18th Congress of European Mycologists Bioblitz 2019 – Naturalists Contribute to the Knowledge of Mycobiota and Lichenobiota of Białowieża Primeval Forest

A total of 561 records of 233 species of fungi are reported from the Polish part of Białowieża Forest as a result of a short-term inventory that was conducted during the 18th Congress of European Mycologists (September 18–29, 2019). Four species new to Poland (Bryocentria brongniartii, Tremella coppinsii, T. hypocenomycis, and Zevadia peroccidentalis), and eight species new to Białowieża Primeval Forest (Hypomyces chrysostomus, Hypomyces rosellus, Lachnellula resinaria, Peniophora lycii, Phellinus viticola, Phlebia subochracea, Pronectria anisospora, and Typhula quisquiliaris) were recorded.


Introduction
Białowieża Primeval Forest, covering approximately 1,500 km 2 , is one of the best preserved old-growth mixed forests in Europe. It is located on the Poland-Belarus border in the North Podlasie Plain (Solon et al., 2018). The preservation of Białowieża Forest was possible because from the fifteenth to the eighteenth century, it was used as a royal hunting reserve. Despite various extent of human use through the ages, large parts of the forest remained relatively free of human impact until the twentieth century (Jaroszewicz et al., 2019;Samojlik et al., 2013;Szwagrzyk, 2016). Modern nature conservation started in 1921 when a "Reserve" forestry was created in Białowieża, encompassing 45.9 km 2 of old-growth forest. In 1932, it was transformed into The National Park in Białowieża and in 1947, after World War II, it was restored as Białowieża National Park (Okołów et al., 2009). Białowieża Primeval Forest has been a UNESCO World Heritage Site since 1979 (initially only Polish and Belarusian national parks, the whole Białowieża Forest since 2014). The Polish part of Białowieża Primeval Forest is additionally an EU Natura 2000 site, which includes an overlapping Special Protection Area and Special Area of Conservation. The main purpose of most of these designated sites is to protect the natural processes in the remaining part of the primeval forest from direct human interference (Bobiec, 2002).
In the Polish part of Białowieża Primeval Forest, a lime-oak-hornbeam forest Tilio-Carpinetum is the dominant type of potential natural vegetation. However, only approximately half of the area is currently covered by mixed deciduous forests. The remaining areas were changed to Norway spruce or Scots pine plantations as a result of twentieth century forest management. Nonetheless, many well-preserved, old growth deciduous stands, alder carrs Carici elongatae-Alnetum, pine bogs Vaccinio uliginosi-Pinetum, and other, less common forest types, are protected within the nature reserves (Jaroszewicz et al., 2019;Sokołowski, 2004). In numerous areas, a natural mosaic of habitats and vegetation types have been preserved. Moreover, forest microhabitats such as deadwood, animal carcasses, and tree cavities, are abundant because of relatively low human disturbance (Jaroszewicz et al., 2019).
There is a long-standing history of mycobiota research in Białowieża Primeval Forest. The first studies of mycobiota in this area date back to the nineteenth century (Błoński, 1889;Błoński et al., 1888). Extensive inventory and taxonomic works in Białowieża National Park were conducted by various researchers in the twentieth century. Domański published a series of 18 papers entitled "Wood inhabiting fungi in Białowieża virgin forest" in Acta Societatis Botanicorum Poloniae between 1965 to 1972 (as cited in Karasiński & Wołkowycki, 2015). The dematiaceous hyphomycetes were investigated by Alina Borowska and her students, but the lists of species are available only as manuscripts (Kostecka, 1989;Szulc, 1995). An extensive and methodical study of lichens, including a summary of previous knowledge, was published by Cieśliński and Tobolewski (1988). The largest inventory project on the fungal diversity of this region, called CRYPTO, was conducted between 1987 and 1991. The project focused on one forest section (number 256) of Białowieża National Park and recorded 1,380 species of micro-, macrofungi, and lichens in an area of 144 ha (Bujakiewicz et al., 1992;Faliński & Mułenko, 1992, 1995, 1997. Knowledge about the lichenobiota of Białowieża Forest was first compiled in a red list of threatened lichens in Białowieża Old-Growth Forest , and later in a checklist of lichens of Białowieża Primeval Forest, which included 450 species (Cieśliński, 2010). Recently, numerous new species of lichens and allied fungi have been noted (e.g., Matwiejuk, 2011;Łubek & Jaroszewicz, 2012;Łubek et al., 2018).
The Global Biodiversity Information Facility (GBIF; https://www.gbif.org/) is currently the most widely used database for all kinds of biodiversity studies but exhibits similar limitations. The database currently contains 19,389,938 fungal observations representing 154,183 species. However, there are only 2,481 fungal records from the Polish part of Białowieża Forest. Of these, most (543 records) came from the iNaturalist citizen science application (https://www.inaturalist.org/). Rapid biodiversity inventories, called bioblitz, are a means to teach people about natural sciences and species in the field, while also raising awareness about the need for nature conservation. Bioblitzes generate important biodiversity datasets, especially if many expert naturalists (amateur or professional) are involved (Parker et al., 2018). Such inventories, called expert bioblitzes by Parker et al. (2018), have a long tradition in the Polish mycological community. Short field trips and fieldwork organized by the Mycological Section of the Polish Botanical Society, the Polish Mycological Society, national parks, museums, and amateur naturalist groups (e.g., bio-forum.pl; https://www.bio-forum.pl/), are rich sources of specimens and biodiversity data (e.g., Kujawa et al., 2015Kujawa et al., , 2018. Such rapid fungal inventories can record several hundred species from various taxonomic groups. However, the number of species recorded is highly dependent on the weather conditions and the specialists involved (e.g., Gierczyk et al., 2013;Kujawa et al., 2018;Ruszkiewicz-Michalska et al., 2015). Mycological meetings, like international congresses, offer unique opportunities to involve a large group of expert mycologists in species diversity studies. The first of these inventories was performed during the IV Congress of European Mycologists (Warsaw) in 1966 and resulted in the first exhibition of fungi from Białowieża Forest. Beginning in 1993, the current tradition of the annual mushroom exhibition of Białowieża Primeval Forest has continued. Recently, Kujawa et al. (2018) reviewed and analyzed all historical records of nonlichenized fungi from this region, including the data gathered from these annual exhibitions. The records were based on macro-and micromorphological observations of the collected specimens. Combining these data, Kujawa et al. reported 1,144 macrofungal taxa, of which 85 taxa were new to Poland. Altogether, approximately 2,200 species of macrofungi are known from the Polish part of Białowieża Forest (Kujawa, 2020;. At the 25th Jubilee exhibition in September 2019, 159 species of fungi were displayed and an additional 51 species were recorded. Among these were three species new to Poland (Athelopsis subinconspicua, Hypomyces microspermus, and Ophiocordyceps variabilis) and seven species new to Białowieża Forest .
Here, we report the diversity of fungi (including lichens and lichenicolous organisms) of Białowieża Primeval Forest as a result of a joint collection and identification effort from all participants of the 18th Congress of European Mycologists (CEM) in September 2019. The opportunity was seized to gather the diverse expertise of more than 200 international mycologists to collect and identify fungi using macro-and microscopic morphology, along with sequence data that has been made available in the GenBank and GBIF databases.

Material and Methods
Fungi were collected from managed areas of Białowieża Primeval Forest (Podlaskie Voivodeship, Poland) between September 19 and September 27, 2019 by participants of the 18th CEM and University of Warsaw students. Forest divisions 424 and 362 were sampled between September 19 and September 21, 2019 and divisions 403, 450A, 469C, and 491B between September 23 and September 27, 2019 ( Figure 1). Sampled sites represented lime-oakhornbeam forest habitat with the exception of divisions 469C and 491B, where secondary mixed forest prevailed (Forest Data Bank, 2020;Matuszkiewicz, 2002). Additionally, observations from excursions to Białowieża National Park were added. Observations made by iNaturalist are available on the 18th CEM project website: https://www.inaturalist.org/projects/xviiicem. iNaturalist records were manually checked for redundant observations and filtered out. All localities include a reference to the forest district and division if recorded from the managed part of Białowieża Forest (Figure 1). The nomenclature was aligned using the GBIF species lookup tool (https://www.gbif.org/tools/species-lookup), and later corrected to follow Mycobank (https://mycobank.org/; Robert et al., 2013). If the current name from Mycobank is different from GBIF or one in used checklists, the latter are given as synonyms in brackets. For selected specimens, morphological identification was confirmed by sequencing of ITS rDNA. The sequences obtained in this study were deposited in the International Nucleotide Sequence Database Collaboration under the following GenBank accession numbers: MT229985-MT229993, MT240483, and MT240484. Voucher specimens were deposited in the Herbarium of the Faculty of Biology, University of Warsaw (WA) and the Herbarium of Swiss Federal Institute of Technology in Zurich (ZT Myc). The data on species occurrences were also deposited in GBIF (Siedlecki & Pawłowska, 2020). All species records we report here are also available in the supplementary Table S1.
Finally, the protective status of all species were annotated according to the regulation of the Polish Minister of the Environment (Minister of Environment, 2014), the "Red list of the macrofungi in Poland" (Wojewoda & Ławrynowicz, 2006), the IUCN Red List (The IUCN Red List of Threatened Species, 2020), the Checklist of Polish Larger Basidiomycetes (Wojewoda, 2003), A Preliminary Checklist of Micromycetes in Poland (Mułenko et al., 2008), the Checklist of Microfungi and Larger Ascomycetes of Białowieża Forest (Kozłowska et al., 2019), "The lichens of Poland. A fourth checklist" (Fałtynowicz & Kossowska, 2016), the "Red list of extinct and threatened lichens in Poland" , and a literature database of macrofungi in Poland (Kujawa, 2020).

Results
The eight (three plus five) day inventory yielded 101 collections and 68 observations representing 142 species. During the 11 days of the dedicated bioblitz project on iNaturalist, 750 observations representing 223 species were recorded, out of which 418 observations were of research grade. After removing redundant observations and doubtful identifications, 392 records representing 161 species were obtained from iNaturalist (only these data will be further included in the current paper). In total, 561 observations representing 233 species were recorded. ITS rDNA sequences were obtained for 11 specimens: Crucibulum crucibuliforme, Mycena inclinata, Nectria cinnabarina, Pholiota sp., Postia stiptica, Steccherinum fimbriatum, Stereum subtomentosum, Trichaptum abietinum, Tremella aurantia, Tyromyces chioneus, and Xylaria sp.
All of the species noted for the first time from the Białowieża region or even from Poland form small fruit bodies; therefore, they were probably overlooked during previous investigations. Many fungal species are hard to identify or inconspicuous, and are thus left unrecorded by nonspecialists. This often results in an expertdistribution record of some taxa rather than species distribution.   Döbbeler, f. dist. Białowieża, div. 424, 52.7084N, 23.7879E, 2019-09-20, leg. 18thCEM, det. BSI, ZT Myc 60832; Figure 2D-F. Notes: Bryocentria brongniartii is a representative of the Bionectriaceae family which is characterized by the formation of small, orange perithecia. It parasites on liverworts, such as Frullania (Döbbeler, 2004(Döbbeler, , 2010 and is known from the northern hemisphere. There are 26 occurrences from Austria, Germany, Great Britain, Ireland, Italy, Norway, Spain and the USA reported in GBIF (2020).

Supporting Material
The following supporting material is available for this article: • Table S1. Detailed table of all records with metadata.