Boletellus projectellus - An alien mycorrhizal bolete new to Europe

Abstract

Boletellus projectellus (Murill) Singer is recorded for the first time from Europe, based on collections made in Lithuania, Curonian Spit. Morphological fea¬tures and analysis of ITS sequences supporting the identity of the species are pro¬vided. Ecology, distribution and status of B. projectellus in Lithuania and in Europe are discussed.

Full text

203
Boletellus projectellus
– an alien mycorrhizal bolete
new to Europe
J. Motieju¯naite˙, J. Kasparavicˇius & A. Kacˇergius
1 Institute of Botany, Nature Research Centre, Žaliu˛ju˛ ežeru˛ 49, Vilnius,
LT-08406, Lithuania.
E-mails: jurga.motiejunaite@botanika.lt, jonas.kasparavicius@botanika.lt, audrius.
kacergius@botanika.lt
Motieju¯ naite˙ J., Kasparavicˇius J. & Kacˇergius A. (2011) Boletellus projectellus
– an alien mycorrhizal bolete new to Europe – Sydowia 63 (2): 203–213.
Boletellus projectellus (Murill) Singer is recorded for the rst time from Eu-
rope, based on collections made in Lithuania, Curonian Spit. Morphological fea-
tures and analysis of ITS sequences supporting the identity of the species are pro-
vided. Ecology, distribution and status of B. projectellus in Lithuania and in Eu-
rope are discussed.
Keywords: fungi, Basidiomycota, Boletaceae, phylogeography, invasions.
Within the past decades an unusual boletoid mushroom started to
attract mushroom-hunters’ attention at the northern part of the Curo-
nian Spit (Baltic Sea coast, Lithuania) and was dubbed a “red bolete”
by local people. During the last decade it has become common and
abundant in this area and increasing number of the specimens started
to arrive at the BILAS herbarium. Examination of the macro- and mi-
croscopical characters has shown that the fungus did not belong to any
boletoid species known in Lithuania or Europe and subsequently it
was identied as Boletellus projectellus (Murill) Singer, a species
known from eastern part of North America and SE Asia (Taiwan)
(Murrill 1938, Coker & Beers 1943, Singer 1945, Chen et al. 1998).
The purpose of this paper is to discuss morphological and ana-
tomical characteristics of B. projectellus in Lithuania, its ecology and
status as well as sequence data of the studied specimens.
Materials and Methods
Sampling site
All examined specimens were collected during the fall of 2007-
2009 in the northern part of the Curonian Spit, in Smiltyne˙ forest dis-
trict of Curonian Spit National Park. Further eld observations were
made in September and rst decade of October in 2010, in Smiltyne˙
and Juodkrante˙ forest districts.
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

204
The Curonian Spit is characterized by maritime climate; it is
strongly inuenced by the Baltic Sea. The climate is milder compared
to most continental parts of the country; average fall and winter tem-
peratures are by 3.0 °C to 3.5 °C higher than in the continental part,
thaws are common in winter and the number of sunny days is the high-
est in Lithuania. Air humidity reaches 82 % in winter and 76 % in
summer (Gudelis 1998). A large part (70 %) of the land is covered by
forests, more than half of them are planted. Conifers prevail (80 %)
with 53 % local Scots pine (Pinus sylvestris L.) and 27 % introduced
Mountain pine (Pinus mugo L.). Other introduced pines, like P. bank-
siana Lamb., P. nigra J.F. Arnold, P. strobus L. and others are found in
small groups or as solitary trees. The Curonian Spit is situated in the
migration route of the White and Baltic Seas, therefore millions of
birds pass it every year (Kuršiu˛ nerijos nacionalinio parko direkcija:
http://nerija.am.lt/VI/index.php#r/195). The northern part of the Spit
is also inuenced by sea transport: only several kilometres of the Curo-
nian Lagoon separate it from Klaipe˙da, one of the largest seaports in
the region.
Morphological and anatomical characteristics
Morphological features were observed and measurements were
taken from fresh basidiomata. Macrophotographs were taken by a Fuji
FinePix S5000 camera. Hymenium fragments taken from dried basidi-
omata were rehydrated and anatomical features measured in 3 % KOH
using a Zeiss Jenaval microscope (oil immersion objective 100 x); mi-
crophotographs were taken by a Nikon Coolpix 995 camera. Measure-
ments of pleurocystidia, basidia, spores and their length/width ratios
(Q) are given as follows: (minimum) mean ± standard deviation (maxi-
mum); values were rounded to one decimal place. Colours were subjec-Colours were subjec-
tively described and recorded following Kornerup & Wanscher (1978).
Voucher specimens are deposited in the mycological collection of BI-
LAS herbarium.
DNA extraction, amplication, sequencing and analysis
Genomic DNA was extracted from two frozen fresh fruiting bod-
ies (collected in 2009, part of the collections BILAS 48765 and 48766)
with NucleoSpin® Plant II Kit (Macherey–Nagel GmbH &Co. KG,
Germany) according to manufacturer’s instruction using approxi-
mately 100 mg wet weight of fruiting bodies. The internal transcribed
spacers 1 and 2 of rDNA, including the 5.8S rDNA, were amplied in
25 µl reactions on TProfessional 96 Gradient Thermocycler (Biometra
GmbH, Germany) in the following mixture: ~ 25 ng of template, 0.25
units of Taq polymerase (Fermentas UAB, Lithuania), 2.5 µl 10× PCR
buffer with KCl and MgCl2, 0.2 mM of each dNTP, 10 µM of primers
ITS5 and ITS4 (White et al. 1990). PCR conditions: 5 min at 95 °C as
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

205
initial denaturation, followed 35 cycles of 30 s at 94 °C, 30 s at 55 °C,
and 45 s at 72 °C, with nal extension of 10 min at 72 °C. Amplicons
were visualised under UV light in 1.5 % agarose gels stained with At-
lasSight DNA Stain (Bioatlas, Estonia). The PCR products were puri-
ed according to the Protocol for PCR Product Clean-up with Exonu-
clease I and FastAP™ Thermosensitive Alkaline Phosphatase (Fer-
mentas UAB, Lithuania). Puried PCR products were sequenced by
Macrogen (Macrogen Inc., Seoul, Korea) on an ABI 3730XL DNA se-
quencer. PCR products were successfully amplied with the estimated
size (692 characters). Four different PCR products from each specimen
with 2 repeats for each were sequenced from both ends (5’ and 3’) to
conrm the sequence. The rDNA homology searches (BLAST) were
performed through the internet at the National Center for Biotechnol-
ogy Information (National Institutes of Health, Bethesda, USA). Se-
quences were aligned by using Clustal W, and the phylogenetic tree of
the closely related sequences was constructed using the Lasergene
software package (DNASTAR, Inc., Madison, USA) by Neighbour-
Joining (NJ). Sequences derived in this study were deposited in the
NCBI GenBank database. The accession numbers and data are pro-
vided in Table 1.
Results and Discussion
Morphological and anatomical description
Boletellus projectellus
(Murill) Singer, Farlowia, 2: 129. 1945. – Figs.
1–5.
P il e u s 3 cm to 14 cm broad, convex, almost plane when older,
margin free, projecting and hanging straight down (Figs. 1–3), surface
dry, smooth, subtomentose, not cracked, colour brownish orange (6C4)
when young, becoming brownish red (8C8–9C8) with age. C o n t e xt
thick, up to 1.5 cm above the stipe, pale brownish-greyish orange (5B3)
turning slightly darker and ushed vinaceous near the cuticle when
cut. T ub e s pale yellow (3A3) when young, later olivaceous (3C8), 1.2
cm to 2 cm long, depressed around the stipe, not staining when cut or
bruised. P o r e s 1–2 mm broad, circular, yellow (3A4–3A5) when
young, nally pale olive (3B7–3C8). S ti p e 3 cm to 12 cm long, 1 cm to
3 cm thick, mostly tapering upward, with a coarse reticulum from top
to the bottom, base covered with white mycelium (Figs 2–3); surface
yellowish (4A4) when young, later becoming brownish yellow (5B5) to
reddish (7C7). F l e s h whitish to yellowish (3A2–3A3), buff (5B4) at
the base, slightly darkening on exposure. Odour not distinctive, taste
slightly acidic. S po r e p r i nt olivaceous brown (4D5). S p o r e s
(27.3) 30.1 ± 2.3 (36.4) x (7.8) 10.8 ± 1.2 (13.3) µm, Q = (2.1) 2.8 ± 0.3 (3.7)
(n = 51), ovate to fusiform, with a prominent suprahilar depression,
smooth, lacking a distinct apical pore, yellow in KOH (Fig. 4). B a-
s id i a (36.4) 41 ± 3.9 (44.2) x (12.7) 14.9 ± 1.1 (17.9) µm (n = 28), cla-
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

206
Tab. 1. – Names, accession numbers and reference details of the isolates studied
Species GenBank
accession No.
Country of origin, reference
Aureoboletus thibetanus
Boletus aereus
Boletus betulicola
Boletus betulicola betulicola
Boletus e�ulis e�ulis
Boletus e�ulis e�ulis
Boletus e�ulis e�ulis
Boletus e�ulis e�ulis
Boletus e�ulis e�ulis
Boletus e�ulis e�ulis
Boletus e�ulis e�ulis var. albus
Boletus e�ulis e�ulis var. aurantioruber
Boletus e�ulis var. clavipes
Boletus e�ulis var. pusteriensis
Boletus fechtneri
Boletus pinophilus
Boletus quercicolacicola
Boletus quercicola quercicola
Boletus reticulatus reticulatus
Morphotaxon A
Morphotaxon B
Morphotaxon C
Boletellus elatus
Boletellus elatus elatus
Boletellus emo�ensisemo�ensis
Boletellus mirabilis mirabilis
Boletellus mirabilis mirabilis
Boletellus mirabilis mirabilis
Boletellus mirabilis mirabilis
Boletellus longicolis longicolis
Boletellus longicolis longicolis
Boletellus obscurococcineus obscurococcineus
Boletellus projectellus projectellus
Boletellus projectellus projectellus
Boletellus russeli russeli
Boletellus shichianus shichianus
Unculture� Boletellus
Boletinellus exiquus
Boletinellus exiquus
Boletinellus merulioi�esoi�es
Boletinellus rompelii rompelii
Leccinum crocipo�ium
Leccinum crocipo�ium
Retiboletus nigerrimus
Suillus americanus
DQ200917
EU417845
EU417874
EU417846
EU417857
EU417852
EU417873
EU417853
EU417856
EU417870
EU417862
EU231978231978
EU231985
AY680985
EU417872
EU417865
EU417869
EU417867
EU417871
EU417850
EU417860
EU417863
AB509885
AB531456
AB509761
EU597076
AF335451
DQ367893
DQ384578
AB509647
AB509927
AB509989
AY789082
JN996470 –
JN996473
EU569284
DQ200921
FJ196901
AJ419183
AJ419145
DQ200922
AJ419192
AF454589
AF454590
AB509860
AF166502
China, Matheny et al. 2007
Netherlands, Beugelsdijk et al. 2008
Netherlands, Beugelsdijk et al. 2008
Finland, Beugelsdijk et al. 2008
Netherlands, Beugelsdijk et al. 2008
Finland, Beugelsdijk et al. 2008
Sweden, Beugelsdijk et al. 2008
Sweden, Beugelsdijk et al. 2008
Netherlands, Beugelsdijk et al. 2008
Netherlands, Beugelsdijk et al. 2008
Belgium, Beugelsdijk et al. 2008
USA, Dentinger & McLaughlin 2007
USA, Dentinger & McLaughlin 2007
Italy, Leonardi et al. 2005
Netherlands, Beugelsdijk et al. 2008
Sweden, Beugelsdijk et al. 2008
Austria, Beugelsdijk et al. 2008
Austria, Beugelsdijk et al. 2008
Netherlands, Beugelsdijk et al. 2008
Netherlands, Beugelsdijk et al. 2008
Netherlands, Beugelsdijk et al. 2008
Netherlands, Beugelsdijk et al. 2008
Japan, Sato et al. 2009
Japan, Sato et al. 2009
Japan, Sato et al. 2009
Canada, Jones et al. 2008
Canada, Berbee et al. 2001
Canada, Dural et al. 2006
Canada, Berbee et al. 2006
Japan, Sato et al. 2009
Japan, Satopan, Sato et al. 2009
Japan, Sato et al. 2009
USA, Binder et al. 2004
Lithuania, present study
Mexico, Morris et al. 2008
China, Matheny et al. 2007
Mexico, Morris et al. 2009
Brazil, Martin & Raidl 2002
Brazil, Martin & Raidl 2002
China, Matheny et al. 2007
Brazil, Martin &Raidl 2002
Netherlands, den Bakker et al. 2004
Netherlands, den Bakker et al. 2004
Japan, Sato et al. 2009
USA, Wu et al. 2000
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

207
vate, yellowish in KOH, 4-spored, a few 2-spored. Pl e u r o c ys t i d i a
(82.9) 84.2 ± 11.5 (85.8) x (15.6) 18.8 ± 2.5 (23.4) µm (n = 16), ventricose
to subclavate, smooth, readily collapsing (Fig. 5).
H ab i t a t . On soil, solitary to scattered under pines (Pinus mugo
and P. sylvestris), occasionally abundant in P. mugo plantations.
D is t r i b u t io n i n L i t h u a ni a . So far, the fungus was ob-
served only in the Curonian Spit; no records for the continental part of
the country.
M a t e r i a l e x a m i n e d . LITHUANIA, Curonian Spit National Park,
Smiltyne˙ forest district, forest square No 22, area No 27, N55�38’024” E21�07’652”,N55�38’024” E21�07’652”,�38’024” E21�07’652”,
among mosses under P. sylvestris, 5 Sep 2007, leg. J. Kasparavicˇius (BILAS 48763);
Same locality and collector, 3 Sep 2008 (BILAS 48767); 7 Sep 2009 (BILAS 48764);
6 Oct 2009 (BILAS 48768); forest square No 22, area No 24, N55�38’074” E21�07’679”,N55�38’074” E21�07’679”,�38’074” E21�07’679”,
among mosses under P. mugo, 7 Sept 2009, leg. J. Kasparavicˇius (BILAS 48765 and
48766). Note: additional observations of the fungus (specimens not collected) were
made: Curonian Spit National Park, Smiltyne˙ forest district, forest square No 22,
N55�37’877” E21�07’582”, among mosses under�37’877” E21�07’582”, among mosses under P. mugo (massive appearance of ba-
sidiomata), 1 Sep 2010 (observed by J. Motieju¯ naite˙); Juodkrante˙ forest district, for-
est square No 54, N55�31’029” E21�06’297”, among mosses underN55�31’029” E21�06’297”, among mosses under�31’029” E21�06’297”, among mosses under P. sylvestris (sev-
eral solitary fruiting bodies), 2 Sep 2010 (observed by J. Motieju¯ naite˙).
Phylogenetic analysis
Four ITS sequences were produced in this study, each consisting
of 692 characters. They were added to 43 sequences of boletoid species
of the genera Boletus, Boletellus, Aureoboletus, Boletinellus, Retibo-
letus and Leccinum) (Tab. 1, Fig. 6). The sequence of Suillus america-
nus was used as outgroup. Sequences of the Lithuanian isolates nested
in a Boletus s. str./Boletellus clade, subclade of smooth-spored Bo-
letellus (section Mirabilis) (Fig. 6). They formed a discriminatory group
with B. projectellus with a high bootstrap value (94 %), thus substan-
tiating morphological and anatomical identication of the species.
Discussion
The genus Boletellus Murill encompasses ca. 50 described species
worldwide, the majority with tropical distributions (Ortiz-Santana et
al. 2007, Fulgenzi et al. 2008, Halling & Ortiz-Santana 2009). Boletel-
lus has been variably dened, yet all denitions have the following
features in common: spore deposit olivaceous brown, hymenophore
yellow becoming olivaceous with time, and a boletoid tube trama (Ful-
genzi et al. 2008). Singer (1986), e. g., accepted a wide denition of the
genus, including species with both smooth and variously ornamented
spores; meanwhile Smith and Thiers (1971) included only the species
with longitudinally striate, winged or ridged spores. There is no ex-
haustive molecular research on the genus, however, the phylogram of
Dentinger et al. (2010, p. 1286) demonstrated that Boletellus (includ-
ing species with both smooth and ornamented spores) formed a sepa-
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

208
rate clade together with Aureoboletus. Boletellus projectellus, with
smooth and extremely large spores – the largest of all boletes (Singer
1945) belongs to the section Mirabilis that encompasses the smooth-
spored species of the genus; its exceptional spore size and morphologi-
cal characters make this fungus fairly easily identiable. In Lithuania
and in the adjacent countries there are no similar boletoid fungi that
may be confused with B. projectellus and, as the region is compara-
tively well studied by mycologists, it is safe to assume that such a con-
spicuous species could not have escaped the attention of researchers or
amateurs. An indirect support for the assumption that B. projectellus
is a recent arrival to the region is the fact that from the Curonian Spit,
so far the only Lithuanian locality of this fungus, a number of new al-
ien fungi were recorded, especially recently (Markovskaja & Treigiene˙
2009, Markovskaja et al. 2011, Markovskaja and Kutorga pers.
comm.).
Members of Boletellus are found in all pantropical regions and, to
some extent, in temperate zones of North America (Pegler & Young
1981) and Asia (Nagasawa 1997). Boletellus projectellus is widely dis-
tributed in eastern North America from Canada to Mexico (Murrill
1938, Coker & Beers 1943, Singer 1945). It is also known from SE Asia,
Taiwan (Chen et al. 1998). Boletellus projectellus is associated with
pines and is apparently capable of switching hosts within the genus
Pinus quite easily (Both 1993), thus making a good candidate for an
invasive species. In Taiwan, however, it was reported from a broad-
leaved forest (the presence of pines was not mentioned) (Chen et al.
1998).
In Lithuania, B. projectellus was recorded in stands of both local
Pinus sylvestris and introduced P. mugo, though, according to our ob-
servations, production of basidiomata was obviously more abundant
in the latter stands. It is difcult to tell when and how the species ar-
rived to the area. Interviews with local people have revealed that rst
basidiomata were observed in the eighties of the 20th century and that
they remained very much localized and scarce approximately for a
decade. The fungus started to spread during the last decade, and at
present it is one of the more common boletoid mushrooms in the north-
ern part of the Curonian Spit, though it has not yet spread to the con-
tinental part of the country. Nothing is known about the rates of Bo-
letellus spreading, but Pringle et al. (2009) reported in a well-docu-
mented study that the ectomycorrhizal Amanita phalloi�es (Vaill. ex
Fr.) Link, alien to North America, may spread up to 10 km per year.
Based on such an assumption and the information on the rst sightings
of B. projectellus in the Curonian Spit, it can be estimated that the
fungus arrived about 30-35 years ago.
Figs. 1–5. – Boletellus projectellus: 1. Mature basidioma. 2. Young basidioma in
longitudinal section. 3. Old basidioma. 4. Spores. 5. Cystidium.
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

209
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

210
Fig. 6. – Neighbour-joining tree derived from ITS region of the rRNA gene, calcu-
lated in Lasergene without pairwise corrections. Numbers above branches are boot-
strap values obtained from 1000 replications, shown only for branches supported by
more than 50 %. Length of the branches is proportional of number of changes.
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

211
Though theoretically there may be several possible pathways of
arrival as well as several vectors of introduction, the likeliest way for
a mycorrhizal fungus is through roots and soil of introduced plant
seedlings (Vellinga et al. 2009). The Curonian Spit holds extensive
plantations of alien mountain pine, some of considerable age – the rst
seedlings of the introduced P. mugo were imported from Denmark and
Belgium in the 19th century (Bucˇas 2001). The use of imported seed-
lings, however, was ceased after World War II. Few individuals of oth-
er alien pines were planted in the Curonian Spit at various periods, but
their origins are difcult to track. Other possibilities may be an intro-
duction through human transport (especially marine) or natural ones
like wind or birds. These possibilities, however, would involve disper-
sal via spores, which is effective at a local scale, but not at wide dis-
tances (Vellinga et al. 2009). It is assumed that birds can play a role in
distribution of lichen diaspores, e.g., the appearance and establish-
ment of the arctic alpine lichen Flavocetraria nivalis (L.) Kärnefelt &
A. Thell in Vistula Spit (Poland) is attributed to migrating birds
(Fałtynowicz & Budzbon 1983), but, such assumptions do not exist so
far for mycorrhizal fungi.
This is the rst record of B. projectellus in Europe, and, assuming
that it is an invasive fungus, it will be possible to track and evaluate its
prospective spreading along geographic and climatic gradients. It is
already known that the fungus invades sites harbouring alien plants
and native forest as well. At this stage of invasion it would be worth-
while to study also its inuence and relationships with indigenous my-
corrhizal fungi.
Acknowledgments
The study was funded by a grant No LEK -23/2010 from the Re-funded by a grant No LEK -23/2010 from the Re-
search Council of Lithuania.
References
Berbee M. L., Inderbitzin P., Zhang G. (2001) Class project, University of British
Columbia Biology 323, Structure and Reproduction of Fungi. http://www.
ncbi.nlm.nih.gov/ (Last accessed: 19.04.2011).
Berbee M. L., Wright S. H. A., Lim S. (2006) University of British Columbia Biol.
323 class project. http://www.ncbi.nlm.nih.gov/ (Last accessed: 19.04.2011).
Beugelsdijk D. C. M., van der Linde S., Zuccarello G. C., den Bakker H. C., Draisma
S. G. A., Noordeloos M. E. (2008) A phylogenetic study of Boletus section
Boletus in Europe. Persoonia 20: 1–7.
Binder M., Matheny P. B., Hibbett D. S. (2004) Assembling the Fungal Tree of Life.
http://www.ncbi.nlm.nih.gov/ (Last accessed: 19.04.2011).
Both E. E. (1993) The Boletes of North America. Buffalo Museum of Science, New
York.
Bucˇas J. (2001) Kuršiu˛ Nerijos nacionalinis parkas. Savastis, Vilnius.Savastis, Vilnius.
Chen Ch. M., Yeh K. W., Hsu H. K. (1998) The Boletes of Taiwan (X). Taiwania 43(2):
140–149.
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

212
Coker W. C., Beers A. H. (1943) The Boletaceae of North Carolina. North Carolina
Press, Chapel Hill.
den Bakker H.C., Gravendeel B., Kuyper T.W. (2004) An ITS phylogeny of Leccinum
and an analysis of the evolution of minisatellite-like sequences within ITS1.
Mycologia 96: 102–118.
Dentinger B. T. M. Ammirati, J. F., Both E. E., Desjardin D. E., Halling R. E., Henkel
T. W., Moreau P.-A., Nagasawa E., Soytong K., Taylor A. F., Watling R.,
Moncalvo J.-M., McLaughlin D. J. (2010) Molecular phylogenetics of porcini (2010) Molecular phylogenetics of porciniMolecular phylogenetics of porcini
mushrooms (Boletus section Boletus). Molecular Phylogenetics an� Evolu-
tion 57: 1276–1292.
Dentinger B. T. M., McLaughlin D. J. (2007) The evolution of specialization in ecto-
mycorrhizal fungi: an example from porcini. http://www.ncbi.nlm.nih.gov/
(Last accessed: 19.04.2011).
Durall D. M., Gamiet S., Simard S. W., Kudrna L., Sakakibara S. M. (2006) Effects
of clearcut logging and tree species composition on the diversity and com-
munity composition of epigeous fruit bodies formed by ectomycorrhizal fun-
gi. http://www.ncbi.nlm.nih.gov/ (Last accessed: 19.04.2011).
Fałtynowicz W., Budzbon E. (1983) Drugie stanowisko Cetraria nivalis (L.) Ach. na
niz˙u polskim. Fragmenta Floristica et Geobotanica 29 (3–4): 451–456.
Fulgenzi T. D., Mayor J. R., Henkel T. W., Halling R. E. (2008) New species of Bo-
letellus from Guyana. Mycologia 100: 490–495.
Gudelis V. (1998) Lietuvos i˛ju¯ris ir paju¯ris. In: Lietuvos mokslas 17 (eds. Liekis A.,
Misevicˇius J.), Lietuvos mokslas, Vilnius: 1-444.
Halling R. E., Ortiz-Santana B. (2009) A revision of Boletellus sect. Ixocephali. My-
cological Progress 8: 237–244.
Jones M. D., Twieg B. D., Durall D. M., Berch S. M. (2008) Location relative to a
retention patch affects the ECM fungal community more than patch size in
the rst season after timber harvesting on Vancouver Island, British Colum-
bia. Forest Ecology an� Management 255: 1342-1352.
Kirstukas M. (ed.) (2004) Lietuvos gamta. Lutute˙, Kaunas.
Kornerup A., Wanscher J. H. (1978) Methuen han�book of colour. 3rd edn. Eyre
Methuen Ltd, London.
Kuršiu˛ Nerijos nacionalinio parko direkcija/Apie nacionalini˛ parka˛/Gamta. http://
nerija.am.lt/VI/index.php#r/195 (Last accessed: 25.03.2011).
Leonardi M., Paolocci F., Rubini A., Simonini G., Pacioni G. (2005) Assessment of
inter- and intra-specic variability in the main species of Boletus e�ulis
complex by ITS analysis. FEMS Microbiological Letters 243: 411-416.
Markovskaja S., Kacˇergius A., Treigiene˙ A. (2011) Occurrence of new alien patho-
genic fungus Mycosphaerella �earnessii in Lithuania. Botanica Lithuanica
17(1): 29–37.
Markovskaja S., Treigiene˙ A. (2009) New data on invasive pathogenic fungus Doth-
istroma septosporum in Lithuania. Botanica Lithuanica 15(1): 41–45.
Martin M.P., Raidl S. (2002) The taxonomic position of Rhizopogon melanogas-
troi�es (Boletales). Mycotaxon 84: 221–228.
Matheny P. B., Wang Z., Binder M., Curtis J. M., LimY. W., Henrik Nilsson R.,
Hughes K. W., Hofstetter V., Ammirati J. F., Schoch C. L., Langer E., Langer
G., McLaughlin D. J., Wilson A. W., Froslev T., Ge Z. W., Kerrigan R. W., Slot
J. C., Yang Z. L., Baroni T. J., Fischer M., Hosaka K., Matsuura K., Seidl M.
T., Vauras J., Hibbett D. S. (2007) Contributions of rpb2 and tef1 to the phy-
logeny of mushrooms and allies (Basidiomycota, Fungi). Molecular Phyloge-
netics an� Evolution 43: 430–451.
Morris M. H., Perez-Perez M. A., Smith M. E. Bledsoe C. S. (2008) Multiple species
of ectomycorrhizal fungi are frequently detected on individual oak root tips
in a tropical cloud forest. Mycorrhiza 18: 375–383.
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at

213
Morris M. H., Perez-Perez M. A., Smith M. E., Bledsoe C. S. (2009) Inuence of host
species on ectomycorrhizal communities associated with two co-occurring
oaks (Quercus spp.) in a tropical cloud forest. FEMS Microbiology Ecology
69: 274–287.
Murrill W. A. (1938) New Boletes. Mycologia 30: 520–525.
Ortiz-Santana B., Lodge D. J., Baroni T. J., Both E. E. (2007) Boletes from Belize
and the Dominican Republic. Fungal Diversity 27: 247-416.
Nagasawa E. (1997) A preliminary checklist of the Japanese Agaricales. I. The Bo-
letineae. Reports of the Tottori Mycological Institute 35: 39–78.
Pegler D. N., Young T. W. K. (1981) A natural arrangement of the Boletales, with
reference to spore morphology. Transactions of the British Mycological Soci-
ety 76(1): 103–146.
Pringle A., Adams R. I., Cross H. B., Bruns T. D. (2009). The ectomycorrhizal fungus
Amanita phalloi�es was introduced and is expanding its range on the West
Coast of North America. Molecular Ecology 18: 817–833.
Sato H., Tsujino R., Kurita K., Yokoyama K. (2009) Use of ITS sequences for resolv-
ing species richness, ecological differentiation, and geographical distribution
of macro-fungi. http://www.ncbi.nlm.nih.gov/ (Last accessed: 19.04.2011).
Singer R. (1945) The Boletinae of Florida with notes on extralimital species. Far-
lowia 2(1): 97–141.
Singer R. (1986) The Agaricales in mo�ern taxonomy. 4th edn., Koeltz Scientic
Books, Koenigstein.
Smith A. H., Thiers H. D. (1971) The boletes of Michigan. University of Michigan
Press, Ann Arbor.
Vellinga E. C., Wolfe B. E., Pringle A. (2009) Global patterns of ectomycorrhizal
introductions. New Phytologist 181: 960–973.
White T. J., Bruns T., Lee S., Taylor J. (1990) Amplication and direct sequencing of
fungal ribosomal RNA genes for phylogenetics. In: PCR protocols: a gui�e to
metho�s an� applications (eds. Innis M. A., Gelfand D. H., Sninsky S. S.,
White T. J.), Academic Press, San Diego, USA: 315–322.
Wu Q. X., Mueller G. M., Lutzoni F. M., Huang Y. Q., Guo S. Y. (2000) Phylogenetic
and biogeographic relationships of eastern Asian and eastern North Ameri-
can disjunct Suillus species (fungi) as inferred from nuclear ribosomal RNA
ITS sequences. Molecular Phylogenetics an� Evolution 17: 37–47.
(Manuscript accepte� 11. November 2011; Correspon�ing E�itor: R. Pö�er)
©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at