The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine
Aim. In current work, we proceeded with the strain attribution of Ukrainian isolates CMV based on the phylogenetic analysis of the partial sequences of the coat protein gene. Methods. ELISA, RT-PCR, DNA sequencing and phylogenetic analysis. Results. Cucumber mosaic virus (CMV) is widespread among th...
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| Zitieren: | The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine / T.P. Shevchenko, O.V. Tymchyshyn, E. AlDalain, A.S. Bysov, I.G. Budzanivska, O.V. Shevchenko, V.P. Polishchuk // Biopolymers and Cell. — 2015. — Т. 31, № 1. — С. 57-62. — Бібліогр.: 15 назв. — англ. |
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nasplib_isofts_kiev_ua-123456789-1524352025-02-09T17:55:59Z The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine Перше повідомлення про детекцію ізолятів підгрупи IB вірусу огіркової мозаїки в Україні Первое сообщение о детекции изолятов подгруппы IB вируса мозаики огурца в Украине Shevchenko, T.P. Tymchyshyn, O.V. AlDalain, E. Bysov, A.S. Budzanivska, I.G. Shevchenko, O.V. Polishchuk, V.P. Viruses and Cell Aim. In current work, we proceeded with the strain attribution of Ukrainian isolates CMV based on the phylogenetic analysis of the partial sequences of the coat protein gene. Methods. ELISA, RT-PCR, DNA sequencing and phylogenetic analysis. Results. Cucumber mosaic virus (CMV) is widespread among the variety of crops from the Cucurbitaceae and Solanaceae families in Ukraine. The symptomatic samples from different regions of Ukraine were collected and tested for the presence of CMV. The coat protein (CP) gene of two isolates was amplified and sequenced. The partial nucleotide sequences of CP gene were determined and compared to those of other CMV strains belonging to the IA, IB and II subgroups. Comparison of the nucleotide sequences of Ukrainian isolates showed their similar identity percentages and close relationships with the subgroup IB strains from other countries. The highest nucleotide homology was shared with the strains ABI (Korea) and SD (China). Conclusions. Based on the highest identities of the coat protein gene sequences and close phylogenetic relationships with the subgroup IB members of CMV, the Ukrainian isolates under study were identified as belonging to the subgroup IB. Our findings show for the first time an occurrence of the IB subgroup isolates of CMV in Ukraine. Мета. Встановлення штамової приналежності українських ізолятів вірусу огіркової мозаїки (ВОМ) на основі філогенетичного аналізу фрагмента послідовності гена капсидного білка. Методи. Імуноферментний аналіз (ІФА), полімеразна ланцюгова реакція зі зворотною транскрипцією (ЗТ-ПЛР), сиквенування ДНК та філогенетичний аналіз. Результати. ВОМ є широко розповсюдженим патогеном сільськогосподарських культур в Україні. Були відібрані й проаналізовані зразки рослин з вірусоподібними симптомами представників родин Cucurbitaceae і Solanaceae. Виходячи з результатів візуальної та серологічної діагностики, два зразки Lycopersicon esculentum і Cucurbita pepo з Полтавської області були обрані для подальших молекулярних досліджень. Були ампліфіковані й сиквеновані часткові послідовності гена капсидного білка. Отримані послідовності кДНК гена капсидного білка цих ізолятів порівняли з опублікованими в Генбанку послідовностями штамів ВОМ різних підгруп. Найбільша гомологія українських ізолятів продемонстрована зі штамами ВОМ підгрупи ІВ. Найбільш спорідненими до українських ізолятів виявилися штам АВІ з Кореї та штам SD з Китаю. Висновки. Спираючись на результати філогенетичного аналізу, можна стверджувати, що детектовані українські ізоляти вірусу огіркової мозаїки належать до групи ІВ. Наші результати є першим повідомленням про наявність ізолятів ВОМ підгрупи ІВ на території України. Цель. Установить штаммовую принадлежности украинских изолятов вируса мозаики огурца (ВМО) на основании филогенетичного анализа частичной последовательности гена капсидного белка. Методы. Иммуноферментный анализ (ИФА), полимеразная цепная реакция с обратной транскрипцией (ОТ-ПЦР), секвенирование ДНК и филогенетический анализ. Результаты. ВМО широко распространён в Украине среди сельскохозяйственных культур. Были отобраны и проанализированы образцы растений с вирусоподобными симптомами семейств Cucurbitaceae и Solanaceae. На основе визуальной и серологической диагностики два образца Lycopersicon esculentum и Cucurbita pepo из Полтавской области были избраны для дальнейших молекулярных исследований. Были амплифицированы и секвенированы частичные последовательности гена капсидного белка. Проведено сравнение полученных последовательностей кДНК гена капсидного белка этих изолятов с опубликованными в Генбанке последовательностями штаммов ВМО различных подгрупп. Наибольшая гомология украинских изолятов продемонстрирована с представителями подгруппы ІВ. Наиболее родственными к украинским изолятам показаны штаммы АВІ (Корея) и SD (Китай). Выводы. Исходя из результатов филогенетического анализа, украинские изоляты ВОМ принадлежат к подгруппе ІВ. Результаты этой работы являются первым свидетельством присутствия изолятов подгруппы ІВ ВМО на территории Украины. 2015 Article The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine / T.P. Shevchenko, O.V. Tymchyshyn, E. AlDalain, A.S. Bysov, I.G. Budzanivska, O.V. Shevchenko, V.P. Polishchuk // Biopolymers and Cell. — 2015. — Т. 31, № 1. — С. 57-62. — Бібліогр.: 15 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.0008CD https://nasplib.isofts.kiev.ua/handle/123456789/152435 578.856 en application/pdf |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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Viruses and Cell Viruses and Cell |
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Viruses and Cell Viruses and Cell Shevchenko, T.P. Tymchyshyn, O.V. AlDalain, E. Bysov, A.S. Budzanivska, I.G. Shevchenko, O.V. Polishchuk, V.P. The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine |
| description |
Aim. In current work, we proceeded with the strain attribution of Ukrainian isolates CMV based on the phylogenetic analysis of the partial sequences of the coat protein gene. Methods. ELISA, RT-PCR, DNA sequencing and phylogenetic analysis. Results. Cucumber mosaic virus (CMV) is widespread among the variety of crops from the Cucurbitaceae and Solanaceae families in Ukraine. The symptomatic samples from different regions of Ukraine were collected and tested for the presence of CMV. The coat protein (CP) gene of two isolates was amplified and sequenced. The partial nucleotide sequences of CP gene were determined and compared to those of other CMV strains belonging to the IA, IB and II subgroups. Comparison of the nucleotide sequences of Ukrainian isolates showed their similar identity percentages and close relationships with the subgroup IB strains from other countries. The highest nucleotide homology was shared with the strains ABI (Korea) and SD (China). Conclusions. Based on the highest identities of the coat protein gene sequences and close phylogenetic relationships with the subgroup IB members of CMV, the Ukrainian isolates under study were identified as belonging to the subgroup IB. Our findings show for the first time an occurrence of the IB subgroup isolates of CMV in Ukraine. |
| format |
Article |
| author |
Shevchenko, T.P. Tymchyshyn, O.V. AlDalain, E. Bysov, A.S. Budzanivska, I.G. Shevchenko, O.V. Polishchuk, V.P. |
| author_facet |
Shevchenko, T.P. Tymchyshyn, O.V. AlDalain, E. Bysov, A.S. Budzanivska, I.G. Shevchenko, O.V. Polishchuk, V.P. |
| author_sort |
Shevchenko, T.P. |
| title |
The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine |
| title_short |
The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine |
| title_full |
The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine |
| title_fullStr |
The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine |
| title_full_unstemmed |
The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine |
| title_sort |
first evidence of subgroup ib isolates of cucumber mosaic virus in ukraine |
| publishDate |
2015 |
| topic_facet |
Viruses and Cell |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/152435 |
| citation_txt |
The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine / T.P. Shevchenko, O.V. Tymchyshyn, E. AlDalain, A.S. Bysov, I.G. Budzanivska, O.V. Shevchenko, V.P. Polishchuk // Biopolymers and Cell. — 2015. — Т. 31, № 1. — С. 57-62. — Бібліогр.: 15 назв. — англ. |
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57
Viruses and Cell
UDC 578.856
The fi rst evidence of subgroup IB isolates
of Cucumber mosaic virus in Ukraine
T. P. Shevchenko, O. V. Tymchyshyn, E. AlDalain,
A. S. Bysov, I. G. Budzanivska, O. V. Shevchenko, V. P. Polishchuk
Educational and Scientifi c Center «Institute of Biology»,
Taras Shevchenko National University of Kyiv
64/13, Volodymyrska Str., Kyiv, Ukraine, 01601
tyvonchuk@ukr.net
Aim. In current work, we proceeded with the strain attribution of Ukrainian isolates CMV based on the
phylogenetic analysis of the partial sequences of the coat protein gene. Methods. ELISA, RT-PCR, DNA
sequencing and phylogenetic analysis. Results. Cucumber mosaic virus (CMV) is widespread among the
variety of crops from the Cucurbitaceae and Solanaceae families in Ukraine. The symptomatic samples
from different regions of Ukraine were collected and tested for the presence of CMV. The coat protein (CP)
gene of two isolates was amplifi ed and sequenced. The partial nucleotide sequences of CP gene were deter-
mined and compared to those of other CMV strains belonging to the IA, IB and II subgroups. Comparison
of the nucleotide sequences of Ukrainian isolates showed their similar identity percentages and close rela-
tionships with the subgroup IB strains from other countries. The highest nucleotide homology was shared
with the strains ABI (Korea) and SD (China). Conclusions. Based on the highest identities of the coat pro-
tein gene sequences and close phylogenetic relationships with the subgroup IB members of CMV, the
Ukrainian isolates under study were identifi ed as belonging to the subgroup IB. Our fi ndings show for the
fi rst time an occurrence of the IB subgroup isolates of CMV in Ukraine.
K e y w o r d s: Cucumber mosaic virus, coat protein gene, phylogenetic analysis.
Introduction
Cucumber mosaic virus (CMV) is a type species
from the genus Cucumovirus, family Bromoviridae.
It is distributed worldwide in temperate and tropical
areas causing epidemics in a variety of economically
important crops. CMV is able to infect approximate-
ly 1300 species of more than 500 mono- and dicoty-
ledonous plant genera among over 100 families, with
new hosts reported every year [1]. Wide dissemina-
tion of CMV suggests its success in rapid adapting to
new hosts and new environments [2].
The genome of CMV is a single-stranded, positive-
sense RNA. There are three RNA segments contain-
ing fi ve open reading frames (ORF), which code for
the proteins 1a, 2a, 2b, 3a, and coat protein (CP) [3].
RNAs 1 and 2 encode the components of the viral
polymerase complex (1a and 2a). RNA 2 also encodes
a protein functioning as a suppressor of the posttran-
scriptional gene silencing (2b) [4]. RNA 3 contains
the 3a and coat protein genes separated by an inter-
genic region. Both of these genes are necessary for
cell-to-cell movement of the virus in the host plant
[3]. The 3a protein corresponds to the movement pro-
tein (MP). The coat protein is expressed from a subge-
nomic RNA (RNA 4) and plays an important role in
aphid transmission and symptom expression [5, 6].
According to serological relationships, peptide
mapping of the coat protein, nucleic acid hybridiza-
tion and nucleotide sequence identity, a number of
CMV isolates have been described previously and
classifi ed into two subgroups, designated I and II
ISSN 0233-7657
Biopolymers and Cell. 2015. Vol. 31. N 1. P. 57–62
doi: http://dx.doi.org/10.7124/bc.0008CD
© 2015 T. P. Shevchenko et al.; Published by the Institute of Molecular Biology and Genetics, NAS of Ukraine on behalf of Biopolymers and Cell.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/),
which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited
58
T. P. Shevchenko, O. V. Tymchyshyn, E. AlDalain et al.
[7]. The development of phylogenetic analysis meth-
ods led to the further subdivision of CMV isolates of
subgroup I into subgroups IA and IB [2]. The phy-
logeny estimations with full CP ORF, as well as the
rearrangements in the 5’ nontranslated region of
RNA 3 verifi ed these designations [8].
The strains belonging to subgroups I and II have
dissimilar characteristics. The CMV strains of sub-
group I are considered more virulent than those of
subgroup II. The members of subgroup IA occur all
over the world. Contrarily, most isolates in subgroup
IB are reported from East Asia, which is considered
to be the origin of this subgroup [9]. The strains and
isolates detected in other areas (the Mediterranean
region, California, Brazil, Australia) have been in-
troduced recently from Asia [1]. The mild strains be-
long to subgroup II; they are widespread in cooler
areas of temperate regions, in the USA, Australia,
and Africa [1].
Previously, CMV of subgroup II has been detected
in infected pumpkin plants from Ukraine [10]. The
subgroup attribution has been established based on
the RT-PCR product size. However, the sequences of
these isolates have not been obtained and their com-
parison with other known strains and isolates has not
been performed. In current work, we proceeded with
the strain attribution of Ukrainian isolates based on
phylogenetic analysis of the partial sequences of the
coat protein gene.
Materials and Methods
Plant samples were collected from different regions
of Ukraine. The symptomatic samples were scree ned
for the presence of viral antigens. Double-antibody
sandwich enzyme linked immunosorbent assay (DAS-
ELISA) was conducted using commercial test-system
of Loewe (Germany). Plant ma terial was homoge-
nized in 0.1M phosphate buffered saline (PBS), pH
7.4, 1:2 (m/v). Plant com ponents were removed by
centrifugation at 5.000 g for 20 min at +4 C using
centrifuge PC-6. The supernatant was taken for fur-
ther ELISA. DAS-ELISA was performed according
to the manufacturer’s recommendations. The results
were checked at the wavelength of 405/630 nm us-
ing microplate reader Termo Labsystems Opsis MR
(USA) with software Dynex Revelation Quicklink [11].
Total RNA was extracted from naturally infected
plant samples using RNeasy Plant Mini kit (Qia gen,
UK). The results were confi rmed by electrophoresis
of nucleic acids in 1.5% agarose gel. The two-step
reverse transcription reaction (RT-PCR) was accom-
plished using two specifi c primers complementary to
coat protein gene of CMV producing the amplicon
with expected size of 500 bp [12]: forward primer –
5’ TATGATAAGAAGCTTG TTTCGCGCA-3΄; re-
verse primer – 5’ TTTTAG CCGTAAGCTGGATGG
ACAACCC-3΄.
The fi rst strand cDNA synthesis was performed at
42 C for 60 min using Termo Scientifi c RevertAid
Reverse Transcriptase according to the manufactur-
er’s instructions. The fi rst cycle was carried out at 95
C for 1 min. PCR was conducted for 30 cycles us-
Fig. 1. Viral disease symptoms on plants under fi eld conditions
induced by Cucumber mosaic virus: A) discoloration of tomato
fruit; B) dark green mosaic of leaf blade on squash
A
B
59
The fi rst evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine
ing the following parameters: 30 s at 95 C, 30 s at
56 C, 1 min at 72 C, and then 5 min at 72 C in the
fi nal cycles. PCR amplifi cation was assessed by elec-
trophoresis in a 1.5 % agarose gel in TBE buffer (89
mM TRIS borate and 2 mM EDTA, pH 8.3) and
stained in ethidium bromide. The purifi ed amplicons
were sequenced using Applied Biosystems 3730x1
DNA Analyzer with Big Dye terminators, version
3.1 (Applied Biosystems, USA).
The aligned sequences of the part of coat protein gene
of two CMV isolates were compared with the published
sequences of CMV strains belonging to different sub-
groups available in the GenBank database using NCBI/
BLAST (http://www.ncbi.nlm.nih. gov/). The list of 20
CMV strains from NCBI used for phylogenetic analysis
is presented in Table 1. The phylogenetic analysis was
conducted using ME GA version 5.2.1 software. A phy-
logenetic tree was constructed using the Neighborhood
Joining method. The Kimura 2-parameter model was
used to estimate the nucleotide distances among the
compared sequences of the CP gene [13].
Results and Discussion
The symptomatic samples of vegetable plants were
observed under fi eld conditions in different loca-
tions. 126 plant samples belonging to the Cucurbita-
ceae and Solanaceae families were selected and
tested for CMV. The plant samples were collected
from following regions of Ukraine: Autonomic Re-
public of Crimea, Vinnytsia, Zaporizhzhia, Ky iv,
Kirovohrad, Odessa, Poltava, Cherkasy and Cherni-
hiv regions. The plants of Cucurbitaceae family
Fig. 2. Phylogenetic tree based on partial coat protein gene sequence of selected strains and Ukrainian isolates. Bootstrap values are
shown above branches
60
T. P. Shevchenko, O. V. Tymchyshyn, E. AlDalain et al.
(cucumber, squash, pumpkin, and zucchini) showed
puckering, distortion, vein banding, yellowing, fi la-
mentary leaf structures, yellow leaf mosaics; dark
green spots of different size, knobs and malforma-
tions on fruits. The plants of Solanaceae family (to-
mato, pepper, eggplant) showed disease symptoms
in one month after seedtime du ring fl owering. At
fi rst yellow spots and vein clearing have appeared on
young leaves, followed by systemic yellow and
green mosaics, chloroses and necroses. DAS-ELISA
was performed for detection of viral antigens in plant
samples. 38 of 126 plant samples were found CMV-
positive. Virus-infected plants were detected in ag-
riecosystems of Vinnytsia, Zaporizhzhia, Kyiv, Odes-
sa, Poltava and Cherkasy regions.
For further investigations, two samples were cho sen
since they showed a high virus titer in DAS-ELISA and
exclusive occurrence of CMV (among the ran ge of vi-
ruses tested). The tomato (Lycopersicon esculentum)
from Poltava region showed yellow spot ting on fruits
and leaf rolling. Such manifestation is considered classi-
cal symptoms of CMV with low rate of occurrence in
Ukraine (Fig. 1). The source of another isolate was
squash (Cucurbita pepo) from Poltava region with dark
green mosaic along the leaf veins (Fig. 1).
RT-PCR with the extracted total RNA resulted in am-
plifi cation of a cDNA (expected 500 bp long) covering a
part of CP gene. The partial CP gene nucleotide sequenc-
es of Ukrainian CMV isolates were obtained.
A phylogenetic tree was constructed using ali gned
nucleotide sequences of the partial CP gene of various
CMV strains isolated from different co untries. The
Phylogenetic analysis indicated three distinct clusters
corresponding to subgroups IA, IB and II (Fig. 2).
Table 1. Strains of Cucumber mosaic virus used for phylogenetic analysis in this work
Strain Sub
group Host Origin Accession
number
Nucleotide homology
Ukr-sq13 Ukr-tom2
237 IA Cucumis melo Australia AJ585518 94% 94%
ABI IB Korea L36525 98% 98%
ALS II Host – Alstroemeria, lab host –
Nicotiana benthamiana
Netherlands AJ276587 79% 79%
banana IA Musa acuminata Israel U43888 94% 94%
G10 IB Tobacco Greece AY541691 95% 95%
G2 IB Tobacco Greece AY450854 95% 95%
Hnt II Tobacco China KC407999 81% 81%
I17F IA France Y18137 94% 94%
K IB USA AF127977 98% 97%
LS II USA AF127976 82% 82%
N IA Limonium sinuatum Japan D28486 95% 95%
R II France Y18138 82% 82%
Rb IA Rudbeckia hirta var. Pulcherrima South Korea GU327365 95% 95%
S II Lycopersicon esculentum USA AF172841 81% 81%
SD IB China AB008777 99% 99%
TN II From tomato plants, lab host –
Nicotiana tabacum
Japan AB176847 80% 80%
Twa IA Pepper Ausralia AJ585522 95% 95%
V IA Lab host – Nicotiana benthamiana South Korea AB369270 94% 94%
Vir IB Capsicum sp. Italy HE962480 95% 95%
Z IA Lab host –Nicotiana benthamiana South Korea AB369269 95% 95%
61
The fi rst evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine
The CMV strains belonging to subgroup II formed
a different cluster on the phylogenetic tree, well sepa-
rated from the members of subgroup I. Within sub-
group I, Ukrainian CMV isolates sho wed close phylo-
genetic relationships rather with the members of sub-
group IB than IA, which formed a separate cluster.
Ukrainian isolates shared 79–99 % nucleotide ho-
mology with the strains reported from all over the world
(Table 1). According to literature data, the members of
the same group share more than 90 % homology. The
homology between strains from I and II subgroups was
approximately 69–77 % [3]. The nucleotide sequences
shared 92–94 % similarity among IA and IB subgroup
strains [14]. Ukrainian isolates were phylogenetically
most related to each other and to the members of sub-
group IB (>95 % nucleotide homology). However, they
were distinct from the subgroup IA and subgroup II
strains. The members of subgroup II showed the lowest
nucleotide homology (79–82 %) with the Ukrainian
isolates of CMV studied.
The phylogenetic analysis of partial sequences of
the CP gene of Ukrainian isolates of CMV revealed
the highest homology and close relationships with the
strains ABI and SD from Korea and China. They
shared approximately 98–99 % homology. The infre-
quent amino acid substitutions revealed a high simi-
larity in this gene region. The protein sequence of ABI
strain is different from those of Ukr-sq13, Ukr-tom2,
and SD, and has two amino acid substitutions (YA –
IT corresponding to 601, 602 positions). The sequence
comparison of Ukrainian isolates showed their high
similarity. Based on the results obtained, these Ukrai-
nian isolates of CMV were identifi ed to belong to the
subgroup IB. The sequence data have been submitted
to NCBI, accession numbers KJ921838 and KJ921837
for Ukr-tom2 and Ukr-sq13 isolates, respectively.
Conclusions
The results obtained confi rm the wide dissemination
of CMV in Ukraine. The analysis of many samples
collected from nine regions of Ukraine representing
approximately 1/3 of the country area suggests that
about 30 % of plants with manifested symptoms
were CMV-positive. Despite the supposed high rate
of CMV evolution and adaptation [2], the phyloge-
netic analysis of two Ukrainian isolates of CMV re-
veals that their homology exceeds 95 %, i.e. that the-
se isolates belong to the same strain. In our opinion,
it is of special interest as these isolates have been
found in totally different host plants, tomato and
squash, which belong to distant families.
Another important phylogenetic fi nding is that
Ukrainian isolates of CMV are attributed to the sub-
group IB of CMV strains. The Ukrainian isolates are
mostly related to the strains ABI and SD from Korea
and China, respectively. Initially, the members of sub-
group IB were found in (and thought to be restricted to)
the East Asia. Later on, the subgroup IB strains of CMV
were shown to be widespread in Iran [1], however the
subgroup IA isolates were also detected [15]. The oc-
currence of subgroup IB isolates of CMV suggests that
they might have been introduced in Ukraine either by
seed material or through the exported fresh food prod-
ucts. Additionally, we cannot reject a possibility of the
virus ‘natural migration’ from Iran (or other neighbor-
ing regions) to Ukraine by aphid or birds.
Strangely, we have not found the subgroup IA isolates
of CMV in Ukraine as yet. These are more virulent strains
and isolates which are considered to be common world-
wide, including Europe. In accordance with Zi ti kaitė I, et
al. [10], the CMV subgroup II was detected in pumpkins
from Ternopol and Chernivtsi regions of Uk rai ne. Thus,
based on previous research data and our fi n dings, the
CMV population in Ukraine consists of the isolates of II
and IB subgroups. Having registered severe symptoms
on collected plants, we expected that they were induced
by a virulent ‘form’ of the virus. Surprisingly, both iso-
lates fall into the subgroup IB strains of CMV. In the view
of aforesaid, we deem that the obtained results rather re-
fl ect the lack of CMV monitoring in Ukraine than reveal
atypical virus spread in the region.
The data also indicate that the severity of the vi-
rus-specifi c symptoms in fi eld conditions may not be
directly related to the degree of virulence of a given
virus isolate, requiring more research on the CMV
biology, epidemiology and evolution.
Acknowledgements
The authors are grateful to Dr. I. I. Boubriak for his
invaluable support and technical assistance.
62
T. P. Shevchenko, O. V. Tymchyshyn, E. AlDalain et al.
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logical and molecular characterization of Cucumber mosaic
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ber. Indian J Virol. 2013;24(1):27-34.
15. Nematollahi S, Sokhandan-Bashir N, Rakhshandehroo F, Za-
manizadeh HR. Phylogenetic analysis of new isolates of
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genomic regions. Plant Pathol J. 2012; 28(4): 381-9.
Т. П. Шевченко, О. В. Тимчишин, Е. Аль Далаін,
А. С. Бисов, І. Г. Будзанівська, О. В. Шевченко, В. П. Поліщук
Перше повідомлення про детекцію ізолятів
підгрупи IB вірусу огіркової мозаїки в Україні
Мета. Встановлення штамової приналежності українських ізо-
лятів вірусу огіркової мозаїки (ВОМ) на основі філогенетично-
го аналізу фрагмента послідовності гена капсидного білка. Ме-
тоди. Імуноферментний аналіз (ІФА), полімеразна ланцюгова
реакція зі зворотною транскрипцією (ЗТ-ПЛР), сиквенування
ДНК та філогенетичний аналіз. Результати. ВОМ є широко
розповсюдженим патогеном сільськогосподарських культур в
Україні. Були відібрані й проаналізовані зразки рослин з вірусо-
подібними симптомами представників родин Cucurbitaceae і
Solanaceae. Виходячи з результатів візуальної та серологічної
діагностики, два зразки Lycopersicon esculentum і Cucurbita pepo
з Полтавської області були обрані для подальших молекуляр-
них досліджень. Були ампліфіковані й сиквеновані часткові по-
слідовності гена капсидного білка. Отримані послідовності
кДНК гена капсидного білка цих ізолятів порівняли з опубліко-
ваними в Генбанку послідовностями штамів ВОМ різних під-
груп. Найбільша гомологія українських ізолятів продемонстро-
вана зі штамами ВОМ підгрупи ІВ. Найбільш спорідненими до
українських ізолятів виявилися штам АВІ з Кореї та штам SD з
Китаю. Висновки. Спираючись на результати філогенетичного
аналізу, можна стверджувати, що де тек товані українські ізоляти
вірусу огіркової мозаїки належать до групи ІВ. Наші результати
є першим по відомленням про наявність ізолятів ВОМ підгрупи
ІВ на території України.
Ключов і слова: Вірус огіркової мозаїки, ген кап сид но-
го білку, філогенетичний аналіз.
Т. П. Шевченко, О. В. Тымчишин, Е. Аль Далаин,
А. С. Бысов, И. Г. Будзанивская, А. В. Шевченко, В. П. Полищук
Первое сообщение о детекции изолятов
подгруппы IB вируса мозаики огурца в Украине
Цель. Установить штаммовую принадлежности украинских
изолятов вируса мозаики огурца (ВМО) на основании филоге-
нетичного анализа частичной последовательности гена капсид-
ного белка. Методы. Иммуноферментный анализ (ИФА), поли-
меразная цепная реакция с обратной транскрипцией (ОТ-ПЦР),
секвенирование ДНК и филогенетический анализ. Результаты.
ВМО широко распространён в Украине среди сельскохозяйс-
твенных культур. Были отобраны и проанализированы образцы
растений с вирусоподобными симптомами семейств Cu cur-
bitaceae и Solanaceae. На основе визуальной и серологической
диагностики два образца Lycopersicon esculentum и Cucurbita
pepo из Полтавской области были избраны для дальнейших мо-
лекулярных исследований. Были амплифицированы и секвени-
рованы частичные последовательности гена капсидного белка.
Проведено сравнение полученных последовательностей кДНК
гена капсидного белка этих изолятов с опубликованными в Ген-
банке последовательностями штам мов ВМО различных под-
групп. Наибольшая гомология украинских изолятов проде-
монстрирована с представителями подгруппы ІВ. Наиболее
родственными к украинским изолятам показаны штаммы АВІ
(Корея) и SD (Китай). Выводы. Исходя из результатов филоге-
нетического анализа, украинские изоляты ВОМ принадлежат к
подгруппе ІВ. Результаты этой работы являются первым свиде-
тельством присутствия изолятов подгруппы ІВ ВМО на терри-
тории Украины.
Ключевые слова: вирус мозаики огурца, ген кап сид-
ного белка, филогенетический анализ.
Received 24.12.2014
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