植物生理与分子生物学：叶绿体发育

叶绿体发育叶绿体发育质体的种类与功能质体的起源质体的分裂叶绿体的发育质体的种类与功能硅藻、眼虫纤毛虫三重内共生二重内共生一些甲藻质体的起源—内共生KonstantinMereschkowsky于1905年最先提出叶绿体是由原先的内共生体形成的这一想法，随后IvanWallin于1920年提出了对线粒体的相同想法。硅藻、眼虫纤毛虫三重内共生二重内共生一些甲藻含有类似于细菌的DNA与遗传密码。被两层或更多的膜所包被。细胞器的大小与细菌相当，通过二分分裂方式繁殖。内部结构和生物化学特征，如70S（55S）核糖体和类囊体。古虫界变形虫界后鞭毛生物有孔虫界囊泡藻界泛植物界光合生物的多样性与光合机构的统一性高等植物中有4300个蛋白质是来自蓝细菌。通过预测大约有3000个核基因产物定位于叶绿体中。蛋白质组学目前只能检测到1500个左右。植物叶绿体的分裂AlternationinnumberandmorphologyofchloroplastsinarcmutantsWTarc6arc1arc11Arc3arc5arc11arc6Marrisonetal.,1999PlantJ.AccumulationandReplicationofChloroplasts叶绿体随细胞生长而分裂Marrisonetal.,1999PlantJ.核基因表达及其产物转运叶绿体基因表达核与叶绿体相互作用叶绿体发育核基因编码的叶绿体蛋白主要通过TOC-TIC途径进入叶绿体，少量的蛋白通过分泌途径。TOC-TIC转运机构toc33蛋白转运受到多层次调控TransloconabundanceTransloconisoformsRedoxconditionsandCa2+

蛋白质转运调控Lingetal.,2012Science叶绿体基因表达Arabidopsis154,478bp4ribosomalRNAgenes37tRNAgenes87proteincodinggenesNoordallyetal.,2013ScienceWoodsonetal.,2013PlandJ.叶绿体基因特性NEP与PEP的相互调控根据表达特性叶绿体基因分为三类•TranscribedsolelybyPEP(classI)•TranscribedbyPEPandNEP(classII)•TranscribedsolelybyNEP(classIII)ClassIClassIIClassIIIpsaAatpBaccDpsbBclpPrpl33/rps18psbEndhBycf2petBndhFrpoBndhArps16rps14rrnrbcLatpIpsbApsbD叶绿体rrn操纵子的转录与成熟17Nucleasesinplastids:Endonucleases,ExonucleasesRNEandRHON在rRNA成熟中的作用Stoppeletal.,2012Nuc.AcidRes复杂的转录后调控ChloroplastRNARNAsplicingRNAeditingNucleusPPR70SRNAtrimmingPentatricopeptiderepeatproteinPPRproteinsPPRproteinactionRNAremodelingactivityofagenericPPR

.Proteincouldexposeormaskcis-elementsfortranslation,RNAsplicing,RNAcleavage,orRNAediting.MechanismoftranslationalactivationbyPPR10.PPRproteinscanstabilizeRNAsanddefinethepositionsofprocessedRNAterminibyblockingexoribonucleases.复杂的转录后调控叶绿体蛋白质合成叶绿体的70S核糖体由50S的大亚基和30S小亚基组成，30S小亚基由16SrRNA和25个RPS蛋白组成，50S的大亚基由23S，5S，4.5SrRNA和33个RPL蛋白组成。植物中有6个特有的蛋白被称之为PSRP蛋白，其中4个在30S，2个在50S。30Sand50SRibosomeassemblyinBacteriaTilleretal.,2012,PlantJ.敲除PSRP2与PSRP6不影响植物生长发育、光合作用。SeedlingLethalityoftheΔrps15/Δrpl33MutantsisRescuedbyElevatedGrowthTemperatures叶绿素合成EspinedaCEetal.PNAS1999;96:10507-10511Chlorophyllaoxygenase(CAO)mutants类胡萝卜素膜脂合成质体与核的信号交流AnterogradeSignalingWoodsonandChory,2008NatureReviewGeneticsRetrogradeSignalingcch:Conditionedchlorina;cs

chlIcch:Conditionedchlorina;cs

chlI叶绿体发育的研究叶绿体发育—白花苗、渐渐转绿、斑叶、条斑等Sakamoto,W.,(2003).GenesGenet.syst78:1-9.Wang,Q.etal.(2004).PlantPhysiol.136:3594-3604.thf1VAR1(FtsH5),VAR2(FtsH2):metalloprotease,ATPaseassociatedwithvariouscellularactivities.IM(Immutans),plastidterminaloxidase(PTOX).THF1(thylakoidformation1/psb29,unknownprotein).LeafvariegationmutantsinArabidopsisLeafvariegationmutantsWhyischloroplastdevelopmentarrestedinsomesectorsbutnotinothersinthesamegeneticbackground?Chloroplastdevelopmentinthemutantsissensitivetodevelopmentalandenvironmentalcues.ChlorophyllautofluorescenceHypothesesforleafvariegationPhotosyntheticredoximbalanceFtsHthresholdmodelImbalancebetweenproteinbiosynthesisanddegradationinplastidsPhotosyntheticredoximbalanceRossoetal.,Plantcell,2009Ultrastructureofetioplastsinthf1andvar2WTthf1

thf1thf1

var2var2Wuetal.,(2011)JIPBPLBPhotosyntheticredoximbalanceFoudreeetal.,(2012)FrontiersinPlantSci.

Im(PTOX)functionsasasafetyvalvetopreventover-oxidationofthePQpoolinhighlightstress.immutantsaccumulatePhytoenebutnotcarotenoids.AthresholdmodelSubunits:TypeA(FtsH5/VAR1,FtsH1),TypeB(FtsH2/VAR2,FtsH8).TorepairD1proteinatthePSIIreactioncenter.AthresholdofFtsHactivityisrequiredforchloroplastdevelopment.Thethresholdisdynamicandmayreflectthephysiologicalconditionsofplantcells.LeafvariegationisrescuedbyanincreaseinFtsHlevels.Yuetal.,(2004)PlantJ.Yuetal.,

(2005)PlantPhysiol.

Zaltsmanetal.,(2005)PlantCell.Over-expressingFtsH8rescuesvar2WTvar2

FtsH8/var2Yuetal.,(2004)PlantJ.ImbalancebetweenproteinsynthesisanddegradationinplastidsMiuraetal.,(2007)PlantCell19:1313-1328.Liuetal.,(2010).BMCPlantBiol10:287.Proteintranslation:Fug1(cpIF2),Sco1(cpEF-G)rRNAprocessing:SVR1(pseudouridinesynthase)chloramphenicolspectinomycinIntegratedhypothesisDefectsinchloroplasttranslationprolongstheprocessofchloroplastbiogenesis,whichmayreduceathresholdofFtsHactivity.Reductionsinchloroplasttranslationturnonretrogradesignalingfromplastidstothenucleus,resultinginalteredexpressionofnucleargenesandalsoalteredchloroplastproteomethatallowsdevelopingplastidstocompensateforalackofVAR2.Wuetal.,unpublishedIdentificationofsuppressor7ofthf1(sot7)(47-2)SOT7encodestheR4subunitofClpproteaseWuetal.,unpublishedClpproteaseand26SproteosomesystemsChloroplastClpPRT1T2Mogketal.,(2007)TrendsinCellBiol.clpmutantsareseedling/embryolethalorvirescentKim,J.,etal.,(2009).PlantCell21:1669-1692.Olinares,P.D.,etal.,(2010).BBAMutationsinothersubunitssuppressthf1variegation,andclpr4suppressesvar2variegationWuetal.,unpublishedProteomicsapproachProteinsiTRAQlabelingMIXiTRAQlabeledpeptidesTrypticPeptidesLC-MS/MSandquantificationofproteins

1598P1545G

clpR4-3thf1proteomeiscorrelatedwiththatofclpR4Spearman:0.8180.5390.4811629P1598P31P105P52G

1545G1313C(85%)Wuetal.,unpublishedclpR4thf1chloroplastproteomeisdominatedbyclpR4thf1clpR422thf19clpR42362261575518Mis-regulatedproteinsp≤0.05,fold-change≥1.5Wuetal.,unpublishedSubunitsofClpRringaccumulatelessinclpr4clpR4-3/WTthf1/WTclpR4thf1/WTWuetal.,unpublishedManychaproneproteinsincreaseinclpr4Wuetal.,unpublishedManyribosomalproteinsdown-regulatedinclpR4

clpR4-3/WTthf1/WTclpR4thf1/WTWuetal.,unpublishedKnockingoutPRPL11suppressesleafvariegation(thf1prpl11)Wuetal.,unpublishedprpl11rescuesvar2leafvariegationWuetal.,unpublishedChloroplastrRNAlevelscoordinatelydecreaseWuetal.,unpublishedEffectofClpR4mutationonbiologicalprocessesClpR4mutationFoldingstressAntioxidativedefenseCalvin-BensoncycleChaperonePSIIbiogenesisRibosomalproteinsrRNAsPlastidtranscriptionABA,JA,starchChloroplastdevelopmentslowsdownthf1thf1Namethf1/WTclpR4-3/WT47-2/WTB1B2B1B2B1B2THF1-45.78-50.35-1.73-1.70-57.81-58.99FTSH2-36.33-49.11-1.94-2.94-21.63-50.38FTSH8-20.87-37.74-1.98-2.73-12.91-36.33FTSH5-8.13-8.33-13.94-22.23-11.62-12.16PTAC16-8.78-2.501.568.831.4311.26FKBP13-2.19-6.923.1615.653.4414.45PRXQ-2.47-2.41-2.35-2.09NA-2.84DHAR3-1.85-4.11-2.66-2.39NA-2.82DUF1350-2.49-2.45NA-7.22-3.49-6.67DUF1350-2.56-2.64NANA-5.26NACCD4-4.10-2.98NANANANAASP5-2.65-5.96NANANANAPsaC4.471.55NANANANAPSRP21.502.05NANA-1.85NAPRXIIE2.013.18NANANANAENH12.282.03NANANA-2.60CPN21/GroES1.5111.50NANANANACP12-12.5111.03NANANANAWuetal.,unpublishedFtsHaccumulationinclpR4thf1ishigherthaninthf1ItmaynotbetruethatleafvariegationisrescuedbyareducedthresholdofFtsHactivity.Wuetal.,unpublishedLeafvariegationZhangetal.(2009)PlantJ.Wuetal.(2013)Plan