免费文献传递   相关文献

Proliferation and Stability of the Regenerated Plantlets from the Embryogenic Callus of Tripterygium wilfordii

雷公藤胚性愈伤组织再生植株的增殖及其稳定性


摘 要:以雷公藤胚性愈伤组织为材料,研究继代培养时间对植株再生和基本培养基、生长素浓度及其与细胞分裂素组合对再生苗增殖的影响。结果表明: 在继代培养过程中,愈伤组织再生率和再生苗数经历了一个由低到高再到低的过程,以8个月到1年之间的胚性愈伤组织的再生能力最强,达100%,每块愈伤组织平均形成的芽数也比较多,雷公藤胚性愈伤组织在继代20个月后已经丧失再分化能力。愈伤组织再分化过程易发生变异,从叶形上看,再生苗出现6种类型的变化。再生苗茎段增殖以1/2MS为基本培养基附加2.0 mg L-1 IBA为好,以1/2MS+2.0 mg L-1 IBA+0.5 g L-1 活性炭的培养基有利于芽苗生根培养,生根率达100%。移栽至珍珠岩与腐殖质土(1∶1)的混合基质中成活率达96%。

Abstract: The embryogenic callus of Tripterygium wilfordii was used to study the effect of its subculture duration on the plant regeneration, and the effect of basic medium with different auxin concentrations and their combination with cytokinin on the proliferation of its regenerated plantlets. The results showed that during the process of subculture of the embryogenic callus, the regeneration rate and the number of the regenerated plantlets increased in the first stage and then declined in the second stage. The embryogenic callus with its subculture age between 8 month and 1 year old was most regenerative, with 100% regeneration rate and more plantlet number per callus. The callus had lost its ability to differentiate completely after 20 months of subculture age. The differentiated plantlets from calli were vulnerable to mutate. The plantlets could be divided into 6 types, according to the shapes of leafs of the plantlets. When the plantlets were propagated with stem sections, 1/2MS supplemented with 2.0 mg L-1 IBA should be optimal. And 1/2MS+2.0 mg L-1 IBA+0.5 g L-1 active carbon medium was favorable for the plantlets to root, with rooting rate 100%. When transplanted in the mixture of perlite and humus (1∶1) soil, its survival rate was as high as 96%.


全 文 :! !"" ! ##
$ % % &$ # %
& ’ ( )
’()*+,)- ’)./-* ’)+)(-*
/012!"*+02#
3456*
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
$ % % &
d%6ob1sOnUDSQ0jLmUbpb"
/ *#5$ +,-# >M.$ /0s$ ’ 1#
7#2 N&8&(:;)O%:81PQ+Y8s ~N@DT81ST:BPQ8s ct 7#$#%%>
$2 N&8&(:;)DE())G ct 7#$#%%H
! "# !d%6ob1sOn’@D*PQfJX-F3aSQUDj+WX-+cDx?OgaUD%jL0HI\WS~M=lfJX-ST8*1sOnUDZjUD%087y{nK0I.
UI00ST*! 8n%I #$T30ob1sOn0UDfF/G*! #%%9*’%1sOnR]0R0u0)
[\f*d%6ob1sOnlfJ $%n%|N8y{U6+fF\1sOnU6+ST;cDQk*7Z09
g*UD%zj :|_~0Q+\UD%mxjL! #;$<’’+WX-+9t $2% =>l.? # )@-’)*! #;$<’ A $2%
=>l.? # )@- A %2" >l.? #9bv0X-+O@,u%DVX-*DVZ! #%%9\wW‘*wOQKLJM7# B #H0
]g+J8R9Z! &:9$%&= d%6>ob1sOn>OnX->,$
’()*+#’7$$2CA 7BD&!C2# ,-./0#- ,12+##%%# ? 7!883$%%&4%# ? %%"7 ? %"
>?@#=$%%8 ? %$ ? #8+ABC=K>e}_hOiBC7$%%$@-"#:-%!H" Zl’LMNO!"#$%&’"()%#* (*+ ,)(-%$%). #& )/’ 0’1’*’"()’+ !$(*)$’)2 &"#3 )/’
43-".#1’*%5 6($$72 #& !"#$%&"’(#)* +#,-."/##
.E F45#*$ GH5> 3I5J40# (KH5 LE5MI$ <40 FI45KIE$ NK45> LE5>#
7#2 !"#$%&’&() *%+ ,%(-%"".-%( /"%0". &1 2-&3"40-#-+" &1 5$**%6- 7.&8-%#" 9"4"*.#$ *%+ :"8"’&3;"%0 /"%0". &1 2-&.*0-&<*’ 7"40-#-+"*
=&.0$>"40 ?@A B%-8".4-0) C*%(’-%( 7#$#%%>
$2 /&’’"(" &1 D-1" 5#-"%#"*=&.0$>"40 ?@A B%-8".4-0) C*%(’-%( 7#$#%%H
8-2)"(5)= ,KH H=OPM0>H5EQ Q411IR 0S !.-30".)(-<; >-’1&.+-- T4R IRHU J0 RJIUM JKH HSSHQJ 0S EJR RIOQI1JIPH UIP4JE05 05 JKH V145J
PH>H5HP4JE05*45U JKH HSSHQJ 0S O4REQ =HUEI= TEJK UESSHPH5J 4IWE5 Q05QH5JP4JE05R 45U JKHEP Q0=OE54JE05 TEJK QMJ0XE5E5 05 JKH
VP01ESHP4JE05 0S EJR PH>H5HP4JHU V145J1HJR 6 ,KH PHRI1JR RK0THU JK4J UIPE5> JKH VP0QHRR 0S RIOQI1JIPH 0S JKH H=OPM0>H5EQ Q411IR*JKH
PH>H5HP4JE05 P4JH 45U JKH 5I=OHP 0S JKH PH>H5HP4JHU V145J1HJR E5QPH4RHU E5 JKH SEPRJ RJ4>H 45U JKH5 UHQ1E5HU E5 JKH RHQ05U RJ4>H 6
,KH H=OPM0>H5EQ Q411IR TEJK EJR RIOQI1JIPH 4>H OHJTHH5 8 =05JK 45U # MH4P 01U T4R =0RJ PH>H5HP4JEYH*TEJK #%%9 PH>H5HP4JE05
P4JH 45U =0PH V145J1HJ 5I=OHP VHP Q411IR 6 ,KH Q411IR K4U 10RJ EJR 4OE1EJM J0 UESSHPH5JE4JH Q0=V1HJH1M 4SJHP $% =05JKR 0S RIOQI1JIPH
4>H 6 ,KH UESSHPH5JE4JHU V145J1HJR SP0= Q411E THPH YI15HP4O1H J0 =IJ4JH 6 ,KH V145J1HJR Q0I1U OH UEYEUHU E5J0 : JMVHR*4QQ0PUE5> J0
JKH RK4VHR 0S 1H4SR 0S JKH V145J1HJR 6 ZKH5 JKH V145J1HJR THPH VP0V4>4JHU TEJK RJH= RHQJE05R*#;$<’ RIVV1H=H5JHU TEJK $2% =>l
.? # )@- RK0I1U OH 0VJE=41 6 -5U #;$<’ A $2% =>l.? # )@- A %2" >l.? # 4QJEYH Q4PO05 =HUEI= T4R S4Y0P4O1H S0P JKH V145J1HJR J0
P00J*TEJK P00JE5> P4JH #%%9 6 ZKH5 JP45RV145JHU E5 JKH =EWJIPH 0S VHP1EJH 45U KI=IR7# B#HR0E1*EJR RIPYEY41 P4JH T4R 4R KE>K 4R
&:9 6
9’. :#"+2= !.-30".)(-<; >-’1&.+-->H=OPM0>H5EQ Q411IR>JERRIH QI1JIPH>P4VEU? VP0V4>4JE05
d % 67 !.-30".)(-<; >-’1&.+--H’ d e (
7(H14RJP4QH4HHd%6]ST*’_"xySTT{*
7’TE5>1H "0 *’ 6*#&!#>.I0 "0 *’ 6*$%%!>^zd*
$%%8H\(F*Mc[{|G"01vST*!O{
12Gc9|LAc’}y~cxys!c0}"c0
1G0Nv72h#d*$%%7>/KK,d%6’f$D1J6W*DjO%:8’DQ90;(4v*rRqDDEN
G’v\STiZX-jSQUDZ[0^_[+
8STE|0G"SN7&"3d*$%%$H\LS1
sOnUDr?zjOb[Qk[STOnX-S
T8B~kl0jS7^_Ad*$%%8H\aSTZ
fYOb[0fYcUDSQmU|J0PQ~M*
l1mQk8*O’!Qk[aSTb/~ROF
0*-V+!bp]2*8x*lSTE|9OK[
04v7’**d*!""#>5^od*!""$H\a]*C
*ad%6dde(ST0PQ_"/8lH|$
LcOX,$7Tzd*!""%>"(Rd*!""%>2-
Rd*!""#><>d*!""#Hdop*i,d%6On
X-UDSQ0PQK5.;*8xO""ad%
6OnX-UDZ[0^_*+[3PQ\WPQ
!d%6ob1sOn’@D*PQfJX-F3
aSQUDj+WX-+cD6x?OgaUD%jL0HI*!#’d%6L
S1sOnr?XEOb[QkAt|lwxQ%
&opyuvw& @DQo4
!"! tzCm
7<^@)zy/q/D&$DRo**.,ct;s%L9*A.^! &" ’(F*;ZmTmvvkzv+-v‘
r7ZYzv|*{7yv ! )*%"*,z’! !" +*
Os7yv #r*& ,l-. & /,01! ’! # (23*Os7
yv #r\4:B-u0mxvNMSZ*+-u
{VPQ\ZhR "4$ ’( 5 "4$ ’( o%q|l
67 8 &4" (,l-. & !*#9: 8 "4$ (,l-. & ;!$ = >" ?|0R1sOn\1sOnl(gX-+
9|vX- >J*0R0ob1sOn}v\ob
1sOnl 67 8 &4" (,l-. & !*#9: 8 "4$ (,l-. & ;<
X-+9*fJ’k*>" ?fJ &r!"# tz{|
&4!4& b89:bE;<=>;3?g12 j
v 67c&@!67 ;(o?_qHc&@#67 ;(o?’
670 &@#HcA$c/cBC6 D|+WX-+*]kt &4"
(,l-. & EAF\’n{wq| #".*’.q| &nB
-u0mx\X- >" ?|*AB/uMSZ0c-
un0c=3<^*F-/uZ*/uZ G/uMS
Z0 5 &""@7q|MSZ0 ./[MSZ0H&4!4! @kAB\CDEFGHI3JKLMNO ! 67’+WX-+kt "4& (,l-. & HFF 8
&4" (,l-. & D9AF*PQX-F3ad%6ob1s
OnSQUD0HI\qX-1sOnsql’(
STDn6+QFy{PQ\’n{wq| >".*’.q
| >%ob1sOn\X- >" ?|*ABcu1s
On%0c’%1sOnR]0Run0*-F-6
+Z*6+Z G6+1sOn%0 5 &""@q|1s
On%0&4!4> @kAnUD0R0 $|0%jaqKu{V0R00%
’@D*l 67c&@!67c&@#67cA$c/cBC6 D |X-
+9*kt !4" (,l-. & EAF*PQ+WX-+ad%
6u%fJjL0HI\l &@!67X-+9kt’
(U^0 EAFcHFF*PQ’(DujD<0HI>l9t !4" (,l-. & EAF0 &@!67
X-+9kt’(U^0 D9AFj ;<*PQDQfY6x?Ogad%6u%fJjL0HI’n{wq| >".*’.q| &nMSZ*X- $"
?|*AB/uZcu%.^cDVZdXY&4!4# S\TUV{EsOn0R00%*fJX- >J!|*l9t !4"
(,l-. & EAF0 &@#67X-+9*kt’(U^09
bv*PQ9bvau%DV0HI\’n{wq
| >".*’.q| &nMSZ*X- $" ?|*ABD
V0cVD<_‘*F-DVZbvX-+*t;CD >" ,l-. &*v0n % ,l
-. &|+*I/A‘ $4J*l; 7^!$ K !HL*|E’.
&! )*|EG^’ & """ = & $"" 1M0X-sX-F ! Tz{|T(}S~!"#>U)wB-.!
$%&’! ())*+,- .) &%-/+ 0*1/% .2 %3/44%56 &71 /217+,/.2
%21 85.9,: .) ! ’ "#$%&’(##
+WX-+
AN+2’ (O?2P(
/uZ
QOR(23NS2T3
RNS2T@*
=3<^
E3SOR3T?O
1O3,S)@’(
u%.^
/O2,)S TU
I1N3S1OS@’(
67 V%4## W "4J# ’ #4"J ’
A$ V$4&! W’ &4#% N J4>" N
/ &""4"" N "4V% ’ D4J# W
&@! 67 &""4"" N &4!" W %4&$ W
&@# 67 V!4$" ’ "4>D ? >4&% ?
BC6 V&4D% ’ "4!% ? !4V# ?
!~80w’ >rGY0R]-\’($N~i8 :P3’N3
bAY($v$$k+{7! X "4"$H\z(\:NSN 23 S)O ’T1P(3 NRO
S)O (ON3+ TU S)ROO ROIOS2S2T3+Y :2UUORO3S 1OSSOR+ 23 S)O ’T1P(3+ +)TZ
+2,32U2’N3S ?2UUORO3’O NS ! X "4"$ W[ :P3’N3P+ (P1S2I1O SO+S Y <)O +N(O
WO1TZY
! WSQ67
#"! Tz{|T(}S~!"$JU)wB-.
K~ &+U*||X-+8-u0/cZ]l
V"*!9*-u%0D<_‘$k\;\8dU^
OK|0 &@!67c/X-+’%O@,-u0/c
ju0D<*/uZ! &""**Vu%0=3\<*
ng*{GfJX-\.U^OK|0 67X-+
90R0u%*ZYY[*D<0=*-u%=3C*’@,*{GfJX-\.1jk0 A$ X-+
90R0u%*ZYY[*DJ$ & ’ ( ) #$"
)’@,*{GfJX-\]0OK|U^0!"#
$%c&’$ X-+8*-u/c|D):([)*2+)v67B-.
d%6ob1sOnl $%9t ()) *+! ,-l
./ ! 0 123) !+* ,-l./ !X-+9*fJ|! 4.n[
2;*! 5* .|*1sOnCbQ[*zjuH*!
n%|u%0R\7! 6Jn[Ou6+*VWf
Jr00jt*6+ZCbT.\7 8 !5 n%T3
6+Z! !**9*’%1sOn6+z0u%0!
!7nHI*V6+z00%ZYY[*D+Zj’%1sOn90R0u0)CbzF7~
5H*1sOn2;M+*6+z00%30ZT.fJX-! !7n%01sOn2;|#Cbs[*
%n11sOn94*5*n%|N/0y{y6+fFd%6ob1sOnlfJX- 5*n%|*s
ql $%9t *+*4 ,-l./ ! ())>*+! ,-l./ ! ())>
*+4 ,-l./ ! ())>*+4 ,-l./ ! 123)>!+* ,-l./ !
123)>5+* ,-l./ ! 123)>*+*4 ,-l./ ! ()) 0 *+4 ,-l
./ ! 123)>*+! ,-l./ ! ()) 0 !+* ,-l./ ! 123)>*+5
,-l./ ! ()) 0 5+* ,-l./ ! 123)>*+4 ,-l./ ! ()) 0
4+* ,-l./ ! 123)0X-+9**+’(STD=.Ogopz*d%6ob1sOn6+QFy
{PQ\t#LS~?AUob1sOn6+fF
056*||{w0X-+9]}Ou6+\+5*
d%6ob1sOnfJ 5* n%|Ny{6+
fFF ! {|’u()*2+;<,)B-.
#$%&! ’(()*+ ,( *-.+-/) +01) ,2 *$..-3 4.$2+.)+ /)5)2)/$+0,2
,( ! & "#$%&’(##
X-F3
:;<=;>?
=@,?"
,AB=C
q|1s
On%0
(;,D?> AE
FG<<@
@BAF;QDo/Z
01sO
n%0
(;,D?> AE
FG<<;I
?,D>JA@HI
@BH;F?H
6+Z
KG=? AE
H@EE?>?B=@G=@AB"9
’%1sO
nR]0R
un0
$?GB
B;,D?>
AE D;HI
L?> FG<<;I
5 7M #! #1+*M F M+!M H
# 74 NN M*+1# D !5+#4 F
7 M# M# !**+** G !N+MN D
!5 7N 7N !**+** G !7+45 G
!1 MN #7 #M+#N F 6+!# ?
!7 M* !# !4+41 H !+5N E
5* M* * *+** ? *+** -
!"6 }S~([)*2+,);<%b:@@A[
lkty *+! ,-l./ ! ()) 0 !+* ,-l./ ! 123)
0 $%X-+9*l 7n%I !$T3*d%61s
On6+Zqr !**9*0%D00%7Z09g*O!z 1|_~=! !|7#(
! / !HZ0’<\0*ZkO-78>! 5|7#(
! / 5HZ0’9o0*ZkOf78>! 6|7#(
! / 6HZ0’oij0*Zk0k>! #|7#(!
/ #HZ0’<\0*ZkOf78>! 4|7#(!
/ 4HZ0’9o0*Zk0k>! 1|’aqKu
{V0R0SQ7#(! / 1H*Z0’o\0*Zk
Of78\8SfJfJX-|*1sOnUDS
QZ0jZkQo6+z00%,[b/bp*6
+z00%7#(! / !HD<0=*m/-x*ZY
Y[!"7 }S~"‘Bj-
5+#+! b89:bE@kA||X-+9u%0/cZ]\.7~ 6H*-u%
0Du%D<\)*u%=3<^)\ngfJX-$%j 34 X-+90R0u%D<0=**U 34 X
-+9u%D=3\C*u%D#$%&6 ’(()*+3 ,( %$30* 1)80$ ,2 $90..$/: %-8
3-%*-.+-/025 1-.+04.0*$+0,2
+WX-+
3GI@F
,?H@G
/uZ
P?>,@BG=@AB
>G=@A"9
u%.^
O?@-C= AE
L=3<^
QB=?>BAH?
jL60
KG=? AE
,;<=@L<@FG=@AB
DVZ
KAA=@B-
>G=?"9
$% MN+## D 4+16 F *+N6 F 4+5! F !6+66 ?
34 M1+MN D 7+41 D !+5M G #+!# ? !5+** E
O !**+** G 7+6! D *+N4 F 1+*7 D 57+5! H
!"5 $% !**+** G 7+MN G !+!# D N+15 G #4+** F
!"# $% M5+4* F 5+76 H *+4N H #+M7 H 17+#5 G
&’$ M!+1N F 5+## ? *+#6 ? 4+!5 FH 15+4* D
5+#+5 Q3); ())E;-+8kt’(U^0 Q3)|*’%au%0/c
mD#H\l *+4 8 5+* ,-l./ !T3*/uZ.! !**9*D
VZmV0D<_‘)VWU^0T.]jtQ3)U^’ 5+* ,-l./ !F=3\<*ng*{Gf
JX-\Q3)U^’ *+4 ,-l./ !FO[u0R-=
3\C*’ng,fJX-\? Q3)U^’ #+* ,-l
./ !F*au%0D6+jD<\’t~?0X-+9*u%r2}<*
ZB"?*Cbz\M4! !# ^ zd=d%6ob1sOnUDSQ0jLmUbpb
F ! "#$.!(!"#>U)wB-.
%&’(! )**+,-. /* "#$ /0 &1233&45 ’67 2076,-2/0 &07 84/9-:
!"#$
7%&l’( )H
/uZ
*+,%-./0-1.
,/0-1$2
u%.^
3+-&40 15
67/.07+0$8%
=3<^
!.0+,.19+
7+.&04$8%
jL60
:/0+ 15
%;70-67-8/0-1.
DVZ
:110-.&
,/0+$2
<=< >?=@< 8 )=@) + <=AB 9 >=<< 9 <=<< +
<=@ )<<=<< / C=?D 9 <=@D 8 @=)=< )<<=<< / D=@< E <=BA E D=FB E A@=F@ E
C=< )<<=<< / B=?> / )=)C / ?=DF / >C=<< /
>=< F>=BF E >=FA 8 <=?F E D=D@ E CD=F? 9
G##U^l <=< H C=< %&l’( )T3*u%0/c
Zc.^c=3<^cjL[0cDVZ*VW G##U
^T.]jt7~ @H*-D<_‘]’mt;,(
U^0 !"#\? G##U^’ >=< %&l’( )Fu%0D
U)wB-.
%&’(; )**+,-. /* <$$ /0 &1233&45 ’67 2076,-2/0 &07 84/9-:
G##$
7%&l’( )H
/uZ
*+,%-./0-1.
,/0-1$2
u%.^
3+-&40 15
67/.07+0$8%
=3<^
!.0+,.19+
7+.&04$8%
jL60
:/0+ 15
%;70-67-8/0-1.
DVZ
:110-.&
,/0+$2
<=< >?=@< 9 )=@) + <=AB 9 >=<< 9 <=<< +
<=@ F>=B? 8 C=>> 9 <=@> 8 @=)=< )<<=<< / >=>F E <=B< E D=>> E CA=<< E
C=< )<<=<< / D=?@ / <=B? / ?=?C / C>=<< /
>=< F?=CC E C=BA 8 <=>C 9 @=AB 8 )>=F? 8
C=>=A !"#[ DI"#; JK\]^_E12 K~ D+5*l !"#U^’ C=< %&l’( )F*t
;’(U^0 DI"#j JK]’@,u%0D<*~
jl/cZc.^c=3<^cjL[0cDVZ]0
,aE*VWU^0jt*/uZcDVZdCbz
F = "#$a =>#$U ?%/G0m(!"#>U)wB-.
%&’(= )**+,-. /* 20-+4&,-2/0 /* "#$ &07 =>#$E?% /0 &1233&45 ’67 2076,-2/0 &07 84/9-:
~?U^ 31,%1.+
81.8+.0,/0-1.$7%&l’( )H
!"# DI"# JK
/uZ
*+,%-./0-1. ,/0-1$2
u%.^
3+-&40 15 67/.07+0$8%
=3<^
!.0+,.19+ 7+.&04$8%
jL60
:/0+ 15 %;70-67-8/0-1.
DVZ
:110-.& ,/0+$2
C=< <=< <=< )<<=<< / B=?@ / )=)> / ?=?C / >C=<< /
C=< <=@ <=< )<<=<< / >=@A E <=?F E D=D) E )D=D? E
C=< )=< <=< FA=AA E A=>F 8 <=DB 8 @=AA + )@=<< 8
C=< C=< <=< BD=D? 8 C=)F + <=@> 9 >=DB & )A=AA 9
C=< <=< <=@ B<=<< 9 A=C) 9 <=@F 9 D=)B 8 ))=?D +
C=< <=< )=< DD=D? + C=)C + <=>@ + @=@) 9 <=<< 5
C=< <=< C=< D)=CF 5 )=AA 5 <=C< 5 @=)> 5 <=<< 5
F\JK0$VNv\ DI"#G*t; <=@ %&l’( )JK*
/uZcDVZZ61zFy C<2j A<2@A; u[1jk9("‘)2B-.
lDVX-+8*t;{pU^09bv’%
y.yDVZ*V0D<_‘)M+),aE7~
?H\?9bv’ <=@ &l’( )F*DVZ! )<<2*V0
D&l’( )au%DV/’O@\U^@S <=@ &l’( )*
DVZc’QR]V0c%0.^cD<_‘*VW9
bvU^0jt]F0\+f[8’9bvO\G
0K9fF*K9yX-+80;(8-TJb{F B u[1jk9()2B-.!
%&’(B )**+,-. /* &,-2C+ ,&4’/0 /0 4//- 2076,-2/0
9bvU^
#80-L+ 8/,E1.
81.8+.0,/0-1.$
7&l’( )H
DVZ
:110-.&
,/0+$2
R]V0
M+/. .;%E+,
15 ,110N
u%.^
3+-&40 15
67/.07+0$8%
V0D<_‘
*,1O04 N0/0;N
15 ,110N
<=< DA=AA 8 )B=C< 9 ?=>) / P P P
<=C@ )<<=<< / A)=D< E D=?< E P P P P
<=@ )<<=<< / >B=>< / D=@< 8 P P P P P
)=< ??=>C E CF=?A 8 @=FA 9 P P P P
C=< @)=@C 9 ))=?@ + C=?@ + P P P
! P P=D<{m *+.+,/7 &,1O04>P P P=D<) *119 &,1O04>
P P P P=D<\) "+00+, &,1O04>P P P P P=D&,1O04Q
@A= 3‘:45
lDVt2%. D H B 8%Fn[:%*qLq
.x,n&;sL;|z|X:% C 9*:z0%*
v‘r7vzV‘00n*Uv B<<6 C@2fs<
Q=V‘ @ %-.*a|wW‘8.;}s0*wOj
KLJM7) R )H08->8*v )$)< MS %?2*@
95D>’G\’=#? ) rfs%*)< 9T|L’5D>*D< C@ 9HI+BMw
WI.3*:%wWR9Z!I F@2A Ix
WPQ8d%6ob1sOnUDSQfFl
fJ8zjLCbj.*UF0*fJ Cy{6+fF*fJX-8UD0SQZ~zjf
|M+0Q+*1Z&y1sOnfJX-0’b
pb\ZfYoUDfF0y{[STOnX-8
{n_"0mn*lJfSTOnX-8VW1s
On8f6ZjFP6Z0(0jtV{UUDf
FzF0‘y{7Z3Ad*C<yu7 !"##$%&’( )&*#’+’(H72!Td*)FF>HcqB
7,&+*’#H7Z3Ad*C<8)O_*0.;\VW1sOnfJr00j
@"
t*f0+8~01sOnUDSQfFzF*!0
+8~01sOnUDSQfFxOjtw{a’
ul4{7R7!"# !" #$ $*%&&’H\$c5d7(’’)H
PQ~M*lbCX7%#&’()’# (’*!*(’(Hob1sOn
fJX-86+fFzFQfJX-80R;(f
6ZmFP6ZfYOi>^?-d7(’’*HQ’[
YZ0CQk[ZfYOb[Qk0_"‘8\O
id%6ob1sOn<#fJX-8fYQk0
‘8qOx,*{GPQ*a1sOnX-8ZfYOb[QkV{S
QUDfF0zF*-1mQk;f0[+]20
7|Cgd*(’’)>^ _Ad*(’’+H*a,ST]2~
Rr;*+N’Qk0{|ArE7,"-."/0-.0*
(’’%H\LS-u{V0LpcDr?lT%b/
9[bp0*]r?’uy‘@D0]2bpbd%6ob1sOn0UDSQZ0zjy||Q
+*-V8SmxjLX-|*UDSQZ0jZk
0Q++bpy’(zr\UDSQzj01|M
0Qk*ULlOfR6[GcDQ+*LS*{G
0PQ+’ij.%(9bR60k[qp+R9bvad%6u%DV!OM+0P*N
v\WPQlDVX-+8t;9bv*’%+!
P*d%6u%’pVQD*)+!P*V0D<*
-V+$VVDQjS0cD\1lEUFd
7(’’1Hdl-#:7+’*,( #-.#*/’’H’pV0DVX
-8).zy_*0WS\-GInd7(’’)HlK
F701&21-# 3#&1*’)#H0DVX-8!cj9bva
DVO{p0$VNv\DVX-8*9bvlX
-+8!Oyuv\|{c0X-+8 233 j 243 0Fs75#667/ !"
#$ $*%&&’H\9bv+!K9X-@DQD0A_
TJ*syDQjS0cD*y.yMSZ0R9Z
QLpcDfF\9bve+!K>jsK9ST
~?cD_jA]gE*1|Fj[bK9aX-@
D+fgQD]Nv*OnX-8Jp9bv0K
9QOPTJO+,ws9bvlSTOnX-8
aSTD$c5*Sjo*dY"*d $(’’) $bCXob1sOnfJX-S
T80fY[YZ0CQk $N&ST).*()7+H=%1’’ 8
%1’9 $
^?-*4HQ $ (’’* $QRob1sOnmUZocDST8[Y
Z0C0Q+ $<^8&;)).=‘a()(**(79H=9:+ 8
91*$
EUF*^ z*Z I $ (’’1 $-#:RJo0LpcDPQFH $
&’&’;)).*(+79H=%’’ 8 %’:$
^ z*K3L*2gY*d $ (’’+ $d%61sOn{Vmxy9b
PQ $N&8&(:;)).=‘a()(**17:H=%’* 8 %’+ $
^_A*[ M*,_M*d $(’’+ $STZfYOb[QkPQ*s $
ST)L.*(:7%H=%(% 8 %(+$
"(R*NpO*e_4*d $ (’’) $PM#VQ0OnX-Q,m$
L $STDw)LM*9*7%H=%(+$
|Cg*R S*^ nT $ (’’) $STZfYOb[Qk:B0PQ*
s $KN8’()**+7*H=(*+ 8 (9*$
< >*PUV*2UA*d $(’’9 $W‘6t2%DVX-PQ $ST
)L.*(%7*H=**( 8 **1 $
5^o*a P*^ bF $ (’’: $STZfYOb[Qk0PQQ4
v $W8).*%&71H=9)& 8 9+9$
2h#*J^-*;U‘*d $(’’) $d%6LXBZLM0}"Nv $
<^$(;)).*9%79H=*’9 8 *’)$
2!T*^ %9*Z;F*d $ %&&9 $|yuob1sOn0Qkm.
2-R*SdB*X7q*d $ (’’9 $Yq6UDZ[0^_ $&Z)
.**%7(H=(9% 8 (99$
’***S2K*2_B $ (’’9 $STZfYOb[QklSTb/~
R804v $STDw)LM*9’71H=)1* 8 ))%$
T z*eU&*[rG*d $ (’’) $de]STOnX-Os%0o
4PQ $n&&SPQ*((7(H=%9’ 8 %9($
GIn*Z\n*< K $ (’’) $KFSQUD:BPQ $K#8’(
)**:7*:H=%%9%+ 8 %%9%&$
&"3*JCA $ (’’( $1WSX]2s+PQ*s $SX).*%&
7*H=%&% 8 %&+$
Z3A*e[=*]IA $ (’’1 $qB1sOn6bQ+mUQZfY
owcDfF0i[ $]2).***7)H=19) 8 1:9$
/K<*Z5(*C^G*d $ (’’) $d%6+gTa1o_2‘ab
|2DR0HI $8N$Wg).*:7:H=:%& 8 :(’$
!"# ;*4<=>7? @$ %&&’A B>"/= ?7C7/7?"=#0/ D?0E 7EF?G0C7/#H H">><6 #/#=#"=7I
D?0E #EE"=0?76H7/H76 0D 67J7?"> .#C.K="//#/ 60?C."/=
!7>>*;#66<7 "/I L?C"/ !<>=@<0 M H0EQ0D?0E %-’&"!-45’,. 6’$71-/’’ "H=#J7 "C"#/6= 84"2’.*# (!&#-#"! $ O
5"=5#667/ , R*,<==7? S T$%&&’ $ ,="F#>#=G 0D 233 "/I 243 #/ /<=?#7/= E7I#67J7?"> =#66<7 H<>=,"-."/0-.0 U V*O#QD 3*W"X"67-"?"/ Y*!" #$ $ (’’% $ 2/IG
7EF?G0C7/#H H">># "/I Q>"/= ?7C7/7?"=#0/ #/ ZQ>"/I791((4&’,. 2’-(,",.
@$H"/I B#E"791((4&’,. :#-:#/!*(! @$H!0==0/6$ !?0Q ,H#7/H7*9%
7*H=%(*: 8 %(9’$
,[#/C>7 \ ;*U">>7? U @*,#7C>7? S U*!" #$ $ %&9% $ !.#/767 #/67H=#H#I"> Q>"/=
%-’&"!-45’,. 6’$71-/’’ #/=?0I