., Yukawa, T., Isshiki, S. & Ogura-Tsujita, Y. (2019) A leafless epiphytic orchid, Taeniophyllum glandulosum Blume (Orchidaceae), is specifically associated with the Ceratobasidiaceae family of basidiomycetous fungi. Mycorrhiza 29 (2): 159–166. https://doi.org/10.1007/s00572-019-00881-7
Rannala, B. & Yang, Z. (1996) Proballity distribution of molecular evolutionary trees: A new method of phylogenetic inference. Journal of Molecular Evolution 43: 304–311. https://doi.org/10.1007/BF02338839
Smith, S.E. & Read, D.J. (2008) Mycorrhizal Symbiosis, 3rd edn. Adelaide, Australia.
Thixton, H.L., Esselman, E.J., Corey, L.L. & Zettler, L.W. (2020) Further evidence of Ceratobasidium D.P. Rogers (Basidiomycota) serving as the ubiquitous fungal associate of Platanthera leucophaea (Orchidaceae) in the North American tallgrass prairie. Botanical Studies 61: 12. https://doi.org/10.1186/s40529-020-00289-z
Valadares, R.B., Pereira, M.C., Otero, J.T. & Cardoso, E.J. (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
Warcup, J.H. & Talbot, P.H.B. (1980) Perfect states of Rhizoctonias associated with orchids. New Phytologist 66 (4): 631–641. https://doi.org/10.1111/j.1469-8137.1980.tb00787.x
Weiß, M., Waller, F., Zuccano, A. & Selosse, M.A. (2016) Sebacinales – one thousand and one interactions with land plants. New Phytologist 211: 20–40. https://doi.org/10.1111/nph.13977
White, T.J., Bruns, T.D., Lee, S. & Taylor, J. (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis, M.A., Gelfand, D.H., Sninsky, J.J. & White, T.J. (Eds.) PCR protocols: a guide to methods and applications. Academic, San Diego, pp. 315–322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
Zettler, L.W., Piskin, K.A., Stewart, S.L., Hartsock, J.J., Bowles, M.L. & Bell, T.J. (2005) Protocorm mycobionts of the Federally threatened Eastern Prairie Fringed Orchid, Platanthera leucophaea (Nutt.) Lindl., and a technique to prompt leaf elongation in seedlings. Studies in Mycology 53: 163–171. https://doi.org/10.3114/sim.53.1.163