全 文 ：第 25卷 第 3期 植 物 研 究 2005年 7月Vol.25 No.3 BULLETINOFBOTANICALRESEARCH July, 2005
Foundationitem:SupportedbytheStateKeyBasicResearchandDevelopmentPlanofChina(G19990160)
Authorintroduction:ZuYuangang(1954—), Ph.D., Professor, majorinplantscience.
Receiveddate:2005-01-07
植物叶片原生质体分离的可能机制
祖元刚 于景华 唐中华 郭晓瑞 张宇亮 孟庆焕
(东北林业大学森林植物生态学教育部重点实验室 , 哈尔滨 150040)
摘 要 分析了植物叶片在分离液环境中形成原生质体的过程 ,文中提出 ,分离液配方中的酸性
物质使植物叶片处于酸性环境中并导致植物正常细胞首先发生细胞壁酸性降解 ,随后出现原生质
体脱离细胞壁进入分离液 ,继而又进一步发生质膜的酸性降解 ,使细胞核和细胞器进入分离液中 ,
最终分离液中的细胞器以细胞核为中心进行细胞器重组 ,最后产生外貌形态一致的新的原生质
体 。植物细胞壁和质膜是植物细胞的包被系统。植物细胞包被系统的酸性降解使植物细胞器重
组并产生新的原生质体成为可能。
关键词 植物叶片;细胞壁酸性降解;原生质体
Possiblemechanisminvolvedwithisolationprotoplastofleaf
ZUYuan-Gang YUJing-Hua TANGZhong-Hua GUOXiao-Rui ZHANGYu-Liang MENGQing-Huan
(KeyLaboratoryofForestryPlantEcology, MinistryofEducation, NortheastForestryUniversity, Harbin 150040)
Abstract Theprocessandmechanismofleafprotoplastformationafterincubationtocelwaldigesting
solutionwasinvestigated.Theacidmaterialinthedigestingsolutionprovidedtheacidenvironmentfor
leavesandledtotheoccurrenceofaciddegradationofcelwal, theseparationbetweencelwalwith
protoplastandtheescapeofprotoplastfromcelwalintosolution.Thecytoplasmmembraneinsidethe
digestingsolutionwasdegradedbytheacidenvironmentandthisaccountedfortheescapeofcelnucleus
andorganelesfromprotoplast.Indigestingsolution, therecombinationoforganelessuroundingnucle-
arandsubsequentformationofnewprotoplastwithunanimousshapetookplace.Theseresultsshowed
thatplantcelwalandcytoplasmmembraneisthecoatsystemofplantcel.Theaciddegradationof
plantcelcoatsystemmakesrecombinationofcelorganandproductionofnewprotoplastpossible.
Keywords leaf;aciddegradationofcelwal;protoplast
Celsofprimaryplanttisuespossesscelulosic
walswithapectin-richmatrix, themiddlelamela,
joiningadjacentcels.Thelivingcytoplasmofeach
cel, boundedbytheplasmamembrane, constitutes
theprotoplast[ 1] .Plantcelwalandprotoplastare
thecelcoatsystemanditmakescrucialrolesduring
celcycle.Ifalargenumberofprotoplastsarere-
quired, whichisthenorm, enzymaticdigestionof
sourcetissuesisessential[ 1 ～ 5] .Thewalofplantleaf
celincubatedtothedigestingsolutionfirstlywasde-
gradedduetotheenvironmentandsubsequentescape
ofplantprotoplast("naked"cel)fromcelwalwil
occur.Theseprocessesaccountforthedisorganization
ofcels.Andthefurtherinteractionofacidmaterial
inthedigestingsolutionwiththeescapedprotoplast
wildissectthecytoplasmmembraneofprotoplast, re-
sultingintheorganelesuchasnucleusandchloro-
plast, etc.tobereleasedfromprotoplast.Chloro-
plast, mitochondrialandotherorganelesdistributed
insolutionwilaggregategradualyandrecombine
newprotoplastwithunanimousappearance.
Duringthecourseofceldegradationandpro-
ductionofprotoplastescapedfromcelwal, theacid
environmentandvariouskindsofdegradingenzymes
servedthemostimportantfunctions.Someenzymes
areimplicatedtomaintenanceofosmoticpotentialin-
sideandoutsidetheprotoplastwiththesolublesugar,
suchascelulaseandpectinaseservingtodegradethe
celwal.Inthedigestingsolution, thepHvaluewas
5.8andinsuchacidenvironment, thepectinasewil
catalyzepectintodecomposeandthecelulasehydro-
lyzedthecelulosemoleculeinthecelwalintothe
glucose[ 6 ～ 10] .Thestructureofcelwalwilbede-
stroyedbythewayofenzymaticlydegradation.Some
celwalstotalydisappearfinaly, withprotoplast
separatedfromcelwalforrecombinationofentire
celincludingcelwalandprotoplast.Thiscourse
wascaledinthisresearchaciddegradationofcel-
coat-system.
ItisobviousthatpHvalueofmicroenvironment
insidethecelwalishigherthanithatindigestingso-
lutioninthenormallifestate.Thereislitlepossibili-
tyoftheaciddegradationofplantcelinnormalin-
stanceandthiswilkeepthestabilityandintegrityof
cel, consequentlythenormaltransportationofexter-
nalsignaltoinsidenuclearthroughcelwalandcyto-
plasmmembrane[ 11, 12] andcelcycle.However, after
acidenvironmentandalkindsofenzymesindiges-
tingsolutionmakethecelwaldegrade, andplant
protoplastreleasedfromcelwalcoatdirectlyliein
acidenvironment.Itmayimpelprotoplasttopresent
furtheraciddegradation, andformthenewprotoplast
withunanimousshapefinaly.
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2733期 祖元刚等:植物叶片原生质体分离的可能机制