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The genus Cladosporium and similar dematiaceous ... - CBS - KNAW

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Hormoconis resinae <strong>and</strong> morphologically <strong>similar</strong> taxa<br />

Exsiccati <strong>and</strong> types<br />

Dematium nigrum [scr. Link]. E. Hbr. Link (23) = Sporocybe<br />

resinae III. 341 [scr. ? ] (herb. Link, B).<br />

Hormodendrum resinae Lindau, n. sp. Fl. v. Hamburg 206, auf<br />

Harz an Picea excelsa, Sachsenwald, leg. O. Jaap, 29-4-1906.<br />

[scr. Lindau]. (DAOM 41888, slide prepared from the holotype<br />

preserved in B.)<br />

Pycnostysanus resinae Lindau nov. gen. et nov. spec., Kabát et<br />

Bubák: Fungi imperfecti exsiccati no. 99. Auf erhärteten Fichtenharz<br />

an Brockenweg, am Dreieckigen Pfahl in Harz, Deutschl<strong>and</strong>, leg.<br />

G. Lindau, 13.VIII. 1903 (holotype, B).<br />

Racodium resinae Fries. E. Hbr. Link, Fries legi, Smol. [scr.<br />

Fries]. (DAOM 41890, slide prepared from herb. Link, B). This<br />

is the presumed holotype of R. resinae, the basionym for the<br />

resin fungus, Sorocybe resinae. <strong>The</strong> specimen includes dark,<br />

decapitated synnemata, brown conidia with laterally thickened<br />

walls, <strong>and</strong> acropetal conidial chains, allowing it to be recognised<br />

as the fungus we now know as S. resinae. Fries perhaps sent this<br />

fungus to Link to see if it could be differentiated from Coremium<br />

Link. <strong>The</strong> minimal details, that the fungus was collected by Fries,<br />

presumably in Smål<strong>and</strong> (a province of Sweden), match the details<br />

in the protologue of this species.<br />

Sorocybe resinae. “Fungi Rhenani Fasc. II, 1863, L. Fuckel, no.<br />

129, ad Abietis resinam, raro Hieme, in sylva Hostrichiensi” (as<br />

Myxotrichum resinae Fr., DAOM 55543 ex FH). “Flora Suecica,<br />

2956, Ad resinam piceae, Smål<strong>and</strong>: Femsjö, Prostgaidsshogen,<br />

6 Aug. 1929, leg. J.A. Nannfeldt, s.n.” (as Stysanus resinae (Fr.)<br />

Sacc., DAOM 41891 ex UPS). “Flora Suecica, 4709, Ad resinam<br />

abietinum, Uppl<strong>and</strong>: Bondkysko sin Valsätra, 9 May 1932, leg. J.A.<br />

Nannfeldt” (as Hormodendrum resinae Lindau, DAOM 41889 ex<br />

UPS). “[ on wood scr. Berkeley] J.E. Vize, Hereford 1877” (as Torula<br />

pinophila Fr., DAOM 113425 ex K). “Sydow, Mycotheca germanica,<br />

350. Auf Fichtenharz… am Brockenweg 30.9.1904, leg. P. Sydow”<br />

(DAOM 41893).<br />

Other material examined<br />

Sorocybe resinae. Canada, British Columbia: Burnaby, Central Park, on resin of<br />

Tsuga heterophylla, leg. S. & L. Hughes, 17 Aug. 2000 (DAOM 228572a, 228573a);<br />

Cameron Lake, Cathedral Grove, on Pseudotsuga menziesii, leg. isol. S.J. Hughes,<br />

21 Aug. 1957 (DAOM 56088a). Ladysmith, Ivy Green Park, on resinous exudates,<br />

leg. R.J. B<strong>and</strong>oni no. BC-978, 18 Apr. 1960, det. S.J. Hughes (DAOM 70462).<br />

North Vancouver, Lynn Valley Conservation Area, leg. det. S.J. Hughes, 1 Jul. 1975<br />

(DAOM 139385); North Vancouver, Lynn Valley Conservation Area, on bark of living<br />

conifer (probably Pseudotsuga menziesii), leg. isol. K.A. Seifert no. 1574, 26 May<br />

2002 (single conidium isolate, culture <strong>and</strong> specimen DAOM 239134; ITS GenBank<br />

EU030275, LSU GenBank EU030277); Terrace, near Kalum, on Tsuga heterophylla,<br />

leg. W.G. Ziller no. V-6549, 10 July 1950, det. S.J. Hughes (DAOM 59657); Queen<br />

Charlotte Isl<strong>and</strong>s, east coast of Moresby Isl<strong>and</strong>, north side of Gray Bay, 53°08’ N,<br />

131°47’ W, on Picea sitchensis, leg. I. Brodo, M.J. Schepanek, W.B. Schofield,<br />

28 Sep. 1973, det. S.J. Hughes (DAOM 144757); Queen Charlotte Isl<strong>and</strong>s,<br />

Graham Isl<strong>and</strong>, Tow Hill area, on resin of Picea sitchensis, leg. S.A. Redhead no.<br />

4440, 20 Sep. 1982, det. G.P. White (DAOM 184025); Revelstoke, Wigwam, on<br />

Tsuga heterophylla, leg. W. Ziller V-6567 det. S.J. Hughes, 6 Jun. 1950 (DAOM<br />

59710); Vancouver Isl<strong>and</strong>, Cathedral Grove, Cameron Lake, on Pseudotsuga<br />

menziesii, leg. det. S.J. Hughes, 21 Aug. 1957 (DAOM 56088a); Vancouver Isl<strong>and</strong>,<br />

Caycuse, on resin of Pseudotsuga menziesii, leg. det. S.J. Hughes, 17 Jul. 1972<br />

(DAOM 139355); Vancouver Isl<strong>and</strong>, Lake Cowichan, Honeymoon Bay, on resin<br />

of Pseudotsuga menziesii, leg. J Ginns, det. S.J. Hughes, 29 Oct. 1971 (DAOM<br />

134968); Vancouver Isl<strong>and</strong>, Lake Cowichan, Mesachie Lake Forest Experimental<br />

Station, leg. det. S.J. Hughes, 5 Jul. 1972 (DAOM 139277a, DAOM 139278) <strong>and</strong><br />

6 Jul. 1072 (DAOM 139281). Czechoslovakia, Ještěd near Liberec, leg. det. S.J.<br />

Hughes, on resin of Larix europaea, 10 May 1955 (DAOM 51723). United States,<br />

Oregon: Andrews’ Experimental Forest, Forest Service Rd. no 1553, on resin of<br />

Tsuga heterophylla, leg. det. S.J. Hughes, 10 May 1969 (DAOM 134565); Andrews’<br />

Experimental Forest, Blue River, on resin of conifer, cut wood, leg. det. K.A. Seifert<br />

no. 69, 10 Jul. 1981 (DAOM 228203); Oregon, del Norte Co., J. Smith’s State Park,<br />

on Tsuga heterophylla, leg. det. S.J. Hughes, 11 May 1069 (DAOM 134614); Devil’s<br />

Elbow State Park, Cape Perpetus, on Picea sitchensis, leg. det. S.J. Hughes, 6 May<br />

www.studiesinmycology.org<br />

1969 (DAOM 134615); Linn Co., near Cascadia, on Pseudotsuga menziesii, leg. R.<br />

Fogel, det. S.J. Hughes, 14 May 1969 (DAOM 127885); U.S. Forest Service Rd. no.<br />

126, North fork Cape Creek, on resin of Abies gr<strong>and</strong>is, leg. det. S.J. Hughes, 7 May<br />

1969 (DAOM 134852,134563); Willamette National Forest, McKenzie Bridge Camp<br />

Grounds, leg. det. S.J. Hughes, 10 May 1969 (DAOM 134564). Washington: Kittitas<br />

Co., Wanatchee National Forest, Rocky Run, on Abies nobilis, leg. Field Mycology<br />

Class 1955, 22 Jul. 1955, det. S.J. Hughes, (mononematous synanamorph only,<br />

DAOM 118934 ex WSP 45210, as Helminthosporium sp.); Jefferson Co., Olympic<br />

National Forest, 10 mi Camp, Sec. 17, T26N, R3W, on Pseudotsuga mucronata,<br />

leg. Field Mycology Class, 22 Jul. 1955 (DAOM 113801 ex WSP 45212, as<br />

Helminthosporium); Grays Harbor Co., Twin Harbors Beach State Park, resin of<br />

Picea sitchensis, leg. W.B. & V.G. Cooke, 24 Jul. 1951, det. S.J. Hughes (DAOM<br />

118970 ex WSP 28432).<br />

Amorphotheca resinae. Isolated from jet fuel by P. Emonds (culture, DAOM<br />

170427 = ATCC 22711, ITS GenBank EU030278, LSU GenBank EU030280).<br />

Canada, British Columbia, source unknown, isol. “Mrs. Volkoff”, Jul. 1969 (culture,<br />

DAOM 194228, ITS GenBank EU030279).<br />

Seifertia azaleae. All on flower buds of Rhododendron spp. Canada, British<br />

Columbia: Burnaby, Central Park, leg. S.J. Hughes, 17 Aug. 2000 (DAOM 228571);<br />

Vancouver, Stanley Park, leg. K.A. Seifert no. 1571, 11 May 2002 (culture <strong>and</strong><br />

specimen, DAOM 239136, LSU GenBank EU030276). Irel<strong>and</strong>, Munter, Kerry,<br />

near Glenbeigh (ca. N 52° 03’ W 9° 54’), leg. K.A. Seifert no. 3197, 26 Sep. 2006<br />

(culture <strong>and</strong> specimen, DAOM 239135, ITS GenBank EU030273). Netherl<strong>and</strong>s,<br />

Gelderl<strong>and</strong>, Kröller-Müller Museum, leg. K.A. Seifert no. 1235, 12 May 2000 (DAOM<br />

227136). United Kingdom, Wales, Hafod Estate (ca. N 52° 22’ W 3° 51’), leg. K.A.<br />

Seifert no. 3198, 1 Oct. 2006 (culture <strong>and</strong> specimen, DAOM 239137, ITS GenBank<br />

EU030274).<br />

DNA extraction, amplification <strong>and</strong> sequencing<br />

DNA was isolated using a FastDNA Kit <strong>and</strong> the FastPrep<br />

FP120 (BIO 101 Inc.) or an UltraClean Microbial DNA Isolation<br />

Kit (Mo Bio Laboratories, Inc., Solana Beach, CA, U.S.A.) using<br />

mycelium removed from agar cultures. PCR <strong>and</strong> cycle sequencing<br />

reactions were performed on a Techne Genius thermocycler<br />

(Techne Cambridge Ltd.). PCR reactions were performed using<br />

Ready-To-Go Beads (Amersham Canada Ltd.) in 25 µL volumes,<br />

each containing 20–100 ng of genomic DNA, 2.5 units pure Taq<br />

DNA Polymerase, 10 mM Tris-HCl (pH 9.0), 50 mM KCl, 1.5 mM<br />

MgCl 2<br />

, 200 µM of each dNTP, 0.2 µL of each primer (50 µM), <strong>and</strong><br />

stabilizers including bovine serum albumin. <strong>The</strong> reaction profile<br />

included an initial denaturation for 4 min at 94 °C, followed by 30<br />

cycles of 1.5 min denaturation at 95 °C, 1 min annealing at 56 °C,<br />

2 min extension at 72 °C, with a final extension of 10 min at 72 °C.<br />

Amplicons were purified by ethanol/sodium acetate precipitation<br />

<strong>and</strong> resuspended as recommended for processing on an ABI<br />

PRISM 3100 DNA Analyzer or an ABI 373 Stretch DNA Sequencer<br />

(Applied Biosystems, Foster, CA). Amplification products were<br />

sequenced using the BigDye v. 2.0 Terminator Cycle Sequencing<br />

Ready Reaction Kit (ABI Prism/Applied Biosystems) following the<br />

manufacturer’s directions. An approximately 1 000 bp portion of the<br />

large subunit (LSU) ribosomal DNA was amplified <strong>and</strong> sequenced<br />

using primers LR0R <strong>and</strong> LR6, <strong>and</strong> cycle-sequenced using primers<br />

LR0R, LR3R, LR16 <strong>and</strong> LR6 (Vilgalys & Hester 1990, Rehner &<br />

Samuels 1995; www.biology.duke.edu/fungi/mycolab/primers.<br />

htm). <strong>The</strong> complete ITS <strong>and</strong> 5.8S rRNA genes were amplified<br />

<strong>and</strong> sequenced using the primers ITS5 <strong>and</strong> ITS4, with ITS2 <strong>and</strong><br />

ITS3 primers used for cycle sequencing when necessary (White<br />

et al. 1990). Some sequences were derived from single PCR<br />

amplifications of the ITS5–LR6 region.<br />

Data matrices were subjected to parsimony analysis using<br />

heuristic searches in PAUP* v. 4.0b10 (Swofford 2002) with<br />

simple stepwise addition of taxa, <strong>and</strong> tree bisection-reconnection<br />

(TBR) branch swapping. Uninformative characters were removed<br />

for all analyses. Strict consensus trees were calculated, <strong>and</strong> the<br />

robustness of the phylogenies was tested using full bootstrap<br />

analyses (1 000 replications). For all analyses, GenBank accession<br />

numbers are given on the tree figures, <strong>and</strong> the sequences generated<br />

in this study are indicated in bold.<br />

239

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