梅花品种资源叶绿体基因组SSR研究

29 9 <12007 M1   < v JOURNALOFBEIJINGFORESTRYUNIVERSITYVol.29, Supp.1Jan., 2007l :2006-11-16htp: journal.bjfu.edu.cn “:SE1 S' “(300070641)、SE < S/?Z' “(2005004)。BT: g ,p V, q。1Z_: '。:010-82376017-601 Email:Luyingminbjfu.edu.cn :100083 <v10|Q。 '= s8yFSSR吕英民1, 2 殷 婧1 杨 果1 张启翔1,2(1 <v 2SE/ )K1: B 1= s8DNA(cpDNA)SSR(NTCP9) 948z yFDNA,= s8DNASSRS: k。48 cpSSR  $,NTSYSpc2.11 q9Nei's(1972)L. “ 、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.JournalofBeijingForestryUniversity(2007)29 (Supp.1)48-53Ch, 22 ref.1 ColegeofLandscapeArchitecture, BeijingForestryUniversity, 100083, P, R.China;2 ChinaNationalCenterforFlowerEngineeringTechnique, Beijing, 100083, P, R.China.Forty-eightcpDNAfingerprintsofPrunusmumecultivarswerebuiltupwiththeprimerNTCP9 fromtobaccocpDNAbyusingthecpSSRtechnique.BasedoncpSSRfingerprintsdataof48accessionsofP.mumecultivars, thecomplexityofrelationshipandoriginationofP.mumecultivarswereanalyzedbyusingNTSYSpc2.11toclusterandarangementanalysisofNei's(1972)geneticdistance, SMcoefi-cient, JaccardcoeficientandDicecoeficient.Keywords Prunusmume, cpSSR, DNAfingerprint= s8DNA(chloroplastDNA, cpDNA)M  o,yN (Prunusmume)cpDNA cpDNA 。 (Pinus)、 (Zeamays)Z 1-15 , ,n1cpDNASSRs。5cpDNASSR DNASSR < 16 , V_ 、 cpDNAslM, uY T1“yZ + 。' B 1(Nicoti-anatabacum)cpDNASSR 948z yFDNA,cpDNASSRS: k、。Vs) cpDNAs U W“d1“,F < '、 ,# ' s0S: $,By L. m 、s0S:、1y X HTB I L=N。1 材料与方法1.1   k cpSSR k“ (nV1) 1 2 q 48 。1.2  ksZE1.2.1 试验仪器及试剂' k PN (S j< S v L i4。#h1 p 3K 、Z 3 3/ K 。 = M EcoRI MseI、T4 1 g 3/ 。Taq 1 3 、S 3/ K3 。  A  TA:S 3/ K3 。9 <1 g : '= s8yFSSR49 k4:SsB。V1  cpSSR k“ :VTABLE1 SerialnumberofP.mumesamplesforcpSSRI| I| I| 1 l' 17 e' 33 <'2 m ' 18 Be' 34 x<'3 v sl' 19 ' 35 sl<'4 sl' 20 *' 36 5<'5 * sl' 21 v' 37 ;<'6 K' 22 B ' 38 <'7 K' 23 v ' 39 C <'8 ' 24 ' 40 '9 A+' 25 2*' 41 '10 *x' 26 FKl' 42 % ='11 v' 27 =U' 43 ='12 ' 28 x' 44 1<'13 2 ' 29 ' 45 K '14 $ T' 30 ' 46 <'15 x<' 31 ' 47 3<'16 <' 32 7' 48 l'1.2.2 引物筛选' k13 、 = s8DNAcpSSR 3, 17-19 , 9 |5z yFDNA+s , T8_ ,61.54%,K 9KzB 1= s8DNASSR(NTCP9) yFDNA= s8DNASSRS: k。NTCP9 :5-CTTCCAAGCTAACGATGC-35-CTGTCCTATCCATTAGACAATG-31.2.3 试验方法及试验数据的记录分析方法F.JamesRohlf(2001)NTSYSpc2.11 q9 M “ L. “ 。L. “ dijNei's(1972) “ 。 “ SM“ 'e “ :SSM=(a+d) (a+b+c+d)J“ (Jaccard“ ):SJ=a (a+b+c)Dice“ :SC=2a (2a+b+c)T,aV U OTUI “1” ;bV UBOTU“1”, 6BOTU“0”;cV UBOTU“0”, 6BOTU“1”;dV U OTU“0” 。SAHNClusteringdF (E(UPGMA) s,9 M “ r,iMX-COMPMatrixcomparisonCOPHprogram2Manteltest_,9 “ 20 (' TM1“ )rij。2 结果与分析2.1   cpSSRm # s= s8DNAsV U W T1“。 P 915HSSR ,: c /。m1 NTCP9mFIGURE1 cpSSRfingerprintofprimercombinationsNTCP92.2   cpDNASSRM cpSSR+sNTCP9m (nm1): cT s, m。1)Nei's(1972)L. “ m2 U。2)e “ (SM) T m3 U。3)Jaccard “ m4 U。4)Dice “ m5 U。5)1 4 sT m6 U。50   < v 29 m2 Nei's(1972)L. “ FIGURE2 cpSSRdendrogramofNei's(1972)geneticdistancecoeficientsm3 SM “ FIGURE3 Dendrogramofsimplematchingcoeficients(SM)m4 Jaccard “ mFIGURE4 DendrogramofJaccardcoeficientsm5 Dice “ mFIGURE5 DendrogramofDicecoeficients9 <1 g : '= s8yFSSR51 m6  T1 mFIGURE6 MXCOMPmatrixcomparisonofdendrograms52   < v 29 2.3   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 '0.9,'rdz)Y。2-wayManteltestTV T ?zQ: ccpSSR' f , V AM1“ I TdM 。2.3.2 树状图聚类情况+scpSSRNTCP9_48 “ cpDNA 14, 13 , _ 92.86%。sYNei's(1972)L. “ 、Dice “ 、Jaccard“ SM “ T1 , V?C “ M,Nei's“ 、D“ 、J“ M T,SM“ TM , +) ,v|48z ks BF:BF:v sl'。=F:e'、Be'、'、B '、v' 、*'、v '、 sl'、K'、 A+'、K'、 ',1 、 。 F:7 '、 2*'、 '、l'、m '、'、 '、x'、*x'、 2 '、 $ T'、'、'、 K '、% ='、 =',1、x 。 F:FKl'、 =U'、<'、 <'、<'、x<'、x<'、<'、 sl<'、 C <'、;<'、5<'、 3<',1< 。F: '、* sl'、v'、'、l'。 BF: 1<'。AFLP TM1  V?C;M),9)。K A)cpSSRS:cpDNAL.M AAFLPS:DNAL.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  48z cpSSR 9 L.M “ ,T V, k48 WL.M “ M0.400 1.000, (L.M“ 0.806。 , WL.M “ M0.5331.000, (L.M “ 0.824; sl WL.M “M0.5330.667, (L.M “ 0.600; WL.M “ M0.8001.000, (L.M “ 0.889;x WL.M “ M0.8671.000, (L.M “ 0.933; WL.M “ M0.7331.000, (L.M “ 0.880; WL.M “ M0.667 1.000, (L.M“ 0.773;< WL.M “M0.7331.000, (L.M “ 0.926; WL.M “ M0.800 1.000, (L.M “ 0.907。 1<'、v sl'= s8DNA vs,V Ms *, S 。 M “ MKv,V *, N 。  48 cpDNASSRs, V A W iBcpDNAMs, L.1“ , = s8DNA  o,Q Wy N+。NTCP9_ cpDNAL.M r 1.000, A+'、v '、v 'B ', V ? LL.M , = s8DNA s/ ?z_。 VnYVB+s cpSSR W T1“ 。 TsV , WcpDNA L.M ,9iBL.s9 <1 g : '= s8yFSSR53 s。 T'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 j<v q,q S i 、fo k “ “ H f。  I D 1 JURGEF.GeneticflexibilityofplantchloroplastJ.Nature,1999, 398:115-116. 2 WOLFEKH, LIWH,SHARPPM.Ratesofnucleotidesubstitu-tionvarygreatlyamongplantmitochondrial, chloroplast, andnu-clearDNAsJ.ProcNatlAcadSciUSA, 1987, 84(24):9 054-9 058. 3 CHENGYJ, GUOWW, DENGXX.cpSSR:Anewtooltoan-alyzechlorplastgenomeofcitrussomatichybridsJ.ActaBotani-caSinica, 2003, 45(8):906-909. 4 PLOMIONC, LEPROVOSTG, POTD, etal.Polencontamina-tioninamaritimepinepolycrossseedorchardandcertificationofimprovedseedsusingchloroplastmicrosatelitesJ.CanadianJournalofForestResearch, 2001, 31(10):1 815-1 816. 5 VIARDF, EL-KASSBYYA, RITLANDK.DiversityandgeneticstructureinpopulationsofPseudotsugamenziesi(Pinaceae)atchloroplastmicrosatelitelociJ.Genome, 2001, 44(3):336-344. 6 FERRISFCR,KINGRA,HEWITTGM,etal.ChloroplastDNArecognizesthreerefugialsourcesofEuropeanoaksandsuggestsin-dependenteasternandwesternimmigrationstoKnlandJ.Hered-ity, 1998, 80(5):584-593. 7 ISODAK, SHIRAISHIS, WATANABES, etal.Molecularevi-denceofnaturalhybridizationbetween, AbiesveitdhiandA.homolepis(Pinaceae)revealedbychlorplast, mitochondrialandnuclearDNAmarkersJ.MolEcol, 2000, 9(12):1 965-1 974. 8 JASONWL,MICHAELJH, SCOTTAJ, etal.CytogenomicanalysesrevealthestructuralplasticityofthechloroplastgenomeinhigherplantsJ.PlantCel, 2001, 13:245-254. 9 KATOS,YAMAGACHH, SHIMAOTOY, etal.Thechloroplastgenomesofazukibeananditscloserelatives:AdeletionmutationfoundinweedazukibeanJ.Hereditas, 2000, 132(1):43-48.10 ROSAA, FRANCOISL, MARIAT, etal.Chloroplastmicrosatel-litepolymorphismsinVitisspecies.J.Genome, 2002, 45(6):1 142-1 149.11 RICHARDGO,JEFFREYDP.ChloroplastDNAsystematics:AreviewofmethodsanddataanalysisJ.AmerJBot, 1994, 81:1 205-1 224.12 SCOTTNS, POSSINGHAMJV.ChangesinchloroplastDNAlev-elsduringgrowthofspinachleavesJ.JExpBot, 1983,34:1 756-1 767.131, T,. % SSRsJ.q, 2003, 21(5):444-448.14 ,f , .SSRs0S: 7?/ ZJ. 2 j<, 2002, 15(4):106-109.15 W, gZ,?,.= s8DNA RFLPsB“dJ.s , 2000, 38(2):97-110.16u,   3, p. !k 8= s8sJ. < S, 2000, 36(3):46-51.17f=, F, . ' = 8= s8SSRsJ.L., 2004, 26(4):486-490.18 7. SSRk 0BJ.0,2003(1):23-25.19 , d b, .:.  2l 、7 3l # Yl = s8DNAK = M m J.L., 1995, 17(6):4-6.20 X. 3 M.: S, 1999.21 .S mM.:S <, 1989.22 !.s0S: ZEM. : 2 S/ , 2003.(责任编辑 李 慧)