scholarly journals Detection and Molecular Characterization of Watermelon Mosaic Virus (WMV) Spread Along the Syrian Coast

2021 ◽  
Vol 39 (1) ◽  
pp. 47-54
Author(s):  
Ahmad Mouhanna ◽  
◽  
Oos Ali Hasan ◽  
Hind Naaman Harhoush Al- Obaidi ◽  
◽  
...  

Mouhanna, A.M., A.A. Ali Hasan and H.N.H. Alobaidi. 2021. Detection and Molecular Characterization of Watermelon Mosaic Virus (WMV) Spread Along the Syrian Coast. Arab Journal of Plant Protection, 39(1): 47-54. Identification of local isolates of Watermelon mosaic virus (WMV) was investigated using serological and molecular methods. A total of 293 leaf samples from watermelon, zucchini, pumpkin, cucumber, pepper, beans and potato, with symptoms suggestive of virus infection, were collected from fields in two Syrian governorates (Latakia, Tartus) along the Syrian coast. DAS-ELISA tests revealed the presence of WMV in watermelon, zucchini, pumpkin and cucumber samples, with an average relative incidence of 36.95, 26.31, 29.27 and 37.70%, respectively. The infection of pepper with Watermelon mosaic virus was reported for the first time in Syria, with an average relative incidence of 2.94%. WMV was not detected in potato and beans, and these results were confirmed by RT-PCR. The local WMV isolate Cu4 was grouped with an Iranian isolate [EU660584.1] with 98.9% sequence identity. A Turkish isolate [EU660579] was grouped with local WMV isolate Wa2 with 98.3% sequence identity. Furthermore, two local isolates Zu6 and Cu8 represented one sub-group with 99.3% sequence identity. Keywords: Watermelon mosaic virus, DAS-ELISA, RT-PCR, CI, Phylogenetic Tree.

Genetika ◽  
2015 ◽  
Vol 47 (2) ◽  
pp. 651-663 ◽  
Author(s):  
Dragana Milosevic ◽  
Ivana Stankovic ◽  
Aleksandra Bulajic ◽  
Maja Ignjatov ◽  
Zorica Nikolic ◽  
...  

During 2009 and 2010, a survey was conducted in pepper crops to detect the possible presence of Pepper mild mottle virus (PMMoV) in Serbia. A total of 239 pepper samples from 39 crops at 26 localities were collected and analyzed for the presence of PMMoV, Cucumber mosaic virus (CMV), Potato virus Y (PVY), and Alfalfa mosaic virus (AMV), using DAS-ELISA test. Although it was detected in a small percentage, PMMoV could pose a threat to pepper production in Serbia due to its rapid seed-borne spread. Presence of PMMoV was confirmed by serological and biological detection, followed by conventional reverse transcription RT-PCR, using primers specific for the RNA-dependent RNA polymerase (RdRp) and the coat protein (CP) genes. Molecular identification confirmed that the Serbian isolates belong to PMMoV pathotypes P1,2 which do not break the resistance gene L3. Reconstructed phylogenetic tree confirmed the allocation of the Serbian isolates together with the majority of PMMoV isolates which belong to pathotypes P1,2. This study represents the first serological and molecular characterization of PMMoV infection of pepper in Serbia, and provides important data on the population structure. The obtained data could have great influence on pepper production in Serbia as well as future pepper resistance breeding in the country.


Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 702-702 ◽  
Author(s):  
I. Malandraki ◽  
N. Vassilakos ◽  
C. Xanthis ◽  
G. Kontosfiris ◽  
N. I. Katis ◽  
...  

In the summer of 2012, zucchini (Cucurbita pepo L.) plants of F1 hybrid Rigas showing very severe malformation and blisters in leaves and fruit were observed in the prefectures of Ilia and Messinia, Peloponnese, southwestern Greece. Over 100 samples were collected and only a few were found by double antibody sandwich (DAS)-ELISA to be singly or mixed infected with the commonly encountered Cucumber mosaic virus (CMV, genus Cucumovirus), Zucchini yellow mosaic virus (ZYMV, genus Potyvirus), and Watermelon mosaic virus (WMV, genus Potyvirus), to which Rigas is known to be tolerant. All affected plants were also tested by DAS-ELISA and RT-PCR (2) for the presence of Moroccan watermelon mosaic virus (MWMV; genus Potyvirus), a virus not previously reported in Greece, and were consistently found positive by both methods. Sap from plants in which MWMV was solely detected was used to mechanically inoculate Chenopodium quinoa Willd. and cucurbit species (zucchini, cucumber, melon, and watermelon). C. quinoa produced chlorotic local lesions, while cucurbits showed very severe mosaic and malformation of leaves. Zucchini plants of F1 hybrids Rigas, Golden (tolerant to WMV and ZYMV), and Elion (not exhibiting any tolerance) grown in a screenhouse produced equivalent severe symptoms on leaves and fruits. Furthermore, transmission experiments in a non-persistent manner using a clone of Myzus persicae Sulz. and zucchini plants of F1 hybrid Boreas as donor and test plants were carried out. Ten plants were used in each experiment (one aphid/plant) and this was repeated five times (50 plants in total). The transmission rate was high ranging from 75 to 90%. RT-PCR obtained amplicons of 627 bp were subjected to direct sequencing (GenBank Accession No KF772944), which revealed 99% sequence identity to the corresponding region of a MWMV Tunisian isolate (EF579955). In 2013, in addition to zucchini plants found MWMV positive, watermelon (Citrullus lanatus Thunb.) plants from the same region of Peloponnese showing leaf malformation and mosaic symptoms were found MWMV positive (4/30) by DAS-ELISA and RT-PCR, revealing the virus establishment and further spread. In the Mediterranean basin, the virus has already been reported in Morocco, Italy, France, Spain, Tunisia, and Algeria, where it has emerged recently from a common source, has quickly become established through rapid dissemination and is considered as an important emerging threat (4). Isolates from these countries, including the present one from Greece, are very closely molecularly related to each other, contrary to isolates from sub-Saharan Africa (South Africa, Sudan, Congo, Zimbabwe, Niger, Cameroon, Nigeria) that are much more divergent (1,3). To our knowledge, this is the first report of MWMV in Greece. References: (1) H. Lecoq et al. Plant Dis. 85:547, 2001. (2) H. Lecoq et al. New Dis. Rep. 16:19, 2007. (3) A. T. Owolabi et al. Int. J. Virol. 8:258, 2012. (4) S. Yakoubi et al. Arch. Virol. 153:775, 2008.


2004 ◽  
Vol 29 (2) ◽  
pp. 215-219 ◽  
Author(s):  
Manuela R. Barbieri ◽  
Murilo G. de Carvalho ◽  
Eunize M. Zambolim ◽  
F. Murilo Zerbini

Um anti-soro policlonal específico para Watermelon mosaic virus (WMV) foi produzido por meio da imunização de coelhos com proteína capsidial purificada, expressa in vitro em células de Escherichia coli. O gene cp foi amplificado via RT-PCR utilizando oligonucleotídeos específicos, a partir de RNA viral extraído de preparações virais concentradas. O fragmento amplificado foi clonado em pBLUESCRIPT KS+ e completamente seqüenciado para confirmação de sua identidade e integridade. Em seguida, o fragmento foi subclonado no vetor de expressão pRSET-A. Plasmídeos recombinantes foram utilizados para a expressão da proteína capsidial em E. coli BL21::DE3. A proteína foi purificada por meio de cromatografia de afinidade em coluna de Ni+-NTA, a partir de proteínas totais extraídas de E. coli. Uma vez purificada, a proteína foi quantificada e utilizada para imunização dos coelhos. O anti-soro foi testado quanto a sua especificidade e sensibilidade em testes de Western blot, DAS-ELISA, imunodifusão e ELISA indireto. Todos os testes demonstraram que o anti-soro produzido a partir da expressão in vitro da proteína capsidial é altamente específico para a detecção do WMV em extratos foliares, não tendo sido observada nenhuma reação heteróloga interespecífica.


2011 ◽  
Vol 132 (3) ◽  
pp. 317-322 ◽  
Author(s):  
Mariella Matilde Finetti-Sialer ◽  
Tiziana Mascia ◽  
Fabrizio Cillo ◽  
Crisostomo Vovlas ◽  
Donato Gallitelli

2021 ◽  
Vol 26 (51) ◽  
pp. 33-39
Author(s):  
Dragana Milošević ◽  
Maja Ignjatov ◽  
Zorica Nikolić ◽  
Gordana Tamindžić ◽  
Gordana Petrović ◽  
...  

In the 2018-19 growing season, a total of 51 leaves of eggplant plants grown under field conditions were collected randomly from nine private gardens at four different localities in the Province of Vojvodina. Eggplants with nearly 40% of plants showing bright yellow to white mosaic or mottling of leaves were found throughout the inspected fields (gardens). The collected samples were analyzed for the presence of alfalfa mosaic virus (AMV), cucumber mosaic virus (CMV) and potato virus Y (PVY) using commercial double-antibody sandwich (DAS)-ELISA kits. Serological analysis of eggplant samples revealed the presence of AMV in 80.39% collected samples. None of the analyzed samples was positive for CMV and PVY. The virus was successfully mechanically transmitted to test plants including Nicotiana benthamiana, Chenopodium quinoa, C. amaranticolor, as well as eggplant seedlings, confirming the infectious nature of the disease. The presence of AMV in eggplants was further verified by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing, using the primers CP AMV1 and CP AMV2 that amplify part of the coat protein (CP) gene. The phylogenetic analysis showed that Serbian AMV isolates grouped into a separate well-supported group together with AMV isolates from Italy, Croatia and previously characterized isolates from Serbia. To our knowledge, this is the first report of AMV infection of eggplant in Serbia.


Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1254
Author(s):  
Hang Yin ◽  
Zheng Dong ◽  
Xulong Wang ◽  
Shuhao Lu ◽  
Fei Xia ◽  
...  

Marigold plants with symptoms of mosaic, crinkle, leaf curl and necrosis were observed and small RNA and ribo-depleted total RNA deep sequencing were conducted to identify the associated viruses. Broad bean wilt virus 2, cucumber mosaic virus, turnip mosaic virus, a new potyvirus tentatively named marigold mosaic virus (MMV) and a new partitivirus named as marigold cryptic virus (MCV) were finally identified. Complete genome sequence analysis showed MMV was 9811 nt in length, encoding a large polyprotein with highest aa sequence identity (57%) with the putative potyvirus polygonatumkingianum virus 1. Phylogenetic analysis with the definite potyviruses based on the polyprotein sequence showed MMV clustered closest to plum pox virus. The complete genome of MCV comprised of dsRNA1 (1583 bp) and dsRNA2 (1459 bp), encoding the RNA-dependent RNA polymerase (RdRp), and coat protein (CP), respectively. MCV RdRp shared the highest (75.7%) aa sequence identity with the unclassified partitivirus ambrosia cryptic virus 2, and 59.0%, 57.1%, 56.1%, 54.5% and 33.7% with the corresponding region of the definite delta-partitiviruses, pepper cryptic virus 2, beet cryptic virus 3, beet cryptic virus 2, pepper cryptic virus 1 and fig cryptic virus, respectively. Phylogenetic analysis based on the RdRp aa sequence showed MCV clustered into the delta-partitivirus group. These findings enriched our knowledge of viruses infecting marigold, but the association of the observed symptom and the identified viruses and the biological characterization of the new viruses should be further investigated.


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