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C. , S. (2). . The a, IL 61801, 61801, a, IL S. juvenile has brown, species are placed 2012). In this study Cordana abramovii C. pauciseptata 1 Key words: Aquatic fungi Ascomycoyta Cancellidium Cordana Sporoschisma submerged woody debris S. saccardoi and S. parcicuneatum Cordana , uniseptata (15 collections) have previously been reported (9), IMA FUNGUS · 5(2): 425–438 (2014) 425–438 · 5(2): FUNGUS IMA (Cannon & Kirk 2007). Sporoschisma were collected from through al. 2006, Zhao et al. was found in 33 of 86 collections from was found in 33 of 86 collections from respectively, respectively, , and Carol A. Shearer , and Carol Cordana these are new 3 Cordana abramovii , (13), S. juvenile Another dematiaceous fungus, closely resembling Another dematiaceous fungus, closely Several species of 5-septate, doliiform, smooth walled conidia; one 28S sequence was generated from a Peruvian specimen one 28S sequence was generated from two 28S sequences and the identity was corroborated with Thai material. generated from Cordana abramovii, a range of sites. possibly due to the cushiony type is described as acervular, structures and setae appearance of the aggregated sporing The majority of the taxa on the substrate (Preuss 1851). as such; rather, belonging to the genus are not described conidia simply form on erect conidiophores with surrounding The Peruvian specimens of setae. questionable nature of the culture from which the DNA was which the DNA questionable nature of the culture from et extracted (Yeung Sporoschisma uniseptata has 1-septate, rarely 2-septate, reddish brown, verruculose conidia; musae and C. pauciseptata, from Perú (Matsushima 1993). saccardoi (PE0053) are characterized by pale brown to brown, cylindrical, septate conidiophores with swollen conidiogenous zones; terminal and intercalary polyblastic conidiogenous cells; and golden brown to dark brown, 1-septate, thick- additional species, Two walled, verruculose conidia. in the family Cordanaceae multiple sites, including S. , Andrew N. Miller , 1,2
), These taxa are reported herein from the These taxa are reported herein M. hemipsila and S. saccardoi, being made. has was . With the exception of S. saccardoi, . With the exception of , which was , Huzefa A. Raja , Huzefa 1 C. pinicola (PE0063) were S. saccardoi . However, they noted . However, has previously been linked to Melanochaeta hemipsila Sporoschisma hemipsila C. applanatum , Christine Do , Christine 1 Cancellidium applanatum and Sporoschisma saccardoi. Hypocreales S. saccardoi Sporoschisma saccardoi, S. juvenile, Hypocreales was dubious due to the S. uniseptatum, and ,
, Julia A. Balto , Julia 1 Cancellidium applanatum Cordana abramovii
in Kobe, Japan. A anelevationalgradientinPerútheDistricts of freshwaterascomycetesconductedalong survey (2007) divided the freshwater mitosporic fungi et al. (2007) divided the freshwater mitosporic et al. 2004, Pinnoi et al. 2006, Pinruan et al. 2007, et al. 2012). In this study in the Neotropics, multiple Cancellidium Department of Chemistry and Biochemistry, 457 Sullivan Science Building, University of North Carolina, Greensboro, NC 27402-6170, USA University of North Carolina, Greensboro, 457 Sullivan Science Building, and Biochemistry, Department of Chemistry 61820, USA Champaign, IL University of Illinois at Urbana-Champaign, Survey, Illinois Natural History Department of Plant Biology, University of Illinois at Urbana-Champaign, Room 265 Morrill Hall, 505 South Goodwin Avenue, Urban Avenue, Hall, 505 South Goodwin Illinois at Urbana-Champaign, Room 265 Morrill University of Department of Plant Biology, of Cusco, Junín, and Madre de Dios yielded specimens of Cancellidium applanatum of Cusco, Junín, and Abstract: Sporoschisma juvenile unsequenced species: Cancellidium data was generated for three previously records for Perú. Molecular applanatum, neotropics with an accompanying phylogeny based on partial 28S nuclear ribosomal large-subunit sequence neotropics with an accompanying phylogeny of The sexual morph data. USA; corresponding author e-mail: [email protected] USA; corresponding author © 2014 International Mycological Association © 2014 International Mycological conditions: distribute and transmit the work, under the following are free to share - to copy, You Attribution: 2 3 1 Steven E. Zelski Steven E. Phylogeny and morphology of dematiaceous freshwater microfungi from microfungi freshwater dematiaceous of and morphology Phylogeny Perú Non-commercial: No derivative works: Any of the above conditions can be waived if you get For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at http://creativecommons.org/licenses/by-nc-nd/3.0/legalcode. Nothing in this license impairs or restricts the author’s moral rights. permission from the copyright holder. time, resulting in the new combination 10 December 2014. Accepted: 24 November 2014; Published: Article info: Submitted: 20 January 2014; cultural studies. Molecular data from ascospores and conidia of cultural studies. Molecular data from ascospores that a connection to phylogenetically related to collections of and S. uniseptatum). originally collected from submerged wood blocks of Ochroma pyramidale & been reported from many Paleotropical localities (Webster Davey 1980, Shaw 1994, Ho et al. 2001, Sivichai et al. 2002, Fryar Zhao recovered from low and middle altitudes along the elevational Yeung gradient, but not from high altitude aquatic habitats. et al. (2006) suggested that the congeneric INTRODUCTION submerged, During a study of ascomycetes colonizing debris in freshwater decomposing woody and herbaceous Perú extending from habitats along an elevational gradient in Andes (2010–2012), the Peruvian Amazon to the Peruvian were encountered. numerous freshwater mitosporic fungi Shearer into three ecological groups: (1) freshwater hyphomycetes; into three ecological groups: (1) freshwater (3) freshwater (2) aeroaquatic hyphomycetes; and This study deals with miscellaneous mitosporic ascomycetes. one aeroaquatic hyphomycete (Cancellidium applanatum and four species of miscellaneous mitosporic ascomycetes (Cordana abramovii, doi:10.5598/imafungus.2014.05.02.07 VOLUME 5 · NO. 2 Zelski et al.
has brown 5-septate, cylindrical, verruculose conidia; and phase optics. Digital micrographs were obtained with the S. parcicuneatum has brown, 1(–3)-septate, cuneiform, SPOT Insight 12 Mp colour camera and Spot Advanced verruculose conidia (Goh et al. 1997). software. Images were processed with Adobe Photoshop and Sexual reproductive structures of Melanochaeta assembled with Adobe InDesign. hemipsila were found among conidiophores of S. saccardoi For single spore isolation, sterile dissecting needles were
ARTICLE from substrates collected in Cusco and Junin. Melanochaeta used to spread ascospores or conidia on antibiotic water hemipsila has been connected to S. saccardoi based on agar (AWA]: 20 g agar (Difco), 0.5 g streptomycin sulfate,
studies in which the asexual morph was produced from 0.5 g penicillin G (Sigma) and 1000 mL deionized H2O. colonies derived from ascospores (Müller et al. 1969, Nag Raj Single germinated ascospores or conidia were transferred to 1975, Sivichai et al. 2000). The sexual morph of the Peruvian PYG+Ab agar plates: 1.25 g peptone, 1.25 g yeast extract, ! 18 g agar (Difco), 5 g D-glucose (Acros), 0.5 g streptomycin brown to black ascomata with short conical beaks; numerous, sulfate, 0.5 g penicillin G (Sigma), and 1000 mL deionized
septate, capitate setae arising from the external ascomal H2O. They were then grown at ambient temperature with wall; clavate, unitunicate, 8-spored asci with an I- refractive 12/12 hr light/dark conditions. apical apparatus; biseriate, cylindrical to curved, 5-septate ascospores with olivaceous to brown central cells, hyaline end cells, lacking sheaths or appendages. DNA extraction was performed on mycelium scraped with The goals of this study were to: (1) describe, illustrate, and a sterile spatula from PYG+Ab agar plates. Mycelium was provide voucher specimens and sequences for the foregoing * species of freshwater mitosporic fungi for which pure cultures mortar and pestle and DNA was extracted with a DNeasy were obtained; (2) compare and contrast these fungi with Plant Mini Kit (Qiagen Sciences, Valencia, CA) according to morphologically similar and genetically related taxa; and (3) the manufacturer’s instructions. PCR of extracted DNA was construct a molecular phylogeny using 28S large subunit performed using Illustra Ready-To-Go™ PCR Beads (GE (LSU) nrDNA to elucidate the evolutionary relationships of Healthcare) using the primer pair LROR and LR6 (Rehner & these fungi with other Ascomycota. Samuels 1994, Vilgalys & Hester 1990) on an MJ Research PTC-200 thermocycler using the following parameters: initial denaturation at 95 °C for 5 min, followed by 40 cycles at 95 °C MATERIALS AND METHODS for 30 s, 50 °C for 15 s, 72 °<