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.�/*� ���� 01���� 234)Brassica napus L.( ��7� �8&�1& 0���� 9���� +��� .�$�� ��12 �����
=���� �� +�$ .������ ������� �����) ��Cauliflower mosaic virus, CaMV ������� ����� � (
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� ���� T��� ������ �� �!" TuMV �CaMV ��& ����� � U��'� �� �!" ����� ���� ���� ���� =
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+X�� ������ : =���R� =0���@�Brassica napus
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:��8!� EF'%��� �� )Brassica napus
L.( �. �� '% ,!%�>%�. �:��0G�� :�� 8!� EF' H!%
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�$ ��%�� OP0JK� Q81� �� L�� �. � O>+ �
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1985) ��� �8�� �. )�� �$ .12 �. �$( 9�8!�
�� %��� �$ �� �. $�!%�' R5+ )�� �% �� �6�.
&�� S�!%�0� 9�8!� T�Cauliflower mosaic
virus (CaMV) � &U�� S�!%�0� 9�8!�Turnip
mosaic virus (TuMV) � 9�8!� �.8V �$��
�6( ��6UWBeet western yellows virus
(BWYV) �$J7(% 8X5 �% �87��. :� % �%
�5%$�0Y8. )Coutts and Jones, 2000;
Farzadfar and Pourrahim, 2014;
Shahraeen, 2012; Tabarestani et al., 2010 .(
9�8!� &�� S�!%�0�T� )CaMV( Z05[ H7�%$ .
�$)%�% P�$ �%�5% . $��L 8000 �. :\��50� ]�;
/6�Caulimovirus �� ��.)Hull and
Shepherd, 1977 &!^� .(CaMV _�% �$� 8�;
`�0. )Brassicaceae� ( � S�!%�0� ���8�� T�
�� �<a0;0���. . b0�-� ��$0a+ �$ &!^� H!% ��
72
:,-. / � � $%� &'(9�8!� �!%�� S! �. %��� Z(�% ,6�%�...
TuMV ) :'% 8;$0R��Walsh and Tomlinson,
1985; Melcher, 1989 .( $��L27 �7� �50��. -
B�� � �5!" CaMV &a' �� �. $0a+ �� S! �% %�
�� Ic75%�6$ )Shepherd, 1981)+ )�� �$ .( �$
�50�Myzus persicae (Sulzer) �
Brevicoryne brassicae Linnaeus &R� H!8;
T(5� �7Y6� �� 9�8!� H!%���6�. )Walsh and
Tomlinson, 1985 8d�% .(�!%�� � R6; 9�8!� H!%
_�% ��$0a+ �. �$(� `� 8�;0. ���6�. $%�1; �
�1 �% �$���!%��� �50� �$ )+ /6� e�7-� �
�� ��$0a+ $�!% )0;0;) �66�Schoelz and
Shepherd, 1988 .(�. ��$0a+ CaMV � ��� f5�
0 5�� ���5%$ �$�. � $0�. �6V�� �� ; ��+ :'$
50 �g�$�� ,�) �.!Sutic et al., 1999 .(
B�%��� �.!$� �% �$�17�CaMV �% e�7-� hG6�
�� :'$ �$ )%8!%9�8!� H!% i0�� 8<5�5 �� ��. �$
�0�� ) :'%Sohi et al., 2013 .(
%� �% ��!��O�*Y ,� � CaMV I876�
�7�I' H!% �G �% :'% )+ T(5 � �$ O%8�L
T.c�c� Z0 '� $%$ )�5 :�) �5%Edwards et al.,
2008�!%�� Z ; 8<!$ j8G �% .( . 9�8!� H!% �
�7�� 5 Ic75% H!% �. Z�c� �%��� Z(�% H7>! .�6.!
)[ ! 9�8!�_�% �$ :��c� �� `� 8�;0. %�
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`� 8�; )�� ,6�%� �$�17� Oc�cK; �G 0.
8.%8. �$CaMV �'�8. ����� � �5-�� m!%8� �$
�� �� )�5 Oc�cK; H!% n!75 .:'% ,6�%� �� �$
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�� O�\7� 8<!��!_�% �% �Y8. �$ R6; � ��.� H!%
�. T K7� Z(�% $%05YCaMV :'% �� :>! (Pink et al., 1986; Pink and Walkey,
1988; Provvidenti, 1980; Saunders et
al., 1990; Walkey and Neely, 1980;
Walsh and Tomlinson, 1985(.
�. :��c� /�*"���.%�+ Z(�% �% �Y8. �$ H�6o
�!%�� �Y8. �� ��� 9�8!� H!% � H!% � :'%
�$ :��c��!%�� �. ,6�%� 8<!$ �CaMV �7*��
:'% �� )Agama et al., 2002 pCallaway et
al., 1996 pCecchini et al., 1998 pHapiak et
al., 2008 pTang and Leisner, 1997(.
��6!% �. ��0; . )06� ; �$ %��� ���; Z(�% ,6�%�
)%8!% �.CaMV �7>8<5 �%8( �.!��% $�0��$ �:'% h�cK;
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B�%�� �$ ��6!% �. ��0; . H�6o� �0_L �$�17�
)�� CaMV �TuMV :'% �� ��!; �� �$
)Farzadfar and Pourrahim, 2014 p
Shahraeen, 2012 pTabarestani et al., 2010 .(
)�� ��$0a+ 8.%8. �$ %��� ���; Z(�% ,6�%�
CaMV �TuMV �� �'�8. ��5.
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�5* T�� h�cK; H!% �$ �� $\7'% 9�8!� ft6�
:50\�%�� �!%�� CM1841 9�8!�CaMV Z6.
pCa122 )Anderson et al., 19919� . (� �
V00140 � �!%��TuMV .9�� �Ku535893
$0.. .�:50\� �5* �% $\7'% �0X6�%�� CaMV %�7.%
5* � pCa122 �.�!0' Escherichia coli DH5α
Tc76����87�. �% .8; �%Z8\*5�� '^" �� u%8-7'%
�� )Sambrook et al., 1989(. �� '^" /�'
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73
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Table1- Name and characteristics of canola varieties evaluated in this study Name of Varieties Farming area Type
Source
Hayola 401 Warm-humid zones Spring Australia
Zafar Warm-humid zones Spring Iran
Eureka (Sary Gol) Warm-humid zones Spring Iran Karaj1 Cool and temperate zones Winter Iran
Karaj2 Cool and temperate zones Winter Iran
Karaj3 Cool and temperate zones Winter Iran
RGS003 Warm-humid zones Spring Germany
Opera Cool and temperate zones Winter Sweden
Talayeh Cool and temperate zones Winter Germany
Licord Cool and temperate zones Winter Germany
Okapi Cool and temperate zones Winter France
SLM046 Cool and temperate zones Winter Germany
Zarfam Cool and temperate zones Winter Iran
�!��5� �� ��� ���. �n!��; �$ S� 7*�' &!^�
v8. :>! B87*� �!P. �)Lebeurier et al.,
1980 ��!; �%8. .(v8. �$ �� I1> 9�8!� �
�$ �&U��)%v8. �% T� �%�!P. � �!��5� ��5
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p6- forward )5´-
GAGAACATAGAAAAACTCCTCAT-
3´ ( �CaMV-p6-reverse )5´-
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bakhsh et al., 2007(.
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u8� Z(�% � :�%$3��!^G �%8"�% � � �"��%
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�,$1- ��� �� �$�& ��NO��� ������� ����� � ������ �&� ) ��Cauliflower mosaic virus, CaMV .(A ���� :
= ��NOB = �*�^ :C � ��$� _�W� :D _�W� � ��`���� :.��$�
Figure. 1- Symptoms induced by inoculation with Cauliflower mosaic virus (CaMV) in canola
varieties, A: without symptoms, B: mottling, C: vein clearing and D: mosaic and vein clearing.
)���2- Q�� )�H ����� DS� � ��NO B�$ PC�$ 9�8&���405 ��� ������� ����� ���R� ����" �/��&�& ��
)Cauliflower mosaic virus, CaMV(
Table2- Mean of symptoms severity index and absorbance at 405 nm wave length in ELISA of
Cauliflower mosaic virus (CaMV)
Cultivar No. of plants with score
3 and 4
Mean of severity index Mean of absorbance
(I) (M) (I) (M)
Hayola 401 8 3.00a 0g 2.83ab 1.23j
Zafar 4 2.08cde
0g 2.6
abcde 1.42
ij
Sary gol 6 2.50bc
0g 2.54
bcdef 1.33
ij
Karaj1 6 2.83ab
0g 2.17
fg 1.45
ij
Karaj2 4 2.08cde 0g 2.68abcd 1.31ij
Karaj3 0 1.42f 0g 2.55bcdef 1.18j
RGS003 4 2.50bc
0g 2.43
cdef 1.36
ij
Opera 0 1.83def
0g 2.28
efg 1.31
ij
Talayeh 0 1.92def 0g 2.51bcdef 1.57ij
Licord 2 2.00cde 0g 2.94a 1.36ij
Okapi 0 1.75def
0g 2.29
defg 1.71
hi
SLM046 0 1.58ef
0g 1.96
gh 1.42
ij
Zarfam 2 2.17cd
0g 2.79
abc 1.24
j
I : �� �!� �5� �� . CaMV
M : �� � �; �� . 8>. O\*>
H�<5�� I 7L% EF' �$ H�5%$ )0��+ 9'% 8. �.�� j�8L �%�%$ �5�61� %.�67*�5 �%$
I: Inoculated by CaMV,
M: Mock inoculation
Means values indicated by the same letters represent not significant differences at P ≤ 0.05 calculated using
the Duncan test.
A B
C D
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����� Z�IF � ��NO %�
Table3- Square means and significance level of ANOVA test in inoculated canola cultivars by
Cauliflower mosaic virus and its effect on symptoms and virus accumulation
Virus absorbance Mean of severity
index Degree of
freedom Source
638.545** 1156.049** 1 Virus 2.415** 4.309** 12 Genotype 5.157** 4.309** 12 Virus×Genotype
275 Error
312 Total
ns �61� j^7Y%:*�5 �%$
�61� j^7Y% * � ** I 7L% EF' �$ `�;8; �. �%$05/0 �01/0
Ns: not significant
*, **: Significant at the 0.05 and 0.01 probability levels, respectively
)���4- ����� � ���� ���� ����� ����� DS� � ��NO B�$ PC�$ (��4��^) �8/*W�� ����" V��/&
��� ������� ) ��Cauliflower mosaic virus, CaMV(
Table 4- Results of Pearson correlation test for symptoms severity index and virus absorbance in
canola cultivars reacted to Cauliflower mosaic virus.
Correlation coefficient Cultivar 0.909** Hayola 401 0.875** Zafar 0.672** Sary gol 0.644** Karaj1 0.896** Karaj2 0.919** Karaj3 0.892** RGS003 0.896** Opera 0.775** Talayeh 0.638** Licord 0.533** Okapi 0.533** SLM046 0.810** Zarfam 0.739** Total
I876� m!%8� �$ %��� Z(�% ,6�%� ��t( O1aF� �$
�!%�� 8.%8. �$ ����� � �� 9�8!� �% �%CaMV Z�5%
��:'%. Z ; �� $%$ )�5 �'�8. H!% �% TgL n!75
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�. :t*5 �87��. O�� . ,6�%� H!% Z(�% �Y8. �$ ��
��� Z(�% 8!') ��Walsh and Tomlinson,
1985; Walsh, 1989( �� ��5 8qL h�cK; �$ .
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9�8!�CaMV �� �'�8. �G Z(�% Z ; �� $%$ )�5
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Z(�% 8!' �% 87��. Z(�% �Y8. �$ &!^� O�� H�6o
Oc�cK; n!75 . �� $0.(1989) Walsh et al.
$�%$ :c.F�. �$ �%�!" :��c� )06�; ��6!% �. ��0; .
�$ �� H!% ]�;05[ 8.%8.CaMV �:'% ��5 ���
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CaMV �TuMV %��� )�� �% )%8!% �$ B�%��
:'% �� )Farzadfar and Pourrahim, 2014 p
Shahraeen, 2012 pTabarestani et al., 2010
(O�*Y �. H�6o � H!% �% ��5 ��$ �$ 9�8!�
:'% �� ��% ��5 %���)Walsh and Tomlinson,
1985; �Walsh et al., 1989; Coutts and
Jones, 2000;( 8.%8. �$ Z(�% H!% ,6�%� H!%8.6. �
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v8. �$ ����!� � . �� �5�TuMV ��5 B�%��
��:'% ) Jafari et al., 2016�% /" .( ��� :\
� �� S� 7*�' &!^� �$ v8. ��!��5� ��5 )��
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9�8!� �% S! 8 �% ��5 &!^� ��� �$ .��
&!^� ���aL CaMV $��L14 �!� �% /" ��� �5�
8� ��� &!^� TuMV �!� �% /" ��� :\ T.( �5�
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%8> %� %��� �� �:>8�. �% ��5 &!^� ���75 �$
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H�5%$ )0��+ �$ H�<5�� �*!c� n!75 .��5 ��� �!�.
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CaMV .$0. �.�� 9�8!� H!% . �gJ7Y% ��$0a+ �$
�6W 8 �% ��5 &!^� �0R� CaMV �% Z(�% �Y8. �$
��' �8\� T�t( P0! �T�401 f!8' Z>�� � �\;% 8;
O�� {Y� �%8. H�<5�� �*!c� n!75 .$7>%&!^�
TuMV ��$0a+ �$b0�-� �� $%$ )�5 9�8!� �$
�8� . �*!c� �$ Z(�% ��6. ,6�%� �'�8. n!75 Z(�%
. %���� TuMV $�%$ O�\; )Jafari et al., 2016( .
��$0a+ �$ H�<5�� b0�-� {Y�&!^� TuMV �$
��' &(� H�<5�� . T�83/6 I��� ) 5� ( ��$0a+ �$
. %��� Z(�%� TuMV ��;%��� &(�2 H�<5�� .66/7
,�. �'�8. �$ 8 �$ ���aL �$ :�%$ %� H�<5�� H!8;
u8� &(�3 �J7Y% $0Y �. %� &!^� {Y� H!87 �
$%$`�;8; . ,!��+ �$ H!% �$ Z(�% 8!' . �% O�\7� �
�8� 8<!��!�5�� ��6. I���)5 �Jafari et al.,
2016 .(�. ��6!% &V� �$�'�8. . %��� Z(�%� TuMV
u8� &(�2 �<a0;0� &!^� 75 �$ ���� ,�. H!8; ���$
H�<5�� %��:'% $%$ )�5 &!^� 8qL h�cK; �$
��%$8. :�%$$! �$ �<a0;0� .��5 :tk &(� H!% �%8.
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$ :t*5:�a1> �%8. �6�R. ��$ .$% TuMV $��L18
���$90�*�' �����. �. Z(�% ,6�%� �'�8.TuMV
�$ :'% �� Z�5% 9�8!� �6�R. ��$ �$�aL ���'�8.
��$ �$ 9�8!� �$ b0�-� ��$0a+22 90�*�' ���$
f �; )%��� ,6� 8. .�� Z�5%CaMV �TuMV �
�� �. O�� $%$ )�5 /�*"���.%�+ �� �$ R5+ �!%�
:'%�F.%� �50� }� ��& � 9�8!� �$ H!% )�� �!%�>%
)$�%�5 $0��Martín and Elena, 2009.(
79
������� �� ����)� ������ ��� �(39 � �3 ���!" 1395
)���5- ����� U��'� �� �!" ��NO B�$ PC�$ 9�8&������ ������� %��) ��Cauliflower mosaic virus,
CaMV ) �#�$ ������� � ( Turnip mosaic virus, TuMV(
Table 5- Mean of symptoms severity index in mixed infection by Cauliflower mosaic
virus and Turnip mosaic virus.
CaMV TuMV Cultivar
3.20a 5.17
f Hayola 401
2.12cde 6.33
bc Zafar
2.70ab 6.83a Sary gol
2.77ab 5.83
de Karaj1
2.15cde 6.50
ab Karaj2
1.30f 2.50
g Karaj3
2.40bc 6.00cd RGS003
1.88def 6.00
cd Opera
1.90def 6.00
cd Talayeh
2.00cde 5.83
de Licord
1.63ef 5.50ef Okapi
1.60ef 5.83
de SLM046
2.20cd 5.67
de Zarfam
H�<5�� I 7L% EF' �$ H�5%$ )0��+ 9'% 8. )07' 8 �$ �.�� j�8L �%�%$ �5 %�61�.�67*�5 �%$
Means values indicated by the same letters represent not significant differences at P ≤ 0.05 calculated using
the Duncan test.
�� ��5 8qL h�cK; n!75 :>8� Z�5% %��� �� ���
%� /�*"���.%�+ �� ��� �.�� n!75�� ��!;�6�.
&(� :�'*L �% ��L ,��" H!% n!75 �
:�� �%��� �. :t*5 )%8!% �$ ��CaMV ��$0a+ �
)�� CaMV �TuMV ��. �� �>8G �% � �$
)%8!% �$ %��� :�� hG6� `�V% �� H!% � �$8 �.
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9�8!� �. ��t*5 :��c� ��5 . ! � $0� n!�8; �5�%$
B�� �% $\7'%:-!�%8; S�75[ �'�6R� � ��'
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�%8( ��58. �$ )%8!%.$8��
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7
Plant Protection (Scientific Journal of Agriculture), 39(3) Fall 2016
Reaction of canola cultivars to Cauliflower mosaic virus and co-infection of
Turnip mosaic virus and Cauliflower mosaic virus under greenhouse condition
A. Ghasemzadeh1 and M. Shams-bakhsh
2*
1. Ph.D. Student, Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
2. *Corresponding Author: Associate Professor, Department of Plant Pathology, Faculty of Agriculture, Tarbiat
Modares University, Tehran, Iran ([email protected])
Received: 17 April 2016 Accepted: 22 October 2016
Abstract
The expanding cultivation of canola (Brassica napus L.) in the world indicates the importance of
this plant. Out of 12 reported viruses in canola, Cauliflower mosaic virus (CaMV) and Turnip
mosaic virus (TuMV) are of particular importance. Use of resistant cultivars would be the best
approach to control losses caused by viral diseases. So far, the reaction of commercial canola
cultivars in Iran to CaMV has not been assessed. In this study, 13 commercial varieties of canola
were evaluated against a CaMV isolate under greenhouse conditions. Analysis of symptom
severity index and enzyme-linked immunosorbent assay (ELISA) of inoculated plants showed
that Karaj3 cultivar was tolerant to this virus. In addition, due to co-infection of canola fields
with CaMV and TuMV, the reaction of commercial canola cultivars to mixed infections with
both viruses was evaluated. The obtained results showed that commercial canola cultivars in Iran
were susceptible to both CaMV and TuMV. However, Karaj3 cultivar was considered to be
moderately susceptible to CaMV and TuMV co-infection compared to other tested cultivars.
Key words: Brassica napus, ELISA, Resistance