During a biodiversity study of micromycetes occurring on bark and decorticated wood of a native gymnosperm, Podocarpus parlatorei Pilg., from Argentina, new taxa of Crassoascus and Iodosphaeria were found. They are described, illustrated and compared with the known species within each genus

Catania M del V

Romero AI.

mycosphere

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Mycosphere Doi 10.5943/mycosphere/3/1/4 37 Crassoascus  monocaudatus  and  Iodosphaeria  podocarpi,  two  new  species  on Podocarpus parlatorei from “Las Yungas”, Argentina   Catania M del V1 and Romero AI2  1Fundación  Miguel  Lillo,  Laboratorio  de  Micología.  Miguel  Lillo  251.  San  Miguel  de  Tucumán  (4000),  Argentina. mailto:mcatania@tucbbs.com.ar 2PRHIDEB-CONICET,  Departamento  de  Biodiversidad  y  Biología  Experimental,  Facultad  de  Ciencias  Exactas  y Naturales,  Universidad  de  Buenos  Aires,  Ciudad  Universitaria,  Pabellón  2  4to  piso  (EHA1428),  Buenos  Aires, Argentina. romero@bg.fcen.uba.ar  Catania M del V, Romero AI. 2012 – Crassoascus monocaudatus and Iodosphaeria podocarpi, two new species on Podocarpus parlatorei from “Las Yungas”, Argentina. Mycosphere 3(1), 37-44, Doi 10.5943/mycosphere/3/1/4  During a biodiversity study of micromycetes occurring on bark and decorticated wood of a native gymnosperm,  Podocarpus  parlatorei  Pilg.,  from  Argentina,  new  taxa  of  Crassoascus  and Iodosphaeria  were  found.  They  are  described,  illustrated  and  compared  with  the  known  species within each genus.  Key words – Amphisphaeriaceae – Ascomycota – Clypeosphaeriaceae – Iodosphaeriaceae  Article Information  Received 12 January 2012 Accepted 16 January 2012 Published online 25 January 2012 *Corresponding author: Catania M del V – e-mail – mcatania@tucbbs.com.ar   Introduction During a biodiversity study of micromy-cetes occurring on bark and decorticated wood of a native gymnosperm, Podocarpus parlatorei Pilg., from Argentina, two undescribed ascomy-cetes were found. One fits the genus Crassoas-cus  Barrasa,  Checa  &  A.T.  Martínez  and  the other the genus Iodosphaeria Samuels, E. Müll. & Petrini. Crassoascus is characterised by subcorti-colous,  erumpent,  black  ascomata,  separate or gregarious on the host surface; apex papillate, ostiolate. The asci are cylindrical, unitunicate, thick-walled, apical ring enlarged, amyloid and the ascospores are brown, fusoid, asymmetric, septate,  gutulate  when  young,  smooth-walled, apiculate at each end with a conspicuous termi-nal pore and hyaline refractive cap-like appen-dages.  There  are  no  data  about  its  anamorph. Two species have been described (www.index-fungorum.org). Iodosphaeria is characterised by superfi-cial,  black  ascomata,  with  unbranched,  brown flexuous  hairs  radiating  from  the  peridium surface.  The  asci  are  unitunicate  with  an amyloid,  discoid  apical  ring  which  may  be lacking,  and  ascospores  are  allantoid  to  ellip-soidal, aseptate and hyaline, with or without a sheath.  Synanamorphs  in  Selenosporella  G. Arnaud  ex  MacGarvie  and  Ceratosporium Schwein. have been  found  in association with species of Iodosphaeria (Samuels et al. 1987). Iodosphaeria  is  a  genus  comprising  seven species (www.indexfungorum.org). The two genera are reported for the first time on gymnosperms and from Argentina, and in the case of Crassoascus this is also the first record for South America. For Iodosphaeria, a key is provided for the known species. Both of these  fungi  are  morphologically  distinct  when compared to the known species.  Mycosphere Doi 10.5943/mycosphere/3/1/4  38 Methods  Isolates and morphology The fungi were found on dead branches of  Podocarpus  parlatorei,  collected  in  the forest of Sierra de Medina (Dep. Burruyacu) in Tucumán province, Argentina. The studied area belongs to the District Mountain Forest of the phytogeographic  region  of  “Las  Yungas”,  a subtropical  zone  of  the  Amazonic  Domain,  a Neotropical  Region  which  covers  the  Andes from Venezuela and Colombia, into the north-west of Argentina  as a  narrow fringe through Jujuy, Salta, Tucuman and Catamarca  Provin-ces  (Cabrera  &  Willink  1973,  Hueck  1978). The  material  was  dried  and  deposited  in  the fungal  reference  collection  of  Fundación Miguel Lillo (LIL). Observations, photography and  measurements  of  ascomata,  asci  and ascospores  were  made  on  material  squash-mounted in distilled water, 5% KOH, phloxine and  Melzer‘s  reagent  for  optical  microscopy. The drawings were made using a camera lucida. A  cirrus  of  Crassoascus  was  removed  with  a sterile  needle  and  transferred  to  Petri  dishes containing  PDA  (Hawksworth  et  al.  1995). These were  incubated under  laboratory condi-tions  at  approximately  12  hr  fluorescent  light per day at 25º C. In addition, holotype speci-mens of Crassoascus fusisporus and C. canad-ensis from MA and DAOM were studied.  Results   Crassoascus  monocaudatus  Catania  &  A.I. Romero, sp. nov. Figs 1–7; 8–16 Etymology  –  referring  to  presence  of  a single appendage in the ascospore. A  Crassoascus  monocaudatus  differt  ob ascosporas 58.5–82 x 14–16 μm, 5–8 septatas, et appendice singulari basali quasque indutas. Ascomata  immersed,  subcortical,  erum-pent,  globose  to  suglobose,  0.4–1.25  x  0.4–1 mm, ostiolate, with short neck, neck 0.1–0.25 x 0.1 mm, solitary or gregarious. Peridium 26–45 μm wide, outer layer of thick-walled cells, dark brown, irregular to compressed laterally; inner layer  of  compressed  cells,  with  thin,  hyaline walls. Asci cylindrical apically thickened, with 8-spores,  biseriate,  255–324  x  24–30  μm, stipitate, pedicel 33–45 μm long, subapical ring conspicuous staining blue in Melzer´s reagent, 2–4 x 5–7 μm. Paraphyses abundant, thin, with numerous  guttules,  hyaline,  1.5-2.5  μm  diam. Ascospores  fusiform,  slightly  curved,  (56–)58.5–82(–101) x (13–)14–16(–19) μm, at first hyaline and unicellular with 1-2 large guttules and a basal appendage, later multiseptate (5–)6–8(–9) septa, not constricted at the septa, smooth, bright brown to dark brown with hyaline apical cell and basal appendage, 15.5–27 μm long; the basal appendage collapsed when mature. Anamorph – Not seen. Known  distribution  –  only  from  type locality. Holotype  –  Argentina,  Tucumán,  Burru-yacu, Sierra de Medina, provincial route 310, at 31 Km from Villa Padre Monti, Aguas Negras, Finca Mansilla, on dead branch of Podocarpus parlatorei,  26º22'06''S,  65º03'46'W,  25  Feb 1999, Catania 1241 (Holotype, LIL). Additional  material  examined  –  Cras-soascus  monocaudatus.  Argentina,  Tucumán, Burruyacu, Sierra de Medina, provincial route 310, at 31 Km from Villa Padre Monti, Aguas Negras, Finca Mansilla, on dead branch of Po-docarpus parlatorei, 26º22'06''S, 65º03'46'W, 2 Mar  2000,  Catania  1824  (LIL).  Crassoascus canadensis. Canada, British Columbia, Sidney, beach off Glenelg, 4 October 1991, M.E. Barr 7787  (holotype,  DAOM).  Crassoascus fusisporus.  Spain,  Cáceres,  Natural  Park  of Monfragüe,  Puerto  de  la  Cañadilla,  on  dead branches  of  Erica  arborea,  21  June  1989, Checa,  Barrasa  &  Blanco  (holotype,  MA-26331). Notes – Crassoascus was established by Barrasa et al. (1993) based on the type species C. fusisporus Checa, Barrasa & A.T. Martínez. A second species, C. canadensis M.E. Barr, was proposed  by  Barr  (1994).  Crassoascus  mono-caudatus  is  included  within  the  genus  princi-pally  because  it  shares  with  the  other  two species  an  ascomatal  peridium  with  “textura globulosa”,  asci  with  a  prominent  amyloid subapical  ring  and  brown,  fusiform,  septate ascospores. Crassoascus monocaudatus is dist-inguished  by  ascospore  features.  They  are longer  than  the  ascopores  of  the  other  two species,  have  more  septa  and  a  long  basal appendage. Holotypes of the other two species were  studied.  Crassoascus  fusisporus  has Mycosphere Doi 10.5943/mycosphere/3/1/4             39 Figs 1–7 – Crassoascus monocaudatus. (holotype). 1, 2 Ascomata on substratum. 3 Ascus. 4 Apex of ascus, with amyloid apical ring. 5 Young ascospores with basal appendage. 6 Mature ascospore with  appendage.  7  Ascospores  without  appendages.  Bars:  1–2=  0.5  mm;  3–7=  10  µm. smaller ascospores (40–45 x 8–10 μm), with 3–5 septa, apiculate at both ends with a conspi-cuous terminal pore and hyaline cap-like appen-dages  and  C.  canadensis  has  symmetrical ascospores, 26–38 x 8–12 μm, with 3–7 septa and a slight constriction at the median septum. Originally,  Crassoascus  was  placed within  Clypeosphaeriaceae  by  Barrasa  et  al. (1993),  later  Barr  (1994)  transferred  it  to  the Amphisphaeriaceae. Kang et al. (1999) studied the holotype of C. fusisporus and suggested to place it within the Clypeosphaeriaceae despite the “textura globulosa” of the peridium and the atypical arrangement of the ascospores. Accor-ding to Eriksson (2006) the genus is maintained in  the  Clypeosphaeriaceae.  Huhndorf  (pers. comm.) has pointed out that the Amphisphae-riaceae  has  not  yet  been  adequately  separated by molecular data from the Clypeosphaeriaceae but  if  the  two  families  were  united, Amphisphaeriaceae  is  the  older  name.  The placement of Crassoascus  into the Annulatas-Mycosphere Doi 10.5943/mycosphere/3/1/4  40 caceae by Kirk et al. (2008) is not correct. This is the first time that a species of the genus has been reported from Argentina. It is probably  also  the  first  record  from  South America.  It  is  also  the  first  account  on  a gymnosperm. Crassoascus fusisporus is repor-ted as a saprobe on wood on dead branches of Erica arborea L. from Spain and C. canadensis on  wood  of  Salix  (holotype);  on  wood  or periderm of woody plants (Acer macrophyllum Pursh;  Cornus  occidentalis  Coville)  from Canada.  These  two  species  from  the  northern hemisphere were found in areas with a Mediter-ranean  climate,  however  C.  fusisporus  from Spain  was  collected  at  the  end  of  summer-beginning of spring where the summer has high temperatures  and  the  spring  is  rainy.  The Argentine  collections  were  also  found  in summer  although  in  a  subtropical  area.  The Canadian species was found in autumn but in a place  where  the  daily  temperatures  seldom climb above 31°C. Isolations  were  attempted  from  ascos-pores of the Argentine specimen but unfortuna-tely C. monocaudatus did not grow in culture. Barrasa  et  al.  (1993)  obtained  an  Alternaria anamorph  in pure culture, but the relationship with  C.  fusiporus  remains  uncertain  and  the chance of a contamination cannot be rejected.  Iodosphaeria  podocarpi  Catania  &  A.I. Romero, sp. nov. Figs 17-25 Etymology  –  referring  to  the  associated plant, Podocarpus. A  Iodosphaeria  phyllophila  differt  quod longitudinis respectu minores habe ascosporas 19-20 (-22) µm. Ascomata  globose  to  subglobose,  220–250 x 220–240 µm, superficial black, surround-ded by abundant, dark brown, septate, flexuous, unbranched  hairs,  7–8  µm  wide,  to  500  µm long; ostiole apical, periphysate. Peridium 40–80  µm  wide,  comprising  outer  layers  of  dark brown cells forming a “textura angularis” and inner layers of flattened light brown to hyaline cells.  Paraphyses  up  to  4  µm  wide,  hyaline, filamentous, septate, flexuous, deliquescing and disappearing  when  mature.  Asci,  cylindrical, unitunicate,  97.5–120  x  8–12  µm  8-spored, uniseriate  or  partially  biseriate,  short  pedice-llate,  apex  rounded,  with  an  amyloid  apical ring.  Ascospores  inequilaterally  ellipsoid  to navicular, slightly allantoid, often acute at both ends, unicellular, hyaline, smooth, 19–20(–22) x 4.5–5(–6.5) µm. Anamorph:  Selenosporella-like,  found near or among the ascomata in nature. Conidio-phores 100–140(–250) x 4–5 µm, arising from repent, coarse, brown hyphae; erect, straight or flexuous, branched, gregarious, septate, brown, smooth. Conidiogenous cells polyblastic, 13–18 x  4  µm,  integrated  and  terminal,  tapering slighly from base to tip, with minute denticles on the tip of each conidiogenous cell. Conidia straight  or  slightly  curved,  aseptate,  hyaline, 14–17 x 1 µm. Known  distribution  –  only  from  type locality. Holotype  –  Argentina,  Tucumán,  Burru-yacu, Sierra de Medina, provincial route 310, at 31 Km from Villa Padre Monti, Aguas Negras, Finca  Mansilla,  on  branch  of  Podocarpus parlatorei,  26º22'06''S,  65º03'46'W,  1  April 1998, Catania 773 (Holotype, LIL). Notes  –  Iodosphaeria  podocarpi resembles I. polygoni W.H. Hsieh, C.Y. Chen & Sivan. and I. phyllophila (Mouton) Samuels, E.  Müll.  &  Petrini  because  of  the  positive reaction in Melzer´s reagent of the apical ring. I.  polygoni  has  ascospores  of  similar  length (18–23 µm long) but I. polygoni has ellipsoidal ascospores  which  are  wider  (5.5–8  µm)  and longer asci (up to 150 µm  long) (Hsieh et al. 1997). The Argentine collection differs from I. phyllophila, which has longer ascospores (16–31  µm  long)  with  both  ends  less  acute  than those of I. podocarpi (Samuels et al. 1987). This  is  the  first  report  of  the  genus Iodosphaeria  on  a  gymnosperm.  Previously recorded  substrata  were  arborescent  ferns  and dicots for I. phyllophila and various monocots for other species. Species of the genus  Iodos-phaeria  have  been  reported  from  tropical  and temperate regions of the world: China (Taiwan, Hong  Kong);  USA  (Louisiana);  Europe (Belgium,  Germany,  Great  Britain);  New Zealand  and  South  America  (Brazil,  French Guiana) (Samuels et al. 1987, Barr 1993, Hyde 1995, Candoussau et al. 1996, Hsieh et al. 1997, Taylor & Hyde 1999). The taxonomic position of this genus is still unclear and it has been placed in variousMycosphere Doi 10.5943/mycosphere/3/1/4  41   Figs  8–16  –  Crassoascus  monocaudatus  (holotype).  8,  9  Ascomata on  substratum. 10  Asci  and ascospores.  11  Ascus  with  ascospores.  12,  13  Amyloid  apical  ring.  14  Young  ascospore  with appendage (arrow). 15 Mature ascospore with appendage (arrow). 16 Matures ascospores. Bars: 8–9 = 0.5 mm; 10 = 50 µm; 11–16 = 10 µm.  families  by  different  authors.  Samuels  et  al. (1987)placed Iodosphaeria in the Amphisphae-riaceae,  Barr  (1990,  1994)  suggested  that  the genus  Iodosphaeria  is  better  placed  in  the Lasiosphaeriaceae  (Sordariales),  Réblová (1999) placed it in the Trichosphaeriaceae and Eriksson et al. (2001) placed it in the Amphi-sphaeriaceae. It was placed in its own family, Iodosphaeriaceae  by  Hilber  &  Hilber  (2002).Mycosphere Doi 10.5943/mycosphere/3/1/4  42    Figs. 17–25 – Iodosphaeria podocarpi. 17, 18 Ascomata on substrata. 19 Ascoma. 20 Peridium. 21 Unbranched hairs arising from peridium. 22 Asci. 23 Apex of ascus, with an amyloid apical ring. 24 Ascus with apical ring and ascospores. 25 Selenosporella conidiophores. Bars: 17 = 750 µm; 18 = 250 µm; 19 = 75 µm; 20–21= 25 µm; 22–23 = 10 µm; 24 = 5 µm; 25 = 10 µm.  Mycosphere Doi 10.5943/mycosphere/3/1/4  43 Key to Iodosphaeria species known to date 1. Asci with apical apparatus .............................................................................................................. 2 1. Asci lacking apical apparatus ......................................................................................................... 6  2. Apical apparatus J+ ........................................................................................................................ 3 2. Apical apparatus J- ......................................................................................................................... 5 3. Asci longer than 150 µm, ascospore ellipsoidal, 18–23 x 5,5–8 µm ............................... I. polygoni 3. Asci less than 150 µm in length ................................................................…….………………….4 4. Ascospores allantoid, commonly on debris of tree ferns, Cyperaceae and on fallen, dead twigs of Acer, Salix and Populus............................................................................................. I. phyllophila 4.  Ascospores  inequilaterally  ellipsoid  to  navicular,  on  fallen,  dead  twigs  of Podocarpaceae………….……………………………………………………………...... I. podocarpi 5. Ascospore wall verruculose ..................................................................................... I. arundinariae 5. Ascospore wall smooth ......................................................................................................... I. tarda 6. Ascospores ellipsoidal, with a mucilaginous sheath, 19–29 x 7–9 µm ............................ I. ripogoni 6.  Ascospores  ellipsoidal  to  fusiform,  lacking  a  mucilaginous  sheath,  14–22  x  4–6  µm .................................................................................................................................... I. hongkongensis  Acknowledgements We  thank  the  directors of  DAOM  and MA  fungal  reference  collections  for  kindly providing  material  for  investigation.  We  are also  grateful  to  Dr.  Dave  Minter  for  his  help with the manuscript. This is publication N° 189 of  the  PRHIDEB-PROPLAME,  partially financed  by  the  Argentine  National  Research Council.  References  Barr ME. 1990 – Prodromus to non-lichenized pyrenomycetous  members  of  the  class Hymenoascomycetes.  Mycotaxon  39, 43-184. Barr  ME.  1993  –  Redisposition  of  some  taxa described by J.B. Ellis. Mycotaxon 46, 45–76. Barr  ME.  1994  –  Notes  on  the Amphisphaeriaceae and related families. Mycotaxon 51, 191–224. Barrasa  JM,  Checa  J,  Martínez  AT.  1993  – Crassoascus, a new nonstromatic genus in  the  Clypeosphaeriaceae.  Mycotaxon 46, 299–305. Cabrera AL, Willink A. 1973 – Biogeografía de América  Latina.  Monografía  Nº  13. Serie de Biología. Secretaria Gral. de la OEA. 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Crassoascus monocaudatus and Iodosphaeria podocarpi, two new species on Podocarpus parlatorei from

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