scholarly journals Simultaneous detection of watermelon mosaic virus (WMV) and zucchini yellow mosaic virus (ZYMV) using duplex RT-PCR reaction

2021 ◽  
Vol 61 (1) ◽  
Keyword(s):  
Mosaic Virus ◽  
Zucchini Yellow Mosaic Virus ◽  
Simultaneous Detection ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Rt Pcr

scholarly journals First Report of Papaya ringspot virus and Zucchini yellow mosaic virus in Luohanguo (Siraitia grosvenorii) in China

Plant Disease ◽  
2005 ◽  
Vol 89 (5) ◽  
pp. 530-530 ◽  
Author(s):  
Y.-M. Liao ◽  
X.-J. Gan ◽  
B. Chen ◽  
J.-H. Cai
Keyword(s):  
Mosaic Virus ◽  
Zucchini Yellow Mosaic Virus ◽  
Viral Disease ◽  
Ringspot Virus ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Papaya Ringspot Virus ◽  
Rt Pcr ◽  
First Report ◽  
Siraitia Grosvenorii

Luohanguo, Siraitia grosvenorii (Swingle) C. Jeffrey, is a perennial cucurbitaceous plant that is an economically important medicinal and sweetener crop in Guangxi province, China. Surveys conducted during the summer to fall seasons of 2003-2004 in northern Guangxi showed symptoms typical of a viral disease, including leaf mottling, mosaic, vein clearing, curling, and shoestring-like distortion in the field. Mechanical inoculation of sap from leaves of symptomatic plants collected from the surveyed areas caused similar symptoms on tissue culture-derived healthy Luohanguo plants. Two sequences of 0.7 and 1.6 kb with 88 and 97% identity to Papaya ringspot virus (PRSV) and Zucchini yellow mosaic virus (ZYMV) were amplified using reverse transcription-polymerase chain reaction (RT-PCR) with purified flexuous viral particles or total RNA extracted from the symptomatic Luohanguo leaves as templates with conserved degenerate potyvirus primers (1). To confirm the results, primers specific for PRSV (PP1/PP2, genome coordinates 4064-4083/5087-5069, GenBank Accession No X97251) and ZYMV (ZP1/ZP2, genome coordinates 5540-5557/7937-7920, GenBank Accession No L31350) were used to perform RT-PCR from the same RNA templates. The expected 1.0- and 2.3-kb fragments were amplified and they were 90 and 95% identical to PRSV and ZYMV in sequence, respectively. Watermelon mosaic virus was not detected. To our knowledge, this is the first report of the occurrence of PRSV and ZYMV in Luohanguo. Reference: (1) A. Gibbs et al. J. Virol. Methods 63:9, 1997.

scholarly journals Distribution of Viruses Infecting Cucurbit Crops and Isolation of Potential New Virus-Like Sequences from Weeds in Oklahoma

Plant Disease ◽  
2012 ◽  
Vol 96 (2) ◽  
pp. 243-248 ◽  
Author(s):  
Akhtar Ali ◽  
Osama Mohammad ◽  
Abeer Khattab
Keyword(s):  
Mosaic Virus ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Virus Infections ◽  
Rt Pcr ◽  
Weed Species ◽  
Field Surveys ◽  
Weed Host ◽  
Symptomatic Leaf

Field surveys were conducted from 2008 to 2010 to detect and determine the incidence of viruses in the major cucurbit-growing areas of Oklahoma. In total, 1,049 symptomatic leaf samples (890 from cucurbits, 109 from weed species, and 50 from crop plants [agricultural crops]) were collected from 90 fields in four counties (Atoka, Blaine, Jefferson, and Tulsa) of Oklahoma. Samples were tested against seven viruses, including Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus (CGMMV), Melon necrotic spot virus (MNSV), Papaya ringspot virus-watermelon strain (PRSV-W, formerly known as Watermelon mosaic virus-1), Squash mosaic virus (SqMV), Watermelon mosaic virus-2 (WMV-2), and Zucchini yellow mosaic virus (ZYMV), using dot-immunobinding assay (DIBA). Results showed the highest incidence for PRSV (51%), followed by WMV-2 (14%) and ZYMV (10%) among the collected samples. SqMV, MNSV, and CMV were detected in 3.8, 3.3, and 1.1% of the samples, respectively. None of the samples collected during surveys reacted positively against the antiserum of CGMMV. Mixed virus infections were common involving two (5.18%) or three (4.61%) viruses in various combinations. New weed host species were found to be infected with PRSV when confirmed by both DIBA and reverse-transcription polymerase chain reaction (RT-PCR). Some weed species contained possible new viruses when analyzed by random RT-PCR, followed by cloning, sequencing, and BLAST analysis with sequences in GenBank.

scholarly journals Presence and distribution of oilseed pumpkin viruses and molecular detection of Zucchini yellow mosaic virus

2009 ◽  
Vol 24 (2) ◽  
pp. 85-94 ◽  
Author(s):  
Ana Vucurovic ◽  
Aleksandra Bulajic ◽  
Ivana Djekic ◽  
Danijela Ristic ◽  
Janos Berenji ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Large Scale ◽  
Molecular Detection ◽  
Rna Extraction ◽  
Zucchini Yellow Mosaic Virus ◽  
Ringspot Virus ◽  
Yellow Mosaic Virus ◽  
Economic Losses ◽  
Watermelon Mosaic Virus ◽  
Rt Pcr

Over the past decade, intensive spread of virus infections of oilseed pumpkin has resulted in significant economic losses in pumpkin crop production, which is currently expanding in our country. In 2007 and 2008, a survey for the presence and distribution of oilseed pumpkin viruses was carried out in order to identify viruses responsible for epidemics and incidences of very destructive symptoms on cucurbit leaves and fruits. Monitoring and collecting samples of oil pumpkin, as well as other species such as winter and butternut squash and buffalo and bottle gourd with viral infection symptoms, was conducted in several localities of Vojvodina Province. The collected plant samples were tested by DAS-ELISA using polyclonal antisera specific for the detection of six most economically harmful pumpkin viruses: Cucumber mosaic virus (CMV), Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMW), Squash mosaic virus (SqMV), Papaya ringspot virus (PRSV) and Tobacco ringspot virus (TRSV) that are included in A1 quarantine list of harmful organisms in Serbia. Identification of viruses in the collected samples indicated the presence of three viruses, ZYMV, WMV and CMV, in individual and mixed infections. Frequency of the identified viruses varied depending on locality and year of investigations. In 2007, WMV was the most frequent virus (94.2%), while ZYMV was prevalent (98.04%) in 2008. High frequency of ZYMV determined in both years of investigation indicated the need for its rapid and reliable molecular detection. During this investigation, a protocol for ZYMV detection was developed and optimized using specific primers CPfwd/Cprev and commercial kits for total RNA extraction, as well as for RT-PCR. In RT-PCR reaction using these primers, a DNA fragment of approximately 1100 bp, which included coat protein gene, was amplified in the samples of infected pumkin leaves. Although serological methods are still useful for large-scale testing of a great number of samples, this protocol, due to its high sensitivity and specificity, is an important improvement in rapid diagnosis of diseases caused by this virus. In addition, the protocol provides a basis for further characterization of ZYMV isolates originating from Serbia.

scholarly journals Detection of disease in Cucurbita maxima Duch. ex Lam. caused by a mixed infection of Zucchini yellow mosaic virus, Watermelon mosaic virus, and Cucumber mosaic virus in Southeast China using a novel small RNA sequencing method

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7930
Author(s):  
Yi Wang ◽  
Pu Zhu ◽  
Qin Zhou ◽  
Xiaojun Zhou ◽  
Ziqing Guo ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Mixed Infection ◽  
Small Rnas ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Cucurbita Maxima ◽  
Rt Pcr ◽  
Virus Diseases ◽  
Southeast China

The genus Cucurbita comprises many popular vegetable and ornamental plants, including pumpkins, squashes, and gourds, that are highly valued in China as well as in many other countries. During a survey conducted in Zhejiang province, Southeast China in 2016, severe symptoms of viral infection were observed on Cucurbita maxima Duch. ex Lam. Diseased plants showed symptoms such as stunting, mosaicking, Shoe string, blistering, yellowing, leaf deformation, and fruit distortion. Approximately, 50% of Cucurbita crops produced in Jinhua were diseased, causing an estimated yield loss of 35%. In this study, we developed a method using all known virus genomes from the NCBI database as a reference to map small RNAs to develop a diagnostic tool that could be used to diagnose virus diseases of C. maxima. 25 leaf samples from different symptomatic plants and 25 leaf samples from non-symptomatic plants were collected from the experimental field of Jihua National Agricultural Technology Garden for pathogen identification. Small RNAs from each set of three symptomatic and non-symptomatic samples were extracted and sequenced by Illumina sequencing. Twenty-four different viruses were detected in total. However, the majority of the small RNAs were from Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMV), and Cucumber mosaic virus (CMV). Mixed infections of these three viruses were diagnosed in leaf samples from diseased plants and confirmed by reverse transcription PCR (RT-PCR) using primers specific to these three viruses. Crude sap extract from symptomatic leaf samples was mechanically inoculated back into healthy C. maxima plants growing under greenhouse conditions. Inoculated plants developed the same disease symptoms as those observed in the diseased plants and a mixed infection of ZYMV, WMV, and CMV was detected again by RT-PCR, thus fulfilling Koch’s postulates. The diagnostic method developed in this study involves fewer bioinformatics processes than other diagnostic methods, does not require complex settings for bioinformatics parameters, provides a high level of sensitivity to rapidly diagnose plant samples with symptoms of virus diseases and can be performed cheaply. This method therefore has the potential to be widely applied as a diagnostic tool for viruses that have genome information in the NCBI database.

scholarly journals First Report of Watermelon mosaic virus in Zucchini Squash in Bosnia and Herzegovina

Plant Disease ◽  
2014 ◽  
Vol 98 (4) ◽  
pp. 573-573 ◽  
Author(s):  
V. Trkulja ◽  
J. Stojčić ◽  
D. Kovačić ◽  
I. Stanković ◽  
A. Vučurović ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Bosnia And Herzegovina ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Post Inoculation ◽  
Rt Pcr ◽  
Sequence Comparisons ◽  
First Report ◽  
Negative Controls

Aphid-borne Watermelon mosaic virus (WMV; genus Potyvirus, family Potyviridae) is widely distributed in the Mediterranean area and is one of the most prevalent cucurbit viruses in the region (4). In July 2012, approximately 20% of zucchini squash (Cucurbita pepo L.) plants showing virus-like symptoms were observed in one field in Kukulje locality (region of Banja Luka), Bosnia and Herzegovina. Infected plants exhibited mild to severe mosaic, chlorotic mottling, and dark green vein banding, as well as puckering and leaf deformation. Symptoms mostly developed on leaves, while fruits usually only failed to develop a normal coloration. Leaves from 15 symptomatic zucchini squash plants were sampled and analyzed utilizing double-antibody sandwich (DAS)-ELISA kits (Bioreba, AG, Reinach, Switzerland) with commercial antisera specific for five commonly occurring cucurbit-infecting viruses: WMV, Zucchini yellow mosaic virus (ZYMV), Papaya ringspot virus (PRSV), Cucumber mosaic virus (CMV), and Squash mosaic virus (SqMV) (1,3,4). Commercial positive and negative controls were included in each test. WMV was detected serologically in all tested zucchini squash samples, while no presence of other tested viruses were found. Crude sap extracted from leaves of a serologically positive sample (307-12) using 0.01 M phosphate buffer (pH 7) was mechanically inoculated onto five plants of C. pepo ‘Ezra F1’ and severe mosaic accompanied by bubbling and leaf malformation was observed 14 days post-inoculation. Viral identification in all naturally and mechanically infected plants was further confirmed by conventional reverse transcription (RT)-PCR. Total RNAs were extracted with the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and RT-PCR was performed using the One-Step RT-PCR Kit (Qiagen) with specific primers WMV 5′ and WMV 3′ (4), yielding a 402- to 408-bp fragment corresponding to the N-terminal part of the coat protein (CP) gene (2). Total RNAs obtained from the Serbian WMV isolate from oil pumpkin (GenBank Accession No. JF325890) and healthy zucchini squash leaves were used as positive and negative controls, respectively. A product of the correct predicted size was obtained in all naturally and mechanically infected plants as well as positive control. No amplicon was recorded in healthy control. After purification (QIAquick PCR Purification Kit, Qiagen) the amplicon obtained from one selected isolate 307-12 was sequenced directly in both direction, aligned and compared by MEGA5 software with WMV sequences available in GenBank. Sequence comparisons revealed that the zucchini squash isolate from Bosnia and Herzegovina (KF517099) showed the highest nucleotide identity of 100% with one isolate from Serbia (FJ325891) and two Slovakian WMV isolates (GQ241712 to 13), all belonging to the classical group of WMV isolates (4). To our knowledge, this is the first report of WMV infecting zucchini squash in Bosnia and Herzegovina. Since squash and other cucurbit species represent valuable crops in Bosnia and Herzegovina, with annual production close to US$8.5 million ( http://faostat.fao.org ) and rising rapidly, the presence of a devastating pathogen like as WMV could be a serious constraint for their production. References: (1) A. Ali et al. Plant Dis. 96:243, 2012. (2) C. Desbiez et al. Arch. Virol. 152:775, 2007. (3) S. Jossey and M. Babadoost. Plant Dis. 92:61, 2008. (4) H. Lecoq and C. Desbiez. Adv. Virus Res. 84:67, 2012.

scholarly journals First Report of Tobacco ringspot virus in Pumpkin (Cucurbita pepo) in Illinois

Plant Disease ◽  
2006 ◽  
Vol 90 (10) ◽  
pp. 1361-1361 ◽  
Author(s):  
S. Jossey ◽  
M. Babadoost
Keyword(s):  
Mosaic Virus ◽  
Zucchini Yellow Mosaic Virus ◽  
The United States ◽  
Ringspot Virus ◽  
Yellow Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Watermelon Mosaic Virus ◽  
Rt Pcr ◽  
Tobacco Ringspot Virus ◽  
First Time

During a survey of commercial pumpkin and squash fields for viruses, conducted in Illinois in 2005, Tobacco ringspot virus (TRSV) was identified for the first time in symptomatic pumpkin samples collected during August and September from Douglas, Kankakee, Piatt, and Tazewell counties in one of three, one of three, one of one, and one of seven samples tested, respectively. In an earlier study from southern Illinois, the only viruses detected in pumpkins were Cucumber mosaic virus, Papaya ringspot virus, Squash mosaic virus, Watermelon mosaic virus, and Zucchini yellow mosaic virus (2). TRSV has been reported in cucurbits from some states in the United States (1). We detected TRSV in symptomatic leaves exhibiting mild mosaic with leaf yellowing using a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) kit (Agdia, Inc., Elkhart, IN). Samples were considered positive if the absorbance readings at 405 nm exceeded 3× the absorbance of the negative control. The presence of TRSV was confirmed by reverse transcription-polymerase chain reactions (RT-PCR). Total RNA was extracted from the symptomatic plants using TRIzol Reagent and reverse transcribed by M-MLV Reverse Transcriptase (Invitrogen, Carlsbad, CA). PCR was conducted using forward primer 5′-CTTGCGGCCCAAATCT ATAA-3′ and reverse primer 5′-ACTTGTGCCCAGGAGAGCTA-3′, which anneal to the conserved region in the coat protein gene. The reaction produced an amplification product of the expected size (348 bp). Hence, utilizing ELISA and RT-PCR tests, the presence of TRSV in pumpkin was determined, to our knowledge, for the first time in Illinois. References: (1) R. Provvidenti. Tobacco ringspot. Page 42 in: Compendium of Cucurbit Diseases. T. A. Zitter et al., eds. The American Phytopathological Society, St. Paul, MN. 1996. (2) S. A. Walters et al. HortScience 38:65, 2003.

scholarly journals High Prevalence of Three Potyviruses Infecting Cucurbits in Oklahoma and Phylogenetic Analysis of Cucurbit Aphid-Borne Yellows Virus Isolated from Pumpkins

Pathogens ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 53
Author(s):  
Vivek Khanal ◽  
Harrington Wells ◽  
Akhtar Ali
Keyword(s):  
Phylogenetic Analysis ◽  
Mosaic Virus ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
High Prevalence ◽  
The Us ◽  
Close Relationship ◽  
The Status ◽  
Low Incidence

Field information about viruses infecting crops is fundamental for understanding the severity of the effects they cause in plants. To determine the status of cucurbit viruses, surveys were conducted for three consecutive years (2016–2018) in different agricultural districts of Oklahoma. A total of 1331 leaf samples from >90 fields were randomly collected from both symptomatic and asymptomatic cucurbit plants across 11 counties. All samples were tested with the dot-immunobinding assay (DIBA) against the antisera of 10 known viruses. Samples infected with papaya ringspot virus (PRSV-W), watermelon mosaic virus (WMV), zucchini yellow mosaic virus (ZYMV), and cucurbit aphid-borne-yellows virus (CABYV) were also tested by RT-PCR. Of the 10 viruses, PRSV-W was the most widespread, with an overall prevalence of 59.1%, present in all 11 counties, followed by ZYMV (27.6%), in 10 counties, and WMV (20.7%), in seven counties, while the remaining viruses were present sporadically with low incidence. Approximately 42% of the infected samples were positive, with more than one virus indicating a high proportion of mixed infections. CABYV was detected for the first time in Oklahoma, and the phylogenetic analysis of the first complete genome sequence of a CABYV isolate (BL-4) from the US showed a close relationship with Asian isolates.

scholarly journals Cucurbit aphid-borne yellows virus Is Prevalent in Field-Grown Cucurbit Crops of Southeastern Spain

Plant Disease ◽  
2007 ◽  
Vol 91 (3) ◽  
pp. 232-238 ◽  
Author(s):  
M. A. Kassem ◽  
R. N. Sempere ◽  
M. Juárez ◽  
M. A. Aranda ◽  
V. Truniger
Keyword(s):  
Mosaic Virus ◽  
Zucchini Yellow Mosaic Virus ◽  
Rflp Analysis ◽  
Yellow Mosaic Virus ◽  
Multiple Infections ◽  
Watermelon Mosaic Virus ◽  
Limited Information ◽  
Growing Seasons ◽  
Beet Pseudo Yellows Virus ◽  
The Impact

Despite the importance of field-grown cucurbits in Spain, only limited information is available about the impact of disease on their production. During the 2003 and 2004 growing seasons, systematic surveys were carried out in open field melon (Cucumis melo) and squash (Cucurbita pepo) crops of Murcia Province (Spain). The fields were chosen with no previous information regarding their sanitation status, and samples were taken from plants showing viruslike symptoms. Samples were analyzed using molecular hybridization to detect Beet pseudo-yellows virus (BPYV), Cucurbit aphid-borne yellows virus (CABYV), Cucumber mosaic virus (CMV), Cucumber vein yellowing virus (CVYV), Cucurbit yellow stunting disorder virus (CYSDV), Melon necrotic spot virus (MNSV), Papaya ringspot virus (PRSV), Watermelon mosaic virus (WMV), and Zucchini yellow mosaic virus (ZYMV). We collected 924 samples from 48 field plots. Out of these, almost 90% were infected by at least one of the viruses considered, usually CABYV, which was present in 83 and 66% of the melon and squash samples, respectively. In the case of melon, CYSDV, BPYV, and WMV followed CABYV in relative importance, with frequencies of around 20 to 30%, while in squash, CVYV and BPYY showed frequencies between 28 and 21%. The number of multiple infections was very high, 66 and 56% of the infected samples of melon and squash, respectively, being afflicted. CABYV was present in all multiple infections. The high incidence of CABYV in single and multiple infections suggests that this virus may well become an important threat for cucurbit crops in the region. Restriction fragment length polymorphism (RFLP) analysis revealed that CABYV isolates can be grouped into two genetic types, both of which seemed to be present during the 2003 epidemic episode, but only one of the types was found in 2004.

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