Gibellula aurea sp. nov. (Ascomycota, Cordycipitaceae): a new golden spider-devouring fungus from a Brazilian Atlantic Rainforest

THAIRINE MENDES-PEREIRA; JOÃO PAULO MACHADO DE ARAÚJO; FLÁVIA CARVALHO MENDES; EMILY OLIVEIRA FONSECA; JULIE ERICA DA ROCHA ALVES; JOBER FERNANDO SOBCZAK; ARISTÓTELES GÓES-NETO

doi:10.11646/phytotaxa.573.1.5

Issue: Vol. 573 No. 1: 17 November 2022

Type: Article

Published: 2022-11-17

DOI: 10.11646/phytotaxa.573.1.5

Page range: 85-102

Abstract views: 203

PDF downloaded: 3

Gibellula aurea sp. nov. (Ascomycota, Cordycipitaceae): a new golden spider-devouring fungus from a Brazilian Atlantic Rainforest

THAIRINE MENDES-PEREIRA⁺⁻
JOÃO PAULO MACHADO DE ARAÚJO⁺⁻
FLÁVIA CARVALHO MENDES⁺⁻
EMILY OLIVEIRA FONSECA⁺⁻
JULIE ERICA DA ROCHA ALVES⁺⁻
JOBER FERNANDO SOBCZAK⁺⁻
ARISTÓTELES GÓES-NETO⁺⁻

Laboratory of Molecular and Computational Biology of Fungi, Department of Microbiology, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG, Brazil

Institute of Systematic Botany, The New York Botanical Garden, 2900 Southern Blvd., Bronx, NY 10458-5126, U.S.A.

Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG, Brazil

Program in Ecology and Natural Resources, Department of Biology, Universidade Federal do Ceará, 60440-900, Fortaleza, CE, Brazil

Laboratory of Ecology and Evolution, Institute of Exact Sciences and Nature; Universidade Internacional da Lusofonia Afro-Brasileira, 62790-000, Redenção, CE, Brazil

Program in Ecology and Natural Resources, Department of Biology, Universidade Federal do Ceará, 60440-900, Fortaleza, CE, Brazil; Laboratory of Ecology and Evolution, Institute of Exact Sciences and Nature; Universidade Internacional da Lusofonia Afro-Brasileira, 62790-000, Redenção, CE, Brazil

Laboratory of Molecular and Computational Biology of Fungi, Department of Microbiology, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG, Brazil

Behavioural manipulation host-parasite interaction Hypocreales multigene phylogeny spider pathogen taxonomy Fungi

Requires Subscription or Fee PDF (7MB)

Abstract

Fungal parasites that infect spiders can affect their survival and potentially their behaviour. The genus Gibellula (Ascomycota, Cordycipitaceae) infects arachnids, and due to difficulties in identifying hosts after fungal sporulation, its ecology and spider host diversity are poorly understood. Herein, we propose a new species of Gibellula from the Brazilian Atlantic Forest: Gibellula aurea, which parasitizes two spider families (Anyphaenidae and Corinnidae). Gibellula aurea exhibits characteristic golden-yellow hyphae that completely cover hosts and white conidiophores emerging along synnemata or directly from hosts. Molecular identification was performed by sequencing the small and large nuclear ribosomal subunits, translation elongation factor, and RNA polymerase II second largest subunit regions, and reconstructing multigene phylogenetic trees. Maximum Likelihood and Bayesian Inference phylogenies demonstrate that G. aurea forms a well-supported clade closer to G. flava, G. gamsii, G. leiopus, G. longispora, G. pigmentosinum, and G. pulchra. The phylogenetic reconstruction displayed herein represents the most comprehensive phylogeny for this genus to date. Spider hosts parasitized by G. aurea have been invariably found attached underneath leaves in vegetation, regardless of the identity of the infected hosts. The consistency of the death locations of G. aurea hosts indicates recurrent behaviour given the habitat heterogeneity of these host groups. Nevertheless, it is still unclear to what extent the place where hosts die can favour parasite fitness.

References

Alizon, S., de Roode, J.C. & Michalakis, Y. (2013) Multiple infections and the evolution of virulence. Ecology Letters 16: 556–567. https://doi.org/10.1111/ele.12076
Araújo, J.P.M., Evans, H.C., Geiser, D.M., Mackay, W.P. & Hughes, D.P. (2015) Unravelling the diversity behind the Ophiocordyceps unilateralis (Ophiocordycipitaceae) complex: Three new species of zombie-ant fungi from the Brazilian Amazon. Phytotaxa 220: 224–238. https://doi.org/10.11646/phytotaxa.220.3.2
Araújo, J.P.M., Evans, H.C., Kepler, R. & Hughes, D.P. (2018) Zombie-ant fungi across continents: 15 new species and new combinations within Ophiocordyceps. I. Myrmecophilous hirsutelloid species. Studies in Mycology 90: 119–160. https://doi.org/10.1016/j.simyco.2017.12.002 
Araújo, J.P.M. & Hughes, D.P. (2016) Diversity of entomopathogenic fungi: which groups conquered the insect body? Advances in Genetics. Elsevier, Academic Press, pp. 1–39. 
Araújo, J.P.M. & Hughes, D.P. (2019) Zombie-ant fungi emerged from non-manipulating, beetle-infecting ancestors. Current Biology 29: 3735–3738. https://doi.org/10.1016/j.cub.2019.09.004 
Arruda, I.D.P., Villanueva-Bonilla, G.A., Faustino, M.L., Moura-Sobczak, J.C.M.S. & Sobczak, J.F. (2021) Behavioral manipulation of the spider Macrophyes pacoti (Araneae: Anyphaenidae) by the araneopathogenic fungus Gibellula sp. (Hypocreales: Cordycipitaceae). Canadian Journal of Zoology 99: 401–408. https://doi.org/10.1139/cjz-2020-0232 
Bétard, F., Peulvast, J.P. & Claudino-Sales, V. (2007) Caracterização morfopedólogica de uma serra úmida no semi-árido do nordeste brasileiro: o caso do Maciço de Baturité-CE (morphopedological characterization of a humid mountain in the brazilian semi-arid north-east). Mercator, Fortaleza, v. 6, n. 12, pp. 107 a 126, nov. 2008. ISSN 1984-2201. Available from: http://www.mercator.ufc.br/mercator/article/view/49 (accessed 16 November 2022)
Brescovit, A.D., Villanueva-Bonilla, G.A., Sobczak, J.C.M.S.M., Nóbrega, F.A.D.S., Oliveira, L.F.M., Arruda, I.D.P. & Sobczak, J.F. (2019) Macrophyes pacoti n. sp. (Araneae: Anyphaenidae) from Brazilian Atlantic Forest, with notes on an araneopathogenic fungus. Zootaxa 4629: 294–300. https://doi.org/10.11646/zootaxa.4629.2.12 
Castlebury, L.A., Rossman, A.Y., Sung, G.-H., Hyten, A.S. & Spatafora, J.W. (2004) Multigene phylogeny reveals new lineage for Stachybotrys chartarum, the indoor air fungus. Mycological Research 108: 864–872. https://doi.org/10.1017/S0953756204000607 
Cavara, F. (1894) Ulteriore contribuzione alla micologia lombarda. In: Atti dell’Istituto Botanico della Università e Laboratorio Crittogamico di Pavia. pp. 313–150. 
Chen, M., Wang, T., Lin, Y. & Huang, B. (2022) Morphological and molecular analyses reveal two new species of Gibellula (Cordycipitaceae, Hypocreales) from China. MycoKeys 90: 53–69. https://doi.org/10.3897/mycokeys.90.83801 
Chen, M.-J., Wang, T., Lin, Y. & Huang, B. (2021) Gibellula flava sp. nov. (Cordycipitaceae, Hypocreales), a new pathogen of spider from China. Phytotaxa 527: 125–133. https://doi.org/10.11646/phytotaxa.527.2.5 
Chen, W.-H., Han, Y.-F., Liang, Z.-Q. & Zou, X. (2016) Morphological traits, DELTA system, and molecular analysis for Gibellula clavispora sp. nov. from China. Mycotaxon 131: 111–121. https://doi.org/10.5248/131.111
Clancy, L.M., Jones, R., Cooper, A.L., Griffith, G.W. & Santer, R.D. (2018) Dose-dependent behavioural fever responses in desert locusts challenged with the entomopathogenic fungus Metarhizium acridum. Scientific Reports 8: 14222. https://doi.org/10.1038/s41598-018-32524-w 
Costa, P.P. (2014) Gibellula spp. associadas a aranhas da Mata do Paraíso, Viçosa-MG (M.Sc.). Universidade Federal de Viçosa. Minas Gerais, Brasil.
Elliot, S.L., Blanford, S. & Thomas, M.B. (2002) Host–pathogen interactions in a varying environment: temperature, behavioural fever and fitness. Proceedings of the Royal Society of London. Series B: Biological Sciences 269: 1599–1607. https://doi.org/10.1098/rspb.2002.2067 
Evans, H.C. (1974) Natural control of arthropods, with special reference to ants (Formicidae), by fungi in the tropical high forest of Ghana. The Journal of Applied Ecology 11: 37–49. https://doi.org/10.2307/2402003 
Evans, H.C. (1988) Coevolution of entomogenous fungi and their insect hosts. In: Pirozynski, K.A. & Hawksworth, D.L. (eds.) Coevolution of fungi with plants and animals. Academic Press Limited, San Diego, USA, pp. 149–172. 
Evans, H.C. (2013) Fungal Pathogens of Spiders. In: Nentwig, W. (ed.) Spider Ecophysiology. Springer Berlin Heidelberg, Berlin, Heidelberg, pp. 107–121. 
Evans, H.C., Elliot, S.L. & Hughes, D.P. (2011) Hidden diversity behind the zombie-ant fungus Ophiocordyceps unilateralis: four new species described from carpenter ants in Minas Gerais, Brazil. PLoS ONE 6: e17024. https://doi.org/10.1371/journal.pone.0017024 
Geneious Prime (2021) Geneious Prime ® 2021.2.2. Available from: https://www.geneious.com (accessed 16 November 2022)
Höfer, H. & Brescovit, A.D. (2001) Species and guild structure of a Neotropical spider assemblage (Araneae) from Reserva Ducke, Amazonas, Brazil. Andrias 15: 99–119. 
Hubert, N. & Hanner, R. (2015) DNA Barcoding, species delineation and taxonomy: a historical perspective. DNA Barcodes 3: 44–58. https://doi.org/10.1515/dna-2015-0006 
Hughes, D.P., Araújo, J.P.M., Loreto, R.G., Quevillon, L., de Bekker, C. & Evans, H.C. (2016) From So Simple a Beginning: the evolution of behavioral manipulation by fungi. Advances in Genetics. Elsevier, pp. 437–469. 
Index Fungorum (2022) Index Fungorum. Available from: www.indexfungorum.org (accessed 30 May 2022)
Inkscape (2021) Inkscape: Draw freely. Available from: https://inkscape.org (accessed on 10 July 2022)
Jocqué, R. & Dippenaar-Schoeman, A.S. (2007) Spider families of the world. 2nd ed. Musée Royal de l’Afrique Centrale, Tervuren, Belgium. 336 pp. 
Johnson, D., Sung, G.-H., Hywel-Jones, N.L., Luangsa-Ard, J.J., Bischoff, J.F., Kepler, R.M. & Spatafora, J.W. (2009) Systematics and evolution of the genus Torrubiella (Hypocreales, Ascomycota). Mycological Research 113: 279–289. https://doi.org/10.1016/j.mycres.2008.09.008 
Kepler, R.M., Kaitsu, Y., Tanaka, E., Shimano, S. & Spatafora, J.W. (2011) Ophiocordyceps pulvinata sp. nov., a pathogen with a reduced stroma. Mycoscience 52: 39–47. https://doi.org/10.1007/S10267-010-0072-5 
Kepler, R.M., Luangsa-Ard, J.J., Hywel-Jones, N.L., Quandt, C.A., Sung, G.-H., Rehner, S.A., Aime, M.C., Henkel, T.W., Sanjuan, T., Zare, R., Chen, M., Li, Z., Rossman, A.Y., Spatafora, J.W. & Shrestha, B. (2017) A phylogenetically-based nomenclature for Cordycipitaceae (Hypocreales). IMA fungus 8: 335–353. https://doi.org/10.5598/imafungus.2017.08.02.08 
Kobayasi, Y. & Shimizu, D. (1977) Some species of Cordyceps and its allies on spiders. Kew Bulletin 31: 557–566. https://doi.org/10.2307/4119402 
Kolde, R. (2019) pheatmap: Pretty Heatmaps. R package version 1.0.12. R Core Team. 
Kozlov, A.M., Darriba, D., Flouri, T., Morel, B. & Stamatakis, A. (2019) RAxML-NG: a fast, scalable and user-friendly tool for maximum likelihood phylogenetic inference. Bioinformatics (Oxford, England) 35: 4453–4455. https://doi.org/10.1093/bioinformatics/btz305 
Kuephadungphan, W., Macabeo, A.P.G., Luangsa-ard, J.J., Tasanathai, K., Thanakitpipattana, D., Phongpaichit, S., Yuyama, K. & Stadler, M. (2019) Studies on the biologically active secondary metabolites of the new spider parasitic fungus Gibellula gamsii. Mycological Progress 18: 135–146. https://doi.org/10.1007/s11557-018-1431-4 
Kuephadungphan, W., Petcharad, B., Tasanathai, K., Thanakitpipattana, D., Kobmoo, N., Khonsanit, A., Samson, R.A. & Luangsa-ard, J.J. (2022) Multi-locus phylogeny unmasks hidden species within the specialised spider-parasitic fungus, Gibellula (Hypocreales, Cordycipitaceae) in Thailand. Studies in Mycology 101 (1): 245–286. https://doi.org/10.3114/sim.2022.101.04 
Kuephadungphan, W., Tasanathai, K., Petcharad, B., Khonsanit, A., Stadler, M. & Luangsa-ard, J.J. (2020) Phylogeny- and morphology-based recognition of new species in the spider-parasitic genus Gibellula (Hypocreales, Cordycipitaceae) from Thailand. MycoKeys 72: 17–42. https://doi.org/10.3897/mycokeys.72.55088 
Largent, D.L., Johnson, D. & Watling, R. (1977) How to identify mushrooms to genus. III: Microscopic features. Mad River, Eureka, California.
Li, Y., Steenwyk, J.L., Chang, Y., Wang, Y., James, T.Y., Stajich, J.E., Spatafora, J.W., Groenewald, M., Dunn, C.W., Hittinger, C.T., Shen, X.-X. & Rokas, A. (2021) A genome-scale phylogeny of the Kingdom Fungi. Current Biology 31: 1653–1665.e5. https://doi.org/10.1016/j.cub.2021.01.074 
Liu, Y.J., Whelen, S. & Hall, B.D. (1999) Phylogenetic relationships among Ascomycetes: evidence from an RNA polymerse II subunit. Molecular Biology and Evolution 16: 1799–1808. https://doi.org/10.1093/oxfordjournals.molbev.a026092 
Luangsa-Ard, J.J., Ridkaew, R., Tasanathai, K., Thanakitpipattana, D. & Hywel-Jones, N. (2011) Ophiocordyceps halabalaensis: a new species of Ophiocordyceps pathogenic to Camponotus gigas in Hala Bala Wildlife Sanctuary, Southern Thailand. Fungal Biology 115: 608–614. https://doi.org/10.1016/j.funbio.2011.03.002 
Mains, E.B. (1950) The Genus Gibellula on Spiders in North America. Mycologia 42: 306–321. https://doi.org/10.1080/00275514.1950.12017836 
Miller, M.A., Pfeiffer, W. & Schwartz, T. (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic trees. In: 2010 Gateway Computing Environments Workshop (GCE). IEEE, New Orleans, LA, USA, pp. 1–8. 
NCBI Resource Coordinators, Agarwala, R., Barrett, T., Beck, J., Benson, D.A., Bollin, C., Bolton, E., Bourexis, D., Brister, J.R., Bryant, S.H., Canese, K., Cavanaugh, M., Charowhas, C., Clark, K., Dondoshansky, I., Feolo, M., Fitzpatrick, L., Funk, K., Geer, L.Y., Gorelenkov, V., Graeff, A., Hlavina, W., Holmes, B., Johnson, M., Kattman, B., Khotomlianski, V., Kimchi, A., Kimelman, M., Kimura, M., Kitts, P., Klimke, W., Kotliarov, A., Krasnov, S., Kuznetsov, A., Landrum, M.J., Landsman, D., Lathrop, S., Lee, J.M., Leubsdorf, C., Lu, Z., Madden, T.L., Marchler-Bauer, A., Malheiro, A., Meric, P., Karsch-Mizrachi, I., Mnev, A., Murphy, T., Orris, R., Ostell, J., O’Sullivan, C., Palanigobu, V., Panchenko, A.R., Phan, L., Pierov, B., Pruitt, K.D., Rodarmer, K., Sayers, E.W., Schneider, V., Schoch, C.L., Schuler, G.D., Sherry, S.T., Siyan, K., Soboleva, A., Soussov, V., Starchenko, G., Tatusova, T.A., Thibaud-Nissen, F., Todorov, K., Trawick, B.W., Vakatov, D., Ward, M., Yaschenko, E., Zasypkin, A. & Zbicz, K. (2018) Database resources of the National Center for Biotechnology Information. Nucleic Acids Research 46: D8–D13. https://doi.org/10.1093/nar/gkx1095 
Nilsson, R.H., Larsson, K.-H., Taylor, A.F.S., Bengtsson-Palme, J., Jeppesen, T.S., Schigel, D., Kennedy, P., Picard, K., Glöckner, F.O., Tedersoo, L., Saar, I., Kõljalg, U. & Abarenkov, K. (2019) The UNITE database for molecular identification of fungi: handling dark taxa and parallel taxonomic classifications. Nucleic Acids Research 47: D259–D264. https://doi.org/10.1093/nar/gky1022
Nylander, J.A.A. (2004) MrModeltest v2. Program distributed by the author. Evolutionary Biology Centre, Uppsala University. 
Oliveira, T.S. de & Araújo, F.S. (2007) Diversidade e conservação da biota na Serra de Baturité, Ceará. Edições UFC; COELCE, Fortaleza. 465 pp. 
Petch, T. (1932) A list of the entomogenous fungi of Great Britain. Transactions of the British Mycological Society 17: 170–178. 
Pinheiro, J., Bates, D., DebRoy, S., Sarkar, D. & R Core Team (2021) nlme: Linear and Nonlinear Mixed Effects Models. R package version 3.1-152. R Core Team.
Pontoppidan, M.-B., Himaman, W., Hywel-Jones, N.L., Boomsma, J.J. & Hughes, D.P. (2009) Graveyards on the move: the spatio-temporal distribution of dead Ophiocordyceps-infected ants. PLoS ONE 4: e4835. https://doi.org/10.1371/journal.pone.0004835 
Poulin, R. (1995) “Adaptive” changes in the behaviour of parasitized animals: A critical review. International Journal for Parasitology 25: 1371–1383. https://doi.org/10.1016/0020-7519(95)00100-X 
Poulin, R. (2010) Parasite manipulation of host behavior: an update and frequently asked questions. In: Advances in the Study of Behavior. Elsevier, Academic Press, pp. 151–186. 
Puillandre, N., Lambert, A., Brouillet, S. & Achaz, G. (2012) ABGD, Automatic barcode gap discovery for primary species delimitation. Molecular Ecology 21: 1864–1877. https://doi.org/10.1111/j.1365-294X.2011.05239.x 
Qasim, M., Baohua, W., Zou, H., Lin, Y., Dash, C.K., Bamisile, B.S., Hussain, M., Zhiwen, Z. & Wang, L. (2018a) Phylogenetic relationship and genetic diversity of citrus psyllid populations from China and Pakistan and their associated Candidatus bacterium. Molecular Phylogenetics and Evolution 126: 173–180. https://doi.org/10.1016/j.ympev.2018.04.028 
Qasim, M., Lin, Y., Dash, C.K., Bamisile, B.S., Ravindran, K., Islam, S.U., Ali, H., Wang, F. & Wang, L. (2018b) Temperature-dependent development of Asian citrus psyllid on various hosts, and mortality by two strains of Isaria. Microbial Pathogenesis 119: 109–118. https://doi.org/10.1016/j.micpath.2018.04.019
R Core Team (2022) R: A language and environment for statistical computing. R Core Team, Vienna, Austria. 
Rambaut, A. (2017) FigTree: tree figure drawing tool version 1.4. 3. 2009. Available from: http://tree.bio.ed.ac.uk/software/figtree/ (accessed 10 July 2022).
Redman, E.M., Wilson, K. & Cory, J.S. (2016) Trade-offs and mixed infections in an obligate-killing insect pathogen. Journal of Animal Ecology 85: 1200–1209. https://doi.org/10.1111/1365-2656.12547 
Romero, G.Q. & Vasconcellos-Neto, J. (2007) Aranhas sobre plantas: dos comportamentos de forrageamento às associações específicas. In: Gonzaga, M.O., Santos, A.J. & Japyassú, H.F. (eds.) Ecologia e comportamento de aranhas. Interciência, Rio de Janeiro, Brasil, pp. 67–87. 
Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A. & Huelsenbeck, J.P. (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61: 539–542. https://doi.org/10.1093/sysbio/sys029 
Roy, H.E., Steinkraus, D.C., Eilenberg, J., Hajek, A.E. & Pell, J.K. (2006) Bizarre interactions and endgames: entomopathogenic fungi and their arthropod hosts. Annual Review of Entomology 51: 331–357. https://doi.org/10.1146/annurev.ento.51.110104.150941 
Samson, R.A. & Evans, H.C. (1973) Notes on entomogenous fungi from Ghana: I. The genera Gibellula and Pseudogibellula. Acta Botanica Neerlandica 22: 522–528. https://doi.org/10.1111/j.1438-8677.1973.tb00873.x 
Samson, R.A. & Evans, H.C. (1992) New Species of Gibellula on Spiders (Araneida) from South America. Mycologia 84: 300–314. https://doi.org/10.1080/00275514.1992.12026143 
Samson, R.A., Evans, H.C. & Latge, J.-P. (2013) Atlas of Entomopathogenic Fungi. Springer Berlin / Heidelberg, Berlin, Heidelberg.
Shrestha, B., Kubátová, A., Tanaka, E., Oh, J., Yoon, D.-H., Sung, J.-M. & Sung, G.-H. (2019) Spider-pathogenic fungi within Hypocreales (Ascomycota): their current nomenclature, diversity, and distribution. Mycological Progress 18: 983–1003. https://doi.org/10.1007/s11557-019-01512-3
Souza, M.J.N. & Oliveira, V.P.V. (2008) Os enclaves úmidos e sub-úmidos do semi-árido do nordeste brasileiro (humid and sub-humid segments of the semi-arid area of the brazilian northeast). Mercator 5. 
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 
Tzean, S.S., Hsieh, L.S. & Wu, W.J. (1997) The genus Gibellula on spiders from Taiwan. Mycologia 89: 309–318. https://doi.org/10.2307/3761088 
Tzean, S.S., Hsieh, L.S. & Wu, W.J. (1998) Torrubiella dimorpha, a new species of spider parasite from Taiwan. Mycological Research 102: 1350–1354. https://doi.org/10.1017/S0953756298006467 
Ulloa, M. & Hanlin, R.T. (2000) Illustrated dictionary of mycology. American Phytopathological Society. 
Vega, F.E. & Blackwell, M. (2005) Insect-fungal associations: ecology and evolution. Oxford University Press, New York. 333 pp. 
Vega, F.E., Goettel, M.S., Blackwell, M., Chandler, D., Jackson, M.A., Keller, S., Koike, M., Maniania, N.K., Monzón, A., Ownley, B.H., Pell, J.K., Rangel, D.E.N. & Roy, H.E. (2009) Fungal entomopathogens: new insights on their ecology. Fungal Ecology 2: 149–159. https://doi.org/10.1016/j.funeco.2009.05.001 
Vieira, C., Japyassú, H.F., Santos, A.J. & Gonzaga, M.O. (2007) Teias e forrageamento. In: Gonzaga, M.O., Santos, A.J. & Japyassú, H.F. (eds.) Ecologia e comportamento de aranhas. Interciência, Rio de Janeiro, Brasil, pp. 45–65. 
Vilgalys, R. & Hester, M. (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238–4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990 
Vilgalys, R. & Sun, B.L. (1994) Ancient and recent patterns of geographic speciation in the oyster mushroom Pleurotus revealed by phylogenetic analysis of ribosomal DNA sequences. Proceedings of the National Academy of Sciences 91: 4599–4603. https://doi.org/10.1073/pnas.91.10.4599 
Wheeler, W.C., Coddington, J.A., Crowley, L.M., Dimitrov, D., Goloboff, P.A., Griswold, C.E., Hormiga, G., Prendini, L., Ramírez, M.J., Sierwald, P., Almeida-Silva, L., Alvarez-Padilla, F., Arnedo, M.A., Silva, L.R.B., Benjamin, S.P., Bond, J.E., Grismado, C.J., Hasan, E., Hedin, M., Izquierdo, M.A., Labarque, F.M., Ledford, J., Lopardo, L., Maddison, W.P., Miller, J.A., Piacentini, L.N., Platnick, N.I., Polotow, D., Silva-Dávila, D., Scharff, N., Szűts, T., Ubick, D., Vink, C.J., Wood, H.M. & Zhang, J. (2016) The spider tree of life: phylogeny of Araneae based on target-gene analyses from an extensive taxon sampling. Cladistics 33: 574–616. https://doi.org/10.1111/cla.12182 
White, T.J., Bruns, T., 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 Application. Elsevier, Academic Press, New York, pp. 315–322.