Volume 1 Â· No. 2 Â· December 2010 V o lu m e 1 Â· N o ... - IMA Fungus

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Varga et al. ARTICLE Table 1. Isolates of Aspergillus and Emericella spp. examined in this study. Species Strain No. a Substratum, country, location GenBank No. β-tubulin calmodulin ITS A. aeneus CBS 128.54 T = NRRL 4769 Forest soil, Modilen, Somalia EF652298 EF652386 EF652474 A. crustosus CBS 478.65 T = NRRL 4988 Skin scrapings, man, Kankakee, Illinois, USA EF652313 EF652401 EF652489 A. eburneocremeus CBS 130.54 T = NRRL 4773 Forest soil, Modien Forest, Somalia EF652300 EF652388 EF652476 A. heyangensis CBS 101751 T Placentae of Gossypium sp., Heyang, Shaanxi FJ491520 FJ491521 FJ491522 Province, China A. karnatakaensis CBS 102800 T = IBT 22153 Soil under coconut palm in coffee plantation, EU482438 EU482431 EU482441 India, Karnataka A. karnatakaensis CBS 102799 = IBT 22154 Soil under coconut palm in coffee plantation, EU482436 EU482430 EU482443 India, Karnataka A. karnatakaensis NRRL 4649 Soil in the Machrar river bed located in district EF652292 EF652380 EF652468 Bansa, state Madhya Pradesh, India E. bicolor CBS 425.77 T Soil from Artemisia grassland, USA, Wyoming, EF652335 EF652423 EF652511 Teton Basin E. discophora CBS 469.88 T = IBT 21910 Soil, Spain AY339999 EU443970 EU448272 E. discophora CBS 470.88 = IBT 21911 Forest soil, Spain AY340000 EU443969 EU448266 E. foeniculicola CBS 156.80 T Foeniculum vulgare seed, China EU443990 EU443968 EU448274 E. heterothallica CBS 489.65 T Soil, Costa Rica EU076369 EU076361 AB248987 E. spectabilis CBS 429.77 T Coal mine spoil material, Wyoming, USA, Seminole no. 1 mine EU482437 EU482429 EU482442 a Cultures are deposited in/were obtained from the following collections: CBS, CBS-KNAW Fungal Biodiversity Centre, Utrecht, the Netherlands; IBT, Culture Collection of Fungi, Mycology Group, BioCentrum-DTU, Technical University of Denmark, Lyngby, Denmark; NRRL, Agricultural Research Service Culture Collection, Peoria, IL, USA. of each agar medium were taken and pooled together into the same vial for extraction with 0.75 mL of a mixture of ethyl acetate/dichloromethane/methanol (3:2:1) (v/v/v) with 1 % (v/v) formic acid. The extracts were filtered and analyzed by HPLC using alkylphenone retention indices and diode array UV-VIS detection as described by Frisvad & Thrane (1987, 1993), with minor modifications as described by Smedsgaard (1997). The column used was a 50 × 2 mm Luna C-18 (II) reversed phase column (Phenomenex, CA, USA) fitted with a 2 × 2 mm guard column. Genotypic analysis The cultures used for the molecular studies were grown on malt peptone (MP) broth using 1 % (w/v) of malt extract (Brix 10) and 0.1 % (w/v) bacto peptone (Difco), 2 mL of medium in 15 mL tubes. The cultures were incubated at 25 °C for 7 d. DNA was extracted from the cells using the Masterpure yeast DNA purification kit (Epicentre Biotechnology.) according to the instructions of the manufacturer. The ITS region and parts of the b-tubulin and calmodulin genes were amplified and sequenced as described previously (Varga et al. 2007a–c). Data analysis The sequence data was optimised using the software package Seqman from DNAStar Inc. Sequence alignments were performed by MEGA v. 4.0 (Tamura et al. 2007) and improved manually. For parsimony analysis, PAUP v. 4.0b10 software was used (Swofford 2003). Alignment gaps were treated as a fifth character state and all characters were unordered and of equal weight. Maximum parsimony analysis was performed for all data sets individually using the heuristic search option with 100 random taxa additions and tree bisection and reconstruction (TBR) as the branch-swapping algorithm. Branches of zero length were collapsed and all multiple, equally parsimonious trees were saved. The robustness of the trees obtained was evaluated by 1000 bootstrap replications (Hillis & Bull 1993). Eurotium heterothallica was used as outgroup in these analyses (Houbraken et al. 2007). The alignments were deposited in TreeBASE () under accession number S11027. RESULTS AND DISCUSSION Phylogeny Of the aligned β-tubulin sequences, a portion with 438 positions, including 107 parsimony informative characters, was selected for the analysis; MP analysis of the sequence data resulted in two similar, equally most parsimonious trees (tree length = 289 steps, consistency index = 0.7855, retention index = 0.7919), one of which is shown in Fig. 1. The calmodulin data set consisted of 492 characters, including 188 198 i m a f U N G U S
Aspergillus karnatakaensis sp. nov. “A. aeneus” NRRL 4649 A. karnatakaensis CBS 102799 A. karnatakaensis CBS 102800 A. aeneus NRRL 4769 A. aeneus CBS 128.54 ARTICLE E. discophora CBS 469.88 E. discophora CBS 470.88 A. eburneocremeus CBS 130.54 A. eburneocremeus NRRL 4773 E. foeniculicola CBS 156.80 E. bicolor CBS 425.77 E. spectabilis CBS 429.77 A. heyangensis CBS 101751 A. crustosus CBS 478.65 A. crustosus NRRL 4988 E. heterothallica CBS 489.65 Fig. 1. One of the two equally MP trees obtained based on phylogenetic analysis of b-tubulin sequence data of Aspergillus sect. Aenei. Numbers above branches are bootstrap support values. Only values above 70 % are indicated. A. karnatakaensis CBS 102799 A. karnatakaensis CBS 102800 “A. aeneus” NRRL 4649 A. aeneus NRRL 4769 A. aeneus CBS 128.54 E. discophora CBS 469.88 E. discophora CBS 470.88 E. heterothallica CBS 489.65 A. eburneocremeus CBS 130.54 E. bicolor CBS 425.77 E. spectabilis CBS 429.77 E. foeniculicola CBS 156.80 A. heyangensis CBS 101751 A. crustosus CBS 478.65 A. crustosus NRRL 4988 Fig. 2. The single MP tree obtained based on phylogenetic analysis of calmodulin sequence data of Aspergillus sect. Aenei. Numbers above branches are bootstrap support values. Only values above 70 % are indicated. v o l u m e 1 · n o . 2 199
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T h e G l o b a l M y c o l o g i c
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IMA vision My first opportunity to
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undescribed fungal diversity will r
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Fungi on German TV: Focus on black
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REPORTS IMC9 Edinburgh: a selection
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REPORTS IMC9: Moments to reminisce
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IMA Medals awarded at IMC9 The high
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Physiological differences between w
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International Society for Fungal Co
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The Forayer, and courses was missed
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Mycology in Egypt ACKNOWLEDGEMENTS
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Mycology in Egypt Aleem AA (1952b)
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Volume 1 Â· No. 2 Â· December 2010 V o lu m e 1 Â· N o ... - IMA Fungus

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