LABOULBENIALES - Agentschap voor Natuur en Bos

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Table VII: Overview of all sequences in GenBank, found with both the „Laboulbeniales‟ search (first 21sequences) and the „Laboulbeniomycetes‟ search (last 8 sequences). Abbreviations: bp number ofbase pairs, rRNA ribosomal RNA, SSU small subunit, LSU large subunit, TEF1- translation elongationfactor-1 , RPB2 RNA polymerase II second largest subunit.Sequnce lengthSpeciesLocusAccession Publication[bp]Laboulbeniales sp. LM68 18S rRNA 1022 EF060444.1 (a)Rickia passalina 18S rRNA 490 AF432129.1 bCeratomyces mirabilis 18S rRNA 515 AF431764.1 bRhadinomyces pallidus 18S rRNA 515 AF431763.1 bCorethromyces bicolor 18S rRNA 502 AF431762.1 bCorethromyces sp. AW-2001 18S rRNA 515 AF431761.1 bBotryandromyces ornatus 18S rRNA 516 AF431760.1 bStigmatomyces rugosus 18S rRNA 514 AF431759.1 bStigmatomyces scaptomyzae 18S rRNA 518 AF431759.1 bStigmatomyces hydreliae 18S rRNA 516 AF431757.1 bRhachomyces philonthinus 18S rRNA 513 AF431756.1 bZodiomyces vorticellarius SSU rRNA 1083 AF407577.1 cStigmatomyces limnophorae SSU rRNA 1028 AF407576.1 cHesperomyces coccinelloides SSU rRNA 1038 AF407575.1 cHesperomyces virescens LSU rRNA 409 AF298235.1 dStigmatomyces protrudens LSU rRNA 401 AF298234.1 dHesperomyces virescens SSU rRNA 1085 AF298233.1 dStigmatomyces protrudens SSU rRNA 1081 AF298232.1 dPyxidiophora sp. IMI-1989 SSU rRNA 1103 AF313769.1 (e)Kathistes calyculata SSU rRNA 1089 AF313768.1 (e)Kathistes analemnoides SSU rRNA 1096 AF313767.1 (e)Pyxidiophora arvernensis is. AFTOL-ID 2197 TEF-1 678 FJ238412.1 (f)Pyxidiophora arvernensis is. AFTOL-ID 2197 RPB2 857 FJ238377.1 (f)Pyxidiophora arvernensis is. AFTOL-ID 2197 28S rRNA 1080 FJ176894.1 (f)Pyxidiophora arvernensis is. AFTOL-ID 2197 18S rRNA 1478 FJ176839.1 (f)Termitaria snyderi is. DAH 14 18S rRNA 1046 AY212812.1 gLaboulbeniopsis termitarius is. DAH 18 18S rRNA 998 AY212810.1 gPyxidiophora sp. 03 18S rRNA 1064 AY212811.1 gPyxidiophora arvernensis18S rRNA +type I intron424 U43339.1 (h)[(a) MAHDI & DONACHIE, 2006 – unpublished; b WEIR & HUGHES, 2002; c WEIR & BLACKWELL, 2001b; d WEIR & BLACKWELL,2001a; (e) WEIR & BLACKWELL, 2000 – unpublished; (f) SCHOCH, 2008 – unpublished; g HENK, WEIR & BLACKWELL, 2003; (h)Jones & Blackwell, 1995 – unpublished.]P a g e | 33
2. MATERIALS & METHODS2.1. FUNGUS, HOST AND ORIGINCarabid hosts of Laboulbenia species were collected in November 2008 and2009 (July – December) by means of hand capturing. All collecting sites aresituated in Hingene (Belgium, Prov. Antwerpen, N51°06‟ – E4°12‟), atSchellandpolderdijk along the river Schelde.The collecting sites can be categorized as alluvial areas (polders). These areasare not flooded daily, being protected by dykes; they have fine alluvial soilsthat can become relatively dry in summer. The sampled alluvial areas consistmostly of poplar plantations or mixed forests with mainly Alnus, Fraxinus,Quercus and Acer.2.2. MORPHOLOGICAL PROTOCOL AND DNA EXTRACTION2.2.1. INTRODUCTIONThe hosts were narcotized in a pot filled with chloroform gas (insects had nocontact with liquid chloroform). Screening of the hosts was done by using bothbinocular and stereomicroscope (50x). Separation of infected and uninfectedhosts had to be done with precision. Carabid hosts were identified to specieslevel, using the identification key of BOEKEN (1987).From this point, different strategies were tried out, in order to improve themethodology. The protocols will be discussed below, in chronological order.2.2.2. PROTOCOL I (BASED ON WEIR & BLACKWELL, 2001b)Thalli were aseptically removed from each host using a sterile micropin andtransferred to a 2 µL drop of Milli-Q H2O on a sterile microscope slide. An 18 x18 mm coverslip was placed over the thallus. Hosts were observed using aNikon SMZ800 stereoscopic zoom microscope (binocular); a Nikon Eclipse E600research microscope was used for visual inspection of the thalli. Thalli wereidentified to species level using MAJEWSKI (1994), and photographed with NikonDigital Camera DXM1200. After photographing, the thalli were crushedbetween the two slides. A razor blade was used to remove the coverslip.Visual inspection was needed in order to find the thalli on the coverslip or themicroscopic slide. The crushed material was hydrated in 20 μL of extractsolution (cfr. Figure VIII for composition).99 µL Milli-Q H2O + 1 µL Triton (100%) 100 µL 1% Triton detergent1 µL 1% Triton + 19 µL Milli-Q H2O 20 µL extract solutionFigure VIII: Composition of extract solution.P a g e | 34The thalli were transferred into 2mL tubes using a sterile micropin. Glass beads(0,25-0,50 mm in diameter) were added to the tube. The content of the tubewas additionally crushed in a bead beater (3 x 90 sec; 30x/s) (RETSCH Mixer MillMM200, Haan, Germany). Centrifugation took place for 2 minutes at 14.000rcf. 30 µL Milli-Q H2O was added to the tube, whereupon the tube was placed
Page 3: LABOULBENIALESEXPLORING AND TESTING
Page 7 and 8: PART IVPRELIMINARY CHECKLIST OF LAB
Page 9 and 10: SAMENVATTINGINLEIDINGLaboulbeniales
Page 11 and 12: PART IGENERAL INTRODUCTIONTHESIS OU
Page 13 and 14: fifth volume, therefore the sixth v
Page 16 and 17: 1.3.3. THE PERITHECI UMAscospores o
Page 18 and 19: The identity of appendages, togethe
Page 20 and 21: Figure IV: Position of Laboulbeniom
Page 22 and 23: LUMBSCH & HUHNDORF (2007) distingui
Page 25 and 26: Substrate is the intermediate facto
Page 27: CAVARA (1899, ref. in BENJAMIN, 197
Page 35: 1. INTRODUCTION1.1. DIFFICULTIES FO
Page 39 and 40: 2.2.6. PROTOCOL V: DIRECT PCROne th
Page 41 and 42: P a g e | 38chance. Thus, the lower
Page 43 and 44: coll1133 Laboulbenia collae IV 608l
Page 45 and 46: Cladosporium (Ascomycota, Dothideom
Page 47 and 48: 5. CONCLUSION AND SUGGESTIONSMany a
Page 49 and 50: 1. INTRODUCTION1.1. FORENSIC ENTOMO
Page 51 and 52: Different groups of carrion beetles
Page 53 and 54: The 1.319 ha Meerdaal Forest has be
Page 55 and 56: 2.2. MEERDAAL FOREST(DA TA NA TI O
Page 57 and 58: approximately 2.700 are found in Eu
Page 59 and 60: Order ColeopteraSuborder PolyphagaI
Page 61 and 62: Dermestidae 8 2 23 (3) / 22 (4)Elat
Page 63 and 64: Table XXIV: Family Cerambycidae fro
Page 65 and 66: Table XXVIII: Family Dermestidae fr
Page 67 and 68: Adults prey on other flower-visitin
Page 69 and 70: Table XXXV: Family Scarabaeidae fro
Page 71 and 72: Creophilus Samouellemaxillosus (Lin
Page 73 and 74: 4.3. LABOULBENIALES OF CARRION BEET
Page 75 and 76: The knowledge about the preferences
Page 77 and 78: Table XLII: Comparison of carrion b
Page 79 and 80: 5. CONCLUSION AND SUGGESTIONSNo spe
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Page 83 and 84: In the 40s, Middelhoek studied and
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Screening of the insects was done w
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3. RESULTS3.1. PARASITE-HOST LISTA
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Figure XV: Detail of thallus of Lab
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isodiametric or slightly elongate,
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occurs in the lower receptacle (cel
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Discussion:Two species of the genus
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anches (15-)34-38(-40) µm long, te
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5. CONCLUSION AND SUGGESTIONSIn The
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P a g e | 100ADRIAENS, T. & GYSELS,
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transmission, habitat preference an
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STALPERS, J.A., VILGALYS, R., AIME,
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MELIS, C., TEURLINGS, I. LINNELL, J
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SCHILTHUIZEN, M. & VALLENDUUK, H. (
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