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,2016,36(6):1117-1124
犃犮狋犪犅狅狋.犅狅狉犲犪犾.犗犮犮犻犱犲狀狋.犛犻狀.
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犌犲狀犲狋犻犮犇犻狏犲狉狊犻狋狔犃狀犪犾狔狊犻狊狅犳犜犲犪犌犲狉犿狆犾犪狊犿犻狀犙犻犪狀狀犪狀
犘狉犲犳犲犮狋狌狉犲犫狔犛犛犚犕犪狉犽犲狉狊
ZHANGMingze,YAOYuxian,CHENShijun
(ColegeofBioscienceandAgronomy,QiannanNormalUniversityforNationalities,Duyun,Guizhou558000,China)
犃犫狊狋狉犪犮狋:Inthisstudy,fifteenSSRprimerpairswithpolymorphismwereusedtoassessgeneticdiversity
andrelationshipof60wildteagermplasmsfromQiannanprefecture.Theresultsshowedthatthepercent
ageofpolymorphicbands(犘犘犅)was98.64%.Atotalof147observedalelesand73.7786effectiveal
lelesweregenerated,withameanof9.8and4.9186perlocus.Totaly280genotypesweredetectedinal
materials,withameanof18.7foreachpolymorphismprimerpairs.Thepolymorphisminformationcon
tentvariedfrom0.1239to0.9268,withanaverageof0.5725.Theaverageobservedandexpectedhet
erozygositieswere0.4700and0.6023,respectively.TheaverageShannon’sinformationindexwas1.464
4.Thesimilaritycoefficientamong60teagermplasmswas0.2051to0.8636.Whenthesimilaritycoeffi
cientwas0.4775,eightmajorgroupsweregeneratedfromaltheaccessionstestedbyUPGMAclustering
analysis.Theresultsshowedthattherewerenosignificantcorrelationbetweengeneticandgeographicdis
tanceamongtheteagermplasms,andsomeindividualsinthesamepopulationdistributedindifferent
groupsofthecluster.Theseresultssuggestedthatthematerialsusedintheexperimentpossessedabroad
geneticvariation,showingahighlevelofgeneticpolymorphismamongteagermplasmsrevealedbySSR
markers.
犓犲狔狑狅狉犱狊:tea;germplasmresources;SSRmolecularmark;geneticdiversity;Qiannanprefecture
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Table1 Codeandoriginofteaaccessionsusedinthisstudy
ef
No.
¶^
Origin
ef
No.
¶^
Origin
ef
No.
¶^
Origin
Y1 cdgh Mochong,Duyun Y21 cdiBGujiang,Duyun W41 jc´kShuixiang,Shandu
Y2 cdBlJiangzhou,Duyun Y22 ÚmnmGoushan,Dushan W42 opqrZhucang,Wengan
Y3 ú@sYunwu,Guiding Y23 ú@sYunwu,Guiding W43 t+(Tongzhou,Pingtang
Y4 cdgh Mochong,Duyun Y24 cdiBGujiang,Duyun W44 t+(Tongzhou,Pingtang
Y5 cduv Huanghe,Duyun Y25 cdwxBaimang,Duyun W45 Úm1mYingshan,Dushan
Y6 cdymTuanshan,Duyun Y26 cdBlJiangzhou,Duyun W46 z{|} Mazhi,Longli
Y7 ÚmnmGoushan,Dushan Y27 cdBlJiangzhou,Duyun W47 t+(Tongzhou,Pingtang
Y8 cd~QFenghe,Duyun Y28 cdgh Mochong,Duyun W48 z{|} Mazhi,Longli
Y9 cdBlJiangzhou,Duyun Y29 cdgh Mochong,Duyun W49 ú@sYunwu,Guiding
Y10 cdgh Mochong,Duyun Y30 ú@sYunwu,Guiding W50 ú@sYunwu,Guiding
Y11 cdBlJiangzhou,Duyun Y31 cdBlJiangzhou,Duyun W51 tÎtDatang,Pingtang
Y12 ÚmnmGoushan,Dushan Y32 cdiBGujiang,Duyun W52 mFengshan,Fuquan
Y13 ú@sYunwu,Guiding W33 ´Ningwang,Huishui W53 t+(Tongzhou,Pingtang
Y14 cdgh Mochong,Duyun W34 Úm1mYingshan,Dushan W54 ú@sYunwu,Guiding
Y15 cdgh Mochong,Duyun W35 jc´kShuixiang,Shandu W55 Úm1mYingshan,Dushan
Y16 cdwxBaimang,Duyun W36 ´Ningwang,Huishui W56 Úm1mYingshan,Dushan
Y17 cdiBGujiang,Duyun W37 z{|} Mazhi,Longli W57 t+(Tongzhou,Pingtang
Y18 cdgh Mochong,Duyun W38
Najiang,Luodian W58
Najiang,Luodian
Y19 ú@sYunwu,Guiding W39 ´Ningwang,Huishui W59 ú@sYunwu,Guiding
Y20 cdiBGujiang,Duyun W40 ´Ningwang,Huishui W60 ú@sYunwu,Guiding
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10mmol·L-1dNTPs(0.5μL)、10×PCRÔJÕ
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(2.5μL)、2U犜犪狇DNA ¢×(0.5μL)、
ddH2O18.5(μL)。ØÆe:94 ℃ Ùn
5min;94℃n30s,51℃Ú60℃¸ÛÜ30s,
72℃ÝÞ30s,35ßØ;ßØ{फ72℃ÝÞ
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Table2 InformationofSSRmarkers
~$
Primer
"#f
Accession
No.
45Æ
Repeatmotif
~$ÆÇPrimersequence(5′→3′)
þ$Forward Ô$Reverse
ÛÜ%
Annealing
temperature
/℃
$Îø
Product
size/bp
QNSSR01 CV69972 (TTTTTA)3 GTCAAGAAAGCTCAAGGC AAGACCCATACAAAAGATACT 51 137~157
QNSSR02 CV067063 (TC)15 GGAGCATTGAAGCGAGAAAT ACGCTTCGAGTACTCCCTGA 55 165~199
QNSSR04 CV011305 (GGAAA)11 TAGCTCGCACACAACACCAC TCCAACGACACACTCTCTGC 58 170~204
QNSSR05 CV014525 (GTGGA)5 ATCCACCGTATGATGCTT TGTCTTGTGACCAAATTGAC 51 264~276
QNSSR06 FF682718 (CT)15 AAAGAGGAGAGGCGAGGACAG TTCAGGATACGCTTTGATGCC 58 85~136
QNSSR07 DN976198 (AAGAA)3 GCAGAAAACCCTGTCAAT ATCACCACCCCACCATAC 51 123~175
QNSSR08 FE942905 (CCGCCA)3 TGCCCACACCACGAATACGAC GAAGATGGTTGCGAATGGCT 57 115~135
QNSSR09 AJ621796 (CA)12 GCATCATTCCACCACTCACC GTCATCAAACCAGTGGCTCA 57 146~184
QNSSR11 AJ621798 (TG)13 CACATTGTGGCGTGTTATTAATTT ACATTGGCTATCTCTCATCATGG 56 285~287
QNSSR12 AJ621786 (GT)16 GAATCAGGACATTATAGGAATTAA GGCCGAATGTTGTCTTTTGT 53 171~237
QNSSR15 (ATG)10 GCGTCGTCCCTTCTTTCTAA GGGCAGCCATAACCACTACT 57 139~177
QNSSR16 (TTC)10 CAGGGTTGCAAGAAGTACCG ATCAACCGTATGGGCAAAAG 55 108~158
QNSSR18 (AG)14 GGGAGAACCAACCCAGTCTAT CCCAATCCGCTGTAGTAGGA 58 133~159
QNSSR21 CV013826 (CTC)5 ATGAGAAGGAGGACGATG CATTTATGGACCTGTTCG 51 122~124
QNSSR23 CV011305 (TG)8(AG)10 TGGATTCCACCCAGAGTCC CCACCGACTCGATGACATAA 57 130~174
91116 ABC,::;60vLMR0]^jklmniSSRoz
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Table3 ResultsandthepolymorphisminformationofSSRprimers
~$
Primer
No.of
genotypes
犖a
lPnÝ*
No.of
polymorphic
bands
lPnÝ*
o
犘犘犅/%
犖e
犎o
犎e
lPnV
)1
犘犐犆
Shannon
犐
QNSSR01 4 4 3 75.00 1.7337 0.5667 0.4268 0.3582 0.7174
QNSSR02 29 16 16 100.00 9.1837 0.4333 0.8986 0.8814 2.4256
QNSSR04 33 14 14 100.00 8.4606 0.8167 0.8892 0.8711 2.3343
QNSSR05 7 5 5 100.00 1.3213 0.0833 0.2452 0.2346 0.5587
QNSSR06 45 23 23 100.00 14.4869 0.6667 0.9388 0.9268 2.8622
QNSSR07 6 5 5 100.00 1.2746 0.1500 0.2172 0.2087 0.5049
QNSSR08 4 4 3 75.00 1.1456 0.1333 0.1282 0.1239 0.3099
QNSSR09 21 13 13 100.00 4.1190 0.4500 0.7636 0.7350 1.8695
QNSSR11 2 2 2 100.00 1.8967 0.0000 0.4768 0.3610 0.6657
QNSSR12 13 10 10 100.00 1.6412 0.3333 0.3940 0.3802 0.9723
QNSSR15 24 10 10 100.00 5.5771 0.7167 0.8276 0.7982 1.9148
QNSSR16 16 9 9 100.00 4.1119 0.7500 0.7632 0.7188 1.5922
QNSSR18 33 12 12 100.00 8.1630 0.8333 0.8849 0.8657 2.2564
QNSSR21 3 2 2 100.00 1.3846 0.2667 0.2801 0.2392 0.4506
QNSSR23 40 18 18 100.00 9.2784 0.8500 0.8997 0.8844 2.5319
Mean 18.7 9.8 9.7 - 4.9186 0.4700 0.6023 0.5725 1.4644
óTotal 280 147 145 98.64 73.7786 - - - -
0211 ! " # $ % & 36
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Table4 Thegeneticdifferentiationamongorigins
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Primer
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犉is
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犉it
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犉st
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犖m
QNSSR01 -0.5176 -0.3533 0.1083 2.0583
QNSSR02 0.2479 0.4866 0.3174 0.5378
QNSSR04 -0.1930 0.1451 0.2834 0.6321
QNSSR05 0.5722 0.6131 0.0956 2.3655
QNSSR06 -0.0743 0.1418 0.2011 0.9930
QNSSR07 0.0419 0.1883 0.1528 1.3857
QNSSR08 -0.1883 -0.0670 0.1021 2.1986
QNSSR09 0.1776 0.3798 0.2459 0.7668
QNSSR11 1.0000 1.0000 0.2912 0.6085
QNSSR12 -0.2341 0.1243 0.2904 0.6109
QNSSR15 -0.0576 0.1668 0.2122 0.9281
QNSSR16 -0.1761 0.0741 0.2127 0.9253
QNSSR18 -0.1773 0.0370 0.1820 1.1234
QNSSR21 -0.2840 0.0893 0.2907 0.6100
QNSSR23 -0.3045 -0.0564 0.1902 1.0644
Mean -0.0408 0.1936 0.2252 0.8601
Q5 TUW
Table5 MatrixofNeisgeneticidentity(abovediagonal)andgeneticdistance(belowdiagonal)among10origins
¶^
Origin
cd
Duyun
ú
Guiding
Úm
Dushan
´
Huishui
jc
Sandu
z{
Longli
Luodian
t
Pingtang
op
Wengan
Fuquan
cdDuyun 0.9439 0.8790 0.8348 0.6982 0.8700 0.8924 0.8474 0.7888 0.8491
úGuiding 0.0577 0.8748 0.8427 0.6931 0.9073 0.9062 0.8854 0.7754 0.8061
ÚmDushan 0.1290 0.1337 0.8213 0.7720 0.8439 0.8528 0.8413 0.8251 0.7759
´ Huishui 0.1806 0.1711 0.1969 0.7566 0.8324 0.8476 0.8613 0.7646 0.8412
jcSandu 0.3593 0.3666 0.2588 0.2789 0.6770 0.6430 0.6504 0.6916 0.6447
z{Longli 0.1392 0.0973 0.1697 0.1834 0.3900 0.8683 0.8289 0.8223 0.8137
Luodian 0.1139 0.0985 0.1592 0.1654 0.4416 0.1412 0.8872 0.8381 0.7506
tPingtang 0.1656 0.1217 0.1728 0.1493 0.4301 0.1876 0.1197 0.7230 0.7527
op Wengan 0.2372 0.2544 0.1922 0.2684 0.3687 0.1956 0.1766 0.3243 0.7807
Fuquan 0.1636 0.2155 0.2538 0.1729 0.4390 0.2062 0.2869 0.2841 0.2475
12116 ABC,::;60vLMR0]^jklmniSSRoz
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