为揭示文冠果体胚形态建成过程中蛋白质组分的变化规律和确定标记性蛋白,以文冠果种胚离体培养诱导体胚发生过程中所获得的非胚性愈伤组织、胚性愈伤组织、球形胚、鱼雷胚、子叶胚及再生植株为材料,采用双向电泳进行文冠果体胚形态建成过程中蛋白质组分变化的研究。结果表明:非胚性愈伤组织的蛋白质组分最少,随着体胚形态的建成,蛋白质组分逐渐增多,再生植株时期的蛋白质组分减少。胚性愈伤组织、鱼雷胚、子叶胚及再生植株的标记蛋白质依次为23.0 ku(pI 6.9)的蛋白质、27.1 ku(pI 7.5)的蛋白质、25.1 ku(pI 6.6)和26.2 ku(pI 6.6)的蛋白质、23.2 ku(pI 9.5)的蛋白质。
In order to investigate the changes and the characteristics of proteins during somatic embryogenesis of Xanthoceras sorbifolia, two-dimensional electrophoresis was used to study the changes of protein expression at different development stages of somatic embryogenesis, such as non-embryogenic callus (NEC), embryogenic callus (EC), globular embryoid, torpedo embryoid, cotyledonary embryoid and regenerated plant which were obtained from seed embryos of X. sorbifolia. The results showed that the protein components were the fewest in NEC, and increased gradually during somatic embryogenesis. The protein components decreased when the plant was being regenerated. Particular proteins spots of pI/MW (molecular weight) were found at the different development stages. Particular proteins spots of pI / MW could be the characteristic protein for the particular stage. Protein spot of 23.0 ku (pI 6.9) was observed at the EC developing stage. Protein spot of 27.1 ku (pI 7.5) was observed at the torpedo embryoid developing stage. Protein spot of 25.1 ku (pI 6.6) and 26.2 ku (pI 6.6) were observed at the cotyledonary embryoid developing stage. Protein spot of 23.2 ku (pI 9.5) was observed at the regenerated plant developing stage.
全 文 :!!"
" !
#
#
# $ % %
$
#
%
& ( )
&()*+(, &(-.,) &(*(,)
./01!"!*/1#
2345!# $ % %
[Y \Ǩ&±UGö<È}ä°
"
ijk
%!#
6
8lc
%
6
fmn
%
6
3
6
o
%
6
Epq
%
6
)rF
7
"%1
ï&M)N9()V%G)A
6
ï
%$$$C7# #1
*M)NO())A
6
TÒ
$"%$$$#
71
ï(%É)A
6
ï
%$##$9$
!
6
"
!6
{()[Y \Ǩ&±UGöÈ}ä°<²½âoKÒ|È}
!
#[Y ~Çq\ÏW
Êu\ÇcNUGön<ÇË`°Ì
&
ÇË`°Ì
&
é¨Ç
&
Í-Ç
&
%<Çò4N8n{o$
!
>X¶
omÎ[Y \Ǩ&±UGöÈ}ä°²½
¥¦
ÇË`°Ì<È}ä°$Â
!
æç\Ǩ&<±
!
È}ä°ª«¬
!
4N8nÓ#<È}ä°·Â
%
ÇË`°Ì
&
Í-Ç
&
%<Çò
4N8n<|È}äst{
#71$ bO"Z(918$
<È}ä
"1% bO" Z("1B$
<È}ä
B1% bO" Z(919$
#91# bO
"Z(919$
<È}ä
G# bO"Z(81B$
<È}ä
%
#$%
6
[Y
#
\üÏÇ
#
È}ä
#
¶omÎ
&()*
! &"%C1!7&h8!9666
+,-./
!,666
+01*
!%$$% F"!CC"#$%%#$# F$$B7 F$B
ÓÔÕ#
#$$8 F%# F%!#
Ö×Õ#
#$%$ F$# F##%
ØÙÚÛ
BÜÝÞ()ØÙ
"7$$"$9%#$ #
*M);)ØÙ
"#$$B%$$C$ %
"H!Ð{*Ѫ/
%
N2*+2%+/+/0*%#+/!MF*#&&+/@)*+/5 ;,2%+$!,7*>5#/#&+&"3%)4.#$(%**(,"5/"%
TO mOQ/@P
%!#
6T=/&QO:M@
%
6-M23@P0=@
%
6GO hM@P
%
6JO KM@PXQO=@
%
6S=@Pm=@QO=
7
"%12*;#1#*+?/*;*1/:%;B:/#):#8%)- ?/*#:3)*;*19! ?#/M/)1 N*&96)/7#&8/96?#/M/)1 %$$$C7#
#12*;#1#*+,/+#B:/#):#8! G#=#/A1&/:4;4&%;6)/7#&8/96?%*-/)1 $"%$$$#
716)/7#&8/9+*&B:/#):#QE#:3)*;*19*+?#/M/)16?#/M/)1 %$##$9$
-7&%*2$%6(@ />Y3>U/M@W3?UMP=U3UQ3XQ=@P3?=@Y UQ3XQ=>=XU3>M?UMX?/VZ>/U3M@?YO>M@P?/:=UMX3:4>R/P3@3?M?/V
W%)3*:#&%88*&=/+*;/%! U\/IYM:3@?M/@=0303XU>/ZQ/>3?M?\=?O?3Y U/?UOYRUQ3XQ=@P3?/VZ>/U3M@ 3^Z>3??M/@ =UYMV3>3@U
Y3W30/Z:3@U?U=P3?/V?/:=UMX3:4>R/P3@3?M?! ?OXQ =?@/@I3:4>R/P3@MXX=0O?"*)$ ! 3:4>R/P3@MXX=0O?")$ !
P0/4O0=>3:4>R/MY! U/>Z3Y/3:4>R/MY! X/UR03Y/@=>R3:4>R/MY =@Y >3P3@3>=U3Y Z0=@U\QMXQ \3>3/4U=M@3Y V>/: ?33Y
3:4>R/?/VWF8*&=/+*;/%5+Q3>3?O0U??Q/\3Y UQ=UUQ3Z>/U3M@ X/:Z/@3@U?\3>3UQ3V3\3?UM@ *)! =@Y M@X>3=?3Y P>=YO=0R
YO>M@P?/:=UMX3:4>R/P3@3?M?5+Q3Z>/U3M@ X/:Z/@3@U?Y3X>3=?3Y \Q3@ UQ3Z0=@U\=?43M@P>3P3@3>=U3Y5<=>UMXO0=>
Z>/U3M@??Z/U?/VZ(e[S ":/03XO0=>\3MPQU$ \3>3V/O@Y =UUQ3YMV3>3@UY3W30/Z:3@U?U=P3?5<=>UMXO0=>Z>/U3M@??Z/U?/VZ(
e[S X/O0Y 43UQ3XQ=>=XU3>M?UMXZ>/U3M@ V/>UQ3Z=>UMXO0=>?U=P35<>/U3M@ ?Z/U/V#71$ bO " Z(918$ \=?/4?3>W3Y =UUQ3
)Y3W30/ZM@P?U=P35<>/U3M@ ?Z/U/V#"1% bO " Z("1B$ \=?/4?3>W3Y =UUQ3U/>Z3Y/3:4>R/MY Y3W30/ZM@P?U=P35<>/U3M@
?Z/U/V#B1% bO "Z(919$ =@Y #91# bO " Z(919$ \3>3/4?3>W3Y =UUQ3X/UR03Y/@=>R3:4>R/MY Y3W30/ZM@P?U=P35<>/U3M@
?Z/U/V#75# bO "Z(81B$ \=?/4?3>W3Y =UUQ3>3P3@3>=U3Y Z0=@UY3W30/ZM@P?U=P35
=#> ?*@&6W%)3*:#&%88*&=/+*;/%# ?/:=UMX3:4>R/P3@3?M?# Z>/U3M@# #IJ303XU>/ZQ/>3?M?
66
[Y
"W%)3*:#&%88*&=/+*;/% $
{3Æ%(
"&=ZM@Y=X3=3$
[Y ô89
!
;<íëÁêë
!
,
¦B9Ð<ëS[$ÑX89
!
ÙNNÒ[
&
ÓX
[å¢Ô[
%
[Y EÈ{
)
àá
*
#QBÜ
P\N9Õ[k~
!
~S[§{
BBD d9"D "
C=
!#$$"$!
~SÐ<
%%
~[Ö×<Ø
fQÞöb
%9
d
%C
±Q
!
ÕNNN9Ò[
"
Ù
Qö
!#$$" $!
[Y ÓX[ö< [,öÑ
)
tÏÚ
*
{¡XÓX[
&
N9Ò[
òÑ9±NN.[Y ~%ë
S~È°p¿0[Y \ÇcN<ÏW\H
"
Kâú
!#$$!$!
YZcÀ
=
# j
V[Y \Çc
N8Ï£¤
"
Kâú
!#$$C$%
\Ç<¨&±nØôiy
!
IÈ}ä,Øô
¥
Û h
!
j¤8 â Û
"]#% (%98$
"2>=@_#%;5!%8C8$&
"Ü
"P&95% 8%/7% $ "SPM>/M^#
%;5!%889 $&
@²³
""%4:48:%&*% $ "l:3Y=#%;5!
%88"#
4Ý
! %88" $&
Þö
" B%)%;4( %;=4($
& ( )
!"
"
6
"A)/;#%;5!#$$$$
89\ÇÈ}ä°<;om
ÎM
A¦È}ä°V\Ǩ&±8
Ï£¤
%
ÌФ\Ǩ&±UG<
9
vcÛÓ#
È}ä°<¶omÎ
S
YZ#[Y ~Çq\ÏWÊun<ÇË`
°Ì
&
ÇË`°Ì
&
é¨Ç
&
Í-Ç
&
%<Ç
&
4N
8n{o$
!
X¶omÎ%G
!
.[Y \Ǩ&
±UGöLÓ#<È}ä°YZ
!
ßb(
)ÃÈ}ä°<²½âo
!
à|È}
!
{Ñ
¿¼½ÞV\ÇcÛ£¤<|È}7V
ã¤<Øô
&
=RTiy[Y \Ç
!
I{WX[Y \ÇcN\Ha
;~i°òá
!
Ï{[Y Õ%Ç<¨&±
%
%6
o$VrP
ABAC
ö
[Y <ÇË`°Ì
&
ÇË`°Ì
&
é¨
Ç
&
Í-Ç
&
%<Ç
&
4N8n
%
ABDC
`Ñ
%1#1%6
;,-
6
[Y <±Z~
Çb
L
B
j#!!IJ% :P,-
F%
j
âµ
#$ P,-
F%
j
ãÖ
9 P,-
F%
<ÏWØmÊuNNÇË`°Ì
!
b
L
B
j#!!IJ$1B :P,-
F%
j
âµ
#$ P,-
F%
j
ãÖ
9 P,-
F%
<ÏWØmäxÏWnÇË`°Ì
!
b
L
B
j9IL,% :P,-
F%
j*,,$1#B :P,-
F%
j
âµ
#$ P,-
F%
<ÏWØö
!
NNé¨Ç
&
e¨Ç
&
Í-Ç
&
%<Ç
!
%<Çb
L
B
j9IL,$1B :P,-
F%
j*,,
$1B :P,-
F%
j
âµ
#$ P,-
F%
<ÏWØöcÛ±Ð
d<<4N8n
%
%1#1#6
;
6
uTb
FC$ c
$1% P<.åæöYç±»
%
»°dèÐÈ}ru
,
å
"
S
%$D +,!
$1$"D
,
F
éØêë<ìí´å
$
<
B$ :-
qeÐ
ö
!
îº
! F#$ c
X;h
# Q!
#Qîï
%
t
% ! c
X
!%# $$$ >,:M@
F%
qe
7$ :M@%
ðm+å
!
zdÈ
}ru
L
å
"
S
$1$"D
,
F
éØêë<ìí´å
$!
Xñòóôõ
! F#$ c
X;h
# Q!
#Qîï
%
t
%
! c
X
! %# $$$ >,:M@
F%
qe
7$ :M@%
Ppru
7
t
!
+¼È}äö÷9{¼}´
! F#$ c
ø2Ù
ðXÁb
F#$ c
T!X
%
JKu
$1$7 P
È}
»;d
%1B :-
qeÐö
!
d
7$$
-
k;?
å
"
S
8 :/0,-
F%
ùõ
& !DG,<&& #D:=0RU3
7I%$!
¶ú"Ò&
$!79 d7" c
X"û
% Q% ! c
X
!
%! $$$ >,:M@
F%
qe
%B :M@!
um+å
! FC$ c
XT
! X
%
¶ o m Î X < U í ,
,:3>?Q=:
LM/?3XM3@X3?
õ|
MV¿ú
!
%
omÎ
"
mü-mÎ
$&
¨^ýe
&
!
#
omÎ
" &J&I<,T)$&
Mþ P þ ´
&
X
l[,N \3>0//b
#%$$N-
*YýÿDU!^ÿD
"9$$ 4ZM!
<
´
$!
ÞeX9WÕý"³w
£,cÛÅÔ<È}\
"
Kâú
!#$$B$!
¿¥¦¡¬<
&
·Â<
&
Ñ¿À<
&
c<
È}\òLÓ#9Ð<|È}ä
%
Pp
#
t
%
#6
V
DBAC
+H·:ICJ
[Y ~Ç[b
L
B
j#!!IJ% :P,-
F%
j
âµ
#$ P,-
F%
j
ãÖ
9 P,-
F%
<®\ÏWØm
!
Ï»:N
N!¦#í´
&
äT$9
&
ý
&
¥y«Ô<
ÇË`°Ì
"
%I%$#
ÇË`°Ìb
L
B
j
#!!IJ$1B :P,-
F%
j
âµ
#$ P,-
F%
j
ãÖ
9 P,-
F%
<®\ÏWØm
!
fs½
!
cÛ±#íí´
&
ä
T$9
&
ý
&
¥yÍ«Ô<ÇË`°Ì
"
%I#$#
ÇË`°Ìb
L
B
j9IL,% :P,-
F%
j*,,
$1#B :P,-
F%
j
âµ
#$ P,-
F%
<å\ÏWØö
!
äx
½
!
stNN¥y«Ô<é¨Ç
"
%I7$&
ÐÐ
#
v%<_Ø
%¨<Í-Ç
"
%I!$
ÐÐ
#
d%<<%<Ç
"
%IB$#
%<Çb
L
B
j
9IL, $1B :P,-
F%
j*,, $1B :P,-
F%
j
â µ
#$ P,-
F%
<ÏWØöcÛ±Ðd<<4N8
n
"
%I9$%
DBDC
+H·:IKLhMN&COPQg(
[Y ÇË`°Ì<È}ä°,
%I"
)
!
È}ä°ÃYcÛÓ#ÁÂ
!
Ãö
#$1% bO"Z(71%$
<È}ä
%C18 bO" Z(71%$
<È}
äbÇË`°Ìöc
!
bé¨Ç
&
Í-Çö¥P
Ñ¿À
!
M¥æ´
!
b%<Ç4N8nö¥
c
# #B1$ bO" Z(C1C$
<È}ä
#91$ bO" Z(C1C$
<
È}ä
!
bÇË`°Ìö¥$M
!
é¨ÇÓ#
·(
!
Í-ÇÓ##ec
!
ÙaVÇ
%
[Y ÇË`°Ì<È}ä°,
%IC
)
!
VÇË`°Ì£
!"
~È}ä2
%91% bO
"Z(!1#$!%918 bO"Z(!17$!%!1C bO" Z(!1B$!%91B bO
"Z(!19$!%!18 bO"Z(B1$$!#$1$ bO" Z(B1"$!#81B bO
"Z("1$$
<È}䥦¡¬M
!%B
~È}ä2
%917 bO"Z(71%$!%91B bO"Z(71%$!%91C bO" Z(71%$!
%"17 bO " Z(71% $! %"1B bO " Z(71% $! 7$1$ bO
"Z(71% $! %"1B bO " Z(71# $! 7"1B bO " Z(71B $!
!B
6!
#
# Kâú
[Y \Ǩ&±UGö<È}ä°
7"1B bO"Z(719$!7"19 bO"Z(719$!7717 bO" Z(71"$!
7!1$ bO"Z(71"$!!%1" bO"Z(!1#$!%!1B bO" Z(!17$!
%"17 bO"Z(B1#$
<È}䥦¡·´
!
?ÓÑ
¿À0
%"
~È}ä
!
2
BB1$ bO " Z(!18 $!9719 bO
"Z(!18$!B$1# bO" Z(B1#$!971! bO " Z(B1#$!9#1#
bO"Z(B17$!B$1# bO" Z(B1B$!B71$ bO" Z(B1B$!%"1"
bO"Z(91C$!%"1" bO"Z(918$!#71$ bO" Z(918$!!%1"
bO"Z(C1$$!!%1" bO" Z(C1#$!#71B bO" Z(C1B$!#!1$
bO"Z(C1B$!#91# bO"Z(C1B$!!$1! bO " Z(C1"$!!%1$
bO"Z(C18$
<È}ä
!
Ãö
#71$ bO"Z(918$
<È}ä
{ÇË`°Ì9Ð
!
,ÇË`°Ì<|
È}
%
[Y é¨ÇÓ#<È}ä°,
%I8
)
!
VÇË`°Ì£
!%%
~È}ä2
%81# bO
"Z(91%$!7%1B bO"Z("17$!#%1% bO" Z(C1B$!#%1B bO
"Z(C1"$!%81$ bO " Z(817 $!%81% bO " Z(817 $!%81B
bO"Z(817$!%81" bO" Z(81B$!%"17 bO" Z(819$!%81"
bO"Z(819$!%81" bO"Z(81"$
<È}䥦¡¬
M
!!
~È}ä2
%91C bO"Z(!18$!%"17 bO"Z(!18$!
#71% bO"Z(B19$!#!1$ bO"Z(B19$
<È}䥦
¡·´
!#71$ bO"Z(918$
<È}äc
!
?ÓÑ¿À
0
%#
~ È } ä
!
2
%C18 bO " Z(71% $! #$1% bO
"Z(71%$!%917 bO"Z(B1"$!7#19 bO" Z("1B$!!$1! bO
"Z("1C$!7#1$ bO " Z(C1# $!7%1# bO " Z(C1! $!7#1$
bO"Z(C1"$!7%1# bO" Z(C1C$!!%1# bO" Z(81$$!!71$
bO"Z(817$!7#1! bO"Z(819$
<È}ä
%
[Y Í-ÇÓ#<È}ä°,
%I%$
)
!
Vé¨ÇÓ#£
!%81# bO" Z(B19$
<È}ä¥
¦¡¬M
!7
~È}ä2
7$1$ bO" Z(B1B$!7$1$
bO"Z("1!$!#"1B bO"Z("1B$
<È}䥦¡·
´
!7
~È}ä2
#917 bO"Z(81%$!#91$ bO"Z(C1C$!
#B1$ bO"Z(C1C$
<È}äc
!
?ÓÑ¿À0
%$
~
È}ä
!
2
#81$ bO" Z(!17$!%"17 bO" Z(!1!$!%81$
bO"Z(!1B$!#B1% bO" Z(!1B$!#"1$ bO" Z(!1B$!#C1$
bO"Z(!1B$!%91" bO" Z(!19$!7#1$ bO" Z(!1"$!!%1"
bO"Z(B1$$!#"1% bO"Z("1B$
<È}ä
!
Ãö
#"1% bO
"Z("1B$
<È}ä{Í-ÇÓ#9Ð
!
,Í-ÇÓ
#<|È}
%
[Y %<ÇÓ#<È}ä°,
%I%%
)
!
VÍ-ÇÓ#£
! 7
~È}ä2
#!1% bO
"Z(B1#$!%C1B bO"Z(91%$!#B1# bO"Z(81$$
<È}ä
¥¦¡¬M
!#
~È}ä2
#$1% bO " Z(!19$!
%81# bO"Z(91$$
<È}䥦¡·´
!B
~È}
ä2
%C18 bO " Z(71% $! #$1% bO " Z(71% $! %"17 bO
"Z(91"$!%918 bO"Z(918$!#"1% bO"Z("1B$
<È}ä
c
!
?ÓÑ¿À0
%!
~È}ä
!
2
7%1$ bO
"Z(71#$!#!1B bO"Z(71B$!7$1$ bO" Z(71C$!%"1" bO
"Z(!1$$!%B17 bO " Z(!1%$!#B1% bO " Z(919 $!#91#
bO"Z(919$!%817 bO" Z(91"$!#$1% bO" Z(91"$!#91B
bO"Z("17$!7%1$ bO" Z("1"$!7%1B bO" Z(C19$!7#19
bO"Z(C18$!#71$ bO"Z(81$$
<È}ä
!
Ãö
#B1% bO
"Z(919$
#91# bO" Z(919$
<È}ä{%<ÇÓ#
9Ð
!
,%<ÇÓ#<|È}
%
[Y 4N8nÓ#<È}ä°,
%I%#
)
!
V%<ÇÓ#£
! 9
~È}ä2
B91$ bO
"Z(B17$!B91$ bO"Z(B1!$!BB1% bO" Z(B1B$!B91# bO
"Z(B1B$!BB1$ bO"Z(91"$!BB1$ bO"Z(918$
<È}ä
¥¦¡¬M
!B
~È}ä2
7$1$ bO " Z(71#$!
#%1$ bO"Z(B1!$!#$1% bO"Z(81!$!#$1% bO" Z(819$!
#$1% bO"Z(81"$
<È}䥦¡·´
!9
~È}
ä2
#$1% bO " Z(919 $! %817 bO " Z(918 $! %C1% bO
"Z("1#$!#%1$ bO " Z("1B$!7%1$ bO " Z("1" $!#$1%
bO" Z(81C $
<È}äc
!
Ñ¿À0
%
~
#71#bO
"Z(81B$
<È}ä
!
`{4N8nÓ#9Ð
!
,4
N8n<|È}
%
76
pV£p
È}ä,Øô¥<+¼N9
!
bÑÒGm
fg0Øô<¥¨j
!
VüÏ<½
&
89<¨&
±8Ï£¤
%
*U£,cÛÅÔ<¶omÎ
M
!
¥¦[Y \Ǩ&±UGö
!
ÛMÈ
}\bLcÛÅÔ©Ð¥
!
P&
!
IMÇ
Ë`°Ì<È}ä°$Â
!
æç\Ǩ&±<
Iª«¬
!
ó04N8nÓ#È}ä°·
Â
!
vcÛUGöæçÆÈ}ä¥<¬
ÁÈ}ä°<·Â
!
ÖÐ|È}<¥
%
5
ËÇË`°Ì:8i½á&
!
¥<,cüÏNO<ÐÜØôiØô
!
V¨&±£
¤<°Ì9¯Øô¬i¥
!
ôIBÓ#È}ä
°$Â
#
bÇË`°ÌÓ#
!
VÇNNФ
<Øôfs¥
!
üÏfs½
#
bé¨ÇÓ#
!
ü
Ïäx½uV\ǵé¨
#
é¨ÇUe¨Ç
cÛ±Í-Ç
&
%<Ç
&
4N8n
!
\Ç<\в
&
Ç
&
Çß
&
%<°ÌíÜ<¨&±¦¡
!
M<
°Ì9¯Øôfs¥
!
ï
7
vÓ#:æç
%$
~
&%!
~
&%
~È}ä<¿À
7
~
~
&9
~È
}äc<À^
!
fg0b\Ǩ&±UG
öÆØôf8%
66
SYZcÀ
#71$ bO"Z(918$
<È}ä
"1% bO " Z("1B $
<È}ä
& #B1% bO " Z(919 $
#91# bO
BB
& ( )
!"
"
6
%6
[Y \ÇcN<
9
vcÛÓ#òöomÎM
2MP1%6&/:=UMX3:4>R/P3@3?M?M@YOXUM/@ =@Y Y3W30/Z:3@U/VW%)3*:#&%88*&=/+*;/%
%5
ÇË`°Ì
# #5
ÇË`°Ì
# 75
é¨Ç
# !5
Í-Ç
# B5
%<Ç
# 95
4N8n
#" F%#5
st{
% F9
<È}ä¶omÎM
%
%5*/@I3:4>R/P3@MXX=0O?"*)$ # #5):4>R/P3@MXX=0O?")$ # 75T0/4O0=>3:4>R/MY# !5+/>Z3Y/3:4>R/MY# B5/UR03Y/@=>R3:4>R/MY#
95f3P3@3>=UMW3Z0=@U# " F%#5#IJ303XU>/ZQ/>3?M?/V% F95
"Z(919$
<È}ä
G# bO" Z(81B$
<È}ä:{
ÇË`°Ì
&
Í-Ç
&
%<Çò4N8n9Ð
!
Ù:ª{#mLÓ#<|È}
% SPM>/M^
"%889$
YZcÀ
B8 bO
È}äb8"Ü\Ç<é
¨Çö
!B9 bO
È}äb8b#±ZÇö
!7# bO
È}ä¿Àb9#±ZÇö
# +=@
"#$$$ $
cÀ
!! bO
<È}ä,@²³\ÇcNan<
#
+XQ/>4=YQM3W3
"#$$$$
b(~
""%:9;/81;*(#&%%$
öcÀ
79 bO
<È}ä,ÇË`°ÌcN<
%
ÙjÍ?<9~Í?
9Ð<
|È}ÏÍ£?
!
ïÏ,9~k<Ñ~¥À
%
SYZXÑ¿
Úfa<
!
{b%"¨m)¦[Y
\Ǩ&±<³Õ}ÒØl
%
+ ,
C=
!
¸è*
!
÷7A
5#$$"5
N9äaS89
(((
[Y
5
öB
&+9N
!"%$ 87 F8!5
Kâú
!
E·
!
ÖÀú
!
5#$$!5
[Y <\üÏÇ,cN
5
89
NÉ)*Ñ
!!$"7$ 7%% F7%75
9B
6!
#
# Kâú
[Y \Ǩ&±UGö<È}ä°
Kâú
!
G-J
!
E·
!
5#$$C5
[Y \üÏÇcNö.<å<
üϽ)Òy
5
&()
!!!"C$ B# FBB5
Kâú
5#$$B5
[Y \üÏÇ,cNò¨&±³Õ
ï&M);))yp[
! B! FB"5
ÙQö
!
iÑ/
!
÷0ß
5#$$"5
Í?T4[Y ~[Ö×°
òS<²½âo
5
&()YZ
!#$"#$ %87 F%8"5
4Ý
!
1ÿ;
!
Þ23
!
5%88"5
@²³
""%4:48:%&*% -5$
Ç
üÏÈ}<qYZ
5
]^N9)k
!7$"%$ %7 F%C5
,@M0.&! G=>:/@ ,! f=/H&5#$$$5&Z=UM/IU3:Z/>=0=XXO:O0=UM/@
=@Y =XUMWMUR /V X=0XMO:IY3Z3@Y3@U Z>/U3M@ bM@=?3? YO>M@P
3:4>R/P3@3?M?!?33Y Y3W30/Z:3@U!=@Y P3>:M@=UM/@ M@ ?=@Y=0\//Y5
<0=@U
4M/XQ3:MX=0=@Y XRU/0/PMX=0:=>b3>?M@ 3:4>R/P3@MXX=0O?XO0UO>3?
/V:=M_35<0=@U30f3Z! C"#$ 9" F"$5
+=@ & H! H=:=Y=G5#$$$5(@MUM=0MY3@UMVMX=UM/@ /V=ZQ/?ZQ/Z>/U3M@ UQ=U
=ZZ3=>?U/43M@W/0W3Y M@ UQ3M@YOXUM/@ /V?/:=UMX3:4>R/P3@3?M?M@
X=>>/U5<0=@U30f3Z/>U?! %8"C$ "78 F"!"5
+XQ/>4=YQM3W3[! oYQ=b/W=[ H5#$$$5,@ =XMYMX3?U3>=?3=?=
4M/XQ3:MX=0:=>b3>V/>?/:=UMX3:4>R/P3@3?M?M@ />XQ=>Y P>=??
?O?Z3@?M/@ XO0UO>3?5<0=@U30f3Z/>U?! #$"%$ #C F775
l:3Y=[! 2OgM*! [=@=43m5%88"5[/03XO0=>=@Y 4M/XQ3:MX=0
XQ=>=XU3>M_=UM/@ /V=Z>/U3=?/:3?O4O@MUV>/:>MX3=@Y X=>>/UX30?5
[/0T3@ T3@3U! #BB"%$ %8 F#"5
SPM>/M^f! <=O0?H5%8895Q=>=XU3>M_=UM/@ /V3:4>R/P3@3?M?I>30=U3Y
Z>/U3M@ M@ =0V=0V=5
EFGH
6
y
6
]
"
"B