Truffle diversity and habitats in Balkan Penninsula and Pannonian region

Dr Zaklina Marjanovic Institute for Multidisciplinary Research, University of Belgrade, Kneza Viseslava 1, 11030 Belgrade, Serbia,

The habitats have been widely described from traditional regions for centuries

• Italy, hosting all European commercial truffle species, made magnatum famous as Tartufo Bianco D’alba or “Piedmont” • France has always been famous for truffe de Burgogne, or truffe du Périgord

• The stories of have been built in these two countries through many years of gastronomic history, with devotion and special attention

• However, in past 20 years, the science and practice of truffles has grown enormously, providing much more realistic picture on their real occurrences in Europe

The market has drastically changed

• Spain has experienced enormous development in T. melanosporum plantations

• T.aestivum/uncintum has been detected in almost all countries of Europe, but serious change in the market came with discovery of extremely productive areas in Hungary and eastern Balkan countries

• T . magnatum is collected in Hungary and probably all Balkan countries, with the most significant amounts coming from Serbia and probably Romania (no official records)

• It became clear that Balkan Peninsula and Pannonian region are probably different kind of truffle paradise

Focus on Balkan peninsula and neighbouring regions Bonito et al 2013

We estimated that Tuber diverged from other genera in the early Cretaceous 156 Mya, and by end of the Cretaceous (65 Mya) most of its extant subgeneric lineages were present.

Extant species in a number of Tuber clades can associate with angiosperms, Pinaceae, and parasitic orchid monocots (e.g. T. aestivum). Character state reconstructions indicate the most recent common ancestor to the was likely an ectomycorrhizal symbiont of angiosperms.

Based on our analyses, alveolate-reticulate ornamentation is the plesiomorphic (ancestral) condition for Tuber When T. Aestivum ancestor and T. magnatum were diversifying in Europe according to Bonito et al 2013

The fossil evidence suggests a predominantly tropical to subtropical climate throughout the Late Cretaceous. The first Late Cretaceous angiosperms were mostly weedy, pioneering shrubs. These were likely to have been of disturbed habitats in seasonally dry tropical or subtropical climates. The later angiosperms from the same period had long, slender leaves suggesting a predominantly tropical climate. http://palaeo.gly.bris.ac.uk/communication/willson/palaeoevidence.html Eocene

T. aestivum T. melanosporum

"Late Paleocene Thermal Maximum", was a time period with more than 5–8 °C warmer global average temperature rise across the even. Oligocene Very warm and humid climate, mostly coniferous ECM forests, only in the mountains, in lowlands AM forests…

App. 15-13 MY ago… Last ice age map of Europe (from Bohn et al 2014) Refugiums for all European plants during ice ages

Nowadays… Climate types in Europe (from Bohn et al 2014): IV Mediterranean; VI Continental ; VII dry steppe Serbia land use map from EEA

Fruska gora Djerdap

Tara vegetation map of Balkans

Illyrian Mesian vegetation vegetation province province True truffles of Serbia (13 confirmed species + 3 so far moleculraly unconfirmed)

• Tuber puberulum Berk. & Broome • Tuber rufum Pico f. rufum • Tuber rufum var. apiculatum E. Fisch. • Tuber rufum var. ferrugineum (Vittad.) Montecchi & Lazzari • Tuber rufum var. lucidum (Bonnet) Montecchi & Lazzari • Tuber rufum var. nitidum (Vittad.) • Tuber aestivum Vittad. Montecchi & Lazzari

• Tuber borchii Vittad. • Tuber petrophilum (Milenkovic et al 2015) • Tuber brumale Vittad.s.l. • Tuber dryophyllum Vitt, • Tuber excavatum Vittad. • Tuber foetidum Vittad. • Tuber fulgens Quél. • Tuber macrosporum Vittad. • Tuber maculatum Vittad. • Vittad. • Tuber mesentericum Vittad. • Tuber oligospermum (Tul & Tul) Trappe Tuber pulchosporum (Polemis et al 2019)

Generalist truffle species of Balkan Peninsula – detected in most environments where the soils are not acidic, and there are ECM hosts

Tuber excavatum s.l.

Tuber rufum s.l.

Tuber aestivum Tuber brumale s.l. Tuber aestivum/ uncinatum – probably the best production in natural habitats (Serbian sandy soils with argentea, Romanian hilly regions) and plantations (Hungarian Jazsac region)

• T. aestivum helps establishment of the young ECM trees in aerated neutral soils mostly in predominantly continental climate Generalists of hilly regions – very rare in Serbia, common in Hungary

Tuber borchii Tuber puberulum Maljen mountain Specialised for forests on very alkaline soils in hills

T. fulgens T. mesentericum T. oligospermum

Loess hills in Panonian plain Gradac river gorge Jablanik mountain In special reservation mountain Tara only

T. monosporum T. petrophilum Generalists of the reparian forests

T. macrosporum T. maculatum T. foetidum Tuber magnatum specialist for specific reparian forests:

The main characteristic of white truffle habitats in Serbia : • almost always in lowland forests exposed to some kind of fluvial activity – river banks (three dimensional movements) or ground water (mostly vertical movements); • wide alluvial plains always exposed to sedimentation from calcareous upland regions; •always connected to old, stable forests (Quercus robur, sp, Carpinus betulus, Ulmus minor…) in borderline zones between Pannoninal region and Dinarides

T. magnatum helps surviving of mature ECM trees in the climatic borderline zones (continental to Mediterranean) and borderline forests in hygromorphic neutral to alkaline soils

Hungary Central Serbia

North Serbia Istra, Croatia Somewhere near Croatian coast...

T. melanosporum helps establishment of young trees in predominantly Mediterranean climates on poorly developed sceletogenous soils and high pH Some species present in neighbouring areas, but so far not officially recorded in Serbia

(Croatia)  Tuber magentipunctatum Z. Merényi, I. Nagy, Stielow & Bratek, Persoonia, 38: 381 (2017). Hungary  Tuber cryptobrumale Z. Merényi, T. Varga & Z. Bratek, Mycol Progr., 16 (6): 619 (2017) (Hungary and probably Serbia)  Tuber panniferum Tul. & C. Tul (Greece)

Probably new species of T. brumale s.l. from Serbia Apart from true truffles, app 50 of other species forming hypogeic sporocarps so far in Serbia (in Hungary much more due to the existance of people working on ).

Gyromitra sp.

Helvella sp.

Helvelaceae, Balsamia vulgaris Discinaceae, Hydnotria tulasnei Desert truffles – special kind of symbiosis ectomycorrhiza with herbaceous plants, in Serbia one species of Terfeziaceae

Terfezia arenaria Pezizaceae, Mattiralomyces terfezioides establishes very specific relation with Robinia pseudoacacia (black locust) roots (fabaceous tree that normally establishes arbuscular mycorrhiza)

Peziza sp.

The colonization of the roots by the septate hyphae of M. terfezioides was weak, particularly compared to the colonization by arbuscular–mycorrhizal fungi. Kovács, G., Jakucs, E., & Bagi, I. (2007). Identification of host plants and description of sclerotia of the truffle Mattirolomyces terfezioides. Mycological Progress, 6 1, 19-26. Pezizaceae, Pachyphloeus citrinus

Pyrenomataceae, Genea fragrans, G. sphaerica, G. vagans.G. verrucosa

Morchelaceae Fischerula macrospora

Pyrenomataceae, Stephensia bombycina Common in conifer forests and / or acidic soils

Ascomycotina, Tuberaceae Choiromyces meandriformis

Boletales, Rhizopogon sp

Ascomycotina , Elaphomycetaceae, E. aculeatus, E. anthracinus E.atropurpureus, E. granulatus, E. leveillei , E. maculatus, E. muricatus , E. papillatus

Psylocybe sp. Hymenogaster populetorum Hymenogasteraceae, Hymenogaster albus Hymenogaster bulliardii Hymenogaster griseus Hymenogaster hessei Hymenogaster luteus Hymenogaster luteus var. subfuscus Hymenogaster lycoperdineus Hymenogaster niveus Hymenogaster muticus Hymenogaster citrinus Hymenogaster vulgaris

Hymenogaster rehsteineri

Hymenogaster bulliardii Phallomycetidae, Hysterangium clathroides, H. stoloniferum

Gomphaceae, Gautieria morchelliformis Geastraceae, Schenella simplex Protected and rare Rusullales, Rusullaceae: Arcangeliella (Lactarius) stephensii

Rusulales, Albatrellaceae, Leucogaster nudus Boletales, Rhizopogon vulgaris, R luteolus, R. roseolus

Boletales, Octavianina asterosperma

Boletales, Sclerogaster gastrosporioides S. hysterangioides Melanogaster ambiguus, M. broomeanus M. tuberiformis Some practical information The reality of truffle story in Serbia - trade still in the gray zone

Total mushroom export from Serbia (data from Custom service) and allowed to collect (data from INCS)

25000000

20000000

15000000 mass (kg) value (Eur) 10000000 issued total (kg) 5000000

0 2004200520062007200820092010201120122013201420152016

Relationship between yearly quota and issued permissions for truffle species in kg 1,200 Tuber magnatum Pico quota 1,000

800 Tuber magnatum Pico issued

600 Tuber aestivum Vittad. quota

400 Tuber aestivum Vittad. issued 200 Tuber brumale Vittad. / Tuber 0 macrosoprum Vittad. quota 200520062007200820092010201120122013201420152016 Protection and conservation in Serbia do not exists…. Plantation trials Istria Serbia, T. aestivum trial T. melanosporum trials, first production in Balkan Peninsula in December 2018 The T. magnatum “plantation”

Thank you for your attention!