Lepiota in California: species with a hymeniform pileus covering

Else C. Vellinga1

111 Koshland Hall 3102, Department of Plant andMicrobial Biology, University of California atBerkeley, Berkeley, California 94720-3102

Abstract: Eight Lepiota species with a hymeniformpileus covering that are known in California arepresented and discussed. Lepiota phaeoderma isdescribed as new; this species is characterized by adark pileus surface, which splits open into smallpatches around the umbo, and the absence of anannulus. Lepiota neophana, a species with an annulusand a closed brown pileus surface, and L. lilacea, witha conspicuous dark annulus, are both recorded forthe first time west of the Rocky Mountains. Theidentity of L. neophana is reviewed and clarified basedon morphological and molecular data. The typecollection of L. rufipes was studied, and L. rufipeswas placed in synonymy with Cystolepiota seminuda. Akey to the species is given.

Key words: Agaricaceae, biodiversity, Callitropsismacrocarpa, nrITS and EF1-a sequences, taxonomy,type studies

INTRODUCTION

Lepiota (Pers.) Gray is represented in the highlydiverse state of California by at least 25 species, 30%

of which are still undescribed. This genus is lessspecies-rich than Leucoagaricus (Locq.) ex Singer(Vellinga 2004a), which also contains many unde-scribed species in California. Several Lepiota specieshave been described from the state in the past (e.g.Murrill 1912; Sundberg 1971, 1989; Vellinga 2007),but an overview of the genus is still lacking.

The group of species with a hymeniform pileuscovering was the focus of this article. Basidiocarps ofthese species are small, with a pileus covering thateither covers the pileus homogeneously or splits openinto small patches or flat scales, depending on thepresence or absence of an intercellular matrix thatbinds the individual cells. An annulus can be presentor absent; the spores are ellipsoid or bullet-shaped,and small (less than 8 mm long), and the spore wallsdo not react strongly with Melzer’s reagent andCongo red. One species has only one nucleus per

spore (Kuhner 1945, Vellinga and Huijser 1999),whereas the other species have two, as is usual inLepiota. Cheilocystidia are present or absent. Themorphological differences among the species areoften subtle (Vellinga and Huijser 1999), but nrITSsequence data support the morphologically recog-nized species. However species with a hymeniformpileipellis do not form a monophyletic group(Vellinga 2003).

This group has been accommodated in 1–3 sectionsbased on morphological characters, according tovarious authors. Singer (1986) put all species insection Cristatae (Kuhner ex Wasser) Bon, regardlessof spore shape, but Bon (1993) placed them in threesections (in two subgenera) by distinguishing specieswith bullet-shaped spores (in sect. Cristatae) fromspecies with ellipsoid spores in subgenus ParalepiotulaBon. He then divided this subgenus in two: sect.Integrellae (Kuhner ex Bon) Bon for species with apileus covering that does not split open and only onenucleus per spore and sect. Lilaceae Bon for specieswhose pileus covering breaks open into scales andhave binucleate spores.

Phylogenetic analyses based on molecular datafailed to resolve many of these differences (Vellinga2003). However they clearly showed that species withbullet-shaped spores and a hymeniform pileus cover-ing, such as L. cristata, belong with ellipsoid-sporedspecies and a similar pileus covering and not withspecies with spurred spores and a trichodermal pileuscovering. The bullet-shaped spores arose once in thisclade of ellipsoid-spored species; the more pro-nouncedly spurred spores in the group of specieswith trichodermal or cutis-like pileus covering (L.boudieri Bres., L. castanea Quel.) evolved indepen-dently.

The molecular data (Vellinga 2003) also did notsupport a separate section for L. rufipes, which differsfrom the other species in the uninucleate spores. Theisolated and unresolved positions of Lepiota lilaceaBres., L. ochraceofulva P.D. Orton and L. pyrochroaMalencon, outside the core group of species with ahymeniform pileus covering, came as a surprise(Vellinga 2003) because morphologically they arenot distinct in essential characters from the rest of thehymeniform species.

Despite several articles on species in this groupfrom California (Sundberg 1971, 1989; Vellinga2001a, b) naming problems still persist. The identityof L. neophana Morgan, described from Preston,Ohio, (Morgan 1906) is at the center of the problem

Submitted 16 Jul 2009; accepted for publication 24 Sep 2009.1 Corresponding author. E-mail: ecvellinga@comcast.net

Mycologia, 102(3), 2010, pp. 664–674. DOI: 10.3852/09-180# 2010 by The Mycological Society of America, Lawrence, KS 66044-8897

664

because at least three different interpretations of thisspecies have been published (Smith 1954, Sundberg1989, Bizio et al. 1993). This species again has beenstudied and is here redescribed from modernCalifornian collections, according to the originaldescription by Morgan (1906). As a result the speciesdescribed as L. neophana sensu Sundberg (1989) is inneed of a new name and is described as L.phaeoderma.

Seven of the eight species presented here co-occurin one small area, containing several Montereycypress (Callitropsis macrocarpa (Hartw.) D.P. Little;syn. Cupressus macrocarpa Hartw.; Hesperocyparismacrocarpa (Hartw.) Bartel) plantations on an east-facing slope south of San Francisco, and one of theseseven is not known from any other location inCalifornia. These Monterey cypress groves are notablefor being rich in species of Lepiota and Leucoagaricus(Vellinga 2004a).

The species are treated in checklist format inalphabetical order, with a nomenclator and referenc-es to descriptions and illustrations in the literature.Species new to science and the newly recorded speciesare fully described. Additional morphological infor-mation is provided for several species, including thenumber of nuclei per spore, an important diagnosticcharacter. An identification key is provided to aidrecognition of the species.

MATERIALS AND METHODS

Standard methods for describing basidiocarps were appliedwith the terminology of Vellinga and Noordeloos (2001).Color annotations in macroscopic descriptions are fromMunsell2 soil color charts (1975). The notation [115,7,6]means that measurements were made on 115 spores inseven samples in six collections. These abbreviations areused: L for number of lamellae, l for number of lamellulaebetween two lamellae, avl for average length, avw foraverage width, Q for quotient of length and width andavQ for average quotient. Herbarium material was used tocount the number of nuclei per spore. Nuclei were stainedwith a 49, 6-diamidino-2-phenylindole, dihydrochloride(DAPI) solution (1 mg/mL) and observed with a fluores-cence microscope with Ex 355–375 BPass and Em LP 397filters. Herbarium abbreviations follow Holmgren andHolmgren (1998). All collections are in UC unlessotherwise stated. The Latin description of the new specieshas been deposited in MycoBank. DNA sequences wereproduced with standard molecular methods; DNA wasextracted with a QIAGEN DNeasyH Blood and Tissue Kit.For nrITS sequences the primer pair ITS1F and ITS4 wereused both for PCR and sequencing (Gardes and Bruns1993); part of the elongation factor 1-a gene (EF1-a) wasamplified with primers EF1-983F and EF1-1567R accordingto the protocol of Rehner and Buckley (2005). PCRproducts were cleaned with 0.5 mL ExoSAP IT (USB Corp.,

Cleveland, Ohio) and cycled at 37 C for 45 min, followed15 min at 80 C. Sequencing was performed with Big Dyechemistry and an ABI PRISM 3100 Genetic Analyzer(Applied Biosystems, Foster City, California). Sequenceswere edited and contigs assembled with Sequencher 4.2.2(Gene Codes Corp., Ann Arbor, Michigan). The nrITSsequences were aligned by partial order alignment (Lee etal. 2002), and the EF1-a sequences were aligned with ClustalX 2.09 (Larkin et al. 2007); PAUP* 4.0d81 (Swofford 2002)performed the phylogenetic analysis. Five species represen-tative of the major clades in Lepiota were chosen forcomparison. Chamaemyces fracidus (Fr.) Donk serves asoutgroup because it is basal in the Agaricaceae (Vellinga2004b). Dermoloma inconspicuum Dennis was included inthe analysis because it recently was shown to belong to theAgaricaceae (Kropp 2008); it shares the hymeniform pileuscovering with the species treated here. All 30 newlyproduced sequences have been deposited in GenBank.(Accession numbers are listed with the collections undercollections examined and in FIGS. 1, 6.) Alignments aredeposited in TreeBase under provisional number SN4563.Sequence data were used only to establish that themorphological differences were in concordance withsequence differences, not to establish a phylogenetichypothesis for genus Lepiota or the species with a hymeni-form pileus covering.

TAXONOMY

1. Lepiota castaneidisca Murrill, Mycologia 4:232.1912.

Misapplied. Lepiota conspurcata p.p. sensu Murrill, NAm Fl 10(1):56. 1914; Lepiota cristata p.p. sensuSundberg, Mycotaxon 34:240. 1989.

Selected description.Vellinga, Mycotaxon 80:302–303.2001.Lepiota castaneidisca was described from California

by Murrill (1912), subsequently considered synony-mous with L. cristata (under L. conspurcata [Willd.]Morgan) by the same author (Murrill 1914), treatedas a species with ellipsoid spores by Sundberg (1967)and later as a synonym of L. cristata (Sundberg 1989).Vellinga (2001) realized that there was indeed a taxonthat differs from L. cristata in California with arounded red-brown pileus instead of a distinctlyumbonate pileus with orange-like tinges and usedthe name L. castaneidisca for it. The nrITS sequencesneatly support the recognition of two species (FIG. 1).L. cristata and L. castaneidisca are indistinguishablemicroscopically. The spores are slightly metachromat-ic in Cresyl blue and have two nuclei. Lepiotacastaneidisca is known from California north intoWashington (Birkebak pers comm).

2. Lepiota cristata (Bolton : Fr.) P. Kumm., Fuhrer indie Pilzkunde:137. 1871.

; Agaricus cristatus Bolton, A history of fungusses

VELLINGA: LEPIOTA IN CALIFORNIA 665

FIG. 1. One of 10 000 most parsimonious trees based on a dataset of 70 nrITS sequences of Lepiota species with a hymeniformpileus covering, five species representative of the major clades in Lepiota (indicated by *), Dermoloma inconspicuum and Chamaemycesfracidus as outgroup (76 in total). Two hundred seventy-one characters were parsimony informative. Species occurring in California arehighlighted; the other hymeniform species are from Europe, with the exception of D. inconspicuum from Central and South America.Numbers above branches indicate branch length, numbers in italics below branches are an indication of the bootstrap support.Bootstrap support was calculated based on 50 000 replicates done in fast-stepwise-addition, with groups with . 70% support retained.

666 MYCOLOGIA

growing in Halifax 1:7. 1788, non Agaricus cristatusScop., 1774.

; Agaricus cristatus Bolton : Fr., Systema mycologi-cum 1:22. 1821.

; Lepiotula cristata (Bolton : Fr.) E. Horak, SynopsisGenerum Agaricalium:338. 1968.

Selected description. Vellinga, Flora agaricina neerlan-dica 5:138. 2001.Lepiota cristata is a widespread species complex

known from the temperate northern hemisphere(Vellinga 2001b; Liang, Xu and Yang 2009). nrITSsequences show considerable variation in this specieseven within collections from the same area (e.g. thethree collections from Ann Arbor, Michigan, fall intotwo clearly different groups based on nrITS sequencesand a small sample from California shows threedistinct nrITS types [Vellinga 2001b]), but a correla-tion with morphology has not been made, and at thispoint it is unclear whether these nrITS variantsrepresent different species. Several infraspecific taxahave been described, but because their status has tobe confirmed with molecular data they are not listedhere. Note that the description by Sundberg (1989)includes L. castaneidisca and hence is not listed underselected descriptions.

Lepiota kauffmanii was described as having a ‘‘Marsbrown’’ (dark brown) pileus that starts out campan-ulate, expanding to nearly plane umbonate (Zeller1933), and was considered synonymous with L.cristata (Smith 1966). The data on the shape of thepileus are not sufficient to determine this unambig-uously because they could refer to L. castaneidisca.The microscopical data fall within the variability ofboth L. castaneidisca and L. cristata, and a decisionon its synonymy cannot be made. The following notes

were made on the type collection (FIG. 2) (andcompare with Smith 1966). Basidiospores [25,2,1]5.4–6.9 3 3.2–4.2 mm, avl 3 avw 5 6.0–6.3 3 3.6–3.7 mm, Q 5 1.45–1.85, avQ 5 1.64–1.69, notdextrinoid, not metachromatic in Cresyl blue. Lamel-la edge sterile, with cheilocystidia forming a sterileband; cheilocystidia colorless, 20–42 3 9–13 mm,sphaeropedunculate to clavate with short to longpedicel, rarely narrowly clavate.

Lepiota cristata grows in relatively nutrient-richplaces ranging from natural habitats (especially riverbeds and other riparian habitats with regular flood-ing) to ruderal places with strong human influences.Human influence may account for the co-occurrenceof several nrITS types within a certain area.

Collection examined. USA. Oregon: Benton County,Alsea Mountain, 26 Oct 1930, S.M. Zeller 7806 (holotypeof Lepiota kauffmanii, NY).

3. Lepiota lilacea Bres., Fungi tridentini 2:3. 1892.FIG. 3

; Lepiotula lilacea (Bres.) Wasser, Nov Sist vyssh nizshRast 1975:191. (‘1975’) 1976.

5 Lepiota lilacea var. pallida Bon, Migl. & Cherubini,Doc mycol 19 (76):71. 1989.

5 Lepiota lilacea f. pallida (Bon, Migl. & Cherubini)Migl., Parliamo di Funghi:4. 2000.

5 Lepiota lilacea f. micropholoides Migl. & Coccia,Micol ital 21 (2):40. 1992.

Selected description.Vellinga, Flora agaricina neerlan-dica 5:140–141. 2001.

FIG. 2. Lepiota kauffmanii. A. Spores. B. Cheilocystidia.C. Pileus covering elements. All from the holotype (Zeller7806). Bar 5 10 mm.

FIG. 3. Lepiota lilacea. A. Spores. B. Basidia. C. Cheilo-cystidia. D. Pileus covering elements. All from J. Brown, 28Aug 2008. Bar 5 10 mm.

VELLINGA: LEPIOTA IN CALIFORNIA 667

Pileus 31–36 mm, 8–11 mm high, plano-convex toapplanate with glabrous dark violet disk, around diskbreaking open into radially arranged appressed scales,concolorous with disk; surface white and radially fibrilloseat margin and there devoid of scales. Lamellae, L 5 32–40, l5 1, free, crowded, broadly ventricose, up to 6 mm wide,white, with white, even edge. Stipe 23–29 3 6–7 mm,cylindrical to slightly widening downward, covered in awhite fibrillose layer, reddish purple in lower part underthis layer, in one specimen with some velar remnants similarto scales on pileus, in lower half hollow. Annulus situated inlower half of stipe as a tightly fitting band around stipe, darkred-purple, with whitish upper and lower edges, conspicu-ous. Context in pileus white, in stipe cortex dull red-violetand white around central cavity. Odor indistinct.

Basidiospores [20,1,1] 4.6–6.0 3 2.8–3.4 mm, avl 3 avw 5

5.2 3 3.1 mm, Q 5 1.4–1.9, avQ 5 1.7, in side view ellipsoidto oblong, with flattened or convex adaxial side, in frontalview ellipsoid to oblong, slightly thick-walled, smooth,without germ pore, clustering in groups, slightly and slowlystaining in Congo red, nondextrinoid (also after 2 h inMelzer’s reagent), with a pink inner wall in Cresyl blue, withtwo nuclei. Basidia 23–28 3 6.0–7.5 mm, 4-spored, withclamp connection. Lamella edge sterile. Cheilocystidia 17–35(–43) 3 6.5–8.5 mm, narrowly cylindrical, narrowly utri-form, hyaline, slightly thick-walled. Pleurocystidia absent.Pileus covering hymeniderm made up of 30–65 3 9–15 mmclavate to broadly clavate cells, often with long pedicels, withbasal clamp connection; cells moderately tightly knittogether, with brown intracellular pigment. Clamp connec-tions present in all tissues.

Habitat and distribution. Gregarious in smallgroups, terrestrial on irrigated soil, in a garden bedwith ornamental ivy under Sequoia sempervirens (D.Don) Endl. and Olea europaea L., in urban setting.August.

Collection examined. USA. California: Santa Clara Coun-ty, Santa Clara, 28 Aug 2008, J. Brown s.n.

This is the first find of Lepiota lilacea west of theRocky Mountains. Lepiota lilacea is easy to recognizebecause of its distinct annulus in combination withthe pileus covering. There is considerable colorvariation within this species; the pileus and annuluscan be very light to very dark, and lilac tinges might ormight not be apparent. California specimens areslightly different from the European finds in thenarrow cheilocystidia, but all other characters are inconcordance with the European descriptions (Bon1993, Vellinga 2001c).

It is a widespread, weedy species, occurring inurban areas in North America, although not oftenrecorded. Singer (1986) collected it in New York, andfrom my own experience I know that it is common inAnn Arbor, Michigan, and Denver, Colorado, where itfruits in gardens, parks, etc. in August. It also hasbeen found in Minnesota (Michigan fungal herbari-um Website http://www.herb.lsa.umich.edu/cgi-win/

axor32.exe/axorion5parms.ini), and in Iowa (Healypers comm).

Lepiota sublilacea Peck differs particularly in theevanescent annulus (Peck 1897); its pileipellis type isunknown.

4. Lepiota luteophylla Sundb., Mycologia 63:79. 1971.FIG. 4

; Cystolepiota luteophylla (Sundb.) Knudsen, BotTidsskr 73:134. 1978.

Selected description. Sundberg, Mycologia 63:79–81.1971.Lepiota luteophylla is characterized by a dark brown

pileus, yellow ventricose lamellae, an evanescentyellow annulus and the absence of cheilocystidia.The morphological differences with L. neophana arethe darker and more intense colors of the basidio-carps; both species have ventricose lamellae and lackcheilocystidia. The nrITS sequences are quite differ-ent (FIG. 1). Illustrations of the microscopic detailstaken from the type collection are given here (FIG. 4).The spores are slightly metachromatic in Cresyl blueand not dextrinoid. Unfortunately the number ofnuclei could not be assessed.

Knudsen (1978) placed L. luteophylla in genusCystolepiota Singer, although globose cells are absentfrom the pileus covering. The separate cells of thepileus covering are difficult to see, so he might havebeen misled by a section taken parallel to the pileuscovering.

Despite many forays at the type locality, a grove ofCallitropsis macrocarpa south of San Francisco, I havenever found this species myself. It also is known fromMichigan (Smith et al. 1979).

Collection examined. USA. California: San Mateo County,San Francisco watershed, 10 Feb 1970, H.D. Thiers HDT24760 (holotype, SFSU).

5. Lepiota neophana Morgan, J Mycol 12:248. 1906.FIG. 5

5 Lepiota neophana var. europaea Bizio & Migl., BollAss micol ecol Romana 9(27):45. 1993.

FIG. 4. Lepiota luteophylla. A. Spores. B. Basidia. C.Pileus covering elements. All from the holotype (HDT24760). Bar 5 10 mm.

668 MYCOLOGIA

5 Lepiota neophana var. europaea f. papillata Migl. &Perrone, Boll Ass micol ecol Romana 9(27):45.1993.

Excluded. Lepiota neophana sensu Smith, Lloydia17:313–314. 1954 (5 Lepiota sp.); Lepiota neophanasensu Sundberg, Mycotaxon 34:245–247. 1989 (5

Lepiota phaeoderma).Selected description. Bizio et al., Riv Micol 36:228–234.

1993 (as var. europaea and f. papillata).Pileus 10–38 mm, convex, plano-convex with low or

prominent umbo, wavy campanulate to wavy convex withprominent umbo, orange-brown (Mu. 7.5 YR 4/6, 5/6, 6/6,7/6; 5 YR 4/5, 5 YR 4–5/6, 5 YR 5/8), orange-reddish brown(5 YR 3/4) at center, a bit duller when dry, around centerpaler orange brown, orange-brown (halfway around radius 5YR 7/6, 7.5 YR 7–8/5, 5/6, or 7.5 YR 8/4, 10 YR 8/5) to verypale at utmost margin and there cream-colored (10 YR 8/4), smooth and glabrous or radially wrinkled or venosearound center, with some radial grooves when old, with

slightly uneven margin, and in some specimens with whiteannular remnants clinging to margin. Lamellae, L 5 30–40,l 5 1–5, crowded, moderately crowded, free and close tostipe, ventricose, rarely not ventricose but segmentiform,2.5–6 mm wide, whitish at first and soon cream-colored (10YR 8/3–4) to distinctly yellowish (2.5 Y 8/2), with white orconcolorous even edge. Stipe 27–75 3 2–5 mm, cylindricalor slightly wider at utmost base, pale yellowish brown orcream to cream yellow at apex, and there shiny, slightlydarker downward, paler than 5 YR 5/8, with some pinkishbrown or even vinaceous tinges in old specimens, hollow,with white rhizomorphs at base. Annulus whitish andflaring, without cuff, flimsy, evanescent, rarely as somewhitish cream fibrils on both sides of brown rim, oftenclinging to pileus margin and not at stipe. Context thin,cream to yellowish white and dull in pileus, concolorous tostipe surface in stipe. Odor sweet lepiotoid and astringent,such as the sweet component of L. cristata, with a parsley-soapy component, or like L. cristata without the sweetcomponent.

Basidiospores [115,6,6] 4.1–5.7 3 2.6–3.4 mm, avl 3 avw 5

4.7–4.9 3 2.9–3.2 mm, Q 5 1.35–1.9, avQ 5 1.55–1.65, inside view ellipsoid to oblong, with flattened or convexadaxial side, ellipsoid to oblong in frontal view, slightlythick-walled, smooth, without germ pore, clustering ingroups, not colored to pale pink in Congo red, with pinkinner wall in Cresyl blue, nondextrinoid (also after 1 h inMelzer’s reagent), with two nuclei. Basidia 18–30 3 5.0–8.0 mm, 4-spored, with clamp connection. Lamella edgefertile. Cheilocystidia absent. Pleurocystidia absent. Pileuscovering a tightly bound hymeniderm, made up of cells, 23–65 310–25 mm, clavate, broadly clavate, to sphaeropedun-culate, with short to long pedicel, not obviously pigmented.Clamp connections present, especially in hymenium.

Habitat and distribution. Solitary to gregarious insmall groups, terrestrial in duff, known from Calli-tropsis macrocarpa stands, Sequoia sempervirens andmixed forests in coastal zone of California, November–December. Distribution outside California unknown.

Collections examined. USA. California: Humboldt Coun-ty, Humboldt Redwoods State Park, south of Weott, 8 Nov2004, E.C. Vellinga 3228, nrITS GQ203813; Marin County,Mount Tamalpais, 8 Dec 2000, leg. D.E. Desjardin (E.C.Vellinga 2568); Mount Tamalpais, along Bolinas-Fairfaxroad, 15 Nov 2005; near Alpine Lake, 2 Dec 2007, leg. A.Simonin (E.C. Vellinga 3735), nrITS GQ203809, EF1-aGQ375551; Samuel P. Taylor State Park, 28 Nov 2001, E.C.Vellinga 2763; San Mateo County, San Bruno, SkylineCollege, 13 Sept 2009, leg. J.F. Kerekes (coll. E.C. Vellinga4087); San Francisco watershed, 10 Dec 1999, E.C. Vellinga2396; ibidem, 8 Dec 2000, E.C. Vellinga 2602, nrITSAY176492; ibidem, 23 Dec 2002, E.C. Vellinga 2970; ibidem,7 Jan 2003, E.C. Vellinga 2999; ibidem, 28 Jan. 2003, E. C.Vellinga 3011; ibidem, 5 Dec 2003, E.C. Vellinga 3147;ibidem, 1 Dec 2006; ibidem, 25 Nov 2008, E.C. Vellinga 3947,nr ITS GQ203812, EF1-a GQ375550; ibidem, 5 Dec 2008,E.C. Vellinga 3955, nrITS GQ203811, EF1-a GQ375552.USA. Iowa: Story County, Ames, YMCA woods, western part,25 Aug 2006, R. Healy rh59 (ISC); Story County, Ames,

FIG. 5. Lepiota neophana. A. Habit (from ecv3955). B.Habit (from ecv2999). C. Spores. D. Basidia. E. Pileuscovering (microscopic details from ecv2970). Bars: A, B 5

10 mm, C–E 5 10 mm.

VELLINGA: LEPIOTA IN CALIFORNIA 669

YMCA woods, eastern part, 27 Aug 2007, R. Healy rh24nrITS GQ375546, EF1-a GQ375555 (ISC); Webster County,Woodman Hollow State Preserve, 11 Aug 2007, R. Healyrh18 EF1-a GQ375554 and rh39 nrITS GQ375547, EF1-aGQ375553 (ISC).

Lepiota neophana is easy to recognize in the fieldbecause of its brown to pale brown strongly umbo-nate, often slightly radially wrinkled pileus with a lightmargin, which does not break open into smaller scalesand patches, the broadly ventricose yellowish whitelamellae and a whitish annulus on the smooth stipe.Microscopically it is characterized by the smallbinucleate spores, the absence of cheilocystidia andthe tightly knit hymenidermal pileus covering. Lepiotaneophana in the present concept is widely distributed,although rarely recorded.

The species and its name have a long andconvoluted history. Morgan (1906) described L.neophana from Preston (now New Haven), HamiltonCounty, Ohio, as, ‘‘Pileus fleshy, ovoid then campan-ulate and expanded, subumbonate, the flesh thin,firm, white; the dermis thin, tough, the surfacesmooth and glabrous, buff to pale umber, dark brownin the center, the cuticle continuous or at maturitysometimes cracking into irregular areolae. Stipeslender, subequal, tough, fistulous, white above theannulus, pale umber below, with a white-fibrillosecuticle. Lamellae broad, close, white, obtuse behind,free, approximate; spore oblong, obliquely apiculate,4–5 3 3 mic.

Growing on the ground in woods. Preston, O.Pileus 2–3 cm in diameter; the stipe 3–4 cm long and2–3 mm thick. The peculiarity of the plant is itstoughness in all parts, its subcoreaceous texture.’’

The California specimens examined by the presentauthor fit this description very well, although ‘‘tough-ness in all parts’’ and ‘‘subcoreaceous textures’’ arenot characters specifically observed. Murrill (1914)and Kauffman (1924) included the species in theirkeys but added nothing to Morgan’s description;Murrill (1914) said that it was known only from thetype locality.

Smith (1966) studied the type collection andnoted, ‘‘Spores 4–5.5 3 2.2–2.8 mm, cylindric tobroadly ellipsoid in face view and inequilateral inprofile view, wall smooth, not thickened, sporesadhering together in mounts in Melzer’s sol.; verypale rusty brown to unstained; basidia 13–16 3 4.4–5 mm, 4-spored, hyaline in KOH; pleurocystidialacking; cheilocystidia lacking, the gill edges fertile;gill trama very compact, interwoven to subparallel,hyaline in KOH, very pale in Melzer’s sol.; pileustrama floccose, hyaline in KOH, very pale yellow inMelzer’s sol.; cuticle a hymeniform layer of pyriformcells 15–26 3 5–10 mm, hyaline in KOH, scarcely

staining in Melzer’s sol., the surface of the pileus notsmooth but with frequent elevations and depressions,ruptured in some places and the cuticle missing overpart of the surface; clamp connections lacking.’’

Franco-Molano also studied the type collection andnoted that spores are ‘‘inamyloid, ellipsoid tocylindric, (3.6)4.5–6.3 3 2.7–3.6 mm, Q 5 1.52,’’clamp connections are present (New York BotanicalGarden’s Virtual Herbarium http://sweetgum.nybg.org/vh/specimen.php?irn5816627). The sizes of themicroscopic characters provided by Smith (1966) aresmaller than those found in the California specimens,but the spore sizes provided by Franco-Molano agreemuch better.

However Lepiota neophana has been interpreted inseveral other ways. Smith (1954) reported a speciesfrom Michigan under this name. The pileus isdescribed as, ‘‘Pileus 1–3 cm. broad, obtuse to almostglobose when young, becoming broadly conic toalmost plane, with or without a slight umbo, cuticlecontinuous at first, remaining smooth on disc,sometimes splitting radially on the margin andshowing the white flesh, not truly scaly but the cuticlesoon rupturing into small irregular areolae orfurfuraceous patches at least toward the margin, discnear army brown, remainder wood brown or having afaint vinaceous tint or ‘avellaneous’.’’ The Michigancollection is described microscopically with basidium-like cheilocystidia and faintly dextrinoid spores;clamp connections are said to be absent. Neitherthe pileus covering rupturing into small areolae northe basidium-like cheilocystidia fit the original de-scription of L. neophana.

A different interpretation of Lepiota neophana ispresented by Sundberg (1989) from California. Hedescribed a dark almost black species with a pileuscovering that cracks open; cheilocystidia and clampconnections are present and numerous. Here againcharacters of the pileus covering, both color andstructure, do not fit the original description. Thepresence of cheilocystidia is another deviating char-acter. This taxon is described in this paper as the newspecies L. phaeoderma.

Italian authors described a new variety and a newforma of Lepiota neophana, both of which are in goodagreement with the description by Morgan, althoughin general their specimens are paler (Bizio, Migliozziand Zecchin 1993; type study by Vellinga and Huijser1999). The California specimens presented here fittheir descriptions of the species very well, and this wasa decisive factor in considering the name L. neophanafor these collections. The umbonate pileus with thenarrow light margin, the absence of cheilocystidia,and spore characters, all fit very well. Unfortunatelyno molecular data back this up.

670 MYCOLOGIA

Recent collections from northern Iowa (see collec-tions examined) have the same morphologicalcharacters as the ones from California but differslightly in nrITS and more significantly and consis-tently in EF1-a sequences (FIGS. 1, 6). For the timebeing I refrain from describing the Californiaspecimens as a separate taxon; there is no reason todo so on morphological grounds, and to describe anew taxon on molecular data alone seems premature.Sampling is limited and geographically restricted toonly two regions (California and Iowa). The existenceof L. neophana var. europaea might appear to justifydescribing the California specimens as anothervariety. However there is no accepted basis forerecting varietal rank on sequence data.

It might be confusing that the name Lepiotaneophana is now used for a different species thanthe one Sundberg (1989) applied it to, especiallybecause both species co-occur in the same area.Unfortunately the age of the type collection (morethan 100 y) rules out any easy use of molecular data.Furthermore there are conflicting interpretations ofthe species as shown and the likelihood that a speciesfound in Ohio also grows in California might notseem great. Nonetheless the original description, thetype studies, the European interpretation of this

species and last but not least modern collectionsfrom the same general area as where the type wascollected support my identification.

Several species look similar to L. neophana or mightotherwise be confused with it. Lepiota thiersii has apileus covering that splits open into small patches andscales but is microscopically similar to L. neophana.Lepiota luteophylla is also similar to L. neophana buthas a dark brown pileus and deep yellow lamellae.Lepiota subneophana Murrill, described from Florida(Murill 1943), despite its name is a Leucoagaricusspecies with a silky pileus surface with metachromaticspores and without clamp connections (Akers 1997).

Another species also described by Morgan is closeto L. neophana and has its own problems. Morgan(1906) described Lepiota rufipes Morgan as, ‘‘Pileus alittle fleshy, convex, smooth and glabrous, white.Stipe slender, smooth and glabrous, rufescent, palerat the summit; the annulus evanescent. Lamellaebroad, close, white, free, approximate; spores oblong,4–5 3 3 mm. Growing on the ground in woods amongold leaves; Preston, O. Pileus about a centimeter indiameter, the stipe 2–3 cm long.’’

This description could apply to a specimen ofCystolepiota seminuda (Lasch) Bon after the globosecells have been washed off by rain, but the Europeaninterpretation of Lepiota rufipes is a species with ahymeniform pileus covering within Lepiota proper(Kuhner and Maire 1937). In this interpretation thespecies has a pale, cream to pinkish pileus that doesnot break open into scales, no annulus, smalluninucleate spores, and narrowly clavate cheilocysti-dia (Vellinga and Huijser 1999). This taxon is one ofthe few Lepiota species in which the second nucleus ofthe developing spores migrates back to the base of thebasidia instead of staying put (Kuhner 1945). It shareswith L. neophana the tightly knit pileus covering butdiffers in the presence of cheilocystidia and uninu-cleate spores. Lepiota rufipes features in the accountof the Michigan species by Smith (1954) as a rarespecies with a white, almost chalky pileus becomingsordid with age. The rest of the characters are inconcordance with those reported by the Europeanauthors. Smith (1966) did not include L. rufipes inher studies on North American type collections.Fortunately the original Morgan collection of L.rufipes was located in the ISC herbarium with ‘‘type’’written in pencil on the envelope. Little remained ofthe original basidiocarp, but these characters couldbe noted (FIG. 7): basidiospores [20,1,1] in side viewand frontal view 4.1–4.9 3 2.4–2.8 mm, avl 3 avw 5 4.63 2.5 mm, Q 5 1.65–2.05, avQ 5 1.85, oblong tosubcylindrical, smooth, without germ pore, notcongophilous, not dextrinoid, slightly metachromaticin Cresyl blue; basidia 4-spored; cystidia not observed,

FIG. 6. The one most parsimonious tree, based on adataset of EF-1a sequences of L. neophana from Californiaand Iowa, L. phaeoderma, and with L. magnispora and L. cfpseudohelveola as outgroups. One hundred three characterswere parsimony informative. Numbers above branchesindicate branch length, numbers in italics below branchesindicate percentage bootstrap support, based on a fullheuristic search with 10 000 replicates and groups with .

70% support retained.

VELLINGA: LEPIOTA IN CALIFORNIA 671

partly due to the state of the dried collection; lamellatrama regular; pileus covering an epithelium made upof noncolored globose to subglobose, rarely sphaer-opedunculate elements, 17–40 mm diam; clampconnections present.

The relatively narrow spores and the epithelioidpileus covering support synonymy with Cystolepiotaseminuda (Lasch) Bon, as the description alreadysuggested. Accordingly the Eurasian species, com-monly referred to as L. rufipes, must be described asnew.

6. Lepiota phaeoderma Vellinga, sp. nov. FIG. 8MycoBank MB 514125Misapplied. Lepiota neophana sensu Sundberg, Myco-

taxon 34:245–247. 1989.Selected description. Sundberg, Mycotaxon 34:245–247.

1989 (as L. neophana).Lepiotae hymenodermati similis pileo coloribus atris

stipitis basi sine squamulis differt.Holotypus ‘‘USA. California: San Mateo County, San

Francisco watershed, 10 Feb 1970, H.D. Thiers HDT24773.’’ (HOLOTYPE designated here, SFSU)

Etymology. The epithet phaeoderma is a transliter-ation and latinization of Greek words waioz, meaningdark, and derma, skin, because of the dark pileuscovering.

Pileus 22–26 mm, broadly campanulate with broad umbowhen young, later wavy plano-convex with broad umbo,almost black at center (10 YR 3–2/1), and smooth, aroundcenter breaking open into very small dark brown (10 YR 3/3), smooth patches on white to whitish background, verypale at margin; margin just exceeding lamellae. Lamellae, L5 45–50, l 5 1–3, rather crowded to moderately crowded,free and very close to stipe, slightly ventricose up to 3.5 mmwide, rounded near stipe, whitish, cream-colored withslightly pinkish sheen; edge white and cystidiose. Stipe 32–45 3 3.5–5.5 mm, cylindrical, cream at apex, lower down

with whitish fibrillose, when young with slightly fluffycovering over pink-brown or pink-red background, laterwithout any ornamentation or annulus, hollow, with whitemycelial cords at base. Annulus absent. Context white andrather thick in pileus, cream-colored in stipe apex, pinkishbrown toward base. Odor strong, astringent, like the rubberodor of Lepiota cristata, or in addition fruity.

Basidiospores [68,4,4] 4.4–6.3 3 2.5–3.4 mm, avl 3 avw 5

4.9–5.2 3 2.9–3.0 mm, Q 5 1.5–2.1, avQ 5 1.65–1.8, in sideview ellipsoid, oblong, a few subcylindrical, with flattened orconvex adaxial side, in frontal view ellipsoid to subcylind-rical, slightly thick-walled, smooth, without germ pore,clustering in groups, vaguely reacting in Congo red, notdextrinoid (also after 1 h in Melzer’s reagent), with pinkinner wall in Cresyl blue, with two nuclei. Basidia 18–29 3

5.5–7.5 mm, 4-spored, with basal clamp connection. Lamellaedge sterile. Cheilocystidia 18–42 3 5.5–13 mm, clavate tonarrowly clavate, some with long pedicel, hyaline andwithout contents. Pleurocystidia absent. Pileus covering ahymeniderm made up of 23–60 3 10–16 mm, clavate,narrowly clavate to broadly clavate, sphaeropedunculate,with pedicel of varying length, with brown intracellularpigment. Clamp connections present in all tissues.

Habitat and distribution. Solitary or in small groups,terrestrial and saprotrophic in duff of Callitropsismacrocarpa in C. macrocarpa stands and in mixed

FIG. 7. Lepiota rufipes. A. Spores; bar 5 5 mm. B.Elements of pileus covering; bar 5 10 mm. All from theholotype (Morgan s.n.).

FIG. 8. Lepiota phaeoderma. A. Habit (from ecv3000). B.Spores. C. Basidia. D. Cheilocystidia. E. Pileus covering(microscopic details from ecv3016). Bars: A 5 10 mm, B–E5 10 mm.

672 MYCOLOGIA

woodlands. Nov–Apr. Known from San Mateo, SantaClara and Butte counties, California, USA

Collections examined. USA. California: Santa Clara Coun-ty, Palo Alto, Los Tranco Preserve, 25 Nov 2006, D. Smith;California, San Mateo County, San Francisco watershed, 3Jan 1967, H.D. Thiers (Sundberg 1093) (SFSU); ibidem, 16Apr 1967, H.D. Thiers HDT 18829 (SFSU); ibidem, 10 Feb1970, H.D. Thiers HDT 24773 (Holotype, SFSU); ibidem, 7Jan 2003, E.C. Vellinga 3000, nrITS GQ203810, EF1-aGQ375549; ibidem, 28 Jan 2003, E.C. Vellinga 3016.

Lepiota phaeoderma is easily recognized because ofthe dark pileus with small patches around the center,the hymeniform pileus covering and the narrowlyclavate to clavate cheilocystidia. The stipe has noscales and does not have an annulus.

This is Lepiota neophana Morgan sensu Sundberg(1989), which deviates from the original descriptionby Morgan (1906) in the colors of the pileus and thesurface that is cracking open, forming small patchesaround the umbo. A comprehensive discussion of L.neophana is given above.

Sundberg (1989) gave a thorough description of L.phaeoderma (as L. neophana). The above macroscopicdescription is based on my own observations of alimited number of collections of this species; thedescription of the microscopic characters also is basedon some of the collections studied by Sundberg(1989).

7. Lepiota scaberula Vellinga, Mycotaxon 80:290. 2001.Selected description.Vellinga, Mycotaxon 80:290–292.

2001.Macroscopically L. scaberula and its European

counterparts L. cystophoroides Joss. & Riousset andC. cystophora (Malencon) Bon are quite differentfrom the other species treated here, with a muchmore fibrillose-squamose pileus covering and adistinctly fibrillose-woolly stipe. Microscopically thepileus covering is more cutis-like with clavate toglobose upright terminal elements. The more char-acteristic members of this group have a pileuscovering that is either not split up or split up intosmall patches composed of tightly packed narrowlyclavate to clavate cells and a stipe without scales orwith a few scales at the base.

Lepiota scaberula is known from a few places incoastal central California.

8. Lepiota thiersii Sundb., Mycotaxon 34:242. 1989.Selected description.Sundberg, Mycotaxon 34:242–244.

1989.Lepiota thiersii has a pileus covering that splits open

into small patches around a closed disk, a smallannulus, and ellipsoid spores; it lacks cheilocystidia.In the field it can be confused with L. castaneidisca

but differs in the ellipsoid spores and the presence ofcheilocystidia. It differs from L. neophana in thecovering that splits open but shares the small ellipsoidspores and the absence of cheilocystidia with thatspecies. The spores show a pink inner wall in Cresylblue, as do many other species in the group. It differsfrom L. phaeoderma in the much paler, brown pileuscovering, and the absence of cheilocystidia. Theinterpretation of L. neophana by Smith (1954)actually might refer to this species. Lepiota thiersiioccurs in coastal central California but is rarelyencountered or more likely rarely recognized.

KEY TO CALIFORNIAN LEPIOTA SPECIES WITH A

HYMENIFORM PILEUS COVERING

1. Basidiospores bullet-shaped with a truncate base . . 21. Basidiospores ellipsoid. . . . . . . . . . . . . . . . . . . . . . 3

2. Pileus rounded, convex, with red-brown tingedcovering . . . . . . . . . . . . . . . . . 1. L. castaneidisca

2. Pileus in general broadly umbonate withorange-tinged brown covering . . . . . 2. L. cristata

3. Pileus surface not breaking open into scales. . . . . . 43. Pileus surface breaking open, especially around

umbo into patches or scales; cheilocystidia presentor absent (in one species) . . . . . . . . . . . . . . . . . . . 64. Cheilocystidia present and narrowly clavate to

cylindrical . . . L. rufipes sensu European authors(see notes under L. neophana)

4. Cheilocystidia absent . . . . . . . . . . . . . . . . . . . . 55. Pileus dark brown; lamellae intense yellow; annulus

appressed to stipe in young specimens, evanes-cent . . . . . . . . . . . . . . . . . . . . . . . . 4. L. luteophylla

5. Pileus medium to pale brown; lamellae off whiteto pale yellow; annulus present, flaring andwhite fibrillose, sometimes adhering to pileusmargin. . . . . . . . . . . . . . . . . . . . . . . . . . 5. L. neophana6. Cheilocystidia absent . . . . . . . . . . . . 8. L. thiersii6. Cheilocystidia present . . . . . . . . . . . . . . . . . . . 7

7. Annulus present, conspicuous, with dark cover-ing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3. L. lilacea

7. Annulus absent . . . . . . . . . . . . . . . . . . . . . . . . . . . 88. Pileus surface very dark brown, breaking up

into small flat patches around umbo; stipesmooth . . . . . . . . . . . . . . . . . . . 6. L. phaeoderma

8. Pileus surface cream to yellow brown, irregu-larly fibrillose-squamose around umbo; stipewhitish-woolly in lower half . . . . . 7. L. scaberula

ACKNOWLEDGMENTS

The San Francisco Public Utilities Commission grantedpermission to inventory the Monterey cypress groves of theSan Francisco watershed area (west of Crystal SpringReservoir). Dimitar Bojantchev, John Brown, JenniferKerekes, Anna Simonin, Douglas Smith and the curatorsof the herbaria ISC, NY and SFSU contributed material forstudy and examination. Dr Nicholas P. Money and Dr

VELLINGA: LEPIOTA IN CALIFORNIA 673

Michael A. Vincent were extremely helpful in tracing thelocation of A.P. Morgan’s ‘‘Preston’’. Rosanne Healy andDeborah Q. Lewis of the Ada Hayden Herbarium (ISC) sentme original Morgan material of L. rufipes, and moderncollections of L. neophana from Iowa. Dr Jan FritsVeldkamp proposed the Latin diagnosis. John Lennie madevery helpful suggestions regarding presentation. Remarksand comments by the two anonymous reviewers and Dr P.B.Matheny were very useful. Financial support through NSFgrant DEB 0618293 is gratefully acknowledged.

LITERATURE CITED

Akers BP. 1997. The family Lepiotaceae (Agaricales,Basidiomycetes) in Florida [Doctoral dissertation].Carbondale: Southern Illinois Univ. 253 p.

Bizio E, Migliozzi V, Zecchin G. 1993. La sezione Integrellae(Kuhner ex M. Bon) M. Bon del genere Lepiota(Persoon) Gray. Riv Micol 36:223–244.

Bon M. 1993. Flore mycologique d’Europe 3. Les Lepiotes.Doc mycol Memoire hors serie 3:1–153. LepiotaceaeRoze.

Gardes M, Bruns TD. 1993. ITS primers with enhancedspecificity for basidiomycetes—application to the iden-tification of mycorrhizae and rusts. Mol Ecol 2:113–118.

Holmgren PK, Holmgren NH. 1998 [continuously updat-ed]. Index Herbariorum: a global directory of publicherbaria and associated staff. New York BotanicalGarden’s Virtual Herbarium. http://sweetgum.nybg.org/ih/ [accessed 13 Jul 2009].

Kauffman CH. 1924. The genus Lepiota in the United States.Papers Mich Acad Sci, Arts Lett 4:319–344.

Knudsen H. 1978. Notes on Cystolepiota Sing. and LepiotaS.F. Gray. Bot Tidsskr 73:124–136.

Kropp BR. 2008. Dermoloma inconspicuum from Belize withmolecular support for its placement in the Agaricaceae.Mycotaxon 104:235–240.

Kuhner R. 1945. Nouvelles recherches sur les divisionsnucleaires dans la baside et les spores des Agaricales.Cr Acad Sci Paris 220:618–620.

———, Maire R. 1937. Trois Lepiotes peu connues. Bull SocHist nat Afr N 28:108–112.

Larkin MA, Blackshields G, Brown NP, Chenna R, McGetti-gan PA, McWilliam H, Valentin F, Wallace IM, Wilm A,Lopez R, Thompson JD, Gibson TJ, Higgins DG. 2007.Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948.

Lee C, Grasso C, Sharlow M. 2002. Multiple sequencealignment using partial order graphs. Bioinformatics18:452–464.

Liang JF, Xu J, Yang ZL. 2009. Divergence, dispersal andrecombination in Lepiota cristata from China. FungalDiversity 38:105–124.

Morgan AP. 1906. North American species of Lepiota. JMycol 12:154–159; 195–203; 242–248.

Murrill WA. 1912. The Agaricaceae of the Pacific Coast II.Mycologia 4:231–262.

———. 1914. Agaricaceae (pars). N Am Flora 10(1):1–65.———. 1943. More new fungi from Florida. Lloydia 6:207–

228.1975. MunsellTM soil color charts. Baltimore.Peck CH. 1897. New species of fungi. Bull Torrey Bot Club

24:137–147.Rehner SA, Buckley E. 2005. A Beauveria phylogeny

inferred from nuclear ITS and EF1- a sequences:evidence for cryptic diversification and links toCordyceps teleomorphs. Mycologia 97:84–98.

Singer R. 1986. The Agaricales in modern taxonomy. 4thed. Koenigstein: Koeltz Scientific Books. 981 p.

Smith AH, Smith HV, Weber NS. 1979. How to know thegilled mushrooms. Dubuque, Iowa: W.C. Brown Co.334 p.

Smith HV. 1954. A revision of the Michigan species ofLepiota. Lloydia 17:307–328.

———. 1966. Contributions toward a monograph on thegenus Lepiota I. Type studies in the genus Lepiota.Mycopathol Mycol Appl 29:97–117.

Sundberg WJ. 1967. The family Lepiotaceae in California[Master’s thesis]. San Francisco: San Francisco StateUniv Press. 219 p.

———. 1971. A new species of Lepiota. Mycologia 63:79–82.———. 1989. Lepiota sensu lato in California III. Species

with a hymeniform pileipellis. Mycotaxon 34:239–248.Swofford DL. 2002. PAUP*: phylogenetic analysis using

parsimony (*and other methods). Version 4. Sunder-land, Massachusetts: Sinauer Associates.

Vellinga EC. 2001a. Studies in Lepiota III. Some speciesfrom California. USA. Mycotaxon 80:285–296.

———. 2001b. Studies in Lepiota IV. Lepiota cristata and L.castaneidisca. Mycotaxon 80:297–306.

———. 2001c. Lepiota. In: Noordeloos ME, Kuyper ThW,Vellinga EC, eds. Flora agaricina neerlandica 5:109–151.

———. 2003. Phylogeny of Lepiota (Agaricaceae). Evidencefrom nrITS and nrLSU sequences. Mycol Prog 2:305–322.

———. 2004a. Ecology and distribution of lepiotaceousfungi (Agaricaceae). Nova Hedwig 78:273–299.

———. 2004b. Genera in the family Agaricaceae—evidencefrom nrITS and nrLSU sequences. Mycol Res 108:354–377.

———. 2007. Lepiotaceous fungi in California, USA 2.Lepiota rhodophylla. Mycotaxon 98:205–211.

———, Huijser HA. (1998) 1999. Studies in Lepiota I.Species with a hymeniform pileus covering. Belg J Bot131:191–210.

———, Noordeloos ME. 2001. Glossary. In: NoordeloosME, Kuyper ThW, Vellinga EC, eds. Flora agaricinaneerlandica 5:6–11.

Zeller SM. 1933. New or noteworthy agarics from Oregon.Mycologia 25:376–391.

674 MYCOLOGIA