Amplistroma Gen. Nov. and Its Relation to Walirothiella, Two Genera with Globose Ascospores and Acrodontium-Like Anamorphs

M't'cologia, 101(6), 2009, pp. 904-919. DOl; 10.3852/08-213 r 2009 by The Mycological Society, of America, Lawrcnce, KS 66044-8897

Amplistroma gen. nov. and its relation to Walirothiella, two genera with globose ascospores and acrodontium-like anamorphs

Sabine M. Huhnclorf' INTRODUCTION Botany Department, Field Museum of Natural history Genus Walirot/iieiia Sacc. recently has been rede- Chicago, Illinois 60605-2496 scribed and the type species, W. congregata (Wallr.) Andrew N. Miller Sacc., was neotypifIed based on collections from Illinois Natural Histoiy Survey, University of Illinois at France and Ukraine (Réhlová and Seifert 2004). Urbana-Champaign, Champaign, Illinois 61820-6970 The genus is distinct, in its globose, long-necked Matthew Greif asconiata, its wide, long, tapering paraphyscs and its Botany Department, Field Museum of Natural History, cylindrical, stipitate asci with eight, small, globose Chicago, Illinois 60605-2496 ascospores. Surveys of wood-inhabiting Sordariomy- cetes in Puerto Rico and Great Smoky Mountains Gary J• Samuels National Park in the eastern United States uncovered USDA-ARS, Systematic MycoloTy & Microbiology several specimens that match the description of lV Laboratory, Room 304, B-01 IA, 10300 Baltimore Avenue, Beltsville, Maryland 20705-2350 con gregala, and a collection from Puerto Rico was obtained in culture. A few years earlier several specimens that shared key characteristics of W. were conveyed to us. These specimens Abstract: Ainpiislroma is described as a new genus congregala have the same distinctive eight, globose-spored asci for A. caroiinianu,n, A. dirninulisporum, A. guianense, and wide parapityses that are long and tapering above A. hallini,rii, A. ravum, A. iartareum and A. xyiar- the asci. They differ however in having ascomata ioides. Species of Ampiistroma are distinguished by stromata. To reveal the large stromata of textura in tricata with polystichous united in large, flesh y co'ngregata and these unknown ascomata and long necks that are either erumpcnt relationships of V fungi new sequences of the LSU were analyzed and from the stromatal surface or form bumps or compared to known datasets with maximum parsinlo- protuberances. The type collection of Ceralostoma n and Bayesian analyses. .cphaerospermum was examined and found to be y SflOOUS with Wallrol.hiella cangregata. The distribution of W congregata is expanded by collections MATERIALS AND METI I01)S from Costa Rica, the eastern United States and /axon sainpilng.—laxa sequenced iii this suidy are listed Walliothielia congregala has ascomata Puerto Rico. (T. tut'. t) with collection data provided under the exam- that are long-necked and develop individually or are ined specimens for each taxon. Representatives from gregarious on the substrate but do not form large families and orders within the Sordariomycetes were stromata. Aniplistroma and Waiirot/tiella are distin- in i-It ided to determine the phvlogenetic position of, the guished by small asci with eight, minute, globose target taxa. In addition we also included sequences br two ascospores. An acrodonuum-like anamorph occurs in stromatic species, Pachvtrype rimosa F.A. Fernhndez Ct al both genera. Phylogenetic analyses of 28S large- (COSTA RICA. Cartago, Paique Nacional TapantI, on I iii subunit rDNA sequences group these laxa in a well diani tog. 441I-2002, l'.A. Fern(indez FF1066, F) and P. (Penz. & Sacc.) M.E. Barr et al (USA. Hawaii. supported dade distinct from known orders within pr'nceps Manuka Park, on dead wood of Pvlelrosideros pol'i'moipha 2!- the Sordariomycetidae but showing unsupported X1-1998, J.D. R(gers. F). All voucher specimens are relationships with the Chaetosphaeriales and the deposited in the Field Museum Mycology Herbarium (F). Magnaporthaceae. Family Amplistromataceae is de- Ascomata were mounted in water and replaced with Scribed forfor this dade and placed within the lactophenol containing a/tire A. A minimum of 30 asci, Sordariomycetidae incertae sedis. ascoS1)ores and conidia were measured with Scion Image Key words: Arnplistromataceae, Ascomycota, (www.scioricorp.com ), and measurements were made and LSU rDNA, systematics, wood-inhabiting fungi images were captured of material in both mounting fluids. Aseomata were sectioned at 5 pm for light microscopy with the techniques of Huhndorf (1991). Images were captured with photomacrography, bright field (BF), phase contrast (PH) and difterential interference microscopy (DIG), and Accepted for publication 5 May 2009. photographic plates were produced following the methods Cori-esponding author. E-mail: shuhndorfifieidmuseum.oig of Htihndorf and Ferniindez (1998). Methods for scanning

904 HullNl)ORF El' AvlJ'I.I.sI70tL4 GEN. NOV. 905

TAint: I. l'axa sequenced in this study

Taxon Source GenBank accession number

A mpljsfrnma ((10)/i /ti(lflUfl? Ascoinata, Doi Sfl FJ532376

A inp/cs-lroina cam/in/a no ni Culture, BF109923 FJ5%2377

A rnp/it(roina guian(-n.u' Ascomata, G.J.s. 5740 FJ532 380

A nip/is Ironni h (li/ingi? Ascomata, R.E. I-Ia] Iii ig 7389 FJ532379

.4 mpiislroma ravu ill A.scomata, SMH4958 FJ532378

Pa chVti)je pr/n reps Ascontata, Rogers sit FJ532382

I'ac/'(r)'pe rim nsa A.ScoIflata, FF1066 FJ53238 I

lVa//rol/iielia can grega Ia Ascoinata, ANM8 I FJ532374

1Vallrathze//a 'nflgregala Culture, SMH 1760 FJ532375

electron microscopy (SEM) follow Meyer and Plaskowiti for the nucleotide substitution model and for priors with (1989). Cultures were obtained b y germinating ascospores the chain temperature set at 0.2. The nucleotidc substitu- on water agar or cornmeal dextrose agar. Germination tion model was chosen with Modeltesi. 3.7 (Posada and occurred at a high percentage after 48 Ii. Single ascospures Cnimm(all 1998). Every 10011i tree was sampled for 2 500000 and asci with germinating spores were isolated with a generations. The first 6250 trees (25% of total) were ti) icromnanipulalor 01 bN hand. Growt h observations of discarded as burn-in and posterior probabilities ^:95% were niimltispore isolates were made oil cornmeal agar and malt determined front a consensus tree generated with the extract agar at I wk. Colors of the cultures follow Rayner remaining 18750 trees. The consensus tree was viewed with 1970). The cultures are deposited at Centraalhureau voor PAUP 4.0 blO (Swofford 2002). Schinsmelciiltures, Netherlands (CRS)

1)A'A extraction, 1'(Js' (/Fflp/)jicaliafl and sequencing. —De- R1-:S1t:ls tailed protocols for the extraction, amplification and .Sequenre alignment and ph'1ogenelic analfses.—TIIc sequencing of DNA are described in Hithudorl et a] (2008). LSU alignment contained a total of 87 taxa and 1046 Sequence alignment and pl's'fageneIic analyses.—Methods for characters, 534 of which were constant and 404 were the alignment of I .SU sequences arc described in Huhndorf parsimony informative. The heuristic search pro- et al (2008) Sequence data for nine species were aligned duced 74 equally parsimonious trees (one of which with sequences of 78 Species retrieved from GenBaitk, is shown. in FI(;. 1) with a comlsistencv index of ' 0.315 representing major clades within the Sordariomycetes and a retention index of 0.654. where I ,SU data was available, using Se-Al 2.0aI 1 Carbon (Raiubaut 2002). The data matrix was assembled with Se-Al, Species ie/afionsliips.—Ihe LSU phylogenv contains a then exported to Mesquite 1.12 (Madison and Madison dade representing the proposed new family support- 2006) where it was automatically aligned with Cltistal X 1.8,3' cd by both bootstrap support and significan t Bayesian (Thompson Ct al 1 997). The aligned matrix was imported posterior probability. These data support a strong dade hack into Se-Al and edited manually. .Sculeilinia scutetlaia containing aiiraihieila con .,rrgata and the proposed DQ247806 was used as outgmoitp taxon. A maximilul new genus. Another species, parsintonv analysis was perfoinmed with PAUP 4.0 1)10 I/V subzculo.sa, does not (Swofford 2002).A heuristic search consisting of 100 cluster with W. congregata and is of uncertain placement random Stepwise addition replicates was conducted with in these analyses. Om-dinal placement (if the newlamily is parsimony as the optimnality critel-ioil with gaps treated as not supported so it is referred to the Sordariomvcetidae missing data. iree-biscction-reconncction was chosen as the in certae sedis. Pachytiype rtmosaand P. JirIn reps find their branch-swapping algorithm with tct:mRrt:s option in effect. placement in the Diaporthales. Branch support was estimated by performing 1000 bootstrap replicates with a heuristic search consisting of' 100 TAXONOMY random stepwise addition replicates for each bootstrap replicate with the above settings-and the MAXrREF.S setting Ainplistromataceae Huhndorf', AN. Mill., M. Greif & set to 10000 (Felsenstein 1985). The tree was viewed with Samuels, fain. nov. PAUP 4.0 blO (Swofford 2002). A Bayesian analysis was MycoBank MB 513238 conducted with MrBaves 3.1.2 (1-luelsenbeck em al 2001), Stromata supet-ficialis, turbinata vel ohovoidea vel irregu- which approximates posterior probabilities of clades with a Iaritei- pulvinata, lextltt-a niollis vel cartilaginca; vel stromata Markov chain Monte Carlo (MCMC) method (1-luelsenbcck niillus ci pemitliecia sepemata vel aggregata, subiculuni and Ronqimist 2001). The data matrix was analyzed with the liu'pliae sd nullits. Petithecia glohosa vel sithglobosa, general time reversible model of substitution including polysticha vel momiosticha, longimostris. Paraphvsesahun- estimation of invariant sites and assuming a discrete gamma clans. Asci abitmidans, cvlimm(ImiCi vel clavati. .AScosporae distribution (GTR + I + G) with six rate categories provided g]obosae.

906 MYCOLOGIA

100 Wallrothiella congregata FJ532374 Walirothiella congregata FJ532375 Amplistroma guianense FJ532380 100 Amplistro a carolinianum FJ532376 Amplistromataceae Amplistroma caroinianum FJ532377 81 Amplistroma ravum F.1532378 Amplistroma hallingii FJ532379 Melanopsammella gonytrichii AF466085 Zignoella pulviscula AF466091 I Chaetosphaeriales 96 Ceratosphaerella castillensis EU527997 Ceratosphaerella rhizomorpha EU527998 74 Ophioceras commune DQ341 500 ^84 Ophioceras hongkongense DQ341 509 Ophioceras dolichostomum EU528000 100 Ophioceras leptosporum DQ341510 Ophiocerus chiangdaoense EU571 272 Mycoleptodiscus coloratus DQ341 499 MuraeriatfricanaEU527995 Magnaporthaceae 78 Muraeriata cotapsa EU527996 Buergen erula spartinae D0341492 fl f Magnaporthe salvinii D0341498 886 Gaeumannomyces graminis var. graminis D0341 496 4 79 Gaeumannomyces graminis var. tritici 00341497 M agnaporthe grisea AF362554 Dactylaria higginsii 00341512 52 72 Pyricularia borealis DQ34151 1 -y4Ceratosphaeria lampadophora AY761 084 jratosphaeria phialidica AY346295 Pseudohalorsectria suthepensis DQ341513 Pseudohalonectria lignicola AY346299 (1 100 Togninia sp. DQ649065 0 Togninia minima AY761082 Togninia fraxinopennsylvanica AY761 083 0) Togninia novae-zealandiae AY761081 0 55 r Amphiporthe hranicensis D0323521 3 Gnomonia gnomon AF408361 (1 100 7 Cryptodiaporthe aesculi AF408342 (0 Discula destructiva AF362568 66 10 %elanconis stilbostoma AF362567 0)SD 52 Melanconis alni AF362566 gg Schizoparrne botrytidis AF408383 100EndotheIla gyrosa AF362555 100 Chrysoporthe cubensis AF408338 170 Diaporthe arctii AF362562 Diaporthales Diaporthe eres AF408350 99 Valsa anibiens AF362564 Leucostoma niveum AF362558 96 Pachytrype rimosa FJ532381 Pachytrype princeps FJ532382 100 alosphaeria puichella AY761075 Toynirriella acerosa AY761076 Calosphaeriales urostoma ootheca AY761079 100" Chaetomium globosum 61-286403 100Chaetomium datum DQ368628 Sordaria fimicola AY545728 Sordariales Gelasinospora tetrasperma DQ470980 67 90 Cornipulvina ellipsoides DQ231441 Boliniales Camarops ustulinoides AY346267 Wallrothiella subiculosa U17428 Cryptendoxyla hypophloia AF096190 Albertiniella polyporicola AF096185 Albertiniella sp. AY346256 Cephalothecaceae Cephalotheca sulfurea AF431 950 hiostoma stenoceras DQ836904 Ophiostoma piliferum DQ470955 Ophiostomotales endus austriacus AY590293 'f^iatascus triseptatus AY780049 hongkongensis AR 32319 Ann ulatascaceae elasma sordida AY761087 tomella pyrenaica 00076323 t96 itella crinigera AY761086 mitella cirrhosa AY761085 tomitella pallibrunnea E1J527994 entomitella tropica E1J527999 100 Cryptadelphia groenendalensis EU528001 Cryptadeiphia groenendalensis EU528007 Rhamphoria delicatula AF261068 Diatrype disciformis 00470964 6 50 Xylaria acuta AY544676 Xylaria hypoxylon AY544648 Xylariomycetidae Cairria graminis AY431 949 88 Hypocrea citrina AY544649 Bionectria ochroleuca DQ862027 Ambrosiella xylebori 00470979 Hypocreomycetidae Pleospora herbarum DQ678049 Dothidea sambuci AY544681 Dothideomycetes - Scutellinia scutellata 00247806 Pezizomycetes I - 10 changes

Fit;. 1. Phylogenetic flee obtained from a maximum parsimony analysis showing the position of the Ainplistronsataceac and other taxa based on large stibtittit rDNA sequence data. The family forms a strongly supported chide of uncertain position witluri the Sordariomvceiidae. Bootstrap support valises ^50% are shown above branches and Bayesian support ^95% is indicated by thickened branches.

HUHNDORF ET AL: AvipJJsrI?ovrA GEN. NOV. 907

Tvpus familia. A.inpiisrorna Hiihndorf, A.N. Mill., dense, coriaceous, not cartilaginous, stroma M. Greif & Samuels posed of loosely woven, hyaline to pale brown Stromata superficial, turbinate, obovoid to irregu- hyphae of textura intricata; somewhat more densely larly pulvinatc, texture soft or firm; or stromata al)sent woven and slightly darker in color at stroma surface. and perithecia separate to clustered, with hyphal Perithecia subglobose, 300-325 pill immersed, subiculum or absent. Perithecia globose or subglo- polystichous in two or three layers just under stroma bose, polystichous or monostichous, with long necks. surface, with long necks papillate and raised above Paraphyses abundant. A.sci numerous, cylindrical to stroma surface. Perithecial walls composed of pale clavate. Ascospores globose. brown, thin walled, flattened pseudoparenchyma, 15-20 .in1 thick. Paraphyses abundant, ca. 4-6.5 pm Arnplistroma Huhndorf, A.N. Mill., M. Greif & wide just above asci, long tapering to a narrow apex. Satnuels, gen. nov. Asci with spore-bearing part 24-27 X 3.5-4.5 pm, MycoBank MB 513239 With stipe 10-25 pill long. Ascospores hyaline, globose to slightly depressed globose, smooth, Stromata superlicialis, turbinata vel obovoidea vel irr('gLl_ 2.6- laliter pulvinata, textLlra niollis vel cartilaginea. Pcrithccia 3.4 pm diam. Anamorpit acrodontium-like in culture globosa vel subglobosa, polvsticha vcl monosticha, long- and occurring on young stromata and near base of' irostris. Paraphyses ahundans. Asci abtuidans, cvluidrici ye] older stromata. On substrate, colony brown, same clavati. AScoSporae globosae. color as surface of stroma. In culture colony growth Typu.s generic. Amplistroma caroljnianum Huhndorf, 12-16 mm on CMA and 26-28 mm oil after A.N. Mill., M. Greif & Samuels. 7 d. Oil surface hyaline margin with off-white Etymology. L. amplus = ample, large, great, refers to center (light buff 45), appearing powdery from the asconmta structure. abundant aerial, sporulaung conidiophores, surface Stromata superficial, turbinate, obovoid to irregularly hyphae sparse, reverse hyaline to light brown in pulvinate, texture soft or firm. Pcrithccia globose or center of colon y (hyaline-buff 45), no exiidates. On subglobose, polystichous or monostichous, with long MEA surface light brown (light hazel 88), reverse tan necks. Paraphyses abundant. Asci numerous, thin- to light brown (huff 45—honey 64), no exudates, walled, cylindrical to clavate, stipitate, with eight hyphal appearance same as on CMA. Conidiophores ascospores uniseriately arranged. Ascus apical ring arising plagiotropicallv, at times orthotropicallv, minute to inconspicuous, not staining in Melzer's iodine from hyaline to light brown surface hyphae, ascen- reagent. A.scospores globose, appearing thick-walled. dant to suberect, not strongly differentiated, hyaline to light brown, with whorls of secondary and tertiary branching arising plagiotropically along the length. Amplistroma carolinianurn Huhndorf, A.N. Mill., M. Conidiogenous cells arising in verticils or whorls on the Greif & Samuels, sp. nov. Fius. 2-1 7 branches, lageniform to anipulliform, 5.5-7.5 X 2.0- MycoBank MB 513240 3.0 pill, apex forming elongate rachis, proliferating Stromata inegulariter turhinata ye] obovoidea conncClivo sympodially, straight, flexuous, 8-20 X 0.8-1.3 pm, with lato, 4-8 mm tunga X 3-7 mm lata X 2-4 InIn crassa. perpendicular denticulate, blunt, conicliogenous pegs. Superfiries fulva vel umbrina, papillae hrcvitcr et fuscae; intus crerncas. Tcxtura coriacea et firma; hyphae Lextura Conidia light brown, pyriform to ohovoid with a inimicata laxa. Perithecia globosa 300-325 pm diatn, poly- roughened, basal frill, 2,3-2.8 X 1.8-2.3 pm. sticha, collis longis praedita; ostiola papillata conspicua. Habitat. Oil bark or wood. Paraphvses abundans, ca. 4-6.5 pm ad hasim, versus apiceni Known distribution. United States (North Carolina, perangustam. A.sci cylindrici vel clavati, pars sponifir 24-27 Maryland). X 3.5-4.5 pm, stipitati, 10-25 pin longi, octospori, uniser- .Specinens examined: UNITED STATES. Maryland, How- iati. Ascosporae glohosae, 2.6-3.4 pin diam, hyalinae. ard County, Sykesville, 3 mi S, oil North, just past 70 Etymology. Carolinianum refers to the type collec- junction, Patapsco Valley State Park, 21-IX-1999, BE. tion locality. Overton,, BE09923 (F), culture (CBS 124655): North Subiculum restricted to immediate vicinity of Carolina, Macon County, Ellicott Rock Trail, ofT Bull Pen stroma, cream to tan or grayish brown. Stroma Road, 35 301'N, 83°08'W, elev. 3000 feet, on hark, 14-X- irregularly turbinate to obovoid oil connective 1990, Y. Doi, A. Y. Ross,nan, C. J..S'anne1s s.ri., dried culture G.J.S. 90-43 (holotypc BPI 878925, (half to three-quarters width of stroma), about 4- isotype F). 8 turn X 3-7 mm broad X 2-4 mm thick. Stroma surface tan to dull brown, papillate with dark Amplistroma diminutisporum Huhncloif, AN. Mill., brown, short ostiolar necks; older stromata darker M. Greif & Samuels, sp. nov. Fius. 18-23 With surface covered with densely packed darker MycoBank MB 513241 brown necks; interior cream-colored. Texture firm, Stromata illegulariter turbinata vel ohovoidea conncctivo

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HUHNDORF Er AL: AMPLISTROMA GEN. NOV. 909

lato, 7-8 mm longa X 6-7 mm lata X 5-6 mm crassa. gelatinosus matricis. Perithecia subglobosa 250-280 jim Superficies fulva yel umbrina, aspera cum papillae numer- diam, polysticha, collis longis praedita; ostiola papillata. osae breviter et fuscae; intus viridis ubi madefacta, Paraphyses abundans, Ca. 2.5-4.5 jim supra ascos, versus atroviridis ubi sicca. Textura cartilaginea ubi sicca, gum- apicem perangustam. Asci cylindrici vel clavati, pars sporifer mosa ubi madefacta; hyphae textura intricata laxa in 18-25 >< 3.4-5.4 gm, stipitati, 10-14 jim longi, octospori, gelatinosus matricis. Perithecia globosa 250-300 pm diam, uniseriati. Ascosporae globosae, 2.7-3.0 jim diam, hyalinae. polysticha, collis longis praedita; ostiola papillata. Paraph - Etymology. Guianensis refers to the collection locality. yses abundans, Ca. 10.5-13.5 jim supra ascos, versus apicem Subiculum not seen. Stroma lobed, irregularly perangustam. Asci cylindrici vel clavati, pars sporifer 13.5- turbinate to obovoid on wide connective (ca. three- 17.0 >< 2.0-2.6 pm, stipitati, 9-12 gm longi, octospori, quarters width of stroma), in the order of 12-13 mm uniseriati. Ascosporae globosae, 1.7-2.2 pm diam, hyalinae. long X 9-10 mm broad X 5-7 mm thick, based on Etymology. diminutus refers to the small ascospores. dried stromata. Stroma surface tan to dull brown, Subiculum not seen. Stroma irregularly turbinate to papillate with dark brown, short ostiolar necks; obovoid on moderately wide Connective (ca. half interior greenish when fresh and dark when dry. width of stroma), about 7-8 mm long X 6-7 mm Texture rubbery when fresh, cartilaginous when dry, broad X 5-6 mm thick, based on dried stroma, fresh stroma composed of loosely woven, hyaline to pale strornata larger. Stroma surface tan to dull brown, brown hyphae of textura intricata in a gelatinous roughened papillate with dark brown, short ostiolar matrix; hyphae somewhat more densely woven, slightly necks; interior greenish when fresh and dark when darker and not gelatinized at the stroma surface. dry. Texture rubbery when fresh, cartilaginous when Perithecia subglobose, 250-280 jim diam, immersed, dry, stroma composed of loosely woven, hyaline to polystichous in 6-7 layers just under stroma surface, pale brown hyphae of textura intricata in a gelatinous with long necks papillate and protruding above stroma matrix; hyphae somewhat more densely woven, surface, appearing as dark bumps. Perithecial walls slightly darker in color and not gelatinized at stromal composed of pale brown, thin walled, irregular to surface. Perithecia subglobose, 250-300 jim diam, flattened pseudoparenchyrna, scarcely different from immersed, polystichous in 3-4 layers just under the surrounding stroma tissue, Ca. 20-25 jim thick. stroma surface, with long necks papillate and slightly Paraphyses abundant, Ca. 2.5-4.5 gm wide just above raised above stroma surface, appearing as dark the asci, long tapering above. Asci with spore-bearing bumps. Perithecial walls composed of pale brown, part 18-25 X 3.4-5.4 jim with stipe Ca. 10-14 gm long. thin walled, irregular to flattened pseudoparenchy- Ascospores hyaline, globose to slightly depressed ma, scarcely different from surrounding stroma globose, smooth, 2.7-3.0 gm diam. tissue, Ca. 20-25 gm thick. Paraphyses abundant, Ca. Anamrnph. Not found on the stromata. 10.5-13.5 jim wide just above asci, long, tapering Habitat. On dead wood. above. Asci with spore-bearing part 13.5-17.0 >< 2.0- Known distribution. French Guiana, only known 2.6 pm, with stipe Ca. 9-12 pm long. Ascospores from the type collection. hyaline, globose to slightly depressed globose, Specimens examined: FRENCH GUIANA. St-Laurent-du- smooth, 1.7-2.2 jim diam. Maroni, Commune de Maripasoula, Upper Marouini River, Anamorph. Not found on the stromata. 2 km N of Oumanfou-Lange Soula, 02°52'N, 54°00'W, elev. Habitat. On dead wood. 150 m, on dead wood, 12,14-VIII-1987, G.J. Samuels, JJ. Known distribution. French Guiana, known only deGranville, L. Allorge, W Hahn, M. Hoff G.J.S. 5740 from the type collection. (holotype NY; isotype F). Specimens examined: FRENCH GUIANA. St-Laurent-du- Maroni, Commune de Saul, ca. 7 km SW of Saul toward Mount Galbao, elev. 400-450 m, on recently dead tree, 11-I- Amplistroma hallmgii Huhndorf, AN. Mill., M. Greif 1986, G.J. Samuels, JR. Boise, G.J.S. 2758, (holotype NY; & Samuels, sp. nov. FIGS. 31-40 isotype F). MycoBank MB 513243 Stromata globosa depressa vel obampulliforma connectivo attenuato, 30-35 mm longa >< 20-30 mm lata X 20- Amplistroma guianense Huhndorf, A.N. Mill., M. 25 mm crassa. Superficies fulva yel umbrina, aspera cum Greif & Samuels, sp. nov. FIGS. 24-30 papillae numerosae breviter et fuscae; intus viridis ubi MycoBank MB 513242 madefacta, atroviridis ubi sicca. Textura cartilaginea ubi Stromata irregulariter turbinata vel obovoidea connectivo sicca, gummosa ubi madefacta; hyphae hyphae textura lato, 12-13 mm longa X 9-10 mm lata X 5-7 mm crassa. intricata laxa in gelatinosus matricis. Perithecia subglobosa Superficies fulva vel umbrina, aspera cum papillae numer- 225-275 pm diarn, polysticha, collis longis praedita; ostiola osae breviter et fuscae; intus viridis ubi madefacta, papillata. Paraphyses abundans, Ca. 5.0-7.5 pm supra ascos, atroviridis uhi sicca. Textura cartilaginea ubi sicca, gum- versus apicem perangustam. Asci cylindrici yel clayati, pars mosa ubi madefacta; hyphae textura intricata laxa in sporifer 22.5-26.5(-29.5) X 3.5-4.5 jim, stipitati, 17-22 pm

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HUHNI)QRF ET AL: AMPLISTROMA (EN. NOV. 911

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Fics. 31-40. Ainpli,slroma halhngit. 31, 33. Strornata. 32. Stromatal surface. 34. 1araphyses. 35, 37-39. Asci with ascospores. 36. Longitudinal section through Stronla. 40. Longitudinal section through stromal tissue. 31-33 by photomicrography; 37, 38, 40 by I)IC; 34-36, 39 by PH. Bars: 31, 33 = 5 mm; 32 = 1 mm; 36 = 100 tim; 34, 35, 37-40 = 10 pm. All from REH7389.

longi, uctospori, uniseriati. Ascosporae glohosae, 3.0- 20-30 mm broad X 20-25 mm thick based on dried 3.7 MW diaiii. hvalinae. stroma. Stroma surface tan to dull brown, papillate Etrno1ogv. Named in honor of the collector. with dark brown, short ostiolar necks; interior Suhiculurn not seen. Stroma depressed globose to greenish when fresh and dark when dr y. Texture ohampullifbrm on narrow connective tissue (less than rubbery when fresh, cartilaginous when dry, entire one-half width of stroma), about 30-35 mm long x stroma composed of loosely woven, hyaline to pale

¶-) 12 Ni \( :oi.()( ;L\ l)i'O\Vll hvphae of textura intricata in a gelatinous monostichous, with long necks raised above stronla matrix; hvphae somewhat more densel y woven, surface, up to 600 pm long, 70-100 pm wide. Petit hecial slightly darker and 1101 gelatinized at stroina surface. walls composed of brown, thin walled, flattened Perithecia siibglohose, 225-275 pin diam, immersed, pseudopai'enchy'ma, 30-40 pm thick. Pai'aphvses abun- in polvst ichous 4-5 layers just under Si roma surface, dant, ca. 4-7 pill wide just above asci, long lapering with long necks papillate and slightl y raised above above. Asci cy lindrical to clavate sometimes venti'icose, stronia surface, appearing as dark bumps. Perithecial with spore-bearing part 22-29 X 4-5 pin, with stipe 13- walls composed of pale brown, thin walled, flattened 20 pm long. Ascospores hvaline, globose, to slightly 1)seudopai'enchvma, 16-20 pm thick. Paraphyses depressed globose, smooth, 2.8-3.1 pm diam. Ana- abundant, ca. 5.0-7.5 pill wide just above asci, long morph acroclontium-like occurring near base of stroma. tapering above. Asci with spore-bearing part 22.5- Colony grayish brown. Conidiophores verticillately 26.5(-29.5) X 3.5-4.5 pm, with stipe ca. 17-22 prri branched with primary and secondary whorls, conidio- long. Ascosporcs hyaline, globose to slightly dc- genous cells wide near base and tapering to fii'st coniclia, pressed globose, smooth, 3.0-3.7 pill diani. from first conidia to tip zigzag or spiral-like, denticulate. A namorpli. Not fhuncl on stroina. Conidia hyaline, ohovoid, 2.9-3.0 X 2.3-2.5 pm. ilalyitat. On dead wood. Habitat. On dead wood. Known di.ilrilyution.. Costa Rica, known onl y from Known distribution. Costa Rica, known only from the type collection. the type collection. Specimens examined: COSTA RICA. San Jose, San Speo incus examined: COSTA RICA. Puntai'cnas, Reserva Gerardo de t)ota, Albergue de Montana, Savegre, 5 kin Biologica Bosque Nuhoso Moii teverde, (an ru Cientifico SW Cerro de la Muerte; 9 33'2"N, 8348'27"W, elev. 2350 m, Tropical, Sendero El Camino, elev. 1536 In, t 0.3058. on wood, in forest of Quercus wpvensis and Q.ceemana, 19- —84,7933j, on dead wood, 5-XI-2003, SM. Huh ndur[ FA, X-1994. R.E. haling 7389, mi/h M. Ma/a, (Jmaña, Mueller Fernandez, SM1I4958 (hololype USJ. isotype F). cr' .5/jar (holotype USJ; isotvpe NY). Amplistroma tartareum I Tiuhnclorf & Samuels, sp. nov. Amplistroma ravum Ilitbndo ri,A.N. Mill., M. (;reif & MycoBank NIB 513245 Fic;s. 53-61 Samuels, sp. nov. FIGS. 41-52 Stroniata pulvinata cl tui'binata coiiilectivo lab. 10- MycoBank MB 513244 15 mm longa X 2-5 minlata )< 0.5-1 mm c'l'assa, cum Stroniata irregulariter pulvinata ConnectivO tab, 3-5 mm subiculo ad basim; Longii'osti'is; extus fulva vel umhi'ina et longa X 2-4 nun lata X 1-2 mm crassa, cum suhictilo art intus alha vet cremeas; lexitira tartareus. Pei'ithecia sub- hasini; tongirostris; extus fulvus vel ravus et intus lumens. glohosa 350-450 pm diani, 450-550 pill alto, inonosticha, Perithecia sithglobosa 350-450 pill diam. 450-550 pill alto, collis longis praeclita; ostiola papiltuta. Paraphvses abun- monosticha, collis loiigis piaedita; ostiola papillata. Paraph- clans, ca. 1-7 pin supra ascos, vel'Sus apic'eln perangusiani. vses ahundans, ca. 4-7 pm supra ascos, versus apiceni Asci cvlindrici vel clavati, pal's sporifer 22-29 X 4-5 piii. peranguslarn. Asci cylmdi'ici vel clavati, pars spoi'ifer 22-29 stipitati. 13-20 pill longi, octospori, uniseriat.i. Ascosporae X 4-5 pm, longi-stipitati, 13-20 pill longi, octospori, ilni- glohosae, 2.8-3.1 pin diam, hyalinae. seiiati. Ascosporac globosae. 2.8-3.1 pm cliam, hvalinae. EI'molo. tar/arms refers to crumbling surface LI'molo. ravu,s = yellowish gray, refers to the texture of the stroma. color of stroma. Subiculum restricted to immediate vicinit y of Subiculuin restricted to the immediate vicinity of stroma, yellowish gra\ to brown. Stroma widely stroma, yellowish gray to brown. Stroma turbinate to pulvinate to broadly turbinate on wide connective irregularly pulvinate on wide connective (ca. three- (three-quarters or greater width of stroma) , variable (Juarters width of strolna) , 3-5 mm long >< 2-4 ITIH1 in size but ca. 10-15 mm long X 2-5 111111 hi'oacl X broad X 1-2 111111 thick, perithecial mounds distinct; 0.5-1 111111 thick, ascomata mounds distinct; texture texture firm. With reflected light, stroma surface crumbly' when dry. With reflected light sti'oma surface yellowish, dark gray, flecked with white, rostrale with tail to yellowish brown, appearing white to cream with dark brown, long ostiolar necks curved or straight, disintegration of' outer wall, i'ostrate with dark brown, easily broken; interior dark gray. Under transmitted long ostiolar necks curved or straight, easil y broken; light stroma composed of brown, globose to angular interior cream. Under transmitted light stroma pseucloparenchyma; a hyphal layer of loosely woven, composed of loosely woven, h yaline to pale brown yellowish, textura intricata present above perithecia hvphae of textibra intricata; somewhat more densely between perithecial wall and upper strotnal surface; woven and slightly darker at the stroma surface, stromal surhuce hyphae composed of dark brown, contiguous with tlie iteck surface or necks smooth. densely woven tcxtul'a intl'icata: neck surface smooth. Perithecia sUl)globose, 250-300 pin cliam, immersed Pei'ithecia suhglobose, 450-550 pill high, 350-450 pm in su'oma or ill separate tlioundls, moriostichouis, with wide, immersed in stroma or in separate mounds, long necks raised above stronia surface, ca. 225-

IflI1\DORF II \I,: A l I/TIVIT'01 1, 1 (;1\. NOV.

4 c . W '.* •' '.J Me L1/ : .41 _____ 42 I11 . - 1-

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-: 60 •p• • ___ / at - - - •a J'.• •, - ,• •-'- - - -. (i.' '• t '.• -- _;•- • , . . 4!:,_., .- •(_ ç -'-:; h _58 61

Fi s. 41-52. :1 mp1iIiinta ravu n. 53-61. .4 in/i/is/mum /a,-Ia,iu in. 11 - 1 3, 53.S11- mata at I lu si thsi al c . 44, 56, 57. Asti. 45. 58. Ascospores. 1. Pataplisses. 47. (3aiidia. 48. Cotudiophores with couidia. 49, 51, 52. 59. Longitudinal SCCIIOI1 through stromal tissue. 50. 60. 1 .oiigiitidinal section through stroma .54,55. Stromatal surface. 61. Longitudinal section through ascomatal wall. 41-13. 53-55 by phototniciographv; 45, 49-52, 56-58 by DIC; 44. 46-48, 59-61 by I'I-l. Bar: 53 = 5 mm; 11-43, 54 = 1 55 = 500 tm; 50. 60 = 100 Mm: 44-49, 51 52, 56-59, 61 = if pm. 41-52 from SMH4958, 53-61 from Nur 5128. ,( (I (((.1

250 iiii luiig. 60-70 pm wulc. Pciitliccitl \vtlls cmii- II (IlllntI(/clla (ni(LTcLa1a (\\ alli. ) Sace., Svll. Futip. l)osccl of brown, thin walled, flattened pseudoparen- 1:455. 1882. Fi;s. 74-95 (hvma, 14-20 pm thick. Paraphyses remnants not well = .Sphaeria con gregata Walir., Flora Crypt. Germ. Sect. 2, preserved, presumed wide and tapering. Asci cylin- 4:786. 1833. drical to clavate, sometimes ventricose, with spore- Geratostoma sphaerospennurn Fuckel, Svmh. Myc. p 127. bearing part 17-25 X 2.5-3.5 pm, with stipe Ca. 9- 1869. 12 pm long. Ascospores hyaline, globose to slightly Geratostomehla splzaerospenna (Fuckel) Sacc., Syll. (4-pressed globose, smooth, 2.5-3.2 pm diam. Fung. 1:412. 1882. I namorph. Not found on substrate. Linostomeila sphaerosperma (Fuckel) Petrak, Ann. habitat. On decaying bark. Mycol. 23:42. 1925. known distribution. Singapore, known only from Ascomata globose to lageniform, short to long- type collection. beaked, not collapsing when dry, dark brown, surface Specimens examined: SINGAPORE. Botanic Gardens, on smooth, 325-375 pm diam, neck up to 425 pm long, Vep/zehium hark, 4-XIl-1919, M.Nur 5128 (holotype BPI superficial, occurring in large clusters or separate, 78926, isotype F). sometimes on sparse to abundant subicular hyphae. Subiculuin when present, occurring as extensive, Amplistroma xylarioides (Pat.) Huhndorf & Samuels, white, tomentose hyphae surrounding ascomata. (OtTh. nov. FIGS. 62-73 Ascoma wall opaque in surface view; in longitudinal Hypocrea xylarioides Pat, in Patonillard & Lagerheim, section 40-70 pm thick, composed of an inner layer Bull. Herb, Boissier 3:71. 1895. Basionym. of small, flattened, brown cells, a middle layer of small \IvcoBank MB 513246 to large, polygonal to irregular, hyaline cells that Subiculum not seen. Stroma irregularly turbinate collapse or rupture creating small empty pockets, and to obovoid on moderately wide connective (ca. one- an outer layer of small, brown cells. Asconiatal apex half width of stroma), about 6-7 mm long X 8- short to elongate beak-like, 250-425 pm high, 75- 9 mm broad X 6-7 mm thick based on dried stroma; 100 pm wide, composed of cells similar to ascoma wall texture firm, cartilaginous. With reflected light, cells, ostiole circular, with periphyses. Paraphyses stroma surface black with white to cream crustose abundant, Ca. 4-7 pm wide just above asci, long exudate, roughened papillate with dark brown, short tapering above. Asci thin-walled, cylindrical to clavate, ostiolar necks; interior dark brown. Under transmitted light, interior stroma composed of loosely woven sometimes ventricose, spore-bearing part 21-26 X 3- brown hyphae, tissue under stroma surface differen- 4.5 pm, with stipe 12-19 pm long, with eight ascospores uniseriately arranged. Ascus apical ring minute tiated into four layers by color and density, of closely woven, brown hyphae of textura intricata; hyphae to inconspicuous, not staining in Meizer's iodine somewhat more densely woven, slightly darker and reagent. Ascospores hvaline, globose to slightly not gelatinized at stroma surface (SS), thin outer depressed globose, appearing thick-walled, smooth, middle layer (OM) of hyaline hyphae, thick inner 2.3-2.8 pm diam. Anarnorph acrodontium-like. In middle layer (IM) of dark brown loosely woven hyphae culture colony growth 19-21 mm on CM and 21- and thick inner layer (IN) of hyaline to pale brown 23 mm on MEA after 7 d. Rapidly forming secondary loosely woven hyphae. Perithecia subglobose, 200- colonies. On CMA surface light brown (huff 45—rosy 425 pm diam, immersed, polvstichous in 3-4 layersjust huff 61), appearing powdery from abundant aerial, under stroma surface, with long necks papillate and sporulating conidiophores, surface hyphae sparse, slightly raised above stroma surface, appearing as dark reverse light brown (buff 45—rosy buff 61), no exudates, bumps. Perithecial walls composed of pale brown, thin submerged hyphae sparse. On MEA surface light brown walled, flattened pseudoparenchyma, 15-20 pm thick. (buff 45—light hazel 88 over time), reverse tan to brown Paraphyses not seen but presumed to be wide and long to olive over time (honey 64—isabelline 65—olivaceous tapering. Asci with spore-bearing part Ca. 19-20 X 3- 48) in center of colony, no exudates, hyphal appear- 4 pm, with stipe ca. 15-16 pm long. Ascosporeshvaline, ance same as on CMA. Conidiophores arising ortho- globose to slightly depressed globose, smooth, 2.5- tropically from light brown surface hyphae, erect, 2.9 pm diam. differentiated, light brown, with whorls of secondary Ananiorph. Not found on the stromata. and tertiary branching toward the apex, about 250- Habitat. On dead wood. 300 pm long, 3.5-5.0 pm wide at base. Conidiogenous Known distribution. Ecuador, known only from the cells arising in verticils or whorls on the branches, type collection. narrow lageniform, 7.0-10.3 X 1.8-2.2 pm, apex .Secimens examined: ECUADOR. Stir vieux troncs, San forming elongate rachis, proliferating sympodially, Jorge, Juillet 1892, leg. M. Lagerheim (holotype FH; isotype straight, flexuous, 6.0-27 X 0.7-1.0 pm, with denticu- N. late, conidiogenous pegs. Conidia light brown, oh-

Hut INDORF ET Al: A11111LISTROj1A GEN. NOV. 915

f4 Is

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- 4. jf. 1 ' ..c 'r

SSOM IM IN 64 65

p68 • 4? 67 _69 D i1J'1t. -

l-' J - IV

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70 71 72 .' U

Fius. 62-73. Arnpiictroma xylarioides. 62, 63. Strotiiata. 6-I. Longitudinal section through stroitial tissue showing four distinct layers (from left to right, SS = stromal surface, OM = outer middle, IM = inner middle, IN = inner). 65. Longitudinal section through ascoiriatal wall. 66. Longitudinal section through stroma. 67, 70. Paraphyses. 68, 69, 72. Asci. 71. Ascospores. 73. Longitudinal section through ascomatal neck. 62, 63 by photomicrography; 64, 66, 73 by BF; 65, 68, 69, 71, 72 by DIC; 67, 70 by PH. Bar: 62, 63 1 mm; 66 = 500 pm; 64, 65, 73 = 100 pm; 67-72 = 10 pm. All from NY isotype.

ovoid, 2.0-2.3 X 1.5-2.0 tim. For additional teleo- North Carolina, Macon County, Highlands, Blue Valley, morph description see Réhlová and Seifert (2004). 35 01'09"N, 8316'25"W, elev. 1000 in, on 5 cm branch, 7-X- Habitat. On dead bark and decorticated wood. 1996, S.M. Hu/indorf F.A. Ferndndez, SMH2 743 (F); Puerto Known distribution. Europe, United States (North Rico, Luquillo Mountains, El Verde Research Station, 16- Carolina, Puerto Rico, Tennessee). hectare grid, 18 19'37"N, 65 49'02"W, elev. 350-425 iii, on Specimens examined: GERMANY. Type: F. rh. 2013. Ad bark of 25 cm log of Citionanthus (loiningoeosir, 5-X-1995, lignum Pini svlv. putridum, rarissimo, Vere ca. Johannisberg S.M. Huhndorf SMHI760 (F), culture (CBS 124656); (holotype of C. sphaerospennum, G). UNITED STATES. Tennessee, Sevier County, Great Smoky Mountains National ,( )I.( )( L\

; -

I 77 I -" - -

74'-.; .fr j9i. 75 76 -

79 *i, 80 - 81

1'• s_ / iç • A .83 _84 _85 _86 3t Wit 88 89

90 I

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WL 93 94 95

Fi. i 1-9a. 11 (//I)UI/I/(Ila C()!l41etaIa. 71, 79-81, 87 Ascoin.tta on the substrate. 75, 82, 90. Longitudinal scctli)Ii Iluotigh asconia. 76, 86, 95. Longitudinal section through ascomatal wall. 77, 83, 85, 88, 89. Ascus. 78. Ascospores. 84, 94. Paraplivses. 91- 93. Con idiophores with coindia. 74, 79-81, 87 by photomicrograph y; 75-78, 82, 85, 86, 88, 90, 93, 95 by DIG; 83, 84, 89, 94 b y PH; 91.92 by SEM. Bars: 79, 80,87 = 1 mm; 74, 81 = 500 urn; 75, 82, 90 = 100 tIm; 76-78, 83-86, 88, 89, 93-95 = 10 lim; 91,92 = 5 lm. 74-78 from type of Ceratostroina Sph(ieroc/)('flnum, 79 from ANM8I, 80-86 1 10ff! SMH2743, 87-95 from SMH 1760.

HUHNDORF ET AL: AMpII,STRO'IA GEN. NOV. 917

Park, vicinity of Gatlinbiirg, Indian Gap, 35 36'34.58"N, ascospores are uniseriately arranged in a cylindrical 83 26' 19.2"W, elev. 1591 m, on wood fragment, 13-V1I-2004, spore-bearing part. In broken asci the shape of the A.N. Mi/fr,; SM. Hu/indorf, G.K. Mugarnbi, L. Ruiz- spore-hearing part can change and become ventricose Sanchez, A\A181 (F. ILLS). and the spores rearrange irregularl y in a biseriate htshion. Ascus stipes are difficult to measure because KEYTOANIPLISTRONIAI\( 1F they break easily and error in ascus size probably

Ascoinata formed in it stroma ...... 2 favors the short range for all species. Ascomata not stromatic, at times with a hyphal The overall closest morphological fit of the subiculum ...... Wallrot/i ic/ia con gregala anamorphs of A. carolinianum, A. ravurn and W. 2. Strorna texture cartilaginous when dry ...... S congregata is Acrodontium de Hoog. Acrodontinin is 2. Stroma texture soft or crumh1' when one of a number of anamorphic taxa that have dry ...... A mplislroina tartaren ni conidiogenous cells that bear conidia on a sympo- 3. Stronta with polvstichous ascornata ...... 4 dially proliferating rachis. Acrodoninim was separated 3. Stroina with monosticlious ascontata ...... from Beauveria Vuill. and iritirachium Limber to ...... Aniplistimnna ravUm accommodate taxa with rachides that proliferate 4. Stroma surface more or less smooth with straight and are denticulate (de Hoog 1972). ascomatal necks barely erurn pent...... Beauveria. and Trilirachin in bear conidia in it genic- ...... Amplistroma ha1/ingi 4. Stroma surface rough, mostly with ascomatal tilate or zigzag fashion on a proliferating rachis and necks distinctly erumpent ...... S they differ from each other in the presence of 3. Stromatal surface with white, cream to brown denticles in the former and cicatrizeci in the latter. A amorphous material, extensive or not ...... 6 further difference in Acrodontiurn species is the pres- 3. Stromatal surface without extracellular coating or ence of conidia with distinctly apiculate bases, although amorphous material ...... 7 de Hoog (1972) does describe occasional apiculate 6. Stronia surlace with extensive white to cream conidia occurring in some species of Beauveiia and amorphous covering, stromal hyphae distinctly iritirachium. Other characteristics that distinguish the brown and multilayered; ascospores 2.5-2.9 tut species are the branching of the conidiophores, shape of (ham ...... A inplistroma X'Vlarioides conidiogcnous cells and the colony color in culture. 6. Stronial surface with some brown amorphous Various combinations of these characteristics occur with covering, stromal hyphae pale brown and not iriijrachujn species tending to have lightly pigmented distinctly layered; ascospores 1.7-2.2 pni diam colonies and conidiophores with verticillately branch- ...... Ainplictroma diin,nutisporo ifl ing, narrow width conidiogenous cells and Beauveria Stronta dark brown, appearing black when dry, ii species tending to have whitish colonies and single interior dark ...... A mpiz.strorna gvianense 7 Stroma tan to dull brown, necks darker brown, branched or unbranched con idiophores and anipulli- interior cream to tan .... . Amplistroma (am/in/a fluin fbrm or flask-shape conidiogenous cells. Acrodoninim species tend to have lightly pigmented colonies; the DISCUSSION conidiophores are variously branched and the conidjo- genous cells may be narrow or flask-shape. Family Amplistromataceae resolves as a monophyletic The anamorph of A. carolinianuin resembles dade with strong bootstrap and Bayesian support with Acrodontiurn virell,i in (Fr.) de Hoog in its verticillate, LSU data, but its sister taxon relationship with the lageniform to ampulhiform conidiogenotts cells and Chaetosphaeriales is not supported. Two genera are apictilate, pale brown coniclia. It differs in faster recognized in the family based on molecular and growth, paler colony color and elongating conidio- morphological data and three of the species in phores (FIG. 14). The anamorph of W.suliicuiosa Amplistroma are placed in the genus based on resembles Acrodontium griseum (Fassat.) dc Hoog in its morphological data alone (A. diminutisporurn, A. verticillate, lageniforns conidiogenotts cells with long tart areurn, A. xylarioide.$). rachides and apicitlate, pale brown conidia. It differs in The two genera included in the family share similar its faster growth and tan to brown colony color; A. morphological characteristics. Walirothiella congregata griseum colonies are described as being green when and species of A rnplistroma share cyhindro-clavate, older (de Hoog 1972). In A. ravurn the anamorph is stipitate asci, minute, globose ascospores and wide known only from its presence on the base of the paraphyses that are long-tapering al)OVe the asci. teleomorph stroma. The conidiophores appear to be Except in the case of A. diminutisborurn, asci and irregularly branched and are not strongly differentiated ascospore sizes are not useful in distinguishing these from the vegetative hyphae. The long, narrow con idio- species. In all species the asci are long-stipitate and genoims cells (FIG. 48) are similar to those illustrated for when intact and attached to the hymcnium the Acrodonliurn hydni cola (Peck) de Hoog (de Hoog 1972).

9 1 ;\ 1 \( .( )t .( )( ; I.

The anamoiphic genera have been associated with is roughened with densely clustered ascornata necks, teleomorphs in different taxonomic groups. Many whereas in A. /ialhingii the surface is smoother and the species of JJeauveria occur on insect substrates and necks more widely spaced. In A. xylariou/es the surface is some have been connected with Cord'eps (Fr.) Link densely covered with a white, amorphous coating not teleomorphs (Clavicipitaceac) (Shimazu et at 1988, seen in the stromata sections. A inpiislroma (himinulis- Zengzhi ci at 2001, I luang et a] 2002). 'iritirahium poiim also appears to have some kind of brownish species are the reported anarnorphs for Trichosphaerella amorphous material on the surface between the a'ratophora (Hñhn.) E. Mull. (Niessliaceae) and Cep/ut- erumpent necks but differs from A. xylarzoides by lotheen savori C. Booth (Cephalothecaceae) both lacking the distinctive hyphal structure of the stroma. described from decaying wood. In both these cases In A. caroiiivanu.m the pale brown stromata surface is the descriptions and the illustrations show denticulate dotted with numerous, erumpent, darker brown necks con idiophorcs that are straight, not geniculate. Both of anti in A. guianensethe strornata are dark, with both the these characteristics would place the anamorph outside surface and interior appearing black. Tritirachium and in Acrodontiunt. From these reports A mphistromna ravu m also shares the firm-textured and the results obtained in this study Aerodonlium stromata wall seen in the previous species, but anainorphs could be considered to be widel y spread ascomala are found in a monostichous arrangement. throughout the ascom ycetes. However caution proba- Another difference is seen in the elongate, erumpeilt bly should be used in interpreting the report of the ascornata necks that are darker than the surrounding anamorph of C. savorvi. Booth (1961) stated that stroma surface. The separate ascomata mounds often ascospores never germinated and the colony obtained are clearly distinguishable, giving the appearance of was from "mycelium growing out from small pieces of clusters of ascomata united on a basal stroma. the ascocarp wall." He says that the anamorph "covered Stroinata tissue is cellular in time basal portions and the surface" of the wood substrate that was incubated in of loosely woven hyphae below the stronlata surface a moist chamber several months and that the ascolnata and surroundling the ascornata. The last species in the developed among this mycelium. It is possible that the genus, A. larlareuni, differs from the others in its incidental presence of coniclia on the ascomata wall loose, crumbly textured, more effuse stroma. The could have been the source of the anamorph mycelium stromatal tissue is of loosely woven hvphae that easily and that this might not be connected to the life-c ycle of fall apart when touched. It is more or less uniformly C. savrnyi. cream to brown throughout. Species of Amplistrorna and Waliroihiella differ iliroihiehia differs from Amphistrotna in its lack of mainly in the size, amount and texture of stroma stromnata tissues. Of the collections of I'V congregata, structures. Large, fleshy-gelatinous to cartilaginous only SMH2743 from North Carolina has white tornen- stromata with polystichous ascomata are found in A. tose hyphae surrounding the groups of ascomata on the carob nianuin, A. diminutisporu in, A. guianense, A. substrate. The rest of the collections formed ascomata hallingu and A. x'cla.rioides. Distinguishing among in small or large groups without obvious hyphal four of the species (A. guianense, A. h.ailingii, A. tomentnm. The ascomata are long-beaked as in A. dimin u.lisporurn and A. x iarioidec) can he problemat- rami in anti A. tarlareitmand the Puerto Rican collection all ic. All four species iniially- v were identified as A. obtained in culture (SMH 17 60) produced xylarioides because the differences in stromata mor- anamorph. phologies did not seem to warrant separation. When Some fratures of Amnphislroma and Waliroihiehba also sequence data showed A. guianense to be distinctly are shared with other groups in the Sordariomycetes. separate from A. ha/hugh, it seemed prudent, despite The large stromata with polystichous ascomata found in the lack of molecular data for A. diminutsporum and some of the Amphisironia species resemble those bnnicl A. xIarioides, to take into account the morphological in I'ach'1'trpeBerl. ex M.E. Barr, J.D. Rogers & Y.M.Ju differences and treat them all as separate species. The and Caniarops amorp/i.a (Boeclijn) Nannl. The two stromata hyphae in all these species are loosel y woven species of I'ac/ivirvpe were included in the LSU textura intricata. Amplisiroma xIarioides has the most phylogenetic analyses and were found to cluster within distinctive stroniata tissue with several layers corn- the Diaporthales near the Valsaceae and not close to posed of different arrangements of dark brown Am/distroina (FR;. 1). The genus had been classified in hyphae (FI(;. 64). In the remaining species the the Calosphaeriaceae (Fernandez et al 2004), but these stromata tissue is hyaline to pale brown with outer results (l() not support that. placement. Analyses have surface slightly darker in color and somewhat more found that C. amnorpha belongs in the Boliniales densely woven. These species differ from each other (Huhndorf et at 2005, Huhndorf and Miller 2008). in the morphology of the stromata surface. The surface The Aniplistromataceac represents a family that con- of A. (arolinian.um, A. diminulisporu in and A. guianense tains both taxa developing stromata and those with Pr

l-IUIINI)ORF ET AL: AMPIJS7I?Oi'L4 GEN. NOV. 919

nonstromatic ascomata. Both Pachytrvpe and Camars Bayesian inference of phylogenetic trees. 3.1.2. hnp:// are placed in groups that represent similar examples. inrbayes.csit.fsu.edu/download.php [Accessed 8 Aug Globose ascospores found in all species of Amplis- 20061. Ronquist F. 2001. MrBayes: Bayesian inference of troma and Walirothielia resemble those found in pimylogenetic trees. Biotnlorniatics 17:754-755. I-Iypwrea Fr. (Hypocreales), Horinosperrna Pcriz. & I luhndorf SM. 1991. A method of sectioning ascomycete Réblová (Chaetosphaeriales). Sacc. and Ascochalara herbarium specimens for light microscop y. Mvcologia In all these other taxa however the asci become 83:520-524. rnultispored and the globose spores are part-spores, Fernández FA. 1998. Neotropical Ascoirtycetes 7. formed by the disarticulation of eight ascospores. Caudafi,spora biapiculatis sp. nov. from Puerto Rico. The type specimen of Ceratostoma S/5haerOS1berIflurn Svdowia 50:200-204. and our recent collections match the collection of W. Greif M, Mugamnhi GK, Miller AN. 2008. Two new congregata illustrated by Réhlová and Seifert (2004). genera in the Magnaporthaceae, a new addition to Based on morphology, Réblová and Seifert (2004) (á'ratcsphaeria and two new species of Lentomitella. suggested that %'V congregata was not. congeneric with Mycologia 100:940-955. Wcuhiculoca Höhn., which was of uncertain placement Miller AN, Fernández FA. 2004. Molecular system- in the Sordariomycetidae based on LSU data. Sequence atics of the Sordariales: the order and the family I .asiosplmaeriaccae redefined. Mycologia 96:368-387. data from W congregala corroborates that they are not Lodge DJ. 2005. Neonopical closely related (Fi(;. 1). Garcia et al (2006) showed that Ascomvcetes 13. Cornz/.mlvina amid Lrythrornada, two W subiculosa is placed in the Coniochaetales and new genera from time Caribbean and elsewhere. Fungal classified the genus likewise. However the placement of Diversity 20:59-69. the genus in the Amplistromataceac follows the type 2008. A new species of Garnarops and species, W congregata and the generic classification of phvlogeneutic analysis of related taxa in the Bolinia- W sub-i culosa will have to he revised. ceae. N Am Fungi 3(7)231-239. Doi: 10.2509/ naf'2008.003.00715 http://pnmwfimngi.org ACKNOWLEDGMENTS Madclison WP, Maddison DR. 2006. Mesquite: a modular system for evolutionary analysis. Version '1.12. http:// This work was supported in part by NSF PEEl Grant mesqlliteproject.org (Partnerships for Enhancing Expertise in Taxonomy) DEB- Meyer RJ, Plaskowitz JS. 1989. Scanning electron microsco- 0118695. The authors are grateful for the aid of R.E. py of conidia antI conidial matrix of Trichoderma. HaIling, B.E. Overton, NY and C, who provided access to Mycologia 81:312-317. type and other specimens. and D.J. Lodge for fieldwork Posada D, Crandall K.\. 1998. Modeltest: testing the model assistance. Sequences were generated in the Pritzker of DNA substitution. Bioinfbrmatics 11:817-818. Laboratory for Molecular Systematics and Evolution at the Ranmhaut A. 2002. Se-Al: sequence alignment editor. http:// Field Museum of Natural History. tree.bio.ed.ac. uk/software/seal/ Ravner RW. 1970. A mycological color chart. Kew, Surrey: .ii'IRAll J RE C11E1) Commonwealth Mycological Institute. 34 p. Booth C. 1961. Studies of Pyrenornvcctes Vt. ilizelavia, with RéblovS M. 2006 Molecular systematics of (2eralosiomella notes on some allied genera. Mycol Papers 83:1-15. .sensu lalo and morphologically similar fungi. Mvcologia de Hoog CS. 1972. The genera Beauvera, Isarea, liz/ti- 98:68-9%. (1 chzunz and Acrodont,tzmn. Stud Mvcol 1:1-41 - Seifert KA. 2004. Crtptadeiphia (Trichosphacriales), Felsenstein J . 1985. Confidence limits on plivlogenies: a new genus for holomnorphs with Iiac/nsponurn an approach using the bootstrap. Evolution 39:783-791. anamorphs and clarification of tile taxonomic status Fernández FA, Lutzoni FM, Hulmiiclorf SM. 1999. Ide- of Wallrothielia. Mycologia 96:343-367. omoq)1i-ananmorph connections: the new pvrenomycc- Shimazn M. Mitsuhashi W, Hashimoto H. 1988. Cordyceps tous genus Carpoligna and its Pleurollieriurn. anamorph. brongniarti, sp. nov., the teleomorph of Beauveria Mycologia 91:251-262. lzrongniartii. Trans Mycol Soc japan 29:323-330. Rogers JD. Jti YM, Huhndorf SM, Umaña L. 2004. Swofford Dl.. 2002. lAUPz: phylogenetic analysis using Pararnphisphaeria costaricensis gen. et sp. nov. and parsimony ( :'and other methods). Version 4.0 blO. Pach'/trvpe rinwsa sp. nov. from Costa Rica. Mycologia Sunderland, Massachusetts: Smnauer Associates. 96:175-179. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins Garcia D, Stchigel AM, CanoJ, Calduch M, Hawksworth DL, DC. 1997. The Clustal X windows interface: flexible Guarro J. 2006. Molecular phylogeny of Con iochac- strategies for multiple sequence alignnienit aided by tales. Mycol Res 110:1271-1289. quality analysis tools. Nucleic Acids Res 25:4876-4882. Huang B, (Ion-Ru 1., Zhen-Gang L, Mei-7.heu F, Zeng-Zhi Zengzhi L, Chum-1.1 1., Huang B, Mcizheii F. 2001. Discovery L. 2002. Molecular identification of the teleomorph of and demonstration of the teleomorph of Beauveria Beauveria hassiana. Mycotaxon 81:229-236. bassiana (Bals.) Vuill., an important entomogenous 1-tuelseubeck JP, Mark PVD, Ronquist F. 2001. MrBayes: fungus. Chinese Sci Bull 46:751-754. p