Ex situ conservation of rare and endangered species in mushroom culture collections of Ukraine

Biodiversity loss, degradation of ecosystems and other global concerns emphasize urgent issues in nature conservation. Fungi as an important component of biodiversity need to be protected as much as other living organisms. In the article, two basic strategies in conservation are outlined and recent advances in fungal conservation, both in situ and ex situ, are considered. The key role of mushroom culture collections for ex situ fungal conservation is highlighted. Conservation of species diversity and genetic resources in culture collections provides the essential basis for biotechnological potential. The largest collections of fungi registered in the World Data Center for Microorganisms (WDCM), a unique global network for ex situ preservation of fungal and microbial diversity, are mentioned. Of the existing culture collections of fungi in Ukraine, the IBK and FCKU mushroom culture collections are characterized. A list of the species maintained in the IBK and FCKU collections, evaluated in the European countries against the IUCN criteria and categories and available at the European Council for Conservation of Fungi (ECCF) website as the preliminary European Red List of endangered macrofungi, is provided. A special attention paid in the IBK and FCKU collections to cultures of rare species of fungi, particularly of those listed in the Red Data Book of Ukraine, as well as the opportunity of the reintroduction into natural habitats are briefly discussed. Some examples of other rare species of fungi from these collections, potential candidates for national red-listing, are presented.


Гриби і грибоподібні організми Fungi and Fungi-like Organisms
Conservation of biological diversity is one of the key concerns of the present day.The Convention on Biological Diversity covers biodiversity at various levels from species to ecosystems and genetic resources.Fungi are known to be an important component of biodiversity at all these levels.Nonetheless, until the recent past fungi were rarely considered in conservation issues compared to plants or animals and were often overlooked in international biodiversity agreements.However, during the last two decades awareness of fungal conservation has noticeably raised.
The European Council for Conservation of Fungi (ECCF) founded in 1985 was the world's first organization to promote fungal conservation in Europe both on continental and national levels.Over about a 30-year period, the ECCF members have made considerable contribution to fungal conservation, advanced general awareness of endangered species of fungi and their habitats.As a conservation wing of European Mycological Association (EMA) since 2003, the ECCF enhanced mycological infrastructure within Europe through a network of experts dedicated to conservation of fungi.By 2010, mycologists in 33 European countries have conducted Red List assessments of fungi which resulted in 5500 species nationally red-listed (Dahlberg et al., 2010).A shortlist of about 1700 species relevant for Red List evaluation at the European level is currently available at the ECCF website (http://www.eccf.eu/activitiesen.ehtml).A great advantage of the ECCF activities was that applying the IUCN (International Union for Conservation of Nature) categories and criteria for Укр. бот. журн., 2018, 75(4) strains of fungi of various taxonomic and ecological groups.
Among culture collections of macrofungi in Europe, Culture Collection of Basidiomycetes (CCBAS) of the Institute of Microbiology, Academy of Sciences of the Czech Republic, holds 292 species of the orders Agaricales and Aphyllophorales; of two collections registered in the Russian Federation, the All-Russian Collection of Microorganisms (VKM) maintains 164 species of Agaricales, and the Basidiomycetes Culture Collection of the Komarov Botanical Institute, Russian Academy of Sciences (LE-BIN) -633 species of Agaricales and Aphyllophorales (Psurtseva, 2008a, b;Psurtseva, Ozerskaya, 2013;Lomberg et al., 2015).These collections preserve the gene pool of a wide taxonomic diversity of mushrooms in culture that can be used in fundamental and applied science, development of bioeconomy and education purposes (Belova et al., 2005;Psurtseva, 2008a, b;Ozerskaya, 2012).
In Ukraine, there are several pure culture collections of macrofungi varied by number of strains and species composition.The IBK Mushroom Culture Collection of the M.G.Kholodny Institute of Botany, National Academy of Sciences of Ukraine is the largest fungal culture collection in Ukraine.Founded in 1966, it holds currently over 1110 strains of 186 species belonging to 88 genera of fungi, Basidiomycota and Ascomycota (Bisko et al., 2016a, b).This unique collection maintains dicaryotic strains of mushrooms from various taxonomic and environmental groups of a wide geographical origin.In 2001, the Cabinet of Ministers of Ukraine defined the IBK Mushroom Culture Collection as the National Heritage of Ukraine (Decree № 1709, adopted on 19 December, 2001).The IBK Mushroom Culture Collection (Curator Dr.Sci.Bisko N.A.) is registered in the WDCM international database (http://www.wfcc.info/ccinfo/index.php/collection/by_id/1152).
Several mushroom culture collections were established at the universities in Ukraine.They are as follows: Collection of the Department of Plant Biology (FCKU) of the Educational and Scientific Centre "Institute of Biology and Medicine", Taras Shevchenko National University of Kyiv (Kyiv), Collections of the Oles Honchar Dnipropetrovsk National University (Dnipro), Tauria State Agrotechnological University (Melitopol), National Forestry and Wood-Technology University of Ukraine (Lviv), National Technical University of Ukraine "Igor Sikorsky Kyiv Polytechnic Institute" (Kyiv), Institute of Breedings and Genetics, the assessment whenever possible was encouraged.As a result, recently produced national Red Lists of fungi in many European countries at least partly meet the IUCN criteria.Moreover, red-listing efforts stimulate research initiatives on fungal distribution, population structure and dynamics, as well as provide necessary data for conservation decisions and actions (Dahlberg, Mueller, 2011).
Simultaneously with growing awareness of the importance of protecting fungi in natural habitats, or in situ conservation, there has been a greater understanding of the complementary strategy, ex situ conservation.Increasing loss of biodiversity, continued depletion of natural resources, on-going concerns over climate change and biosecurity issues highlight the value of culture collections of fungi and microorganisms as genetic resources.
The worldwide importance of culture collections was originally recognized at the First Conference on Culture Collections held in Ottawa, Canada, in 1962, attended by representatives from 28 countries.In 1970, the World Federation for Culture Collections (WFCC) was established; subsequently, the World Data Center for Microorganisms (WDCM) was founded as the WFCC data center.The WDCM has gradually developed into an extensive global network of microorganism resource centers.By September 2017, according to the WDCM database management system, Culture Collections Information Worldwide (CCINFO), 728 culture collections from 72 countries have been registered which maintain over 2.5 million of microorganisms cultures, including 806 408 cultures of fungi (http:// www.wfcc.info/ccinfo/).
The growing demands on culture collections for authenticated, reliable biomaterial promote higher quality standards of scientific services provided for the development of biotechnology.Recently, the Organisation for Economic Co-operation and Development (OECD) has recognized the global value of culture collections as a significant element in the development of a knowledge-based bioeconomy (www.oecd.org).Thus conservation of genetic resources and species diversity provides the essential basis for biotechnological potential.
Three species of the genus Sparassis Fr. (S. crispa, S. laminosa, and S. nemecii) known in Ukraine are held in culture in the IBK and FCKU collections (Berezovska et al., 2012;Mykchaylova, 2017).Only one of them, S. crispa, is currently listed in the Red Data Book of Ukraine, although S. laminosa and S. nemecii are also rarely recorded in our country and need to be protected.Sparassis nemecii is regarded as a species relevant for Red List assessment in some European and Asian countries.Recently, a new locality of this fungus in Ukraine was reported from Hutsulshchyna National Natural Park (Heluta et al., 2016).
There are four strains of Pleurotus nebrodensis (Inzenga) Quel. of various geographical origin in the IBK collection.Until 2014, this species was the only representative of the Fungi Kingdom in the IUCN Red List of Threatened Species (http://www.iucnredlist.org).The occurrence range of this mushroom was estimated as less than 100 square km; in addition, its population was very fragmented and a continuous decrease in localities with mature individuals was observed.Thus, it was evaluated globally in the IUCN Red List as a Critically Endangered (CR) species (Heluta, Hayova, 2014).
There are three strains of Fomitopsis officinalis in the IBK collection.This species is listed in the Red Data Book of Ukraine as Regionally Extinct (RE) in our country.The fungus used to be found in the Carpathian and Western Ukrainian forests; however, for over 50 years it has not been recorded in Ukraine (Mykchaylova, 2016).Fomitopsis officinalis is considered as a species threatened with extinction in North America, 12 countries in Europe and the Asian part of the Russian Federation (Siberia, Far East).Only nine strains of this species are represented in the WFCC database.(http://gcm.wfcc.info/speciesPage.jsp?strain_ name=Fomitopsis%20officinalis).Five of them are stored in the CBS collection (the Netherlands), threein LE-BIN collection (Russian Federation) and one in MUT collection (Italy).
Apart from the species currently listed in the Red Data Book of Ukraine, other rare species of fungi can be regarded as candidates for red-listing.Some of them are continuously maintained in the mushroom National Academy of Agricultural Sciences of Ukraine (Odesa), Donetsk National University (Vinnytsia), etc.
The Culture Collection of Fungi (FCKU) of the Department of Plant Biology of the Educational and Scientific Centre "Institute of Biology and Medicine", Taras Shevchenko National University of Kyiv was founded in 2008 and at present holds 55 species of macrofungi of Basidiomycota and Ascomycota (Sukhomlyn, 2010).The FCKU Collection (Curator Prof., Dr.Sci.M.M. Sukhomlyn) is registered in the WDCM database (http://www.wfcc.info/ccinfo/collection/by_id/1000).One of the objectives of the FCKU Collection is preservation of rare species of macrofungi of Ukraine.It is the only fungal collection in Ukraine containing a pure culture of Laetisaria fuciformis, a potentially dangerous invasive species first recorded in Ukraine in 2006 (Akulov, 2010).
As mentioned above, ex situ method is a prospective approach for conservation of fungi.Its major purpose is to preserve the gene pool of macrofungi in pure culture.Hence, the fungal collections maintain taxonomic diversity of fungi, with an emphasis on rare and endangered species (Psurtseva, 2008a, b;Lomberg et al., 2015;Petrichuk et al., 2014a;Bisko et al., 2016b).In the IBK and FCKU collections, a special attention is also paid to cultures of rare species of fungi, particularly of those listed in the Red Data Book of Ukraine (2009).
For assessing the risk of species extinction, the criteria and categories proposed by the International Union for Conservation of Nature (IUCN) are commonly used; although originally designed for global level, they can be applied for fungal evaluation at the national level (Dahlberg, Mueller, 2011;Hayova, 2014).The existing Ukrainian Red Data Book categories should be consequently harmonized with international ones; the following IUCN categories are recommended to apply in the Red Data Book of Ukraine: Regionally Extinct (RE), Critically Endangered (CR), Endangered (EN), Vulnerable (VU), Near Threatened (NT), Least Concern (LC), and Data Deficient (DD) (Mosyakin et al., 2014).
The IBK and FCKU mushroom culture collections altogether support 124 species evaluated by selected European countries against the IUCN criteria and categories and other nationally accepted categories (ECCF, http://www.eccf.eu/),as well as those listed in the Red Data Book of Ukraine.A list of these species is provided below (Table ).
Culture collections provide an opportunity to reintroduce rare or declining species of fungi into natural habitats.For these purposes, preliminary studies on cultural properties of the species, including growth of vegetative mycelium and development of fruit bodies, are required to enable successful reintroduction.The research on reintroduction techniques has been conducted in Hutsulshchyna National Nature Park in Ukraine (Petrichuk et al., 2014a, b).
Through cooperation with Hutsulshchyna National Nature Park for three years, new strains of the rare and newly recorded species of macrofungi in Ukraine were added to the IBK and FCKU collections (Bisko et al., 2017).
Besides rare species described above, a large number of strains of edible fungi are sustained in the IBK and FCKU collections, such as Agaricus bisporus, Pleurotus ostreatus, Lentinula edodes, Flammulina velutipes, etc. (Lomberg et al., 2015;Bisko et al., 2016).These species are widely cultivated throughout the world.Most of them, except for L. edodes, grow in the wild in Ukraine and apparently do not require additional protection measures.However, they are considered as threatened or rare species in some European countries (Table ).
Thus, culture collections of macrofungi, containing numerous strains of mushrooms from various taxonomic and environmental groups of a broad geographical origin, are widely used in fundamental and applied science.The strains held in culture collections are of crucial importance in current taxonomic or phylogenetic studies as well as provide the essential basis for biotechnological potential.The research data on morphological and cultural characteristics of the strains contribute to reliable continuous preservation of the strains under cultural conditions for the purposes of ex-situ conservation, research, and sustainable use of genetic resources.

Species from the IBK and FCKU culture collections listed in the Red Data Book of Ukraine and preliminary European Red List of endangered macrofungi
collections.For example, the IBK and FCKU collections contain cultures of six species of the genus Hericium (Table).Several species of this genus (H.alpestre, H. clathroides, H. coralloides, and H. erinaceus) were recognized as threatened and therefore are red-listed in many European countries (http://www.eccf.eu/activities-en.ehtml).In our country, all four species are very rarely reported from national nature parks and nature reserves; however, only one of them, H. coralloides, is listed in the current edition of the Red Data Book of Ukraine (Petrichuk Тable.culture