Mycological Society of America

Harpellales in Newfoundland Aquatic Larvae Author(s): Robert W. Lichtwardt, Merlin M. White and Murray H. Colbo Source: Mycologia, Vol. 93, No. 4 (Jul. - Aug., 2001), pp. 764-773 Published by: Mycological Society of America Stable URL: http://www.jstor.org/stable/3761832 . Accessed: 20/06/2013 07:58

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

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

.

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

http://www.jstor.org

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions Mycologia,93(4), 2001, pp. 764-773. © 2001 by The MycologicalSociety of America,Lawrence, KS 66044-8897

Harpellales in Newfoundlandaquatic insectlarvae

RobertW. Lichtwardt1 disseminatingthe obligate fungal symbiontsto new Merlin M. White stream sites (Yeboah et al 1984, Moss and Descals Departmentof Ecology & EvolutionaryBiology, 1986, Labeyrieet al 1996). In the process of studying Universityof Kansas, Lawrence,Kansas 66045-2106 this unusual adaptation, we discovered and report here severalnew of Harpellales as well as new MurrayH. Colbo hosts and distributionsof previouslyknown species. Departmentof Biology, Memorial University, St. John's, Newfoundland,A1B 3X9 Canada MATERIALS AND METHODS

Abstract: Among nine Harpellales (Trichomycetes) The dipteraninsect hosts of Harpellales thatwe collected reportedfrom aquatic insectlarvae collected in New- included freshwaterlarvae of Simuliidae, Thaumaleidae and foundland,Canada, four new species are described: (nonbitingmidges), (solitarymidges), nymphsof the Plecoptera (stonefly) Leuctra.Most of Harpellomycesabruptus [living in Thaumalea verralli our specimenswere collected from 18-29 May 1999, and Thaumaleidae)], Orphella avalonensis [in (Diptera: subsequentlyfrom 13 Aprilto 30 August2000, in a number Leuctra ferruginea(Plecoptera: Leuctridae)], Stachy- of streamsites in the general vicinityof St. John's on the lina litoralis[in Telmatogetonjaponicus (Diptera: Chi- Avalon Peninsula of Newfoundland(TABLE I). Larvae were ronomidae)], and an unnamed species of taken fromstreams and other habitatswith the aid of nets (in Chironomidae) fromwhich an culturewas and strainers,or theywere picked directlyfrom substrates obtained. Other Harpellales were from Simuliidae such as leaves, sticks,and rocks.Specimens were placed in ( melusinae,Pennella simulii, Simuliomyces jars or plasticbags and kept cold untildissected in the lab- microsporus),Chironomidae (Stachylinarobusta), and oratory.Living fungifrom the dissected gut were studied an unnamed new genus in caddis flylarvae (Trichop- and photographed using phase-contrastoptics. Cultures were in of dilute tera). Low vagilityof Plecoptera and the occurrence attemptedby placing exposed fungi plates brain-heartinfusion agar or a tryptone-glucose-saltsagar of differentspecies of Orphellain the USA and Eu- medium,both containinga shallowdistilled water thatthe existedbefore the North overlayer rope suggest genus with added antibiotic solution Atlanticformed a barrieras the continents penicillin-streptomycin separated. (Lichtwardt1986). Selected fungalspecimens were also pre- It is hypothesizedthat vicariant speciation may have served in CTAB buffer(Gottlieb and Lichtwardt2001) for occurred in Orphellaas well as in Harpellomycesfrom subsequentDNA extractionand sequencing. Thaumaleidae larvae. Key Words: biogeography,Chironomidae, gut fungi,Leuctridae, Simuliidae, symbiosis,Thaumalei- DESCRIPTION OF NEW TAXA dae, Trichomycetes,Trichoptera Harpellomycesabruptus Lichtw., White et Colbo, sp. nov. FIGS. 1-8 Thalli 6-7 pLmdiametro, plerumque ad basem ramosi, INTRODUCTION series longas trichosporarumellipsoidalium, 20-33 X 7-11 p.m,sub emissione 2-5 appendiculis ornatas producentes. Newfoundland,Canada, like manyareas of theworld, Zygosporae ignotae. In proctodaeo larvarumThaumalei- is a rich source of aquatic insectsinfested with sym- darum (Dipterorum) affixi. biotic Harpellales (Trichomycetes),commonly called Thalli 6-7 pLmdiam, often branched at the base, gut fungi.It is now known that species of some har- producing long series of ellipsoidal trichospores 20- pellid genera in larvalblackflies (Simuliidae) mayoc- 33 X 7-11 pom that bear 2-5 appendages upon re- casionallygrow from the gut into the developingova- lease. Zygospores unknown. In hindgut of Thaumal- ries, resultingin adult blackfliesthat are sterilebut eidae (Diptera) larvae. whose ovaries are filledwith fungal cystswhich the Etymology.L. abruptus = steep, for the precipitous female "oviposits." This serves as a mechanism for cliffwhere the hosts were collected. Specimens examined. CANADA. NEWFOUNDLAND: Accepted for publicationJanuary 15, 2001. Seepage on cliffbordering Southside Rd., St.John's, New- 1 Correspondingauthor, E-mail: [email protected] foundland,47° 33.23' N, 52° 42.40' W. Microscopeslide NF-

764

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions LICHTWARDT ET AL: NEWFOUNDLAND HARPELLALES 765

TABLE I. Collection sites on the Avalon Peninsula, Newfoundland, with Harpellales

Site Stream Date Latitude/Longitude

NF-1 Beachy Cove Brook at Hughs Pond 18-V-99 47037.76' N, 52050.82' W NF-2 Beachy Cove Brook at WitchHazel Rd. 18-V-99 47035.82' N, 52050.84' W NF-3 Goat Cove Brook at TuckersHill Rd. 18-V-99 47036.08' N, 52°52.53' W NF-4 LittlePowers Pond Outlet 21-V-99 47033.99'N,52051.95' W NF-5 Millers Brook under Millers Rd. near Indian Meal Line 21-V-99 47°37.33' N, 52049.87' W NF-6 Clemens Pond Outlet on Bauline Line 21-V-99 47037.84' N, 52°49.48' W NF-7 Beachy Cove Brook at WitchHazel Rd. 21-V-99 47035.82' N, 52050.84' W NF-10 Beachy Cove Brook at Witch hazel Rd. 22-V-99 47035.82' N, 52050.84' W NF12 Middle Cove cliffat splash zone 23-V-99 47°39.11' N, 52041.83' W NF-12a Middle cove cliff,drip area away from splash zone 23-V-99 47°39.1' N, 52041.8' W NF-13 VoiseysBrook at VoiseysPark RecreationalArea 24-V-99 47°39.11' N, 52041.83' W NF-14 Millers Brook 24-V-99 47037.33' N, 52°49.87' W NF-19 Broad Cove Riverat outlet of Healeys Pond 27-V-99 47°34.41' N, 52°51.01' W NF-21 Beachy Cove Brook at Hughs Pond 27-V-99 47037.76' N, 52050.82' W NF-22 Seeping cliffat Southside Rd., St.John's 28-V-99 47033.23' N, 52042.40' W NF-23 Seeping cliffat Southside Rd., St.John's 29-V-99 47033.23' N, 52042.40' W NF-26 Goat Cove Brook 29-V-99 47°36.08'N, 52052.53' W9 NF-30 Beachy Cove Brook at WitchHazel Rd. 13-IV-00 47035.82' N, 52050.84' W NF-31 Beachy Cove Brook at Hughs Pond 17-IV-00 47037.76' N, 52°50.82' W NF-32 ForestField, Salmonier 1-V-00 47010' N, 53027' W NF-33 Beachy Cove Brook at WitchHazel Rd. 17-VII-00 47035.82' N, 52050.84' W NF-34 Beachy Cove Brook at WitchHazel Rd. 30-VIII-00 47035.82' N, 52050.84' W

23-5 (HOLOTYPE: FH) prepared froma Thaumalea ver- gospores attached obliquely and submediallyto the ralli Edwardslarva (Chironomidae,Thaumaleidae). Other zygosporophore. Attached to hindgut cuticle of specimens collected fromSites NF-22 and NF-23 (TABLEI) Thaumaleidae larvae (Insecta, Diptera). are fromsame host as the holotype. Harpellomyceshas also been found in Pennsylvania, The eccentricusLichtw. type species, Harpellomyces USA (unpubl), and in (Lichtwardtet al 1987). & was described from Thaumalea larvae Moss, spp. Commentson those species willbe found in the Dis- collected in northern Sweden and Wales (Lichtwardt cussion section. and Moss 1984). It differsfrom H. abruptus primarily by (1) the lack of branching, and (2) the unusual Orphella avalonensis White, Lichtw. et Colbo, sp. narrowness of parts of the thalli, in some cases mea- nov. FIGS. 9-15 suring only a few micrometers in diameter, with en- Thalli usque ad 900 ,umlongi, in axe principaliad basem ad largement at the apex where trichospores form. Ma- ramos multos curtossteriles producentes atque apicam ramos in fasciculoscellularum basalium ture thalli of H. abruptus are more uniform in di- aliquot producen- cellula basali aliquot cellulis genitalibusatque ameter, with narrowing found primarily in the basal tes, quaque terminalibuscirca 30-95 jim praedita.Trichosporae circa 6 branches. Trichospores of H. eccentricusmeasure 20- ,im diametro,cochleatae, e cellulis genitalibusexorientes 25 X 6-8 Jimand bear three broad appendages upon 20-22 X -6 Jim.Rarius corpora disseminationismaiora, release from the generative cell. Trichospores of H. cellulisgenitalibus 28 X 11 jim,cellulis terminalibus 80 jim have 2-5 and are 20-33 X 7- abruptus appendages longi, trichosporis11 Jimdiametro, cochleatis. Thalli in than those 11 jm; thus they are only slightly larger proctodaeo nympharumLeuctridarum (Plecopterorum) af- of the European species. Zygospores of H. eccentricus fixi;pars fertilisex ano procurrens. (which were not seen in H. abruptus) have been Thalli up to 900 Jimlong witha main axis bearing found in only one from Sweden. Two features multiple short sterilebranches at the base and api- and the of H. abruptus-the basal branching of thalli cally producing several branches that terminatein more variable number of trichospore appendages- clustersof basal cells, each producing severalgener- prompt us to emend the description of the genus. ative cells with terminalcells about 30-95 jim long. Harpellomyces Lichtw. et Moss emend. Lichtw., Trichospores about 6 jim wide, coiled, developing White et Colbo fromgenerative cells 20-22 X -6 j,m. More rarely Thalli unbranched or branched at the base, pro- disseminationunits much larger,with generative cells ducing distally a series of collarless trichospores with 28 X 11 jLm,terminal cells 80 Jm long, and tricho- 2-5 appendages, or conjugating and producing zy- 11 jim wide. Attached to hindgut of Leuctri-

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions 766 MYCOLOGIA

(

-iI 'I*"1% .

q.

r "P i; ... , A

t' i 'i :Il

*1 .8 1 ! j .1-3 *.1

FIGS. 1-8. Harpellomycesabruptus from a thaumaleidlarva. 1, 2. Mature trichospores.3, 4. Detached trichosporeswith 2 and 5 basal appendages, respectively.5, 6. Earlystages in developmentof trichosporesfrom generative cells. 7, 8. Branches produced at bases of thalli.Scale bar = 20 ,pm.

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions LICHTWARDT ET AL: NEWFOUNDLAND HARPELLALES 767

FIGS. 9-15. Orphella avalonensis from a stonefly nymph. 9-11. Sporulating heads of thalli and some detached dissemi- nation units with coiled trichospores. 12. Branches at base of main thallic axis. 13, 14. Dissemination units consisting of a generative cell (g) subtended by a small basal cell (b), an extended terminal cell (t), and a coiled trichospore (ct). 15. A rare large dissemination unit. Scale bars = 20 ,im.

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions 768 MYCOLOGIA dae (Plecoptera) nymphs, with fertile part projecting Stachylina litoralis Lichtw., White, and Colbo, sp. nov. from the anus. FIGS. 16-18 Etymology.From the Avalon Peninsula. Thalli 135-200 X 10-12 ,xm,in membranaperitrophica Specimens examined. CANADA. NEWFOUNDLAND: per basem subbulbosa affixi,trichosporas 4-8 per thallum Beachy Cove Brook at Hughs Pond, 18-V-99,47° 37.76' N, producentes,39-47 X 10-12 ,im, colare carentes.In mem- 52° 50.82' W. Microscopeslide NF-3-W7(HOLOTYPE: FH) brana peritrophicalarvarum Chironomidarum affixi. prepared froma Leuctraferruginea (Walker) nymph (Ple- Thalli 135-200 X 10-12 ,pm, attached to the peri- coptera,Leuctridae). Other specimenscollected fromSites trophic membrane by a slightlybulbous base; 4-8 tri- 2 and 3 (TABLE I) and fromsame host as the holotype. chospores produced per thallus, 39-47 X 10-12 pLm, Of the four other species of Orphella described, without a collar. Attached to peritrophic membrane only 0. helicosporaSantam. & Girbal (Santamaria and of Chironomidae larvae. Girbal 1998) produces tightlycoiled trichospores. Or- Etymology.L. litoralis = of the shore. phella haysii Lichtw. & M.C. Williams normally pro- Specimensexamined. CANADA. NEWFOUNDLAND: Mid- duces curved trichospores that are occasionally partly dle Cove cliffat high tide splash zone, N of St.John's, New- coiled. Orphella avalonensis differsfrom 0. helicospo- foundland,23-V-99, Site NF-12,47° 39.11' N, 52° 41.83' W. ra in several respects: the base of the thallus is not Microscope slide NF-12-1 (HOLOTYPE: FH) prepared froma larva Chi- bifurcate (FIG. 12), the diameter of the trichospore Telmatogetonjaponicus Tokunaga (Diptera, ronomidae). and terminal cell is greater, and the terminal cells Of the 30 currentlydescribed species of Stachylina, are cylindrical rather than clavate as in 0. helicospora. S. litoralis is most similar to S. macrospora Leger & The unusually large dissemination units that we Gauthier. The most obvious distinction is that tri- found (FIG. 15) may be anomalous. Two detached chospores of S. macrospora are narrower (7-8 ,Lm) anomalous dissemination units along with many oth- than those of S. litoralis.Another larva of T. japonicus er units of normal size were found with a thallus kept from the same population had a hindgut that was in a water mount for 3 d. One of us (MMW) has seen infested with a young, branched harpellid thallus, but similar dissemination units in an Orphella from Nova with no trichospores to allow for identification. Scotia, Canada. The host, T. japonicus, is one of relativelyfew Chi-

I I

FIGS. 16-18. Stachylinalitoralis from a marine chironomidlarva. Sporulatingthalli within the peritrophicmembrane. Scale bars = 20 pLm.

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions LICHTWARDT ET AL: NEWFOUNDLAND HARPELLALES 769 ronomidae frommarine habitats.Though located in the splash zone of the seashore, the larvae were bathed in freshwater flowingslowly down the cliff, suggestingthat while the host larvae toleratesea wa- ter,the gut fungusis not trulymarine. Another pop- ulation of T. japonicus larvae, collected on rocksthat are submergedat high tide at Maddox Cove south of St. John's,were not infestedwith any . Smittiumsp. FIGS. 19-21 In the hindgutof four Chironomidae larvae from Goat Cove Brook (Site NF-26) we found a species of Smittiumthat is probablynew, but we are not naming it at this time because of insufficientspecimens to fullydescribe it and clearly distinguishit from the other 55 species of the genus. The ellipsoidal tri- chospores [(14-)19-32 X 3-3.5 ,Im] witha median bulge and a collar 3-5 jim long do not fitdescrip- tions of other Smittiumspp. In the midgut of most of the chironomidlarvae we also found thalliof Sta- chylinarobusta Lichtw. & M.C. Williams (see below). An axenic culture (NF-26-3) of this Smittiumsp. produces abundant trichospores,a small number of which extrude their sporangiosporesfrom the spo- rangium wall (FIG. 21). An unusual feature is that sometimesthe extruded sporangiosporeproduces a germ tube thattwists and at timeswraps around itself (FIG. 21). There is no evidence that these extruded spores successfullyestablish new colonies in culture.

HOSTS AND DISTRIBUTION OF PREVIOUSLY KNOWN SPECIES

Harpella melusinaeLeger & Duboscq FIGS. 22, 23 Harpella melusinaegrows attached to the larvalper- itrophicmembrane of a wide range of blackflyspe- cies. It is one of the most widespreadspecies of Har- pellales, though it has not yet been reported from Neotropical regions nor fromsouthern South Amer- ica where two other species of Harpella, namely H. tica and H. meridianalis,are common (Lichtwardt and Arenas 1996, Lichtwardt1997, Lichtwardtet al 1999, 2000, White et al 2000). Its unbranched thalli produce two to eight or more curved to coiled tri- chospores (FIG. 22), and it is most easily distin- guished fromthe other three species of Harpella by its tapered holdfastclearly seen in FIG. 23. We found H. melusinaeat Sites NF-3,4, 5, 13, 19, 30, 31, 32, 33, FIGS. 19-21. Smittiumsp. fromthe hindgutof chirono- midlarvae. 19. Partof a branched thal- 34, where hosts included Prosimuliummixtum Syme densely sporulating lus. 20. Trichospores.21. Axenicculture NF-26-3 showing & Davies, Simuliumvenustum/verecundum complex, unusualdevelopment in culture;in middlean unextruded mutata Simulium vittatum Stegopterna (Malloch), trichospore,on righta sporangiosporeextruding from its complex, and possiblyother simuliid species whose , and on left a germinated sporangiospore identitywe did not record. Frostand Manier (1971) whose germ tube has wrapped around itself.Scale bars = reportedfinding H. melusinaein more than fourspe- 20 ,im.

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions 770 MYCOLOGIA

Our Newfoundlandmaterial fits the descriptionof P. simulii,allowing for a slightlybroader range of measurementsthan the original description. Trichosporeswere 28-38 X 7-10 ,um,zygospores 74- 89 X 16-19 pLm,and zygosporophores26-45 X 13- 19 pLm. Stachylinarobusta Lichtw. & M.C. Williams FIGS.31-33 i This species was recentlydescribed (Lichtwardt and Williams 1999) fromPsectrocladius sp. (Chiron- omidae) in the RockyMountains of Colorado. Our l Newfoundlandcollection came froman unidentified chironomidlarva fromSite NF-3. Trichosporesmea- sured 30-40 X 6-8 ,Lm,and the robust thallus ta- pered to a narrowholdfast (FIG. 32) as in the Colo- rado specimens.

OTHER COLLECTIONS

Among the identifiabletrichomycetes not reported above, we include the followingrecord: Simuliomyces FIGS. 22, 23. melusinae within a simuliid Harpella peri- microsporusLichtw. in a Prosimuliummixtum larva trophic membrane. 22. A thallus producing two tricho- from Site NF-13. Various unidentifiablespecies of spores. 23. Typical tapered holdfastattached to the peri- Smittiumwere found in chironomidsand simuliids trophicmembrane. Scale bars = 20 pLm. (Stegopternamutata, Prosimulium mixtum, Simulium vittatumcomplex), as were Paramoebidiumspp. in cies of simuliidlarvae frommany stream sites in New- various simuliidspecies fromseveral sites (including foundland. P. curvumLichtwardt in Simuliumvenustum/verecun- Pennella simulii M.C. Williams & Lichtw. FIGS. 24-30 dum complex from Site NF-34, and in another si- This common inhabitant of blackfly hindguts in muliid fromSite NF-14), and in Ephemerellaaurivillii Newfoundland was found at Sites NF-1, 3, 4, 5, 6, 10, Bengtssonfrom Site NF-3.An unidentifiedgenus was 13, 14229, 30, and 31. Hosts included Prosimulium also found in caddis flylarvae (Trichoptera)belong- to the a discoverednew mixtumand Stegopternamutata among other species ing Limnephiloidea, recently of hosts for not identified.Pennella simuliiwas originallyreport- Harpellales (White 1999). ed fromthe RockyMountains of Colorado (Williams and Lichtwardt1971) and is illustratedhere because DISCUSSION thereare fewadequate photomicrographsof the spe- cies. The trichospores(FIGS. 24, 25) and zygospores The relativelycool water temperaturesof streams (FIGS. 26-28) of Pennella are verysimilar to the rel- duringour major collectionsin May,1999 (10.5-18.5 ativelycommon and more widespread species Genis- C, av. 15.4 C) provided a diversityof aquatic insect tellosporahomothallica, also a blackflygut inhabitant, larvae. In turn, this led to our findingmore than withthe primarydistinction being the nature of the eight species of Harpellales, four of them new, and holdfast.Genistellospora spp. have a well-defined,sol- several species of Paramoebidium(Amoebidiales). id holdfastwhereas Pennella spp. have simple to bi- Members of the Amoebidiales,formerly considered furcatebasal cells attached to the hindgutcuticle by to be Trichomycetes,have now been shown convinc- means of a mucilaginous secretion from the basal inglyto be protozoansrather than fungi(Benny and cells (FIGS. 29, 30). O'Donnell 2000). Frost and Manier (1971) reported findingP. hov- Only two previouspapers reportedTrichomycetes assi Manier in Newfoundland blackflies,a species in Newfoundland.Frost and Manier (1971) found previously known only in . We cannot discount Harpella melusinaeand a Pennellathat they identified the presence of P. hovassiin Newfoundland,but the as P hovassi.Yeboah et al (1984) investigatedseveral identificationappears to have been made by Manier kindsof fungalcysts produced by ""in from preserved material,which may have distorted the ovaries of adult blackfliesin the area where we some of the measurements. conducted the studiesreported here. Moss and Des-

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions LICHTWARDT ET AL: NEWFOUNDLAND HARPELLALES 771

I 1

io

*A v .!ifli2i_v '.i FIGS.24-30. Pennella simuliifrom simuliid hindguts. 24, 25. Trichospores,the released ones withbarely visible multiple appendages. 26-28. Zygospores.29, 30. Two variantsof holdfastcells thathave secreteda mucilaginoussubstance. Scale bar = 20 pLm.

cals (1986) in Great Britain showed conclusively that gions. These stoneflies are among the least vagile of such cysts were a stage of Harpellales development. aquatic . Three Orphellas in southern Europe In their study the cysts belonged to Harpella melusi- include the type species, 0. coronata Leger & Gauthier nae. Our own investigations on harpellid ovarian cysts in France (in Nemouridae), and two species in in Newfoundland will be reported in another article. recently described by Santamaria and Girbal (1998), Of special interest is the distribution of species of 0. catalaunica and 0. helicospora (in Leuctridae). In Orphella and Harpellomyces.Orphella spp. are known North America, Williams and Lichtwardt (1987) to occur only in nymphs of three sisterfamilies of Ple- found Orphella haysiiLichtw. & M.C. Williams in Nem- coptera: Nemouridae, Leuctridae, and Capniidae. The ouridae from a Colorado Rocky Mountain stream. A families belong to the Suborder Arctoperlaria whose midwestern species, 0. hiemalis Peterson, Lichtw. & species are restricted to Nearctic and Palearctic re- Huss, was described from Capniidae (Lichtwardt et al

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions 772 MYCOLOGIA

FIGS.31-33. Stachylinarobusta thalli sporulating within the peritrophicmembrane of a chironomidlarva. Scale bars = 20 aLm.

1991, 1993). The new species, 0. avalonensisin Leuc- Thaumaleidae larvae another Harpellomyces sp. in tridaefrom Newfoundland, is the thirdNorth Ameri- two seepage sites in Pennsylvania, USA, that is similar can species. White (unpubl) has found what may be to the species in Newfoundland.None of the North the same Orphellaspecies in Nova Scotia, Canada. American collections has included zygosporesthat Thus, there exist six differentspecies of Orphella, would provide additional evidence of differencesor three from southern Europe and three fromNorth similaritybetween species on the two continents.To America. The low vagilityof the host stonefliessug- complicate distributionpatterns, Lichtwardtet al geststhat the genus Orphellaexisted in Arctoperlarian (1987) reported findinga species of Harpellomyces hostsprior to severanceof land connectionsacross the similar to H. eccentricusfrom Thaumalea sp. larvae at NorthAtlantic during the Lower Eocene (Brownand a waterfallin Japan. Lomolino 1998) and was followedby vicariantspeci- A better understandingof the evolution and dis- ation on the two continents. tributionof such gut fungiand theirinsect hosts will The thaumaleidhosts of Harpellomycesperhaps are depend upon a focused attemptto obtain specimens more vagile than stoneflies,but vicariance may also in theirparticular habitats from different geographic have contributed to speciation of their gut fungi. regions. ManyThaumaleidae are ecologicallyconstrained and often occur in seepage habitatsnear waterfallsand ACKNOWLEDGMENTS roadcuts (Sinclair 1996). The typespecimen of Har- We are grateful to National Science Foundation PEET pellomyces,H. eccentricus,was producing zygospores award DEB-9521811 for supportof thisresearch which was in addition to trichosporeswhen firstfound in north- also funded in part by an award fromthe DEEB to MMW ern Sweden in an unknownspecies of Thaumalealar- in honor of Dr. AJ. Mix. We thank Patricia M. Eckel for va. Subsequently, H. eccentricuswas discovered in lar- translatingthe new diagnosesinto Latin,and we appreciate KristinPeterson that vae of another Thaumalea sp. in Wales (Lichtwardt suggestionsfrom MatiasJ. Cafaro and the and Moss 1984), an allopatric distributionthat no improved manuscript. doubt is attributableto collecting bias. Thaumalea verralli, host of H. abruptus in Newfoundland, is a LITERATURE CITED species thathas successfullydispersed and colonized BennyGL, O'Donnell K. 2000. Amoebidiumparasiticum is a areas across the North Atlantic (Sinclair 1996). In protozoan, not a Trichomycete.Mycologia 92:1133- North America, Ferrington (unpubl) has found in 1137.

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions LICHTWARDTET AL: NEWFOUNDLANDHARPELLALES 773

Brown,JH, Lomolino MV. 1998. Biogeography.2nd ed. Harpellales from Sweden and Wales. Mycotaxon 20: Sunderland, Massachusetts:Sinauer Associates, Inc. 259-274. 691 p. , Peterson SW, Huss MJ. 1991. Orphellahiemalis: a FrostS, ManierJ-F.1971. Notes on Trichomycetes(Harpel- new and rare trichomyceteoccurring in winter-emerg- lales: and Genistellaceae) in larvalblack- ing stoneflies(Plecoptera, Capniidae). Mycologia 83: (Diptera: Simuliidae) fromNewfoundland. Can J 214-219. Zool 49:776-778. , WilliamsMC. 1999. Three Harpellales that live in GottliebAM, LichtwardtRW. 2001. Molecular studiesof se- one species of aquatic chironomidlarva. Mycologia91: lected Harpellales (Trichomycetes).Mycologia 93:65- 396-399. 80. Moss ST, Descals E. 1986. A previouslyundescribed stage in Labeyrie ES, Molloy DP, LichtwardtRW. 1996. An investi- the life cycle of Harpellales (Trichomycetes).Mycolo- gation of Harpellales (Trichomycetes)in New York gia 78:213-222. State blackflies(Diptera: Simuliidae).J InvertPath 68: SantamariaS, GirbalJ. 1998. Two new species of Orphella 293-298. fromSpain. MycolRes 102:174-178. Sinclair BJ. 1996. A reviewof the Thaumaleidae LichtwardtRW. 1986. The Trichomycetes,fungal associates (Diptera: of easternNorth America, a of .New York:Springer-Verlag. 343 p. ) including redefinitionof the genus Ent Scand 27: . 1997. Costa Rican gut fungi (Trichomycetes)in- Androprosopa. 361-376. fectinglotic insectlarvae. Rev Biol Trop 45:1349-1383. WhiteMM. 1999. Legerioides,a new genus of Harpellales in , Arenas J. 1996. Trichomycetesin aquatic insects isopods and other Trichomycetesfrom New , fromsouthern Chile. Mycologia88:844-857. USA. Mycologia91:1021-1030. , FerringtonLCJr, L6pez Lastra C. 1999. Trichomy- , Cafaro MJ,Lichtwardt RW. 2000. Arthropodgut cetes in Argentineanaquatic insect larvae. Mycologia fungifrom and summaryof tropicalTri- 91:1060-1082. chomycetesworldwide. Caribbean J Sci 36: (In press). Huss MJ,Williams MC. 1993. studies , Biogeographic Williams MC, LichtwardtRW. 1971. A new Pennella (Tri- on in winter trichomycetegut fungi stoneflynymphs chomycetes)from Simulium larvae. Mycologia63:910- of the genus Allocapnia.Mycologia 85:535-546. 914. , KobayasiY, Indoh H. 1987. TrichomycetesofJapan. , . 1987. Two new trichomycetespecies from Trans Mycol Soc Japan 28:359-412. Zapada spp. (stonefly)nymphs with an unusual distri- , L6pez Lastra C, Mazzucchelli MG. 2000. Fungi liv- bution. Mycologia79:473-478. ing in the gutsof larvalaquatic insectsin northwestern Yeboah DO, Undeen AH, Colbo MH. 1984. Phycomycetes Argentina.Mycologia 92:332-340. parasitizingthe ovariesof blackflies(Simuliidae). J In- , Moss ST. 1984. Harpellomyceseccentricus, an unusual vertPath 43:363-373.

This content downloaded from 134.153.186.189 on Thu, 20 Jun 2013 07:58:48 AM All use subject to JSTOR Terms and Conditions