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,2016,36(4):0674-0680
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犪狀犱犌犲狀犲狋犻犮犛狋狉狌犮狋狌狉犲犃狀犪犾狔狊犻狊狅狀犖犪狋狌狉犪犾犘狅狆狌犾犪狋犻狅狀狊狅犳
犈狀犱犪狀犵犲狉犲犱犚犺狅犱狅犱犲狀犱狉狅狀犿狅犾犾犲犌.犇狅狀
CHENGMalong,DAILiangfang,LUOXiangdong,LIUMenglu,ZHOUYi,
CHENGYanqiu,XIEJiankun
(ColegeofLifeScience,JiangxiNormalUniversity,Nanchang330022,China)
犃犫狊狋狉犪犮狋:Inthepresentstudy,12pairsofsimplesequencerepeat(SSR)primerswereusedtoevaluatege
neticdiversityandgeneticstructureof犚犺狅犱狅犱犲狀犱狉狅狀犿狅犾犾犲G.Donineightpopulationsfromsixprov
inces.Theresultsshowedthat:(1)Atotalof260aleleswithanaverage21.667alelesperlocusweream
plifiedfrom193individuals.Averagevaluesofnumberofeffectivealeles(犖犲)andpolymorphisminforma
tioncontent(犘犐犆)were5.425and0.900,respectively.Thisspecieshashighergeneticdiversityandgene
flowamongdifferentpopulations.For犚.犿狅犾犾犲,theShannon’sinformationindex(犐)andNei’sgenediver
sity(犎)were1.768and0.777,respectively.(2)ComprehensiveanalysissuggestedthatJinxi(Jiangxi
Province)populationexhibitsgreatlevelofvariability,whereasthepopulationofZhenghe(FujianProv
ince)exhibitsthelowestlevelofvariability.(3)Thegeneticdifferentiationcoefficient(犉狊狋)basedoninfi
nitealelemodel(IAM)was0.142andthegeneflow(犖犿)was1.522,respectively.Analysisofmolecular
variance(AMOVA)revealedthatthegeneticvariationmainlyoccurredwithinpopulations(accountedfor
87.71%),morethaninterpopulations(accountedfor12.29%).(4)Theresultsofneighborjoining(NJ)a
nalysisbasedongeneticdistanceandstructureanalysisshowedthateightpopulationswereclusteredinto
threegroups.Themanteltestshowedtherewasnosignificantcorrelationbetweengeneticdistanceandge
ographicaldistancein犚.犿狅犾犾犲.Ourresultsindicatedthepriorityshouldbegivento犻狀狊犻狋狌conservation,
naturalpopulationsinJinxi,Zhenghe,andJingshan(HubeiProvince)deservepriorconservation.Thepossi
bleformationofthehighlevelgeneticdiversityandthemoderatediferentiationof犚.犿狅犾犾犲werealsodiscussed.
犓犲狔狑狅狉犱狊:犚犺狅犱狅犱犲狀犱狉狅狀犿狅犾犾犲G.Don;SSR;geneticdiversity;conservationofendangeredplant
abc(犚.犿狅犾犾犲),ÑÒÓÔÕ、Öa×,Ø
ÎÔÕ×+abcÙÚÎÛg1`y:[1],
ÜÌÍgÝÞßà
,
ØÔÕ×áâãägÉ<
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A
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、
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、
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xcdef`$g¾GCgxhkX
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(Simplesequencerepeat,
SSR),xyz4 {|,z}Î,Üj、Ív
Éuj
、
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Ø89efMNhij
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[910]。
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NhijkMNlm
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VW}-
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MNhijkMNlm
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±²³´
[11]
«~ñabcµ¶,z}
DNA。BW1%·¸¹º»¼½k¾èA¿¿v
²³DNAiâgÀvkÁv,-20℃Â|zÃW。
15μLgPCRÃf¥ÄÅ7.5μL2×犜犪狇PCR
greenmix("ÆÆUÇ:$ÈÉ¢ÊË~),
ÂÌSSR
$(®2)«1.0μL,ddH2O4μL,,
z} DNA1.5μL。d¾ÍÎ{:94 ℃Ïj
3min;94℃j45s,55℃(ÐA
$56℃)ÑÒ
45s,72℃uj1min,35`ÓG;72℃ÔÕ7min,
¦4℃|z。ÃÖ$¼20bpDNAladderE
{AB\]
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48%%j¬×ØÙÚº»Î¼
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ÛÜ¢{1×TBE(pH8.3)。150VÝü¼½
90min,Þßßá[12],4à©á²。
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âABQøãfX,z
,
]ªÍ
gä,Qø
(
åæ
,
ABQø{120bpgÁ
Íw]{120/120;AB{120bpk126bpg
çÍ]{120/126),Íèé]{“0/0”,êJ
SSR®¤。ëÀ,VW GenALEx6.1ìÎ[13]
í«MNhij
;
VW POPGENE í
Nei’shij(犎)、MNAu¥(犉狊狋)_,z¦
(犖犿)[14];ðî,VWGenALEx6.1ìÎÎgAB
a1 678;
de>æ
Populationslocation
i®
Samplesize
¬v
Longitudeandlatitude
ïð
Altitude/m
34-Jinzhai,Anhui(JZ) 21 115°47′E,31°11′N 506~706
2.3Pan’an,Zhejiang(PA) 24 120°27′E,28°52′N 514~906
.! Yongxiu,Jiangxi(YX) 21 115°43′E,29°08′N 16~39
Å"ÆÇJingshan,Hubei(JS) 22 112°50′E,30°59′N 80~164
.!-Jinxi,Jiangxi(JX) 30 116°45′E,27°50′N 84~100
ÅTIShaodong,Hu’nan(SD) 33 111°50′E,27°04′N 209~321
.!Poyang,Jiangxi(PY) 24 116°57′E,29°16′N 67~104
kZhenghe,Fujian(ZH) 18 119°05′E,27°16′N 993~1176
5764
012,:ÏÐabc)*degMNhijkMNlmAp
ñ
(AMOVA)òxnfóApMN4abc
de¨
、
de§gMNAust
;
ô¦BWPower
MarkerV3.25ìÎX«Aph?jD
(犘犐犆),íMN©ª(犇)kMNfõv(犌犐);
öW Neighborjoining(NJ)í«m¬÷,øV
WGenALEx6.1í¶·©ª,¬Mantel²³,A
pð@MN©ªg¹ºj
。
¦BW STRUC
TURE2.3.4ìÎApeflm[15],ãùef
} 犓,犓 ñ1~13,10îÉu,BW Structure
Harvesterí△犓 àE{ù犓 àú\û[16]。
2 l@Ap
2.1 678犛犛犚cd@eB
®89VW12`h?j
$Xabc8`)
*de193``fnoSSRAp,ü260
`,z
,
t`,z{21.667,
Ãýþÿv490~182bp!¨。®89"Wg
12XSSR
$h?jÌ»,ÃÍ#$,æ÷1
Î
$ HD88XIefÃl"。Ap®
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%ÁÃg,zñÌQ
,
ðÎ
8gØ HD83,{14`,hgØ HD34、
HD39k HD64,{26`(®2)。12`gh
?jD
(犘犐犆)40.790~0.941,tà{
0.900,abcMN&guçj。«gM
NAu¥ 犉狊狋àñ$̯,( 0.067~
0.251,tà{0.142,®¯de§Q de
¨
。
2.2 6789:;<=>?@AB
MNhijApl
(
®3),8`abc)
*deÎg,z{4.087~6.905,t
{5.425,ðÎÍ{-(JX)de,k(ZH)d
e
。Nei’s,zhij(犎)uP){
0.706~0.820,tà{0.777,8`degMN
(Nei’s,zhij)*v;JÍî{k
(ZH)<ÆÇ(JS)<(YX)<(PY)<-
(JZ)<3(PA)<I(SD)<-(JX)。Shan
non(犐)tà{1.768(1.493~2.009),ðM
Nhij*v+v@Nei’s,z(犎)gl,
M.20bpDNAladder
÷1
$ HD88XIdeÃl
Fig.1 AmplificationproductsofShaodongpopulationbyHD88
a2 67812f犛犛犚cd>?@AB
Table2 Geneticdiversityof12SSRlociin犚.犿狅犾犾犲
SSR
SSRlocus
,z
Numberofaleles
(犖犪)
,zhij
Genediversity
index(犎)
MNAu¥
Coefficientofgene
differentiation
(犉狊狋)
,z¦
Geneflow
(犖犿)
h?D
Polymorphism
information
content(犘犐犆)
HD25 21 0.924 0.067 3.479 0.922
HD26 17 0.883 0.223 0.873 0.877
HD34 26 0.935 0.112 1.975 0.939
HD35 24 0.929 0.095 2.379 0.925
HD37 15 0.812 0.162 1.295 0.790
HD39 26 0.942 0.146 1.450 0.941
HD43 25 0.926 0.172 1.201 0.925
HD55 23 0.919 0.163 1.287 0.917
HD64 26 0.932 0.076 3.040 0.932
HD80 21 0.889 0.251 0.745 0.881
HD83 14 0.864 0.150 1.420 0.853
HD88 22 0.906 0.090 2.537 0.901
t Mean 21.667 0.905 0.142 1.522 0.900
676 ! " # $ % & 36,
®fõ
(
®3)。-«MNhij±gApl
®¯
,
-
(JX)¶7abcef§.ÌgM
N
,
I
(SD)、3(PA)î!,-(JZ)、
(PY)、(YX)、ÆÇ(JS)4`¶7deMNhi
jstdÎ
,
k
(ZH)deMNhijst。
2.3 6789:;<=>?DEgh
2.3.1 6789:;<=犃犕犗犞犃FG abc
8`)*deMNAustgABñApl®
¯
(
®4),abcefgQÐAz4 de§
(87.71%),de¨gMN{12.29%(犘<
0.001)。/l@;Nei’sígef¨Au
¥
(犉狊狋=0.142)²³glfõ。ÁÂ,abc
det,z¦犖犿 {1.522(®2),®¯abcd
e¨wxC0ç12,z34Ö:fo0vg,z
1¦
,
=567ËzMN8
9gde¨A
u0v
。
2.3.2 678;?jk`犖犑lOFG
MN©ª
(犇)kMNfõv(犌犐)Ap®¯(®5),
abc«deNei’sMN©ª_MNfõvgt
àA:{0.1426k0.8662,®¯«de;<º¥
5Ì=
。
2.3
(PA)@ÅTI(SD)deMN
fõvQ
,
{0.9054,ÁÂMN©ªø,{
0.0994。.!(YX)@k(ZH)def
õvø
,
{0.8055,>F¨MN©ªQ,{
0.2163。¬ Mantel²³´µ,abc«de¨g
MN©ª@¶·©ª¡¸¹º
(狉=-0.164,犘=
0.248)。
%Áabcde"mg NJ¬l(÷2)
®¯
,8`de;(JS)de{Ⅰ,-(JZ)、-(JX)、k(ZH)
k
(PY)de¬{Ⅱ,3(PA)@I(SD)2
`de¨MN©ªø
,
º¥?@
,
¬{Ⅲ。
2.3.3 6789:;<=犛狋狉狌犮狋狌狉犲FG , A
¶Bí«gStructurel,abc«deµ
ËfogMNAu
(
÷3)。△犓 Qà"X¾g犓
à{3,®¯abc)*deStructureApgg
犓 à{3。StructureApl,abc8`d
ewAî3Q},A:W3Cá>®(÷4),w
a3 6789:;<=>?@AB
Table3 Geneticdiversityofnaturalpopulationsof犚.犿狅犾犾犲
de
Population
,z
Numberof
aleles(犖犪)
,z
Effectivenumber
ofaleles(犖犲)
Shannon
Shannon
index(犐)
,zhij
Genediversity
index(犎)
Do
Fixation
index(犉)
JZ 7.58 5.061 1.724 0.783 0.415
PA 9.00 6.010 1.893 0.805 0.420
YX 8.83 5.581 1.791 0.770 0.402
JS 7.25 4.720 1.634 0.754 0.330
JX 10.58 6.905 2.009 0.820 0.367
SD 9.25 6.029 1.871 0.806 0.551
PY 7.83 5.004 1.728 0.771 0.449
ZH 6.48 4.087 1.493 0.706 0.188
t Mean 8.35 5.425 1.768 0.777 0.390
E:de,FÁ®1;ÂÁ。
Note:Populationcodesaresameastable1;Thesameasbelow.
a4 678>?Fmno=pqFG(AMOVA)
Table4 AMOVAofthegeneticdifferentiationlevelin犚.犿狅犾犾犲
>æ
Sourceof
variance
);v
犱犳
ñk
Sumof
squares
tñ
Meanof
squares
}A
Variance
component
GAH
Percentageof
variation/%
犘
de¨Amongpopulations 7 278.334 39.762 0.683 12.29 <0.001
de§ Withinpopulations 185 1823.511 9.738 4.869 87.71 <0.001
üTotal 192 2101.845 5.552 100.00
}¨Amonggroups 2 175.104 87.552 1.153 6.86 <0.001
}§ Withingroups 190 2973.585 15.650 15.650 93.14 <0.001
üTotal 192 3148.689 16.803 100.00
E:犘à®5ݳàQgIH,J`IHØC0KdeÎgi®¬01000îôL*wãíêJg。
Note:犘valuesaretheprobabilitiesofhavingamoreextremevariancecomponentthantheobservedvaluesalone.Probabilitieswerecal
culatedby1000randompermutationsofindividualsacrosspopulations.
7764
012,:ÏÐabc)*degMNhijkMNlmAp
a5 犖犲犻’狊>?jk(±M)` >?rst(±M)
Table5 Nei’sgeneticdistance(belowdiagonal)andgeneticidentity(abovediagonal)
de
Population
JZ PA YX JS JX SD PY ZH
JZ - 0.8847 0.8341 0.8572 0.9041 0.8886 0.8703 0.8671
PA 0.1225 - 0.8608 0.8713 0.8969 0.9054 0.8685 0.8521
YX 0.1814 0.1499 - 0.8442 0.8747 0.8559 0.8606 0.8055
JS 0.1541 0.1378 0.1694 - 0.8700 0.8718 0.8593 0.8173
JX 0.1008 0.1088 0.1339 0.1393 - 0.8937 0.8529 0.8529
SD 0.1181 0.0994 0.1556 0.1372 0.1124 - 0.8838 0.8647
PY 0.1389 0.1410 0.1501 0.1516 0.1210 0.1235 - 0.8858
ZH 0.1426 0.1600 0.2163 0.2018 0.1591 0.1454 0.1213 -
÷2 abcef¨NJ¬Ap
Fig.2 NJanalysison8populationsof犚.犿狅犾犾犲byPowerMarker
÷3 VW△犓XeflmnoAp
Fig.3 Thegroupstructureanalysisonbasing△犓
1~8A:{JZ、PA、YX、JS、JX、SD、PYkZHde;¹ÁgCá,®f`e。
÷4 WStructureNOg8`abcdeMNlm(犓=3)
Thenumber1to8representedJinzhai,Panan,Yongxiu,Jingshan,Jinxi,Shaodong,Poyangand
Zhenghepopulation.Thesamecolorindicatesthesamepopulation
Fig.4 Estimatedgeneticstructurefor犓=3obtainedwiththeSTRUCTUREprogramfor8populationsof犚.犿狅犾犾犲
¼P
,
-
(JZ)、-(JX)、k(ZH)de¬{
f}
,
3
(PA)、I(SD)、(PY)de¬{f
}
,
(YX)、ÆÇ(JS){f}。nfó5ÌAp
´µ
,StructureApl@, MN©ª«g NJ
¬l,®fõ
,
QR
(PY)deè,ðS
7`deg¬lTéfõ。nfóX8deA
{3}noAMOVAAp´µ,abc}¨MN
{6.86%(®4),}¨tAu¥Φ狊狋{0.069
(犘<0.001),UîV¯abc)*de}§Q
}¨
。
3 r Ê
MNhijØ$ÿ
:zk|jnulhg
$W,X
,
Ø:$hij|}gÉ<}îÐA
。
f
Y5Z
,
、
ÏÐgMNhijstÌ
[17],
:$Wx|jÌÍstgMNhij
[18]。
ab
cØÎabcÙÚÎÛgf`y:
,
; ð
ïWßàk[\ÝÞßà
,
<{?@]î:CDE
,
876 ! " # $ % & 36,
A9^_Tµý`u
,
aOõabcî{ÏÐ
[19]。
®89Î
,12XSSR\]Apabc
ÜÌÍgMNhij
(犘犐犆=0.900,犐=1.768,
犎=0.777),Í ç-VWISSR\]Ap.!a
bcgMNhij
[20](犐=0.5007,犎=0.3342)¼
_ÁÚgbcÔÕ
[21](犐=0.4972,犎=0.3386),d
SSR\]Apg66`ÔÕâMNhij[22]
(犘犐犆=0.838,犐=2.2293),9ðyz,fewx
89g)*deQø_:?¤%Á"õ
;
ff
ewxØabc{h+:
、
gRÌÿ#$
,
y®A9
P)Ì{xy
,
ÿ
gdehukÔkSiðjk
ÌhMN
,
lئ
0vB@_ð:CmDE
µý`u
,
:ý`u¨%ÿ
,
MNhij(4n
éoþ
。
ý`u:CXegMNlmkMNhijs
tÖ:pQLM
。
fY5Z
,
:Cý`uq
deQø
,
]î$g¾GCxhÂq
,
ÁÂ$wx
rz{MN8k=1sÑ5ide¨Ö:MNA
u
,
n5q/eg:zxh
,
;ç4¦,Ö:M
Ntu
[23]。
v=g89
,
abce¨Au¥
犉狊狋{0.142,ÚÎ0vAu[24],wMN;
fÎ87.71%´: de§,de¨MN
Û{12.29%,, StructureApA{±}¦g
AMOVAAp$de§MNQ de¨
MN
。
x\®¯
,
abc¬ydeý`u
!¦de§TÎ0vgAu
,
9ðyzwxØd
e¨zz4fo0v,z¦
。Slatkin[25]{{|犖犿
>1Â,,z¦w¼}~de¨zMN
9g
deAu
,
®89Îabcde¨犖犿 {1.522,®
¯LMabc8`deMNlmg;
,
5×kBgnØ#$,z¦;<>
[26]。
âÔÕ×+×ÜjkBø
Ü
[22],
N³"kýØÌ©ªN
,z¦g;<
。
çè
,
A97zB
、
:Chij_<
z
,
XMNhij_MNlm£Ö:
ÌQLM
[27]。
v=N趴µ
,
abc;<
A94ïð20~1200mgÇ2Ç,«/
abcde"(:CpQ%Á
。
I
(SD)、k
(ZH)_.!3`deÚ %|}7§de,h(
Sg
,
h@+
、
ÔÕ+:
,
<
、
@z ¡5̯
,
õiabc
de¢Q
,
£Âgh¼¤nA9{;
。
ÆÇ
(JS)、-(JZ)、3(PA)3`de(4¥¦)*
|}7
,
h@§+
、¨
+©:
,
)*de:C
DEÌ8
,
:$mJË%Á0v<{zgDE
。
zçXabcN:deg|}_ªÉ«
。
®89g
StructureApk NJ¬Ap®¯,8`dew
¼A{±Q
,
(YX)、ÆÇ(JS)¬{f
}
,
3
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