The first locality of Chalciporus rubinus ( Boletales , Basidiomycota ) in Poland

Chalciporus rubinus (W.G. Sm.) Singer, described in 1868 from England, was found in a city park in Wroclaw. This is the first record of the species from Poland. Macro- and micromorphological characters of the Polish specimens are described and illustrated. The delimitation of Ch. rubinus , the knowledge of its distribution, ecology and conservation status is summarised.


MaTerIaL aND MeTHODS
Material was collected on one locality.The description of macroscopic features is based on fresh material, on seven collections, comprising more than 40 basidiomata in all stages of development.The microcharacters of three recorded basidiomata (from three collections: nr 1, 2, 6) were observed and measured under a light microscope at magnification 1500× (basidiospores) and 800× (other features).For microscopic observations, dried pieces of basidiomata were placed in 5% NH 4 OH for about 5 minutes, then transferred to deionised water until they become pliable.Free-hand sections of the rehydrated pieces of basidiomata were examined in 5%NH 4 OH, Congo red and phloxine (in 1%NH 4 OH).amyloidity was tested with the Melzer's reagent.Morphological measurements were made and are presented according to the method presented by Breitenbach and Kränzlin (1991).The abbreviation Q is the ratio of basidiospore length to its width.Terminology of morphological and anatomical elements has been adopted mainly from Vellinga (1988).reported size of basidiospores, basidia (with sterigmata) and cystidia (cheilocystidia, pleurocystidia, caulocystidia), as well as dimensions of pileipellis hyphae were based on 31, 21, 31 and 31 measurements, respectively.Basidiospore measurements, Q coefficient and cystidia are presented as the mean, standard deviation, with the minimum and maximum dimensions in parentheses.Dimensions of basidia are given as the range of minimum and maximum dimensions.Drawings were made with the aid of a drawing tube under an oil-immersion objective.The voucher specimens of Ch. rubinus have been deposited in the Herbarium of the Museum of Natural History, Wrocław university in Wrocław, poland (WrSL).

reSuLTS
Chalciporus rubinus was found for the first time in poland on 12 th of June 2007 in the Szczytnicki park, in the south-eastern central part of Wrocław (Fig. 1).Five carpophores growing on the ground in the neighbourhood of Tilia and Quercus were observed at the time.During the following forays made on 13 th , 19 th and 29 th of June 2007 a few dozen of carpophores were discovered at the same locality and its nearest surroundings.On these occasions Ch. rubinus was found under Fagus, Fraxinus, Quercus, as well as Philadelphus.Later in august and September of the same year, the species was found at the same locality again.Further observations confirming the occurrence of Ch. rubinus at the investigated site were conducted in the period between July and September 2008.The location of the species has been carefully marked and will be monitored in future.Macroscopic and Microscopic characters.pileus 14-85 mm in diameter, hemispherical, subumbonate then plano-convex to even applanate and with reflexed margin; surface tomentose to finely fibrillose, dry and mat at first, lubricated and bright during wet weather, felted with age and even a little cracked during dry weather; brown, buff or alutaceous, usually keeping a pink tinge in about 5 mm from the margin.Tubes first adnate, then slightly decurrent, up to 6 mm long, first pink or locally light yellow, then crimson-red to pink, the colour not changing in contact with air.pores irregular, angular, up to 1 mm in diameter, concolorous with the tubes, progressively becoming red-rose from the margin to the centre on age, the colour not changing under pressure.Stipe 15-50×5-20 mm, solid, cylindrical with a tapering base or cylindrical, often slightly curved, smooth or somewhat ribbed by the decurrent tubes on the top, above concolorous with the pores, below red-rose to yellow, usually with chromeyellow basal mycelium.Context in pileus and stipe quite compact, slightly watery in pileus, fibrillous in stipe, in pileus whitish with reddish patches, in stipe whitish to yellow and warm yellow at the base.Smell indistinct or barely noticeable.Taste mild.Spore print pale brown.Basidiospores (4.1) 6.3 ± 0.7 (7.2) × (3.8) 4.3 ± 0.5 (6.4) μm, Q = (0.68) 1.47 ± 0.22 (1.76), ellipsoid to oblong, with a moderate hilar appendage, weakly pigmented, pale yellow (in 5%NH 4 OH), with a large guttula, inamyloid.μm, narrowly clavate, hyaline or containing small granules while immature, mostly with 4 sterigmata, without a basal clamp.Cystidia (cheilocystidia , pleurocystidia) (36.6) 47.1 ± 5.9 (58.2) × (4.9) 7.1 ± 1.6 (9.8) μm, narrowly  cylindrical to narrowly fusiform, potbellied, rarely narrowly clavate, erected or slightly curved, sometimes flexuose, dirty yellow (in 5%NH 4 OH), somewhat encrusted with crystalline bodies.Caulocystidia (10.1) 27.0 ± 12.1 (53.5) × (2.2) 4.1 ± 1.4 (6.9) μm, cylindrical to clavate, usually articulate, mostly with obtuse apex, dirty yellow (in 5% NH 4 OH), accompanied by some scattered caulobasidia.pileipellis: a trichoderm transiting into a cutis, made up of usually interwoven, septate, cylindrical and thin-walled hyphae without clamps and with rounded, usually slightly broader (narrowly clavate to clavate) terminal elements (5,1-12) μm wide, with scattered incrustations and yellowbrown intracellular pigment (Figs 2, 3).  of the species belonged to the Group II, Subtomentosi, of Fries (1836-1838), and he described and illustrated the fungus under the name Boletus rubinus (Smith 1868).With improved knowledge of microcharacters and their application as generic criteria, the species have been placed in Suillus Gray (Kuntze 1893(Kuntze -1898;;Singer 1965), Xerocomus Quél.(Skirgiełło 1960), Rubinoboletus pilát & Dermek (pilát, Dermek 1969) and Chalciporus Bataille (Moser 1983;allesio 1985;Singer 1986;Horak 2005;Muñoz 2005;Klofac, Krisai-Greilhuber 2006).The approach of pilát and Dermek (1969) was adopted by some authors of the most recent mycological papers (e.g., Šutara 2005;Knudsen, Taylor 2008;Šutara 2008).according to pilát and Dermek (1969), the position of Boletus rubinus W.G. Sm. in such genera as Xerocomus, Suillus and Chalciporus was isolated; the common feature of these genera is the fact that all their species have a uniform, elongate boletoid shape of spores.Therefore, the transfer of this species to an independent genus, Rubinoboletus, seemed to be a fairly acceptable solution (Šutara 2005).

DeSCrIpTION OF THe SpeCIMeNS
Nevertheless, the autonomous generic status of Rubinoboletus is still controversial.Šutara (2005) came to a conclusion that, with regards to the anatomical structure of the carpophores, Chalciporus is very similar to Rubinoboletus.He additionally found that it was very difficult to specify the boundary between the genus Chalciporus and Boletus subg.Xerocomus (Quél.)Maubl.He gave the spore print colour as the main but rather doubtful criterion to distinguish between european representatives of Chalciporus (cinnamon-brown or ferruginous-brown) and Xerocomus (brownish with more or less strong olive tinges), and separated Rubinoboletus on the basis of its short spores.
The smaller spore size was not regarded as an important feature at the generic level by Singer (1986).Degreef and De Kesel (2008) made a discovery of an interesting representative of Chalciporus in africa.They described the species under the name Chalciporus africanus J. Degreef & De Kesel and reported that it was similar to the temperate Ch. rubinus (W.G.Sm.) Singer, but differed in its larger and more elongated spores, unchanging context and prominent reddish pileus colour.as a consequence, Degreef and De Kesel (2008) treated Ch. africanus as taxon bridging the generic difference between Chalciporus and Rubinoboletus pilát & Dermek.In their opinion the close relationship between Ch. africanus and Ch.rubinus confirms Singer's (1986) opinion that Boletus rubinus W.G. Sm. is undoubtedly a good species of Chalciporus, making Rubinoboletus a synonym of Chalciporus.Degreef and De Kesel (2008) maintained that all globose-spored taxa subsequently combined in, or described under Rubinoboletus should be placed elsewhere.Moreover, they supported the Corner's (Corner 1972 after, Singer 1986) statement that "subglobose spores are to be expected in any alliance of elongated spores".Klofac and Krisai-Greilhuber (2006) took a similar approach; they proposed to include the genus Rubinoboletus as a sub-genus in Chalciporus.This point of view seems to be the most reasonable taxonomical concept and is also kept here.
Besides Ch. rubinus, two other species of the genus occur in europe: Ch. piperatus (Bull.: Fr.) Bataille (syn.Ch. hypochryseus (Šutara) Courtec)) and Ch.amarellus (Quél.)Bataille (syn.Ch. pseudorubinus (Thirring) pilát & Dermek; Ch. pierrhuguesii (Boud.)Bataille) (Klofac, Krisai-Greilhuber 2006).For identification, the basidiospores, the taste of flesh, the colour of tubes and pores are the most important features.Ch. rubinus differs from other species of the genus mainly in the smaller size and broad elliptical to ovoid-spherical shape of basidiospores (Q < 2).Other significant features are not peppery (or even bitter) context, red-pink (reddish-ochraceous in age) pileus, carmine-red tubes and pores, red or carmine-red stipe with chrome-yellow base and occurring under deciduous trees (Muñoz 2005).Mild taste is also a specific feature of Ch. amarellus, while the distinct peppery taste is a distinctive character of Ch. piperatus.Fruiting bodies of Ch. amarellus are characterized by cream to pale orange-brownish or yellow-brownish (and sometimes pinkish-cream along the margin) pilei, pale yellow to intense yellow stipes and pink or reddish-pink (ochre with age) tubes and pores.The species is associated with Abies, Picea or Pinus and has a tendency to occur at high altitudes in the mountains (Gminder 1994;Horak 2005;Muñoz 2005).Ch. piperatus, besides its distinct peppery taste, is characterized by dark red to rusty-brown pores, rusty-yellow to reddish-ochre pileus and concolorous, but normally lighter, bright yellow or chrome-yellow towards the base stipe (Horak 2005;Muñoz 2005).The species seems to be associated with coniferous (Pinus, Picea, Abies) and deciduous trees (Quercus, Fagus, Betula, Castanea), but its mycorrhizal status is regarded as doubtful (Högberg et al. 1996;Knudsen, Taylor 2008).Some authors suspected Ch. piperatus of an association with Amanita muscaria (L.: Fr.) Lam.(Spooner, roberts 2005).The relation of these species was confirmed by Veerkamp and arnolds (2008), although the mechanism still has to be explained.
Ch. rubinus is known hitherto only from europe.In the 1960s it was considered to be a very rare taxon, known only from england, former Czechoslovakia and Germany (Saxony) (Singer 1965).Nowadays the species is widespread but regionally usually treated as very rare (Muñoz 2005;arnolds, Veerkamp 2008).The present distribution area of Ch. rubinus extends mainly throughout the West to Central europe, but localities scattered in southern and northern parts of the continent are also known.It is known from england (Legon et al. 2005), Germany (Kreisel 1987;Kleine et al. 2004), the Czech republic (polčák 2003;Skála 2003;antonín et al. 2006), the Netherlands (Keizer 1995;Beenen et al. 2002), Belgium (Van de Kerckhove 2001; Van de Kerckhove, Walleyn 2006), Bulgaria (assyov, Denchev 2004), Slovakia (Lizoň 2001), austria, Hungary (allesio 1985;Muñoz 2005) as well as Italy (allesio 1985), Spain (rubio et al. 2006) andNorway (Bendiksen et al. 1999).recently some authors have drawn mycologists' attention to changes in the distribution of Ch. rubinus.antonín et al. (2006) has reported the species as an example of spreading taxon, gradually increasing its previous limited distribution range in the Czech republic.This tendency has also been confirmed by Kreisel (2006) in Germany.Nevertheless, earlier recognized as a rare species, Ch. rubinus was included in the red lists of the above and other european countries.In the Czech republic, Germany and Bulgaria it is treated as an endangered species (Benkert et al. 1992;Gyosheva et al. 2000;Lizoň 2001), in Great Britain, Norway and Slovakia it is regarded as vulnerable (Bendiksen et al. 1999;Lizoň 2001;Legon et al. 2005), while in the Netherlands as near threatened (arnolds, Veerkamp 2008).
For many years fungus forays have remained concerned with potentially rich habitats in order to record the widest range of species diversity.as a result, they focused on rural sites, leaving urban areas with only occasional recording.In order to find a new localities of Ch. rubinus it is necessary to investigate man-made landscape.anthropogenic habitats may usually appear worthless for fungi, but actually they present a variety of challenges for studies.