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Phylogeny and Classification of the Moss Family Mielichhoferiaceae in China Inferred from Chloroplast DNA Sequences and Morphology

基于形态学特征和分子数据的中国缺齿藓科初步研究



全 文 :书!"#$%&
,2016,36(7):1351-1356
犃犮狋犪犅狅狋.犅狅狉犲犪犾.犗犮犮犻犱犲狀狋.犛犻狀.
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;<=
(1974- ),>,?@,ABC,DEFG#$HIJK。Email:lizyan74@sina.com
LMNO:;PQ,R,?@,BC,DE#$ST0$%JK。Email:limy@swu.edu.cn
,4犐犛犛犚56789:;<=>?@AB
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1,
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1,
! 1,!%(2
(1!(UVW% XU%Y,Z[650224;2!(W% X\XU%Y,2]400715)
C D:^ _ISSR`ab7cdefghi160cjkl,10bm$nklo101cpq,rst1upq97
c

vwxyzt1upq{|
(犘犘犅)}96.02%,~€(犐)‚0.4889,Neisƒ„t…u€(犎)‚0.3324。
hiwxz†‡ˆ

t1upq{|
(犘犘犅)‚33.51%,~€(犐)‚0.1943,Neisƒ„t…u€(犎)‚
0.1332。‰z†‡t…u(犎T)‚0.3239,cj†‡t…u(犎s)}Š0.1332。AMOVA‹ŒŽhi†‡S
W

hiƒ„‘犖m ‚0.3492。hizNeis†‡’“0.1481~0.4385。ƒ”UPGMA•–,—†‡’“
‚0.327˜,7chi•‚™š,›9šœUžŸ ¡、¢£、¤¥žŸ¦§¨hi©ª,›«šœ¬­Ÿ®£T
hi©ª

•–¯°±²hi†‡S³´µ’“¶·¸¹¯u

EFG

defg

†‡t…u

†‡S

†‡’“

º»¯°
HIJK$
:Q746+.5 !LMNO:A
犌犲狀犲狋犻犮犇犻狏犲狉狊犻狋狔狅犳犘犪狆犺犻狅狆犲犱犻犾狌犿犿犻犮狉犪狀狋犺狌犿犇犲狋犲犮狋犲犱犫狔犐犛犛犚犇犪狋犪
LIZongyan1,GUANMingyuan1,LIJing1,LIMingyang2
(1LandscapeandArchitectureFaculty,SouthwestForestryUniversity,Kunming650224,China;2HorticultureandLandscape
Colege,SouthwestUniversity,Chongqing400715,China)
犃犫狊狋狉犪犮狋:犘犪狆犺犻狅狆犲犱犻犾狌犿犿犻犮狉犪狀狋犺狌犿hasthehighornamentalvalueknownasslipperorchids,whichwas
restrictedlydistributedinkarstlimestoneareasofsoutheastYunnan,southwestGuizhou,northernand
westernGuanxiProvince.Thisstudyfocusedonthegeneticdiversityof7populationsfromSoutheast
YunnandetectedbyISSRmarkers.101bandsweregeneratedfrom160individualsby10primers,among
which97bandsbelongedtopolymorphicandeachprimeramplified10.1bands.Therewasahighergenetic
diversityindexatspecieslevel(犘犘犅=96.02%;犐=0.4889;犎=0.3324),butalowergeneticdiversity
indexatpopulationlevel(犘犘犅=33.51%;犐=0.1943;犎= 0.1332).Totalgenediversityreached
0.3239andgenediversitywithinpopulationsattainedto0.1332.Neisgeneticdifferetationamongpopu
lations(犌st)was0.1481-0.4385.Geneflow(犖m)was0.3492.Theseindicatedthattherewasahigher
populationdifferentiationamongpopulations.BasedontheUPGMAclusterdendrogram,thesevenpopu
lationswereclassfiedintotwogroupsatgeneticdistanceof0.327.FirstgroupincludedGulinqing,Magu,
Tianba,JiahanqingandYangliujing;SecondgroupwasmadeupbyDouzuiandXiaobazipopulations.The
clusterresultindicatedthattherewasnosignificantcorrelationbetweengeneticdistanceandgeographical
distance.
犓犲狔狑狅狉犱狊:犘犪狆犺犻狅狆犲犱犻犾狌犿 犿犻犮狉犪狀狋犺狌犿;geneticdiversity;geneticdifferentiation;geneticdistance;
bloodrelationship
  defg(犘犪狆犺犻狅狆犲犱犻犾狌犿犿犻犮狉犪狀狋犺狌犿)S¼
”(½(

¾¿!(Ÿ"À

Á!!ÀzÂÃqÄ
ÅÆÇÈ
[12],
„rÉÊË*zÌÍÎÏ

Ð20ÑÒ
80ÓÔÕÖ×Ø,vÙÚÛWÜÝÞß,ÝCITES
à‚9:HI#$

áâãä

N‚åæçè

éêëìíîïdefgðñ

‡ò0$%

óvŸ†‡t…uôJK
[39]。
def
gëìõö÷–Ÿøùú‡ò
[3],
Ðûü1ý

de
fg‹þ|ŸÿÕ|Ë!

"#Cò$vTÿÕ|
%W&}32.4%[4];„rÉÊË*zÌÍ.u,d
efgˆ³tX\()*vz+,

-¸Pz.
/01 犘.犿犪犵犻犮Lantern(defg × 23f
g
)[5];
defg#451t…

ç6789ËW

-
ç/

ç:

s;Ÿ<;zS°Ër=67
*
[6];
;>ô
[7]
S?4cdefghi†‡t…u,
‹Œ@[

r$vwxz†‡t…u*”hiwx

Ê20.3% z†‡t…uØДhizS,hi
†‡Swx*”7c(hizSRAPA
z‹Œ
[8];
B9CzDÐ¹¯JK@[

defg
ijs†‡89—DyzS¼EFGü

—H’
“I@Öo·¸zJ¹¯

—ËW’“I@Öo·
¸zK¹¯
[9]。
ISSR(intersimplesequencerepeat) L‚M
N2OPàȏklt1DNA ,þQRNMNS
T
[10],
ÐU5×Ø

—hi†‡t…uŸ†‡‹V
S?s·ŽïWoz,-
[1113]。
X½(zÄÅÆ
´ÈY&Z[fgz\Ûs]Ÿ^s]

_[`
IdefgÞabc%WzÈd

Þ_ISSRef
bg´Èhi†‡t…uBhS?

‚rHIij
µkõAÉÊÖþlm

1 noŸpq
1.1 PQ%R
rJKnoÞastuÇ

 ¯Ÿvwx3y
7chi,zhi{|}ޅ~,‚€¶u‚ƒ
cj

zcN4„…’“—2m ×y,7chin
160c…r(@2),…”H†‡sqˆþQ‰”
-70℃Š!†‹‡HŒ。
1.2 STUVW犘犆犚XY
Þ_Žz1.5×CTABqi„defgƒ„
©‰DNA,DNA0.8%‘’“”•–—˜$,
時30ng/μL—-20℃HŒš_。
þQs_zISSRm$[œ›œWž{ÂW
%
(UBC)íÕ6Ÿz77cm$,Dss„10bZ
 ¡kl

¢0pq£¤¥¦

t1u˧zm$©

,zPCRkl±¨
j°‚
:2.0mmol/L MgCl2、dNTPs0.40mmol/
L、©ªÙÜ«¬‚ 5.0ng/μL、1.25 U/20μL
犜犪狇DNA•<­、m$ÙÜ«¬0.35μmol/L。IS
SRPCRkl®P‚:94℃¯8u5min,94℃8u
45s,48~52℃°±45s,72℃²³90s,35c´
µ
,72℃²³10min,4℃HŒ。kl¢$2.0%
‘’“”•–—˜

1.3 Z[\]
pq¶Ÿ˜·Éʹ¸¹º|zkl»¼

{
½0/1¾¿,¨ _POPGENE1.32ÀÁ[14,15]Bh
t1Â3ÃS|
(犘犘犅)、̘ôƒ„(犖a)、Ê
Äôƒ„
(犖e)、Neisƒ„t…u€(犎)、
ShannonMŀ(犐)、ij‰z†‡t…u(犎T)、
ijIƒ„t…u
(犎s)、ijz†‡S°
(犌st)、Neis†‡’“(犇)Ÿƒ„‘(犖m)ŸÆ。Ç
ANeis†‡’“,^ _ NTSYSpc1.21[16]ÀÁb
hiBh UPGMAhiº»¯°S?。ST†
‡89zpÈS?
(AMOVA)_ GenALEXÀÁ
ŸÆ
[17]。
2 ‹Œ³S?
2.1 STUV^_
Þ_,zISSRPCRkl±¨j°,b77c
ISSRm$klÊ¢$zÊ42c,DsÉso10c
kl ¡uË*

t1u˧Ÿpq£¤zm$B
hhi†‡t…uS?

@1)。m$klpq%t
`1 abST5XYc_
Table1 Amplifiedresultsofthedifferentprimers
m$PÊ
Primercode

Sequence
(5′→3′)
klpq
No.of
bands
t1upq
No.of
polymorphic
bands
t1uÃS|
Percentageof
polymorphic
bands/%
UBC814 (CT)8A 9 9 100
UBC825 (AC)8T 10 9 90
UBC831 (AT)8YA 8 7 87.5
UBC834 (AG)8YT 10 10 100
UBC848 (CA)8RG 10 9 90.0
UBC855 (AC)8YT 12 12 100
UBC856 (AC)8YA 12 12 100
UBC862 (AGC)5 10 10 100
UBC840 (GA)8YT 12 12 100
UBC849 (GT)8YA 8 7 87.5
‰ŸTotal 101 97 96.04
2531 ! " # $ % &                   36Ë
`2 ,4犐犛犛犚Z[567897d:;5<=>?@eZ
Table2 Geneticdiversityindicesofsevengeographicalpopulationsof犘.犿犻犮狉犪狀狋犺狌犿
hi
Population
…r
Sample
̘ôÂ
ƒ„犖a
ÊÄôÂ
ƒ„犖e
Neisƒ„
t…u€ 犎
Shannons
€犐
t1u
ÃS|犘犘犅/%
 ¯ ¡ MGMG 22 1.3663 1.2539 0.1428 0.2094 36.63
 ¯¤¥ž MGJHQ 19 1.2871 1.1801 0.1020 0.1516 28.71
 ¯Už MGGLQ 25 1.4059 1.2682 0.1519 0.2241 40.59
uǬ­ WSDZ 22 1.3168 1.2383 0.1332 0.1928 31.68
uǦ§¨ WSYLJ 19 1.2871 1.2267 0.1220 0.1750 28.71
uÇ®£T WSXBZ 21 1.3465 1.2556 0.1430 0.2078 34.65
vwx¢£ MLPTB 32 1.3366 1.2474 0.1374 0.1996 33.66
hiPopulation 1.3352 1.2386 0.1332 0.1943 33.51
$vSpecies 160 1.9604 1.5529 0.3244 0.4889 96.04
Ì
:犖a.̘ôƒ„;犖e.ÊÄôƒ„;犎.Neisƒ„t¬;犐.Shannons€;犘犘犅.t1uÃS|
Note:犖a.Observednumberofaleles;犖e.Effectivenumberofaleles;犎.Neigenediversity;犐.Shannonsinformationindex;犘犘犅.
Thepercentageofpolymorphicbands
`3 67897d:;5犖犲犻狊<=fgh(bÍÎyp)W<=ij(bÍÎýp)
Table3 Neisgeneticidentity(abovediagonal)andgeneticdistance(belowdiagonal)of犘.犿犻犮狉犪狀狋犺狌犿
hi
Population
 ¯Už
MGGLQ
 ¯¤¥ž
MGJHQ
 ¯ ¡
MGMG
uǬ­
WSDZ
uǦ§¨
WSYLJ
uÇ®£T
WSXBZ
vwx¢£
MLPTB
 ¯Už MGGLQ  0.6778 0.8046 0.7460 0.7072 0.6768 0.7045
 ¯¤¥ž MGJHQ 0.3889  0.7758 0.7279 0.8623 0.7199 0.7845
 ¯ ¡ MGMG 0.2174 0.2539  0.7843 0.7452 0.6876 0.7343
uǬ­ WSDZ 0.2931 0.3176 0.2429  0.7400 0.7970 0.7847
uǦ§¨ WSYLJ 0.3464 0.1481 0.2940 0.3012  0.6450 0.7945
uÇ®£T WSXBZ 0.3903 0.3287 0.3746 0.2269 0.4385  0.7080
vwx¢£ MLPTB 0.3503 0.2427 0.3089 0.2424 0.2301 0.3453 
‚12p,‚m$855、856Ÿ840,tm$klpq
‚8~10p,Ê6cm$klt1ÃS|}100%。
2.2 67895<=>?@
10cm$nklo101cpq(@2),rst
1upq97c,t1pq{|}96.04%,zcm
$xÏkl¢010.1cpq。UžŸ ¡hi
t1uÃS|Ë*

SЂ40.59%Ÿ36.63%,¤
¥žŸ¦§¨hizt1uÃS|%®

ÑÊ
28.71%。7 chizÊÄôƒ„(犖e)—
1.1801~1.2682ҏ;Neisƒ„t¬(犎)8ӂ
0.1020~0.1519,Shannon€(犐)ϗ0.1516~
0.2241。®£TŸ¢£hiz†‡t…u¹ÈÔ
W

¤¥žhiIz†‡t…uË!
(犘犘犅=
28.71%;犎=0.1020;犐=0.1516)。Užhi
z†‡t…uwx%*
,Shannon € (犐)‚
0.2241。—hiwxzShannon€Ÿ Neisƒ
„t¬‚0.1943Ÿ0.1332,—$vwxzSЂ
0.4889Ÿ0.3244。‰z†‡t…u(犎T)‚
0.3239,hiIz†‡t…u(犎s)‚0.1332。
2.3 :;k<=ijlmnEo
õA·ŽzhizNeis†‡’“(@3),Õ
’“hiz®£TŸ¦§¨z†‡’“%W

‚
0.4385,Ö¤¥žŸ¦§¨hiÊ%®†‡’
“

‚ 0.1481,7 chi  x Ï † ‡ ’ “ ‚
0.2992。ƒ”UPGMA•–(×1),—†‡’“}
0.327˜,7chi•‚™š,›9šœ™cSš©
ª

›9SšœUžŸ ¡hi©ª

›«Sšœ
¢£

¤¥žŸ¦§¨hi©ª

›«šœ¬­Ÿ®
£Thi©ª

‹ŒØþïhi†‡’“³´µ’
“¶·¸¹¯u

2.4 6789:;k5<=Jp
ij†‡S°[[Ù܆‡t…uz8
9ØÛz€`
[18],
†‡Sz*!@[hiz8
9ŸSwx
[1920]。犌st、犘犺犻犘犜 [Ô¸ÀÁŸÆ
35317Ú              ;<=,ô:ƒ”ISSRzdefghi†‡t…uJK
×1 defg7chizº»¯°•–
Fig.1 Dendrogramanalysisshowingrelationships
amongsevenpopulations
z†‡S€

ÛÜ
,7chiz†‡S°(犌st)}
0.5888,ŽïhiŒ—Ë*z†‡Swx。
AMOVAŽÊ39.60%zST†‡89Ø۔h


Ý9‹ŒÞ*”ßàUô&áz‹Œ
[21],
®
”³犌stzSwx,³犘犺犻犘犜ÏÕ¹¸。Öhi
xσ„‘
(犖m)т0.3492。


¸ 9 ´ È  h i z † ‡ S  ° 
(犘犺犻犘犜)¹ÈËW,uÇ®£T³¬­z犘犺犻犘犜
ς0.304,¬­³¦§¨z‚0.430,®£T³¦
§¨z†‡S°}0.492,xφ‡S°
}0.409;Ö ¯ ¡ŸUžÑ‚0.319,U
ž³¤¥žz犘犺犻犘犜 ‚0.472, ¡³¤¥žÒ
‚0.381。¢£hi³rã6chi犘犺犻犘犜 Ï
—0.338~0.413ҏ,³¬­S%®,³Už
%W
。7chi™™†‡S%®z‚¦§¨Ÿ
¤¥ž

}0.278;%Wz‚¦§¨Ÿ®£T,}
0.492。7chixÏ犘犺犻犘犜 ς0.392。hi
犘犺犻犘犜 ϱ²oz†‡S®¬³ Neis’“
¯°[9äz

å榧¨Ÿ¬­hi‚Õ»h
i

çhi™™zxφ‡S°èË*

é[
¹¸´ÈՒ“hi_Œ—Ë*z†‡S

±
²ïhi†‡Swx³´µ’“¶[·¹
¯u

3 ê k
3.1 6789:;5<=>?@
9ëì‚

Á¼ví{î¼vÉÊË*z†‡
t…u

FG#$ÊË!z†‡t…u

†‡t…u
ï!z#$ïFG
[2223]。
çïØïtzJK@[

FG#$ðÊñ*z†‡t…u

Þ_ISSRef
bòóô
(犌犾犲犺狀犻犪犾犻狋狋狅狉犪犾犻狊)、õ:öç÷(犆犪犿犲犾
犾犻犪狆狌犫犻狆犲狋犪犾犪)FGv†‡t…uwxz—˜·
Ž

—vŸhiwxíHøË*z†‡t…u

hi
†‡SðÔ*
[2425]。
Þ_ISSRef—˜g4FGvzJK@[,
‘ùÄú
(犇犲狀犱狉狅犫犻狌犿犳犻犿犫狉犻犪狋狌犿)—vwxz†
‡t…uÔ!

çhiwxz†‡t…uË!
[26]。
r78b7cdefghizISSR—˜‹Œ@Ö
o$vwxz†‡t…uË*
(犘犘犅=96.04%,犎
=0.3244,犐=0.4889),hiwxz†‡t…uÏ
*”yûv
(犘犘犅=33.51%,犎=0.1332,犐=
0.1943)。$v‰z†‡t…u(犎T)‚0.3239,
hiIz†‡t…uт0.1332。ü‹k³
SRAPAŸ;>zýC—˜‹k[9äz[89],þ
defgvwx†‡t…uÿ*”hiwx

ç³
ßàUô&ázË*wxzhi†‡t…u
(犘犘犅
=80.28%,犎=0.2847,犐=0.4236)‹ŒÊËWo
!
[21]。
åæFG$z†‡t…u³ñt„"ʯ

-
D#ÜÁ¼vz$vsSÖØ

S¼$%

0&p
Á

ðó°¶ô
。Hamrick[22]ì‚hi†‡89W
®zëì(„"õâ‚ðó°¶

S¼$%Ÿ0
)*

Á¼

9,Ÿö+$‡,vTz$v--ÉÊ
Ë*z†‡t…u

defgZHøvË*wxz
†‡t…u³r9,

./0r

vT‡,.u

S
¼0&zt…12¹¯

3.2 6789:;5<=Jp
9ëì‚
,犘犺犻犘犜S€3—0~0.05ҏ
é[hiSñ4

3—0.05~0.15ҏé[h
iŒ—sô®¬S

3—0.15~0.25ҏé[
hiSW

3犘犺犻犘犜 W”0.25@[hiS
Y W
[27]。7 c d e f g h i z S  € 
(犘犺犻犘犜)xÏwx}0.392,5Sé[defgh
iSW

³ AMOVA ‹ŒÕ¹¸,!”ƒ”
Neis€z犌st(0.5888),6Ք¹¸´È78Ä
úhi犌st(0.5637),ÿ*”defg—¾9
,:Èz犌stwx[21]。ƒ”defgzISSRST
`a
,^
_Ô¸zS?pq
(犘犺犻犘犜,AMOVA);

JK@[

9,–iz
x󄆇S°犌st‚0.23,=,–i犌st‚
0.19,Ð,–i犌st‚0.59[28]。defg>”9ç
‡ò#$

hiz†‡S·¸

$vzðó°¶[(hiwxyz†‡S
wxz2ì„"
,Avise[29]삜4?hZcz+
$CòŸövT¹ºz#$hiҏz†‡S·
4531 ! " # $ % &                   36Ë
¸。
defgëìœ÷–‡ò
[3],
r?h’“Ê@

ëì[BhhiIíç#4ҏzCò

Aª†‡
89ëìØ۔hiI

Öhizçò‘[BC
Ê@z

DO

œ”defgvTÿÕ.uÉÊEF
z0&st.3

äGä$v´µHuS¼


hiz´µ’“Ëÿ

9ëì‚
,´
µI“Jm
\Õ,

ÖÕ,ðñpK·m\Õ,L°ŸM!i
jz†‡t…u
,¸
…ðJM!ijƒ„,‘

›NijzS
[3032]。
hi†‡%JKs

ƒ„
‘ÇA 犖m zW®OS‚*(≥1.0)、s(0.250~
0.99)、!(0.0~0.249)Pcô:wx[33]。3{ü
`QOS

üâJKzdefghiƒ„‘R”
!wx

т0.3492,¹¸zISSRefŽz
(

¾¿ŸÁ!,:Èdzdefgzƒ„‘ðÑ
Ê0.7201[21]。9ëÖS,—stN_Ô[·˜,T
ìÊñUÜzƒ„‘V&ÊÄ´WâijS
[34],
&XHƒ„‘Ô[Aªdefghi†‡S·
¸zëì„"

ÊïØïtzJKØ[

†‡89ëì[œs
t

W8

ƒ„‘Ÿ†‡Y8ô¢0

ÖD’“Z
Ô9¡¢0†‡89
[35]。
[ü¹’ñÕzijþ
\Œ—]ðzƒ„‘+

EÊczstN_ðZ
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