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

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Houbraken et al. ARTICLE Table 1. Overview of Penicillium strains used in this study. CBS no. Other no. Name Substrate, locality 449.78 IBT 21509=IBT 3473=IBT 6753 P. carneum Cheddar cheese 466.95 ATCC 46837=IBT 6885 P. carneum Cured meat, Germany 467.95 IBT 3466 P. carneum Hotwater tank, North Sealand, Denmark 112297 T IBT 6884 P. carneum Type, mouldy rye bread, Denmark 463.95 IBT 12392 P. paneum Chocolate sauce, Norway 464.95 IBT 11839 P. paneum Rye bread (non preserved), Odense, Denmark 465.95 IBT 13929 P. paneum Mouldy baker’s yeast, Vangede, Denmark 101032 T IBT 21541=IBT 12407 P. paneum Type, mouldy rye bread, Denmark 112296 IBT 21729 P. paneum Cassava chips, Africa 112294 IBT 16402=NRRL 1168 P. paneum Unknown substrate, Ottawa , Canada 128137 T DTO 70G9 = IBT 29551 P. psychrosexualis Type, wooden crate in cold-store of apples, the Netherlands 128136 DTO 70H7 P. psychrosexualis Wooden crate in cold-store of apples, the Netherlands 128035 DTO 70H4 P. psychrosexualis Wooden crate in cold-store of apples, the Netherlands 128036 DTO 70H9 P. psychrosexualis Wooden crate in cold-store of apples, the Netherlands 135.67 IBT 19475=MUCL 8491 P. roqueforti Blue veined cheese, Germany 221.30 NT ATCC 10110=ATCC 1129=CECT P. roqueforti Neotype, French Roquefort cheese, USA 2905=IBT 6754=IFO 5459=IMI 024313=NRRL 849 234.38 IBT 19781=IMI 291202 P. roqueforti Blue Cheshire cheese 479.84 IBT 21543 P. roqueforti Mouldy baker’s yeast, Denmark 498.73 ATCC 24720=FRR 1480=IBT 19476=IMI 174718=IMI 291199=VKM F-1748 P. roqueforti Fruit of Malus sylvestris (apple), Russia the manufacturer’s instructions. The ITS regions (ITS), a part of the β-tubulin (BenA) or calmodulin (Cmd) gene were amplified and sequenced according the method described in Houbraken et al. (2007). Each dataset was aligned using the Clustal W program in MEGA5 (Tamura et al. 2007), and subsequently manually optimised. The evolutionary history was inferred by using the Maximum Likelihood (ML) method based on the Tamura-Nei model (Tamura & Nei 1993). The bootstrap consensus tree inferred from 1 000 replicates is taken to represent the evolutionary history of the taxa analysed (Tamura et al. 2007). The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1 000 replicates) is shown next to the branches. Initial tree(s) for the heuristic search were obtained automatically as follows. When the number of common sites is < 100 or less than one fourth of the total number of sites, the maximum parsimony method was used; otherwise BIONJ method with MCL distance matrix was used. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. All positions containing gaps and missing data were eliminated. Evolutionary analyses were conducted in MEGA5 (Felsenstein 1985, Tamura et al. 2007). All phylograms were rooted with Penicillium egyptiacum CBS 244.32 NT . The newly obtained sequences were deposited in GenBank under accession numbers HQ442319–HQ442359. Extrolite analysis Plugs with mycelium and agar were extracted from 7 d old cultures grown on CYA and YES. Extracts were prepared using the method described by Smedsgaard (1997). Each extract was filtrated through a 0.45 PTFE filter and subsequently analysed using HPLC with diode array detection (DAD) detection. The UV spectrum and the RI value, and comparison with authentic chemical standards, were used to characterise the extrolites produced (Frisvad & Thrane 1987). Results Phylogeny The ITS regions and parts of the β-tubulin (BenA) and calmodulin (Cmd) gene were sequenced and analysed. The BenA alignment included 432 positions, and 35 positions were parsimony informative. The bootstrap consensus tree based on the results of the maximum likelihood analysis of this alignment is shown in Fig. 1. The total length of the calmodulin alignment was 500 positions long, and 27 sites were parsimony informative. The bootstrap consensus tree derived from the maximum likelihood analysis is shown in Fig. 2. The phylogram in Fig. 3 is based on the ITS regions (incl 5.8S rDNA), and 585 bases were used in the maximum likelihood analysis. Of these 585 characters, 16 were parsimony informative (including alignment gaps). The result of the analysis of the three datasets shows that P. psychrosexualis belongs to the series Roqueforti. The species is related to P. carneum and P. roqueforti in all three analysed loci, and P. paneum is basal to these three species. Penicillium carneum is the closest relative of P. psychrosexualis in the tubulin phylogram (99 %, Fig. 1), and P. roqueforti is basal to these two species. However, this relationship is not supported in the phylograms based on the calmodulin and ITS sequences. In these datasets, P. carneum and P. roqueforti are 172 i m a f U N G U S
Sex in Penicillium series Roqueforti 93 86 CBS 467.95 P. carneum CBS 112297 T P. carneum CBS 466.95 P. carneum CBS 128137 T P. psychrosexualis ARTICLE 87 CBS 128136 P. psychrosexualis CBS 128035 P .psychrosexualis CBS 449.78 P .carneum CBS 234.38 P. roqueforti 49 CBS 498.73 P. roqueforti CBS 135.67 P. roqueforti 85 CBS 479.84 P. roqueforti CBS 112294 P. paneum CBS 101032 T P. paneum CBS 221.30 NT P. roqueforti 0.01 92 CBS 464.95 P. paneum CBS 463.95 P. paneum CBS 112296 P. paneum CBS 244.32 P. egyptiacum Fig. 1. Bootstrap consensus tree from a maximum likelihood analysis of partial β-tubulin sequences. The bootstrap values from 1 000 replicates are shown at the nodes, the branches in bold have a bootstrap support higher than 95 %. The tree was rooted with Penicillium egyptiacum CBS 244.32 NT . CBS 449.78 P. carneum 64 CBS 112297 T P. carneum CBS 466.95 P. carneum CBS 467.95 P. carneum CBS 479.84 P. roqueforti CBS 135.67 P. roqueforti 77 69 63 CBS 498.73 P. roqueforti CBS 221.30 NT P. roqueforti CBS 128035 P. psychrosexualis CBS128136 P. psychrosexualis CBS 128137 T P. psychrosexualis CBS 101032 T P. paneum 94 CBS 465.95 P. paneum CBS 464.95 P. paneum 0.01 CBS 112294 P. paneum CBS 112296 P. paneum CBS 244.32 NT P.egyptiacum Fig. 2. Bootstrap consensus tree from a maximum likelihood analysis of partial calmodulin sequences. The bootstrap values from 1 000 replicates are shown at the nodes, the branches in bold have a bootstrap support higher than 95 %. The tree was rooted with Penicillium egyptiacum CBS 244.32 NT . v o l u m e 1 · n o . 2 173
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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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vier’s favour was that they publi
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Welcome to the pressure dome: inves
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REPORTS IMC9: Moments to reminisce
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IMA Medals awarded at IMC9 The high
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Society of America, in the form of
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ecognition based on several criteri
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8. Preparing agreed responses to In
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Physiological differences between w
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Trizodia acrobia, hyphae enveloping
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International Society for Fungal Co
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steering committee mentioned earlie
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Iranian Mycological Society It give
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The Forayer, and courses was missed
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accounts of studies of particular g
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62 nd Annual Applied Microscopy Con
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IMA Fungus · volume 1 · no 2: 101
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Calonectria lauri sp. nov. Table 1.
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Calonectria lauri sp. nov. ARTICLE
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Calonectria lauri sp. nov. Brayford
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How to describe a new fungal specie
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Johansonia al. 1990) and LSU1Fd (Cr
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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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