Abstract
The Serendipitaceae family was erected in 2016 to accommodate the Sebacinales ‘group B’ clade, which contains peculiar species of cultivable root–associated fungi involved in symbiotic associations with a wide range of plant species. Here we report the isolation of a new Serendipita species which was obtained from protocorms of the terrestrial orchid Epidendrum fulgens cultivated in a greenhouse. This species is described based on phylogenetic analysis and on its microscopic and ultrastructural features in pure culture and in association with the host’s protocorms. Its genome size was estimated using flow cytometry, and its capacity to promote the germination of E. fulgens seeds and to associate with roots of Arabidopsis thaliana was also investigated. Serendipita restingae sp. nov. is closely related to Serendipita sp. MAFF305841, isolated from Microtis rara (Orchidaceae), from which it differs by 14.2% in the ITS region and by 6.5% in the LSU region. It produces microsclerotia formed of non-monilioid hyphae, a feature that was not reported for the Sebacinales hitherto. Serendipita restingae promoted the germination of E. fulgens seeds, forming typical mycorrhizal pelotons within protocorm cells. It was also able to colonize the roots of Arabidopsis thaliana under in vitro conditions. Arabidopsis plants grown in association with S. restingae increased their biomass more than fourfold. Serendipita restingae is the first Serendipitaceae species described for the Americas.
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Almeida P, van den Berg C, Góes-Neto A (2014) Epulorhiza amonilioides sp. nov.: a new anamorphic species of orchid mycorrhiza from brazil. Neodiversity 7:1–10. https://doi.org/10.13102/neod.71.1
Araujo DSD (1992) Vegetation types of sandy coastal plains of tropical Brazil: A first approximation. In: Seeliger U (ed) Coastal plant communities of Latin America, 1st edn. Academic Press, San Diego, pp 337–347
Basiewicz M, Weiß M, Kogel KH et al (2012) Molecular and phenotypic characterization of Sebacina vermifera strains associated with orchids, and the description of Piriformospora williamsii sp. nov. Fungal Biol 116:204–213. https://doi.org/10.1016/j.funbio.2011.11.003
Bennett MD, Leitch IJ, Price HJ, Johnston JS (2003) Comparisons with Caenorhabditis (∼100 Mb) and Drosophila (∼175 Mb) using flow cytometry show genome size in Arabidopsis to be ∼157 Mb and thus ∼25% larger than the Arabidopsis genome initiative estimate of ∼125 Mb. Ann Bot 91:547–557. https://doi.org/10.1093/aob/mcg057
Berch SM, Allen TR, Berbee ML (2002) Molecular detection, community structure and phylogeny of ericoid mycorrhizal fungi. Plant Soil 244:55–66. https://doi.org/10.1023/A:1020291516884
Darriba D, Taboada GL, Doallo R, Posada D (2012) jModelTest 2: more models, new heuristics and parallel computing CircadiOmics: integrating circadian genomics, transcriptomics, proteomics. Nat Methods 9:772. https://doi.org/10.1038/nmeth.2109
Dearnaley JDW, Perotto S, Selosse M-A (2017) Structure and development of orchid mycorrhizas. In: Martin F (ed) Molecular Mycorrhizal Symbiosis, 1st edn. John Wiley & Sons, New Jersey, pp 63–86
Deshmukh S, Hückelhoven R, Schäfer P et al (2006) The root endophytic fungus Piriformospora indica requires host cell for proliferation during mutualistic symbiosis with barley. Proc Natl Acad Sci U S A 103:18450–18457. https://doi.org/10.1073/pnas.0605697103
Doležel J, Bartoš J (2005) Plant DNA flow cytometry and estimation of nuclear genome size. Ann Bot 95:99–110. https://doi.org/10.1093/aob/mci005
Doležel J, Bartos J, Voglmayr H, Greilhuber J (2003) Nuclear DNA content and genome size of trout and human. Cytometry A 51:127–128; author reply 129. doi: https://doi.org/10.1002/cyto.a.10013
Doyle JJ, Doyle J (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15
Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792–1797. https://doi.org/10.1093/nar/gkh340
Galbraith DW, Harkins KR, Maddox JM et al (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220(4601):1049–1051. https://doi.org/10.1016/b978-0-12-715001-7.50090-1
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. https://doi.org/10.1111/j.1365-294X.1993.tb00005.x
Garnica S, Riess K, Schön ME et al (2016) Divergence times and phylogenetic patterns of Sebacinales, a highly diverse and widespread fungal lineage. PLoS ONE 11:1–16. https://doi.org/10.1371/journal.pone.0149531
Ghimire SR, Craven KD (2011) Enhancement of switchgrass (Panicum virgatum L.) biomass production under drought conditions by the ectomycorrhizal fungus Sebacina vermifera. Appl Environ Microbiol 77:7063–7067. https://doi.org/10.1128/AEM.05225-11
Girlanda M, Segreto R, Cafasso D et al (2011) Photosynthetic Mediterranean meadow orchids feature partial mycoheterotrophy and specific mycorrhizal associations1. Am J Bot 98:1148–1163. https://doi.org/10.3732/ajb.1000486
Hashimoto Y, Fukukawa S, Kunishi A et al (2012) Mycoheterotrophic germination of Pyrola asarifolia dust seeds reveals convergences with germination in orchids. New Phytol 195:620–630. https://doi.org/10.1111/j.1469-8137.2012.04174.x
He M, Zhao R, Hyde KD et al (2019) Notes, outline and divergence times of Basidiomycota. Fungal Divers 4:1–263. https://doi.org/10.1007/s13225-019-00435-4
Jeewon R, Hyde KD (2016) Establishing species boundaries and new taxa among fungi: recommendations to resolve taxonomic ambiguities. Mycosphere 7:1669–1677. https://doi.org/10.5943/mycosphere/7/11/4
Johnson JM, Sherameti I, Ludwig A et al (2011) Protocols for Arabidopsis thaliana and Piriformospora indica co-cultivation – a model system to study plant beneficial traits. J Endocytobiosis Cell Res 21:101–113
Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J Cell Biol 27:137-138A
Kirk PM, Stalpers JA, Braun U et al (2013) A without-prejudice list of generic names of fungi for protection under the International Code of Nomenclature for algae, fungi, and plants. IMA Fungus 4:381–443. https://doi.org/10.5598/imafungus.2013.04.02.17
Kohler A, Kuo A, Nagy LG et al (2015) Convergent losses of decay mechanisms and rapid turnover of symbiosis genes in mycorrhizal mutualists. Nat Genet 47:410–415. https://doi.org/10.1038/ng.3223
Kottke I, Beiter A, Weiß M et al (2003) Heterobasidiomycetes form symbiotic associations with hepatics: Jungermanniales have sebacinoid mycobionts while Aneura pinguis (Metzgeriales) is associated with a Tulasnella species. Mycol Res 107:957–968. https://doi.org/10.1017/S0953756203008141
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 202:594–605. https://doi.org/10.1111/nph.12700
Lacercat-Didier L, Berthelot C, Foulon J et al (2016) New mutualistic fungal endophytes isolated from poplar roots display high metal tolerance. Mycorrhiza 26:657–671. https://doi.org/10.1007/s00572-016-0699-y
Lahrmann U, Zuccaro A (2012) Opprimo ergo sum-evasion and suppression in the root endophytic fungus Piriformospora indica. Mol Plant-Microbe Interact 25:727–737. https://doi.org/10.1094/MPMI-11-11-0291
Milligan MJ, Williams PG (1987) Orchidaceous rhizoctonias from roots of nonorchids: mycelial and cultural characteristics of field and pot culture isolates. Can J Bot 65:598–606. https://doi.org/10.1139/b87-077
Milligan MJ, Williams PG (1988) The mycorrhizal relationship of multinucleate rhizoctonias from non-orchids with Microtis (Orchidaceae). New Phytol 108:205–209. https://doi.org/10.1111/j.1469-8137.1988.tb03697.x
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:474–497
Novotná A, Suárez JP (2018) Molecular detection of bacteria associated with Serendipita sp., a mycorrhizal fungus from the orchid Stanhopea connata Klotzsch in southern Ecuador. Bot Lett 165:307–313. https://doi.org/10.1080/23818107.2018.1436087
O’Donnell K (1993) Fusarium and its Near Relatives. In: Reynolds DR, Taylor JW (eds) The fungal holomorph: mitotic, meiotic and pleomorphic speciation in fungal systematics, 1st edn. CAB International, Wallingford, pp 225–233
Oberwinkler F (1964) Intrahymeniale Heterobasidiomyceten. Fruchtkorperlose Sebacina-Sippen und ihre systematische stellung. Nova Hedwigia 7:489–499
Oberwinkler F, Riess K, Bauer R et al (2013) Enigmatic Sebacinales Mycol Prog 12:1–27. https://doi.org/10.1007/s11557-012-0880-4
Oberwinkler F, Riess K, Bauer R, Garnica S (2014) Morphology and molecules: the Sebacinales, a case study. Mycol Prog 13:445–470. https://doi.org/10.1007/s11557-014-0983-1
Oliveira SF, Bocayuva MF, Veloso TGR et al (2014) Endophytic and mycorrhizal fungi associated with roots of endangered native orchids from the Atlantic Forest, Brazil. Mycorrhiza 24:55–64. https://doi.org/10.1007/s00572-013-0512-0
Otero JT, Ackerman JD, Bayman P (2004) Differences in mycorrhizal preferences between two tropical orchids. Mol Ecol 13(8):2393–2404
Otto F (1990) DAPI Staining of fixed cells for high-resolution flow cytometry of nuclear DNA. In: Crissman HA, Darzinskievikcz Z (eds) Methods in cell biology. Academic Press, New York, pp 105–110
Peškan-Berghöfer T, Shahollari B, Pham HG et al (2004) Association of Piriformospora indica with Arabidopsis thaliana roots represents a novel system to study beneficial plant-microbe interactions and involves early plant protein modifications in the endoplasmic reticulum and at the plasma membrane. Physiol Plant 122:465–477. https://doi.org/10.1111/j.1399-3054.2004.00424.x
R Core Team (2019). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. https://www.Rproject.org/
Rasmussen HN (1995) Terrestrial orchids: from seed to mycotrophic plant. Cambridge University Press, Cambridge
Rasmussen HN (2002) Recent developments in the study of orchid mycorrhiza. Plant Soil 244:149–163
Rasmussen HN, Dixon KW, Jersáková J, Těšitelová T (2015) Germination and seedling establishment in orchids: a complex of requirements. Ann Bot 116:391–402. https://doi.org/10.1093/aob/mcv087
Ray P, Craven KD (2016) Sebacina vermifera: a unique root symbiont with vast agronomic potential. World J Microbiol Biotechnol 32:1–10. https://doi.org/10.1007/s11274-015-1970-7
Ray P, Ishiga T, Decker SR et al (2015) A novel delivery system for the root symbiotic fungus, Sebacina vermifera, and consequent biomass enhancement of low lignin COMT switchgrass lines. Bioenergy Res 8:922–933. https://doi.org/10.1007/s12155-015-9636-8
Riess K, Oberwinkler F, Bauer R, Garnica S (2014) Communities of endophytic sebacinales associated with roots of herbaceous plants in agricultural and grassland ecosystems are dominated by Serendipita herbamans sp. nov. PLoS One 9:e94676 https://doi.org/10.1371/journal.pone.0094676
Roberts P (1993) Exidiopsis species from Devon, including the new segregate genera Ceratosebacina, Endoperplexa, Microsebacina, and Serendipita. Mycol Res 97:467–478. https://doi.org/10.1016/S0953-7562(09)80135-4
Ronquist F, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19:1572–1574. https://doi.org/10.1093/bioinformatics/btg180
Scarano FR (2002) Structure, function and floristic relationships of plant communities in stressful habitats marginal to the Brazilian Atlantic rainforest. Ann Bot 90:517–524. https://doi.org/10.1093/aob/mcf189
Schoch CL, Seifert KA, Huhndorf S et al (2012) Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proc Natl Acad Sci 109:6241–6246. https://doi.org/10.1073/pnas.1117018109
Selosse M-A (2014) The latest news from biological interactions in orchids: in love, head to toe. New Phytol 202:337–340. https://doi.org/10.1111/nph.12769
Selosse M-A, Dubois MP, Alvarez N (2009) Do Sebacinales commonly associate with plant roots as endophytes? Mycol Res 113:1062–1069. https://doi.org/10.1016/j.mycres.2009.07.004
Selosse M-A, Martos F (2014) Do chlorophyllous orchids heterotrophically use mycorrhizal fungal carbon? Trends Plant Sci 19:683–685. https://doi.org/10.1016/j.tplants.2014.09.005
Selosse M-A, Setaro SD, Glatard F et al (2007) Sebacinales are common mycorrhizal associates of Ericaceae. New Phytol 174:864–878. https://doi.org/10.1111/j.1469-8137.2007.02064.x
Setaro SD, Garnica S, Herrera PI et al (2011) A clustering optimization strategy to estimate species richness of Sebacinales in the tropical Andes based on molecular sequences from distinct DNA regions. Biodivers Conserv 21:2269–2285. https://doi.org/10.1007/s10531-011-0205-y
Setaro SD, Kron K (2011) Neotropical and north American Vaccinioideae (Ericaceae) share their mycorrhizal Sebacinales-an indication for concerted migration? PLoS Curr 3:1–19. https://doi.org/10.1371/currents.RRN1227
Setaro SD, Weiß M, Oberwinkler F, Kottke I (2006) Sebacinales form ectendomycorrhizas with Cavendishia nobilis, a member of the Andean clade of Ericaceae, in the mountain rain forest of southern Ecuador. New Phytol 169:355–365. https://doi.org/10.1111/j.1469-8137.2005.01583.x
Stamatakis A (2014) RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30:1312–1313. https://doi.org/10.1093/bioinformatics/btu033
Suárez JP, Weiß M, Abele A, et al (2008) Members of Sebacinales subgroup B form mycorrhizae with epiphytic orchids in a neotropical mountain rain forest. Mycol Progress 7:75. https://doi.org/10.1007/s11557-008-0554-4
Suárez JP, Weiß M, Abele A et al (2006) Diverse tulasnelloid fungi form mycorrhizas with epiphytic orchids in an Andean cloud forest. Mycol Res 110:1257–1270. https://doi.org/10.1016/j.mycres.2006.08.004
Suárez JP, Weiß M, Oberwinkler F, Kottke I (2009) Epiphytic orchids in a mountain rain forest in southern Ecuador harbor groups of mycorrhiza- forming Tulasnellales and Sebacinales subgroup b (Basidiomycota). Proc Second Sci Conf Andean Orchid 184–196.
Sujii PS, Cozzolino S, Pinheiro F (2019) Hybridization and geographic distribution shapes the spatial genetic structure of two co-occurring orchid species. Heredity 123:458–469. https://doi.org/10.1038/s41437-019-0254-7
Tavares S, Ramos AP, Pires AS et al (2014) Genome size analyses of Pucciniales reveal the largest fungal genomes. Front Plant Sci. https://doi.org/10.3389/fpls.2014.00422
Townsend BB, Willetts HJ (1954) The development of sclerotia of certain fungi. Trans Br Mycol Soc 37:213–221. https://doi.org/10.1016/s0007-1536(54)80003-9
Venneman J, Audenaert K, Verwaeren J et al (2017) Congolese rhizospheric soils as a rich source of new plant growth-promoting endophytic Piriformospora isolates. Front Microbiol 8:1–15. https://doi.org/10.3389/fmicb.2017.00212
Verma S, Varma A, Rexer K et al (1998) Piriformospora indica, gen. et sp. nov., a new root-colonizing fungus. Mycologia 90:896–903
Voges JG, Benevenuto R, Fritsche Y et al (2014) Protocorm development of Epidendrum fulgens (Orchidaceae) in response to different saline formulations and culture conditions. Acta Sci Biol Sci 36(3):287–292
Vohník M, Pánek M, Fehrer J, Selosse M-A (2016) Experimental evidence of ericoid mycorrhizal potential within Serendipitaceae (Sebacinales). Mycorrhiza 26:831–846. https://doi.org/10.1007/s00572-016-0717-0
Warcup JH (1981) The mycorrhizal relationships of Australian orchids. New Phytol 87:371–381. https://doi.org/10.1111/j.1469-8137.1981.tb03208.x
Warcup JH (1988) Mycorrhizal associations of isolates of Sebacina vermifera. New Phytol 110:227–231. https://doi.org/10.1111/j.1469-8137.1988.tb00256.x
Warcup JH, Talbot PHB (1967) Perfect states of Rhizoctonias associated with orchids. New Phytol 66:631–641
Weiß M, Selosse M-A, Rexer K-H et al (2004) Sebacinales: a hitherto overlooked cosm of heterobasidiomycetes with a broad mycorrhizal potential. Mycol Res 108:1003–1010. https://doi.org/10.1017/S0953756204000772
Weiß M, Sýkorová Z, Garnica S et al (2011) Sebacinales everywhere: previously overlooked ubiquitous fungal endophytes. PLoS ONE 6:1–8. https://doi.org/10.1371/journal.pone.0016793
Weiß M, Waller F, Zuccaro A, Selosse M-A (2016) Sebacinales — one thousand and one interactions with land plants. New Phytol 211:20–40. https://doi.org/10.1111/NPH.13977
Williams PG, Thilo E (1989) Ultrastructural evidence for the identity of some multinucleate rhizoctonias. New Phytol 112:513–518. https://doi.org/10.1111/j.1469-8137.1989.tb00345.x
Wilson AD (1992) A versatile giemsa protocol for permanent nuclear staining of fungi. Mycologia 84:585–588. https://doi.org/10.1080/00275514.1992.12026181
Wilson D (1995) Endophyte: the evolution of a term, and clarification of its use and definition. Nord Soc Oikos 73:274–276. https://doi.org/10.2307/3545919
Yagame T, Yamato M (2008) Isolation and identification of mycorrhizal fungi associated with Stigmatodactylus sikokianus (Maxim. ex Makino) Rauschert (Orchidaceae). Mycoscience 49:388–391. https://doi.org/10.1007/s10267-008-0436-2
Zuccaro A, Basiewicz M, Zurawska M et al (2009) Karyotype analysis, genome organization, and stable genetic transformation of the root colonizing fungus Piriformospora indica. Fungal Genet Biol 46:543–550. https://doi.org/10.1016/j.fgb.2009.03.009
Zuccaro A, Lahrmann U, Güldener U et al (2011) Endophytic life strategies decoded by genome and transcriptome analyses of the mutualistic root symbiont Piriformospora indica. PLoS Pathog. https://doi.org/10.1371/journal.ppat.1002290
White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and Direct Sequencing of Fungal Ribosomal Rna Genes for Phylogenetics. In: MA I, DHG, JJ S, TJ W (eds) PCR Protocols: a Guide to Methods and Applications, 1st edn. Academic Press, New York., pp 315–322
Smith ME, Henkel TW, Rollins JA (2015) How many fungi make sclerotia? Fungal Ecol 13:211–220. https://doi.org/10.1016/j.funeco.2014.08.010
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Fritsche, Y., Lopes, M.E., Selosse, MA. et al. Serendipita restingae sp. nov. (Sebacinales): an orchid mycorrhizal agaricomycete with wide host range. Mycorrhiza 31, 1–15 (2021). https://doi.org/10.1007/s00572-020-01000-7
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DOI: https://doi.org/10.1007/s00572-020-01000-7