sign in sign up
<<
Home , Albatrellus, Auriscalpium vulgare, Basidiocarp, Beenakia, Cantharellaceae, Cantharellus cibarius

http://www.jstor.org/stable/3761626 .

Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at . http://www.jstor.org/page/info/about/policies/terms.jsp

. JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact [email protected].

.

Mycological Society of America is collaborating with JSTOR to digitize, preserve and extend access to Mycologia.

http://www.jstor.org

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions Mycologia, 91(6), 1999, pp. 944-963. ? 1999 by The Mycological Society of America, Lawrence, KS 66044-8897

Phylogenetic relationships of cantharelloid and clavarioid Homobasidiomycetes based on mitochondrial and nuclear rDNA sequences

Elizabeth M. Pine1 tion, fungi, , phylogeny, ribosomal Department of and Microbial Biology, DNA, University of , Berkeley,California 94720 David S. Hibbett INTRODUCTION Michael J. Donoghue Harvard UniversityHerbaria, Fruiting bodies of cantharelloid and clavarioid and Department of Organismic Evolutionary Biology, Homobasidiomycetes include funnel-shaped or pile- Harvard Cambridge, 02138 University, ate sporocarps with smooth, wrinkled, or lamellate hymenophores, and unbranched club or branched coralloid sporocarps with smooth or folded hymeno- Abstract: data from mitochondrial and Sequence phores. Ecological habits range from saprophytism nuclear small subunit rDNA were used to estimate and parasitism to ectomycorrhizal and lichenized mu- of cantharelloid and cla- phylogenetic relationships tualisms. Anatomical and biochemical diversity is varioid diverse Homobasidiomycetes. Sixty-five found in characters such as ornamentation and were in- Homobasidiomycete investigated, reactivity, hyphal structure, patterns of meiotic divi- 23 cantharelloid and clavarioid Al- cluding species. sion, secondary compounds, and chemical structure nodes in the tree could not be though deep resolved, of pigments (TABLE I). four cantharelloid and clavarioid lineages containing Although all modern authors agree that the can- were identified. fungi (i) (Canthar- tharelloid and are polyphyletic (e.g., ellus, ) is closely related to , which 2, 4, 10, 11, 12, 14, 19, 21, 24, 43, 58, 72, 74, 83), is which is a and toothed, Stichoclavaria, simple club, evolutionary relationships of monophyletic taxa have which is coralloid. These taxa all have , not been resolved. Relatively few morphological char- stichic nuclear which is a division, synapomorphy acters have been identified that can be compared this . is supporting (ii) closely across genera, and many of these support incongru- related to and . This is relationship ent relationships. Various authors have emphasized reactions of treated supported by green sporocarps different suites of characters and consequently have with iron which is reflective of the of salts, presence proposed conflicting evolutionary histories (e.g., 10, the The nearest relatives of compound pistillarin. 12, 14 vs 19 vs 72). A preliminary phylogenetic anal- these cantharelloid and clavarioid are fungi gastero- ysis using published morphological characters failed the earth star the stink- mycetes, including , to resolve relationships among genera of cantharel- horn and the "cannon-ball , " loid and clavarioid fungi (EM unpubl). Results The clavarioid Cla- . (iii) fungi , presented here use DNA sequence data as an inde- and to be derived vulinopsis, Pterula, appear pendent and abundant source of characters for com- from the that contains most of the fun- lineage gilled parisons across diverse lineages. is related to gi. (iv) closely , This discussion treats only taxa and characters rel- which is and which is toothed, , gilled. evant to results of this study. Corner (10, 12, 14), This is united ornamenta- lineage by amyloid spore Donk (19), and Petersen (74) provide broad taxo- tion. these results that there has Although suggest nomic reviews of cantharelloid and clavarioid fungi. been extensive in mor- convergence fruiting body Selected authors' taxonomic treatments of key gen- phology, certain anatomical and biochemical features era are summarized (see TABLE II). to be appear phylogenetically informative, notably Cantharelloid and clavarioid fungi figure promi- stichic nuclear of and division, presence pistillarin, nently in hypotheses about the origin of the fleshy or of ornamentation. cyanophily amyloidity spore basidiomycetes (12, 15, 31, 32, 43, 58, 72, 83, 84). Words: evolu- Key Cantharellaceae, , Their fruiting forms can be arranged in a transfor- mation series, with simple clubs at one extreme, can- Accepted for publication June 10, 1999. tharelloid forms intermediate, and forms at 1 Email: [email protected] the other extreme. Corner (15) proposed the "Cla-

944

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions PINE ET AL: CANTHARELLOID AND CLAVARIOID PHYLOGENY 945 varia theory" of basidiomycete evolution, which Gomphaceae to the Paxillaceae" (83 p 126). Peter- treats the cantharelloid and clavarioid fungi as a sen (69) concluded that Linderomyceswas a paraphyletic group from which all other Homobasi- for , a gomphaceous whose diomycetes have been derived. Corner suggested that morphology has been described as intermediate be- the simple club with a smooth hymenophore (e.g., tween and Gomphus (81). Corner (12, Clavaria) is the ancestral state of the fleshy fungi, 15) and Petersen (71, 72) agreed with Singer that from which have been derived first club-shaped and Gloeocantharellus/Linderomycesbelongs in Gompha- cantharelloid intermediates with folded hymeno- ceae, but thought it could represent an evolutionary phores (e.g., Clavariadelphus and Cantharellus, link with Paxillaceae and some Boletaceae. whose hymenial configurations differ from true la- Anatomical features suggest that cantharelloid and mellae in the orientation of hyphae in the ), clavarioid fungi comprise several independent line- and eventually gilled . Corner's model ages (TABLEI). Spore morphology can be used to has had a strong influence on subsequent evolution- delineate three groups. Hyaline, unornamented ary hypotheses. For example, Julich (43) suggested that do not react to Meltzer's reagent or Cot- that Clavariaceae was derived from the ton Blue are characteristic of most of the known can- (jelly fungi) or their ancestors, and that Cantharel- tharelloid and clavarioid fungi. Spores with distinc- lales is the basal group of Homobasidiomycetes. Mill- tive amyloid ornamentation are found in the coral- er and Watling (58 p 439) state that "the logical ex- loid genus Clavicorona;Donk (19) used this feature, tension from the clavarioid condition among epige- along with presence of gloeocystidia, to transfer Cla- ous taxa is the cantharelloid basidiome," and suggest vicorona from Clavariaceae to . The re- that have been derived multiple times from maining spore , ochraceous with cyanophilic or- cantharelloid ancestors. Other authors agree that namentation, is found in genera with a variety of there must have been transformations among coral- fruiting body forms: Gomphus (cantharelloid), Ra- loid, cantharelloid, and agaric forms, but propose the maria (coralloid), (hydnoid), and opposite polarity, suggesting that lineages containing (resupinate), and Gloeocantharellus (= cantharelloid, coral, and club fungi have been de- Linderomyces) (agaric). Despite their macromorpho- rived from agaric ancestors (2, 30, 72, 83). logical diversity, these taxa have been grouped in The agarics Hygrocybeand Gerronema have been Gomphaceae (18, 19, 21, 44, 46, 52, 71, 83), a place- suggested as close relatives of Cantharellaceae. Hy- ment that is supported by shared green reactivity of grocybeis similar to Cantharellusin having thick, waxy, fruiting bodies treated with FeSO4. The club-shaped decurrent gills, bright orange and yellow pigments, genus Clavariadelphusalso reacts with ferric salts, but long and narrow basidia, and hyaline, unornament- has smooth, hyaline, unornamented spores (56). If ed, non-reactive spores (34). Gerronema(= Omphal- macrochemical reactions and mode of nuclear divi- ina) chrysophyllum resembles members of Cantha- sion are emphasized, Clavariadelphus is placed with rellaceae in spore color, hymenial anatomy, basidio- Gomphaceae (2, 19, 30, 32, 56, 72, 90), but emphasis carp color, general aspect, and molecular structure on spore morphology supports a relationship with of carotenoid pigments (2, 29, 48, 83). Yet chante- Cantharellaceae or Clavaria (10 p 25, 11, 12, 14, 58, relles depart from true mushrooms in several impor- 65, 68, 72). tant characters (TABLE I), including anatomical dif- The position and orientation of the first nuclear ferences between cantharelloid gill-folds and true division of has been proposed as a taxonom- agaric gills (12 p 19), leading several authors to as- ically important character (19, 41, 42, 55). In most cribe similarities between Cantharellaceae and Hygro- Homobasidiomycetes that have been examined, di- cybeor Gerronemato convergent evolution (12, 19 p vision takes place near the apex of the with 245, 33). the meiotic spindle transverse to the long axis of the Singer (83 p 126) suggested that "A further basidium (6 p 267, 39, 42, 55, 63). This pattern is 'bridge' between and called chiastic division (see FIG. 3). In contrast, in might be seen in Linderomyces,"a genus with a bilat- Cantharellus (39, 42, 55), Craterellus (42, 55), Cla- eral trama (true gills) and unusual "cocsinoid" vulina (42, 55, 63), Stichoclavaria (= ) (sieve-like) hyphae (82), but with microscopic fea- (39, 42), (6 p 267), (6 p 267, tures and chemical reactions characteristic of Gom- 49), and Hydnum s. s. (55, 63, 80), division is near phaceae (13, 69, 71). Singer originally placed Linde- the middle of the basidium, with the spindle axis romycesin Paxillaceae (82); he later concluded that more or less parallel to the basidial axis. This pattern the genus represented an independent origin of gills is called stichic division (see FIG.3). Meiotic division within Gomphaceae, but thought it might be "a start- can be observed only in fresh, mature fruiting bod- ing point for an evolution which would lead from the ies, and has not been examined in many taxa.

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions TABLE I. Selected characters of cantharelloid and clavarioid fungi and putative relatives included in this studya

Spores Basidio- carp Discon- Configura- Ornamen- Pigmen- morphologyb tinuousc tiond Thickening' Habit' tationg tationh Reactioni NumberJ - Cantharellus pi/ca (12) + (19) wr/la (12) + (12) em (8) (11, 12) hy/(pi, ye) (12) (11, 12, 65) 2-8 (11, 12) Clavaria cl/(co) (14) -(10) sm +/- (10) te (10) -/(+) (14, 72) hy/(pi') (10) -(46, 65) 4 (10) - Clavanradelphus cl/ca (56) /(+) (56) sm/wr (56) + (56) em -(56) hy/(bu) (56) (56, 65) 4/(1-3) (56)

4 Clavicoronapyxidata co (22) + (10, 22) sm -(10) wo (19) -(22) hy (22) am (19,51) (22) Clavulina cl/co (10) /(+) (10) sm + (10) em (8) - (10) hy (10) - (46) 2/(1) (10)

- 2-4 Clavulinopsisfusiformiss cl/co (78) - (10) sm (72) + (68) te (68) - (68) hy (67) (46, 65) (68) Craterellus ca (12) + (11) sm/wr (11) +/(-) (12) em -(11) hy (10) - (11) 2-6 (12) Gerronema pi (83) + la (83) - (11) li/sa (83) - (83) hy/or (83) - (83) 4 (83) Gloeocantharellus pi (71) + (13) la (71) -/(+) (13) te (12) wa (71, 81) oc (70, 81) cy (67, 70) 2-4 (13) Gomphus ca (12) + (56) sm/wr (56) + (13, 70) em (8) wa/ri (71) oc (56) cy (67) 2-4/(8) (12) - 2-6 Hydnum pi (19) + to (19) em - (19) hy (19) (19) (19) 2 or 4 pi (83) + la (83) -(11) em/sa (83) -(83) hy (34) (83) (83) 0 s.s. co +/- sm + (10) sa (67) - (19) hy (19, 67) 2-8 (10) (67) (10) 0 4 Macrotyphula" cl (3, 73) - (10) sm (10) + (56) wo (10) - (10) hy (10, 72) (3, 10) > Pterula cl/co (10) + (10) sm + (10) sa (10) - (10) hy (10) 2-4 (10) Ramaria co (10) + (10) sm +/- (10) em (8)/sa (10) ro/sp (74)/(-) (14) oc (72) cy (46, 65, 70) 1-4 (10) Stichoclavariav cl (67) - (10) sm - (67)/(+) (10) li/(sa) (67) -(67) hy (67) 2-4 (67) 2-8 Typhula subg. Typhula cl (3) + (3) sm - (10, 56) sa/pa (10) - (10) hy (10) - (3) (10) a Scoring is for the entire listed, with rare states indicated in parentheses. b = = Fruiting body morphology: cl = clavarioid (simple lub), co coralloid (branching cylinder), ca = cantharelloid (funnel-shaped), pi pileate/agaricoid. ' = Discontinuous hymenium: - = fertile area continuous across top of , + apex of fruiting body sterile (assumed true for "pi" fruiting bodies). d to la = r Hymenial configuration: sm = smooth (assumed true for clavarioid and coralloid taxa unless otherwise reported), wr = wrinkled folded, lamellate/appearing gilled, to = toothed/hydnoid. = mature basidia. Thickening hymenium (sensu 10):- = hymenial layer constant thickness throughout development, + developing basidia push past = = sa = te1 Habit/ecologial strategy (substrate reported only if specific ecological data not available): em ectomycorrhizal, li = lichenized, pa parasitic, saprobic, =terrestrial, wo = found on dead plant matter. = = g Spore ornamentation: - = none (spores smooth), ro = roughened, wa warty, ri ridged or with anastomosed warts, sp = spiny/echinulate. h = = = ochraceous. Spore pigmentation: hy = hyaline (unpigmented), pi = pink, ye yellow, or orange, bu = light , oc = = iReactivity of spore wall ornamentation (not including ): - = not blue in Meltzer's reagent or Cotton Blue, am amyloid, cy cyanophilic blank = reactivity not reported for either reagent. i Number of spores (or sterigmata) per basidium. k = Presence/type of cystidia in hymenium: - = no cystidia of any type, le = leptocystidia/undifferentiated cystidia, gl gloeocystidia (also known as cocsinocystidia)

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions TABLE I. Extended

Hyphae Biochemistry Cystidiak Skeletal' plerousm Clamps" Meiotic type? Fe reactivityP Carotenoidsq

Cantharellus - (11, 12) - (12, 65) - (11, 65) + (11)/(-) (13) st (39, 42, 55) - (19)/(lg/re [72]) + (30) Clavaria - (10) - (65) - (65) - (14, 72) ch (42, 63) vi (89)/- (72) +r/_ (30) Clavariadelphus le (56) - (10, 65) + (56, 73) + (56) ch (63: Clavaria pistillaris, C. ligulus, gr (19, 56), +pi (78) - (29 in 30, 72) C. truncatus) 3 Clavicorona pyxidata le (10)/gl (22, 23) - (10) - (10) + (22) - (22) Clavulina -/(le) (10) - (10) - +/(-) (10) st (42: Clavaria cinerea, C. cristata, 55: C. rugosa, C. grisea, 63) ,I. Clavulinopsisfusiformiso - (68) - (67) - (65) + (67, 68) ch (39, 42: Clavaria muscoides, C. subtilis, 63) gr, -pi (78) - (30, 78) Craterellus - (11) - (11) - (10) - (11) st (42: , 55) - (19) + (30) Gerronema -/(le) (83) - (83) - +/- (83) ch (Kiihner in 39, 48) + (29) H x Gloeocantharellus gl (13, 81) - (83) + (67, 70) + (71, 81) gr (71) - (71) Gomphus - (12, 70)/(le) (70) - (12) + (12, 71) +/(-) (12, 71) ch (42: Craterellus clavatus) gr, +pi (78) - (30, 71) Hydnum - (19) -(19) -+ (19) st (42, 63, 80) r - ch g Hygrophoraceae - (83) - (83) +/- (34, 83) (55) 5 Lentaria s.s.t - (10) + (65) - + (72) ch (42: Clavaria epichnoa) gr (19) Macrotyphulau - (10) - (10) la (56, 73) + (56) - (56) Pterula -/le (10) + (10) - + (10) Ramaria - (10) +/- (16, 65) + (65) +/(-) (72) ch (42: Clavaria abietina, C. crispula, 63) gr, +pi (78) - (30, 77) z7rI o Stichoclavariav -/(le) (67) -(10) - +/(-) (67) st (39, 42: Clavaria falcata) 0 Typhula subg. Typhula - (10) - (10) - +/- (10) ch (63) 0-

Skeletal hyphae in basidiocarp: - = absent (monomitic), + = present (dimitic). - Gloeoplerous hyphae in basidiocarp:- = absent (or not reported), + = present, la = laticiferous hyphae (superficially similar to gloeoplerous hyphae, but no ~t staining in Cotton Blue). 0 n Clamp connections throughout basidiocarp: - = absent (or a single clamp at the base of the basidium, as seen in Clavaria), + = present. Orientation of first meiotic nuclear division (original reports only): st = stichic (spindle oblique or parallel to long axis of basidium), ch = chiastic (spindl transverse), blank = not reported, original classification of examined specimen is listed if it was later transferred to a different genus. P Reaction of fruiting body with iron salts (i.e., FeSO4 or FeCl3): - = no reaction, lg = light green, re = red, gr = dark green, vi = violet, +pi = compounc ii pistillarin demonstrated to be present, -pi = pistillarin assayed but not present, blank = reactivity not reported. q Carotenoid pigments in fruiting body extractions: - = absent, + = present. r Characteristic of Clavaria helicoides, whose generic position is uncertain (see 14: p. 34-35). s C. fusiformis has been placed variously in , , and Clavaria, so scoring is limited to the single species. t Lentaria s.s. refers to taxa remaining after the segregation of Multiclavula Petersen. Macrotyphula scoring is based largely on Corner's (10) Clavariadelphus subg. Typhulopsis; Methven (56) equates the two. Stichoclavaria Ulbrich is used for Multiclavula Petersen, see text.

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions 948 MYCOLOGIA

Shared possession of stichic division suggests that genus (e.g., 50), or to segregate Craterellusinto its Cantharellaceae is closely related to Hydnum (17, 19, own (e.g., 43), such changes in taxonomic 72, 74) and Clavulina (17, 19). If stichobasidia are rank have not clarified relationships among cantha- deemphasized as a taxonomic character, other fea- relloid lineages. tures suggest different relationships (TABLE I). For Clavarioid basidiomycetes are a heterogeneous example, Corner (11) used hymenophore configu- group whose phylogenetic relationships have also ration, fruiting body development, clamp connec- proved extremely difficult to resolve. A few genera, tions, and presence/absence of a sterile apex or pi- such as Clavicorona and Ramaria, share distinctive leus to split Cantharellaceae, suggesting placement of features with other lineages of Homobasidiomycetes Cantharellus with Clavariadelphus, Craterellus with and have been removed from Clavariaceae (18, 19). Stereum,and Clavulina with Clavaria and Clavulinop- Other species have autapomorphic features that have sis. Petersen (66) agreed that stichic Clavulina was allowed segregation of the umbrella Clavaria into dis- related to chiastic Clavulinopsis, but placed Clava- tinct genera. For example, Clavulina is characterized riadelphus with this complex rather than with Can- by secondarily-septate basidia with two strongly in- tharellus. Reijnders and Stalpers (79) found a differ- curved sterigmata, Pterula by a dimitic hyphal system, ent pattern of hymenophore trama development in and Typhula by the formation of sclerotia (10, 14). than in Cantharellaceae, which, But such characters do not suggest higher-level rela- combined with the absence of carotenoid pigments tionships, and although some authors have promoted in Hydnum, led them to reject a close relationship these genera to segregate families (19, 43), their between Cantharellaceae and Hydnum. nearest relatives have not been identified. Stichic di- Circumscription of genera within Cantharellaceae vision (found in Stichoclavaria and Clavulina) and has been controversial (74). Craterellushas been dis- carotenoid pigmentation (found in Clavaria subg. tinguished mainly by absence of clamp connections Clavulinopsis sensu Petersen, 77), link some genera (4, 11, 12), but Petersen (74) noted that some species to other lineages of Homobasidiomycetes (TABLEI), that lack clamps have been included in Cantharellus. but these characters have not been widely accepted Corner (11) proposed the genus Pseudocraterellusto as synapomorphies. Thus Clavariaceae is still a poly- contain unclamped, secondarily septate phyletic group that is defined largely by the absence otherwise similar to Cantharellus; Corner also em- of distinguishing features. phasized patterns of fruiting body development, but this feature is difficult to examine and has been MATERIAILSAND METHODS largely ignored by subsequent workers. Petersen (70, 74) and Bigelow (4) criticized secondary septation as Twenty-fivecantharelloid and clavarioidexemplars were se- a taxonomic character since it is variable among in- lected to represent 23 species in 12 genera and 8 families dividual fruiting bodies, especially those of different sensu Corner (12, 14). Taxawere chosen to emphasizetax- ages, and is difficult to ascertain in herbarium ma- onomicallycontroversial traits (e.g., stichicnuclear division, with an effort to terial. Furthermore, many authors have noted can- spore ornamentation,FeSO4 reactivity), include of each of chanterellesand tharelloid species that exhibit combinations of fea- multiple species genus tures used to define different Craterellus several clavarioid genera (TABLEII). Sequences for Clavi- genera (e.g., corona had been or whose Corner's criteria pyxidata publishedpreviously (35). carolinensis) placements by Because of can- conflict with those other higher-level evolutionary relationships supported by well-accepted tharelloid and clavarioid fungi are controversial, a broad characters Corner himself (4, 11, 13, 70, 75). (13) sampling of other Homobasidiomycetes was imperative. pointed out that Cantharellus inathinus and C. sub- Four taxa were chosen to represent proposed relativesof cibarius can be clamped and secondarily septate; his Cantharellaceae: Hydnum repandum, Gerronemachrysophyl- description of C. cuticulatus, which is "so very obvi- lum, and two species of Hygrophoraceae. Gloeocantharellus ously a Cantharellus" (p 786) led him to conclude purpurascens, with true gills, was the sole representative of that "secondarily septate hyphae without clamps, noncantharelloid or clavarioid Gomphaceae. Sequences for such as characterize ,occur in this additional taxa were available from published and ongoing studies of (35, 36, 37). species of Cantharellus" (p 785). Despite examina- Homobasidiomyceterelationships Thirty-sixexemplars were selected to represent traditional tion of pigment structure (30), spore wall anatomy families of basidiomycetesas well as unclassifiedlineages (45), secondary septation, fruiting body ontogeny, identified by previous phylogenetic analyses.In total, 21 and (11, 12, 13), no hyphal anatomy synapomorphies families sensu Donk (19) and Singer (83) were represented. have been that distin- recognized unambiguously DNA was isolated from dried or fresh fruiting bodies or guish Craterellus, Cantharellus, and Pseudocraterellus. mycelia. Some taxa proved extremely difficult to extract, Although these difficulties have led some authors to particularly those with dark pigments (e.g., Craterellusfal- collapse all the species of Cantharellaceae into one lax), and protocols were modified to reduce the concentra-

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions TABLE II. Taxa examined, with selected authors' familial placements GenBank Taxon Collectiona Herbarium Donk (19) Corner (12. 14) JuAlich(43) nuc-ssu-rDNA mt-ssu-rDNA DSH94-006 F Cantharellaceae Cantharellaceae Cantharellaceae AF184176 AF185966 Cantharellus lutescens DA0M199243 DAOM Cantharellaceae Cantharellaceae Cantharellaceae AF184177 AF185967 Cantharellus B&COO1 F Cantharellaceae Cantharellaceae Cantharellaceae AF184178 AF185968 sp. It Cantharellus tubaeformis DSH93-209 F Cantharellaceae Cantharellaceae Cantharellaceae AF026636 AF026678 Clavaria acuta HS3785 SFSU Clavariaceae Clavariaceae Clavariaceae AF184180 AF185969 0-4 DED3622 SFSU Clavariaceae Clavariaceae Clavariaceae AF184181 AF185970 Wiejek 41 wrTU AF184182 AF185971 Clavariadelphus ligulus 21648 F Clavariaceae Clavariadelphaceae AF184183 AF185972 Clavariadelphus pistillarus 37813 TENN Clavariaceae Clavariadelphaceae Clavariadelphaceae AF026639 AF026681 t7l Clavariadelphus unicolor 36248 TENN Clavariaceae Clavariadelphaceae Clavariadelphaceae AF184185 AF185973 JFA10798 WVTU Clavulinaceae Clavulinaceae AF184186 AF185974 DA0M159321 DAOM Clavulinaceae Clavulinaceae Clavulinaceae AF026640 AF026682 2 Clavulina ornatipes DED2869 SFSU Clavulinaceae Clavulinaceae Clavulinaceae AF184188 0 EP96-004 F Clavariaceae Clavariaceae Clavariaceae AF184189 AF185975 Craterelluscornucopioides Danell43b Cantharellaceae Cantharellaceae Craterellaceae AF184190 AF185976 Craterellusfallax DSH96-003 F Cantharellaceae Cantharellaceae Cantharellaceae AF184191 AF185977 t:j Gerronemachrysophyllum RHP42170 F Tricholomataceae AF184193 AF185978 AF184194 AF185979 Gloeocantharelluspurpurascens 51702 TENN Gomphaceae Gomphaceae Gomphaceae 0n Gomphus bonarii REH1982 F Gomphaceae Gomphaceae Gomphaceae AF184195 AF185980 Gomphusfloccosus DSH94-002 F Gomphaceae Gomphaceae Gomphaceae AF026637 AF026679 0 Hydnum repandum EP96-001 F Hydnaceae AF026641 AF026683 zt Hygrocybeconica HDT48309 SFSU Hygrophoraceae AF1841 98 AF185981 HDT54064 SFSU Hygrophoraceae AF184199 AF185982 Lentaria byssiseda *HDT5502 TENN Gomphaceae Ramariaceae AF184200 AF185983 t"3 Macrotyphula cf juncea *DM-975 MIN. Clavariaceae Clavariadelphaceae Clavariadelphaceae AF184201 AF185984 Multiclavula mucidau DSH96-058 F Clavariaceae Clavariaceae Clavariaceae AF026613 AF026659 Pterula aff epiphylloides DM-937 MIN. Clavariaceae Pterulaceae Pterulaceae AF184204 AF185985 HS1788 TENN Gomphaceae Ramariaceae Ramariaceae AF184205 AF185986 *HDT5474 TENN Gomphaceae Ramariaceae Ramariaceae AF026638 AF026680 Typhula phacorrhiza DSH96-059 F Clavariaceae Clavariadelphaceae AF026630 AF026686, AF026687

a Accessions are dried fruiting bodies, except those marked with *, which are mycelial cultures. 11 Accession is a yellow mutant of the species (1, 64). "Also known as Stichoclavaria mucida.

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions 950 MYCOLOGIA tions of these pigments. Fragments of fruiting bodies were positioning of gaps was particularly ambiguous (128 bp). first soaked in a buffer of 20% DMSO, 250 mM EDTA, and Dataset 3, the most exclusive set, additionally omitted an saturated NaCl (S. Rehner pers comm) 1-3 d, then rinsed extremely variable region of mt block 5 (106 bp), and all with 1X TE pH 8.0 (10 mM Tris-HCl, 1 mM EDTA) 10 min. characters that were scored as missing for more than 10% Extraction protocol was as follows: a small fragment (0.25 of the taxa. cm3 or less) of fungal tissue was placed in a 1.5 mL micro- Dataset 2, the intermediate exclusion set, was used to an- centrifuge tube with 400 FL hot (60 C) 1% SDS extraction alyze sequences for the two genes separately. Analyses of the buffer and sterile sand. Tube contents were homogenized mitochondrial gene alone excluded Clavulina ornatipes. with a plastic pestle fitted into a hand drill (recalcitrant tissue Analyses were performed on the combined data from both was ground in a mortar under liquid nitrogen, then added genes using all three datasets. to hot buffer). Tubes were incubated at 60 C 10-30 min, After two well-supported (designated "gomphoid- then extracted once with 25:24:1 phenol: chloroform: isoa- phalloid" and "stichic") were identified in analyses includ- myl alcohol and once with 24:1 chloroform:isoamyl alco- ing all taxa, two new alignments were constructed that in- hol. DNA was removed from solution using Geneclean II cluded only members of each clade. This reduced the total (Bio 101, LaJolla, California) and eluted into 50 FL 1 X TE number of gaps required for alignment, and allowed inclu- pH 8.0 (10 mM Tris-HC1,1 mM EDTA). Serial dilutions of sion of additional characters from regions that were too genomic DNA (1:10-1:1000) were used as template for the divergent to be aligned across the complete taxon set (see polymerase chain reaction. TABLE III). Two unlinked genes were examined: mitochondrial small Phylogenetic analyses were conducted using PAUP 3.1.1 subunit rDNA (mt-ssu-rDNA) and nuclear small subunit (86) and test version 4.0d54 of PAUP* (written by David L. rDNA (nuc-ssu-rDNA). Amplification and sequencing used Swofford) on a Power Macintosh 8500/220 and Sun work- the primers MS1 and MS2 (91) for mt-ssu-rDNA, and SRlc station. Heuristic searches were performed, with 100 ran- and NS41 (35) for nuc-ssu-rDNA. Double-stranded PCR dom stepwise addition replicates with MULPARSon, steep- products were purified using Geneclean II (Bio 101, LaJol- est descent off, and TBR branch swapping. A two-step la, California) or QIAquick spin columns (QIAGEN, Inc., search was performed: first, no more than two trees were Chatsworth, California). PCR and sequencing parameters saved from each replicate, then exhaustive swapping was were as described by Hibbett and Donoghue (36). Sequenc- performed on all of the most parsimonious trees discov- es were edited and assembled using SeqEd v. 3.0.1 (Applied ered. The resulting trees were rooted with ,as sug- Biosystems, Inc., Foster City, California) or Sequencher v. gested by the results of Swann and Taylor (85). One thou- 3.0 (Gene Codes Corp., Ann Arbor, Michigan). sand bootstrap replicates were performed with the follow- Sequences were aligned manually in SeqApp v. 1.9a169 ing settings: MULPARS option off, simple addition se- and PAUP v. 3.1.1 (86); automated alignment algorithms quence, heuristic search, and TBR branch swapping. were ineffective due to extensive length variation. For the Analyses of the two subset alignments (gomphoid-phalloid number of nucleotides sequenced for each gene fragment, and stichic) used the branch-and-bound search algorithm, the size of the data matrix after introduction of alignment which guarantees discovery of all most parsimonious trees. gaps, and the number of potentially phylogenetically infor- mative characters included in analyses, see TABLE III. The RESULTS mt-ssu-rDNA alignment was divided into seven sections fol- Hibbett and blocks 1, 3, 5, and 7 lowing Donoghue (36): The number of included, variable, and parsimony- were aligned across all taxa, but blocks 2, 4, and 6 exhibited informative characters for each data set is shown in extreme variability and were excluded. Certain regions TABLE III, along with the number and length of op- could not be aligned for divergent individuals and were timal trees found in each analysis. Independent anal- scored as missing data for those taxa (mt block 1: 43 bp of of mt-ssu-rDNA and nuc-ssu-rDNA that both Clavaria zollingeri isolates; mt block 7: 53 bp of Spa- yses suggest there is rate line- rassis spathulata; nuc: 237 bp of Cantharellus cibarius and evolutionary heterogeneity among 122 bp of remaining species of Cantharellus and Craterel- ages in both genes. In the mt-rDNA tree (FIG. 1), lus). One hundred and fifty-three bp of the nuc-ssu-rDNA there are long branches leading to Clavaria zollingeri were not comparable across all taxa but could be aligned (33 steps), Sparassis spathulata (49 steps), the branch within subsets; corresponding positions in the remaining linking these three isolates (53 steps), and the taxa were scored as data. Clavulina was missing ornatipes branch linking these taxa to Stichoclavaria (34 steps). not for the mt-ssu-rDNA and was scored as miss- sequenced These are four of the five longest branches in the ing for all mt-rDNA in combined positions analyses. Align- tree; the fifth consists of the 45 autapomorphic ments are deposited in TreeBASE. changes leading to Boletus satanas. In the nuc-rDNA Three data sets were developed to explore sensitivity of tree (FIG. 2), 63 autapomorphic changes lead to Can- results to inclusion or exclusion of ambiguously aligned re- tharellus cibarius, and there is a branch of 38 gions (see TABLEIII). Dataset 1, the most inclusive, omitted long of Cantharellaceae. The only the beginnings and ends of sequences (124 bp), the steps supporting unalignable mt-ssu-rDNAblocks 2, 4, and 6, and sites scored next longest branch is 35 steps, leading to Dacrymyces as missing for all but a single isolate. Dataset 2, the inter- chrysospermus, at the base of tree; no other branch is mediate exclusion set, further excluded regions where the more than 25 steps long. The most obvious conflict

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions PINE ET AL: CANTHARELLOIDAND CIAVARIOIDPHYLOGENY 951

TABLEIII. Descriptionof the variousdata sets analyzedand the most parsimonioustrees found Characters ShortestTrees Size of Size of Data set Matrix Included Variable Informative Number Length Inclusivea 4383 1084 634 424 56 2629 Intermediatea 4383 956 522 346 4458 2022 Exclusivea 4383 744 430 283 64 1469 Mt-ssu-rDNAab 3303 311 242 183 372 1234 Nuc-ssu-rDNAa 1079 645 290 163 >4800 682 Gomphoid-phalloidc 2357 898 202 90 1 332 Stichicd 1783 1127 376 169 2 419 a Single alignment including all 65 taxa. h Clavulinaornatipes was excluded. New alignment including only the 12 taxa in the gomphoid-phalloidclade. d New alignment including only the 11 taxa in the stichic clade. between the two gene phylogenies concerns relation- monophyly of stichic taxa. Lack of resolution in the ships of taxa on these long branches. The mt-rDNA strict consensus tree is due to conflicting placements tree (FIG. 1) depicts monophyly of Clavaria zollingeri of Clavaria zollingeri and Sparassis; alternate equally and Sparassis spathulata and places these taxa as the parsimonious positions are marked with dashed sister group of Stichoclavaria, although with less than branches in FIGS. 3, 4. When Clavaria zollingeri and 70% bootstrap support. The nuc-rDNA tree (FIG.2), Sparassis were excluded from analyses, monophyly of in which these taxa are not associated with unusually stichic taxa was supported by all most parsimonious long branches, supports monophyly of all Clavaria trees and 100% of bootstrap replicates. species and Clavulinopsis, and places Sparassis as the Cantharelloid and clavarioid fungi appear in four sister group of Laetiporus. The mt-rl)NA tree (FIG.1) groups (FIG. 3). Gomphus, Ramaria, Gloeocantharel- gives strong support (99% bootstrap) for the mono- lus, Lentaria, and Clavariadelphus form a clade in- phyly of Cantharellaceae and Hydnum, but the nuc- cluding Pseudocolus, Geastrum, and Sphaerobolus rDNA tree (FIG.2) places Cantharellusand Craterellus (henceforth referred to as the gomphoid-phalloid near the base of the phylogeny, and leaves Hydnum clade), with 100% bootstrap support. The stichic gen- with the remainder of the stichic clade. Other nodes era Cantharellus, Craterellus,Hydnum, Clavulina, and that differ between the two gene trees either collapse Stichoclavaria are monophyletic, including Sparassis in the strict consensus of equally parsimonious trees and Clavaria zollingeri in some of the most parsimo- or receive less than 60% bootstrap support from one nious trees. Clavicorona is the sister group of Auris- or both genes. calpium and Lentinellus. The remaining clavarioid Results of analyses of the three exclusion sets of fungi are nested within the clade including most of the combined data (datasets 1-3) differed slightly in the gilled fungi and the Fistulina hepatica bootstrap values and degree of resolution of the strict (henceforth termed the euagaric clade after Hibbett consensus tree, but no conflicting nodes received et al 37). even moderate (>50%) bootstrap support. Because Restricting attention to members of each of the the major conclusions of this study are congruent first two clades (gomphoid-phalloid and stichic) al- with all three sets of analyses, only results of dataset lowed unambiguous alignment of more of the se- 2 will be presented. Combined analyses (FIGS.3, 4) quence data. Compared to the alignment including place Clavaria zollingeri and Sparassis together and all 64 taxa, fewer gaps were required, reducing the support monophyly of Cantharellaceae and Hydnum, matrix length, and reduced homoplasy provided few- reflecting the mt-rDNA results (FIG. 1). The branch er, shorter most parsimonious trees with better-re- leading to Clavaria zollingeri and Sparassis is the lon- solved fine-scale relationships (TABLE III). The rela- gest in the tree (68 steps). Furthermore, two of the tionships supported by the single most parsimonious three next longest branches lead to Sparassis itself tree for the gomphoid-phalloid clade realignment (60 steps) and Clavaria zollingeri (63 steps). The re- (FIG.5) are congruent with those supported by some maining unusually long branch leads to the divergent of the most parsimonious trees for analyses including Cantharellus cibarius (64 steps). The strict consensus all taxa (e.g., FIG.3). Gomphusis monophyletic (99% tree (FIG. 4) does not resolve relationships of stichic bootstrap), and closely related to Ramaria formosa. taxa, but 71% of the bootstrap replicates support Although Ramaria stricta is the sister group of this

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions 952 MYCOLOGIA

58 1 Sparassisspathulata 62u FI r Clavarizollingeri c 12lavaria zollingeri Stichoclavariamucida r Clavulina cinerea 1Clavulina cristata - Cantharelluscibarius Cantharelus lutescens stichic Cantharelussp. Cantharelustubaeformis Craterelluscornucopioides

I Pterulaffeipophylloides ._98{- Lentinulaitentia euagaric Gerronema chrysophyllum -- tuberregium Fomitopsispinicola '- Clavulinopsisfusiformis Meripilusgiganteus Bondarzewiaberkleyi Echinodontiumtinctorium 64 rStereum hirsutum 99 Aleurodiscus botryosus *r*-e lGloeocystidiellumleucoxantha \ ,54- Hygrocybe_Hygrocybe coClavaaacutaconica euagaricgarc

Auriscalpiumvulgare - Clavtcoronapyxidata - Laxitextumbicolor - Heterobasidionannosum - Russula compacta 20 steps Boletussatanas Albatrellussvringae

gomphoid- *I* Ramaia stricta phalloid - 51 Gloeocantharelluspurpurascens * Sphaerobolusstellatus J G'-(Geastrumsaccatum 100 L Pseudocolusfusiformis Schizoporaparadoxa C-- Oxyporussp. oltricia perenis Hyphodontiaalutaria - Tremellasp. Phellinus igniarius = FIG. . Phylogram of mt-ssu rDNA gene tree. One of 372 equally parsimonious trees. L 1234, CI = 0.332, RI eq 0.583, RC = 0.194. The following conventions are used in all figures: cantharelloid and clavarioid taxa are in boldface, bootstrap values (greater than 50% in FIGS.1, 2, 4-6) are indicated next to the appropriate branch, branches receiving >70% bootstrap support are thickened, * indicates branches that collapse in the strict consensus of all most parsimonious trees.

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions PINE ET AL: CANTHARELLOID AND CLAVARIOID PHYLOGENY 953

Dacrymyces chrysospermus Tremellasp. - Auriculariaauricula C Echinodontiumtinctorium * r Lentinellusomphalodes {--- Auriscalpiumvulgare _ I Clavicoronapyxidata Russula compacta Meripilusgiganteus - Phanerochaetechrysosporium *Clavariadelphuspistillaris tClavariadelphus unicolor Clavariadelphusligulus 99 Lentaria byssiseda ,.* - - 58 Gloeocantharelluspurpurascens Pseudocolusfusiformis gomphoid- * Gomphus bonarii Gomphusfloccosus phalloid 5* Sphaerobolusstellatus Geastrumsaccatum - - Ramaria stricta * L _ Ramariaformosa 91- Clavulina cinerea - - Clavulina cristata . Clavulinat ornatipes stichic 98 Stichoclavaria mucida Hydnumrepandum - Fomitopsispinicola Panus rudis - Bjerkanderaadusta Hyphodontiaalutaria _- Trametessuaveolens *- Albatrellussyringae Laxitextum bicolor r Bondarzewiaberkleyi * Heterobasidionannosum - Stereumhirsutum - Gloeocystidiellumleucoxantha Aleurodiscusbotryosus r Pleurotus tuberregium -_ Gerronemachrysophyllum Lentinulalateritia 52 Clavaria acuta 63 100 Clavariazollingeri * Clavariazollingeri Clavulinopsisfusiformis euagaric - Hygrophoruseburneus - -- - 8IMacrotyphula cf. juncea Typhulaphacorhiza - Pterula aff. epiphylloides Hygrocybeconica . 3Sparassisspathulata - r---4-_ 'Laetiporus sulphureus Botryobasidiumsubcoronatum Boletus satanas Oxyporussp. r- Peniophora nuda 1- Fistulina hepaticad euagaric - Phellinus igniarius ~53 ~ Coltriciaperenis Schizoporaparadoxa uanmtareuus!...l cioartus ... Q~99 ~66, Cantharelluslutescens 55[ Craterellusfallax stichic 100 Cantharellus tubaeformis Cantharellus sp. Craterelluscornucopioides

FIG. 2. Phylogram of nuc-ssu rDNA gene tree. One of >4800 equally parsimonious trees. L = 682, CI = 0.526, RI = 0.732, RC = 0.385. See FIG. 1 for explanation of conventions.

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions 954 MYCOLOGIA

100 Dacrymyceschrysospermus Auriculariaauricula *. Echinodontiumtinctorium _ Peniophora nuda .- Boletus satanas -:- 57r Stereumhirsutum " Aleurodiscus 100 botryosus 20 steps -- Gloeocystidiellumleucoxantha Russulacompacta 38 Lentinellusomphalodes E- Auriscalpiumvulgare 53~- Clavicoronapyxidata --- Bondarzewiaberkleyi .-- Heterobasidionannosum Laxitextumbicolor Fistulina hepatica 50 Clavariaacuta ' _I Clavulinopsisfusiformis 100 r Macrotyphulacf. uncea Typhulaphacorhiza .....5 Pterula aff. epiphyltoides 24 95 Lentinulalateritia euagaric 30 Gerronemachrysophyllum -0 -_Pleurotus tuberregium Hygrocybeconica 2i-- Hygrophorusebureus______Albatrellussyringae Panus rudis Meripilusgiganteus *------Fomitopsispinicola _ 72 i Sparassis spathulata r Clavariazollingeri Clavariazollingeri ;pistillaris Isunicolor

gomphoid- chiastic phalloid Gloeocantharelluspurpurascens 64 Sphaerobolusstellatus Geastrumsaccatum Pseudocolusfusiformis Trametessuaveolens Laetiporussulphureus Botryobasidiumsubcoronatum Uantnareuus_11 L 1t cwarius i-- Cantharelluslutescens - Craterellusfallax Cantharellustubaeformis Cantharellussp. Craterelluscornucopioides ,/\ Hydnumrepandut n Clavulina cinerea ,., O0r- 49 e, l"LavtUlavut ina cristata 6060 i. L r Clavulilutav?u na ornatipes Stichoclavariamucida stichic 87 99 Bjerkanderaadusta - E- Phanerochaetef chrysosporium Oxyporussp. Schizoporaparadoxa Coltriciaperenis Tremellasp. - Phellinus igniarius

FIG. 3. Phylogram of combined data for mit-ssu rDNA and nuc-ssu rDNA. One of 4458 equally parsimonious trees, see = Methods for analysis parameters. L (length) = 2022, CI (consistency index) = 0.380, RI (retention index) 0.606, RC (rescaled consistency index) = 0.231. See FIG. 1 for explanation of conventions. In this figure, dashed branches indicate alternate placements of the branch leading to Sparassis spathulata and Clavaria zollingeri (arrow), which are reflected in FIGS.1, 2. Line drawings (after 41) depict stichic vs chiastic meiotic division in immature basidia.

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions PINE ET AL: CANTHARELLOID AND CLAVARIOID PHYLOGENY 955

100 Dacrymyceschrysospermus '-- Auriculariaauricula Bondarzewiaberkleyi Heterobasidionannosum Echinodontiumtinctorium Lentinellusomphalodes | 53 -- Clavicoronapyxidata Russula compacta Peniophoranuda ~100 ~ I- Stereumhirsutum - - ---100 - J Gloeocystidiellumleucoxantha Aleurodiscusbotryosus Fistulina hepatica 50 Clavariaacuta "- _-- Clavulinopsisfusiformis Q100 - Macrotyphulajuncea Typhula phacorhiza Pterula dff. epiphylloides eul 95 Lentinulalateritia agaric Gerronemachrysophyllum Pleurotustuberregium Hygrocybeconica Hygrophoruseburneus Laxitextumbicolor Boletus satanas 72 Sparassis spathulata _, I 100 - Clavaria zollingeri Clavariazollingeri Meripilusgiganteus Panus rudis Trametessuaveolens Laetiporussulphureus 99 -- Bje randera adusta 1- Phanerochaetechrysosporium ------_------Fomitopsis pinicola Albatrellussyringae Botryobasidionsubcoronatum Schizoporaparadoxa ---.------.------Hyphodontia alutaria Oxyporussp. Clavariadelphuspistillaris Clavariadetphusunicolor Gloeocantharelluspurpurascens 100 j-- Gomphus bonarii "11 Gomphusfloccosus 100 64 f Sphaerobolusstellatus gomphoid- - eastrumsaccatum Pseudocolusfusiformis phalloid Clavariadelphusligulus Ramariaformosa Ramaria stricta Lentaria bvssiseda Cantharelluscibarius--- 100 Cantharelluslutescens 100 -- Cantharellussp. 100 Cantharellustubaeformis Craterellus cornucopioides Craterellusfallax sti ichic Hydnumrepandum 100 Clavulina cinerea 66 ? -- Clavulina cristata I Clavulina ornatipes ------Stichoclavaria mucida Coltriciaperenis Tremellasp. Phellinus igniarius

FIG. 4. Strict consensus tree for combined data. Consensus of 4458 equally parsimonious trees, L eq 2022, normalized CFI (consistency fork index) = 0.422. See FIG. 1 for explanation of conventions. Dashed branches indicate alternate place- ments of the branch leading to Sparassis spathulata and Clavaria zollingeri (arrow).

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions 956 MYCOLOGIA

Cantharelluscibarius 71.-Clavariadelphus ligulus 100 100- Cantharelluslutescens _LrClavariadelphus unicolor 100 t- Cantharellussp. -9- Clavariadelphus pistillaris 71LCantharellus tubaeformis - Ramaria stricta - Craterellusfallax Ramariaformosa steps * Craterelluscornucopioides 96 - Hydnumrepandum - i- Gomphus floccosus 99Gomphus bonarii Stichoclavaria mucida Gloeocantharellus 99r Clavulina cinerea purpurascens 91 rt I I Pseudocolus Clavulina cristata fusiformis Clavulina 30 steps q 66 ornatipes ~I stellatus Sphaerobolus FIG. 6. One of the two most parsimonioustrees for re- FIG. 5. most for re- aligned combined data of stichic taxa.L = 419, CI = 0.924, Single parsimonious phylogram = = combined data of clade. L = RI 0.921, RC 0.850. See FIG.1 for explanationof con- aligned gomphoid-phalloid ventions. is 332, CI = 0.782, RI = 0.557, RC = 0.406. See FIG. 1 for Topology congruentwith some of the most par- simonioustrees for all taxa. Rooted after explanation of conventions used in figure. Topology is con- analysesincluding FIG. 3. gruent with some of the most parsimonious trees for anal- yses including all taxa (e.g., FIG.3). Rooted after FIG.3 and Hibbett et al (37). of omitting large numbers of characters vs including characters that may add noise or be positively mis- clade in the most parsimonious tree, bootstrapping informative. However, results from analyses of the does not support monophyly of Gomphus and Ra- three exclusions sets suggest that these factors did maria. The genus Ramaria appears to be paraphylet- not affect conclusions of this study, since all phylo- ic. Clavariadelphuspistillaris and C. unicolor are sister genetic resolution receiving even moderate bootstrap taxa, as are C. ligulus and Lentaria byssiseda;Clava- support from any of the three exclusion sets tested is riadelphus is monophyletic if L. byssisedais included. compatible with results from all three. The Clavariadelphus lineage is nested within Gom- Possible causes for incongruence of the underlying phaceae, although bootstrap support is weak. Gloeo- phylogeny of two genes from the same taxa include: cantharellus appears to be the basal lineage within incomplete lineage sorting, hybridization, and other Gomphaceae, but its position is not supported by modes of horizontal transfer (54). The first two phe- bootstrapping. Pseudocolus, Geastrum, and Sphaero- nomena most likely occur among closely related spe- bolus are weakly supported as the monophyletic sister cies; this study focuses on relationships among gen- group of Gomphaceae. era and families that presumably diverged long ago. Among stichic taxa, Cantharellus lutescens and C. Horizontal transfer of the genomic regions used in tubaeformisform a clade which is the sister group of this study has never been reported. Thus, we expect- Craterellusfallax and C. cornucopioides(FIG. 6). There ed the mt-ssu-rDNA and nuc-ssu-rDNA sequences of is strong support (100%) for the monophyly of these the taxa in this study to represent the same under- taxa, to the exclusion of Cantharellus cibarius. Hyd- lying phylogeny. This expectation was supported by num repandum is the sister group of Cantharellaceae. comparison of bootstrap values for the two gene phy- Unfortunately, attempts to amplify DNA isolated logenies (FIGS. 1, 2)-all positive conflict between from Pseudocraterelluswere unsuccessful. Clavulina is the two trees receives less than 70% bootstrap sup- monophyletic and is the sister group of Stichoclava- port in at least one gene phylogeny. ria. The most distinctive topological conflict between the two gene trees concerns the placement of Cla- varia and Mitochon- DISCUSSION zollingeri Sparassis spathulata. drial data depict these taxa as monophyletic, and In many cases, inclusion of additional characters in place them within a clade that is otherwise stichic phylogenetic analysis increases the probability of cor- (FIG. 1). This differs from their placement based on rectly estimating the underlying tree topology (87). nuc-ssu-rDNA (FIG. 2) or morphological characters, But inclusion of ambiguously-aligned regions intro- but strong support for any of these placements is duces characters whose homology is questionable. lacking. Previous analyses of 1.2 kb of nuc-ssu-rDNA Furthermore, inclusion of phylogenetically informa- and the MS1/MS2 fragment of mt-ssu-rDNA support tive characters for which multiple taxa are scored as monophyly of Sparassis, , and missing sometimes can result in spurious resolution Laetiporus sulphureus, but only with moderate (50%) of artificial clades during parsimony analysis (53). bootstrap support (35). However, when additional Thus, there is a dilemma in phylogeny reconstruction taxa are sampled and complete nuc-ssu-rDNA se-

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions PINE ET AL: CANTHARELLOID AND CLAVARIOID PHYLOGENY 957 quences (1.8 kb) are included, bootstrap support for of Hydnum repandum receives unequivocol support this clade rises to 98% (37). Sparassis, Laetiporus,and in analyses with more evenly distributed branch Phaeolus all have ellipsoid-ovoid, smooth, inamyloid lengths (FIGS. 1, 3). spores, produce a brown rot, and can cause root and Overview.-Higher level relationships of Homobasid- butt rot of living trees, although host ranges differ. iomycetes are not resolved by these analyses (FIG.4). Taken together, the ecological and anatomical char- Nevertheless, in no analyses do cantharelloid and cla- acters and nuc-ssu-rDNA evidence suggest that the varioid fungi appear to form a basal, paraphyletic correct placement of Sparassis is with Laetiporus.The group from which the rest of the lengths of the branches leading to Clavaria zollingeri, have been derived. Thus, this sttdy provides no sup- Sparassis, and their putative sister taxa (see Results) port for Corner's Clavaria theoty of Homobasidi- in both the combined tree (FIG.3) and the mt-rDNA omycete evolution. Additionally, there is no evidence gene tree (FIG. 1) suggest that long branch attraction for a relationship between Cantharellaceae and the could be responsible for their placement. In certain agarics Gerronema or ,supporting Donk's cases of grossly unequal branch lengths, parsimony (19) and Heinemann's (33) conclusions that similar- analysis has been demonstrated to artificially connect ities of these genera to Cantharellaceae are due to extremely long branches that are unrelated in the convergence. Instead, it appears that many clavarioid true underlying phylogeny (27, 40). The branch lead- fungi, traditionally placed in Clavariaceae, are de- ing to Stichoclavaria is the longest in the mt-ssu-rDNA rived from a lineage (designated euagaric) that also tree (FIG. 1) if C. zollingeri and Sparassis are deleted gave rise to the gilled mushrooms Lentinula, Pleuro- (55 steps), suggesting that it is a likely candidate for tus, Hygrophorus, Hygrocybe,and Gerronema,and to this analytical artifact. Furthermore, monophyly of the polypore Fistulina. stichic taxa, C. zollingeri, and Sparassis receives only Coral- and club-shaped fungi have been derived in marginal (62%) bootstrap support, while monophyly four lineages, two of which have also given rise to of stichic taxa receives 92% bootstrap support in anal- cantharelloid fungi. The fruiting bodies of the near- yses of mt-ssu-rDNA that exclude the problematic C. est extant relatives of the different lineages represent zollingeri and Sparassis. Thus the mitochondrial data a wide range of forms: gilled mishrooms, toothed alone do not unambiguously support the placement fungi, , stinkhorns, and the cannon-ball fun- of C. zollingeri and Sparassis, and furthermore, un- gus. Similar rapid evolution of fruiting-body macro- derlying phylogenetic signal supports monophyly of morphology has been documented in diverse line- stichic taxa. Although branch lengths are more even- ages of Homobasidiomycetes (7, 37, 38, 47, 59). The ly distributed in combined analyses (FIG.3), Sparassis agarics lepideus and Lentinellus can pro- and C. zollingeri are still extremely divergent and are duce clavarioid fruiting bodies under appropriate en- grouped together. There is no support for their vironmental conditions (9, 57, 61 p 184), and Donk placement in the tree, resulting in three alternative (19 p 207-208) discusses several taxa whose fruiting placements (FIGS.3, 4) and lack of resolution in the bodies can be either corticioid or clavarioid. It ap- strict consensus of equally parsimonious trees (FIG. pears that superficial similarity of form is not a good 4). In the most conservative estimate, data presented predictor of evolutionary proximity in the cantha- in this study are insufficient to resolve relationships relloid and clavarioid fungi. Instead, our results sug- of Clavaria zollingeri and Sparassis. However, if evi- gest that certain anatomical featires are conserved dence from anatomical and ecological characters and within lineages that are otherwise morphologically nuc-ssu-rDNA are given precedence over the dubious diverse. For the coralloid Clavicorona, the mt-rDNA results, Clavaria with Cla- example, zollingeri belongs toothed and the Lentinellus form varia acuta and is the sister Auriscalpium, gilled Clavulinopsis, Sparassis a characterized of and neither are nested within monophyletic group by amyloid group Laetiporus, ornamentation. One of the of this the stichic clade. spore goals study was to evaluate putative synapomorphies for other A similar argument can be used to explain the po- lineages with cantharelloid and clavarioid members. lyphyly of stichic taxa found in the nuc-ssu-rDNA analyses (FIG. 2). The longest branches in the tree Gomphoid-phalloid clade.-Gomphus, Ramaria, and are found within the Cantharellaceae, and the only Gloeocantharellus are united by cyanophilic, warty other branch that is nearly as long is at the base, spore ornamentation and by green reactivity to iron leading to Dacrymyces. The rest of the stichic clade salts (TABLE I). Gloeocantharellushas a cantharelloid exhibits extremely short branches. We conclude that aspect, but has true gills and contains abundant Cantharellaceae is probably drawn to a basal position gloeoplerous hyphae. Our results strongly support in analyses based on nuc-ssu-rDNA because of its high the accepted placement of Gloeocantharellus with degree of divergence. Its position as the sister group Gomphus and Ramaria, indicating that Gloeocantha-

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions 958 MYCOLOGIA rellus represents an independent derivation of gills byssisedaappears to be nested within Clavariadelphus within the gomphoid-phalloid lineage, and is unre- rather than Ramaria (FIG.5). lated to any other agaric or boletoid fungi examined Several other taxa have been reported to stain thus far. green on contact with iron salts. Petersen (68, 72) Other fungi reported with spores and macrochem- described green or gray-green reactions of Cantha- ical reactivity similar to Gomphaceae include hyd- rellus cibarius and some species of Clavulinopsis noid Beenakia (44, 52, 60), and the resupinates Ka- (which he redefined as Ramariopsis in 1978), but lat- vinia, which is toothed, and Ramaricium, which has er (78) reported that pistillarin, the compound re- a smooth hymenophore (19, 24, 52). Although these sponsible for the green reaction in Gomphaceae and taxa are not represented in this study, sequences Clavariadelphus, was not present in Clavulinopsis. from the mitochondrial large subunit rDNA support Our results suggest that neither Cantharellusnor Cla- placement of Kavinia with Gomphus and Ramaria vulinopsis are related to Gomphaceae, indicating that (8), suggesting that spore morphology and iron salt green iron salt reactions in the absence of pistillarin reactivity may be a synapomorphy of this group. An- are not phylogenetically informative. Welden (90) other relative of Gomphaceae seems to be , suggested that Stereum radicans (= StereopsisReid) a false-truffle with striate, brown-pigmented spores was related to Gomphaceae and Clavariadelphus, that are also reported to be cyanophilous (46). Se- since it also stains green on contact with iron salts, quence data from mitochondrial large subunit rDNA but its pistillarin content has not been examined and (8) and nuclear large subunit rDNA (J. Spatafora it was not represented in this study. pers comm) support placement of Gautieria with The remaining taxa in the gomphoid-phalloid Gomphaceae. Petersen (71 p 15) reported that "the clade are Pseudocolus, Geastrum, and Sphaerobolus. staining reaction and general construction of the Bootstrap support for the placement of these gaster- spore wall" of Gymnopilusand the boletoid taxa, Por- omycetes with Gomphaceae is unequivocal (100%), phyrellus subflavidus, Strobilomycesconfusus, and S. and inclusion of the rest of the nuclear 18S rDNA floccopus are very similar to that of Gomphus, but re- and additional taxa does not alter this result (37). lationships among these taxa and Gomphaceae have Furthermore, sequences from nuclear large subunit not been examined further. rDNA (28S) support a relationship between Gom- Members of the club-shaped genus Clavariadelphus phales and (J. Spatafora pers comm). Re- react green on contact with iron salts (TABLEI), re- lationships among stinkhorns, earth-stars, the can- flecting the presence of pistillarin (56). These anal- non-ball fungus, and cantharelloid and clavarioid yses provide 100% bootstrap support for the place- fungi have never been proposed in the taxonomic ment of Clavariadelphus within Gomphaceae, reject- literature, and no morphological synapomorphy has ing a relationship with Cantharellaceae and with Cla- yet been identified for this diverse clade. Although variaceae. Although Clavariadelphus spores are Pellegrini and Patrignani's (62) examination of sep- smooth, hyaline, and unreactive, like those of Can- tal pore apparatuses let them to suggest that "the tharellaceae and Clavariaceae, this state appears to be genus Clavariadelphuscould be placed closer to Phal- plesiomorphic conservation of ancestral features. lales owing to the perforate parenthesome with small is a Lentaria in the restricted sense homogenous irregular holes," they observed intact dolipore septa of clavarioid char- group branched, lignicolous fungi in all Ramaria species examined. Fungi in the gom- smooth and thick-walled acterized by white, spores phoid-phalloid clade are remarkably ecologically and that the a leath- generative hyphae give fruiting body morphologically diverse, and have traditionally been texture Corner included in the ery (67, 72). (10, 14) examined by different groups of mycologists. Com- stichic that Petersen genus phycophilous, species parative studies of the anatomy and biochemistry of into Multiclavula (67) segregated (= Stichoclavaria). these taxa might elucidate morphological features Corner left Lentaria in Clavari- Although (10 p 24) that unite the lineage, and should be pursued. For he noted a resemblance between L. aceae, byssiseda example, iron salt reactions and pistillarin content of and the Stricta of shared re- group Ramaria; green Pseudocolus, Sphaerobolus,and Geastrumshould be in- actions with iron salts and thick-walled skeletal hy- vestigated. phae led Petersen (65, 72 Fig. 10) to conclude that Lentaria s. s. was derived within Ramaria. Corner Stichic clade.-Although monophyly of stichic taxa is (14) moved Lentaria from Clavariaceae into a new not supported by all analyses, placement of Sparassis family, Ramariaceae. Our results support Petersen's and Clavaria zollingeri in the midst of an otherwise separation of Multiclavula (= Stichoclavaria) and stichic clade is difficult to accept. Because such a re- Lentaria s. s., as well as the placement of the latter lationship is contradicted by all evidence except mt- genus in Gomphaceae (FIG. 3), although Lentaria ssu-rDNA sequences, which may be susceptible to

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions PINE ET AL: CANTHARELLOID AND CLAVARIOID PHYLOGENY 959 long branch attraction of these taxa and do not pro- it differs only in having a weaker carotenogenesis, vide strong bootstrap support, we reject the mt-ssu- with correspondingly relatively strong development rDNA results for Sparassis and C. zollingeri in favor of dark pigments of another sort." In contrast, Fias- of the placements supported by nuc-ssu-rDNA. Simi- son et al (30) found that Cr. cornucopioideswas totally larly, the removal of Cantharellaceae from the stichic devoid of carotenoids, while Cr.fallax, which is oth- clade to a basal position in the tree, seen only in the erwise very similar to Cr. cornucopioides,possessed the nuc-rDNA analyses (FIG.2), can be explained by long same carotenoids as C. cibarius. It is intriguing that branch attraction. Thus we conclude that stichic taxa carotenoid pigment structure seems to correlate with form a monophyletic group, Sparassis and Laetiporus, relationships supported by other characters, but until both brown rot fungi, are sister taxa, and the genus more taxa are examined and conflicting reports are Clavaria is most likely monophyletic and nested with- resolved it is impossible to determine the pattern of in the euagaric clade. pigment evolution within Cantharellaceae. Our data Our results provide strong (100% bootstrap) sup- do support multiple derivation of bicyclic caroten- port for the monophyly of Cantharellaceae, but Can- oids in diverse lineages, since neither Gerronemanor tharellus as previously defined appears to be paraphy- Clavulinopsis are closely related to any members of letic (FIG. 6). Cantharellus cibarius is the sister taxon Cantharellaceae. of a clade consisting of C. tubaeformis,C. lutescens (= Hydnum repandum is the sister group of Cantha- xanthopus, see 20), Craterellusfallax, and Cr. cornu- rellaceae, supporting Donk's (17, 19) and Petersen's copioides. These results confirm earlier findings by (72) conclusions based on morphological similarity. Feibelman et al (26) and Dahlman et al (pers Note that we are using Donk's (19) restricted defi- comm). Feibelman et al (26) recently proposed a nition of Hydnum, typified by H. repandum;the name new circumscription of genera within Cantharella- Dentinum, which has been used for this group, is in- ceae based on results from phylogenetic analyses of valid (76). Although nuc-ssu-rDNA data taken alone nuclear large subunit (28S) rDNA sequences. Feibel- remove Cantharellaceae from the stichic clade (FIG. man et al included only three of the species in our 2), the extreme divergence of the nuclear rDNA of study, but their conclusion that a clade containing C. Cantharellaceae (18S: note the long branches in FIG. cibarius can be separated from a clade including Cr. 2; ITS: ref. 25; 28S: J. Spatafora pers comm) makes fallax and C. tubaeformisis concordant with our re- it very likely that long branch attraction is responsible sults (FIG.6). They revised Cantharellus to contain C. for the placement of Cantharellaceae near the base cibarius and its relatives, and suggested that the genus of the tree and for the absence of support from boot- Craterellusbe expanded to include C. tubaeformisand strapping when the nuc-ssu-rDNA is taken alone. The Pseudocraterellus sinuosus, in addition to traditional branch length disparity of Cantharellaceae is much members of Craterellus(e.g., Cr fallax, Cr. odoratus). less severe in combined analyses, which provide un- If our results are fitted to their generic circumscrip- equivocal (100% bootstrap) support for the mono- tion, C. lutescens must also be transferred to Crate- phyly of Cantharellaceae and Hydnum (FIG. 3). Al- rellus. Feibelman et al also evaluated some of the though Hydnum has a toothed hymenophore, it is morphological features discussed in the Introduc- similar to Cantharellus in color, aspect, anatomy, and tion, and concluded that "shape and texture seem to flavor (TABLEI), and also has stichic nuclear division be more important [characters] than clamps, second- (80). ary septa, development, or hymenial configuration" Stichic nuclear division (see FIG. 3) was first de- in evaluating relationships of Cantharellaceae. scribed byJuel (41), soon after which Maire (55) pro- Analyses of carotenoid pigments of Cantharella- posed a classification scheme for the fleshy basidio- ceae provide some support for the circumscription mycetes based on the distinction between stichic and suggested by Feibelman et al (26). Cantharellus lutes- chiastic basidia. Ulbrich (88) erected new genera and cens, C. tubaeformis, and other members of Cantha- families for stichic taxa, but his classification was rellus subg. Phaeocantharellus sensu Corner (12) ac- largely ignored by subsequent literature. Many au- cumulate carotenoids with aliphatic structure exclu- thors since have criticized the use of this character sively, while C. cibarius and other members of Cor- for (10 p 27, 12 p 11, 43, 66, 83). Although ner's subgenus Cantharellus (roughly corresponding Donk's early work (17) gave strong weight to stichic to genus Cantharellus sensu 26) accumulate predom- nuclear division, placing Clavulina and Hydnum s. s. inantly bicyclic carotenoids (2, 28). However, pub- within Cantharellaceae, he later (18, 19) revised his lished reports of pigment analyses provide conflicting opinion, removing Clavulina to its own family on the results in some cases. For example, Arpin and Fiasson grounds that it was so evolutionarily divergent that its (2 p 84) state that "C[r]. cornucopioideslinks closely nearest relatives could not be determined. Our re- to the group C. lutescens-C. tubaeformis,from which sults support Maire's (55), Ulbrich's (88), and Donk's

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions 960 MYCOLOGIA original (17) concepts of close relationships among phyletic in most analyses, although without strong all stichic taxa. bootstrap support. A clade including these clavarioid Petersen (67) segregated small, lichenized, un- fungi and Pterula, the mushrooms Hygrocybe,Hygro- branched clavarioid fungi into the genus Multicla- phorus, Pleurotus, Gerronema,Lentinula, and the poly- vula, but suggested that Multiclavula belonged in a pore Fistulina appears in all of the most parsimoni- generic complex with Clavaria (72). Hubbard and ous trees from the combined data (FIG.4). Although Petersen (39) concluded that Juel (42) was likely ex- this clade does not receive strong bootstrap support amining a Multiclavula when he described the nucle- (30%), analyses including more non-cantharelloid or ar state of Clavaria falcata. In 1928, Ulbrich erected clavarioid taxa and the rest of the nuc-ssu-rDNA pro- the family Stichoclavariaceae, including two genera- vide 97% bootstrap support for the placement of Ty- Stichoclavaria, typified by C. falcata, and Stichorama- phula and Fistulina with Pleurotus, Lentinula, and ra, including S. rugosa, S. cristata, S. cineria, and S. other members of the euagaric clade (37). Future stichic clavarioid grisea-for fungi. Although Ul- mycological studies cannot assume that mushrooms brich's Stichoramariais a for the older Cla- synonym and coral and club fungi represent distinct lineages. we concur with Hubbard and Petersen's vulina, sug- Furthermore, it is now clear that coral and club fungi that "Stichoclavaria should be reconsidered gestion have been derived multiple times from diverse line- as the for the Multiclavula complex." ages, and do not represent an ancestral group that Our results Petersen's of support segregation gave rise to the more complex fruiting forms found Stichoclavariafrom other clavarioid but fungi, suggest in the Basidiomycetes. that similarities between Stichoclavaria and Clavaria are due to convergence; the nearest relatives of Stich- oclavaria are taxa with the same mode of meiotic nu- ACKNOWLEDGMENTS clear division. We thank Ammirati, Eric Danell, Dennis The stichic not Joe Desjardin, only reportedly genera represented Robert Ginns, David Ronald Pe- in this are the Clavulicium and Fogel,James McLaughlin, study resupinate fungi tersen, Donald Pfister,Barbara Thiers, and Qiuxin Wu for Sistotrema. Clavulicium is similar to anatomically very providingfungal material,Emily Wood and Donald Pfister Clavulina (5), while Sistotremapossesses unique ur- for curatorialsupport, Steve Rehner for technical advice, niform basidia that make its relationship to other ba- Joey Spataforaand MattiasDahlman for sharing unpub- sidiomycetes difficult to ascertain; no known data lished results,David Swofford for providingtest versionsof contradict a relationship with the stichic clade re- PAUP*,Tom Bruns,John Taylor,and two anonymousre- vealed by our analyses. Because such a wide range of viewers for helpful comments, and Toby Kellogg, David chiastic genera were sampled, it is likely that sticho- Baum,and membersof the Donoghue lab for their immea- surable advice and was Na- basidia are indeed uniquely derived and have never support. Funding provided by tional Science Foundation Grant DEB-9629427to been reversed. Still, nuclear behavior during meiosis MJ.D. has to be examined in of basidio- and D.S.H. and by Ford Foundationand Sigma Xi grants yet many groups to E.M.P. mycetes. Attempts to identify correlated characters, such as narrow, elongate basidia, have been strongly criticized (19 p 220). For example, Hygrocybeis ana- tomically very similar to stichic fungi, notably in ba- LITERATURE CITED sidial shape (34), but is reported to be chiastic (55). 1. VizziniA. 1994. Un Craterellustutto If stichobasidia are as phylogenetically informative as Apicella P, giallo: Craterelluskonradii R. Maire et Bourdot 1928. Rivista these results suggest, examination of more taxa may Micol 38:173-177. identify other relatives of Cantharellaceae, Hydnum, 2. N, Fiasson 1971. The of Basidio- Stichoclavaria, and Clavulina. Arpin J-L. pigments mycetes:their chemotaxonomicinterest. In: Petersen ed. Evolutionin the an in- Euagaric clade.-Although the evolutionary relation- RH, higher Basidiomycetes: ternational Knoxville: of Ten- ships of the remaining clavarioid genera are not de- symposium. University nessee Press.p 63-98. finitively resolved by these data, they appear to be 3. BerthierJ. 1974. Le genre Typhula(Clavariacees) et les nested within the lineage containing the major radi- genres affines classification-especes nouvelles. Bull ation of mushrooms (FIGS.3, 4). The mono- gilled Soc Linn Lyon 43:182-188. of and phyly Macrotyphulajuncea Typhulaphacorrhiza 4. BigelowHE. 1978. The cantharelloidfungi of New Eng- is well that supported (100% bootstrap), suggesting land and adjacentareas. Mycologia 70:707-756. earlier placements of Macrotyphulawith Clavariadel- 5. Boidin J. 1957. Heterobasidiomycetessaprophytes et phus (14, 43) were erroneous. Clavaria acuta and Homobasidiomycetesresupines. II. Catalogueraisonne Clavulinopsis (= Ramariopsis) fusiformis are mono- des especes pyreneennes de la region de Luchon

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions PINE ET AL: CANTHARELLOID AND CLAVARIOID PHYLOGENY 961

(Haute-Garonne). Bull Soc Hist Nat Toulouse 92:277- inferred from sequence analysis of the nuclear large 292. subunit rDNA. Mycol Res 101:1423-1430. 6. . 1958. Essai biotaxonomique sur les hydnes re- 27. Felsenstein J. 1978. Cases in which parsimony or com- supines et les corticies: etude speciale du comporte- patibility methods will be positively misleading. Syst ment nucleaire et des myceliums. Rev Mycol Memoire Zool 27:401-410. 6:1-387. 28. Fiasson J-L, Arpin N. 1967. Recherches chimiotaxino- 7. Bruns TD, Fogel R, White TJ, Palmer JD. 1989. Accel- miques sur les champignons V.-sur les carotenoides erated evolution of a false-truffle from a an- mineurs de Cantharellus tubaeformisFr. Bull Soc Chim cestor. Nature 339:140-142. Biol 49:537-542. 8. , Szaro TM, Gardes M, Cullings KW, Pan JJ, Tay- 29. , Bouchez M-P. 1968. Recherches chimiotaxino- lor DL, Horton TR, Kretzer A, Garbelotto M, Li Y. miques sur les Champignons. Les carotenes de Om- Rend Acad Sci Ser 1998. A sequence database for the identification of ec- phalia chrysophyllaFr. Compt Paris, D tomycorrhizal basidiomycetes by phylogenetic analysis. 266:1379-1381. Mol Ecol 7:257-272. 30. , Petersen RH, Bouchez M-P, Arpin N. 1970. 9. Buller AHR. 1905. The reactions of the fruit-bodies of Contribution biochimique a la connaissance taxino- de certains cantharelloides et cla- lepidus, Fr., to external stimuli. Ann Bot 19: mique champignons 427-438. varioides. Revue Mycol 34:357-364. 31. Harrison KA. 1971. The lines in the 10. Corner EJH. 1950. A monograph of Clavaria and allied evolutionary fungi with the In: Petersen genera. Ann Bot Mem 1:1-740. spines supporting hymenium. ed. Evolution in the an in- 11. . 1957. Craterellus Pers., Cantharellus Fr., and RH, higher Basidiomycetes: ternational Knoxville: of Ten- Pseudocraterellusgen nov. Sydowia, Beih Ser II 1:266- symposium. University 276. nessee Press. p 375-392. 32. Heim R. 1954. A de trois chanterelles americai- 12. . 1966. A monograph of cantharelloid fungi. Ann propos nes. Revue 19:47-56. Bot Mem 2:1-255. Mycol 33. Heinemann P. 1961. Les criteres chez les 13. . 1970a. Notes on cantharelloid fungi. Nova Hed- systematiques Cantharellineae. 15:200-203. 29:783-818. Sydowia wigia 34. Hesler Smith AH. 1963. North American of 14. . 1970b. to "A of Cla- LR, species Supplement monograph Knoxville: of Press. varia and allied Beih Nova 33:1- Hygrophorus. University genera." Hedwigia 416 299. p. 35. Hibbett DS. 1996. evidence for horizontal 15. . 1972. Boletus in Botanic Phylogenetic Malaysia. Singapore: transmission of I introns in the nuclear ribosom- Gardens. 263 group p. al DNA of Mol Biol Evol 13: 16. , Thind KS. 1961. Dimitic of Ramaria mushroom-forming fungi. species 903-917. (Clavariaceae). Trans Br Soc 44:233-238. Mycol 36. , MJ. 1995. toward a 17. Donk MA. 1933. Revision der Niederlandischen Hom- Donoghue Progress phylo- genetic classification of the through par- obasidiomycetae-Aphyllophoraceae II. Mededeel Bot simony analysis of mitochondrial ribosomal DNA se- Mus Herb R Univ Utrecht 9:1-278. quences. Can J Bot 73:S853-S861. 18. . 1961. Four new families of Hymenomycetes. 37. , Pine EM, Langer E, Langer G, Donoghue MJ. Persoonia 1:405-407. 1997. Evolution of gilled mushrooms and puffballs in- 19. . 1964. A of the families of conspectus Aphyllo- ferred from ribosomal DNA sequences. Proc Natl Acad phorales. Persoonia 3:199-324. Sci USA 94:12002-12006. 20. . 1969. Notes on Cantharellus sect. Leptocantha- 38. Hopple JS Jr, Vilgalys R. 1994. Phylogenetic relation- rellus. Persoonia 5:265-284. ships among coprinoid taxa and allies based on data 21. . 1971. in the of the classification Progress study from restriction site mapping of nuclear rDNA. Mycol- of the In: Petersen ed. Evo- higher basidiomycetes. RH, ogia 86:96-107. lution in the higher Basidiomycetes: an international 39. Hubbard M, Petersen RH. 1979. Studies in basidial nu- Knoxville: of Tennessee Press. symposium. University p clear behavior of selected species of clavarioid and can- 3-25. tharelloid fungi. Sydowia, Bieheft 8:209-223. MS. 1947. a new the 22. Doty Clavicorona, genus among 40. HuelsenbeckJP. 1997. Is the Felsenstein zone a fly trap? clavarioid fungi. Lloydia 10:38-44. Syst Bio 46:69-74. 23. . 1948. A preliminary key to the genera of cla- 41. Juel HO. 1898. Die Kernthielungen in den Basidien varioid fungi. Bull Chicago Acad Sci 8:173-178. und die Phylogenie der Basidiomyceten. Jahrb Wiss 24. ErikssonJ. 1954. Ramaricium n. gen., a corticioid mem- Bot 32:361-388. ber of the Ramaria group. Svensk Bot Tidskr 48:188- 42. . 1916. Cytologische Pilzstudien I. Die Basidien 198. der Gattungen Cantharellus, Craterellusund Clavaria. 25. Feibelman T, Bayman P, Cibula WG. 1994. Length var- Nove Acta Reg Soc Sci Upsal Ser IV 4:1-40. iation in the internal transcribed spacer of ribosomal 43. Jiilich W. 1981. Higher taxa of Basidiomycetes. Biblioth DNA in chanterelles. Mycol Res 98:614-618. Mycol 85:1-485. 26. , Doudrick RL, Cibula WG, Bennett JW. 1997. 44. , Star W. 1983. Ultrastructure of , I Phylogenetic relationships within the Cantharellaceae Beenakia. Persoonia 12:67-74.

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions 962 MYCOLOGIA

45. Keller J. 1978. Ultrastructure des parois sporiques des species and notes on the origin of Clavulina. Mycolo- Aphyllophorales. V. Les genres Cantharellus, Craterel- gia 59:39-46. lus, et Pseudocraterellus.Schweiz Z Pilzk 56:150-154. 67. . 1967c. Notes on clavarioid fungi. VII. Redefi- 46. Kotlaba F, Pouzar Z. 1964. Preliminary results on the nition of the Clavaria vernalis-C. mucida complex. Am staining of spores and other structures of Homobasi- Midi Naturalist 77:205-221. diomycetes in Cotton Blue and its importance for tax- 68. . 1968a. The genus Clavulinopsis in North Amer- onomy. Feddes Repertorium 69:131-142. ica. Mycol Mem 2:1-39. 47. Kretzer A, Bruns TD. 1997. Molecular revisitation of the 69. . 1968b. Notes on cantharelloid fungi. I. Gom- genus Gastrosuillus. Mycologia 89:586-589. phus S.F. Gray, and some clues to the origin of rama- 48. Kuhner R. 1928. Omphalia chrysophyllaFries Bull Soc rioid fungi. J Elisha Mitchell Sci Soc 84:373-381. Myco France 44 (Suppl):Plate 28. 70. . 1969. Notes on cantharelloid fungi. II. Some 49. . 1947. Le comportement nucleaire dans les ba- new taxa, and notes on Pseudocraterellus.Persoonia 5: sides urniformes et a plus de quatre spores de Sistro- 211-223. trema confluens Pers. Compt Rend Acad Sci 224:1068- 71. . 1971a. The genera Gomphusand Gloeocanthar- 1069. ellus in North America. Nova Hedwigia 22:1-144. 50. , Romagnesi H. 1953. Flore analytique des cham- 72. . 1971b. Interfamilial relationships in the clava- pignons sup6rieurs. Paris: Masson et Cie. 556 p. rioid and cantharelloid fungi. In: Petersen RH, ed. Evo- 51. Leathers CR, Smith AH. 1967. Two new species of cla- lution in the higher Basidiomycetes: an international varioid fungi. Mycologia 59:456-462. symposium. Knoxville: University of Tennessee Press. p 52. Maas Geesteranus RA. 1963. Hyphal structures in hyd- 345-374. nums. III. Proc Kno Nederl Akad Ser C 66:437-446. 73. . 1972. Notes on clavarioid fungi. XII. Miscella- 53. Maddison WP. 1993. Missing data versus missing char- neous notes on Clavariadelphus, and a new segregate acters in phylogenetic analysis. Syst Biol 42:576-581. genus. Mycologia 64:137-152. 54. . 1995. Phylogenetic histories within and be- 74. . 1973. Aphyllophorales II: the clavarioid and tween species. Monogr Syst Bot, Missouri Bot Gard 53: cantharelloid Basidiomycetes. In: Ainsworth GC, Spar- 272-288. row FK, Sussman AS, eds. The Fungi: an advanced trea- 55. Maire R. 1902. Recherches cytologiques et taxono- tise. :Academic Press. p 351-368. miques sur les Basidiomycetes. Bull. Soc. Mycol France 75. . 1975. Notes on cantharelloid fungi. VI. New 18:1-212. species of Craterellusand infrageneric rearrangement. 56. Methven AS. 1990. The genus Clavariadelphusin North Ceska Mykol 29:199-204. America. Biblioth Mycol 138:1-192. 76. . 1977. The typification of Hydnum Linn. per 57. Miller OKJr. 1971. The relationship of cultural char- Fries: time for stability. Taxon 26:144-146. acters to the taxonomy of the agarics. In: Petersen RH, 77. . 1978. Notes on clavarioid fungi. XV. Reorga- ed. Evolution in the higher Basidiomycetes: an inter- nization of Clavaria, Clavulinopsis and Ramariopsis. national symposium. Knoxville: University of Tennessee Mycologia 70:660-671. Press. p 197-215. 78. . 1985. Notes on clavarioid fungi. XIX. Colored 58. , Watling R. 1987. Whence cometh the Agarics? illustrations of selected taxa, with comments on Can- A reappraisal. In: Petersen RH, ed. Evolution in the tharellus. Nova Hedwigia Z Kryptogamenkunde 42: higher Basidiomycetes: an international symposium. 151-160. Knoxville: University of Tennessee Press. p 435-448. 79. Reijnders AFM, StalpersJA. 1992. The development of 59. Mueller GM, Pine EM. 1994. DNA data provide evi- the hymenophoral trama in the aphyllophorales and dence on the evolutionary relationships between mush- the agaricales. Stud Mycol 34:1-109. rooms and false truffles. McIlvanea 11:61-74. 80. Restivo JH, Petersen RH. 1976. Studies on nuclear di- 60. Nunez M, Ryvarden L. 1994. A note on the genus Bee- vision and behavior within basidia. I. Hydnum umbili- nakia. Sydowia 46:321-328. catum. Mycologia 68:666-672. 61. Pegler DN. 1983. The genus Lentinus: a world mono- 81. Singer R. 1945. New genera of fungi-II. Lloydia 8:139- graph. Kew Bull Addit Ser 10:1-281 144. 62. Pellegrini S, Patrignani G. 1994. Septal pore apparatus 82. . 1947. Coscinoids and coscinocystidia in Linde- in some clavarioid fungi: taxonomic and phylogenetic romyceslateritius. Farlowia 3:155-157. implications. Caryologia 47:131-139. 83. . 1986. The Agaricales in modern taxonomy. Koe- 63. Penancier N. 1961. Recherches sur l'orientation des fu- nigstein, Germany: Koeltz Scientific Books. 981 p. seaux mitotiques dans la baside des Aphyllophorales. 84. Smith AH. 1971. The origin and evolution of the Agar- Trav Lab (La Jaysinia) 2:57-71. icales. In: Petersen RH, ed. Evolution in the higher 64. Persson O, Mossberg B. 1995. Fynd av svart trumpets- Basidiomycetes: an international symposium. Knox- vamp med gul farg. Svenska Bot Tidskr 89:58-59. ville: University of Tennessee Press. p 481-504. 65. Petersen RH. 1967a. Evidence on the interrelationships 85. Swann EC, Taylor JW. 1993. Higher taxa of Basidiomy- of the families of clavarioid fungi. Trans Br Mycol Soc cetes: an 18S rRNA gene perspective. Mycologia 85: 50:641-648. 923-936. 66. . 1967b. Notes on clavarioid fungi. VI. Two new 86. Swofford DL. 1991. PAUP: Phylogenetic Analysis Using

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions PINE ET AL: CANTHARELLOID AND CLAVARIOID PHYLOGENY 963

Parsimony, v. 3.1. Champaign, Illinois: Illinois Natural Booth C, ed. Methods in microbiology. London: Aca- History Survey. demic Press. p 567-598. 87. , Olsen GJ, Waddell PJ, Hillis DM. 1996. Phylo- 90. Welden AL. 1966. Stereum radicans, Clavariadelphus genetic inference. In: Hillis D, Moritz C, Mable BK, and the Gomphaceae. Brittonia 18:127-131. eds. Molecular systematics. Sunderland, Massachusetts: 91. White TJ, Bruns TD, Lee S, Taylor JW. 1990. Amplifi- Sinaur Associates, Inc. p 407-514. cation and direct sequencing of fungal ribosomal RNA 88. Ulbrich E. 1928. Die hoheren Pilze. Kryptogamenfl An- genes for . In: Innis MA, Gelfand H, Snin- fanger 1:1-497. skyJJ, White T, eds. PCR protocols. San Diego, Califor- 89. Watling R. 1971. Chemical tests in agaricology. In: nia: Academic Press. p 315-322.

This content downloaded from 130.132.173.57 on Tue, 10 Jun 2014 12:51:21 PM All use subject to JSTOR Terms and Conditions