梅花品种资源叶绿体基因组SSR研究
p29 9 lt;12007 M1 nbsp; lt; v JOURNALOFBEIJINGFORESTRYUNIVERSITYVol.29, Supp.1Jan., 2007l 2006-11-16htp [ “SE1 S39;[ “300070641、SE lt; S/Z39;[ “2005004。BT g ,p V, q。1Z_ 39;。010-82376017-601 EmailL 100083 lt;v10|Q。 39; s8yFSSR吕英民1, 2 殷 婧1 杨 果1 张启翔1,21 lt;v 2SE/ K1 B 1 s8DNAcpDNASSRNTCP9 948z yFDNA, s8DNASSRS k。[48 cpSSR nbsp;,NTSYSpc2.11 q9Nei39;s1972L. “ 、e “ SM、Jaccard “ Dice “ ,isY 、 s。Vs cpDNAs U W“d1“。1oM ,cpSSR,m L Ying-min1, 2;YINJing1;YANGGuo1;ZHANGQi-xiang1, 2.cpSSRanalysisofPrunusmumeculti-varresources.JournalofBeijingForestryUniversity200729 Supp.148-53[Ch, 22 ref.]1 ColegeofLandscapeArchitecture, BeijingForestryUniversity, 100083, P, R.China;2 ChinaNationalCenterforFlowerEngineeringTechnique, Beijing, 100083, P, R.China.Forty-eightcpDNAfingerprintsofPrunusmumecultivarswerebuiltupwiththeprimerNTCP9 fromtobaccocpDNAbyusingthecpSSRtechnique.BasedoncpSSRfingerprintsdataof48accessionsofP.mumecultivars, thecomplexityofrelationshipandoriginationofP.mumecultivarswereanalyzedbyusingNTSYSpc2.11toclusterandarangementanalysisofNei39;s1972geneticdistance, SMcoefi-cient, JaccardcoeficientandDicecoeficient.Keywords Prunusmume, cpSSR, DNAfingerprint s8DNAchloroplastDNA, cpDNAM nbsp;o,yN PrunusmumecpDNA cpDNA ]。 Pinus、 ZeamaysZ [ 1-15] , ,n1cpDNASSRs。5cpDNASSR DNASSR lt;[ 16] , V_] 、、 cpDNAslM, uY T1“yZ ]。39; B 1Nicoti-anatabacumcpDNASSR 948z yFDNA,cpDNASSRS k、。Vs cpDNAs U W“d1“,[F lt; 39;、 ,[ 39; s0S ,By L. m 、s0S、1y X HTB I LN。1 材料与方法1.1 nbsp; k cpSSR k“ nV1 1 2 q 48 。1.2 nbsp;ksZE1.2.1 试验仪器及试剂39; k PN S jlt; S v L i4。h1 p 3K 、Z 3 3/ K 。 M EcoRI MseI、T4 1 g 3/ 。Taq 1 3 、S 3/ K3 。 nbsp;A nbsp;TAS 3/ K3 。9 lt;1 g 39; s8yFSSR49 k4SsB。V1 nbsp;cpSSR k“ VTABLE1 SerialnumberofP.mumesamplesforcpSSRI| I| I| 1 l39; 17 e39; 33 lt;39;2 m 39; 18 Be39; 34 xlt;39;3 v } sl39; 19 39; 35 sllt;39;4 sl39; 20 *39; 36 5lt;39;5 * sl39; 21 v39; 37 ;lt;39;6 K39; 22 B 39; 38 lt;39;7 K39; 23 v 39; 39 C lt;39;8 }39; 24 39; 40 39;9 A39; 25 2*39; 41 39;10 *x39; 26 FKl39; 42 39;11 v39; 27 U39; 43 39;12 39; 28 x39; 44 1lt;39;13 2 39; 29 39; 45 K 39;14 T39; 30 39; 46 lt;39;15 xlt;39; 31 }39; 47 3lt;39;16 lt;39; 32 739; 48 l39;1.2.2 引物筛选39; k13 、、 s8DNAcpSSR[ 3, 17-19] , 9 |5z yFDNAs , T8_ ,]61.54,K 9KzB 1 s8DNASSRNTCP9 yFDNA s8DNASSRS k。NTCP9 5′-CTTCCAAGCTAACGATGC-3′5′-CTGTCCTATCCATTAGACAATG-3′1.2.3 试验方法及试验数据的记录分析方法F.JamesRohlf2001NTSYSpc2.11 q9 M “ L. “ 。L. “ dijNei39;s1972 “ 。 “ SM“ 39;e “ SSMad abcdJ“ Jaccard“ SJa abcDice“ SC2a 2abcT,aV U OTUI “1” ;bV UBOTU“1”, 6BOTU“0”;cV UBOTU“0”, 6BOTU“1”;dV U OTU“0” 。[SAHNClusteringdF EUPGMA s,9 M “ r,iMX-COMPMatrixcomparisonCOPHprogram2Manteltest_,9 “ [ 20] 39; TM1“ rij。2 结果与分析2.1 nbsp; cpSSRm s s8DNAsV U W T1“。 P 915HSSR {, c /。m1 NTCP9mFIGURE1 cpSSRfingerprintofprimercombinationsNTCP92.2 nbsp; cpDNASSRM cpSSRsNTCP9m nm1 cT s, m。1Nei39;s1972L. “ m2 U。2e “ SM T m3 U。3Jaccard “ m4 U。4Dice “ m5 U。51 4 sT m6 U。50 nbsp; lt; v 29 m2 Nei39;s1972L. “ FIGURE2 cpSSRdendrogramofNei39;s1972geneticdistancecoeficientsm3 SM “ FIGURE3 DendrogramofsimplematchingcoeficientsSMm4 Jaccard “ mFIGURE4 DendrogramofJaccardcoeficientsm5 Dice “ mFIGURE5 DendrogramofDicecoeficients9 lt;1 g 39; s8yFSSR51 m6 nbsp;T1 mFIGURE6 MXCOMPmatrixcomparisonofdendrograms52 nbsp; lt; v 29 2.3 nbsp; cpDNASSR s2.3.1 相关系数 2维比较9 4 M1“ V[ A “ z; “ “ TM “ Dice d,M1“ rij-0.984 44;J M1“ rij-0.951 74;SM M1“ rijK,9r -0.923 36,t“ rij 39;0.9,39;rdzY。2-wayManteltestTV T zQ ccpSSR39; f , V[ AM1“ I TdM 。2.3.2 树状图聚类情况scpSSRNTCP9_48 “ cpDNA 14, 13 ,] _ 92.86。sYNei39;s1972L. “ 、Dice “ 、Jaccard“ SM “ T1 , V[C “ M],Nei39;s“ 、D“ 、J“ M] T,SM“ TM , {],v|48z ks BFBFv } sl39;。Fe39;、Be39;、39;、B 39;、v39; 、*39;、v 39;、 sl39;、K39;、 A39;、K39;、 }39;,1 、 。 F7 39;、 2*39;、 }39;、l39;、m 39;、39;、 39;、x39;、*x39;、 2 39;、 T39;、39;、39;、 K 39;、 39;、 39;,1、x 。 FFKl39;、 U39;、lt;39;、 lt;39;、lt;39;、xlt;39;、xlt;39;、lt;39;、 sllt;39;、 C lt;39;、;lt;39;、5lt;39;、 3lt;39;,1lt; 。F 39;、* sl39;、v39;、39;、l39;。 BF 1lt;39;。AFLP TM1 nbsp;VC;M],9]。K A]cpSSRScpDNAL.M AAFLPSDNAL.M , M “ r1。iV t s8y M],7 s 9Hq/, s8yFM ,9 ] ,MWs9 sHq/kY。 y V s8yF ],cpDNAL. p。3 遗传相似系数分析与讨论 nbsp;48z cpSSR {9 L.M “ ,T V, k48 WL.M “ M0.400~ 1.000, L.M“ 0.806。 , WL.M “ M0.533~1.000, L.M “ 0.824; sl WL.M “M0.533~0.667, L.M “ 0.600; WL.M “ M0.800~1.000, L.M “ 0.889;x WL.M “ M0.867~1.000, L.M “ 0.933; WL.M “ M0.733~1.000, L.M “ 0.880; WL.M “ M0.667~ 1.000, L.M“ 0.773;lt; WL.M “M0.733~1.000, L.M “ 0.926; WL.M “ M0.800 ~ 1.000, L.M “ 0.907。 1lt;39;、v } sl39; s8DNA vs,V Ms *, S 。 M “ MKv,V *, N 。 nbsp;48 cpDNASSRs, V[ A W iBcpDNAMs, L.1“ , s8DNA nbsp;o,Q Wy N。NTCP9_ cpDNAL.M r 1.000, A39;、v 39;、v 39;B 39;, V LL.M , s8DNA s/ z_。 VnYVBs cpSSR W T1“ 。[ TsV , WcpDNA L.M ,9iBL.s9 lt;1 g 39; s8yFSSR53 s。 T39;Cs “d, iB f , L.1 ,w 、 V, L.] H,BL.Ms, s8DNA TL., V U W* Bt T1“。 6,cpSSRkY AFLP, V4 s8y TL., T1“4 I。 Y 2 3 B , 2 jlt;v\\ q,q S i 、fo k “ “ H f。 nbsp;I D[ 1] JURGEF.Geneticflexibilityofplantchloroplast[J].Nature,1999, 398115-116.[ 2] WOLFEKH, LIWH,SHARPPM.Ratesofnucleotidesubstitu-tionvarygreatlyamongplantmitochondrial, chloroplast, andnu-clearDNAs[J].ProcNatlAcadSciUSA, 1987, 84249 054-9 058.[ 3] CHENGYJ, GUOWW, DENGXX.cpSSRAnewtooltoan-alyzechlorplastgenomeofcitrussomatichybrids[J].ActaBotani-caSinica, 2003, 458906-909.[ 4] PLOMIONC, LEPROVOSTG, POTD, etal.Polencontamina-tioninamaritimepinepolycrossseedorchardandcertificationofimprovedseedsusingchloroplastmicrosatelites[J].CanadianJournalofForestResearch, 2001, 31101 815-1 816.[ 5] VIARDF, EL-KASSBYYA, RITLANDK.DiversityandgeneticstructureinpopulationsofPseudotsugamenziesiPinaceaeatchloroplastmicrosateliteloci[J].Genome, 2001, 443336-344.[ 6] FERRISFCR,KINGRA,HEWITTGM,etal.ChloroplastDNArecognizesthreerefugialsourcesofEuropeanoaksandsuggestsin-dependenteasternandwesternimmigrationstoKnland[J].Hered-ity, 1998, 805584-593.[ 7] ISODAK, SHIRAISHIS, WATANABES, etal.Molecularevi-denceofnaturalhybridizationbetween, AbiesveitdhiandA.homolepisPinaceaerevealedbychlorplast, mitochondrialandnuclearDNAmarkers[J].MolEcol, 2000, 9121 965-1 974.[ 8] JASONWL,MICHAELJH, SCOTTAJ, etal.Cytogenomicanalysesrevealthestructuralplasticityofthechloroplastgenomeinhigherplants[J].PlantCel, 2001, 13245-254.[ 9] KATOS,YAMAGACHH, SHIMAOTOY, etal.ThechloroplastgenomesofazukibeananditscloserelativesAdeletionmutationfoundinweedazukibean[J].Hereditas, 2000, 132143-48.[10] ROSAA, FRANCOISL, MARIAT, etal.Chloroplastmicrosatel-litepolymorphismsinVitisspecies.[J].Genome, 2002, 4561 142-1 149.[11] RICHARDGO,JEFFREYDP.ChloroplastDNAsystematicsAreviewofsanddataanalysis[J].AmerJBot, 1994, 811 205-1 224.[12] SCOTTNS, POSSINGHAMJV.ChangesinchloroplastDNAlev-elsduringgrowthofspinachleaves[J].JExpBot, 1983,341 756-1 767.[13]1, T,. 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