全 文 ：广西植物Guihaia29(5)：569—575 2009年9月
苏铁属植物RAPD反应体系的优化
及部分种类亲缘关系的探索
黄玉源1，5*，农保选2，3，刘驰2，钟晓青4，5，韦莉萍6
(1．仲恺农业工程学院生命科学学院，广州510225，2．广西农业科学院水稻研究所，南宁530007；3．广西作物
遗传改良生物技术重点开放实验室，南宁530007；4．中山大学生命科学学院，广州510275，5．中山大学
园林及生态经济规划设计研究所，广州510275I6．广西农业科学院科技情报研究所，南宁530007)
摘要：通过优化苏铁属植物的RAPD反应体系，进而探讨苏铁属部分种类的亲缘关系。结果表明，M92+、
dNTP、Taq酶及随机引物浓度在RAPD反应中有重要影响，而模板DNA浓度有一个很大的适应范围。适合苏
铁属植物RAPD分析的反应体系：20pL反应体系中，含有i0mmol／LTris-HCl(pH8．3)、50mmol／LKCI、2．0
mmol／LMgClz、200／_tmol／LdNTP、0．3t比mol／L随机引物、模板DNA50ng、Taq酶1．0U。聚类分析结果基本反
应了苏铁属各个种间的亲缘关系，证明了RAPD适用于苏铁属种间亲缘关系分析。RAPD聚类分析结合形态学
研究表明：海南苏铁、台湾苏铁、广东苏铁、滇南苏铁和仙湖苏铁之间的亲缘关系较远，支持各自成为独立的种。
关键词：苏铁屑fRAPD；亲缘关系
中图分类号：Q949文献标识码：A 文章编号；1000一3142(2009)05—0569一07
ImprovingofRAPDreactionsysteminCycasnd
studiesonthegeneticrelationshipsof omespecies
HI7ANGYu-Yuanl，5*，NONGBao-Xuan2，3，LIUChi2，
ZHONGXiao-Qin94，5，WEILi-Pin96
(1．CollegeofLifeSciences，ZD增地iUniversityofAgricultureandEngineering，Q埔ng曲ou510225，China)2．Rice
ResearchInstitute，Cnmng,．viAcademyofAgriculturalSciences，Nanning530007，China)3．GuangxiCropenetic
mprovementlandBiotechnologyLab。GuangxiAcademyofAgriculturalSciences，Nanrfa培530007，China)4．&hod
ofLifeSciences，SunYat-smUniversity，吼砒唱z110u510275，China；5．LayoutandDesign．Institute．ofLandscape
andEco-economy，SunYat-senUniversity，Guangzhou510275，China1 6．AgriculturalScientificandTechnology
InformationResearchInstitute，GuangxiAc demyofAgricultural＆妇，麟，Nanning530007，China)
Abstract：Inthispaper，thereactionsystemofRAPDinc3憾shadbeenestablishedandappliedtodiscusstherela-
fionshipsamongc．hainanensis，c．taiumniana，Cguangdo gensisC．diannanemisndC．fairy／akea．Theresults
showedthatRAPDwasaffectedbytheconcentrationsoffourcomponentsi-己M92+，dNTP，TaqDNApoIym盯8∞
andprimers．Theoptimalreactionsystemwas拍follows20弘Lreactionvolume，including10mmol／LTris—HCI(pH
8．3)，50mmol／LKCI，2．0mmol／LMgCl2，200／1moi／LdNTP，0．3ttmol／Lprimers，50ngtemplateDNA，1．0U
TaqDNApolymerase．ThereflectionofRAPDclusterothegeneticrelationshipofallthespeciesshowedthat
RAPDwassuitableforanalysisof nterspedfiegeneticrelationshipi Cycas．GeneticrelationshipanalysisbyRAPD
markersshowedthatfivedefinedspecieshadfargeneticrelationshipseachother，combiningwitht eanalysisofmot-
phologicaltraits，it’sconcludedthattheywerefiveindependentgeneticunitsat pecieslevel．
Receiveddate：2008—03-20Accepteddate：2009-01—15
Foundationitem：SupportedbytheNationalN turalScienceFoundationofCIlim(30260007)
BiographyzHUANGYu-Yuan(1959-)-male，borninQinzhouofG angxi，professor，Ph．D．，mainlyengageinstudiesofplantsysterrmties，
evolutionandecology．
。Authorfocorrespondence，E-mail,lqxhyy@yahoo．∞m．cn，hyy233@to札corn
万方数据
570 广西植物 29卷
Keywords：Cyc口5,RAPD；geneticrelationships
Lotsofstudiesofcycadswhichisknownas“living
fossil”forhumanbeing，havebeencarriedoutodiscuss
theevolutionaryrelationshipsamongdifferentcycads
species，originandmmigrationofexistingeyeads，andits
mechanismadaptedtovariousenvironments(Husng，
2001)．However，theclassifieationofsomespeciesiSstill
obscure，andthereiSfldisputeonwhetherthetaxaof
somespecieswerecredibleornot(Huang，2001)，espe—
dallyontheCycashainanensis，Ctaiwaniana，C
guangdongensisor“Guangdongeyead”(10calname)，C
diannanensis。Cfa rylakeaandSOOILIthasbecomea
barrierfortheutilizationndconservationofCyca5．
StudieshadshowedthatRAPDwasuitableforthea—
nalysisofimerspecificgeneticrelationship(Lanner甜
a1．，1996；Zhou，1999；Wang甜a1．，2003；Ⅸng甜盘Z．，
2005lXiao，2006；Cheneta1．，2007；Wang&Ding，2007，
Wangetal．，2008；Zhang&Huang，2008)．However，no
studyonCycasbyRAPDmarkerwaseportedinformer
researchesInthisresearch。wetryto stablishanopti—
mizedreactionsystemofRAPDinCycasndtodear
someproblemsofspeciesatmolecularlevel，thereby，
providereferencefors ttingupthemolecularbiology
basisinevolvementinvestigation，andprovideth ories
basisforfurtherstudyonthetaxonomy，propagation，u-
tilizafionandconservationofcycads．
1 Materialsandmethods
1．1Experimentalnmterials
sixspeaesofCycashadbeencollected(nble1)，7
--15samplesforeach．TheleavesofCycasweredried
withsilicagelrightafterbeingcollectedfromtheplant
(Chase&Hills，1991)．ThesampleswhichDNA
couldn’tbeextractedimmediatelywerestoredat-70℃
refrigerator．
InordertOascertainoptimumreactionsystem，cy—
casbalamaeQWarburgwasusedasthexperimental
materialtocarryOUthecomparatives udy．Otherfiv
definedspeciesw recarriedoutforthemolecularevolu—
tionaryrelationshipsresearch,
Table1 Colonyanditsoriginofsixspecies
GXAC,HerbariumoftheAgriculturalCollegeofGuang】dUniveraity．
1．2DNAextraction
Everyindividualofspecieswasmixedinequal
weightrespectively，thentheDNAoftheseamples
wereextractedbyusingCTABmethod(DoyleJJ＆
DoyleJL，1987)．
1．3P崃reaction
121piecesofprimerswerepurchasedfromShag-
halSangonBiologicalEngineeringTechnology&Services
CompanyLtd．，China．AnddNTP，TaqDNasew re
boughtfromTaKaRaBiotechnology(Dalian)Company
Ltd，Dalianof跚I】a．
Aseriesofconcentrationsandv lumesof achf赫
torweredesignedasfollows：DNA(10，25，50，75。i00ng
DNAineverytube，respectively)；M酽十(o．5，1．0，1．5，
万方数据
5期 黄玉源等：苏铁属植物RAPD反应体系的优化及部分种类亲缘关系的探索 571
2．O，2．5，3．0mmol／L)；dNTP(50，100，150，200，250，300
mnol／L)；TaqDNese(0．2，0．6，0．8，1．0，1．4U)．The
primerconcentrati‘onswere0．1，O．2，0．3，0．4，0．5，1．0，
2．0mnol／LThereactionwasconductedinBiometm
TgradientpCRapparatus．TheoptirmlPCRamplifica-
tionprocedurewasasfollows：Pre-denaturadonat94℃
for5minand35cyclesofdenaturationat94℃for30S。
annealingat37℃for40s。lengtheningat72℃forIi0
s；thenlengtheningat72℃for7minagai地Thefinal
productwaskepta 4℃．TheproductionofPCRwasi—
sohtedbyeleetrophoresisonanagarosegel(1％)and
dyedwitIlethidiumbromide(髓)，thenwereobserved
andphotographedthroughtheImagineMaster臃
1．4Dataanalysis
Everyeactionwasrepeatedforthreetimes．It
waslabeledwith1 or0，respectively，accordingtO
whetherithadRAPDbandornot．Thedatawereused
togeneratelaccardsimilaritycoefficientandtocon-
structa dendrogramaccordingtounweightedpair-
grouparithmeticaverageclustering(UPGMA)inNT—
SYSprograms．
2 Resultsandanalysis
2．1RAPDreactionsystemofCycas
2．1．1Theffectof emplateDNAconcentrations
ThebandsshowedclearlywhenthetemplateDNA
rangedfrom25--75ng，butobscurelywhenDNAless
than10ngormotethan75ng(Fig．1)．It’Sindicated
thattherangeofDNAconcentrationsWaSn tseverely
affectedintheR桃GiventhesuitablerangeofDNA
concentrations。itc uldobtainstableamplifiedpro ue—
don．ButtOavoidtheinfluenceofexcessiveDNA．every
tube(20／*Lreactions lution)contained50 gDNAwas
better．
2．1．2meffectofM92+concentrationsMg+plays
animportantroleinthePCRreaction。Fig．2showedthe
results0ftreatmentswi hdifferent咐+concentra—
tions．‰theM酽+concentratiorLw s0．5mmol／L，
onlyonebandwasproduced,whenitwas3．0mmol／L，
thedarkenbackgroundWaserious；whenit asinthe
rangefrom1．0—2．5mmol／L。similarbandscouldbe
seen，andwhentheconcentrationwas2．0mmol／L，the
Fig．1Comparisonofamplificationeffectsftemplate
DNAwithdifferentco centrations．
Fromleftoright(Primer$404)110。25，50，75，100rigl
M：GeneRulerTMDNALadderMIX
Fig．2EffectsofM92+concentrationsonamplifiedresults
Fromleftofight(Primer$216)i3．0，2．5，2．O，1．5，
1．0，0．5mmol／L，MlDNAMarkerDL2000
Fig．3EffectsofdNTPconcentrationsonamplifiedresults
Fromldtofigh(PfimerS347)；50，100，150，200。
250，300／_‘mol／L,M：LambdaDNA／Hin Ⅲ．
bandswereclearest．Soweselectedthe2．0retool／Lof
Mg+asthebestconcentration。,．
2．1．3TheffectofdNTPconcentrationsTheampli—
fledproductionwerebasicallysimilarwhentheconcen-
trafionsofdNTPrangedfrom5Q一200弘mol／L(Hg．3)，
butit wasclearerwhentheconcentrationswas200
／比mol／L,whenitwas250t比mol／L，thebandscouldntbe
万方数据
572 广西植物 29卷
魄ndearlylwhenitwas300肛mol／L，therewash’tany showedtheamplifiedbandsofprimerS380．
hmdSoweselected200胆,nol／LdNTP髓thebest
contentforR啪resction
2．1．4neeffectofTaqDNApolymeraseFig．4in。
dicatedhatwhentheTaqDNApolymerasenged
from0．8—1．4U／pL，theamplifiedban swerealmost
consistentswhenitWaSlessthan0．6U／肛L，thebarld8
weregetting1TiDreandmoreobscure．Consideringthe
stabilityandcoSt，weselected1．0U／弘LTaqDNa8e
forRAPDreaction．
2．1．5TheffectofprimersconcentrationsThere-
searchresultsshowedthatwhentheprimerconcentra-
tionsrangedfrom0．2—0．5tanol／L,itcouldproduce
basicalsimilarb nds；whenitwas0．3t比mol／L(Fig．5)，
thebandswereparticularlydear，theprimerconcentra。
tionsWaSlessthan0．2／maol／Lormorethan0．5弘mol／
L，theamplifiedban sincreasedndgotobscure．Sowe
selected0．3／比mol／LprimerfoRAPDreaction．
Fig．4EffectsofTaqDNApolymerase
concentrationsonamplifiedresults
Fromleftoright(PrimerS24)11．4·1．O·0．8·0．6，
0．2U／止，MlGeneRulerTMDNAIadd盯MIX．
2．2Primersselection-DNAamplifiedandclustering
analysls
2．2．1Primersselection3 significantdifferentspe-
desinmorphologicwereamplifiedw th121primersto
screenpolymorphiemark rsforclusteringanalysis．21
piecesofprimerproducedstably，clearlyandpolymor-
phiebandswereselected(，】Xble2)andappliedtothe
furthersnalysisoffivespecies．1btal202DNAbands
hadbeenamplifiedrom21primerswithanaverageof
9．6bandsperprimer．whilet erw re170bandsof
thepolymorphieDNAoccupying84．16％oft tal,
showingtherew regreatdiversityinC casinterspe。
des。thebandrangedfrom200—3000bp．Fig．6
Fig．5Effectsofprimerconcentrationsonamplifiedresults
FromlefttOfight(Primer$216)lO．1，O．2,0．3-0．4，
0．5。1．O．2．Opmol／L,MlDNAMarkerDL2000．
Fig．6Amplifiedban sofprimer$380forfivespecies
1．Cfairylakea1 2．Chainancnsis,3．c．taizmniana,4．C
guan&dongenses；5．c．diannanensis(q-)·6．c．diammntmjis
(台)IM,GeneRulerTMDNAIMIdderMix．
Table2 21primersusedinanalysis
Sequence
(5-3’) Am。p。ldifiedPⅢoly㈣mor-Ppe留rcenta驴ge
of
loci phicIociPi=‘澎：“
S8 GTCCACACGG11 11 100．0
鸵2 TGCCGAGCTG10 10 100．0
S29GGGTAACGCC12 9 75．0
$31CAATCGCCGT11 9 81．8
S32TCGGCGATAG14 13 92．9
$36AGCCAGCGAA8 7 87．5
$38AGGTGACCGT11 7 63．6
S40GTTGCGATCC9 7 77．8
S41ACCGCGAAGG6 4 66．7
S102TCGGACGTGA7 6 85．7
S107CrGCAToGTG5 3 60．O
S132ACGGTACCAG10 9 90．O
S151GAGTcTCAGG10 9 90．O
S156GGTGACTGTG11 9 81．8
$198CTGGCGAACT7 7 100．0
$216GGTGAACGCT9 8 88．9
S380GTGTCGCGAG13 12 92．3
S404GGCGGTTGT9 8 88．9
S410TCTGGCGcAC7 4 57．1
S464GTGTCTCAGG1l 10 90．9
S1427GTGGCCGATG11 9 81．8
万方数据
5期 黄玉源等。苏铁属植物RAPD反应体系的优化及部分种类亲缘关系的探索 573
Itwaslabeled1or0，respectively．accordingto
whethertherewasRAPDbandornot．Thedata
wereanalyzedbyNTSYSsoftware，weadopted
Jaccardcoefficientandrewtheclusteringmapu—
singUPGMA(Fig．7)．
一口．～Irylmkee
2-0．hm|n●n●n●im
3。口taiwmni#nm
4。口Eumnldonlon#／e
5’0．dimnnmnen#，·(旱)
6一口．dlmnnaneneis(8)
Fig．7DendrogramgeneratedbyJaccard
coefficientofsimilarity
Table3 Similaritycoefficientandgenetic
distanceamong6taxa
2．2．2austering砌灿msimilaritycoefficient
betweenverytwospeciesrangedfrom0．3636一O．5597
(Table3)，showedthattheyhadafargeneticrelation-
shipeachother．Thebiggestsimilaritycoefficientwas
0．5597．whichlo atedinChainanensisa dc．taizmn／一
aMa，indicatingthattheyhadclosestgeneticrelationship．
T11esimilaritycoefficientofCfairyl嫩eaandC
guangdongenseswasthesmallestamongallspecies，it
showedthattherew refarthestgeneticrelationshipbe-
tweenthem．Thesimilaritycoeffidentofdifferents x
colonyina蛳spedes·c．diannanensiswas0．9098，
whicllwasthebiggestdatumofthe81lmaterialsfthey
clusteredfirstofaU，showingtheclosestgeneticrelation-
shipinallmaterialsTheresultsofRAPDdusterthat
reflectedthegeneticrdationshipofallthespecies
showedthatR栅wasverysuitableforinterspedesge—
neticrelationshipsnalysisofcycad．
3 Discussions
3．1Severs／factorsaf钕地喀theIl址毋rmtionand
RAPDm蛆tislitytoplanttaxonomy
TemplateDNA，M酽+，dNTP，砌DNApolymer-
啪andtheprimerswerethemainfactorsaffectinghe
RAPDreaction。ifoneo themwaschanged，thestability
ofwholereactionsystemcouldbeinfluencedgr ady．So
itisimportantoascertainhereactionparametersfor
RAPDanalysis．
TosuHlmarizeabover sultsandanalysis。theop-
tim．a／reactionsystemofRAJPDwasasfollowst20弘L
reactionv lumeincluding10mmol／LIris—HCl
(pH8．3)，50mmol／LKcl，2．0mmol／LMgCI=，200
／llmol／LdNTP，0．3vmol／Lprimers，50ngtemplate
DNA，1U屁gDNApolymerase．
Manyresultsofcomparatives udi sonRA玉，】Dwith
触methodstomanyfamiliesofplantsshowedthat
theresultswereveryhighcoherence．Suchascorrela-
lionanalysisamonggenetics milaritiesbas donRAPD·
ISsRmarkersandphenotypicdatashowedthecorrela-
doncoeffidentofRAPDand勰markswere0．932，
indicatingtheyweresignificantlyorrelated．Thesum-
betoffragmentsperprimer，the、percentage．ofpolym r-
phiclociwasinturn9．09％，83．73％forRAPDmark-
ra，and5．88％，79．79％forISsRmarkers．It’s
showedthattheseindexesofRAPDwerehigherthan
阻AlthoughISSRhadhigherpercentageofpoly—
morphiclocinCannuunlthanRAPD，andeffective
numberofalleles(Ne)，Shannon’sInformationindex(1)，
heterozygosity(Ht)，thegene icd ffer ntiationcoeffident
(Gst)basedonISsRmarkerswerebiggerthanthose
basedonRAPDaswell．Buttheseindexeshadvery
smalldifference．thenumb rcommonlywasthescopein
0．01--0．04，whereassomenumberswere砘廿Dhigher
thanIS；SR(Chen甜a1．，2007)．
Inanotherstudy，forRAPDanalysis，13random
primerswereidentifiedw thpotymorphismamongthe
entries；intotal，167bandswereproduced，ofwhich
50bandswerepolymorphic；percentageofpolymorphic
bandswas29．9％．ForISSRanalysis，7primerswere
万方数据
574 广西植物 29卷
identifiedw thpolymorphismamongthentries；into—
tal，96bandswereproduced，ofwhich44bandswere
polymorphic；percentageofpolymorphichandswas
45．8％(Xiao，2006)．
StudyresultsfromZhang砑a1．(2007)showed
that225and196bandswereamplifiedby12RAPD
primersand12ISSRprimersre pectively．Therewere
215and196polymorphicbandsamplifiedbyR址D
andISSRprimersre pectively，andthepercentagesof
polymorphiebandsw re95．56％and100．00％re—
spectivdy．Thegenediversityamongspecies(Ht)and
withinspedes(Hs)were0．3689，0．3575and0。1077，
0．1380，respectively．
Hereinbeforetheser sultsindicatedthatpolymor．
phicbandsamplified，percentagesofpolymorphicbands
generalwerehigherRAPDthanISSR．inthegenediver-
sityamongspecies(Ht)andwi hispecies(Hs)Ne，I，Ht
andGst，RAPDandISSRwereveryappropinquity，and
蛐specieshadhigherISSRthanRAPD，softiespecies
hadhigherRAPDthanISSR，buttheirnumberdiffer-
enl2ewanverysmall．
ThesetudiesfullyprovedthatRAPDmethodwas
averytrustyandstabilizationtoanalysisplantrelation-
shipbetweenspecies，subspedes，variationandform，even
variety(Wangeta1．，2003；Qieta1．，2003，2004；Dinget
a1．，2005；Xiao，2006；Chert功a1．，2007；Wang萏LDing，
2007；Xiaoet 1．，2007；zl捌唱eta1．，2007；Jia以a／．，
2008；Panet L，2008；Wang疗a1．，2008；Zhangeta1．，
2008)．Aftersomescholarshadcompletedtheompara—
tivestudiesRAPDandISSR。provedRAPDaveryand
exactmethodt researchtherelationshipsbetweenspe—
des，subspecies，variationandform，theycontinueto se
RAPDforthefartherstudieson omespeciesn8anle
genera，suchasZhangeta1．hadmadethestudieswhich
comparatives udi sRAPDand黜OllIrisLPLants
andpublished(2007)，thentheyconti uedtocarryout
RAPDstudiesonotherspeaesofMsLplants(2008)．
TheimprovingofRAPDreactionsystemforCycas
inthistudywasignificanttoaffordrightandeffective
methodonanalysedtherelationshipbetweenspecies，
subspecies，variationandforminCycadopsida．
3．2Evolutionaryrelationshipsof ve删簋
Researchershavebeendisputedonthetaxonf￡
fairylakea，Chainanensis。Ctaiwaniana，Cgu ngdon—
gensis．Cdiannanensisforalongtime．BothHuang
(2001)andW g以a1．(1996)consideredthatCycas
guangdongensis(thebrandhanginthe翻mpletreloca-
tedinShenzhenFaj巧LakeBotanicalGarden)andCtai-
zmn／anaweredifferentnalneso{asamespedes．Chen
pointedhatChainanensisdi tinguishedfromCtail-
n／anabymegasporophyllwithlesslateralsegments，and
843on(Hill，1994)．Wangetal．(1996)madewideinvesti-
gationinHainanProvince，findingthatMegasporophyU
terminalsegmentsshapeoftypeformofCtai纰niana
andChainanensiswerebroaddeltoid，furthermore，it
hadaseriesofvariationfromdeltoidtosubulate—lanceo-
late，numbersoflateregmentsalsohadacertainrange
ofvariation，soCAainanens／swasamodifiedform，and
shouldbemergedintothelatter．Huang(2001)consid-
eredthatcharacteristicsofChainanensishadbeenmis—
takenlyincludedinthemorphologicillustratingofCtai-
讹rdanabyWang，Wang(1996)thoughtCtaizmniana
hadmanyvarieties，becausehehadillustratedcharacter-
isticsofCtaiwanianaincludingallthemegasporophyll
shapesofChainanensis．Chcn(1999)mergedCdian-
nanensisandCfairflakeaintoC．taizoaniana，thenre-
namedCtaiwaniana’swhileCllinesenameasC
guangdongemis．Anatonficslresearchshowedthatmi—
cro-shapeswereapparentlydifferentb weenCtaitoan—
ianandChainanensis，theywerewoindependence
species(Huang，2001)．
Inthisresearch，differentsexesofCdiannanensis
wereclusteredinthefirstgroup，theirsim laritycoeffi-
dentwas0．9098．anditwasthebiggestoneofallthe
spies，Ctaizoaniana，Chainanensisa dCg猢枷一
gemiswereclusteredtogether，thesimilaritycoeffident
betweenachotherofthemrangedfrom0．5476——
0．5597．ItindicatedthatChainammiswasdoserge-
neticrelationshiptoGtaiwaniana，andChai anensis
andCguangdongensis，Ctaizmnianaandc．guang-
dongensisalsohadosergeneticrelationship．Thes mi-
laritycoefficientamongCfairylakea，Cdi nnanensis
andCtaiwJan／anar gedfrom0．3671一O．4395，andit
showedthattheyhadafargeneticrelationshipeachoth-
er．
AccordingtothecoiD．Iilonview(Lane,1993)，those
万方数据
5期 黄玉源等：苏铁属植物RAPD反应体系的优化及部分种类亲缘关系的探索 575
sp茂ieswhicht eirsimilaritycoefficientseachotherwA．q HillKD，ChenJR．1994．On觚tai姗danaCarruthars(Cyca—
molethan0．90couldbemergetothesalTlespecies，
thosewhichsimilaritycoefficientsra gedfrom0．67——
o．90weredifferentsubspeciesandvariations．thesimi—
laxitycoefficientsb inglessthan0．67wasatthespecies
leveLInpresentstudy，thesimilaritycoeffidentsof5
defmedspeciesofCycaswereall essthan0．67，there-
fore。theauthorssupportedthattheywereestablished嬲
fiveindependentspecies．
Analyfingtheresultsofmolecularmark，thesimi—
larityodeffidentwasveryhigh(morethan0．90)between
maleandfemaleofCdiannanensis。othersimilaritycod-
ficientofdefinedspecieseachotherwas10werandvery
low(allwerel ssthan0．687)，itwasapproximatelyco-
incidentwi ht eclassicaltaxonbyotherresearchmeth-
ods．ItindicatedthatR栅wasapropitiouswayto
studyC castaxonomy．Presentresearchisusefulfor
furtherr searchont evolutionaryrelationshipsofev-
crytaxonfCycasndaffordedeffectivelyusingmolecu-
tarmarkermethocL
Aeknowledgenmnts殇eprojectwacarriedoutatKey
LabofGuangxiCropGeneticImprovementandBio ech-
nologyandLabofCropGeneticandBreedingofAgricul-
turalCollege，GuangxiUniversity，Wethankthecrewin
thesetwolabsfortheirhelp．
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万方数据
苏铁属植物RAPD反应体系的优化及部分种类亲缘关系的探索
作者： 黄玉源， 农保选， 刘驰， 钟晓青， 韦莉萍， HUANG Yu-Yuan， NONG Bao-Xuan， LIU
Chi， ZHONG Xiao-Qing， WEI Li-Ping
作者单位： 黄玉源,HUANG Yu-Yuan(仲恺农业工程学院,生命科学学院,广州,510225;中山大学,园林及生
态经济规划设计研究所,广州,510275)， 农保选,NONG Bao-Xuan(广西农业科学院,水稻研究
所,南宁,530007;广西作物遗传改良生物技术重点开放实验室,南宁,530007)， 刘驰,LIU
Chi(广西农业科学院,水稻研究所,南宁,530007)， 钟晓青,ZHONG Xiao-Qing(中山大学,生
命科学学院,广州,510275;中山大学,园林及生态经济规划设计研究所,广州,510275)， 韦莉
萍,WEI Li-Ping(广西农业科学院,科技情报研究所,南宁,530007)
刊名： 广西植物
英文刊名： GUIHAIA
年，卷(期)： 2009,29(5)
被引用次数： 4次
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