A First Lichenicolous Corticiumspecies (Corticiaceae, Corticiales), Described from Thamnoliain Switzerland
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A first lichenicolous Corticium species (Corticiaceae, Corticiales), described from Thamnolia in Switzerland Paul Diederich1, Erich Zimmermann2, Masoumeh Sikaroodi3, Masoomeh Ghobad-Nejhad4 & James D. Lawrey3 1 Musée national d’histoire naturelle, 25 rue Munster, L–2160 Luxembourg ([email protected]) 2 Scheunenberg 46, CH-3251 Wengi, Switzerland ([email protected]) 3 Department of Biology, George Mason University, Fairfax, VA 22030-4444, U.S.A. ([email protected]; [email protected]) 4 Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), P. O. Box 3353-5111, Tehran 3353136846, Iran ([email protected]) Diederich, P. & E. Zimmermann, M. Sikaroodi, M. Ghobad-Nejhad & J. D. Lawrey, 2018. A first lichenicolous Corticium species (Corticiaceae, Corticiales), described from Thamnolia in Switzerland. Bulletin de la Société des naturalistes luxembourgeois 120 : 49–56. Abstract. The new lichenicolous fungus, Corticium silviae Diederich, E.Zimm. & Lawrey is described from Switzerland, where it grows on Thamnolia. Phylogenetic results also suggest that Limonomyces should best be regarded as a synonym of Laetisaria, a genus that has a sister position to Marchandiomyces. The following new combinations are proposed:Laetisaria buckii (Diederich & Lawrey) Diederich, Lawrey & Ghobad-Nejhad (= Marchandiomyces buckii), L. culmigena (R.K.Webster & D.A.Reid) Diederich, Lawrey & Ghobad-Nejhad (= Exobasidiellum culmigenum), L. marsonii (Diederich & Lawrey) Diederich, Lawrey & Ghobad-Nejhad (= M. marsonii), L. nothofagicola (Diederich & Lawrey) Diederich, Lawrey & Ghobad-Nejhad (= M. nothofagicola), L. roseipellis (Stalpers & Loer.) Diederich, Lawrey & Ghobad-Nejhad (= Limonomyces roseipellis). 1. Introduction circumscription of the genera Laetisaria and Marchandiomyces should be revised. Most lichenicolous fungi belong to the asco- mycetes, while only five percent of them are basidiomycetes. These include species from 2. Material and Methods different orders and classes, the most diverse being the Tremellales, Cantharellales and 2.1. Morphological examination Corticiales (Diederich et al. 2018). A total of Type material of the new species is deposited four lichenicolous species were known from in G and in the private collection of P. Died- the Corticiaceae, belonging to Erythricium erich. External morphology was examined J.Erikss. & Hjortstam (= Marchandioba sidium Diederich & Schultheis), Laetisaria using a Leica MZ 7.5 dissecting microscope. Burds. and Marchandiomyces Diederich & Macroscopic photographs were taken using D.Hawksw. Two of these are known only a Canon 40D camera with a Canon MP-E 65 from the asexual bulbil-forming stage, one macro lens, StackShot (Cognisys) and Heli- only from the sexual stage, and one from con Focus (HeliconSoft) for increasing the both stages. depth of field. Microscopical structures were Here we report the discovery of a further studied using hand-cut sections stained with lichenicolous sexual species of Corticiaceae, a mixture of phloxine, Congo red and KOH discovered on Thamnolia in Switzerland. (5%). Microscopic photographs were pre- Phylogenetic analyses based on nuLSU and pared using a Leica DMLB microscope with ITS sequences suggest that the new species DIC, using a Leica EC3 camera and Heli- belongs to the genus Corticium and that the con Focus. Basidiospore measurements are Bull. Soc. Nat. luxemb. 120 (2018) 49 indicated as (min.–) X-σ – X+σ (–max.), fol- difficulty aligning ITS sequences available lowed by the number of measurements (n). in GenBank, so we trimmed our sequence to include only the nuLSU and used only 2.2. Molecular data published nuLSU sequences from Gen- Tissue scraped from the specimen was Bank in our analyses. The final data set washed in 70% ethanol and then genomic (all GenBank accession numbers included DNA was extracted using the Fast DNA in Fig. 1) contained 35 ingroup terminals, Spin Kit from MP Biomedicals (Santa Ana, and sequences from Gloeophyllales [Velu CA) according to the manufacturer’s pro- ticeps abietina (Pers.) Hjortstam & Tellería, tocol. About 10 ng of extracted DNA were Heliocybe sulcata (Berk.) Redhead & Ginns, subjected to a standard PCR in a 20 µL Gloeophyllum sepiarium (Wulfen) P.Karst., reaction volume using Taq Gold polymer- and G. abietinum (Bull.) P.Karst.] were used ase (Applied Biosystems, Foster City, CA), as outgroups. also according to manufacturer’s protocols, The newly generated nuLSU sequences with the objective of amplifying the ITS and were edited in Geneious v.8.1.6 (http:// nuclear large subunit (nuLSU) rDNA. The www.geneious.com/) and automatically products were purified with Ampure mag- aligned with MAFFT using the --auto netic beads (Agencourt Bioscience, Beverly, option (Katoh & Toh 2005). The align- MA) and the purified PCR products were ments were trimmed and subjected to used in standard sequencing reactions with analysis of ambiguously aligned regions BigDye Terminator Ready Reaction Mix using the GUIDANCE webserver (Penn et v3.1 (Applied Biosystems). The sequenc- al. 2010a,b); regions aligned with low confi- ing reactions were purified using Sephadex dence (below 0.93) were removed. The final G-50 (Sigma-Aldrich, St. Louis, MO), dried nuLSU data set had an alignment length of in a speedvac, denatured in HiDi Forma- 862 bases, 231 of which were variable. The mide (Applied Biosystems) and run on an GUIDANCE score for the nuLSU align- ABI3130-xl capillary sequencer (Applied ment was 0.9622. Maximum likelihood Biosystems). The data collected were ana- (ML) searches were done using RAxML lyzed using ABI Sequencing Analysis v5.4 7.2.6 (Stamatakis 2006; Stamatakis et software, and the sequences were then al. 2005) with non-parametric bootstrap- assembled together with the Sequencher ping of 1000 replicates under the universal version 5.0.1 (Gene Codes, Ann Arbor, MI) GTRGAMMA model. A Bayesian analysis software for manual corrections in base call- was also performed for the same data sets ing and to make contiguous alignments of using Markov chain Monte Carlo sam- overlapping fragments. The primers used pling (Larget & Simon 1999) in MrBayes were LR0R, LR3R, LR5, LR7, LR16, ITS4 3.1.2 (Huelsenbeck & Ronquist 2001). A and ITS5 (http://www.biology.duke.edu/ substitution model was selected in jMod- fungi/mycolab/primers.htm). elTest 0.1.1 (Posada 2008), which employs PhyML 3.0 (Guindon & Gascuel 2003) to 2.3. Phylogenetic analysis estimate the likelihood of the data under 24 In addition to our newly generated models of evolution using a fixed topology. sequence (GenBank accession: Corti The AICc values under each model were cium sp. ITS and nuLSU: MH520061), we compared and the model with the lowest included sequences used in earlier analyses AICc value (GTR+I+G) was selected. Two of the Corticiales (Sikaroodi et al. 2001; parallel analyses were then run in MrBayes DePriest et al. 2005; Lawrey et al. 2007, for 10,000,000 generations, with 4 chains 2008; Ghobad-Nejhad & Hallenberg 2011; each, sampling every 100 generations. Ghobad-Nejhad et al. 2010) and those Burn-in trees (initial 25%) were discarded from a broad range of taxa representing for each run and posterior probabilities recognized clades in the Corticium clade (PP) of the nuLSU matrix were determined (Langer 2002; Larson 2004; Binder et al. by calculating a majority-rule consensus 2005; Hibbett et al. 2007, 2014; Larson et tree generated from the post-burnin trees al. 2007; Diederich et al. 2011). We had by the MCMCMC runs using the sumt 50 Bull. Soc. Nat. luxemb. 120 (2018) Bull. Soc. Nat. luxemb. 120 (2018) luxemb. Soc.Bull. Nat. silviae of the placement species showing used of inthe analysis, RAxML phylogram Fig. 1. Best-scoring nuLSU 92 HQ168400_Laetisaria_lichenicola . Internal branches in boldface indicate posterior probabilities ≥0.95 and numbers are ML-BS values ≥70. values ML-BS are numbers ≥0.95 and probabilities posterior inboldface indicate branches . Internal 29 EU622839_Marchandiomyces_marsonii 65 EU622848_Limonomyces_culmigenus EU622844_Limonomyces_roseipellis 60 59 Laetisaria/Limonomyces AY293192_Laetisaria_fuciformis 100 DQ915474_Marchandiomyces_nothofagicola 73 DQ915472_Marchandiomyces_buckii 100 AY583331_Marchandiomyces_corallinus 100 HM046929_Marchandiomyces_quercinus Marchandiomyces AY583332_Marchandiomyces_lignicola 43 100 HM046919_Corticium_boreoroseum 77 HM046920_Corticium_boreoroseum 53 Corticium_silviae 97 EF537893_Corticium_roseum AY463401_Corticium_roseum Corticiaceae 77 100 HQ168397_Erythricium_aurantiacum 37 AY586655_Erythricium_laetum 100 AY542864_Lawreymyces_palicei Corticiales 30 AY542865_Lawreymyces_palicei 81 AY885164_Waitea_circinata 25 69 EU622842_Laetisaria_arvalis AF506709_Erythricium_salmonicolor 100 AF518617_Galzinia_incrustans 65 AF518666_Vuilleminia_comedens 77 HM046928_Vuilleminia_pseudocystidiata 51 HM046923_Vuilleminia_cystidiata AY586726_Vuilleminia_macrospora Vuilleminiaceae 71 44 DQ915478_Cytidia_salicina HM046930_Australovuilleminia_coccinea 62 KJ668413_Dendrocorticium_roseocarneum 100 90 AF393053_Dendrocorticium_roseocarneum 100 AJ406531_Dendrocorticium_polygonioides 97 HM046932_Punctulariopsis_subglobispora Punctulariaceae 42 HM046933_Punctulariopsis_obducens AF518642_Punctularia_strigosozonata 83 AJ583432_Gloeophyllum_sepiarium 96 AJ583431_Gloeophyllum_abietinum 100 Corticium AF518619_Heliocybe_sulcata Gloeophyllales EU118619_Veluticeps_abietina 51 0.03 substitutions/site option of MrBayes. RAxML and MrBayes 3.2. Taxonomy analyses were performed using the CIPRES Corticium