Penicillium 2012 - CBS - KNAW

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Penicillium 2012 - CBS - KNAW

10/4/2012Exudate production in PenicilliumSoluble pigment production in PenicilliumSummery of presentation I• Macro-morphology:Colony colour (obverse/reverse)Colony sizeExudateSoluble pigment• Micro-morphology:Branching patternShape phialide• Ehrlich test3


10/4/2012Flask-shapedPaecilomycesLanceolate = aceroseconidiaphialidesconidiaStipePenicillium conidiophore = Penicillus4


10/4/2012BRANCHING PATTERNS IN PENICILLIUMmonoverticillatebiverticillatebiverticillateterverticillateDifferences in branching pattern and shape phialides!Penicillium glabrumMonoverticillate(= unbranched, simple)Penicillium citrinumBiverticillate(=one-stage branched)5


10/4/2012Penicillium corylophilumBiverticillate(=one-stage branched)Penicillium brevicompactumTerverticillate(=Two-staged branched)Penicillium expansumTerverticillate(=Two-staged branched)6


10/4/2012Flask-shapedPaecilomycesLanceolate = acerosePaecilomyces• Phialides with broadbase and long narrowneck• Conidiophorebranching irregularly• Cultures mostly notgreen• ThermophilicPaecilomyces variotii7


10/4/2012PenicilliumBiverticillium• Penicillus biverticillate= one-staged branched• Phialides acerose =lanceolateFlask-shaped phialidesLanceolate phialides8


10/4/2012Colony surfaces in PenicilliumCultivation for identification• Czapek Yeast agar (CYA):Macro-morphology• Czapek Yeast agar (CYA) incubated at 30°CColony diameter (and ratio with 25°C)• Malt Extract Agar (MEA): macro- andmicromorphology• Creatine agar (CREA), used for identification ofterverticillate Penicillia• Yeast Extract Agar (YES), only macro-morphology• Incubation 7 days at 25°C9


10/4/2012CREA: Creatine Sucrose agarCreatine(1 H 2O)SucroseKClMgSO 4.7H 2OFeSO 4.7H 2OK 2HPO 4.3H 2OBromocresol purpleAgarDistilled water3 g30 g0.5 g0.5 g0.01 g1.3 g0.05 g15 g1000 mlFinal pH 8.0 0.2 (adjust after medium is autoclaved).NOTE: a modification of CREA (=CRE) with 1.6 gK 3PO 4.7H 2O can also be used.Examples growth on CREAAcid / Base production on CREA7 days12 days 12 days12 days12 days12 days10


10/4/2012Ehrlich reaction• Filter paper test; detection of presence ofindole metabolites• Plug for 7 days old CYA culture• Place filterpaper with Ehrlich reagens onthe mycelium side of the plug (propeller)• Read after 5 and 10 minutes• No reaction / violet (/yellow / brown)Ehrlich reaction patternsViolet reactionsNo reactionSummary of presentation II• Macro-morphology:Colony color (obverse/reverse)Colony sizeExudateSoluble pigment• Micro-morphology:Branching patternShape phialide• Ehrlich test11


10/4/2012CYA, obverse - 30°CMacroscopical featuresMEA obverseCYA obverseCYA reverseYES obverseYES reverseCREA obverseMicroscopic preparation of Aspergillus and PenicilliumLactic acid with analine blueDrop of alcoholPreparations from Malt Extract AgarSIM 49: Penicillium subgenus Penicillium:new taxonomic schemes, mycotoxins andother extrolites (accessible via CBShomepage; “Studies In Mycology”)On-line database about Penicillium subg. Penicillium:http://www.cbs.knaw.nl/databases/index.htm (select Penicillium database)12

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