View metadata,Downloaded citation and from similar orbit.dtu.dk papers on:at core.ac.uk Dec 19, 2017 brought to you by CORE provided by Online Research Database In Technology Rasamsonia, a new genus comprising thermotolerant and thermophilic Talaromyces and Geosmithia species Houbraken, J.; Spierenburg, H.; Frisvad, Jens Christian Published in: Antonie van Leeuwenhoek Link to article, DOI: 10.1007/s10482-011-9647-1 Publication date: 2012 Document Version Publisher's PDF, also known as Version of record Link back to DTU Orbit Citation (APA): Houbraken, J., Spierenburg, H., & Frisvad, J. C. (2012). Rasamsonia, a new genus comprising thermotolerant and thermophilic Talaromyces and Geosmithia species. Antonie van Leeuwenhoek, 101(2), 403-421. 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Antonie van Leeuwenhoek (2012) 101:403–421 DOI 10.1007/s10482-011-9647-1 ORIGINAL PAPER Rasamsonia, a new genus comprising thermotolerant and thermophilic Talaromyces and Geosmithia species J. Houbraken • H. Spierenburg • J. C. Frisvad Received: 1 March 2011 / Accepted: 16 September 2011 / Published online: 2 October 2011 Ó The Author(s) 2011. This article is published with open access at Springerlink.com Abstract The phylogenetic relationship among covering. Species within the genus Rasamsonia were Geosmithia argillacea, Talaromyces emersonii, Ta- distinguished using a combination of phenotypic laromyces byssochlamydoides and other members of characters, extrolite patterns, ITS and partial calmod- the Trichocomaceae was studied using partial RPB2 ulin and b-tubulin sequences. Rasamsonia brevistipi- (RNA polymerase II gene, encoding the second largest tata sp. nov. is described and five new combinations protein subunit), Tsr1 (putative ribosome biogenesis are proposed. protein) and Cct8 (putative chaperonin complex component TCP-1) gene sequences. The results Keywords Emersonii Á Argillacea Á Taxonomy Á showed that these species form a distinct clade within Extrolites Á Phylogeny Á Trichocomaceae the Trichocomaceae and Trichocoma paradoxa is phylogenetically most closely related. Based on phenotypic and physiological characters and molecu- Introduction lar data, we propose Rasamsonia gen. nov. to accom- modate these species. This new genus is distinct from Thermophilia and thermotolerance have been vari- other genera of the Trichocomaceae in being thermo- ously defined (Apinis 1963; Cooney and Emerson tolerant or thermophilic and having conidiophores 1964; Craveri et al. 1964; Crisan 1973). The com- with distinctly rough walled stipes, olive-brown monly accepted definition is that of Cooney and conidia and ascomata, if present, with a scanty Emerson (1964), who defined thermophilic species as those which can grow at or above 50°C, but not below 20°C. The definition of thermotolerance is similar Electronic supplementary material The online version of except that the maximum growth temperature is at or this article (doi:10.1007/s10482-011-9647-1) contains supplementary material, which is available to authorized users. above 45°C, having a maximum near 50°C, and the minimum growth temperature is below 20°C. Ther- J. Houbraken (&) Á H. Spierenburg mophilic fungi have gained much attention in bio- Department of Applied and Industrial Mycology, technology the last decades, primarily because of their CBS-KNAW Fungal Biodiversity Centre, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands capacity to produce thermostable enzymes, which are e-mail: [email protected] stable at room temperature and can be purified by heat treatment (Maheshwari et al. 2000; Turner et al. 2007). J. C. Frisvad For example, the thermophilic species Talaromyces Department of Systems Biology, Technical University of Denmark, Building 221, Søltofts Plads, emersonii produces multiple b-glucosidase enzymes, 2800 Kongens Lyngby, Denmark a number of which are components of a potent 123 404 Antonie van Leeuwenhoek (2012) 101:403–421 cellulytic enzyme system and are used in commercial TCP-1) gene sequences. These data indicate that these applications (Collins et al. 2007; Coughlan et al. 1993; species are closely related and phenotypic and physio- McHale and Coughlan 1981; Murray et al. 2004). logical characters show that they represent a new genus. Thermophilic fungi are not restricted to a specific The taxonomy of this new genus is studied here using a taxonomic order and occur all over the fungal tree of combination of phenotypic characters, extrolite pat- life. Examples are Rhizomucor in the Mucorales, terns, ITS and partial calmodulin and b-tubulin Myceliophthora in Sordariales and Talaromyces, sequences. One new species is described and five new Thermomyces and Thermoascus in the Eurotiales combinations are proposed. (Mouchacca 1997). The majority of Talaromyces species are mesophiles but exceptions are species within sections Emersonii and Thermophila. Section Materials and methods Emersonii includes Talaromyces emersonii, T. bys- sochlamydoides, T. bacillisporus and T. leycettanus, Strains all of which grow well at 40°C. Talaromyces bacil- lisporus is thermotolerant, T. leycettanus is thermo- The strains used in the study on the phylogeny of the tolerant to thermophilic, and T. emersonii and Trichocomaceae are listed in Supplementary Table 1. T. byssochlamydoides are truly thermophilic (Stolk An overview of strains used for phenotypic analysis and Samson 1972). The sole member of Talaromyces and phylogeny is shown in Table 1. All cultures are section Thermophila, T. thermophilus, grows rapidly maintained in the culture collection of the CBS- at 50°C (Evans and Stolk 1971; Evans 1971; Stolk and KNAW Fungal Biodiversity Centre, Utrecht, The Samson 1972). The current classification of these Netherlands. thermophilic Talaromyces species is mainly based on phenotypic and physiological characters, such as their Morphological characterization ability to grow above 40°C, ascospore color, the structure of ascomatal covering and the formation of a Cultures for morphological observation were inocu- certain type of anamorph. Stolk and Samson (1972) lated in a three point position onto the following agar stated that the members of the section Emersonii have media: Czapek yeast extract agar (CYA), CYA anamorphs of either Paecilomyces (T. byssochlamydoi- supplemented with 5% NaCl (CYAS), yeast extract des and T. leycettanus)orPenicillium cylindrosporum sucrose agar (YES), creatine sucrose agar (CREA), series (T. emersonii and T. bacillisporus). Later, dichloran 18% glycerol agar (DG18), oatmeal agar Pitt (1979) transferred the species belonging to the (OA) and malt extract agar (Oxoid) (MEA). All media Penicillium cylindrosporum series to the genus were prepared as described by Samson et al. (2010). Geosmithia, based on various characters such as the Two sets were inoculated per strain: one set was formation of conidia from terminal pores instead of incubated at 25°C, the other at 37°C. Colony diameter on collula (necks), a character of Penicillium and and other macroscopical features were recorded after Paecilomyces. Within the genus Geosmithia, only 7 days of incubation. Microscopical features were G. argillacea is thermotolerant, and Stolk et al. (1969) studied by light microscopy (Olympus BH2 and Zeiss and Evans (1971) proposed a connection with mem- Axioskop 2 Plus) and microscopic mounts were made bers of Talaromyces sect. Emersonii. The phyloge- from MEA or OA. netic relationship of the thermophilic Talaromyces species within Talaromyces and the Trichocomaceae Molecular analysis is unknown. In this study, the phylogenetic relationship among Strains were grown on MEA for 7–14 days prior to Geosmithia argillacea, T. emersonii, T. byssochlamydoi- DNA extraction. Genomic DNA was extracted using des and other thermotolerant/thermophilic members of the UltracleanTM Microbial DNA isolation Kit the Trichocomaceae is studied using partial RPB2(RNA (MoBio, Solana Beach, U.S.A.) according to the polymerase II gene, encoding the second largest protein manufacturer’s instructions. For the phylogenetic subunit), Tsr1 (putative ribosome biogenesis protein) analysis of the Eurotialean taxa, parts of the RPB2, and Cct8 (putative chaperonin complex component Tsr1 and Cct8 genes were sequenced. The latter two 123 Antonie van Leeuwenhoek (2012) 101:403–421 Table 1 Rasamsonia isolates used in this study CBS no. Name Other collections Origin GenBank accession numbers (ITS/BenA/Cmd) CBS 101.69T R. argillacea (= Geosm. argillacea) DTO 97E4 = IMI Mine tip with a very high JF417491; JF417456; JF417501 156096 = IBT 31199 surface temperature; Staffordshire, UK CBS 102.69 R. argillacea (= Geosm. argillacea) DTO 97E5 = IBT 31200 Air; UK JF417490; JF417457; JF417502 CBS 907.70 R. argillacea (= Geosm. argillacea) DTO 97E7 Unknown source; Reading, JF417474; JF417458;