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Structure and Development of Cleistoiodophanus Conglutinatus Gen

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Structure and Development of Cleistoiodophanus Conglutinatus Gen Amer. J. Bot. 63(6): 838-844. 1976. STRUCTURE AND DEVELOPMENT OF CLEISTOIODOPHANUS CONGLUTINATUS GEN. & SP. N. (ASCOBOLACEAE)l JOSE L. BEZERRA2 AND JAMES W. KIMBROUGH Departmentof Botany,University of Florida, Gainesville32611 A B S T R A C T Cleistoiodophanusrepresents a new coprophilousgenus of the tribe Iodophaneae in the Ascobolaceae (Pezizales). The only speciesthus far discovered,C. conglutinatus,is described and illustrated.Aspects of its cytologicaldevelopment are describedfrom cultures obtained fromapothecia found on sheep dung near Gainesville,Florida. Plasmogamyoccurs in aco- gonialcoils, two or threecells of whichgive riseto ascogenoushyphae. Ascogoniaare quickly enclosedby vegetativehyphae and the ascocarp continuesin a cleistohymenialdevelopment. UnlikeIodophanus and relatedgenera, the excipulumremains intact even afterspore matura- tion and the asci push throughthe epihymenialregions to release spores. The asci are charac- teristicallythickened at theirapices, diffuselyamyloid, and somewhatsaccate. The asci are predominantly8-spored, but have been foundwith four or 16 sporesper ascus. A previously undescribedOedocephalum imperfect stage was inducedin culture. IN A SEARCH for coprophilous discomycetesin manentia, albo colore praedita; excipula ectalis Gainesville,Florida, a funguswith lightcolored, cyanophilica,cellulas rectangularesvel quadratas roundishapothecia was foundto formabundantly habens; excipulamedularis cyanophilica, cum tex- on sheep dung when placed in a moistchamber. tura globulosa; asci generaliteroctospore, diffuse Cultures exhibitingan Oedocephalum imperfect amiloides,large clavati, cum parietibuscrassis in stage were easily establishedfrom fragmentsof juventute,pedicelati; ascospori uninucleati,hy- apotheciatransferred to agar media. Microscopic alini, ellipsoides,laeves, perisporiummucilagino- examinationof squash mounts and cryostatsec- sum ostentantes;paraphyses ramosae, hyalinae, tions of apothecia revealed a typicaldiscomycete septatae,multinucleatae. hymenium,but the asci remainedcovered by the Momentumconidiale: Oedocephalum Preuss. excipulumeven when fullydeveloped. The color Species typus: C. conglutinatussp. n. and externalmorphology of the apothecium,the Derivation: Greek, kleistos= closed, plus texturaglobulosa of the medullaryexcipulum, the lodophanus,a closelyrelated genus. natureof the asexual stage togetherwith the dif- Apothecia gregariousto cespitose, developing fuselyamyloid reaction of the asci, and the large, cleistohymenially,remaining closed, lightcolored; ellipsoidal and hyaline ascospores related the ectal excipulumof texturaprismatica, cyanophilic, fungusvery closely to lodophanus. However,the of rectangularor quadraticcells; medullaryexci- cleistohymenialascoma remaining permanently pulumcyanophilic, of texturaglobulosa; asci usu- closed, the saccate, stalkedand thick-walledasci, ally 8-spored,diffusely amyloid, broadly clavate, the smooth ascospores (present only in I. dif- thick-walledin youth,stalked; ascospores uninu- formis), and the differenttype of ascogonia cleate, hyaline,ellipsoid, smooth, a mucilaginous eliminatedlodophanus as a genusto accommodate perisporepresent; paraphyses branched, hyaline, the present species. Thus a new genus is pro- septate,multinucleate; conidial stage: An Oedo- posed for this unique Discomycete: cephalumPreuss. Coprophilous. Cleistoiodophanusgen. n. Cleistoiodophanusconglutinatus sp. n. (Fig. 1-32). Apothecia coprophila, gregaria vel cespitosa, sese cleistohimenialiterevolventia, occulsa, autem, Apothecia gregariavel cespitosa, sese cleisto- himenialiterevolventia, primo globosa, deinde 1Received for publication25 June 1975. irregulariterlobato-sulcata, occlusa, autem,man- 2Permanent address: Departmentof Mycology,Uni- entia,_colorecarneo vel albo-flavopraedita, ob- versidadeFederal de Pemambuco,Recife, PE, Brazil. scuriorapost maturitatem,leviter tomentosa, 0.5- The authorswish to acknowledgethe assistanceof Dr. ectalis, 20-35 tm crassa, F. B. Leal (UniversidadeFederal de Pemambuco,Bra- 3.0 mm diam; excipula zil) in supplyingthe Latin diagnosis,and that of Dr. cyanophilica,cellulas habens protensas, in 2-4 MeredithBlackwell and Dr. Gerald Bennyfor darkroom straturis,rectangulares vel quadratas, angulatas, assistance. x 6.0-10.5 jum,cum tex- Florida AgriculturalExperiment Station Journal Series multinucleatas,6.0-22.0 No. 5955. turaprismatica, haec cellulae suntlateraliter com- 838 July, 1976] BEZERRA AND KIMBROUGH-CLEISTOIODOPHANUS CONGLUTINATUS 839 pressae et externe hyphis hyalinis circumdatae, 21 x 11-16 jm; denticlesminute, sometimes in- quae apotheciis aspectum tomentosumofferunt; conspicuous,varying in numberfrom 5-16, 1 jum excipula medulariscyanophilica, cum texturaglo- long; conidia subglobose to ellipsoid, botryose, bulosa et cellulisellipsoidibus, 15.0-27.0 x 13.5- solitary,slightly verrucose, 8.5-13.5 (-15.0) x 15.0 tm;asci octospori,aliquando 4 vel 16 sporos 6.0-9.0,um,formed as blown-outends of the apex habentes,diffuse amiloides, large clavati,in apice of the denticles,germinating readily by 1-2 germs rotundi,cum parietibuscrassis in juventute,ped- tubes. icelati, (83.0-) 90.0-120.0 x 25.0-30.0 (-37.5) jum; ascospori uninucleati,distichi vel tristichi, Derivation: Latin,conglutinatus = unitedfirmly hyalini,large ellipsoides,in polis rotundi,laeves, together,referring to the arrangementof the ad maturitatemparietes tenues ostentates,15.0- apothecia. 19.0 x 9.0-13.5 tum,cum perisporiomucilaginoso; paraphyses hyalinae, ramosae, septatae, multi- Holotype: On sheep dung,Veterinary Science nucleatae,leviter in apice dilatatae,3-6 Km diam. Farm, Universityof Florida Campus, Gainesville, Momentumconidiale: Oedocephalum Preuss: Alachua County, Florida, Jan. 18, 1973, J. L. conidiophorisese ab hyphisrepentibus elevantes, Bezerra (FLAS-49579). tanquamrami laterales, erecti, simplices, cylindrici, septati, hyalini, 37.0-135.0 x 4.5-7.5 m; septi, MATERIALS AND METHODS-Culture and devel- 2-5, non constricti,primus in conidiophoribasi opment-Pellets of sheep dung were collectedon formatuset postremusin basi vesiculae; vesicula January18, 1973, at theVeterinary Science Farm, terminalis,aliquando intercalaris,pyriformis, den- Universityof Florida Campus, Gainesville,Flor- ticulata,17-21 x 11-16 jm; denticuliminuti, ali- ida. The materialwas placed in a moistchamber quando inconspicui,5-16, 1 Kmlong; conidia sub- at room temperatureand apothecia of the fungus globosa vel ellipsoidia,botryosa, singularia, leviter were detectedsix days afterincubation (Fig. 16). verrucosa, 8.5-13.5 (-15.0) x 6.0-9.0 m, ut Squash mountsand cryostatsections were made apicum denticulorumdilatationes formata, 1-2 of the apotheciaand examineddirectly in wateror tubisgerminativis propere germinantia. stainedin cottonblue (Korf, 1973), 5 % aqueous In arietis faecibus. Typus: J. L. Bezerra, Congo red (Kimbrough,1969), and Melzer's Re- Gainesville, Florida, America septentrionalis agent (Korf, 1973). Nuclear stainingwas done (FLAS F-49579). by the HCl-Giemsa methodas describedby Fur- Apothecia gregariousto cespitose, developing tado (1968) and by the aniline-bluemethod of cleistohymenially,globose at first,becoming ir- Tu & Kimbrough(1973). Cultureswere estab- regularlylobulate-sulcate but remainingclosed, lished by washing some apothecia in steriledis- flesh-coloredto light yellow, darker on drying, tilled water and placing apothecial fragmentson finelytomentose, 0.5-3.0 mm in diam; ectal exci- agar media. Aftercompletion of the studies,the pulum 20-35 Kmthick, cyanophilic, of 2-4 layers dung pelletsbearing apothecia were driedat 60 C of elongate, rectangularor quadratic, angular, and accessionedto the Universityof Florida My- multinucleatecells, 6.0-22.0 x 6.0-10.5 pm, a cologicalHerbarium. texturaprismatica, laterally compressed and ex- ternallycovered by hyalinehyphae giving a tomen- OBSERVATIONS-Cultural characteristics-The tose appearance to the apothecia; medullaryex- fungusdevelops slowly and formsscanty myce- cipulum cyanophilic, of textura globulosa, of lium on mycologicalagar (MA), potato-dextrose- globose to ellipsoid cells, 15.0-27.0 x 13.5-15.0 agar (PDA), cornmeal-malt-yeastextract-agar jum;asci 8-spored,sometimes 4 or 16-spored,dif- (CMMY), V-8 agar, V-8 agar plus dung extract, fusedlyamyloid, broadly clavate, rounded above, and dung extract-oatmeal-agar(DOA). Conidia thick-walledin youth, stalked below, (83.0-) are formedabundantly on all testingmedia but 90.0-120.0 x 25.0-30.0 (-37.5) ym; ascospores only sexual initials,and immatureapothecia are uninucleate,bi- or tri-seriate,hyaline, broadly el- produced. lipsoid,rounded at the tips,smooth, thin-walled at maturity,15.0-19.5 x 9.0-13.5 Am, a mucilagi- Mycelium-The vegetativehyphae are consis- nous perispore present; paraphyses hyaline, tently branched and multinucleate. Woronin branched,septate, multinucleate, slightly inflated bodies are observednear the septa. at thetip, 3-6 jumin diam. Conidial stage, an OedocephalumPreuss: co- Conidiophoresand conidia-The conidiophores nidiophoresarising as lateral branches fromre- arise as small protuberancesfrom the repenthy- pent hyphae, erect, simple, cylindrical,septate phae. After a period of elongation,an apical hyaline,37.0-135.0 x 4.5-7.5 m; septa in num- swelling develops at the tip of the developing ber of 2-5, non-constricted,the firstone formed conidiophoreswhich gives rise to the ampulla at thebase of theyoung conidiophore and the last (Fig. 3). Thornlikedenticles then form over the one at thebase of the ampulla; ampullaeterminal,
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