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A Native of Dark Biofinishes on Oil Treated Wood Aureobasidium melanogenum Citation for published version (APA): van Nieuwenhuijzen, E. J., Houbraken, J. A. M. P., Meijer, M., Adan, O. C. G., & Samson, R. A. (2016). Aureobasidium melanogenum: a native of dark biofinishes on oil treated wood. Antonie van Leeuwenhoek, 109(5), 661-683. https://doi.org/10.1007/s10482-016-0668-7 DOI: 10.1007/s10482-016-0668-7 Document status and date: Published: 01/05/2016 Document Version: Publisher’s PDF, also known as Version of Record (includes final page, issue and volume numbers) Please check the document version of this publication: • A submitted manuscript is the version of the article upon submission and before peer-review. There can be important differences between the submitted version and the official published version of record. People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website. • The final author version and the galley proof are versions of the publication after peer review. • The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publication General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal. If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license above, please follow below link for the End User Agreement: www.tue.nl/taverne Take down policy If you believe that this document breaches copyright please contact us at: [email protected] providing details and we will investigate your claim. Download date: 26. Sep. 2021 Antonie van Leeuwenhoek (2016) 109:661–683 DOI 10.1007/s10482-016-0668-7 ORIGINAL PAPER Aureobasidium melanogenum: a native of dark biofinishes on oil treated wood Elke J. van Nieuwenhuijzen . Jos A. M. P. Houbraken . Martin Meijer . Olaf C. G. Adan . Robert A. Samson Received: 7 December 2015 / Accepted: 17 February 2016 / Published online: 27 February 2016 Ó The Author(s) 2016. This article is published with open access at Springerlink.com Abstract The genus Aureobasidium, which is presence of a biofinish and the type of substrate. A. known as a wood staining mould, has been detected melanogenum was detected on wood samples exposed on oil treated woods in the specific stain formation in the Netherlands, Cameroon, South Africa, Australia called biofinish. This biofinish is used to develop a and Norway. ITS-specific PCR amplification, cloning new protective, self-healing and decorative biotreat- and sequencing of DNA extracted from biofinish ment for wood. In order to understand and control samples confirmed results of the culturing based biofinish formation on oil treated wood, the occur- method: A. melanogenum is predominant within the rence of different Aureobasidium species on various Aureobasidium population of biofinishes on pine wood surfaces was studied. Phenotypic variability sapwood treated with raw linseed oil and the outdoor within Aureobasidium strains presented limitations of placement in the Netherlands. morphological identification of Aureobasidium spe- cies. PCR amplification and Sanger sequencing of ITS Keywords Aureobasidium pullulans Á Linseed oil Á and RPB2 were used to identify the culturable Mould staining Á Pine Á Sustainable Á Wood protection Aureobasidium species composition in mould stained wood surfaces with and without a biofinish. The analysed isolates showed that several Aureobasidium species were present and that Aureobasidium melano- Introduction genum was predominantly detected, regardless of the Aureobasidium are wood staining fungi, in particular on wood situated outdoors above the ground (Dickin- Electronic supplementary material The online version of son 1972; Dix and Webster 1995; Bardage 1998; this article (doi:10.1007/s10482-016-0668-7) contains supple- mentary material, which is available to authorized users. Schmidt 2006; Gobakken and Westin 2008). The interest in Aureobasidium has recently increased, E. J. van Nieuwenhuijzen (&) Á J. A. M. P. Houbraken Á because of its role in the formation of biofinishes on M. Meijer Á R. A. Samson wood (Sailer et al. 2010; van Nieuwenhuijzen et al. CBS KNAW Fungal Biodiversity Centre, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands 2013; van Nieuwenhuijzen et al. 2015; Filippovych e-mail: [email protected] et al. 2015). The term biofinish was introduced for a uniform dark mould covering which emerged outdoors O. C. G. Adan on oil treated wood (van Nieuwenhuijzen et al. 2015). Section Transport in Permeable Media, Department of Applied Physics, University of Technology Eindhoven, Although the protection mechanism and durability of Den Dolech 2, 5600 MB Eindhoven, The Netherlands this biofinish is still under investigation, biofinished 123 662 Antonie van Leeuwenhoek (2016) 109:661–683 wood is considered to be an appealing biocide-free GCPSR multigene phylogenies are required. Next to construction material that has the advantage of also the large subunit and the internal transcribed spacer having self-healing properties. regions (incl. 5.8S rDNA) (ITS) more variable genes Until now it is unknown which Aureobasidium such as translation elongation factor 1a, b-tubulin and species participates in the biofinish formation and RNA polymerase II- second largest subunit (RPB2) whether a biofinish is composed of more than a single have been applied or recommended for phylogenetic species. Although Aureobasidium has been isolated analysis of Aureobasidium species (Zalar et al. 2008; from many organic and inorganic substrates and Manitchotpisit et al. 2009; Peterson et al. 2013; geographical locations (Zalar et al. 2008; Slepecky Gostincˇar et al. 2014). A phylogeny, including all and Starmer 2009; Gaur et al. 2010), the ubiquity of described genera and species within the Aureobasidi- the specific species is unknown. The geographical aceae, is not yet available. location, the combination of wood species and the oil The ITS locus is assigned as the primary barcode treatment may all have an impact on the species for fungal species (Schoch et al. 2012). A large composition of the Aureobasidium population in number of ITS barcode sequences of Aureobasidium biofinishes. Species-specific behaviour, such as phe- species is available in public databases, which makes notype and physiology (Samson et al. 2010; Houbra- this DNA region a suitable marker to identify Aure- ken 2013), should be included in future research in obasidium (Manitchotpisit et al. 2009). To date, no order to understand and control dark mould growth on second fungal barcode has been determined for a oil treated wood. Therefore insight in the species reliable Aureobasidium identification on species level. composition of the biofinish is highly relevant. The aim of this study was to explore the Aureoba- The ascomycete genus Aureobasidium is a member sidium species composition of biofinishes on wood. of the family Aureobasidiaceae within the class of the Culturable Aureobasidium isolates, retrieved from Dothideomycetes (Thambugala et al. 2014; substrates with and without biofinishes, were identi- Wijayawardene et al. 2014). Kabatiella is closely fied. The wood species, oil treatments and exposure related to Aureobasidium based on morphology and sites were related to the culturable species composi- phylogeny (Zalar et al. 2008; Bills et al. 2012; Crous tion. Also direct extraction of biofinish DNA, followed et al. 2011; Peterson et al. 2013; Thambugala et al. by ITS amplification, cloning and sequencing were 2014) and some of these Kabatiella species may used to determine the species compositions of belong to Aureobasidium (Peterson et al. 2013; biofinishes. Thambugala et al. 2014). In addition future studies may result in the transfer of the species Selenophoma mahoniae and Columnosphaeria (Discosphaerina) Materials and methods fagi into Aureobasidium (Yurlova et al. 1999; Peterson et al. 2013; Thambugala et al. 2014). A well-known Substrates and outdoor exposure Aureobasidium species is Aureobasidium pullulans (Zalar et al. 2008; Gostincˇar et al. 2014). The total Nine sample sets were analysed in this study. Each set number of classified Aureobasidium species currently contained oil treated wood samples. Untreated pine varies per database, for example 38 in MycoBank and 13 in GenBank (October 2015). Before DNA sequencing was applied in fungal (oil absent) taxonomy, the species classification system was wood mainly based on physiologic and phenotypic charac- oil impregnaon teristics. In the case of Aureobasidium, colony blocks biofinish outdoor detecon pigmentation was used as a species-specific pheno- exposure & typic characteristic (Zalar et al. 2008; Peterson et al. moisture content sampling 2013). Nowadays, also the genealogical concordance glass sheets phylogenetic species recognition (GCPSR) concept is (oil & wood absent) commonly applied for species delimitation (Taylor Fig. 1 Flowchart illustrating the experimental setup of this et al. 2000). For species delimitation
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