1、AJS(AcidifiedJM:SE)FreshwateralgaDunaliellaacidophilaMediumAcidified97:3mixtureofJMandSE2Seeseparaterecipes(JMandSE2).Forapproximately1litreoffinalmedium,mix970mlJMwith30mlSE2.Add10mlofconcH2SO4togiveapHofapproximately1.5.Autoclaveat15psifor15minutes.ANT(AntiasMedium)MarinealgaeStockTracemetalsstock

2、solution(chelated)perlitreEDTANa2.2H2O3.24gFeCl3.4H2O1.08gMnSO4.4H2O0,450gZnSO4.7H2O0,230gNa2MoO4.2H2O0,097gCuSO4.5H2O0.01gCoSO4.7H2O0,0056gMakeupto1litrewithdistilledwaterandadjustpHto7.6-7.8withdiluteHClorNaOH.Storefrozen.MediumperlitreKNO30.05gNaH2PO4.2H2O0,0078gTristris(hydroxymethyl)aminomethan

3、e1,0gGlycine0.3gTracemetalsstocksolution(chelated)2.5mlThiamineHCl500.0聞Cyanocobalamin2.0聞Biotin1.0聞Filterednaturalseawater800.0mlMakeupto1litrewithdistilledwaterandautoclaveat15psi.FinalpHshouldbe7.6-7.8.RefAntia,Cheng&Taylor(1969)ASW(ArtificialSeawater)MarinealgaeNaNO3Na2HPO4K2HPO4(2)Vitaminso

4、lutionperlitreBiotin0.0002gCalciumpantothenate0.02gCyanocobalamin0.004gFolicacid0.0004gStocks(1)Extrasaltsperlitre30.0g1.2g1.0gInositol1.0gNicotinicacid0.02gThiamineHCl0.1gThymine0.6gMaybestoredfrozenat-20CMediumperlitre*UltramarineSyntheticaseasalts33.6gExtrasaltsstocksolution(1)3.75mlVitaminstocks

5、olution(2)2.5mlSoilextract(SE1-seerecipe)25.0mlTricine0.50gMakeupto1litrewithdistilledwaterandadjustpHto7.6-7.8with1MNaOHorHCl.Autoclaveat15psi.Supply*WaterlifeResearchIndustriesLtd,476BathRoad,Longford,WestDrayton,MiddlesexUB70ED,UK.Tel(01753)685696Fax(01753)685437ASW+barley(ArtificialSeawater+barl

6、eygrains)MarinedinoflagellateOxyrrhisMediumFollowtherecipeforASWbutaddapproximatelyonegrainofbarleytoeach25mlofpreparedmediumpriortoautoclaving.2ASW(doublestrengthArtificialSeawater)MarinealgaeMediumFollowtherecipeforASWbutadd35gNaClperlitrepriortoautoclaving.ASW:BGMarinealgaeMedium1:1mixtureSeesepa

7、raterecipes.Mixaseptically.ASW:SESMarinealgaeMedium3:1mixtureSeeseparaterecipes.Mixbeforeautoclaving.ASWP(ArtificialSeawaterforProtozoa)MarineprotozoaStockSolution(1)NaNO3perlitre5.625gNa2HPO4K2HPO40.225g0.188gMediumperlitre*UltramarineSyntheticaseasaltsTricine0.533.6gStocksolution(1)Soilextract(SE2

8、-seerecipe)10.0ml50.0mlMakeupto1litrewithdeionizedwaterandadjustpHto7.6-7.8with1MNaOHorHCl.Autoclaveat15psifor15minutes.Supply*asinASWrecipeBB(BoldsBasalMedium)Stocksper200ml(1)NaNO35.0gMgSO4.7H2O1.5g(3)NaCl0.5g(4)K2HPO41.5gKH2PO43.5g(6)CaCl2.2H2O0.5g(7)Traceelementssolution:perlitreZnSO4.7H2O8.82gM

9、nCl2.4H2O1.44gMoO30.71gCuSO4.5H2O1.57gCo(NO3)2.6H2O0.49gMayneedautoclavingtodissolve.per100ml(8)H3BO31.14g(9)EDTA-KOHsolution:EDTANa25.0gKOH3.1gFreshwateralgaeperlitre4.98g1.0mlMediumStocksolutions1-610.0mleachStocksolutions7-101.0mleachMakeupto1litrewithdeionizedwater.Foragar,add15.0gperlitreof*Bac

10、teriologicalAgar(OxoidL11).Autoclaveat15psifor15minutes.Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKBB:MErdsFreshwater/brackishwateralgaeMedium8:2mixtureor1:1mixtureSeeseparaterecipes(BBandMErds).Mixthenautoclaveat15psifor15minutes.BG(Blue-GreenMedium)MarinecyanobacteriaStocks(1)Extranutri

11、entsaltsper100mlNaNO33.00gNa2HPO40.12gK2HPO40.10g(2)Tracemetalsolutionper100mlH3BO30.286gMnCl2.4H2O0,022gNa2MoO4.2H2O0,039gCuSO4.5H2O0,008gCo(NO3)2,6H2O0,005gMediumThismediumismadeupin2parts:PART 1perlitreTricine0.50gSoilextract(SE1-seerecipe)25.00mlExtranutrientsalts(1)3.75mlMakeupto1litrewithfilte

12、rednaturalseawaterandadjustpHto7.6-7.8with1MNaOHorHCl.PART 2perlitreNaNO31.500gK2HPO4.3H2O0,040gMgSO4.7H2O0,075gCaCl2.2H2O0,036gCitricacid0.006gAmmoniumferriccitrategreen(10)FeSO4,7H2OconcH2SO4EDTANa20.001gNa2CO30.020gTracemetalsolution(2)1.00mlMakeupto1litrewithdistilledwaterandadjustpHto7.4.FinalA

13、utoclaveParts1and2separatelyat15psi,allowtocoolthenmixaseptically.Foragarplates,add15gnon-nutrientagarperlitre.BG11(Blue-GreenMedium)FreshwateralgaeandprotozoaStocksperlitre(1) NaNO315.0gper500ml(2) K2HPO42.0g(3) MgSO4.7H2O3.75g(4) CaCl2.2H2O1.80g(5) Citricacid0.30g(6)Ammoniumferriccitrategreen(枸檬酸鐵

14、鏤，檸檬酸鐵鏤)0.30g(7) EDTANa20.05g(8) Na2CO31.00g(9) TracemetalsolutionperliterH3BO32.86gMnCl2.4H2O1.81gZnSO4.7H2O0.22gNa2MoO4.2H2O0.39gCuSO4.5H2O0.08gCo(NO3)2.6H2O0.05gMediumStocksolution1100.0mlStocksolutions2-810.0mleachStocksolution91.0mlMakeupto1litrewithdeionizedwater.AdjustpHto7.1with1MNaOHorHCl.F

15、oragaradd15.0gperlitreof*BacteriologicalAgar(OxoidL11).Autoclaveat15psifor15minutes.Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKRefStanieretal,(1971)CH(ChalkleysMedium)FreshwaterprotozoaStocks(1)NaClper100ml2.0g(3)CaCl20.12gMediumStocksolutions1-35.0mleachMakeupto1litrewithdeionizedwater.A

16、utoclaveat15psifor15minutes.CHM(ChilomonasMedium)FreshwaterprotozoaMediumperlitreSodiumacetatetrihydrate1.0g*Lab-Lemcopowder(OxoidL29)1.0gAddtheaboveconstituentsto1litreofdeionizedwater.Autoclaveat15psifor15minutes.Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKCMA(CornMealGlucoseAgar)Freshwa

17、terprotozoaMediumperlitre*Cornmealagar(OxoidCM103)17.0gD-glucose2.0g*Yeastextract(OxoidL21)1.0gDispersetheagarincolddeionizedwater.Applyheat,stirringconstantly.Duringheatingaddglucoseandyeastextract.Bringcontentstoboil.Transfermoltenagartosuitablevessels.Sterilizebypressurecookingat10psifor15minutes

18、.Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKDM(DiatomMedium)FreshwateralgaeStocksper200ml(1)Ca(NO3)2.4H2O4.00g(2)KH2PO42.48g(3)MgSO4.7H2O5.00g(4)NaHCO33.18g(5)EDTAFeNa0.45gEDTANa20.45g(6)H3BO30.496gMnCl2.4H2O0,278g(NH4)6Mo7O24.4H2O0.20gCyanocobalamin0.008gThiamineHCl0.008gBiotin0.008g*(8)

19、NaSiO3.9H20(SigmaS4392)11.4gMakeupto500mlwithdistilledwater.PART2MediumStocksolutions1-81.0mleachMakeupto1litrewithdeionizedwater.AdjusttopH6.9with1MHCl.Dispensetosuitablevesselsandautoclaveat15psifor15minutes.Supply*SigmaChemicalCo,FancyRoad,Poole,DorsetBH177TG,UKRefBeakes,Canter&Jaworski(l988)

20、EG(EuglenaGracilisMedium)FreshwateralgaeandprotozoaStock(1)CaCl2stocksolution:CaCl2perlitreMedium1.0gperlitreSodiumacetatetrihydrate*Lab-Lemcopowder(OxoidL29)1.0g1.0g*Tryptone(OxoidL42)*Yeastextract(OxoidL21)CaCl2stocksolution(1)2.0g2.0g10.0mlAddconstituentsaboveandmakeupto1litrewithdeionizedwater.F

21、oragaradd15gperlitre*BacteriologicalAgar(OxoidL11).Autoclaveat15psifor15minutes.Supply*UnipathLtd,WadeRoad,Basingstoke,HantsRG240PW,UKEG:JMMedium1:1mixtureSeeseparaterecipes(EGandJM).Mixthenautoclaveat15psifor15minutes.E27(E27Medium)Marinealgae,includingsterilitytestingMediumThismediumismadeupin2partstoavoidprecipitation:PART1Soilextract(perlitreSE1-seerecipe)25.0mlKNO30.050gK2HPO4MgSO4.7H200.005g0.00