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/Z39;[ “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/ ŒK1  B Ÿ1 s8DNAcpDNASSR„NTCP9 ”948z ‹ yFDNA,› s8DNASSRS k。[ƒ48  cpSSR ” nbsp;,NTSYSpc2.11 q9Nei39;s1972L.  “ ”、e† “ ”SM、Jaccard “ ”„Dice “ ”,isY› 、 †s。V˜s ‚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;oŸ,yN PrunusmumecpDNA  cpDNA ]Ÿ。 –   ‹Pinus、 ZeamaysƒZ [ 1-15] ,Œ  ,n1cpDNASSRs。5cpDNASSR Ÿ‚ ‚DNASSR Ÿ lt;[ 16] ,Œ V_ž‚] ‹、、 cpDNAs„lM, uY T1““y˜Z   ]。39; B Ÿ1Nicoti-anatabacumcpDNASSR„ ”948z ‹ yFDNA,›cpDNASSRS k、。V˜s ‚cpDNAs U  W“d1“,[F lt; 39;、 „ ,[ 39; s0S ‚,B„y L. m 、s0S、1y X HTB ‚• I„ L‹N。1 材料与方法1.1 nbsp;  k‹ cpSSR k“ nV1 Ÿ1 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/ Œ›K 。 M EcoRI MseI、T4  1 g‡ 3/ Œ 。Taq 1 3 、S 3/ ŒK3  。 nbsp;Aš nbsp;T„AšS 3/ ŒK3  。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 †žK39; 22 B 39; 38 •lt;39;7 K39; 23 †v 39; 39 C lt;39;8 }39; 24  39; 40 Ž39;9 A39; 25 2*39; 41 39;10 *x39; 26 FKl39; 42  39;11 v39; 27 U39; 43   39;12  39; 28 Žx39; 44 1›lt;39;13  2 39; 29 39; 45 K 39;14  T39; 30  ˆ39; 46  lt;39;15 †xlt;39; 31 }39; 47  3lt;39;16 Šlt;39; 32 739; 48 l39;1.2.2 引物筛选39; k13 Ÿ 、、 Ž s8DNAcpSSR„[ 3, 17-19] , ”9  |5z ‹ yFDNA›s„ , T8_ž  Ÿ,]61.54,K ”9KzB Ÿ1 s8DNASSR„NTCP9 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 Ÿ“ ”L.  “ ”。L.  “ ”dijNei39;s1972  “ ”。 “ ”SM“ ”39;e† “ ”SSMad abcdJ“ ”Jaccard“ ”SJa abcDice“ ”SC2a 2abcT,aV U OTUŸI ’“1” ”;bV UBOTU“1”, 6BOTU“0”Ÿ”;cV UBOTU“0”, 6BOTU“1”Ÿ”;dV U OTU“0”Ÿ ”。[SAHNClustering›dF ƒ Œ EUPGMA s,9 M Ÿ“ ”r,iMX-COMPMatrixcomparisonCOPHprogram›2Manteltest_,9 ˜†“ ”[ 20] 39; T˜†M1“ ”rij。2 结果与分析2.1 nbsp; cpSSRm  s s8DNA’sV˜ U W T1“。  P„ ”915HSSR Ÿ{, c /。m1 „NTCP9mFIGURE1 cpSSRfingerprintofprimercombinationsNTCP92.2 nbsp; cpDNASSRM Ÿ cpSSRs„NTCP9žm nm1 cT› s,ž Žm。1Nei39;s1972L.  “ ”ž Ž m2 U。2e† “ ”SMž T m3 U。3Jaccard “ ”ž Ž m4 U。4Dice “ ”ž Ž m5 U。51 4 sT m6 U。50   nbsp; lt; v   29 m2 Nei39;s1972L.  “ ”ž ŽFIGURE2 cpSSRdendrogramofNei39;s1972geneticdistancecoeficientsm3 SM “ ” ŽFIGURE3 DendrogramofsimplematchingcoeficientsSMm4 Jaccard “ ”ž mFIGURE4 DendrogramofJaccardcoeficientsm5 Dice “ ”ž mFIGURE5 DendrogramofDicecoeficients9 lt;1 g‡   39; s8yFSSR51 m6 nbsp;T˜†1 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 “ ” ” Ž˜† z;  “ ”ž“ ” ”„ ŽTM Ÿ“ ”Dice ”„ Ž˜†d,˜†M1“ ”rij-0.984 44;Jž ”„ Ž˜†M1“ ”rij-0.951 74;SMž ”„ Ž˜†M1“ ”rijK,9rž -0.923 36,ƒt˜†“ ”rij 39;0.9,39;˜†rždzY。2-wayManteltestTV T “zQ˜ ccpSSR39; f ƒ, V[ AžM1“ ” I TdM 。2.3.2 树状图聚类情况scpSSR„NTCP9_48  “ cpDNA ”14, 13  Ÿ,] _  92.86。sYNei39;s1972L.  “ ”、Dice “ ”、Jaccard“ ”„SM “ ”ž T›1 , V[C –“ ”‚M],†Nei39;s“ ”、D“ ”、J“ ”žM] T,„SM“ ”ž TM ,  ” {‚],v|48z  k‹s BFBFv } sl39;。Fe39;、Be39;、39;、B 39;、v39; 、†*39;、†v 39;、‘ sl39;、†žK39;、 A39;、K39;、 }39;,1 ˜、 ˜ 。 F7 39;、 2*39;、 }39;、l39;、m 39;、39;、 ˆ39;、Žx39;、*x39;、 2 39;、 T39;、39;、Ž39;、 K 39;、 39;、  39;,1˜、x˜„ž ˆ 。 FFKl39;、 U39;、Šlt;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 1›lt;39;。AFLP TM1 nbsp;VC;M],9‚]。K A‚]cpSSRScpDNAL.M Ÿ AAFLPS‚DNAL.M Ÿ,‚  M Ÿ“ ”rž1。ƒi‚V ƒt   s8y› †M],7 s„ ”9Hq/, s8yFM Ÿ ,9 ]Ÿ ,MWs9  s„Hq/kY。 y V  s8yF    ]Ÿ,˜†cpDNAL. p。3 遗传相似系数分析与讨论 nbsp;48z ‹ cpSSR Ÿ{9 L.M “ ”,T V, k48  WL.M Ÿ“ ”M–0.400～ 1.000, L.MŸ“ ”0.806。 , ˜  WL.M Ÿ“ ”M–0.533～1.000, L.M Ÿ“ ”0.824; sl˜  WL.M Ÿ“”M–0.533～0.667, L.M Ÿ“ ”0.600; ˜  WL.M Ÿ“ ”M–0.800～1.000, L.M Ÿ“ ”0.889;x˜  WL.M Ÿ“ ”M–0.867～1.000, L.M Ÿ“ ”0.933;˜ WL.M Ÿ“ ”M–0.733～1.000, L.M Ÿ“ ”0.880; ˜  WL.M Ÿ“ ”M–0.667～ 1.000, L.MŸ“ ”0.773;lt; WL.M Ÿ“”M–0.733～1.000, L.M Ÿ“ ”0.926;ž ˆ  WL.M Ÿ“ ”M–0.800 ～ 1.000, L.M Ÿ“ ”0.907。 1›lt;39;、v } sl39; s8DNA   “vs,V ƒ  Ms *,   S 。 ˜  M Ÿ“ ”M–Kv,V ƒ ˜    *, N 。 nbsp;48  cpDNASSRs, V[ Až  W –iBcpDNAMs,Œ L.1“ –“,ƒ  s8DNA nbsp;oŸ„Ÿ,Q˜   WyŒ N。„NTCP9_ž  cpDNAL.M Ÿrž 1.000, A39;、†v 39;、v 39;B 39;, V ’ LL.M ,  s8DNA s„/‚ “z_ž。 VnYVBs„ Ÿ›cpSSR W T1“ ‚。[ TsV ,  WcpDNA L.M Ÿ,Œ9iBL.s9 lt;1 g‡   39; s8yFSSR53 „s。  T39;˜†Cs “d,Œ –iB
 f ƒ,  L.1 ,w 、 V,  †ŸL.Ÿ] H,BL.Ms,Œ s8DNA † TL., V U W* Bt T1“。 6“,cpSSRkY ‚ AFLP,Œ 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,.  SSRs„[J].q, 2003, 215444-448.[14] ,f ,, .SSRs0S 7/ ŒZ[J]. 2 jlt;, 2002, 154106-109.[15] W, gZ,,. s8DNA  RFLPsB“d‹[J].s , 2000, 38297-110.[16]u, nbsp; 3, ›p. „k •8 s8’s[J]. lt; S, 2000, 36346-51.[17]f, F, . 39; ‹ •8 s8SSRs[J].L., 2004, 264486-490.[18] 7. SSRk ŸŒ0B[J].0,2003123-25.[19] { , d b, .. nbsp;2l š、Ÿ7 3l š Yl š s8DNAKŸ M m [J].L., 1995, 1764-6.[20] X. 3 ”[M]. S, 1999.[21] ..S  m[M].S lt;, 1989.[22] .s0S ZE[M].  2 S/ Œ, 2003.责任编辑 李 慧/p