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Introduction to and their use – – Evolution Christina Wedén – Phylogeny Uppsala University – The concept August 2019 – False and true truffles – Sexual and asexual reproduction

What is a truffle? What is a ?

• historically part of botany • Still part of botany as a scientific subject • Eukaryotic cells (true nucleus with chromosomes) • Common ancestor with animals • Unicellular (yeasts) and multicellular (mycelial) fungi

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Carl von Linné 1707-1778 Elias Fries 1794-1878 The flower king The father of mycology

Life on Earth Eukaryotes... Phaeophyta Chrysophyta Xanthophyta EUKARYOTA Oomycota ... Fungi Dinoflagellata Ciliata Apicomplexa Animalia Rhodophyta Myxomycota

Chlorophyta Entamoeba Cyanophyta Amoeba Kinetoplastida

!-proteo Euglenophyta

Trichomonads Microsporidia EUBACTERIA Diplomonads

ARCHAEBACTERIA

Redrawn and adjusted after M. L. Sogin, Current Opinions in Genetics and Development 1: 457 (1991) by Anders Backlund Från Baldauf et al., The Deep Roots of Eukaryotes Science 300: 1703-1706 (2003).

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biology - chemistry

Yeast cells and human cells • Biosynthesis of substances in organisms, and share a major part of their cell biology. in different parts of organisms, to enable life

…Although major differences in life forms! • Primary metabolites (substances) carry out the basic functions of the cell

• Specialized metabolites (”secondary metabolites”) enable survival of the organism and of the species

Two examples of specialized metabolites The fungal cell from fungi are:

• penicillin produced by Penicillium species (moulds) to fight bacteria in competition for subtrate

• sulphur containing volatiles produced by truffle species to attract vectors

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• = fungal cell • Hypha grow linked to each • Linked hypha grow out to form a branched mycelia

• Fruiting bodies are the part of the fungus that Fungi grow as mycelia we often see, but only consists of around 1% – a 3D network of elongated of the total fungal biomass – 99% is vegetative fungal cells (hyphae) mycelia. deeplivingproject.com.au Source: Source:

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What is a truffle? • Although a single hyphal strand is not visable to the naked eye, densely growing mycelia can • Fungal species which have hypogeous be seen e.g. under bark, growing on the (subterranean) fruiting bodies kitchen compost or as mycelial tufts growing • Fruiting body = stucture carrying the out from mycorrhizal roots (shown below). which are the result of the fungus’ sexual reproduction • The fruiting body is made up of vegetative

mycelia and spore tissue Vegetative mycelia Vegetative mycelia = white ”veins” = white tissue Mature spores Mature spores = brown tissue = brown tissue

Outer peridium • The truffle life form has evolved multiple times in different groups of fungi Inner peridium

Gleba Elaphomyces granulatus, the deer truffle (hjorttryffel) • Truffles are not an evolutionary distinct group, but instead belong to diverse fungal families

Peridium • The truffle life form is thought to be an adaption to e.g. drought Gleba

Tuber mesentericum, the Bagnoli truffle and aestivum, the Burgundy truffle (bagnolitryffel) (bourgognetryffel)

5 286 Phytochem Rev (2010) 9:279–301

species of macrofungi are known to science, but vectors, and to make them unearth and eat the truffles, 286 Phytochem Rev (2010) 9:279–301 estimations show that this may well be only a tenth of thereby spreading the spores. Depending on the species of macrofungi are known to science, but vectors, and to make them unearth and eat the truffles, the actual number of all living macromycete species animal vector’s lifestyle and ecology, these spores estimations show that this may well be only a tenth of thereby spreading the spores. Depending on the the actual number of all living macromycete species animal vector’s lifestyle and ecology, these spores (Hawksworth 2001). For fungi in the widest sense, the may be deposited in the shape of fecal pellets close to (Hawksworth 2001). For fungi in the widest sense, the may be deposited in the shape of fecal pellets close to total number of species on Earth has been estimated to the roots of mycorrhizal trees (Trappe and Castellano total number of species on Earth has been estimated to the roots of mycorrhizal trees (Trappe and Castellano 2019-08-12 1.1 million (Mueller et al. 2007). Within the fungal 1991). Truffles also show high levels of endemism 1.1 million (Mueller et al. 2007). Within the fungal 1991). Truffles also show high levels of endemism kingdom lies a vast, and to a large extent, still (Mueller et al. 2007), which may be explained by unexplored potential for future pharmaceutical drugs. mammalian fecal pellets being their main mean of kingdom lies a vast, and to a large extent, still (Mueller et al. 2007), which may be explained by In the section Fungi in Linnaeus’ Materia Medica spore dispersal (Fig. 5). (1749), the is mentioned as an unexplored potential for future pharmaceutical drugs. mammalian fecal pellets being their main mean of aphrodisiac and remedy against flatulence. Most In the section Fungi in Linnaeus’ Materia Medica spore dispersal (Fig. 5). probably this passage refers to what today is known (1749), the genus Lycoperdon is mentioned as an as the deer truffle, Elaphomyces granulatus, earlier Basidiomycetes have basidiospores. The known under the name Boletus cervini. In more So called false truffles belong to the basidiomycetes recent sources the deer truffle has been reputed for its basidiumaphrodisiacis a structure which andholds remedyand against flatulence. Most abilities to induce oestrus manifestation in cows dischargeprobably the spores this. passage refers to what today is known (Klintberg 1998), forming a somewhat unsuspected link to the Linnaean indication. The edible truffles of as the deer truffle, Elaphomyces granulatus, earlier the genus Tuber have historically been used as an aphrodisiac by both men and women and in modern known under the name Boletus cervini. In more science small amounts of the steroid androstenol has recent sources the deer truffle has been reputed for its been found in mature specimens of the black truffle . Androstenol is a major com- abilities to induce oestrus manifestation in cows ponent of the boar pheromone, excreted in the saliva (Klintberg 1998), forming a somewhat unsuspected during mating (Claus et al. 1981). Truffles are an intricate fungal life form occurring link to the Linnaean indication. The edible truffles of in both the two major groups of macromycetes: asco- and basidiomycetes (see Fig. 4). Only within the Ascomycetesthe genushave ascosporesTuber . haveThe historicallyis been used as an ascomycetes, species that develop truffle fruit bodies a sporeaphrodisiacsack which encloses by boththe spores menand and women and in modern are estimated to have evolved at least fifteen times in open to discharge them at spore maturity. 286 multiple families (Læssøe and Hansen 2007). Thus, Phytochem Revscience (2010) small 9:279–301 amounts of the steroid androstenol has from a modern phylogenetic perspective, truffles are regarded as a life form rather than a specific group of been found in mature specimens of the black truffle species of macrofungifungi. Despite are this, known all truffle species, to science, regardless of but vectors, and to make them unearthTuber and eat melanosporum the truffles, . Androstenol is a major com- phylogenetic placement, live in mycorrhizal symbio- estimations showsis that with thisvascular may plants. well Due to theirbe onlyhypogeous a tenthnature of thereby spreading the spores. Dependingponent of the on boar the pheromone, excreted in the saliva and subsequent loss of active spore dispersal, all the actual numbertruffle of species all living are also dependant macromycete on an animal vector species animal vector’s lifestyle and ecology,during these mating spores (Claus et al. 1981). (Hawksworth 2001for). spore For dispersal. fungi The in evolution the widest of a hypogeous sense, fruit the may be deposited in the shape of fecalTruffles pellets closeare an to intricate fungal life form occurring body might be an adaptation to dry climatic condi- total number of speciestions, but on the Earth relationship has between been the estimated hypogeous Bohlin to et al.the 2010 roots of mycorrhizal trees (Trappein both and the Castellano two major groups of macromycetes: asco- fungus, the tree and the animal vector seem to be 1.1 million (Muellerimperative et for al. the2007 life and). distribution Within of this the life form fungalFig. 4 Phylogeny1991 of). fungi Trufflesphylogeny of asco- also and basidiomy- show high levelsand basidiomycetes of endemism (see Fig. 4). Only within the cetes with focus on taxa with hypogeous fruiting bodies, kingdom lies asuggesting vast, and a more sophisticatedto a large fungal extent,niche against stilltruffles, based(Mueller on a combination et of al. several2007 published), studies which mayascomycetes, be explained species by that develop truffle fruit bodies vertebrates. The dispersal vectors may be mice, (Læssøe and Hansen 2007; Hibbet et al. 2007; Celio et al. unexplored potentialsquirrels, for deer future or wild pharmaceuticalboars (Trappe and Castellano drugs.2006). Taxamammalian labeled in blue with • indicate fecal evolutionary pellets groups being theirare estimated main mean to have of evolved at least fifteen times in 1991). Consequently the truffles, i.e. the hypogeous that have developed truffles. The most well known truffle- In the section Fungi in Linnaeus’ Materia Medicaproducing family,spore , dispersal comprising(Fig. the edible white5). and multiple families (Læssøe and Hansen 2007). Thus, spore-producing fruiting bodies, have developed a Burgundy truffles is further emphasized in bold lettering. (1749), the genusdiverseLycoperdon set of volatile substancesis mentioned to attract these as anFigure by Christina Wede´n and Anders Backlund from a modern phylogenetic perspective, truffles are aphrodisiac and remedy against flatulence. Most So called false truffles belong to theregarded basidiomycetes as a life form rather than a specific group of False truffles 123 probably this passage refers to what today is known fungi. Despite this, all truffle species, regardless of as the deer truffle, Elaphomyces granulatus, earlier phylogenetic placement, live in mycorrhizal symbio- • ”Squischy” (”rubber ball”) known under the name Boletus cervini. In more sis with vascular plants. Due to their hypogeous nature • Often less pleasant aroma recent sources the deer truffle has been reputed for its and subsequent loss of active spore dispersal, all abilities to induce oestrus manifestation in cows truffle species are also dependant on an animal vector • Hymenogaster is one example of a genus of (Klintberg 1998), forming a somewhat unsuspected for spore dispersal. The evolution of a hypogeous fruit false truffles belonging to the order Agaricales link to the Linnaean indication. The edible truffles of body might be an adaptation to dry climatic condi- the genus Tuber have historically been used as an tions, but the relationship between the hypogeous aphrodisiac by both men and women and in modern fungus, the tree and the animal vector seem to be science small amounts of the steroid androstenol has imperative for the life and distribution of this life form Fig. 4 Phylogeny of fungi phylogeny of asco- and basidiomy- cetes with focus on taxa with hypogeous fruiting bodies, been found in mature specimens of the black truffle suggesting a more sophisticated fungal niche against truffles, based on a combination of several published studies Tuber melanosporum. Androstenol is a major com- vertebrates. The dispersal vectors may be mice, (Læssøe and Hansen 2007; Hibbet et al. 2007; Celio et al. ponent of the boar pheromone, excreted in the saliva squirrels, deer or wild boars (Trappe and Castellano 2006). Taxa labeled in blue with • indicate evolutionary groups during mating (Claus et al. 1981). 1991). Consequently the truffles, i.e. the hypogeous that have developed truffles. The most well known truffle- producing family, Tuberaceae, comprising the edible white and Truffles are an intricate fungal life form occurring spore-producing fruiting bodies, have developed a Burgundy truffles is further emphasized in bold lettering. in both the two major groups of macromycetes: asco- diverse set of volatile substances to attract these Figure by Christina WedeHymenogaster´n and Andersgriseus Backlund and basidiomycetes (see Fig. 4). Only within the ascomycetes, species that develop truffle fruit bodies are estimated to have evolved at least fifteen times in 123 multiple families (Læssøe and Hansen 2007). Thus, from a modern phylogenetic perspective, truffles are 6 regarded as a life form rather than a specific group of fungi. Despite this, all truffle species, regardless of phylogenetic placement, live in mycorrhizal symbio- sis with vascular plants. Due to their hypogeous nature and subsequent loss of active spore dispersal, all truffle species are also dependant on an animal vector for spore dispersal. The evolution of a hypogeous fruit body might be an adaptation to dry climatic condi- tions, but the relationship between the hypogeous fungus, the tree and the animal vector seem to be imperative for the life and distribution of this life form Fig. 4 Phylogeny of fungi phylogeny of asco- and basidiomy- cetes with focus on taxa with hypogeous fruiting bodies, suggesting a more sophisticated fungal niche against truffles, based on a combination of several published studies vertebrates. The dispersal vectors may be mice, (Læssøe and Hansen 2007; Hibbet et al. 2007; Celio et al. squirrels, deer or wild boars (Trappe and Castellano 2006). Taxa labeled in blue with • indicate evolutionary groups 1991). Consequently the truffles, i.e. the hypogeous that have developed truffles. The most well known truffle- producing family, Tuberaceae, comprising the edible white and spore-producing fruiting bodies, have developed a Burgundy truffles is further emphasized in bold lettering. diverse set of volatile substances to attract these Figure by Christina Wede´n and Anders Backlund

123 286 Phytochem Rev (2010) 9:279–301

species of macrofungi are known to science, but vectors, and to make them unearth and eat the truffles, estimations show that this may well be only a tenth of thereby spreading the spores. Depending on the the actual number of all living macromycete species animal vector’s lifestyle and ecology, these spores (Hawksworth 2001). For fungi in the widest sense, the may be deposited in the shape of fecal pellets close to total number of species on Earth has been estimated to the roots of mycorrhizal trees (Trappe and Castellano 286 Phytochem Rev (2010) 9:279–301 2019-08-12 1.1 million (Mueller et al. 2007). Within the fungal 1991). Truffles also show high levels of endemism kingdom lies a vast, and to a large extent, still (Mueller et al. 2007), which may be explained by REVIEWS speciesunexplored of macrofungi potential are for futureknown pharmaceutical to science, but drugs. vectors,mammalian and to make fecal them pellets unearth being and their eat mainthe truffles, mean of estimations show that this may well be only a tenth of thereby spreading the spores. Depending on the In the section FungiThe biogeographyin Linnaeus’ of some secotioidMateria fungi supports Medica this spore dispersal (Fig. 5). the actual number ofclimatic all livinginterpretation macromycete of the advantages of species hypogeous fruit- animal vector’s lifestyle and ecology, these spores (1749), the genusing. ForLycoperdon example, the large andis widespread mentioned mushroom as genus an (Hawksworthaphrodisiac2001 and).Cortinarius For remedy fungi includes in against several thewidest secotioid flatulence. forms, sense, which the are Most found may be deposited in the shape of fecal pellets close to total number of speciesin forests on Earthin Western has Australia, been Kashmir estimated and North toAmerica the roots of mycorrhizal trees (Trappe and Castellano probably this passagethat experience refers either toseasonal what drought today or freezing is known of soi14J. 1.1 million (MuellerHypogeous et al. species2007 are). a Within major part of the the flora fungal of ECMfungi 1991). Truffles also show high levels of endemism as the deer truffle,in alpineElaphomyces and subalpine regions granulatus and in forests subject, earlier to sea- kingdom lies a vast,sonal anddrought toin North a largeAmerica and extent, Australia*iJaJ. still (Mueller et al. 2007), which may be explained by known under theThe name environmentalBoletus conditions cervini thought to. favour In the more evo- unexploredrecent sources potential thelution for deer of future hypogeous truffle pharmaceutical fruiting has beenmust have reputed arisen drugs. many for times its mammalian fecal pellets being their main mean of during the periods of cooling and drying associated with Inabilities the section toFungi induceglacialin cycles oestrus Linnaeus’ in the Pleistocene. manifestationMateria These cycles Medica probably in cows also spore dispersal (Fig. 5). caused episodes of forest fragmentation, which would have T------l(1749 (Klintberg), the genus1998Lycoperdonpromoted), forming multiple aisindependent somewhat mentioned evolutions unsuspected asof hypogeous an aphrodisiaclink to the and Linnaean remedyfruiting in indication. closely against related flatulence.taxa. The edible truffles Most of probably this passageMycophagy refers by to mammals what todayand mechanisms is known of spore the genus Tuberdispersalhave historically been used as an as theaphrodisiac deer truffle, by bothElaphomycesDiversity men of mycophagous andgranulatus women mammals and and, earlier indietary modern fungi Many species of small mammals that live in forests with knownscience under small the amounts namehypogeousBoletus ECM of thefungi eat steroid cervini the fruit-bodies androstenol. In of morethese fungi. has In Eucalyptus forests in Australia, for example, this includes (4 I 7 recentbeen sources found the in deer maturemost truffleterrestrial specimens hasherbivorous been of reputedand the omnivorous black for itsmammals truffle abilities to inducebelow oestrus a body manifestationsize of about 3 kg (Fig. in2), while cows in North Tuber melanosporumAmerican. and Androstenol European forests many is aground major squirrels com- and (Klintbergponent1998 of the), boarformingmicrotine pheromone, rodents a somewhat are extensively excreted unsuspected mycophagousi. in the saliva Small mammals eat a very wide taxonomic range of hypogeous link toduring the Linnaean mating (ClausECM indication. fungi: et species al. The 1981from edible39). genera trufflesof hypogeous of fungi representing 17 families and three of the five subdivisions the genusTrufflesTuber arehave anof the intricate historically true fungifungal (Ascomycotina, been life used formBasidiomycotina asoccurring an and Zygomycotina) have so far been recorded in the diets of aphrodisiacin bothby the both two majorAustralian men and groupsmammals8. women of macromycetes: and in modern asco-

scienceand small basidiomycetes amountsSpore of survival the(see steroidand Fig. germination4). androstenol Only within has the Fig. 1. Examples of (a) epigeous, (b) subhypogeous secotioid and (c) hypogeous basidiomycete fungi. Whole fruit-bodies are shownbeen on the left,ascomycetes, found cross-sectional in views mature speciesMammals specimens that disperse develop ofthe spores the truffle blackof hypogeous fruit truffle fungi bodies in two on the right. Spore-bearing tissues are shown in dark stippling on the cross-sectional ways. First, spores may be liberated into the air when the views, sterile tissues are lightly stippled. The fungiTuber representedare melanosporumare estimatedtypical members tofruit-bodies have. Androstenol evolved are unearthed at is leastand a broken major fifteen open com-by times mammals. in of the Cortinariaceae, with the epigeous, secotioid and hypogeous forms being Second, and probably of much greater importance, many species of Cortinarius,Thaxterogaster and Hymenogaster, respectively. ponentmultiple of the familiesboar pheromone,spores (Læssøe are ingested excretedand when Hansenthe fruit-bodies in the2007 salivaare). eaten Thus, and these spores are eventually returned to the soil in faeces. duringfrom mating a modern (ClausSeveral phylogenetic et al.studies1981 have ).demonstrated perspective, the viability truffles of spores are in it could be that, if Rhizopogon is very recently derived from mammal faeces, showing in the process that the faecal pel- So called true truffles belong to the ascomycetes Suillus as the molecular evidence suggests,Trufflesregarded there are are as more an a intricate lifelets formof mycophagous fungal rather mammals lifethan form a are specific an occurring excellent group source ofof than 130 species of Rhizopogodo.in bothItfungi. is clear the from two Despite these major this,ECM groups inoculum all truffle offor macromycetes:glasshouse species, plantslrJ8. regardless Identification asco- ofof examples that the systematics of many groups of fungi con- spores in faeces is the primary means by which mycophagy taining both epigeous and hypogeousandphylogenetic basidiomycetes fungi is in need of placement,is confirmed (see Fig. in livediet4 studies). in Only mycorrhizalof mammals, within and workers symbio- the rarely revision, preferably using molecular markers. report damaged spores. ascomycetes,sis with vascular species plants. thatSporesdevelop of DueECMfungi to trufflecan their germinate hypogeous fruit even bodiesif they naturehave not Climate and the selective advantage of hypogeous passed through a mammal’s gutlOJ9,but two studies suggest fruiting are estimatedand subsequent to havethat lossevolved passing of through activeat leastthe gut sporepromotes fifteen germination. dispersal, times inClaridge all The fact that the evolution of hypogeoustruffle fruiting species has areet ~1.~7 alsocompared dependant the formation on of an ECMs animal on seedlings vector that been repeated so many times multiplesuggests that itfamilies is directed by (Læssøehad faeces andof a mycophagous Hansen mammal2007 added). Thus, to their soil, some selective advantage. Thiers*from and forTrappeii a modern spore argued dispersal. that phylogenetic with seedlings The evolution that perspective, received ofuntreated a hypogeous truffles spores of areone fruitof the hypogeous fruiting is favoured under two sets of conditions: fungi that was known to have had abundant spores in the low soil moisture, and the presenceregarded ofbody physical as mightbarriers a life to beform faeces: an rather adaptation ECMs formed than on a tothe specific first dry group climatic of group seedlings condi-of but not mushroom emergence, such as freezing of soil. In dry con- on the second. And Cork and KenagyZoobserved germination ditions, the fertile tissue of epigeousfungi. fruit-bodiestions, Despite butwould this,rap- the relationshipof all some truffle spores of species, Elaphomyces between regardlessgranufatus the hypogeousin stored of faecal idly desiccate, but in hypogeous fruit-bodies it is protected pellets of a ground squirrel, but not in an untreated sample by a firm enveloping peridium,phylogenetic whichfungus, is further protected the placement, treeof spores and live of the the insame animal mycorrhizal fungus stored vector in the symbio- seemsame way. to be by the soil. Also, the formation of manyimperative small chambers for the life and distribution of this life form Fig. 4 Phylogeny of fungi phylogeny of asco- and basidiomy- within the fruit body probablysis retains with moisture. vascular Where plants.a Nutritional Due value to theirof hypogeous hypogeous fungi, and naturespecialization cetes with focus on taxa with hypogeous fruiting bodies, layer of frost or humus presents a physicalsuggesting barrier to emer- a morefor mycophagy sophisticated fungal niche against gence of mushrooms, hypogeousand fruiting subsequent has an immediate loss Chemical of active analysis spore of fungal dispersal,fruit-bodies suggests all that truffles, based on a combination of several published studies advantage: it is easier for the fungustruffle tovertebrates. lie species below the surface are The alsothey dependant dispersalare of low nutritional vectorson anquality*O. animal may Fungal vector becell walls mice, are (Læssøe and Hansen 2007; Hibbet et al. 2007; Celio et al. and have a mammal break through the barrier than to force composed of complex carbohydrates likely to be of low di- 2006). Taxa labeled in blue with • indicate evolutionary groups its way to the surface by its ownfor growth sporesquirrels, energy. dispersal. deer The orgestibility, wild evolution and boars much ofof (Trappe their a hypogeous nitrogen and (N) Castellanois bound fruit in cell 1991). Consequently the truffles, i.e. the hypogeous that have developed truffles. The most well known truffle- 504 body might be an adaptation to dryTREE uol. climatic II, no. 12 December condi- 1996 producing family, Tuberaceae, comprising the edible white and spore-producing fruiting bodies, have developed a Melanogaster broomeianus tions, but the relationship between the hypogeous Burgundy truffles is further emphasized in bold lettering. fungus,diverse the tree set ofand volatile the animal substances vectorto seem attract to be these Figure by Christina Wede´n and Anders Backlund imperative for the life and distribution of this life form Fig. 4 Phylogeny of fungi phylogeny of asco- and basidiomy- cetes with focus on taxa with hypogeous fruiting bodies, suggesting123 a more sophisticated fungal niche against truffles, based on a combination of several published studies vertebrates. The dispersal vectors may be mice, (Læssøe and Hansen 2007; Hibbet et al. 2007; Celio et al. squirrels, deer or wild boars (Trappe and Castellano 2006). Taxa labeled in blue with • indicate evolutionary groups 1991). Consequently the truffles, i.e. the hypogeous that have developed truffles. The most well known truffle- 7 producing family, Tuberaceae, comprising the edible white and spore-producing fruiting bodies, have developed a Burgundy truffles is further emphasized in bold lettering. diverse set of volatile substances to attract these Figure by Christina Wede´n and Anders Backlund

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Elaphomyces granulatus, deer truffle Tuber melanosporum, Périgord truffle Order Family Tuberaceae

Biological systematics Taxonomic rank

• Explicit goal to explore the origin and evolution of organisms

• Achieved largely by formulating phylogenetic hypotheses

• Systematically organizing living (and extinct) organisms: Navigating evolutionary space

• Results are dependent on previous knowledge – your hypothesis and what you compare your sample data to, i.e. your reference matrix

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Tuber aestivum Tuber Tuberaceae

Pezizomycetes

Ascomycota Fungi

Canis lupus ssp. familiaris Biological species concept Canis ”The biological species concept defines a species Canidae as members of populations that actually or potentially interbreed in nature, not according Carnivora to similarity of appearance. Although Mammalia appearance is helpful in identifying species, it does not define species.” Chordata

Animalia evolution.berkeley.edu/evolibrary/

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Scientific species name Example: From a philosophical standpoint: • Binomial system (Linneaus): Consists of the name of the genus (Tuber) followed by a species epithet (aestivum) What are the main differences between what Linnaeus thought then (i.e. +250 years ago), • Scientific names accepted internationally International code of nomenclature ICN: and what we believe today? https://www.iapt-taxon.org/nomen/main.php

• Common names may differ in different languages and even in different regions and dialects.

Phylogenetics A phylogeny is created by a phylogenetic • The study of the evolutionary history and analysis of characters from the included taxa: relationships among individuals or groups of organisms – Historically: Morphology, physiology

• Monophyletic groups – all members (e.g. species) – of the group have a common ancestor Molecular biology: Efficient methods for sequencing DNA. Enormous increase in number of characters to analyse • Root of the phylogenetic tree – linking the group to the common ancestor: Where the group is connected to the rest of the tree of life – Computerized analyses of DNA data: Co-analyses of distantly related groups of organsims possible

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tolweb.org Truffle life cycle • The life form of truffles have different evolutionary origin.

• Regardless of evolutionary origin, all known truffle species are:

– Ectomycorrhizal – loss of ability to break down organic material to carbohydrates (although a few exceptions exist)

– Vector mediated spore dispersal – loss of active spore dispersal

Maturity of spores

Top: 100% spores mature and full aroma.

Middle: Parts of the gleba with mature spores, others still immature. Faint aroma.

Bottom: No mature spores and only a faint ”mushroom” aroma. Illustration source: www.tuber.it

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Known spore vectors ectomycorrhiza

• Ecto = on the outside • Mice • Squirrels • Mycor = fungus • Wild boar • Deer • Rhiza = root • Soil fauna = fungus-root structure where the fungus is growing on and in the outer surface of the root (not penetrating the • Pigs know how to find truffles, but even cows and cells in the root as opposed to endomycorrhiza) dogs are known to find truffles on their own account

• Young pine tree seedling with ectomycorrhizae

www.slideshare.net

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Symbiosis - mutual benefits

• For the fungus: Supplied with carbohydrates from the plant’s photosynthesis

• For the plant: Efficient water and nutrient uptake (P, N, minerals). Physical protection against root pathogens.

Oak root (Quercus robur) with Tuber aestivum mycorrhizae. Fewer side roots with clusters of mycorrhizae.

Reproduction in fungi Sexual reproduction • A dicaryotic mycelium (two nuclei in each • Bot sexual and asexual reproduction results in cell/hypha) undergo karyogamy and meiosis haploid spores, which under the right to form spores. All spores are different from conditions can germinate and give rise to a each other (like siblings) new mycelium • In truffles the result of sexual reproduction is the truffle fruiting bodies (’truffles’) carrying the spores

Tuber rufum, red truffle (rödbrun tryffel)

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Asexual reproduction Asexual reproduction in truffles

• No recombination of genes • For some truffle species • Genetic material of the spore identical to of the genus Tuber, parent mycelium spore mats on the soil • Typical examples in moulds surface have been such as Penicillin species observed.

• Smaller spores which

may be wind dispersed Healy et al. 2012

Evidence for exisitence of mating types and loss of ability to break down organic material Mating types in sexual reproduction found in the analyses of the total genome sequencing of the Périgord truffle, T. melanosporum

One mating type often become dominating

Asexual spores is a way to introduce the other mating type to enable sexual reproduction

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Pharmacological use of truffles

• The deer truffle, Elaphomyces granulatus (and possibly also related species) was listed in the old pharmacopea • Against flatulens and as an aphrodisiac • Traditional and ongoing use for cows (Afzelius 1787)

The Telegraph 31 Oct. 2010

Searching for steroids in extracts of Elaphomyces granulatus

Steroid hormone or pheromone which trigger steroid production?

Steroids known from other truffles – androstenol in Tuber species

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But found phytosphingosine

Sulphur rich substances from truffles Elaphomyces granulatus (deer truffle) used in skin products

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Previously isolated from Elaphomyces Truffles in gastronomy granulatus: syringic acid COX-2 inhibitor (antiinflammatory) • Desert truffles, Terfezia and Tirmania species, were eaten in ancient Egypt 2000 B.C.

• In the Roman empire around year 400, both desert truffles and Burgundy truffles (Tuber aestivum syn. T. uncinatum) were eaten

Terfezia and Tirmania spp. grow with rock roses (Cistus) and Helianthemum spp. in the desert

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shoro – Rhizopogon roseolus Delicacy in Japan

• Cultivated successfully in New Zealand • Used for its texture rather than taste • Immature fruiting bodies are used (firm and pale peridium)

Tuber species

• Aroma most important Truffle aroma penetrates • Texture the egg shell and is absorbed by the egg white and yolk. • Visual – marbled gleba, black or white peridium Scrambled eggs Mayonnaise Marengues…

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Truffled cheese (brie), ice cream and butter

Soup with poached truffled egg and truffle slices

Andreas Hedlund’s Nobel lauriate dinner

”Guinea hen mosaic, carrot variation with Gotland truffle, and chanterelle duxelle with truffle mayonnaise”

19 2019-08-12

• A food product can contain truffles, without having its aroma from those truffles

• Conserved/tinned truffles: Contains the correct species, but it may be immature and regardless of maturity it will have lost most aroma in the heating process

Oregon Truffle Festival (USA)

Oregon truffle festival: Oregon white truffle, Oregon truffle festival: Oregon black truffle, Leucangium carthusianum Familiy Hellvellaceae – the morel family (Morchella, Swe: murklor)

20 2019-08-12

Thank you for your attention!

Parasite fungus on Elaphomyces Related to the insect parasites granulatus: Elaphocordyceps sp. Cordyceps spp.

Fly infected by Cordyceps dipterigena showing fruiting bodies emerging from the fly body.

21 2019-08-12

Codyceps spp. are used in traditional medicine in Asia.

Truffles may be infected without visable signs. Possible explanation to alledged pharmacological effects of deer truffles?

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