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1、JournalofMicrobiologicalMethods81(2010)6–10ContentslistsavailableatScienceDirectJournalofMicrobiologicalMethodsjournalhomepage:www.elsevier.com/locate/jmicmethOptimizedmicroplateβ-1,3-glucanaseassaysystemforTrichodermaspp.screeningMarceloHenriqueSollerRamada,
2、FabyanoÁlvaresCardosoLopes,CiranoJoséUlhoa⁎,RobertodoNascimentoSilvaLaboratóriodeEnzimologia,DepartamentodeBioquímicaeBiologiaMolecular,UniversidadeFederaldeGoiás(ICB),Goiânia,GO,CEP74001-970,BrazilarticleinfoabstractArticlehistory:β-1,3-glucanaseisanimportan
3、tcellwall-degradingenzymeinvolvedinmycoparasitismbyTrichodermaReceived8November2009spp.duringantagonismagainstphytopathogenicfungi.Asimplemicroplate-basedmethodtoassayβ-1,3-Receivedinrevisedform8January2010glucanaseactivityisdescribedhereasanalternativetoanex
4、pensivetube-assaymethod.ThereactionAccepted9January2010volumeofthemicro-assaywasreducedto130µlfromthe1150µlusedinthestandardβ-1,3-glucanaseAvailableonline21January2010macro-assay.Statisticalanalysesshowedsignificantdifferenceinsensitivitybetweenthemicro-andthe
5、macro-assay.Themicro-methodwasoptimizedusingtheResponseSurfaceQuadraticModel.ThesensitivityKeywords:Biocontroloftheoptimizedmicro-methodwasshowntobefour-foldgreaterthanthemacro-assayandtwo-foldLaminarinhigherthanthemicro-assay.Theoptimizedmicro-assaywassignifi
6、cantlymoresensitiveinalloftheMicroplateassaytwentyexaminedisolatesduringTrichodermaspp.β-1,3-glucanasescreening.WeconcludethatthisTrichodermamodifiedandoptimizedmethodismoreconvenient,faster,cheaperandmorereproduciblethantheβ-1,3-glucanasetraditionaltube-assay
7、.©2010ElsevierB.V.Allrightsreserved.1.Introductionandcanthereforebeusedasbiocontrolagents(Howell,2003).ThemechanismofTrichodermaspp.actioncaninvolveantibiosis,β-1,3glucanasehasbeenreportedinagreatvarietyoforganisms,nutrient/spacecompetitionandmycoparasitism,o
8、neofthemajorincludingarchaea(Gueguenetal.,1997),bacteria(Watanabeetal.,antagonisticmechanismsdisplayedbyTrichoderma(Kubiceketal.,1992),actinomycetes(Prapagdeeetal.,2008),yeast(Xuetal.,200
