dominant是什么意思inant在线翻译读音例

JournalofIntegrativePlantBiology2006,48(4):447−452

Received21Mar.2005Accepted15Aug.2005

SupportedbytheNationalNaturalScienceFoundationofChina(90208001)

andtheKnowledgeInnovationProgramoftheChineseAcademyofSci-

ences(KSCX2-SW-306).

*Authorsforcorrespondence.E-mails:and

.

;

GeneticAnalysisandMappingoftheDominant

DwarfingGeneD-53inRice

Li-RongWei1,Ji-ChenXu1,Xiao-BoLi1,QianQian2*andLi-HuangZhu1*

(uteofGeneticsandDevelopmentalBiology,theChineseAcademyofSciences,Beijing100101,China;

eyLaboratoryofRiceBiology,ChinaNationalRiceResearchInstitute,Hangzhou310006,China)

Abstract

ThedwarfinggeneD-53isoneofafewdominantgenesfordwarfinginrice(OryzasativaL.).Inthepresent

study,ourgeneticanalysisconfirmedthatmutantcharacteristicsincludingdwarfing,profusetillering,thin

uredthelengthofeach

internodeofKL908,aD-53-carryingline,tion,we

measuredelongationofthesecondsheathand

-amylaseactivityintheendosperm,andwecharacterized

KL908asadwarfmutantthatwalarge

F

2

populationobtainedbycrossingKL908withawild-typevariety,NJ6,theD-53genewasmappedtotheterminal

regionoftheshortarmofchromosome11,withonesimplesequencerepeatmarker,Ds3,co-segregating,and

theother,K81114,located0.6cMaway.

Keywords:dominantdwarfgene;geneticanalysis;geneticmapping;rice.

WeiLR,XuJC,LiXB,QianQ,ZhuLH(2006).rat

PlantBiol48(4),447−452.

Dwarfvarietiesofcropscannotonlyincreaseresistanceto

lodgingonaccountoftheirshortstature,butalsoproducehigh

yields,withincreasesingrainweight(Borlaug1983;Galeand

Youssefian1985;Evans1993).Rice,agloballyimportantcrop,

hasalargenumberofdwarfmutants(Kinoshita1995).However,

-irradiatingrice

varietyNL8,Iwataandhiscolleaguesobtainedadominantdwarf

mutantwithprofusetillers,andmappeditsdominantgene,D-K-3,

tothe8thlinkagegroupofrice(Iwataetal.1977,1978).Atthat

time,the8thlinkagegroupwasthoughttobelongtothe9thchro-

mosome(IwataandOmura1984),butKinoshita(1984),whofor-

mallyrenamedthegeneD-53,laterremappedD-53tochromo-

some11(Kinoshita1991).Sincethenonlyafewstudiesofthis

l.(1988)and

Zhuetal.(1995)carriedoutsomeinvestigationsoftheshortculm

r,nostudyhasbeen

carriedouttoconfirmthelocusofD-53byusingmolecularmarkers.

Studiesofricedwarfcharacteristicshaveinvolved

morphological,cytologicalandphysiologicalanalysisofsome

(1977)categorizedricedwarfmutants

intosixgroupsonthebasisoftheelongationpatternofintern-

egroups,dn-typemutantshaveeachinter-

nodeuniformlyshortened,whereasthedm-typeandthesh-type

mutantshavespecificallyreducedlengthofthesecondinternode

andthefirstuppermostinternode,-typemutants,

thefourthinternodeisrelativelylonger,whereasthefirstintern-

logically,dwarfricephenotypesmay

resultfromchangesinvariousmechanisms(Reiteretal.1993;

Takahashietal.1995;LiandChory1997;Azpirozetal.1998;

Multanietal.2003).Gibberellicacid(GA),oneofthemostimpor-

tantplantendogenoushormones,山围故国周遭在 isindispensableforthegrowth

ofplants(Hooley1994;Rossetal.1997).Eitherabnormalsynthe-

sisofGAordisruptionoftheGAsignaltransductionpathwaycan

leadtointerruptionofelongationoftheinternodes,ultimatelyre-

sultingindwarfism(Ikedaetal.2001;Itohetal.2001).Onthe

basisoftwoGA-mediatedprocesses,namelyelongationofshoots

andproductionof-amylaseactivityintheendosperm,Mitsunaga

etal.(1994)classifiedricedwarfmutantsintothreegroups:T,D,

e,theTgroupcomprisesGA-deficientmutants,

andtheDgroupGA-insensitivemutants,whereastheEgroup

448JournalofIntegrativePlantBiologyVol.48No.42006

comprisesmutantsthatareneitherGA-deficientnorGA-

insensitive.

Anumberofdwarfinggenesinricehavebeencharacterized

bymap-basedcloning(Ashikarietal.1999;Morietal.2002;Sasaki

etal.2002;Hongetal.2003),butthesecloneddwarfgenesare

ore,itisimportanttodeterminetheparticular

resent

study,weinvestigatedthemorphologicalandphysiologicalchar-

acteristicsofaD-53carryingline,KL908,andcarriedoutgenetic

analysisthroughacrossbetweenKL908andNJ6(anormalin-

dicavariety).虚怀若谷 安之若素 Furthermore,wepreciselymappedthegenetothe

terminalregionoftheshortarmofricechromosome11byusing

molecularmarkers.

Results

PhenotypicandgeneticanalysisofthemutantKL908

ThedwarfmutantKL908hasshortstems(Figure1A),whichare

onaverage70cminheight,muchshorterthanthoseofthevariety

NJ6,1andFigure1Blistthe

lengthofeachinternode出尔反尔 ofKL908incomparisonwiththatofNJ6.

TherelativelengthsofeachinternodetotheculminKL908and

NJ6areshowninFigure2,demonstratingthateachinternodeis

evenlyshortenedinKL908,ore,thedwarf

mutantKL908isdn-typeaccordingtothecategoriesofTakeda

(1977).

Inadditiontoitsdwarfcharacteristics,weexaminedotherphe-

notypiccharacteristicsofKL908,suchasitsprofusetillers,small

lernumbersofindividualplantsof

KL908andNJ6duringtheheadingtimearepresentedinTable2.

AsshowninFigure1B,thestemsofKL908areobviouslythinner

thanthoseofNJ6,andthepaniclesofKL908aresmallerthan

theF

2

populationfromthecrossbetween

KL908andNJ6tocarryoutco-segregationanalysisofthese

heindi-

vidualsoftheF

2

population,thenumberofmutant-typeindividuals

withdwarfism,profusetillers,smallpaniclesandthinstemswas

323,andthecorrespondingnumberforthewild-typeindividuals

ioofmutantphenotype:wildtypewasapproxi-

mately3

:

1(2=0.7264),confirmingthatD-53isapleiotropic

dominantgene.

ypeofthericemutantKL908.

(A)OverallrepresentationofthephenotypesofthevarietyNJ6(a)

andthemutantlineKL908(b).

(B)Comparisonofthelengthsandthethickness昆的拼音 ofallinternodesof

NJ6(left)andKL908(right).

P,panicle;I–V,therespectiveinternodesfromtoptobottom.

elengthofeachinternodeoftheparentsafterhead-

ing

LengthKL908(cm)NJ6(cm)

The1stinternode24.8(1.39)42.2(2.46)

The2ndinternode11.6(1.34)29.6(1.18)

The3rdinternode7.0(0.92)22.4(1.69)

The4thinternode5.4(0.66)18.1(0.48)

The5thinternode2.6(0.54)6.4(1.36)

Total51.3(1.64)118.6(4.69)

GeneticMappingofRiceDwarfGeneD-53449

RelationshipbetweendwarfingandGAinKL908

Toinvestigatethepossibleconnectionbetweenthedwarfing

mechanismandGAinKL908,weexaminedtheeffectsofshoot

elongationandendosperm-amylaseactivityinduction(bothof

whichareGA-promotedphysiologicalprocesses)byGAapplica-

tiontotheKL908mutant(Mitsunaga1994).AsshowninTable3,

theratio(1.53)ofthelengthsofthesecond-leafsheathwithand

withoutpriorapplicationofGAinKL908wasnotsignificantly

differentfromthatinNJ6(1.56),suggestingthatdwarfisminthe

ine

othereffectsofGAonD-53plants,agarplateassaysforamy-

samepatternsof-amylaseactivityinductionbyGAwereob-

servedinbothNJ6andKL908half-seeds(Figure3),whichdem-

onstratesthattheD-53mutantisnotaGA-insensitivedwarf.

Therefore,wepresumedthatthedwarfisminKL908isnotasso-

ingtothesystemofMitsunaga

etal.(1994),thedwarfmutantKL908belongstogroupE,those

thatareneitherdeficientnorinsensitivetoGA.

MappingoftheD-53geneusingmolecularmarkers

Knownsimplesequencerepeat(SSR)markersonthericechro-

mosome(McCouchetal.2002)wererandomlyselectedforpoly-

morphismsurveysbetweenKL908andNJ6,resultingin46posi-

tivemarkersthatwerethenappliedto32randomlyselectedF

2

individualswiththewild-typephenotypeforsegregationanalysis.

Ofthesemarkers,RM167,RM3668andRM20,allofwhichare

locatedontheshortarmofchromosome11,showedcloseasso-

ranalysisusing150wild-type

individualsshowedthatthegeneticrecombinationfrequencies

betweentheD-53geneandthemarkersRM167,RM3668and

RM20were27.0%,7.6%and5.6%,ore,the

targetgenewaslocatedontheshortarmofthechromosome11.

SincetherecombinantsforRM20wereincludedinthosefor

RM3668andtherecombinantsforRM3668wereincludedinthose

forRM167,wemappedtheD53genetotheregionfromtheRM20

markertotheterminalregionoftheshortarmofthe11thchromo-

some(Figure4).

FinemappingofthedominantdwarfgeneD-53

Intheregionbetweenthedistalendoftheshortarmofchromo-

some11andSSRmarkerRM20,wedetected14newSSRse-

quencesbasedonpublicDNAsequences(.

).Ofthesemarkers,K81114andDs3werepolymorphicbe-

rimersequencesareshown

lernumbersofNJ6andKL908duringheadingtime

LineAverage

NJ6110810.4

KL9554847504249.8

uctionof-amylaseactivity.

+GA

3

,appliedGA

3

intheplate;−GA

3

,notappliedGA

3

intheplate.

onoftheD-53geneintheshortarmofchromosome

11.

450JournalofIntegrativePlantBiologyVol.48No.42006

l,552individualswiththerecessivephenotypein

theF

2

populationweresurveyedbyusingthenewSSRmarkers.

ForthemarkerK81114,sevenrecombinantswerefound,and

tion,themarker

eanalysiswith

Mapmaker/Exp3.0softwareshowedthatthemapdistancebe-

tweenthetargetgeneD53andthemarkerK81114was0.6cM

(Figure4).

Discussion

Sincetheso-called“greenrevolution”,manydwarfricemutants

havebeenfound(Kinoshita1995).Butonlyafewstudiesregard-

ghZhuet

al.(1995)geneticallyanalyzedthedwarfingtraitofKL908,they

didnotdiscusstherelationbetweendwarfismandothermutant

gco-segregationanalysis,wecametotheconclu-

sionthatallthemutantcharacteristicsofKL908,includingdwarfism,

profusetillering,smallpaniclesandslenderstalks,areallcon-

trolledbythedominantgene,D-53.

AccordingtoTakeda’ssystemofcategorizingofdwarfrice

mutants,rfisminKL908

ly,the

dwarfingmechanismsofanumberofricedwarfmutantshave

tud-

ieshavefoundthatthedwarfingmechanismsindifferentdwarf

mutantsarevarious(Ashikarietal.1999;Morietal.2002;Sasaki

etal.2002;Hongetal.2003).First,thed1genewasfoundto

encodeafunctionalGTP-bindingproteinthatisinvolvedinGA

signaltransduction(Ashikarietal.1999).Thereafter,asemi-dwarf

gene,sd1,theso-called“greenrevolution”geneinrice,was

foundtobedefectiveinencodinganactiveGAsynthesisenzyme

(Sasakietal.2002;Spielmeyeretal.2002).Furthermore,two

malformeddwarfmutantricegenes,d61andd2,werecharacter-

izedascausingfailureintheperceptionandsynthesis,

respectively,ofbrassinolide(BR)(Yamamuroetal.2000;Honget

al.2003).TheD-53geneisoneofonlyafewdominantdwarf

genesinrice(汪曾祺散文摘抄 KohandHeu1993).Asmentionedearlier,thechar-

acteristicsconferredbytheD-53genearenotonlydwarfism,but

alsoprofusetillers,siologi-

calstudyexcludedonlythepossibilityoftheinvolvementofGAin

thedwarfingmechanismofKL908,butitispossiblethatanother

gand

identificationoftheD-53geneisneededtoelucidatethispoint,

andtodeterminewhatdefectcausesthedeformeddwarfpheno-

typeinthemutantplants.

Inthepresentstudy,wild-typeindividualsinalargeF

2

popula-

tionwereusedforsegregationanalysisoftheD-53geneandthe

polymorphicmarkers,becausetherecessivewild-typ相看白刃血纷纷 epheno-

tenthegapbetweenthetar-

getedgeneanditsflankingmarkers,moreSSRmarkerswere

exploredanddesignedbyreferringtoricegenomesequencesin

thatregion().Sowecouldfinallyidentify

newSSRmarkersthatweremorecloselylinkedtotheD-53gene,

resultingintheidentificationofonemarker,Ds3,whichco-segre-

gatedwiththegeneandtheother,K8111,beingonly0.6cM

unately,wefailedtoidentifyanothermarkeronthe

othersideofD-53inordertomorepreciselydefinetheD-53-

ownthatthesequenceoftheD-53-

containingregiononchromosome11isaduplicationofthese-

quenceonchromosome12(Wuetal.1998).Thismaybeareason

whywewereunabletofindmoremarkerstomapthetargetgene

betweentwocloselylinkedmarkersinthisterminalregionofthe

sly,tofinallyclonetheD-53

gene,itisnecessarytoconstructalargersegregationpopulation

andtoexploremoremolecularmarkers.

MaterialsandMethods

Plantmaterials

KL908,aD-53carryingline,derivedfromtheoriginalmutantob-

tainedby-irradiatingthejaponicavarietyNL8(Iwataetal.1977),

6isatallindicavariety

2

mappingpopulation,

consistingof2215individuals,wasderivedfromacrossbe-

rice(OryzasativaL.)plantswere

ading,whenalltheindividuals

andtheparentalplantsweretasseled,weinvestigatedthechar-

acteristicsoftheplantstodeterminethedifferencesbetween

KL908andNJ6,andamongtheF

2

individualswithrespecttotheir

culmlengths,tillernumbersandothercharacters.

ResponseofKL908toGA

Elongationofshootswasquantifiedbyusingthemicrodropmethod

ormationofthenewSSRmarkers

MarkersPrimersequencesThesizeofthePCRproduct(bp)Repeatsequence

Ds3F:5\'-GGGTCTAATCAGATCTATGGTT-3\'190(ct)

17

R:5\'-TGTCTAGCTAACTGTGGTTGA-3\'

K81114F:5\'-GCTTCATCTTTGGCGACCACCGT-3\'126(tc)

21

R:5\'-TCCACCGGATTCACGAGATAA-3\'

GeneticMappingofRiceDwarfGeneD-53451

describedby自是花中第一流写的是什么花 Murakami(1968),ds

ofricevarietiesKL908andNJ6weresurface-sterilizedwith1.

5%NaClOfor30min,washedwithsteriledistilledwaterthree

minatedseeds

wereplacedona1%agarplateandgrownunderfluorescent

lightat30pproximately2d,1LofasolutionofGA

3

(3

10–2mol/L)inethanolwasappliedtothecoleoptilesofrice

r3dlaterthelengthsof

thesecond-leafsheathsweremeasured.

Forassayof-amylas蒙古包的拼音 eactivity(LanahanandHo1988),rice

ultingembryolesshalf-

seedsweresurface-sterilized,washedwithsterilizeddistilled

water,andplacedonastarchplatecontaining0.2%starch,2%姓名藏头诗在线制作

agar,10mmol/Lsodiumacetateand2mmol/LCaCl

2

GAplatesweremadebyadditionof1mol/LGA

3

tothecool

medium,andplateswithoutGA

3

he

plateswereincubatedinthedarkfor4dat30C,theywereput

intoiodinevaporfordetectingwhetherthestarchhadbeendi-

gestedby-amylase.

GeneticmappingoftheD-53gene

InadditiontothereportedSSRmarkers(McCouchetal.2002),we

searchedforotherSSRmarkersonchromosome11andother

hatwerepolymorphicbetweenthetwo

parentswereselectedasmarkersforgeneticmappingofD-53.

RecessiveplantsoftheF

2

populationfromthecrossbetween

KL908andNJ6wereusedtoanalyzethelinkagerelationship

o-

typesoftheindividualswerecategorizedandanalyzedwith

Mapmaker/Exp3.0softwaretoconstructalinkagemap(Lincolnet

al.1992).

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