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Chapter2
CellBiology第一页,共九十二页。
2.1Overviewofthestructureofmicrobialcells2.2Procaryoticcellwall2.3Cytoplasmicmembrane2.4Cellulargeneticinformation2.5Cytoplasmicmatrix–RibosomeandInclusions2.6Componentsexternaltothecellwall2.7Bacterialendospores2.8ComparisonoftheprokaryoticandeukaryoticcellChapterOutline第二页,共九十二页。ConceptsProkaryotesaresmallandsimpleinstructurewhencomparedwitheukaryotes,yettheyoftenhavecharacteristicshapeandsize.
Prokaryoticgeneticmaterialislocatedinanareacalledthenucleoidandisnotenclosedbyamembrane.Theprokaryoticcellwallalmostalwayshaspeptidoglycanandischemicallyandmorphologicallycomplex.第三页,共九十二页。AprocaryoticcellAeucaryoticcell2.1OverviewofCellStructure第四页,共九十二页。3.TheircellwallalmostalwayscontainthecomplexpolysaccharidepeptidoglycanTheProkaryoticCell1.Theirgeneticmaterial(DNA)isnotenclosedwithinamembraneandtheylackothermembrane–boundedorganelles2.TheirDNAisnotassociatedwithhistidine4.Theyareverysmall!!第五页,共九十二页。Schematicdiagramofabacterialcell第六页,共九十二页。Size:
Mostbacteriafallwithinarangefrom0.2to2.0umindiameterandfrom2to8uminlengthArod-shapedprokaryoteistypicallyabout1-5micrometers(μm)longandabout1μmwideMicroorganismsingeneralareverysmallandarecompletelyinvisibletothenakedeye.
Acyanobacterium8x50um第七页,共九十二页。sizecomparisonofmicroorganismsVisibility
scaleMeters
RelativesizeofMicrobesProkaryotesEukaryotesVirusesNakedeyeLightmicroscopeElectronmicroscope第八页,共九十二页。acellincreasesinsize,itssurfacearea–to–volumeratiodecreasesSurfaceareaandvolumerelationshipsincells第九页,共九十二页。spirallumShape:
Bacteriahaveafewbasicshapesspherical
coccusRod-shaped
bacillus第十页,共九十二页。2.2ProkaryoticCellWallCellWall第十一页,共九十二页。Thecellwallofthebacterialcellisacomplex,semi-rigidstructurethatisresponsibleforthecharacteristicshapeofthecell.Thecellwallsurroundstheunderlying,fragileplasma(cytoplasmic)membraneandprotectsitandinternalpartsofthecellfromadversechangesinthesurroundingenvironment.Almostallprokaryoteshavecellwalls.第十二页,共九十二页。Gram+Gram-SchematicdiagramofbacterialcellwallsBacteriacanbedividedintotwomajorgroups,calledgram-positiveandgram-negative.Theoriginaldistinctionbetweengram-positiveandgram-negativewasbasedonaspecialstainingprocedure,theGramstain第十三页,共九十二页。TheGram-positivecellwallhasapeptidoglycanlayerthatisrelativelythick(ca.40nm)andcomprisesapproximately90%ofthecellwall.ThecellwallsofmostGram-positiveeubacteriaalsohaveteichoicacids.Gram-positiveCellWall第十四页,共九十二页。StructureoftheRepeatingUnitinPeptidoglycanTheseconstituentsareconnectedtoformarepeatingstructure,theglycantetrapeptide.Peptidoglycaniscomposedoftwosugarderivatives,N-acetylglucosamine(NAG)andN-acetylmuramicacid(NAM),andasmallgroupofaminoacidsconsistingofL-alanine,D-alanine,D-glutamicacid,andeitherlysineordiaminopimelicacid(DAP).第十五页,共九十二页。第十六页,共九十二页。Peptideandglycanunitsareconnectedinformationofthepeptidoglycansheet第十七页,共九十二页。Gram-positiveBacteriafrequentlyhaveacidicpolysaccharidescalledteichoicacidsattachedtotheircellwall.Thetermteichoicacidsincludesallwall,membrane,orcapsularpolymerscontainingglycerophosphateorribitolphosphateresidues.ThesepolyalcoholsareconnectedbyphosphateestersandusuallyhaveothersugarsandD-alanineattached.
TeichoicAcids第十八页,共九十二页。Teichoicacidsandlipoteichoicacidsarearrangedintheoverallwallstructureofgram-positiveBacteria.TeichoicacidLipoteichoicacid第十九页,共九十二页。TheGram-negativecellwallisathinlayerattachedtoanoutermembranevialipoproteins.Theoutermembranecontains
phospholipidonitsinnersurfaceand
lipopolysaccharide(LPS)
on
itsoutersurface.Thespacebetweentheoutermembraneandthe
cytoplasmicmembraneiscalledthe
periplasmicspace.Teichoic
acidsdonotoccurinGram-negativebacterialcellwalls..
第二十页,共九十二页。第二十一页,共九十二页。OsidechainCorepolysaccharideLipidAChemicalStructureofLipopolysaccharide第二十二页,共九十二页。MolecularModelofE.coliLipopolysaccharide第二十三页,共九十二页。Thebondsbetweenthecarbohydratesinpseudopeptidoglycanare
β1-3insteadof'β1-4
asinpeptidoglycan.
CellWallsofArchaebacteriaThearchaebacteria
donotcontainpeptidoglycan
intheircellwallsasoccursineubacteria.N-acetylmuramicacidandD-aminoacids
arenotfound
inthecellwallsofarchaebacteria.(Differencesfromeubacteria)第二十四页,共九十二页。Somearchaebacteriahavewallscomposedof
pseudopeptidoglycan,
whichresemblesthepeptidoglycanofeubacteriabut
containsN-acetyltalosaminuronicacid
insteadofN-acetylmuramicacidand
L.-aminoacids
insteadoftheD-aminoacidsineubacterialcellwalls.第二十五页,共九十二页。ProtoplastFormation
Peptidoglycancanbedestroyedbycertainagentsforinstancelysozyme,thatbreaksthe1,4-glycosidicbondsbetweenN-acetylglucosamineandN-acetylmuramicacidinthemolecule.第二十六页,共九十二页。Thedifferencebetweengram-positiveandgram-negativebacteriaisduetothephysicalnatureoftheircellwalls.Ifthecellwallisremovedfromgram-positivebacteria,theybecomegramnegative.
第二十七页,共九十二页。Thepeptidoglycanseemstoactasapermeabilitybarrierpreventinglossofcrystalviolet.Gram-negativepeptidoglycanisverythin,notashighlycross-linked,andhaslargerpores.Alcoholtreatmentalsomayextractenoughlipidfromthegramnegativewalltofurtherincreaseitsporosity.Forthesereasons,alcoholmorereadilyremovesthepurplecrystalviolet-iodinecomplexfromgram-negativebacteria.TheMechanismofGramStaining第二十八页,共九十二页。
ProceduresofGramStaining第二十九页,共九十二页。GrampositiveorGramnegative?第三十页,共九十二页。第三十一页,共九十二页。Structureofcytoplasmicmembrane2.3CytoplasmicMembraneFunctionofcytoplasmicmembraneMembrane第三十二页,共九十二页。B.
Colorizedelectronmicrographof'thecytoplasmicmembrane(CM)ofthebacteriumBacillussubtilis
revealsthecharacteristicrailroadtrackappearanceofthislipidbilayer.
StructureofCytoplasmicMembraneItisatypicalUNITMEMBRANE!A.
Thetypicalcytoplasmicmembraneofprokaryoticandeukaryoticcellsisalipidbilayer,asillustratedhereshowingtheorientationsofthehydrophilic(tanspheres)andhydrophobic(black)endsofphospholipidsthatmakeupthisstructure.第三十三页,共九十二页。Thecytoplasmicmembrane,ahighlyselectivebarrier,isconstructedprincipallyoflipid,withinwhichcertainproteinsareembedded.Membranescontainboth
lipidsandproteins,althoughtheexactproportionsoflipidandproteinvarywidely.第三十四页,共九十二页。Diagramofthestructureofcytoplasmicmembrane第三十五页,共九十二页。3.Energyconservation-siteofgenerationanduseoftheprotonmotiveforce.
FunctionofMembrane1.Permeabilitybarrier
-preventsleakageandfunctionasgatewayfortransportofnutrientsintoandoutofthecell.2.Proteinanchor-siteofmanyproteinsinvolvedintransport,bioenergetics,andchemotaxis.第三十六页,共九十二页。
Rateofpermeability*
100
0.1
0.001
0.001
0.000001
0.0000001
0.00000001
Substance
Water
Glycerol
Tryptophan
Glucose
Chlorideion(Cl-)
Patassiumion(K+)
Sodiumion(Na+)
*Relativerate:Permeabilitywithrespecttopermeabilityofwatergivenas100
Comparativepermeabilityofmembranemoleculestovarious
第三十七页,共九十二页。IntracellularMembraneSystemBacteriacellsdon’tcontainmembrane-enclosedorganelles.However,bacteriamayhavespecializedinvaginationsofthecytoplasmicmembrane.Theirfunctionmaybetoprovidealargermembranesurfaceforgreatermetabolicactivity.第三十八页,共九十二页。StructureofMesosomeMesosomemaybeinvolvedin
wallformation
duringdivisionorplayarolein
chromosomereplication
anddistributiontodaughtercells.Itmayalsobeinvolvedin
secretoryprocesses第三十九页,共九十二页。2.4CellularGeneticInformationNucleoid第四十页,共九十二页。BacterialChromosomeSupercoilingandchromosomestructureChromosomalcopynumber2.Plasmids第四十一页,共九十二页。Micrographofabacteriumshowingthenucleoidregion(green)withinthecytoplasmwherethebacterialchromosomeoccurs第四十二页,共九十二页。ThebacterialchromosomeisacircularDNAmacromoleculeexcept
inStreptomyces
whereitislinearand
Rhodobacter
sphaffoides,whichhastwoseparatechromosomes.
BacterialChromosomeThebacterialchromosomeisusuallya
singlecovalentlyclosedcircular
molecule.Theterm
nucleoidisusedtodescribeaggregatedDNAintheprokaryoticcell.第四十三页,共九十二页。Rangeofgenomesizesinvirousgroupsoforganismsandtheorganellesofeukarya第四十四页,共九十二页。Thebacterialchromosomeandsupercoiling:第四十五页,共九十二页。ExampleofE.colicellThereareover
50supercoileddomain
intheE.colichromosome.ThetotalamountofDNAisabout
4600kb.IfthetotalDNAisopenedandlinearized,itwouldbe
1mminlength.Thethecellisonly
about2-3umlong.
SotopackagethismuchDNAintothecellrequiresthat
theDNAbehighlysupercoiled.第四十六页,共九十二页。ElectronmicrographofanisolatednucleoidreleasedfromE.coli.
第四十七页,共九十二页。Chromosomecopynumber
Bacteriathatreproduceasexuallyaretypically
haploid
ingeneticcomplement.Rapidlygrowingcellscontain
morethan
1copy
ofthechromosome,andonlywhencellgrowthhasceaseddoesthechromosomenumberapproachonepercell.第四十八页,共九十二页。
Reproductionofabacterialcellrequiresthereplicationofthebacterialchromosome.ThemicrographshowsthesequenceofsynthesisofnewcircularloopsofdoublehelicalDNA.Bacterianormallyreproducebybinaryfission.Theinwardgrowthoftheseptumdividestheparentcelltoproducetwoequal-sizedprogenycells.第四十九页,共九十二页。Plasmidsdon’tcontainthegeneticinformationfortheessentialmetabolicactivitiesofthecell,buttheygenerallydocontaingeneticinformationforspecialfeatures.
PlasmidProkaryoticcellshavesmallextra-chromosomal
geneticelementscalled
plasmids.第五十页,共九十二页。ResistantplasmidsColplasmidsConjugativeplasmidsMetabolicplasmids
MajorTypesofPlasmids第五十一页,共九十二页。2.5CytoplasmicMatrixRibosomeandInclusionsRibosomeCarbonstoragepolymers–PHBandglycogenPhosphatepolymersSulfurGranulesGasVacuoles第五十二页,共九十二页。Alleucaryoticandprocaryoticcellscontainribosomes,whichfunctionasthesitesofproteinsynthesis.Ribosomesarecomposedoftwosubunits
Procaryoticribosomesarecalled70Sribosomes,andthoseofeucaryoticcellsareknownas80Sribosomes
RibosomesTheletterSreferstoSvedbergunits,whichindicatetherelativerateofsedimentationduringultra-high-speedcentrifugation第五十三页,共九十二页。第五十四页,共九十二页。
Withinthecytoplasmofprocaryotic(andeucaryotic)cellsareseveralkindsofreservedeposits,knownasinclusions.Someinclusionsarecommontoawidevarietyofbacteria,whereasothersarelimitedtoasmallnumberofspeciesandthereforeserveasabasisforidentification.Amongthemoreprominentbacterialinclusionsarethefollowing:InclusionCarbonstoragepolymers–PHBandglycogenPhosphatepolymersSulfurGranulesGasVacuoles第五十五页,共九十二页。Polyhydroxybutyricacid(PHB)PHBisa
lipidlikecompound
-oneofthemostcommoninclusionbodiesinprokaryoticorganisms.PHBiscommonlyfoundasastoragematerialand
uniquetobacteria
Glycogenisa
starchlikepolymer
ofglucosesubunits.GlycogengranulesareusuallysmallerthanPHBgranules.AVibriospecies第五十六页,共九十二页。Manymicroorganismsaccumulategranulesofpolyphosphate,whicharelargereservesof
inorganicphosphates
thatcanbeusedinthesynthesisofATPPolyphosphategranule
inabacterialcellAPseudomonasspecies第五十七页,共九十二页。ThesulfurglobulesinsidethecellsofpurplesulfurbacteriumChromatiumbuderiSomebacteria,includingmanyphotosyntheticbacteria,accumulate
elementalsulfurgranules
asaresultoftheirmetabolism.第五十八页,共九十二页。Gasvacuoles(blue)andstoragegranules(red)inthecyanobacteriumMicrocystisTheformationofgasvacuolesbyaquaticbacteriaprovidesamechanismforadjustingthebuoyancyofthecell.Manyaquaticcyanobacteriausetheirgasvacuolestomoveupanddowninthewatercolumn.第五十九页,共九十二页。2.6ComponentsexternaltothecellwallFlagellaFimbriaeandpiliCapsulesandslimelayers第六十页,共九十二页。Motilityallowsthecelltoreachdifferentregionsofitsenvironment.Inthestruggleforsurvival,movementtoanewlocationmaymeanthedifferencebetweensurvivalanddeathofthecell.But,asinanyphysicalprocess,cellmovementiscloselytiedtoanenergyexpenditure,andthemovementofflagellaisnoexception.
Manyprokaryotesaremotile,andthisabilitytomoveindependentlyisusually
duetoaspecialstructure,
the
flagellum
(plural,flagella).
第六十一页,共九十二页。第六十二页,共九十二页。Fourbasictypesofflagellararrangementsa.monotrichousb.amphitrichousc.
lophotrichousd.peritrichous第六十三页,共九十二页。Flagellaarearrangeddifferentlyondifferentbacteria.Inpolarflagellationtheflagellaareattachedatoneorbothendsofthecell.Occasionallyatuft(group)offlagellamayariseatoneendofthecell,anarrangementcalledlophotrichous.Inperitrichousflagellationtheflagellaareinsertedatmanyplacesaroundthecellsurface(perimeans"around").Thetypeofflagellation,polarorperitrichous,isoftenusedasacharacteristicintheclassificationofbacteria.第六十四页,共九十二页。
TheflagellumofaGram-negativebacterium第六十五页,共九十二页。Thefilamentofbacterialflagellaiscomposedofsubunitsofaproteincalled
flagellin.
Thebaseoftheflagellumisdifferentinstructurefromthatofthefilament.Thereisawiderregionatthebaseoftheflagellumcalledthe
hook.Thehookconsistsofasingletypeofproteinandfunctionstoconnectthefilamenttothemotorportionoftheflagellum.
The
basalbody
isanchoredinthecytoplasmicmembraneandcellwall.Thebasalbodyconsistsofasmallcentralrodthatpassesthroughasystemofrings.第六十六页,共九十二页。Ingram-positiveBacteria,whichlacktheouterlipopolysaccharidelayer,onlytheinnerpairofringsispresent.SurroundingtheinnerringandanchoredinthecytoplasmicmembraneareapairofproteinscalledMot.Theseproteinsactuallydrivetheflagellarmotorcausingatorquethatrotatesthefilament.Afinalsetofproteins,calledtheFliproteinsfunctionasthemotorswitch,reversingrotationoftheflagellainresponsetointracellularsignals.Ingram-negativeBacteria,anouterringisanchoredinthelipopolysaccharidelayerandanotherinthepeptidoglycanlayerofthecellwall,andaninnerringislocatedwithinthecytoplasmicmembrane.第六十七页,共九十二页。Themovementofaprocaryoticflagellumresultsfromrotationofitsbasalbodyandissimilartothemovementoftheshaftofanelectricmotor.Bacterialcellscanalterthespeedanddirectionofrotationofflagellaandthusarecapableofvariouspatternsofmotility.第六十八页,共九十二页。Fimbriae
canbeevenlydistributedovertheentiresurfaceofthecell.Theycannumberanywherefromafewtoseveralhundredpercell.Fimbriaeallowacelltoadheretosurfacesincludingthesurfacesofothercells.
Piliareusuallylongerthanfimbriaeandnumberonlyoneortwopercell.PilifunctiontojoinbacterialcellspriortothetransferofDNAfromonecelltoanother.Fpilus第六十九页,共九十二页。Manyprokaryotescontainacellsurfacelayercomposedofatwo-dimensionalarrayofprotein.TheselayersarecalledS-layers.S-layershavebeendetectedinrepresentativesofvirtuallyeveryphylogeneticgroupingofBacteriaandarenearlyuniversalamongArchaea.InsomespeciesofArchaeatheS-layerisalsothecellwall.ParacrystallineSurfaceLayers(S-Layers)第七十页,共九十二页。ThemajorfunctionofS-layersisunknown.Astheinterfacebetweenthecellanditsenvironment,itislikelythatincellsthatproducethemtheS-layeratleastfunctionsasanexternalpermeabilitybarrier,allowingthepassageoflow-molecular-weightsubstanceswhileexcludinglargemolecules.第七十一页,共九十二页。2.7BacterialEndosporesCertainspeciesofbacteriaproducespecialstructurecalledendospores.Theyareveryresistanttoheatandcannotbedestroyedeasily,evenbyharshchemicals.Endosporesarealsoresistanttootherharmfulagentssuchasdrying,UVradiation,acidsandchemicaldisinfectants.第七十二页,共九十二页。SporulatingcellCentralcoreCortexSporecoat/membraneexosporiumMicrographofaendosporeVegetativecell第七十三页,共九十二页。第七十四页,共九十二页。Manyprokaryoticorganismssecreteontheirsurfacesslimyorgummymaterials.Avarietyofthesestructuresconsistofpolysaccharide,andafewconsistofprotein.Thetermscapsuleandslimelayerarefrequentlyusedtodescribepolysaccharidelayers.Demonstrationofthepresenceofacapsuleisusuallybymeansofnegativestaining
CapsulesandSlimeLayers第七十五页,共九十二页。LowpHvalue,andcontainssmallacid-solublesporeproteins(SASPs)StructureofDPA
PropertiesofEndosporeandItsResistanceContainingabundantDPA(dipicolinicacid)whichiscombinedwithcalciumions.Lowerwatercontent–only10-30%ofthewatercontentofthevegetativecell.第七十六页,共九十二页。Sporulationinvolvesaverycomplexseriesofeventsincellulardifferentiation.Bacterialsporulationdoesnotoccurwhencellsaredividingexponentiallybutonlywhengrowthceasesowingtotheexhaustionofanessentialnutrient.Thus,cellsofBacillusceasevegetativegrowthandbeginsporulationwhenakeynutrientsuchasthecarbonornitrogensourcebecomeslimiting.EndosporeFormatiom第七十七页,共九十二页。
Stagesinendosporeformation1.Axialfilamentformation2.Septumformatiom3.Engulfmentofforespore4.Cortexformation5.Coatsynthesis6.Completionofcoatsynthesis,Increaseinrefractilityandheatresistance7.Lysisofsporangium,sporeliberation第七十八页,共九十二页。第七十九页,共九十二页。1.Activation–
Usuallyresultsfromtreatmentslikeheating.2.Germination–
Breaksspore’sdormantstate.Thisprocessischaracterizedbysporeswelling,lossofresistancetoheatandotherstresses,lossofrefractilityandincreaseinmetabolicactivity.3.Outgrowth–
Thesporeprotoplastmakesnewcomponentsanddevelopsoncemoreintoanactivebacterium.SporeGermination第八十页,共九十二页。SeveralBacillusspecies,mostnotablyB.popilliaeandB.thuringiensis,produceintracellularcrystalsoftoxicglycoproteinswhentheysporulate.ToxiccrystalsParasporalCrystal(Spore-companionedcrystal)第八十一页,共九十二页。2.8ComparisonofTheProkaryoticandEucaryoticCellPropertiesProkaryoteEukaryotePhylogeneticgroupsBacteria,ArchaeaEukarya:Algae,fungi,protozoa,plants,animalsSizeGenerallysmall,usually<2µmindiameterUsuallylarge,2to100µmindiameterNuclearmembranesAbsentPresentNucleolusAbsentPresentDNANotmitosisMitosis;mitoticapparatuswithmicrotubularspindle第八十二页,共九十二页。PropertiesProkaryoteEukaryoteCytoplasmicmembraneUsuallylackssterols;hopanoidsmaybepresentSterolsusuallypresent;hopanoidsabsentRibosomes70sinsize80s,exceptforribosomesofmitochondriaandchloroplastswhichare70sMembranousorganellesAbsentSeveralpresentRespirationsystemPartofcytoplasmicmembrane;mitochondriaabsentInmitochondria;hydrogenosomesincertainanaerobicspeciesCellwallsPresent(inmost),composedofpeptidoglycan(bacteria),protein,otherpolysaccharides,glycoprotein(Archaea)Presentinplants,algae,fungi,usuallypolysaccharide;absentinanimals,mostprotozoa第八十三页,共九十二页。TheirDNAisassociatedwithhistoneproteins.membrane-boundedorganelles9+2typeflagellaEucaryoticCell第八十四页,共九十二页。Thechiefdistinguishingcharacteristicsofprocaryoticcellsare:1.Theirgeneticmaterial(DNA)isnotenclosedwithinamembrane.2.Theylackothermembrane-boundedorganelles.3.TheirDNAisnotassociatedwithhistoneproteins.4.Theircellwallsalmostalwayscontainthecomplexpolysaccharidepeptidoglycan.5. Theyusuallydividebybinaryfission.Duringthisprocess,theDNAiscopiedandthecellsplitsintotwocells.
第八十五页,共九十二页。Summary1.Prokaryoticgeneticmaterialislocatedinanareacalledthenucleoidandisnotenclosedbyamembrane.2.Mostbacteriahaveacellwalloutsidetheplasmamembranetogivethemshapeandprotectthemfromosmoticlysis.Bacterialwallsarechemicallycomplexandusuallycontain
peptidoglycanormurein.第八十六页,共九十二页。3.Bacteriaoftenareclassfiedaseithergrampositiveorgramnegativebasedondifferencesincellwallstructureandtheirresponseto
Gramstain
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