应用SRAP标记对西藏特有植物大花黄牡丹的遗传多样性进行研究。用16对引物从5个自然居群79个单株中共检测到396个有效位点,其中多态性位点357个。在物种水平上,多态位点百分率(Ppl)为90.15%,Shannon表型多样性指数(Ηsp)平均为0.252 1; 居群水平上的Ppl为31.82%,Shannon表型多样性指数(Ho)为0.069 4~0.342 8,平均值(Ηpop)为0.130 7。上述遗传参数表明,大花黄牡丹具有丰富的物种遗传多样性,5个居群中自然居群C的遗传多样性最高(Ppl=82.32%,Ho=0.342 8)。据AMOVA分析结果,总的变异中有41.58%的变异存在于居群间,58.42%的变异存在于居群内,居群分化较显著(ΦST =0.415 8,P<0.001),由POPGENE1.32得到的居群间遗传分化系数GST(0.430 9)和Shannon表型多样性指数计算的居群间遗传多样性所占比例(0.481 6)也表明了类似的遗传结构。Mantel检测表明地理距离和Nei‘s遗传距离间相关不显著(P>0.05)。利用NTSYSPC(2.1)软件构建大花黄牡丹5个居群79个个体的UPGMA聚类图,遗传相似系数变幅在0.47~0.99,大多数居群内的个体表现出较为密切的亲缘关系(如居群B,D,E),但也有一些居群的个体未聚在一起(如居群C)。依据大花黄牡丹居群遗传变异特点,初步探讨其保护和利用策略。
Sequence Related Amplified Polymorphism (SRAP) analysis was applied to study the genetic diversity of Paeonia ludlowii, a species only distributed in Tibet of China. Sixteen pairs of SRAP primers were used to evaluate the genetic diversity of 79 samples from five natural populations. A total of 396 valid loci were obtained, of which 357 were exhibited polymorphism. The percentage of polymorphic loci (Ppl) and the mean diversity estimated with Shannon‘s index of phenotypic diversity(Ηsp) at species level were 90.15% and 0.252 1, respectively.At population level, Ppl was 31.82% and Shannon‘s index (Ho) ranged from 0.069 4 to 0.342 8, with an average value(Ηpop) of 0.130 7. Among the five populations, population C was especially more diverse(Ppl=82.32%,Ho=0.342 8). Those results indicated that there was a relatively high level of intraspecific genetic diversity. Analysis of molecular variance(AMOVA) demonstrated that a high degree of genetic differentiation occurred among the populations (ΦST =0.415 8,P<0.001). The proportion of genetic variation among populations was 41.58%, while the proportion within population was 58.42%. A similar analysis result was exhibited by the genetic differential index(GST) of 0.430 9 and the proportion of diversity among populations with Shannon‘s index of 0.481 6. Mantel test showed that there was no significant correlation between geographical distance and genetic distance (r=0.597 5,P=0.905 5) among populations. A dendrogram conducted by UPGMA method using NTSYSPC(2.1) demonstrated that the range of genetic similarity coefficient was 0.47-0.99. The most of the individuals came from the same population had close genetic relationship, such as population of B, D, E,but individuals from population C could not be clustered together completely. A strategy for the conservation and utilization of P. ludlowii based on genetic diversity was proposed.
全 文 :!!"
" !
#
#
$ % # $
$
#
%
& ( )
&()*+(, &(-.,) &(*(,)
./01!"!*/1#
2345!$ % # $
E`æW*ÈöÛK;>
&E,?
[!
7
#
7
`
$!8
7
=
#
7
¡
8
7
¢
7
£
$
7
¤¥¦
$
7
§¨©ª
$
"#1
fgCE)¯°)F
7
e
>$:%#!# $1
AqÒ×C¢()FXYPQ§
7
Ô
":%%8$#
81
fgCE)x}PQ§
7
e
>$:%#!$
!
7
"
!7
Þ¹
&E,?
å=Aqç{§¨E`æW*>ÈöÛK;öPQ
%
¹
#>
=|¨Ã
:
¢%
9;
¢3gRÛNÁ
8;>
¢{í
!
ªRÛá;í
8:9
¢
%
|¨¬Qò
!
Ûáí¼[
"!
@0
$
f
;%1#:i!
&L344/4
_XÛK;ÜÚ
"
V@
$
òif
%1$:$ ##
%Qò>
!
@0
f
8#1"$i!&L344/4
_XÛK;ÜÚ
"K
/
$
f
%1%>; ! %`18!$ "!
òi
"
@/@
$
f
%1#8% 9%
ºÈöÚ_`
!
E`æW*·{uv>¨¬ÈöÛK;
!:
¢%
R%
>ÈöÛK;St
"!
@0
="$18$i!K
/
=%18!$ "$%
1
,\h.,
[\]^
!
ÿ>YmR{
!#1:"i
>
YmË|u%
!:"1!$i
>YmË|u%s
!
%[:dop
"
(
&+
=%1!#: "!!f%1%%#$!
°
?h?K)*)#18$
WÁ>%Èö[:Ú
_
&+
"%1!8% ;$
&L344/4
_XÛK;ÜÚ¡£>%ÈöÛK;§ìÖ»
"%1!"# >$
1_`ßW¸>Èö]
% \34TF0
ÛN_`3,J²
*FIoV
ÈöJ²vwnop
"!m%1%:$%
x¹
*+&C&
?
"$1#$
PÅE`æW*
:
¢%
9;
¢¢ú>
k?K\,
òWß
!
Èöv¸ÚYT|
%1!9 %`1;;!
EÛ
Ú%s>¢ú_rÝdfb>äîw
"
o%
D!P!)$!
h1{Ê$%>¢ú$ò|Ê}
"
o%
$%
0
1E`æW*%ÈöYmç
!
p_*)ª;<x¹Ú¯
%
#$%
&7
E`æW*
# &E,?#
%
#
ÈöÛK;
#
Èö[:
&()*
! &9#"1!>% &9#"1:!777
+,-./
!,777
+01*
!#%%# 69!"""$%#$#%# 6%%9% 6%9
#
& $%## 6%8 6###
#
& $%## 6## 6$$%
&
AqÒ×ZÛ
"
qZÃ<
$%%;"%#$
Í
$#
fgCE)®¯É
%
!`f¢£¤¥
%
5$;9;&6>0)0*2W#()3S)I#"0)(> *20$27# -,=-2?#)&!)<#(
+34JMI4
#
7RF4JGIH0I
$!8
7-I3/\I4Jo34
#
7?34 KH34JT34J
8
7RL3GI
$
7K/4J2H4LH3
$
7ISF4 RLH/J3
$
"#1#$%&&$(K$".5:;%.;"&! ?5:/;8* )"5:;%.;"8%4*56&"15.,778U8* >$:%#!# $1X*1.5.;.&$(R&&.8A%&1! >5A&.):8=&+,$()"5:;%.;"8%
8*= )*5+8%K;1A8*=",?:5&*:&17F8/18 ":%%8$# 81X*1.5.;.&$(P85G&! ?5:/;8* )"5:;%.;"8%4*56&"15.,778U8* >$:%#!$
;<0("3(&7&FuHF4YFEF03TFU ,<@0IXIFU ?/0W!8&$*58 %;=%$05! 3V@FYIFV/40WUIVTSI[HTFU I4 +I[FT/XLI435&IQTFF4 @3ISV/X&E,?@SI
FQLI[ITFU @/0W@0
$ 34U TLF
V@
$ 3TV@FYIFV0FZF0NFSF;%1#:i 34U %1$:$ #! SFV@FYTIZF0W5,T@/@H03TI/4 0FZF0! !
@0
N3V
8#1"$i 34U &L344/4oVI4UFQ"K
/
$ S34JFU XS/<%1%>; ! T/%18!$ "! NITL 34 3ZFS3JFZ30HF"
@/@
$ /X%1#8% 95,4J
TLFXIZF@/@H03TI/4V! @/@H03TI/4 N3VFV@FYI30W@0
="$18$i!K
/
=%18!$ "$5+L/VFSFVH0TVI4UIY3TFU
TL3TTLFSFN3V3SF03TIZF0WLIJL 0FZF0/XI4TS3V@FYIXIYJF4FTIYUIZFSVITW5,430WVIV/X0FYH03SZ3SI34YF",\h.,$
UF4VTS3TFU TL3T3LIJL UFJSFF/XJF4FTIYUIXFSF4TI3TI/4 /YYHSFU 34JTLF@/@H03TI/4V"
(
&+
=%1!#: "!!f%1%%#$5
+LF@S/@/STI/4 /XJF4FTIYZ3SI3TI/4 34J@/@H03TI/4VN3V!#1:"i! NLI0FTLF@S/@/STI/4 NITLI4 @/@H03TI/4 N3V:"1!$i5
,VI
$ /X%1!8% ; 34U TLF@S/@/STI/4 /XUIZFSVITW
34J@/@H03TI/4VNITL &L344/4oVI4UFQ3"K
V@
6K
@/@
$gK
V@
4/X%1!"# >5\34TF0TFVTVL/NFU TL3TTLFSFN3V4/VIJ4IXIY34T
Y/SF03TI/4 [FTNFF4 JF/JS3@LIY30UIVT34YF34U JF4FTIYUIVT34YF""=%1:;9 :!!=%1;%: :$ 34J@/@H03TI/4V5,UF4US/JS3<
Y/4UHYTFU [Wk?K\,
"$1#$ UF4VTS3TFU TL3TTLFS34JF/XJF4FTIYVI
HTI0I]3TI/4 /X!D%;=%$05[3VFU /4 JF4FTIYUIZFSVITWN3V@S/@/VFU5
=#> ?*".0&7!8&$*58 %;=%$05# &E,?# @/@H03TI/4# JF4FTIYUIZFSVITW# JF4FTIYUIXFSF4TI3TI/4
7!
#
# Ï
7
¼
&
E`æW*ÈöÛK;>
&E,?
[77
E`æW*
"!8&$*58 %;=%$05$
«[\(
"?3F/4I3YF3F$
[\Û
"!8&$*58$
W*î
" &FYTI/4
P$;.8*$
¤<` î
" &H[VFYTI/4 \&%868,8*8&$
§
¨
"A/4J! #;;9$!
fAqç{>vÍd4
%
E`æ
W*6[äuAq&æ
(
}&¼3
!
ÍutÕå
"$ 9%% <
j
$(
ÎÃÚΤ¥LÃ
!
=Q[0Û;
dE
"
@)¼
! $%%9$!
w%u¨
(
Î+]
(
e-®
7¡®¼
%
E`æW*#×úÚ
$* =#%"
ºS^
¼
! #;;"$!
5;¬Óq¯
"
[_Ù¼
! #;;9$!
+
`ö%
%
ìÍE`æW*gt½"
81: `E
(
×
k
!
~{*/a
!
«w¹>q\Î
!
Çy·Ut>
M]Ñ4ÑÑ
#
ìÍW*RC{E`æW*
æW*
"!8&$*58 =&%868,5Z3SD%;.&8$
b{æ×`
Ç
!
ÇÞ
!
E`æW*髦§
(
ç
(
÷æ×
(
c×W*»¬7tE`»¬>÷7¬
5f¸>«
!
E`æW*Ûdä¬
!
°uÍÃEF
8fEÒ£
!
[ä×اe
!
ÚÅ_E
!
!
Au©ªá
"
e¦¼
! $%%!$%
E`æW*>ÈöÛK;7%Èö]PQ
=u;<ªìÍFG
(
˧6»¬
(
uvW*¯°»
¬>`×¼{ĸâã
%
hE`æW*>µrd
/
!
([äçèfd
!
sghNRòW*¯¼!
Ú3í[Êöß|¬·¦
"
ij¼
! $%%8$%
ÀP
QÎHÑÒ
!
{wE`æW*>[W3í
"
ºS^
¼
! #;;"#
RQ¼
! #;;;$(
Íá)
"
Ge&¼
!
$%%9#
G¹¼
! $%#%#
§J¼
! $%#%$(
Èö)
"
&
ÉÖ¼
! $%%!$
7k%)
"
@^¼
! $%%;$
¼Ð{
Ê$"#
!
h=ª%QòÈöÛK;>PQ"
{"#
%
v w 5 ° ® Û á ;
" VFuHF4YFSF03TFU
3<@0IXIFU @/0W«Ê¬6>[Óå
DO
"-I&.8%D! $%%#$!
[«°ç¡>
#9
#"
¢}î[>
(
XÐ|¨÷U=Çî
P*,
C
lmn
(
sÓÉàÓ×Øöçm°®
!
Çn
¨¬
(
n¢úÉàÓ
(
sÓ5Îcn
y.ÍÛá;
%
æå·{Î:
(
t
(
.t
(
%;(
(
hN
(
:uMâå^ß>ç
!
¹Ð
[Ê3Þ¹uì.u
"C;:/%$&=8:.,%$5=&1$"DHU3^ &.
8%D! $%%!$(
ÁY
"C&.8 6;%8"51$ "*3J0&.8%D! $%%9$(
W*
"!8&$*58 1;(";.5:$18 $ "A34 &.8%D! $%%"3#
$%%"[$(
o¿
"P;18$ "C/HVVFX&.8%D! $%##$
¼§¨
%
&PQí¸=AqÒ×&æ´}&´>BKì
ÍFGE`æW*ö
&E,?
[!
k|*)E`
æW*%Qò
P*,
>ÈöÛK;
!
fæ¬>6
»¬Ë§7;<Ú¯¼-.[Óͨ)
%
#7
ÎHY
BABC
ÌÍ
àÈÎHfìÍE`æW*$¶>q
!
u
$%%;
$
:
%ÒAqÒ×&æ´}&´
% 9;
¢3g[«ñ
:
¢n>%
"
_
#$!
%
>J²i|
:% ^<
j
!
U=%OÍá+á
>ÛK;>ÿöp_TU
!
Æ~1%EeË
; $`8
¢3g
!
[3gÒþp^
"
3gJn!
u
: <$!
4q1
¬Ãr
(
bÉwR;Ën¹
%
f;
j<,! D! !
P! )
OÂ%
%
6
BC
·MNOP2Q
!
%
*3THS30
@/@H03TI/4V
¬G
?S/ZF434YF
K»ïÍ
&3<@0FY/UF
ÒKÚ
*H<[FS/X
V3<@0FV
,
Aq&æ´$3
?03YF
$
/X*WI4JYLI!+I[FT
" 8`% $8
D
Aq&æ´%3
?03YF
%
/X*WI4JYLI!+I[FT
8# !`! #!
Aq}&´$3
?03YF
$
/X\I0I4!+I[FT
!: >`$ #"
P
Aq}&´%3
?03YF
%
/X\I0I4!+I[FT
>8 9`# ;
)
Aq}&´&3
?03YF
&
/X\I0I4!+I[FT
9$ "`> #:
BAECSX;
RªH3$
Çî
P*,
-K
P/W0F
¼
"#;"9$
> 7
+,D
!
¹
#1:i
øqìs1DtÛNª<
!
?
ËÐW[\=ÛNìc
!P*,
K»Òu
#%%
#
-
>
+)
Ò$R
!6$% d
;Ën¹
%
BA@C5$;9
S×êë
°Õì9>Å[>
&E,?
(
XÐ|¨Â
$%
Äî[
!%%
¢|¨îÅ
!
ÃRIËÝÄõu
(
;%;((ÄõdÛ>
#>
=|¨¹u
&E,?
°®
"
|¨5%_
$$%
BAFC97$
Tã=U×3$
¹
2¡=
?E
ýÞúö÷:
!
IËÝ
$%
#
-
>ýÞúf
&
Ùv
P*,8% 4J!\J
$j
81%
<0-
6#
! U*+?V%1! <0-
6#
!
|¨Â
%1:
#
0-
6#
!>8HP*,
òÅw
$ k!
)¹
#:
#
-
Àx1
1Ũ
%
ýÞu
?+_$%%
X
?E
°®=
"
?
D(h_E,P
>
$
Rö
!
°®NK°i¼
"$%%"$& ;! d
ÃY;
8
8% V!89 d
#9
& ( )
!"
"
7
Sy
8% V!9$ d
²8
0
®Ï
# d!
"
¢KL
# ;! d
Y;
8% V!:% d
Sy
8% V!9$ d
²8
0
# 9$ d
²8
9
>i
Y;ò¾zÎ×s1Dt[
²
!
-Ê
9: O
ÄÅXÃDt
8%
!1:
#
-
.¨K
!
Dt
#1: L!
Ò¹{#
"
¤ii¼
!
$%%;$
öo×ýÞ
!
ã¡
(
Kv
%
¸°®7Dt
i
#
¢nÏÙv
P*,
>=K
%
BAHC
VÎ(¾
j
#%% [@ P*,03UUFS"
|6~Í:(D{Ñ
>
$
f[Ó<åæ
!
Ê|¨°®>DtÖ
Êé>ÄõôÏf{G;
!
ÛÊí.¨
!
O
{
"#$(
ô
"%$
n|¨°®]^
!
[
&E,?
*%!#+
_XÚ1ûP
%
¹
?h?K)*)#18$"CFL &.8%D!
#;;;$
PÅ¡£Â%Ûáí¼[
"!
@0
$(*FIoV
ÈöJ²
"\$(
Èö[:Ú
"_
&+
$%
K
K/H03ST
¼
"$%%:$
>
!
¹
&L344/4
_XÛK;ÜÚ
"K
/
$
[\¡£%s>òiÈöÛK;
"K
@/@
$
ÿÈ
öÛK;
"K
V@
$& K
/
=6
%
!
5
0/J
$
!
5
[-!
R
!!
5
f
í>_Xÿ
!-
fíÚ
# K
@/@
=#g*
%
K
/
!
K
V@
=6
%
!0/J
$
![*!
R
!*
f%Ú
!!
f
*
¢%
>Å_Xÿ
# K
@/@
gK
V@
«%sÛK;§ì
Ö»
!"K
V@
6K
@/@
$gK
V@
f%ÛK;§ìÖ»
%
¹
PB,#1#"
bvÕ¼
! $%%$$
PÅ¡£»ÑJ²
òûP
!
)Ī¤fÀ#uŹ
O(*,\h.,
")QY/XIFS&.8%D! #;;$$
PÅö[Ól[!
¡
£%s%>Yml[ä (ã¡J
²
"
(
&+
$!
)¹
\34TF0TFVT
=%>ÈöJ²3
,J²övw;ÛÈ
%
~1
*FIoV
ÈöJ²
!
=Â
%ö
k?K\,
òW[#
¹
*+&C&_
?
$1#
"E/L0X! $%%%$
PÅ¡£
23YY3SU
v¸Ú
!
=§{¢
úöòW[%
$7
]^Y[EABC5$;9+97$
Tãß(¾
¹IËÝ>
#>
¢
&E,?
|¨îÅ=
9;
SE`
æW*K»ö
P*,
°®
!
ÛNÁ
8;>
¢u(
½%>{í
!
ªRÛá;í
8:9
¢
!
Ûáí
¼[f
;%1#:i"
_
$$%
Â=|¨°®>í
Úf
#" 8`#
¢
!
°®^ßEe|
##% #` :%% [@
É
!
¢|¨i°®ÝÛá^ß
!
Ûá¼[
"":1%%i ;`>1#:i$
Ç|¨ym
%
6
EC
S×9í
$
öWdTãß
!
!
?SI
?SI
-
0
ÛáíÚ
-
@0
Ûáí¼[
!
@0
"i$
\)!g)\#"
:b_+K,K+,,,KK,_8b
:b_K,+KK+,K,,+++,K_8b
$# #" ":19#
\):g)\##
:b_+K,K+,,,KK,,K_8b
:b_K,+KK+,K,,++,_8b
$> $: ;>1#:
\)>g)\#$
:b_+K,K+,,,KK,,_8b
:b_K,+KK+,K,,+++_8b
$: $8 ;$1%%
\)#$g)\:
:b_+K,K+,,,KK,K,_8b
:b_K,+KK+,K,,++,+K_8b
$9 $: ;$1:;
\)#$g)\;
:b_+K,K+,,,KK,K,_8b
:b_K,+KK+,K,,++,K_8b
$; $: ">1$#
\)#$g)\#"
:b_+K,K+,,,KK,K,_8b
:b_K,+KK+,K,,+++,K_8b
$> $: ;>1#:
\)#8g)\9
:b_+K,K+,,,KK+,K_8b
:b_K,+KK+,K,,++,,_8b
8% $9 ;%1%%
\)#8g)\#!
:b_+K,K+,,,KK+,K_8b
:b_K,+KK+,K,,++K,_8b
$# $% ;:1$!
\)#8g)\#9
:b_+K,K+,,,KK+,K_8b
:b_K,+KK+,K,,++K+_8b
#" #9 ;!1!!
\)#8g)\#;
:b_+K,K+,,,KK+,K_8b
:b_K,+KK+,K,,+++K,_8b
$: $$ ""1%%
\)#!g)\;
:b_+K,K+,,,KK++K_8b
:b_K,+KK+,K,,++,K_8b
8# $" ;%18$
\)#!g)\#%
:b_+K,K+,,,KK++K_8b
:b_K,+KK+,K,,++,+_8b
$% #9 ":1%%
\)#!g)\#8
:b_+K,K+,,,KK++K_8b
:b_K,+KK+,K,,++++_8b
$8 $% ">1;>
\)#:g)\#%
:b_+K,K+,,,KK+K+_8b
:b_K,+KK+,K,,++,+_8b
8% $> ">1>9
\)#"g)\#$
:b_+K,K+,,,KKK+_8b
:b_K,+KK+,K,,+++_8b
$: $$ ""1%%
\)#;g)\;
:b_+K,K+,,,KK,K_8b
:b_K,+KK+,K,,+++K,_8b
#; #9 ";1!9
ÿ¡
+/T30 8;> 8:9 ;%1#:
77
"
-
0
& *H<[FS/X3<@0IXIFU 0/YI# -
@0
& *H<[FS/X@/0W@0
& ?FSYF4T3JF/X@/0WX
% +LFV3
7!
#
# Ï
7
¼
&
E`æW*ÈöÛK;>
&E,?
[EAEC
·MNOP´XEYZ[2M(¾
°_
8
½7
!
|%Qò
!
§{|¨§°®.
¨>Ûá;|n%R>[äÚn
% :
¢%
>ÛáíÚf
>% 8`$>
¢
!
%
°®ÝÛá
í
8$>
¢
!
«ªR%ÛáíÚ>
81! :`1!
]
%
n%>
!
@0
|
#:1#:i "`$18$i
É
!
ªR
%
>
!
@0
St
!
%
P
)
>d
!!
@0
¸f
%
m,mDmPm)!
òi
8#1"$i%
j]
^_`
!:
¢%R%
>ÈöÛK;St
!
ª!
!
¢%>
!
@0
dTª
%
6
@C
·MNOP\[X?E[b]W`
!
:*:G6()*&0*20$27# -,=-2?#
%
*3THS30@/@H03TI/4V
ÛáíÚ
-
@0
Ûáí¼[
!
@0
"i$
, ;9 $!1!;
D 9; #;1;:
8$> "$18$
P >" #91#9
) >% #:1#:
òi
\F34 #$> 8#1"$
77
°
&L344/4
_XÛK;ÜÚ N>E`æW*
Â%s>ÈöÛK;Ë|Êlm
"
_
!$!
YT
77
6
FC
^
54&&*&
6¡[2MV¤$E·MNOPX?dXEYZ[2M(¯
!
!
<#(?##&:*:G6()*&0*20$27# -,=-2?#2*"(4#BT :")8#"0&6>0#.
|¨
?SI
/
, D P )
K
@/@
K
V@
K
@/@
gK
V@
"K
V@
6K
@/@
$gK
V@
\)!g)\#" %1#$# $ %1%8# > %1#"; > %1%9# 8 %1%!> " %1%;$ # %1#9% : %1:!% $ %1!:; "
\):g)\## %1%$> : %1%8% " %1!!# ; %1%>" ; %1%;! : %1#8$ : %18%$ % %1!8" " %1:># $
\)>g)\#$ %1%!: $ %1%98 $ %18>9 ; %1%%" " %1%#% % %1#%# % %1#;! $ %1:$% $ %1!9; "
\)#$g)\: %1%;# % %1%>9 > %1$9! 9 %1%%% % %1$8% $ %1#8$ 9 %1$>% ; %1:%" > %1!;# !
\)#$g)\; %1%!9 % %1%$> ; %1889 $ %1%9$ 8 %1##% > %1##" " %1$>9 $ %1!!! > %1::: !
\)#$g)\#" %1#%! ; %1%9> 8 %188! > %1%:8 % %1%%: 9 %1##! ; %1$8: 8 %1!"" 8 %1:## 9
\)#8g)\9 %1%;; 9 %1%>8 ; %189$ 8 %1#$: ; %1%!; % %1#!$ $ %1$": > %1!;9 " %1:%$ $
\)#8g)\#! %1#8: $ %1#!> " %18$: % %1%8$ ; %1$$$ % %1#9$ ! %1$"; 9 %1:;: % %1!%: %
\)#8g)\#9 %1%#% # %1%%" 9 %18;; 8 %1%;% ; %1%%% % %1#%# " %1$#> " %1!9% % %1:8% !
\)#8g)\#; %1#8$ % %1%%> 8 %1$>% ; %1#%" : %1%:! % %1##$ 8 %1$># ; %1!$" ; %1:9# #
\)#!g)\; %1$:: # %1#$$ > %1!%# ! %1%9# ; %1%8$ " %1#9> " %1$9; 9 %1>8$ % %18>" %
\)#!g)\#% %1%>9 " %1%;" # %18#9 " %1%%% % %1%$" > %1#%$ : %1#98 " %1:;% % %1!#% :
\)#!g)\#8 %1##: # %1#$; ; %18>8 $ %1#>" # %1%9; $ %1#9# # %18%; 8 %1::8 $ %1!!> "
\)#:g)\#% %1%:> > %1%89 ! %1$>" 8 %1%!9 ! %1%$$ 9 %1%"> : %1#:# > %1:9% ! %1!$; >
\)#"g)\#$ %1##8 " %1%9# " %1!#; 8 %1%%" " %1%$> : %1#$" % %1$!> : %1:#; ! %1!"% >
\)#;g)\; %1#>! > %1#8" 9 %1!#% 9 %1$"# ; %1%>; $ %1$#8 % %18>9 # %1:"% 8 %1!#; 9
òi
\F34 %1#%% : %1%>; ! %18!$ " %1%98 % %1%>9 9 %1#8% 9 %1$:$ # %1:#" ! %1!"# >
77
"
K
/
& &L344/4
_XÛK;ÜÚ
&L344/4oVI4UFQ/X@LF4/TW@IYUIZFSVITW# K
@/@
&
%s>òiÈöÛK;
\F34 NITLI4_@/@H03TI/4 JF4FTIY
UIZFSVITWX/S30@/@H03TI/4V# K
V@
&
ÿÈöÛK;
KF4FTIYUIZFSVITW/XV@FYIFV# K
@/@
gK
V@
&
%sÈöÛK;§ìÖ
?S/@/STI/4 /XUIZFSVITWNITLI4
@/@H03TI/4V# "K
V@
6K
@/@
$gK
V@
&
%ÈöÛK;§ìÖ
?S/@/STI/4 /XUIZFSVITW[FTNFF4 @/@H03TI/4V5
f
%1%>; ! `%18!$ "!
òif
%1#8% 9# :
¢%
>ÈöÛK;çf%
m,mPmDm)!
óYÂ
%>
!
@0
Eeç&Êé
%
n|¨ÛNÁ>
ÈöÛK;Ee1{lm
!
ªR|¨=
\)#;g)\;
ÛN>ÈöÛK;St
!
òi
%1$#8 %!
y|¨=
\)#:g)\#%
ÛN>S
!
òi
%1%"> :%
Ã_
!
é½äÝ
!
E`æW*%s%
>ÈöÛK;||¨YàdE
!
3`Â|¨°®
Ý>nÛáí=ÈöÛK;>µ«n>
%
Â%s>ÈöÛK;
"K
@/@
$
òif
%1#8% 9!
ÿ>
ÈöÛK;
"K
V@
$
òif
%1$:$ #!
%>ÈöY
mìÿ>ÈöYm>
!"1#>i!
%s>ÈöYm
êì
:#1"!i%
½%
!
E`æW*%Ë|dt
Qò>Èö[:
%
EA@C
·MNOPXEYZ
,\h.,
[\oOÿ>ÈöYmR{
!#1:"i
Ë|u%
!
{
:"1!$i
Ë|u%s
"
_
:$!
(
&+
f
%1!#: ""!f%1%%#$!
_`E`æW*%
>ÈöYm=ÿ>ÈöYmµdE
!
%[:
dop
%
=uv>
&E,?
Ú1
!
°
?h?K)*)
¡£
>%>Ç[:Ú
_
&+
f
%1!8% ;!
¢Y
&L344/4
_XÛK;ÜÚÇÝ>%ÈöÛK;
§ìÖ»
3"K
V@
6K
@/@
$gK
V@
4
f
!"1#>i!
1oOß
%[:Qòdt
%
89
& ( )
!"
"
7
6
HC
·MNOP\[XE(p_(¾
!
!
YmñG
&/HSYF/XZ3SI3TI/4
°c
UX
ÿl
&&P
Ymî[
.3SI34YFY/<@/4F4TV
ìÿYm¼[Ö
?FSYF4T3JF/XT/T30Z3SI34YF"i$
!
%
,4J@/@H03TI/4V ! # %#!1!9 #:1%! !#1:" f%1%%#
%s
OITLI4 @/@H03TI/4V 9! # :>81#% $#1#$ :"1!$ f%1%%#
77
"
UX& PFJSFFV/XXSFFU/<# &&P& &HEAFC
·MNOPXE#U
E`æW*Â%
*FIoV
ÈöJ²>Y:ç
èf
%1%#% > %`1#>9 >!
j%
)
É>È
öJ²fSE
!
%
,
D
>ÈöJ²Se
"
_
>$% \34TF0
ÛÈ]^_`E`æW*%>È
öJ²3,J²Évwnop
""=%1:;9 :!
!=%1;%: :$%
~1
*FIoV
ÈöJ²=Â%ö
k?K\,
òW½7
"
ß
#$!
ÛAq&æ´>%
,
D
ò|Ê}
!
yÛ}&´>
8
¢%n#
ò|Ê}
!
©ª«%
)
YªR
!
¢%É
>ÈöJ²optuªR
!
¢%>Èö
J²
%
6
TC
·MNOPXEYZ`ad3|`a
!
!
%
*3THS30
@/@H03TI/4V
, D P )
, :$1:"7 #:#1!!7 >!1;87 ##$1;8
D %1%#% > #>;1:% #%#1:" #>81%:
%1%:8 9 %1%:: : 9"1%; :#1$>
P %1%89 : %1%89 % %1%>! " ::1!9
) %1#>% ! %1#>9 # %1#>9 > %1#89 9
77
"=¼sjÚ1f3,J²
" ^<$!
=¼sjXÚ1f
*FIoV
ÈöJ²
% KF/JS3@LIY30UIVT34YF" ^<$ "3[/ZFUI3J/430$ 34U *FIoV
JF4FTIYUIVT34YF"[F0/NUI3J/430$5
ß
#7
E`æW*%
*FIoV
ÈöJ²>
k?K\,
òW
BIJ5#7k?K\,UF4US/JS3[3VFU /4 *FIoVJF4FTIYUIVT34YF
EAHC
·MNOPX?ÀÁEb)(¾
fß~E`æW*%s¢ú>YmTc
!
=
§{>3göß
k?K\,
òW
"
ß
$$%
òW]^
_`
&
§{¢ú>Èöv¸ÚYT|
%1!9 `
%1;;!
Ê%>¢ú)n#âò|Ê}
%
E²[ñGuÊ%>¢ú_rÝdfb>ä
îw
!
o
,! D! P! )!
¢%s>¢úv¸
Úidt
#
h1{Ê$%>¢ú$#ò|Ê}
!
oÈöv¸Úf
%1>%
!
%>
:
¢3g
"
K
»
!"! :#! :"! :;! >#$
òfʶ
!
æ%>ª!¢
úYªR%>¢úòfʶ
%
87
]/Y)/
@ABC
·MNOPEYZ[2M
ÊZy8
!
[äçèfd>ç{§¨¬>Ûá
;QòÖ²ä¬
"
¤}¼
! $%%>$!
hªñn!P
Qµr$ç{©ª¨¬f;dt>YmQ
ò
!
oR?_Ò
"F5"5$=&*="$* :/5*&*1&$ "
î*¼
!
$%%>$
¼
%
&PQRE`æW*¬Qò>Ûáí
¼[t"
;%i
j
!
tuÊ î>~W*
"!8&$*58 =&%868,5$"9;18#i$"
G¨"¼
! $%%:$!
_
`E`æW*·{dtQò>ÈöÛK;
%
|§P
Q>
:
¢%R
!
%
>ÈöÛK;St
"!
@0
=
"$18$i!&L344/4
_XÛK;ÜÚ N>ÛK;f
%18!$ "$!
ìÎTUµræ%Au;Ëdâ(>
>{ÍÃÄÅX
!
½#ÇÞ\ßuv>ÈöÛK
;
"
@)¼
! $%%9$#
ªR%Ç6[äu&î>
¸Íd4&¨¡Þÿ3
!
8f
(
Ìàÿq
!
Í
ÃEF
!
ÏÉ=ªYc½£
!
@¢úÚeE
!
7
éÈöÛK;E!
%
@AEC
·MNOPXEYZ(K
\FF^I4V
¼
"$%%#$
@"#Y
&L344/4
_XÛK
;ÜÚ=o;ån#µTNc&Åú7é>ïl
ná`
%
|&PQR
!
Þ¹
,\h.,
[\WÝ>
(
&+
"%1!#: "$
Y
_
&+
"%1!8% ;$
Tª
!
¿¥¯e
u¢Y
&L344/4
_XÛK;ÜÚ¡£WÝ>%
ÈöYm
"%1!"# >$!
hòi¿OßE`æW*%
Ë|dtQò>Èö[:
!
óf%Ë÷-
.ß½#
%
%Èö]ÿÿÀ½ÛÇÈ>?@
!
ðñÍÃ^ó:
(
ÇK
(
q§ã
(
¬Ó±Ò
¼
"KF&.8%D! $%%:# BIVYLFS&.8%D! #;;"$%
&PQ
R
!
7éE`æW*%Èö[:dE>>ǽ
#Y¬Ó
(
`%>ö{w
%
E`æW*5;¬
Óq¯
!
¬ÓdE
!
[Qµ¥½"
#18 Y<"A/4J&.
8%D! $%%#$!
ö
!
y(Û+`ö%
!
ª`%±
!9
7!
#
# Ï
7
¼
&
E`æW*ÈöÛK;>
&E,?
[ß
$7
E`æW*§{¢ú>
k?K\,
òW
BIJ5$7+LFk?K\,UF4US/JS3
#% ?/@H03TI/4 Y/UFV3SFTLFV3
!
ó$Ç
Ƚ#ÑÒß%>ÇK
%
Ê$%¢úd
!
!
ÇÍÃ^ß:N
!
hµÍÈöY
!
Ãy7é
%sÈöYmP
!
%Èö[:Ï
!
o%
)
½#µÍßÈöY
!
%òWßYªR
!
¢%É>ÈöJ²optuªR
!
¢%>
ÈöJ²
%
ÍÃ>niÊ;ÿÿ«%s¢úlm¨%
lm>µT>Ç
"
/
! #;;!$%
Aqt>â}
É
!
°ut>·¯à>lm;!f^jñ>Ö
#YÖ#GÜÉ2ã?@
!
+[ßt>Í[ç
>%&>LÃ
"
/Ía
! #;"%$%
¢úòWß
EÛÚ%s¢ú_rÝdfb>äîwo%
D!P!)#
h1{Ê$%>¢ú$#ò|Ê}
!
o
%
!
(æ%>²[K»
"
ïÍ
!"!:#!:"!
:;!>#$
|=|¨=ªö
P*,
°®i#.Í
Û¢çm;Äõ
!
óÃ
P*,
Qòýßó$¢ú
Y%sªR¢úË|dE>lm
%
WXâ>
«
!
YªR%vÖ
!
%
>[ä3²Ög
Q
dª
!
ÇÞ-[
%s¢úlm7
%Yª
R%lm>ÇȽ#YQÇÓ{w
!
hó^¸
öÊ_>PQ
%
@A@C
cdH¢£ef
¤¥¢Y
$
$>EÙìÎTU
!
êµrß
:
¢dE>E`æW*%
3
Éñ{u"#+=3
×k[ä{
#
¢%
"
o
! $%%;$4!
ó
:
¢%
:9
& ( )
!"
"
7
[äçèv2fd
!
E²[%[äu8WÌàÿ
q>3
!
y(çg+
(
×
(
`×
(
`Ú
(
¸
(
9
(
¬Ó¼_XñäiY:nE
!
&PQÞ
¹
&E,?
åé¿OÝß%Ë|dE>Èö[
:
!
ó$3`ªìͬê½Ä©ª>^EAµ
!
Ç
Þ
!
W^¸ÒÅ{>ÓÔ;
%
ªÊ
!
|[ä3;<×
!
=Èö
ÛK;t>%öÆ3;<
!
o&PQRÈöÛ
K;dt>%
#
ª¿
!
¸Æ¬
þ
!
¬
®=E`æW
*>ÈöYmç
!
®Ïþ>%Ú%
s>Ú
!
;2Ö3;<>{;
#
ªr
!
¢Y
ÇÍa¦§¨î¦PÕ¥ÄÇÿde>¢ú
ö°qÉy
!
½#«;
%
jM]få¶E`æW*>
¦§
!
*)ª|¯&R>Þ¹
!
kÄ=ª;<}
Á¸>Lह
!
E`æW*uv>ÈöÛK;
f»¬Ë§-.߬<;2
%
X Y + ,
[_Ù
!
ºS^
!
/(
5#;;95
RìÍW*q¯ç;PQ
5
¯°)"
! $!"$$& #"% 6#"!5
/
5#;;!5
%8
|§¨[W)R>Þ¹$/
!
G
7
Ó
5
§¨ö:ͨ)
5
&
E)Ýv
! 8>: 689!5
/Ía
5#;"%5
Aq&æ¨36Ír/#>LÃ
5
qóà¨
Yq8W
! #""#$& :$ 6:"5
i
7
j
!
ãæ
!
GøÙ
5$%%85
~W*>ÛK;rÐ
5
AB
§¨)"
! $8"$$& $#" 6$$85
°
7
i
!
îÛ
!
xk
!
¼
5$%%"5
Rí»[\W*³î&2¬
>µr7
5
¯°)"
! 8:">$& ":8 6":"5
RQ
!
ãx
5#;;;5
[\ÛW*î>[WG¢[WA,
5
§
¨[W)"
! 89 "!$& 8:# 68>"5
ºS^
!
/(
!
u
7
À
!
¼
5#;;"5
E`æW*[W)3í>P
Q
5
§¨PQ
! #""!$& #:$ 6#::5
&ÉÖ
!
?ø
!
l
7
!
¼
5$%%!5
u
,UL
Ç5>W*
î
"&FYT5P$;.8* P5$
¬w
5
¯°)"
! 8#":$& >$9 6>8$5
î
7
*
!
ËÍ
!
Göò
5$%%>5
R?_ÒÈöÛK;PQ
5
&
()
! !#"#$& :9 6>!5
o
5$%%;5
ÏW*
(
æW*
(
E`æW*|¬YÖ3;
B&E)B¯)í/u
5
§J
!
îÍ
!
)?
!
¼
5$%#%5
©ª§¨E`æW*YÍÃ3
´çè>w
5
&()PQ
! $8"!$& !"9 6!;$5
e
7
¦
!
~
7
F
5$%%!5
R¨¬í×
5
B
&
t¼±§Ýv
! 8$85
¤
7
}
!
§Þ=
!
»{%
!
¼
5$%%>5
x¹
E,?P
å[\©ª§
¨W³-¬>Èö]
5
Íá)"
! $>"9$& $8#8 6$8$#5
¤ii
!
@(
5$%%;5
x¹
2¡÷:W*
&E,?_?E
ýÞú
5
[Ó§¨§¬
! 9"#$& #;; 6$%85
@
7
)
!
bɹ
!
¸
7
<
5$%%95
AqE`æW*ÍÃ8(p_T
U
5
¿§C()
! "!$& $:% 6$:85
Ge&
!
¤[t
!
YeR
!
¼
5$%%95
©ª§¨E`æW*
"!8&$*58
%;=%$05$
¬Úàá
5
Íá)"
! $9"8$& #$!$ 6#$!95
G¨"
!
郎
!
i
7
j
!
¼
5$%%:5
~W*ÈöÛK;>
(&&E
[
5
ͨÛK;
! #8"$$& #%: 6###5
G
7
¹
!
7
p
5$%#%5
E`æW*´k¸¬>ÍáíPQ
5
¿§C()
! "#$& 8#! 68#"5
bvÕ
!
¢
7
5$%%$5
úÈö)PQR>Ú1A,
$
5
E,?P
Ú1>
,\h.,
[5
ͨÛK;
! #% "!$& !8" 6!!!5
@^
!
r
!
º
7
i
!
¼
5$%%;5
²[W*`%$X¼]>P
Qp"
5
fgCE))"
! $9"!$& !>> 6!9%5
DHU3^ A! &LF3S<34 E!?3S<3^VI](5$%%!5\/0FYH03SYL3S3YTFSI]3TI/4
/X [HX30/JS3VV JFS<@03V< HVI4J VFuHF4YF_SF03TFU 3<@0IXIFU
@/0W8$" 688!5
P/W0F22! P/W0F2-5#;"95,S3@IU P*,IV/03TI/4 @S/YFUHSFX/SV<30
uH34TITIFV/XXSFVL 0F3XTIVVHF5 ?LWT/YLF
)QY/XIFS-! &Z3SI34YFI4XFSFU XS/<
BIVYLFS\! \3TLIFVP5#;;"5E,?PZ3SI3TI/4 I4 SF03TI/4 T/@/@H03TI/4
VI]F 34U @034T XIT4FVV I4 TLF S3SF _&*.58*&%8 &"+8*5:8
"KF4TI343YF3F$5,
KFGHFtH4! RL34J-I4[I4J! CH34 C/4J
"KF4TI343YF3F$ UFTFYTFU [WI4TFS_VI<@0FVFuHF4YFSF@F3TV"(&&E$5
DI/UIZFSVITW34U /4VFSZ3TI/4! #!"!$& "!; 6">#5
K/H03ST\B! EI[FIS/& ?! -/Z3T/\D5$%%:5KF4FTIY! 34U V@3TI30YL3S3YTFSI]3TI/4 /XTN/@/@H03TI/4V/XP8A&8 (51.;%5(&"8
\3ST5")H@L/S[I3YF3F$! I4 UIXFSF4TVHYYFVVI/430VT3JFV5DS3]I0I34
,SYLIZFV/XDI/0/JW34U +FYL4/0/JW! !""$$& $9: 6$"!5
A34 GI3/W34! O34J -I34JVLF4J! -IH RLF4Jo34! &.8%5 $%%"35
L3S3YTFSI]3TI/4 /XVFuHF4YF_SF03TFU 3<@0IXIFU @/0W3430WVIV/XTSFF@F/4W[HU V@/STV5&YIF4TI3A/STIYH0THS3F! ##:"8$&
$># 6$>95
A34 GI3/W34! O34J-I34JVLF4J! &LH MI4JW34! &.8%5$%%"[5\/0FYH03S
YL3S3YTFSI]3TI/4 /XTSFF@F/4WJFS<@03V< HVI4J&FuHF4YF_EF03TFU
,<@0IXIFU ?/0W"8g!$&
#>$ 6#9;5
A/4JPFWH345#;;95!8&$*58 " ?3F/4I3YF3F$ I4 GI]34J"+I[FT$5
*/Z/4! 9"$$& #:> 6#>#5
A/4JPFWH34! ?34 r3IWH! *IYL/03V2+5$%%#5?3F/4I3YF3F
$
OH RC!
E3ZF4 ?A5B0/S3/XLI43& ./05>5DFItI4J& &YIF4YF?SFVV34U
\IVV/HSID/T34IYK3SUF4 ?SFVV! #$9 6#8$5
-IK! MHIS/VB5$%%#5&FuHF4YF_SF03TFU 3<@0IXIFU @/0W"&E,?$! 34FN<3S^FSVWVTF<[3VFU /4 3VI<@0F?ESF3YTI/4&(TV
3@@0IY3TI/4 T/<3@@I4J34U JF4FT3JJI4JI4 C"815:85+LF/SFTIY3034U
,@@0IFU KF4FTIYV! #%8"$g8$& !:: 6!>#5
\FF^I4V2B! D303SU A)! \YY3STLWD5$%%#5KF4FTIYZ3SI3TI/4 34U
0FYH03S[I/JF/JS3@LW/X3*/STL ,
&YIF4YFV! #>$"#$& #># 6#>;5
*3J0*! OFI034U 2! -FNF0F4 E5$%%95PFTFYTI/4 /XP*,@/0WI4 VHJ3S[FFT[H0^V[W&E,?34U E,?P <3S^FSV52/HS430/X
DI/TFYL4/0/JW! #8#"$$& &8$ 6&8:5
E/L0XB25$%%%5*+&C&_@Y& *H
CFL B!C34JE!D/W0F+5#;;;5?h?K)*)ZFSVI/4 #18$!
343U3& k4IZFSVITW/X,0[FST35
C/HVVFX\! 23
!9"8$& #"; 6#;;5
!
!"#$
7
_
7
@
"
>9