真菌-酵母细胞内氧化应激活性氧高质荧光测定试剂盒文献

文档简介AbstractBackground:PolyporusumbellatussclerotiahavebeenusedasadiureticagentinChinaforovertwothousandyears.AshortageofthenaturalP.umbellatushaspromptedresearcherstoinducesclerotialformationinthelaboratory.Methodology/PrincipalFinding:P.umbellatuscultivationinasawdust-basedsubstratewasinvestigatedtoevaluatetheeffectoflowtemperatureconditionsonsclerotialformation.Aphenol-sulfuricacidmethodwasemployedtodeterminethepolysaccharidecontentofwildP.umbellatussclerotiaandmyceliaandsclerotiagrowninlow-temperaturetreatments.Inaddition,reactiveoxygenspecies(ROS)content,expressedasthefluorescenceintensityofmyceliaduringsclerotialdifferentiationwasdetermined.AnalysisofROSgenerationandsclerotialformationinmyceliaaftertreatmentwiththeantioxidantssuchasdiphenyleneiodoniumchloride(DPI),apocynin(Apo),orvitaminCwerestudied.Furthermore,macroscopicandmicroscopiccharacteristicsofsclerotialdifferentiationwereobserved.Sclerotiawerenotinducedbycontinuouscultivationat25uC.Thepolysaccharidecontentoftheartificialsclerotiais78%ofthatofwildsclerotia.Inthelow-temperaturetreatmentgroup,thefluorescentintensityofROSwashigherthanthatoftheroomtemperature(25uC)groupwhichdidnotinducesclerotialformationallthroughthecultivation.TheantioxidantsDPIandAporeducedROSlevelsanddidnotinducesclerotialformation.Althoughtheconcentration-dependenteffectsofvitaminC(515mgmL21)alsoreducedROSgenerationandinhibitedsclerotialformation,usingalowconcentrationofvitaminC(1mgmL21)successfullyinducedsclerotialdifferentiationandincreasedROSproduction.Conclusions/Significance:ExposuretolowtemperaturesinducedP.umbellatussclerotialmorphogenesisduringcultivation.LowtemperaturetreatmentenhancedROSinmycelia,whichmaybeimportantintriggeringsclerotialdifferentiationinP.umbellatus.Moreover,theapplicationofantioxidantsimpairedROSgenerationandinhibitedsclerotialformation.OurfindingsmayhelptoprovidenewinsightsintothebiologicalmechanismsunderlyingsclerotialmorphogenesisinP.umbellatus.Citation:XingY-M,ZhangL-C,LiangH-Q,LvJ,SongC,etal.(2013)SclerotialFormationofPolyporusumbellatusbyLowTemperatureTreatmentunderArtificialConditions.PLoSONE8(2):e56190.doi:10.1371/journal.pone.0056190Editor:PratibhaV.Nerurkar,CollegeofTropicalAgricultureandHumanResources,UniversityofHawaii,UnitedStatesofAmericaReceivedAugust16,2012;AcceptedJanuary7,2013;PublishedFebruary20,2013Copyright:2013Xingetal.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited.Funding:TheresearchwasfinanciallysupportedbytheNationalNaturalSciencesFoundationofChina(No.30830117,31201666),theInternationalScienceandTechnologyCooperationProjectsofChina(No.2011DFA31260)andtheNationalResearchFoundation(NRF20110016999).Thefundershadnoroleinstudydesign,datacollectionandanalysis,decisiontopublish,orpreparationofthemanuscript.CompetingInterests:Theauthorshavedeclaredthatnocompetinginterestsexist.*E-mail:sxguo2006@yahoo.com.cn(SXG);cl王@implad.ac.cn(CLW)