Post on 01-Aug-2020
COMMENTARY PAPER
Insect-truffle interactions – potential threats to emerging industries?
Aleksandra Rosa-Gruszecka1, Alan C. Gange2, Deborah J. Harvey2, Tomasz Jaworski1 , Jacek
Hilszczański1, Radosław Plewa1, Szymon Konwerski3, Dorota Hilszczańska4
1 Department of Forest Protection, Forest Research Institute, Sękocin Stary, Braci Leśnej 3,
05-090 Raszyn, Poland
2 School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey
TW20 0EX UK
3 Natural History Collections, Faculty of Biology, Adam Mickiewicz University, Umultowska
89, 61-614 Poznań, Poland
4 Department of Forest Ecology, Forest Research Institute, Sękocin Stary, Braci Leśnej 3, 05-
090 Raszyn, Poland
Corresponding author: A.C. Gange, address above
E: a.gange@rhul.ac.uk
Tel. +44(0)1784 443188
Fax +44(0)1784 414224
Word count: 4,401
Running title: Insects and truffles
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ABSTRACT
Truffle harvests are declining in Europe, due to droughts, and this offers an opportunity for
production to be developed in countries such as the UK and Poland, where rainfall tends to be
higher. Drier Mediterranean summers seem to be associated with a decrease in the harvest of
the Périgord truffle (Tuber melanosporum) in Spain, France and Italy. However, other
species, for example the Burgundy truffle (T. aestivum) offer opportunities for production in
the more temperate environments north of the Alps. Truffles across Europe can be infested by
insect larvae, seriously reducing their economic and culinary quality. Here, using a
combination of literature sources and a field survey, we present a commentary on insects
attacking truffles, aiming to highlight those species that could be potential pests in the British
and Polish emergent industries. There is a remarkable disparity in coincidence of records of
insects and truffles in these countries, yet a survey in Poland confirms that insects can be
abundant. We discuss reasons for this disparity and suggest that biochemical methods could
easily be developed for detection of the truffles and their attackers.
Introduction
Truffles are hypogeous fungi belonging to the Pezizales, mostly in the genus Tuber, and
comprise a large group of ectomycorrhizal fungi growing in symbiosis with the roots of
several vascular plant species (angiosperms and gymnosperms). The fruit body of these fungi
is a subterranean complex apothecium, commonly known as the truffle. The geographic
distribution of truffles mainly covers the temperate zones of the northern hemisphere, with at
least three areas of genetic differentiation in Europe, South East Asia and North America
(Pomerico et al., 2006). So far, seven species of Tuber have been reported from Poland,
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namely T. mesentericum (Ławrynowicz, 1999), T. aestivum, T. excavatum, T. rufum
(Hilszczańska et al., 2008), T. maculatum (Ławrynowicz, 2009), T. macrosporum
(Hilszczańska et al., 2013) and T. brumale (Merényi, et al., 2014). According to the Fungal
Records Database of the British Isles (http://www.fieldmycology.net/FRDBI/FRDBI.asp) all
7 species have been recorded in the UK, though none have many attributed records. That with
the most is T. aestivum with 110 records, but to put this into perspective, the fungal species
with the most records (Hypholoma fasciculare) has 16,259 (as of 21 September 2016).
Economically, truffles are the most valuable non-timber products of forest ecosystems,
and are highly prized for their culinary qualities in countries such as France, Italy and Spain.
Highly desirable truffles (i.e. T. magnatum (white) or T. melanosporum (black)) may attract
remarkable prices, of around €2,000 - €3,000 kg-1, with the industry in Italy worth around
€400 million per annum (Büntgen et al. 2012; Pieroni, 2016). This may be the primary reason
why a truffle industry is emerging in countries such as the UK and Poland. However, it may
also be due to the decline of harvests of the highly-prized black truffle (T. melanosporum) in
its main habitats due to increased frequency of droughts (Büntgen et al., 2011; 2012; 2015).
Although neither T. magnatum nor T. melanosporum have been found in the UK or Poland,
two of their species (T. aestivum and T. brumale) are commercially traded in countries such as
Spain and Hungary (Martin-Santafe et al., 2014; Stobbe et al 2013). Indeed, recent evidence
suggests that T. aestivum in particular may be found in suitable areas north of the Alps, such
as Germany, and even as far north as southern Sweden and Finland (Stobbe et al., 2012;
2013). The first cultivated specimen of T. aestivum was found in England in March 2015
(http://www.bbc.co.uk/news/science-environment-31826764 ) and in Wales in July 2016
(http://www.itv.com/news/wales/2016-07-25/first-ever-cultivated-truffle-harvested-in-wales/).
In Poland, three truffle orchards (with T. aestivum) have been established and maintained by
the Forest Research Institute, one of which is productive (Hilszczańska et al., 2008;
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Hilszczańska and Sierota, 2010, Hilszczańska, 2016). Therefore, there is great economic
potential for the emergent industries in more northerly countries to fill gaps in the European
market, and it is timely to identify any problems that might reduce their potential in future.
Ecologically, these fungi are of considerable importance because of the benefits of the
mutualistic association they provide to their host plants (Pacioni and Comandini, 1999). In
addition, the relatively long-lived fruit body provides a food source for invertebrates and
vertebrates (Johnson, 1996; Blackwell, 2005). Some species of truffles, e.g. T. magnatum, T.
melanosporum and T. aestivum, have the high culinary value because of their aroma (Mello et
al., 2006) and in natural habitats, the volatiles produced are essential for attracting animals
that spread the spores (Fogel and Trappe, 1978). However, some animals have evolved a
capacity for feeding on truffle sporophores. These belong to various taxonomic groups and
are termed ‘hydnophagous’ (Pacioni, 1989), from Greek ‘hydnon’, truffle, and ‘phagous’,
eating. For certain animals, such as mammals (rodents, deer, boars), birds and slugs, truffles
are a valuable part of the diet (Johnson, 1996; Vernes et al. 2015), yet for several species or
genera of Arthropods, mainly in the Coleoptera and Diptera, the fungi may represent their
complete diet (Pacioni et al., 1995; DiSanto, 2013). Indeed, there is much anecdotal evidence
that truffles may be discovered by searching for the flies that oviposit within the fruit body,
though this is a laborious and unpredictable procedure (Blackwell, 2005). Scattered through
the literature are occasional reports of truffle sporocarps being infested by insect larvae, thus
reducing their marketable value considerably (Ciampolini and Suss, 1982; Martin-Santafe et
al., 2014). Our aim here is to provide a survey of the insects that may be associated with
truffles in two countries, which as well as being ecologically interesting, highlights potential
problems for the emerging truffle industry.
Survey methods
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We present a survey of known host associations in each country, using databases and field
sampling. The UK insect fauna is relatively well recorded and we used national distribution
data, available through the National Biodiversity Network Gateway (https://data.nbn.org.uk/).
The UK National Grid divides the country into 2,500 10 km x 10 km squares and records are
provided at this scale. We extracted a list of the 10 km x 10 km squares in which T. aestivum
has been recorded and compared these with 10 km x 10 km records of the principal insect
species associated with truffles (described below).
National record data for Poland are far less developed, but we extracted data for fungi and
insects from the Universal Transverse Mercator (UTM) 10 km x 10 km grid
(http://baza.biomap.pl/pl/db). The Polish UTM divides the country into 3,384 10 km x 10 km
squares (Iwan et al., 2012). These were supplemented by surveys in four geographical regions
in Poland: Nida Basin, Przedbórz Upland, Miechów Upland and Chełm Hills (Table 1).
Records were obtained using two methods of collecting insects associated with truffles. In
2012-2014 Tuber spp. fruit bodies inhabited by adults and larvae were collected and larvae
reared to adult. Information on sampling effort is given in Table 2.
In addition, insects were also collected in two adjacent regions of Nida Basin and
Miechów Upland with traps installed in natural habitats of T. aestivum. A total of 24 modified
funnel traps were used in both localities. To minimize collection of non-target invertebrates
(e.g. Carabidae, Silphidae) or small vertebrates (lizards, mice), traps were buried in soil with
the upper edge of the funnel left a few centimeters above the ground, and were covered with a
plastic roof placed 2 cm above the funnel. To preserve collected insects, a container filled
with 200 ml of ethylene glycol was put underneath the funnel. Each trap was baited using a
small piece of T. aestivum mature fruit body, placed inside a 2 ml perforated container that
was installed under the trap cover. Traps were emptied every third week from mid-July to
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early October 2012. Beetles were identified by S. Konwerski and flies (Diptera) were
identified by A. Woźnica.
Truffle insects and their distributions
The coleopteran fauna associated with truffles is mainly represented by the beetle Leiodes
cinnamomea (Coleoptera: Staphylinoidea) (Arzone, 1970; 1971). Adult females of the species
are attracted by truffle volatiles in its early stage of growth, but not when the fruit body is
mature (Hochberg et al., 2003). The beetle appears to be specific to the genus Tuber,
particularly T. melanosporum, with some records from T. aestivum and T. excavatum (Fogel
and Peck, 1975; Pacioni et al., 1991; Bratek et al., 1993) and completes its life cycle in or
adjacent to the fruit body (Arzone, 1970; Newton, 1984). Nevertheless it can cause extensive
damage to the truffle (Fig. S1). In the UK, L. cinnamomea is considered hard to find
(Blackwell, 2005) and is designated as Nationally Notable, having been recorded in only 25
of the 2,500 10 km x 10 km squares of the National Grid (https://data.nbn.org.uk/ (accessed
21/9/2016)). Its most likely host in the UK, T. aestivum, has been recorded from 40 of the 10
km x 10 km squares, yet in only 3 squares are there coincidental records of beetle and truffle.
This represents just 12% of the recorded beetle distribution and 7.5% of squares with truffle
records. Given that the insect is host specific, one would expect that all beetle records would
coincide with those of the truffle, but instead there is a great discrepancy in records. This is
likely due to the well-known bias that exists within such data bases (e.g. Ward, 2014), but
with truffles in a country like the UK it is likely to be particularly acute. As the fungus is
highly prized, locality records are very unlikely to be advertised by those seeking the fruit
bodies for commercial purposes, creating a highly biased distribution. Entomologists are more
likely to present their records, as they are far less likely to have a vested interest in truffle
collection. Thus, the available data suggest that the insect may have little impact on the
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industry, but this is subject to the serious bias aforementioned. It is likely that L. cinnamomea
is considerably more common than thought and should not be dismissed as a potential pest
species.
In Poland L. cinnamomea is known from 15 squares of the UTM 10 km x 10 km grid
(http://baza.biomap.pl/pl/db (accessed 21/9/2016)). However, none of these localities are
coincident with known Tuber spp. sites. The field survey (Table 1) showed that larvae of this
beetle can infest ascocarps of T. aestivum and T. exacavatum. However, considerably fewer
larvae were found in T. excavatum. Along with L. cinnamomea another leiodid beetle, L.
oblonga, was found, both as adults and larvae, in T. aestivum fruit bodies. Other hosts of this
species’ larvae were T. excavatum and T. rufum (Table 1). Koch (1991) reported L. oblonga
from truffles, but without distinguishing the fungal species. L. oblonga has been recorded
from 16 10 km x 10 km squares in the UK, yet only one of these coincides with T. aestivum.
The results here seem to be the first report on L. oblonga associations with truffles. L. oblonga
is very similar to L. cinnamomea but correct identification of both species is possible based on
the analysis of the male genitalia (Nunberg 1987). Therefore, further studies on Leiodidae
associated with truffles should take into consideration the problem of similarity of both
species and molecular methods may be best employed to separate them.
Other species of beetle in the same family as L. cinnamomea in the UK include
Agaricophagus cephalotes and Colenis immunda (Horsfield, 2002). While these species are
known to attack T. aestivum elsewhere (Bratek et al., 1993; 2010), there is no information
available on their biology in the UK. A. cephalotes is very rare (just seven 10 km x 10 km
records), while C. immunda is more common (57 10 km x 10 km records, of which three
coincide with T. aestivum). C. immunda was also reared from fruit bodies in the Polish field
survery (Table 1). However, this beetle appears to have many above-ground fungal hosts
(Schigel, 2011), and so is likely to pose little threat to truffle cultivation.
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The fruit bodies of truffles are also commonly inhabited by various species of Diptera,
mainly of the genus Suillia (Ciampolini and Suss, 1982; Krivosheina, 2008). The adult
females fly close to the soil surface and lay their eggs on the ground, above the truffle fruit
bodies, so that the larvae can easily reach them on hatching (Talou et al., 1990). Larval
feeding can cause extensive damage (Fig. S2). Martin-Santafe et al. (2014) and Duaso (2012)
report infestation of fruit bodies of T. aestivum and T. melanosporum by another fly,
Helomyza tuberivora (= Suillia gigantea) but this species does not seem to occur in the UK.
Instead, Suillia affinis (which appears to be a senior synonym of Helomyza affinis) and S.
pallida do occur, S. affinis having been recorded from 99 of the 10 km x 10 km squares and S.
pallida from 58. Of the S. affinis squares, just four are coincidental with T. aestivum,
suggesting that either the same biased recording problem exists or that the fly is not host
specific (Baehrmann and Adaschkiewitz, 2003). Notwithstanding the recording problem, the
four squares represent 5% of fly squares and 12.5% of truffle squares. A similar situation
exists with S. pallida where the figures are 3.4% and 5%, respectively. Thus neither species
may pose much of a threat to UK truffle species such as T. aestivum, even though they infest
these elsewhere (Papp, 1994; Baehrmann and Adaschkiewitz, 2003).
As in UK, the truffle fly S. gigantea has not been recorded in Poland, in contrast to S.
affinis which was reported from the southern part of the country (Ojców National Park)
(Woźnica and Klasa, 2009). Soils in this part are calcareous and conducive to truffle
development, so T. aestivum could be a potential host species. The fly was the most numerous
of all trapped insects (Table 1). Based on research of Lo Giudice and Woźnica (2013) we can
also speculate that since localities of S. gigantea and S. affinis overlap in such regions as
Tuscany, Lombardia and Umbria, S. affinis may be an indicator of truffle presence and a
potential pest in Poland.
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Chandler (2010) reported the association with Tuber for Diptera of the genus Cheilosia
(Syrphidae) and one species (C. soror) has been bred from truffles (Falk, 1991). This fly has
been recorded from 173 of the 10 km x 10 km squares in the UK. Perhaps of most interest is
that this species shows a very different coincidence with T. aestivum compared with the
previous insect species: 35% of the truffle squares also possess a record of this fly. While
there is virtually no data available on the biology of this species, it would certainly merit
investigation as a species of potential pest concern.
Truffle biochemistry
The chemicals given off by truffles contribute to their characteristic aroma and gives them
their high monetary value (Costa et al., 2015). This suite of volatiles gives each a species-
specific profile, which changes over time. This has been linked to maturation of the fruit
body, as well as environmental factors and more recently to genetic variability (Splivallo et
al., 2012). A single fruit body produces 20-50 volatile organic compounds (Culleré et al.,
2010; Splivallo et al., 2011), but those considered to be biologically important include
dimethyl sulphide, which attracts dogs and pigs, and eight carbon-containing volatiles, which
attract the two main insect species associated with truffles, L. cinnamomea and S. pallida
(Talou et al., 1990; Splivallo et al., 2012).
Demand for such a rare and expensive crop in the UK and Poland can only be realised if
the truffles are both of good quality and pest-free. Recent chemoecological research has,
therefore, centred around identification of the exact chemical profiles emitted by the fruit
bodies and how they change over time. This information can then be used in the development
of devices to monitor both the quality of the truffles i.e. electronic noses (Costa et al., 2015;
Pennazza et al., 2013) and pest control in truffle harvests (Hochberg et al., 2003).
9
The use of volatiles given off by the fruit bodies of truffles to reduce infestation by insect
species has engendered remarkably little interest. To date, this work has focused on L.
cinnamomea, and its attraction to both the volatiles produced by truffles and those produced
by the beetle itself (Hochberg et al., 2003). These authors showed that neither sex is attracted
by ripe truffle odours, but that females are attracted to immature truffles and males to
pheromones produced by females. To date, these observations have never been examined
experimentally. It could be critical in aiding the emergent industries to protect their harvests
against pest insects through the development of attractant traps that would divert the beetles
from the fruit bodies. However, this presents an intriguing philosophical dilemma; if the
beetles are as rare as national records suggest, then one could question the ethics of
developing traps that kill such rare, pest insects. There is a clear need to develop traps that
catch live insects, in order to accurately determine population sizes and distributions and
thereby address this dilemma.
Conclusions and future perspectives
Truffles are highly prized and their economic value is dependent not only on the aromas they
emit, but also on the fruit bodies being free of insect larvae. Truffle harvests have shown
notable declines in parts of Europe, and this offers important economic opportunities in
countries such as the UK and Poland to fill market gaps. It is important that these emergent
industries do not fail due to poor material that is infested with insects. Here, we have tried to
highlight those species of insect most likely to become pests in this industry. Understanding
their ecology will enable us to determine whether the disparity in national records of insects
and their hosts is real or due to recorder bias. Prevention of attack by insects is important for a
truffle producer, but presents an ethical dilemma of whether such rare insects should be
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trapped and killed. The development of electronic noses could aid truffle harvesters, since
these will be consistent and immortal, unlike a pig or a dog.
Acknowledgments
We are deeply grateful to Dr Domizia Donnini (University of Perugia, Italy) for help with
specimens of Leiodes cinnamomea, Dr Andrzej Woźnica (Wrocław University of
Environmental and Life Sciences, Poland), Dr Cezary Bystrowski (Forest Research Institute,
Poland) for identification of Suilla affinis and Karol Komosiński (University of Warmia and
Mazury, Poland) for identification of Atheta dilaticornis. We also thank prof. Jerzy Borowski
(Warsaw University of Life Sciences, Poland) for providing literature.
This work was supported by State Forest Holding, project No. OR-2717/19/11, Forest
Research Grant No. 260102 and by the Polish Ministry of Science and Higher Education,
project No. 240309.
References
Arzone, A., 1970. Reperti ecologici ed etologici di Leiodes cinnamomea Panzer vivente su
Tuber melanosporum Vittadini (Coleoptera Staphylinoidea). Annali della Facoltà di
Sciènze Agrarie della Università Degli Studi di Torino, 5: 317-357. [in Italian]
Arzone, A., 1971. Nuovi reperti sulla biologia di Leiodes cinnamomea Panzer in Tuber
magnatum Pico (Coleoptera, Staphylinoidea). Allionia: Bollettino dell'Instituto ed Orto
Botanico dell'Universitaà di Torino, 17: 121-129. [in Italian]
Baehrmann, R., Adaschkiewitz, W., 2003. Beitrag zur Oekologie und Fauna der
Heleomyzidae Mitteldeutschlands (Insecta: Diptera). Faunistische Abhandlungen
(Dresden) 24: 185-204. [in German]
11
Blackwell, T., 2005. Discovering Discos and other Ascomycetes. Field Mycology 6, 15-21.
Bratek, Z., Papp, L., Merkl, O., Takács, V., 1993. Föld alatti gombákon élő rovarok.
Mikológiai Közlemények, 31, 55-65. [in Hungarian]
Bratek, Z., Merenyi, Z., Illyes, Z., Laszlo, P., Anton A., Garay, J., et al., 2010. Studies on the
ecophysiology of Tuber aestivum populations in the Carpatho-Panonian region. Austrian
Journal of Mycology 19, 221-226.
Büntgen, U., Tegel, W., Egli, S., Stobbe, U., Sproll, L., Stenseth N.C. 2011. Truffles and
climate change. Frontiers in Ecology and the Environment 9, 150-151.
Büntgen, U., Egli, S., Camarero, J.J., Fischer, E.M., Stobbe, U., Kauserud, H. et al., 2012.
Drought-induced decline in Mediterranean truffle harvest. Nature Climate Change 2, 827-
829.
Büntgen, U., Egli, S., Schneider, L., von Arx, G., Rigling, A., Camarero, J.J., et al., 2015.
Long-term irrigation effects on Spanish holm oak growth and its black truffle symbiont.
Agriculture, Ecosystems & Environment 202, 148-159.
Ciampolini, M., Suss L., 1982. Nuovi reperti sulla mosca dell'aglio, Suillia univittata (von
Roser) (Diptera Heleomyzidae). Bollettino di Zoologia Agraria e di Bachicoltura, 17, 19-
38. [in Italian]
Chandler, P., 2010. A Dipterist's Handbook (2nd edition). The Amateur Entomologists'
Society, pp. 424-439.
Costa, R., Fanali, C., Pennazza, G., Tedone, L., Dugo, L., Santonico, M. et al., 2015.
Screening of volatile compounds composition of white truffle during storage by GCxGC-
(FID/MS) and gas sensor array analyses. LWT – Food Science and Technology 60, 905-
913.
Culleré, L., Ferreira, V., Chevret, B., Venturini, M.E., Sánchez-Gimeno, A.C., Blanco, D.,
2010. Characterisation of aroma active compounds in black truffles (Tuber
12
melanosporum) and summer truffles (Tuber aestivum) by gas chromatography–
olfactometry. Food Chemistry 122, 300–306.
Di Santo, P., 2013. Interazioni pianta–micorriza–insetto: il modello Quercus sp.–Tuber sp.–
Leiodes cinnamomea (Panzer). Dottorato di Ricerca in “Difesa e Qualita delle Produzioni
Agro-Alimentari e Forestali”, Universita degli Studi del Molise [in Italian].
Duaso, L.C., 2012 Artrópodos parásitos asociados a carpóforos del genre Tuber. PhD Thesis,
University of Zaragoza. [in Spanish].
Falk, S.J., 1991. A Review of the Scarce and Threatened Flies of Great Britain Part 1.
Research and Survey in Nature Conservation no. 39. JNCC, Peterborough.
Fogel, R., Peck, S.B., 1975. Ecological Studies of Hypogeous Fungi I. Coleoptera associated
with sporocarps. Mycologia 67, 741-747.
Fogel, R., Trappe, J.M., 1978. Fungus consumption (mycophagy) by small animals.
Northwest Science 52, 1–31.
Hilszczańska, D., 2016. Polskie trufle - skarb odzyskany. O hodowli i kulinariach
podziemnego przysmaku. Centrum Informacyjne Lasów Państwowych, Warszawa, 66 pp.
ISBN 978-83-63895-87-7 [in Polish].
Hilszczańska, D., Sierota, Z., Palenzona, M., 2008. New Tuber species found in Poland.
Mycorrhiza 18, 223-226.
Hilszczańska, D., Sierota, Z., 2010. First attempt towards cultivation of Tuber aestivum in
Poland. Austrian Journal of Mycology 19, 209-212.
Hilszczańska, D., Rosa-Gruszecka, A., Sikora, K., Szmidla, H. 2013. First report of Tuber
macrosporum occurrence in Poland. Scientific Research and Essays 7, 1096-1099.
Hochberg, M.E., Bertault, G., Poitrineau, K., Janssen, A., 2003. Olfactory orientation of the
truffle beetle, Leiodes cinnamomea. Entomologia Experimentalis et Applicata 109, 147-
153.
13
Horsfield, D., 2002. Annotated keys to the British Leiodidae, a correction. Entomologist’s
Monthly Magazine 138, 1652-1655.
Iwan, D., Kubisz, D., Tykarski, P., 2012. Coleoptera Poloniae: Tenebrrionoidea
(Tenebrionidae, Boridae). Critical checklist, distribution in Poland and meta-analysis.
University of Warsaw – Faculty of Biology, Natura optima dux Foundation, Warszawa,
480 pp.
Johnson, C.N., 1996. Interactions between mammals and ectomycorrhizal fungi. Trends in
Ecology and Evolution, 11, 503-507.
Koch, K., 1991. Die Käfer Mitteleuropas. Ökologie. Goecke & Evers. Krefeld, 2. 382 pp. [in
German]
Krivosheina, N.P., 2008. Macromycete fruit bodies as a habitat for Dipterans (Insecta,
Diptera). Entomological Review 88, 778-792.
Lo Giudice, G., Woźnica A.J., 2013. An updated checklist of the Italian Heleomyzidae
(Diptera: Sphaeroceroidea). Genus 24, 439-458.
Ławrynowicz, M., 1999. Tuber mesentericum, an interesting species of black truffle in
Poland. Acta Mycologica 3, 169-172.
Ławrynowicz, M., 2009. Four Tuber species accompanying T. mesentericum in natural sites
in Poland. Anales del Jardín Botánico de Madrid 66, 145-149.
Martin-Santafe, M., Perez-Fortea, V., Zuriaga, P., Barriuso J., 2014. Phytosanitary problems
detected in truffle cultivation in Spain. Forest Systems 23, 307-316.
Mello, A., Murat, C., Bonfante, P., 2006. Truffles: much more than a prized and local fungal
delicacy. FEMS Microbiology Letters 260, 1-8.
Merényi, Z., Varga, T., Geml, J., Orczán, Á.K., Chevalier, G., Bratek, Z., 2014. Phylogeny
and phylogeography of the Tuber brumale aggr. Mycorrhiza 24, 101-113.
14
Newton, A.F., 1984. Mycophagy in Staphylinoidea (Coleoptera). Fungus–Insect
Relationships. Perspectives in Ecology and Evolution (ed. by Q Wheeler & M Blackwell),
pp. 302-353. Columbia University Press, New York.
Nunberg, M., 1987. Klucze do oznaczania owadów Polski. Część XIX. Chrząszcze –
Coleoptera. Grzybinki – Leiodidae. 60 pp. Warszawa (PWN). [in Polish]
Pacioni, G., 1989. Biology and ecology of the truffles. Acta Medica Romana 27, 104-117.
Pacioni, G., Comandini, O., 1999. Tuber. In: Cairney, J.W.G., Chambers, S.M. (eds).
Ectomycorrhizal fungi. Key genera in profile. Berlin Heidelberg (Springer), pp. 163-186.
Pacioni, G., Bologna, M.A., Laurenzi, M., 1991. Insect attraction by Tuber: a chemical
explanation. Mycological Research 95, 1359-1363.
Pacioni, G., Ragnelli A.M., Miranda M., 1995. Truffle development and interactions with the
biotic environment. Molecular aspects. In: Stocchi V., Bonfante P., Nuti M. (eds.)
Biotechnology of Ectomycorrhizae. Molecular approaches. Plenum Press, New York, pp
213-227.
Papp, L., 1994. Morphology of third instar larva and puparium of three Heleomyzid species
(Diptera: Heleomyzidae). Acta Zoologica Academiae Scientiarum Hungaricae 40, 219-
229.
Pennazza, G., Fanali, C., Santonico, M., Dugo, L., Cucchiarini, L., Dacha, M. et al., 2013.
Electronic nose and GC-MS analysis of volatile compounds in Tuber magnatum Pico:
Evaluation of different storage conditions. Food Chemistry 136, 668-674.
Pieroni, A., 2016. The changing ethnoecological cobweb of white truffle (Tuber magnatum
Pico) gatherers in South Piedmont, NW Italy. Journal of Ethnobiology and
Ethnomedicine 12 (18), DOI 10.1186/s13002-016-0088-9.
Pomerico, M., Figliuolo, G., Rana, G.L., 2006. Tuber spp. biodiversity in one of the
southernmost European distribution areas. Biodiversity and Conservation 16, 3447-3461.
15
Schigel, D., 2011. Polypore-beetle associations in Finland. Annales Zoologici Fennici 48,
319-348.
Splivallo, R., Ottonello, S., Mello, A., Karlovsky, P., 2011. Truffle volatiles: from chemical
ecology to aroma biosynthesis. New Phytologist 189, 688–699.
Splivallo, R., Valdez, N., Kirchoff, N., Castiella Ona, M., Schmidt, J-P., Feussner, I., et al.,
2012. Intraspecific genotypic variability determines concentrations of key truffle volatiles.
New Phytologist 194, 823-835.
Stobbe, U., Büntgen, U., Sproll, L., Tegel, W., Egli, S., Fink, S., 2012. Spatial distribution
and ecological variation of re-discovered German truffle habitats. Fungal Ecology 5, 591-
599.
Stobbe, U., Egli, S., Tegel, W., Peter, M., Sproll, L., Büntgen, U., 2013. Potential and
limitations of Burgundy truffle cultivation. Applied Microbiology and Biotechnology 97,
5215-5224.
Talou, T., Gaset, A., Delmas, M., Kulifaj, M., Montant, C., 1990. Dimethyl sulphide: the
secret for black truffle hunting by animals? Mycological Research 94, 277-278.
Vernes, K., Cooper, T., Green, S. 2015. Seasonal fungal diets of small mammals in an
Australian temperate forest ecosystem. Fungal Ecology 18, 107-114.
Ward, D.F., 2014. Understanding sampling and taxonomic biases recorded by citizen
scientists. Journal of Insect Conservation 18, 753-756.
Woźnica, A.J., Klasa, A., 2009. Heleomyzid flies of the Ojców National Park, with notes on
Suillia lineitergum (Pandellé, 1901) – a species new to the fauna of Poland (Diptera:
Heleomyzidae). Fragmenta Faunistica 52, 181-190.
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Table 1. Insects associated with Tuber spp. in Poland (asterisks indicate number of specimens
reared from fruit bodies of Tuber, data without asterisks show number of specimens caught in
traps).
Insect species Tuber species Nida Basin
Miechów Upland
Przedbórz Upland
Chełm Hills
DipteraHeliomyzidaeSuillia affinis (Meigen, 1830) T. aestivum 2635 1875 3*
ColeopteraBolboceratidaeOdonteus armiger (Scopoli, 1772) T. aestivum 1LeiodidaeAnisotoma orbicularis (Herbst, 1792) T. aestivum 1Apocatops nigrita (Erichson, 1837) T. aestivum 1Colenis immunda (Sturm, 1807) T. aestivum 4* 1Fissocatops westi (Krogerus, 1931) T. aestivum 6 5Leiodes cinnamomea (Panzer, 1793) T. aestivum 33* 25*
T. excavatum 5*Leiodes oblonga (Erichson, 1845) T. aestivum 14+1* 10 22*
T. excavatum 1*T. rufum 7*
Leiodes polita (Marsham, 1802) T. aestivum 2Nargus velox (Spence, 1815) T. aestivum 1Ptomaphagus sericatus (Chaudoir, 1845) T. aestivum 141 3Ptomaphagus varicornis (Rosenhauer, 1847) T. aestivum 1Sciodrepoides fumatus (Spence, 1815) T. aestivum 2 1Sciodrepoides watsoni (Spence, 1815) T. aestivum 5 4PhalacridaeStilbus testaceus (Panzer, 1797) T. aestivum 7*StaphylinidaeAtheta dilaticornis (Kraatz, 1856) T. aestivum 1*
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Table 2. Number of Tuber spp. fruit bodies collected in Poland in 2012-2014.
Tuber species YearRegion
Nida Basin Miechów Upland
Przedbórz Upland Chełm Hills
T. aestivum2012 35 32013 125 1662014 484 368
T. excavatum2012 2362013 4272014 291 17
T. rufum2012 120132014 5
Total 1599 537 22
18
Supplementary on-line material
Insect-truffle interactions – potential threats to emerging industries?
Aleksandra Rosa-Gruszecka, Alan C. Gange, Deborah J. Harvey, Tomasz Jaworski , Jacek
Hilszczański, Radosław Plewa, Szymon Konwerski, Dorota Hilszczańska
Fig S1 Damage to T. aestivum fruit bodies by larvae and adults of L. cinnamomea
Fig S2 Extensive damage to T. aestivum caused by Suillia larvae.