SCAR Marker and Detection Technique of <em>Bursaphelenchus xylophilus</em>-文献传递-植物通论文库

将松材线虫RAPD特异片段OPM₀₅-X₂₁₀₀进行分离、回收,与载体pGEM^®-T Vector连接,转化大肠杆菌并培养,对目标克隆测序。根据测序结果,用Oligo5.0软件设计引物,正向引物为M₀₅F₂(5‘-CGGGT CATGG CTGGA GGTAT CGT-3‘),反向引物为M₀₅R₁(5‘-TGGCT CAATG GCAAA TCCTT CGTA-3‘),成功地将松材线虫特异片段OPM₀₅-X₂₁₀₀通过引物对M₀₅F₂/R₁转化为SCAR-M₀₅-X₆₀₀。运用SCAR标记引物M₀₅F₂/R对枯死松树体内分离的92份线虫样本的DNA进行标记,并对单条线虫经简易方法提取的DNA进行检测。结果表明:引物组合对所有81份松材线虫株系均扩增出一条600 bp的清晰、明亮的条带,而对8份拟松材线虫、1份霍夫曼尼伞滑刃线虫、1份大核滑刃线虫、1份长尾属线虫均无扩增产物。该对引物可以检测单条松材线虫。利用这对特异引物组合可构建松材线虫检测试剂盒,实现对松材线虫的快速检测的目标。

OPM₀₅-M₂₁₀₀, the specific RAPD fragment of B. xylophilus , was collected from agarose gels and purified. Then, the purified fragment was inserted into the pGEM^®-T Vector that was transformed into E. coli and cloned and sequenced. Based on the sequence of RAPD marker, the sequences characterized amplified region (SCAR) primers were designed by the aid of the software Oligo5.0. The forward primer is M₀₅F₂(5‘-CGGGT CATGG CTGGA GGTAT CGT-3‘),and the backward primer is M₀₅R₁(5‘-TGGCT CAATG GCAAA TCCTT CGTA-3‘). The specific fragment (OPM₀₅-M₂₁₀₀) was successfully converted to SCAR marker (SCAR-M₀₅-X₆₀₀) by using M₀₅F₂/R₁,which was the specific markers of B. xylophilus. Then, the DNA of 92 isolates of Bursaphelenchus, B. mucronatus, B. hofmanni, Aphelenchoides macronucleatus and Seinura sp. which were isolated from dead pines, were marked, and the DNA of a single nematode extracted with a simple method was detected using this set of specific primers. The results indicated that the PCR product of all 81 isolates of B. xylophilus was a clear and bright fragment about 600 bp with M₀₅F₂/R₁. But eight isolates of B. mucronatus, one B. hofmanni, one A. macronucleatus and one Seinura sp. had no any fragments. Assay M₀₅F₂/R₁ also successfully detected single pinewood nematode. Therefore, the specific pairwises would be used for constructing identification kits of B. xylophilus, implementing the aim of quick detection, and achieving the purpose of identify juvenile successfully.

全文：!!"
" !
#
#
$ % & $
$
#
%
& ( )
()*+,)- )./-* )+)(-*
/012!"!+02#
3456!$ % & $
lØP²
(-T
¡!
j
7
L
7
U
7
F
7

7
D^
"
ØO&R)E&STJHI)D
7
½VgÌFöµçJë ¯
7
ØO
$&%%#>$
!
7
"
!7
,lØP²
T-`A
ÈªNS
e`3
%;
@K
$&%%
4®=
%
§£
!
JCC
Pa*3
)
@,/Láô
!
(=R,-^
«³<
!
<«¡Q
&
;<mn
!
j
e1IG0;2%
¦§}Dö
!
ÙDös
3
%;
B
$
";]Y(aaa,(-,aa(,aa-
aa,-,(a,Y#]$!
×ÙDös
3
%;
T
&
";]Y,aa(,(--,aa(---,((,,(a,-Y#]$!
:àw,lØP²ÈªNS
e`3
%;
@K
$&%%
¹ZDö<
3
%;
B
$
ZT
&
(=s
(-T@3
%;
@K
9%%
&
Gj
(-T
¡3
%;
B
$
ZT
<#lÉC_=
e
8$
WP²2Be
A+-
4®¡<
!
«<^P²Ûdïs4e
A+-
4®
&
mnop

Dö9´"&
WlØP²-$y~Ü÷^
9%% QP
e/º
%
pÚe^^
!
<
"
W%&
W~G»:P²
%&
W
RÝ:P²
%&
WÿkuP²yW~ö
&
é&
µjø<ÈªDö9´±ò~
lØP²ïúû
!
&
#$%
7
lØP²
# (-T
¡<
#
ÈªDö
#
±ì
&()*
! >9#2&&777
+,-./
!-777
+01*
!&%%& @>!"""$%&$#%# @%%"" @%>
£¤¥#
$%&% @%> @$!#
¦§¥#
$%&% @%8 @$!&
¨©ª«

ACZ[()¨©ª«
"#&&>%;88$#
½V2DZ[()BC¯Rª«
"&%mk-$$%%$&$#
ØO&R)D42x´i¦(B
¨©ª«
"a$%%8 @%8$&
!Ü~±s¹ºµ»
&
!+:2M&]*&/08*(*)(#"/C*)3/#\6*"9@"$&89#,#:%9"&0/,+89),"&
(JLF BLFG=407_LkI4F5LF7SD KI40[IF7CD4FG.IF7,4FGkI4cIF
"8.2#3#./0.&#$+"#$.-&#$%*7 1*5,&.*6#*+! P%*;,*3 0.&#$+&4=*,5#&$,+47e,%*3$- J#4K%C.&%+.&4/.&
!#*+,.* %*7 L%*%3#6#*+./)*5%$,5#$
:<.(&)(7e`3
%;
@3
$&%%
! MJLNPL,JLF! MJLPD5IOILH O54G=LFMV4NIFNL5MLH IFM0MJLPa*3
)
Y,/LNL[DLFHLNIGFLH QWMJL4IH 0OMJLN0OMV45Le1IG0;6%6,JLO05V45H P5I=L5IN3
%;
B
$
";]Y(aaa,(-,aa(,aa-aa,-,(a,Y#]$!
4FH MJLQ4%;
T
&
";]Y,aa(,(--,aa(---,((,,(a,-Y#]$6,JLNPL%;
@3
$&%%
$
V4NND<%;
@K
9%%
$ QWDNIFG3
%;
B
$
ZT
&
!VJIA42.B(,2-$@,JLF! MJLA+-0O8$ IN014MLN0O:-&$%B(#2#*(-$!:@6-&.*%+-$!:@(./6%**,!9B(#2#*(.,7#$6%&.*-2#%+-$
4FH <#,*-&% NP6VJINI=P1L=LMJ0H V4NHLML:@A42.B(,2-$V4N4<1L454FH Q5IGJMO54G=LFM4Q0DM9%% QP VIMJ 3
%;
B
$
ZT
&
6RDMLIGJMIN014MLN0O:@6-&.*%+-$! 0FL:@
(./6%**,! 0FL9@6%&.*-2#%+-$4FH 0FL<#,*-&% NP6J4H F04FWO54G=LFMN6-NN4W3
%;
B
$
ZT
&
41N0ND<NIFG1LPIFLV00H FL=4M0HL6,JL5LO05L! MJLNPLA42.B(,2-$! I=P1L=LFMIFGMJL4I=0O[DI=*> 4"0.7 :-&$%B(#2#*(-$A42.B(,2-$# (-T=45dL5# NPL77(-T"NL[DLF¡<Æ
4`54F
Å
"&88#$
9îe÷Êvü=x¡<
&
¹Z<¸#e
T-`A
NSeQJ
!
«<
×4®=W}Ü÷<ÀRÎ
T-`A
NSUV
e
$!
d~¨eDö
!
jLDö<ÎÄe;
A+-
4®
(`T
~
!
È"×~\æ
"(-T$
ìÈª
~
&
z
(-T
¡VEFÚÊ
%
qÊeÅ]
%
ÊvAjÊx+/
ïÀg¿veyj
" 4`54F #+%2@! &88##
-H4=YR10FH0F #+%26! &88!# a45eJ=05I#+%26! &889# C4W=LN#+%26! $%%%# m4N4I#+%26!
$%%%# /IH41#+%26! $%%!#
¶¤¥Å
! &888#
[¥Å
!
7!
#
# ?ÇÅ

lØP²
(-T
¡$%%%#
kÅ
!$%%&#
¶²gÅ
!$%%##
f^Å
!
$%%#$&
|P²AïÀ
!3LFG
Å
"$%%!$
:àw,
Øï;mP²
"L#2.,7.34*#,*.*3*,+%$
j;m
P²
"L@;%5%*,%$
Èªe}h~1G
A+-
N
S(=s
(-T
¡<
&
j
(-T@` (T
Dö<±q
/qwAØï;mP²
%
;mP²juF;
mP²
"L@%&#*%&,%$!
«T~/q/q
&Z#
^
$
oã²
%
d²õ²
&
B,jlØP²Èªe
(-T
NS
!
}Èª
Dö
!
<#lÉC_=eR?P²
A+-
2B4®
¡<
!
«A+-
4®
!
ïüw¶Üg»elØP²=x±ìJïs
&
&7
ØOJïs
A@AB
Q|O
P²tÏÞèAÁÉ
%
¿S
%
Ø
%
¯È
%
z~
%
½
%
¿Ç
%
yN
%
aÅà1dVWe#lÉ`
=V
!
²=P²ÄZ¥B
%
A
%
5R
%
ÎA
q;@Aew&
ïP²µ
8$
d-$
!
G@l
ØP²
":@A42.B(,2-$$"&
d-$
!
":@
6-&.*%+-$$ "
d-$
!
~G»:P²
":@
(./6%**,$ &
d-$
!
RÝ:P²
"9B(#2#*(.,7#$
6%&.*-2#%+-$$ &
d-$
!
ÿkuP²
" <#,*-&%
NP6$&
d-$
"
o
&$&
A@GB
&nb5
R,-^
k3&%8
^-Jv8CC
Pa*3
)
@,
/LZ
5`0=LG4
¬
&
A@?B
þ"Än
ïúJàq
,4[ A+-
_´ÀZRáoFö
¬
# aL1*XM54Z`,ØFöj
¬g3¬
# )`,a"
ª½¨ø×Â3
$! K@a41
"; @

@! @
B
@# @

@
5
@I@
×Â3
$%
¼
ÌRº
"-=PIZ
5`0=LG4
¬
&
³<¨
.R".D5I4YRL5M4FI$
³<¨
"& %%% =.$!
>¼ÌRºe
.R

!
>
-=PIa41
e
.R

! e(
³<¨
&
A@HB85:
Þß
R?
A+-
4
& $
Ï·e|P²çC
$%%
+
.
z
*PPLFH05O
K@
!
F
$%%
+
.
P²}Dò
!
g
# $$
|
9; n
+@5
& J!
é
&% =IF
n
&
´
# #$
§8Zn
!
jÅCÁ
"!%%
+
.$
e"
-4
!! n
^§ç
!&$ %%% 5)=IF
@&
)
$% =IF!
`
/ò
# !$
jÅCÁ
"!%%
+
.$
e"
q
B·
q
ªó
"$;q$!q&$
-4
!
)
# ;$
jÅCÁ
"!%%
+
.$
e
B·
q
ªó
"$!q&$
-4
!
)
# 9$
|`/ò@
F
&Z&%
CÁe
# =01).
@&
ef
" PCj!29$
j
$
þCÁeW+fó
" @$% n$!
|
@"% n
^§ç
é
$% =IF# >$ &$ %%% 5)=IF
@&
)
$% =IF!
åêj
>%^
efó
" @$% n
µ
$
¾
$
´nk
!
åê
`k
& J# "$
§fóHî£
!
F>g
T+-
Àe
&%%
+
.,*
¯våê
! T+-
Àeÿ/s
$%
+
G)=.
@&
!
Ï·z
@$% n
j
& 8$

;
+
.
¥&
$%%
þ
!
j)â=ââ/
A+-
eÿ/
&
?P²;^P²
A+-
4
&
V^P
²
!
F
&%
+
.
µP²}DòeY
!
,P²J}
Dò×z
*PPLFHH05O
K@
&
|
(`T
`
!
z
>% n
^
§çKn5
#; =IF#
|
8; n
^§çKn5
&% =IF#&$ %%% 5)=IF
@&
)
$ =IF!
`/òjz~
&
A@IB
<
2:%8
Ê
&2;2& 7 T-`A
ÕÖ
7
}hDö
e`3
%;
;]@
aaa--(a,a,@#]&
&2;2$7T-`A
×±ØB
7
×ye´òfC
Ás
$;
+
.!
G@
&% b` (T
êìò
"%2; =01).
@&
e
m(1# &%% ==01).
@&
,5IN@C(1! C`j82%# &^ ,5IM0F
K@&%% $ $2;
+
.! $2; ==01).
@&
e
H+,` $
+
.!
$; ==01).
@&
e
3G(1
$
$
+
.!& o,4[
À
!$ ==01).
@&
eDö
&
+
.! &% FG)=.
@&
e;
A+-&
+
.!
20
^¯{+
&92;
+
.&
~s
,`(@$%% A+-*FGIFL&
&2;2#7T-`A
×±Ù
7
×y´ò|
8! n
µ

; =IF#
4FJH
8! n

& =IF!#9 n
*
& =IF!>$ n
[÷
$ =IF!
µ
!%
dJH
#
³
>$ n
[
÷
&% =IF&
&2;2!7T-`A
ÚÛg
7
×ymþ
!

"
+
.

~ö|
&2;^
!ÃÂ"¸@
&%% /
º0^§çº
#
&"% =IF!
j=fv
&% :&; =IF!
|
R)e@
T-A
"¸:q$¥`î.
%
V
&
A@XB
<
2:%8
Ê8Da

%28^
e!ÃÂ"¸
!
J¸£±èûE
&
,
T-`A@` (T
ö|!ÃÂ"¸`º#Z«eN
SJGaNS=9
!
|)çjW^eMN
,«eNSÜçÄ
&
j
aL1*XM54<
«eNS4®§£
&
A@cB
<
2:%8
Êns ¡
$
îa
&2>2&7
ËÜ
T-`A
ÝÞßBàá
7
,
,@
CC
jA+-"<0FM501IFNL5MA+-$
µt$)
&
áô!Ñll
$ b
áôêìò
&
&$
|^e
%2; =.
e)K@=Fç×
:=
$ b
áôêìò
;
+
.!,@
CC
";% FG)
+
.
@&
$
&
+
.! (`T
§£ö
$
+
.!,! A+-
áôÀ
"# o)
8"
& ( )
!"
"
7
+
.
@&
$&
+
.!
éx+ZoCÁ
&%
+
.&
$$
j?´òlly×yò
!
náô
& J!
Ë±|
! n
çáôZ
"! n
çáôZ±
F³Re(=»F
$&
>
AB
|BC
C&
L[DLFwÒ
(0HL
Êø
PLÄT
e5IGIF

L[DLFwÒ
(0HL
Êø
PLÄT
e5IGIF
& (&! @; RX
¥B
k4P4F !> aE& RX
¿S
aD4FGH0FG
$

$ a0FG[I$ RX
¥B
k4P4F !" aE$ RX
¿S
aD4FGH0FG
#
wÔ
# ,I4F<40# RX
¥B
k4P4F !8 aK# RX
¿Ç
aD4FGXI
!
Ý
# ,4FGcIF # RX
¥B
k4P4F ;% k.& RX
½
kI4FGND
; o-& RX
A
o- ;& k.$ RX
½
kI4FGND
9 (-+& RX
5R
(4F4H4 ;$ k+a& RX
½
kI4FGND
> --& RX
ÁÉ
-FJDI ;# k+a$ RX
½
kI4FGND
" --$ RX
ÁÉ
-FJDI ;! k+a# RX
½
kI4FGND
8 -3-! RX
ÁÉ
-FJDI ;; k+k& RX
½
kI4FGND
&% -C$ RX
ÁÉ
-FJDI ;9 k+.$ RX
½
kI4FGND
&& -C# RX
ÁÉ
-FJDI ;> k+E& RX
½
kI4FGND
&$ -CB& RX
ÁÉ
-FJDI ;" k+E$ RX
½
kI4FGND
&# -3& RX
ÁÉ
-FJDI ;8 kEA# RX
½
kI4FGND
&! -3# RX
ÁÉ
-FJDI 9% kEA! RX
½
kI4FGND
&; -+$ RX
ÁÉ
-FJDI 9& kEk& RX
½
kI4FGND
&9 -.& RX
ÁÉ
-FJDI 9$ kEk$ RX
½
kI4FGND
&> -.$ RX
ÁÉ
-FJDI 9# kEk# RX
½
kI4FGND
&" -,# RX
ÁÉ
-FJDI 9! kEk! RX
½
kI4FGND
&8 -,! RX
ÁÉ
-FJDI 9; kEk; RX
½
kI4FGND
$% -S$ RX
ÁÉ
-FJDI 99 kEkT& RX
½
kI4FGND
$& -S# RX
ÁÉ
-FJDI 9> kEkT$ RX
½
kI4FGND
$$ ka! RX
Ç
kI4FGXI 9" kEkT# RX
½
kI4FGND
$# C_& RX
yØ
CDF4F 98 kEkT! RX
½
kI4FGND
$! C_$ RX
yØ
CDF4F >% kET& RX
½
kI4FGND
$; C_# RX
yØ
CDF4F >& kET# RX
½
kI4FGND
$9 C*& RX
yN
CDQLI >$ k(& RX
½
kI4FGND
$> C*$ RX
yN
CDQLI ># k($ RX
½
kI4FGND
$" _S$ RX
Ø
_DFF4F >! kk& RX
½
kI4FGND
$8 _S# RX
Ø
_DFF4F >; kk$ RX
½
kI4FGND
#% a# RX
P0
I9 k+& RX
½
kI4FGND
#& a! RX
P0
I> k+$ RX
½
kI4FGND
#$ (E& RX
¯È
(J0FG[IFG >" kK& RX
½
kI4FGND
## ($ RX
3S
J4FH0FG >8 kK$ RX
½
kI4FGND
#! EB! RX
a
EJLcI4FG "% k(& RX
½
kI4FGND
#; EB; RX
a
EJLcI4FG "& k_& RX
½
kI4FGND
#9 EC& RX
a
EJLcI4FG "$ k-` %& R=
¥B
k4P4F
#> EC$ RX
a
EJLcI4FG "# meT%& R=
ÎA
0DMJ m05L4
#" E+& RX
a
EJLcI4FG "! ,S%& R=
e
,4IV4F
#8 E+$ RX
a
EJLcI4FG "; CR%& R=
yN
CDQLI
!% EE& RX
a
EJLcI4FG "9 k%# R=
½
kI4FGND
!& EE# RX
a
EJLcI4FG "> Ek%& R=
a
EJLcI4FG
!$ EE! RX
a
EJLcI4FG "" -C%# R=
ÁÉ
-FJDI
!# aA$ RX
¿S
aD4FGH0FG "8 C+%& R=
yØ
CDF4F
!! aB& RX
¿S
aD4FGH0FG 8% B$- RJ
P0
I!; aB$ RX
¿S
aD4FGH0FG 8& 3e -=
A
o-
!9 aT$ RX
¿S
aD4FGH0FG 8$ k(&&- N
½
kI4FGND
77
!
RX
lØP²
:@A42.B(,2-$# R=
:@6-&.*%+-$# RJ
~G»:P²
:@(./6%**,# -=
RÝ:P²
9@6%&.*-2#%+-$#
N
ÿkuP²
<@NP6
&2>2$7
âáãÖäd
7&$
ÜÏ·z
@"% n
e
k3&%8
¸bG Ò×zÊ`óZ2=
"
ð
; =IF$!
l¡q´Ï·ò
!
)(´
9%
+
.k3&%8
¸bG
Ò
&2; =.
W^
*`
K@
!

$
+
.
áô×yò
!
%8
7!
#
# ?ÇÅ

lØP²
(-T
¡×zÊ`
# $$
|Ê`l¡j
!
Ê
$% =IF# #$!$
n
«
!; :;% N"
G&*èn2
$# !$
8¤Ê
$
=IF# ;$

8;%
+
.
n
e(
³<¨
"
Ë±j
.R
³
<¨
$!
g
!
|
#> n
n¢@
&;% 5)=IF
@&
£³<
&2; J& 9 $
,×z
#> n #% =IF
e
-=PI)`,aZK@a41.R
F
&%%
+
.
£³ò
!
j¤¼yg¤¼
!
Ó×z
#> n
¥nê@
³"&9 :$! J$!
ÿÜü
Z
¦
!
>gøFNS
e(=^ès¦
!
5>ZáCCe(=^
ü¦
&
A@WB
b5
85:
8Þß
3ä
4=Q500d
Å
"$%%;$
ïs
&
A@SB
¢KÊ8£e
jx]lÅ
"$%%$$
ïs
&
j~«eNSe
ÈªDö<(= Ò
A+-
4®~
&
º#~
NSR
!
«J«eNS
%
(= Ò
A+-
4®
&
§~NSÿ/J(=eÈª
T-`A
NSÿ/8
!
T(= Ò
A+-
e³´FIz~NS
A+-
q;
(=Èª
T-`A
NS
!
Ý«eQ
&
A@ARB
dé
,(=|e Ò
A+-
¨`,Ã{Fö±ìg
3¬
!
j
-R)
$¥
!
¹jDö
,>" 5`0=0ML5
,--,-(a-(,(-(,-,-aaa$
j
9` " 5`0=0ML5
(-,-(a-,,,-aa,a-(-(,-,-a$
4®
&
A@AAB!+:2
çè8nª7
;<
T-`A
ÈªNSQemn
!
j
e1IG0;2%
¦§}
(-T
Dö
&
}|eDö`,Ã{F
ö±ìg3¬´:
&
A@AGB!+:2
-¤8
%+2

×yC$
$2;
+
.&% b (`T
êìò
! &
+
.
$2; ==01).
@&
e
H+,`! &2;
+
.$; ==01).
@&
e
3G(1
$
!
& o,4[
À
! &
+
.$ ==01).
@&
eÊDö
! &
+
.$% FG)
=.
@&
e;
A+-!
j20^¯{+RCÁZ
$;
+
.&
×y¬

×y´ò|
8! n
µ
; =IF#
4FJH
8! n

!; N!!8 n
*
#% N!>$ n
[÷
9% N!
µ
!%
dJH
#
³
>$ n
[÷
> =IF&
×yö·z
! n!
| ¸Z¬@
!
F
;^
e
ae.A/)*S
O
"
i=f
$!&2#^
e!ÃÂ
"¸º#
!&%% /
º0^§çº#
9% =IF!
|
R)e@
T-A
"¸:q$¥`î.
%
V
&
$7
mnJ=W
G@AB
<
2:%8
Ê
Dö
e`3
%;
e~äü
&&
|lØP²|-$
¶&
"
#7
& :8$
y~Ü
&
^ÿð
$ &%% QP
ýþ
epÚ/º^^
!
"
#7
&% :
&9$
¶&yWL^^
&
¹L
!
ls}hDö
e`3
%;
slØP²eÈª}hDö
!
~ÜlØP²eÈ
ª
T-`A
^e`3
%;
@K
$&%%
&
ü
&7
j}hDö
e`3
%;
~e
$ &%% QP
Èª
T-`AA+-
NS
"
vÑ_
$
BIG6&7$ &%% QP NPLO54G=LFMNDNIFGP5I=L5e`3
%;
"NJ0VF IF MJLVJIMLMLXMQ0X$
3 &%% QP A+-14HHL5# & :8
lØP²
:@A42.B(,2-$"(&! @;!
o-&!(-+&!--&!C_&!C*&!EB!!aA$! k.&$#&% :&9
ØP²
:@6-&.*%+-$"CR%&!Ek%&!k-`%&!,S%&!meT%&!
C+%&! -C%#$6
G@GB
<
2:%8
Ê(¥
$
Danâã
,
T-`A
ÈªNS
e`3
%;
@K
$&%%
4®=
%
§
£
!
JCC
Pa*3
)
@,/Láô
!
(=R,-^
!
F>¼ÌRº
%)`,a%K@a41
e³<¨@³<
&
4®ü¦3E
"
ü
$$&
|ü¦3Ee¨N`
!
4÷4j
(`T
s¯9xv8
&
Hü
#
M?

¯
9xv8
(`T"
#7
#$
RJ
e`3
%;
@K
$&%%
"
#7
$$
R÷ú
& Pa*3T@,/LRs
# %%% QP!
GJÈªNS
e`3
%;
@K
$&%%
¶j
"
#7
!$
¤¯9x
v8
A+-!
³´FÏIz#7
$
j#7
#!
op«e
NS¢øFCC
!
Ý«eQ
&
ü
$7
Èª
T-`A
NS
e`3
%;
@K
$&%%
(=mn
BIG6$7,54FNO05=4FMN0ONPL%;
@K
$&%%
O54G=LFMN
¦>gøFNS
!
ç¦5>ZáCCe(=^
SJIML<010FILN<0FM4IF IFNL5MN0ONPL%;
@K
$&%%
O54G=LFM!Q14G@?B
U|<Êéê
,NSe¯9xv8
A+-
¨`,Ã{Fö±
ìg3¬
&
ü
#
s
e`3
%;
@K
$&%%
e×
&
é
NSµRs
$ &%% QP!
NSÿs
$ %8$ QP!
|×
;]
VïÑ_çC²=s}hDö
e`3
%;
e
&8
& ( )
!"
"
7
77777
ü
#7
ÈªNS
e`3
%;
@K
$&%%
¯9xv8
(`T
ü
BIG6#7TL<0=QIF4MI0F P14N=IH (`T34PN0O
NPL%;
@K
$&%%
O54G=LFM
&
¡¢
A+-A+-14HHL5!O50=(06 5`0=LG4#$
ÈªNS
e`3
%;
@K
$&%%
PL%;
@K
$&%%
O54G=LFM# #
¯9xv8
(`T
TL<0=QIF4MI0F P14N=IH A+- (`T# !
¯ 9 x v 8
A+-
TL<0=QIF4MI0F P14N=IH A+-6
~¨×
!
|
#]
VïÑ_]C²=s}hDö
e`3
%;
~¨×e×ÙÀR×
&
|
e`3
%;
e~¨
×J×ÙÀR×¶&ÿ/s
$ %&9 QP&
G@HB!+:2
-¤8lm
;<
e`3
%;
@K
$&%%
×
!
}
&
^ÙDöj
&
^×ÙDö
!
9:Dö<
&
ÙDös
3
%;
B
$
;]@
(aaa,(-,aa(,aa-aa,-,(a,@#]!
Rz
e`3
%;
@K
$&%%
×e
;!" :;>% QP

!
äü
!
çC²=
#
×
ÙDös
3
%;
T
&
;]@,aa(,(--,aa(---,((,,
(a,-@#]!
Rz
e`3
%;
@K
$&%%
×e
& &$! :& &!> QP

!
äü
!
]C²=
&
z×ÙDö|}hDö
e`3
%;
×eÀR×_Ö
!
¹LDö<
3
%;
B
$
ZT
e
~NSÿ/z
e`3
%;
@K
$&%%
×eÿ/
&
Dö
<
3
%;
B
$
ZT
&
eµ#~NSÿ/s
9%% QP&
Dö<
3
%;
B
$
ZT
&
e*n/s
!8 n&
~mnäü
;&
ü
!7e`3
%;
@K
$&%%
×
BIG6!7L[DLF%;
@K
$&%%
$8
7!
#
# ?ÇÅ

lØP²
(-T
¡ü
;7
Dö<
3
%;
B
$
ZT
&
~P²e
(-T
¡<
BIG6;7(-T=45dL54=P1IOILH VIMJ P5I=L5NLMN
3
%;
B
$
ZT
&
O50=FL=4M0HLN
&
¡¢
A+-A+-14HHL5#$ :&%
lØP²
:@A42.B(,2-$
"(&! @;!o-&!(-+&!--&!C_&!C*&!EB!!aA$! k.&$#
&& :&"
:@6-&.*%+-$"CR%&!Ek%&!k-`%&!
,S%&!meT%&!C+%&!k%#! -C%#$6
77
ü
;
±ä
!
lØP²2B
"
#7
$ :&%$
Uè
~Ü
&
^ÿs
9%% QP
e^^
!
T|2Be^^
U8./º
%
pÚ
!
"
#7
&& :
&"$
yWL^
&
¹L
!e`3
%;
@K
$&%%
¹ZDö<
3
%;
B
$
ZT
&
±(=s
(-T@K
9%%
!
slØP²ÈªNS
!
(-T@3
%;
@K
9%%
&
G@IB
?@N?F|gz
ÞR?P²@4
A+-!
ÛDö<
3
%;
B
$
ZT
&
~
!
GüHü
9
M?
&
Dö<
3
%;
B
$
ZT
&
||l
ØP²-$&
"
#7
& :"&$
yg÷pÚ
%
/ºe
9%% QP
^^
!
"
d-$
"
#7
"$ :"8$
yWÛ-^^
!
~G»:P²
"
#7
8%$%
R
Ý:P²
"
#7
8&$%
ÿkuP²
"
#7
8$$
ËA
Ü
9%% QP
^^
&
¹L
!
Dö9´
-
|R?2q
oÜI2eÈª
& $
w,lØP²J%
~G»:P
²
%
RÝ:P²
%
ÿkuP²\9Ä
&
¹L
!
±
q=j
$
&
ü
97
jDö<
3
%;
B
$
ZT
&

8$
W2qmn
BIG697(-T=45dL54=P1IOILH VIMJ P5I=L5NLMN3
%;
B
$
ZT
&
O50=8$ N4=P1LN
3
¡¢
A+-A+-14HHL5#& :"&
lØP²
:@A42.B(,2-$#"$ :"8
:@6-&.*%+-$#
8%
~G»:P²
:@(./6%**,#8&
RÝ:P²
9@6%&.*-2#%+-$#8$
ÿkuP²
<@NP6
G@XB
u¦|àgz
2q
& :9
ÆÛZdïs4e^P²
A+-!
jDö9´
3
%;
B
$
ZT
&
~
!
Gmn=äü
>&
Dö9´+2|
#
dlØP²-$&
"
#7
& :
#$
yg÷pÚ
%
/ºe^^
!
#
d-
$
"
#7
! :9$
yWÛ-^^
&
opDö9´
3
%;
B
$
ZT
&
èÜ^lØP²
&
P²*+ÛZo
pe¬4
A+-!
Ë*y|©4®Ü
!
±
óôj}Dò}D4®~
!
ýµdz
!
%´F
`yj
&
#7
m6J76
lØP²
T-`A
ÈªNS
e`3
%;
@K
$&%%
¹ZD
ö9´
3
%;
B
$
ZT
&
:à(=s
(-T
¡<
"(-T@
3
%;
@K
9%%
$&
ÙDös
3
%;
B
$
";]@(aaa,(-,aa
(,aa-aa,-,(a,@#]$!
×ÙDös
3
%;
T
&
";]@
,aa(,(--,aa(---,((,,(a,-@#]$&
ü
>7
jDö<
3
%;
B
$
ZT
&
^P²ü
BIG6>7(-T=45dL54=P1IOILH VIMJ P5I=L5NLMN
3
%;
B
$
ZT
&
O50=NIFG1LFL=4M0HL
3
¡¢
A+-A+-14HHL5#
& :#
lØP²
:@A42.B(,2-$"(&! @;!o-&! --&$#
! :9
:@6-&.*%+-$"CR%&!Ek%&! k-`%&$6
¹Z<#lÉC_=e
8$
WP²2q4
®
(-T@` (T

!
lØP²emnJ»G
mnKk÷ú
&
Dö9´|lØP²-$&yo
g÷^
9%% QP
e/ºpÚ^^
!
Dö<²
%
~G»:P²
%
RÝ:P²
%
ÿkuP
#8
& ( )
!"
"
7
²e~öyWé^^F
&
z
(-T@` (T
mnÆ>Ð¡!
¹L
!
DöªI/
!
mnzz>Ð
&
jé¡<ïs&±q|t&_®A
&
4
A+-
e&ð
;% =IF# (`T
~ð
& J"
HjHE~
(`T

34NML5
~&ó4÷4t
$#
º#&ð
#% =IF&
èdZ¬*¯Z
$2; J!
ÝE«¡
&
j
(-T@` (T
£K±q<^P²4®¢
A
!
;|F`AAyÏª/«#lÉÆi
¹lØP²¬Ìú
!
õÆlp¨´ö_Æi>
glØP²Åûå
&
#lp_õlp¨´ö_
AAg÷ÊP²õÆOÊP²
!
¹L
!
+<#l
É_eP²õÆlp¨´ö_eP²4®kÀA

&
Hn55!
g±
èùúG
&
f=P²?I
!
~Z,?P
²k²}D
&
f=P²?1
!
~Z}h-
P²}D
&
Þ
T-`A
Èª^^3E
%(-T
(=Bï
eP²
!
MEj2BgÄZA
%
¥B
%
5R
q;èAtÏ]\elØP²
!
&gÄZ¥B
%
ÎA
q;èA²=w\e!
ÄZzèAtÏ
l&=¼\_#lÉ`e
#
ÊGaP²
!
2BB
g÷eo
&
ÞïmnÄð
!
lØP²e
¢Fq
&%%^!
mn¢
%
±³
&
£KHL
!
µ
s÷Êïúû
!
&yÏ¹Z{1eP²2B4
®Õ
&
Z [ + ,
k
7

!
§{
!
A
7

!
Å
6$%%&6
phÆ®
+
¯p
9

,
Èª×
~¡<
" (-T$
~8;G|pÊA@eyj
6
µö)6
!
$>"9$ >$$ @>$"6
x]l
!
§°&
!
h
7
±
6$%%$6
=xFö)56J±ì
6
NO

NO
()±ìÜ°·
!$%6
¶²g
!
®AË
!
Ðâc
!
Å
6$%%#6
÷dJX²]^WN¨¹
9H"@
!,G
=
áÈe
(-T
¡6
ëö]5)6
! ## " $ $
&;& @&;;6
¶¤¥
!
Gò
!
3_`
!
Å
6&8886
År]¨¹
!6VU
e=x
Aj¡6
û0)6
! $9 "9$ 9># @9"$6
[
7
¥
!
®,À
!
(A³
!
Å
6$%%%6
Ç´Å#¿]ÚÊ=x¡EFeBC
6
û0)6
!$> "$$ &;& @&;>6
f^
!

7
Ã
!
Ðµ¸
!
Å
6$%%#6
ÅYÕø;P²¨¹
"G* @
6*2
e
(-T
¡<
6
yjJHIFö)6
!8"#$ $;% @$;#6
4=Q500d k!TDNNL1AS6$%%;6
=xQØ
6#
°
6
¯³¶
!
Ð
·¸
!
8hNÅb
6
NO

()Ü°·
!$> @$86
-H4=YR10FH0F -B!LUIGF4<3! R4FFL50MC!#+%26&88!6(-T! T-`A
4FH TB.` =45dL5N1IFdLH M04H0=IF4FMGLFL"9&#$ <0FOL5IFG
5LNINM4F"""9Z>$ "9; @">%6
a45T-`A! (-T! 4FH TB.` =45dL5NMIGJM1W1IFdLH M0FL=4M0HL
5LNINM4F(4B.3%#%6aLF0=L! #8";$ "#9 @"!;6
C4W=LNm3! /4F HLSLGS*! -5LFN` ! #+%26$%%%6ALUL10P=LFM0O
(-T=45dL5N1IFdLH M04!(4+.B(+(.&% /&%3%&,%#5LNINM4F4FH MJLI54NNLNN=LFMIF *D50PL4F 4FH +05MJ -=L5I<4F NM54VQL5W
GLF0MWPLN6k0D5F410OMJL -=L5I<4F 0m4N4Im! 305Id4V4_! 05I/-! #+%2@$%%%6ALUL10P=LFM0O(-T
=45dL5NM0MJL` /_5LNINM4F%73
Q4NLH 0F 4<0==0F OL4MD5L
0OP14FMHINL4NL5LNINM4F3LFGl !` .0FGC! KD kC6$%%!6 (`T4NN4WNO0554PIH 4FH NLFNIMIUL
IHLFMIOI<4MI0F 0OMJ5LL=4c05500M@dF0MFL=4M0HLN!L#2.,7.34*#
,*.3*,+%! L@;%5%*,% 4FH L@%&#*%&,%6-IFI<4! #!"#$ $%! @$&%6
eJ=05I,! 3D54M43!30M0W0NJIB6&8896301LT-`A4FH (-T=45dL5N1IFdLH M0MJLS6@& 10,JL05-PP1aLFLM! 8$"$$ &;& @&;96
4`54F )! 3I=45dL5N1IFdLH M0H0VFW=I1HLV5ININMLF-PP1aLFLM! ";""$ 8"; @88#6
/IH41kT!AL14U4D1M` !(045L53!#+%26$%%%6ALNIGF 0OG54PLUIFL"H,+,$
5,*,/#&% .6$ P5IFMIFG6,JL05-PP1aLFLM!&%&""$ &&8! @&$%&6
!
!"#$
7
åæç
"
!8

SCAR Marker and Detection Technique of Bursaphelenchus xylophilus

松材线虫SCAR标记与检测技术

相关文献