scholarly journals Genetic Diversity of Potyviruses Associated with Tulip Breaking Syndrome

Plants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1807
Author(s):  
János Ágoston ◽  
Asztéria Almási ◽  
Katalin Salánki ◽  
László Palkovics

Tulip breaking is economically the most important viral disease of modern-day tulip growing. It is characterized by irregular flame and feather-like patterns in the flowers and mosaic on the foliage. Thirty-two leaf samples were collected from cultivated tulip plants showing tulip breaking syndrome from Hungary in 2017 and 2018. Virus identification was performed by serological (ELISA) and molecular (RT-PCR) methods. All samples proved to be infected with a potyvirus and evidence was provided that three potyvirus species could be identified in the samples: Lily mottle virus (LMoV), Tulip breaking virus (TBV) and Rembrandt tulip-breaking virus (ReTBV). Recombination prediction accomplished with Recombination Detection Program (RDP) v4.98 revealed potential intraspecies recombination in the case of TBV and LMoV. Phylogenetic analyses of the coat protein (CP) regions proved the monophyletic origin of these viruses and verified them as three different species according to current International Committee on Taxonomy of Viruses (ICTV) species demarcation criteria. Based on these results, we analyzed taxonomic relations concerning potyviruses associated with tulip breaking syndrome. We propose the elevation of ReTBV to species level, and emergence of two new subgroups in ReTBV.

2019 ◽  
Vol 31 (2) ◽  
pp. 263-268
Author(s):  
Ilyas Deligoz ◽  
Mehmet Ali Sevik

AbstractThe tulip (Tulipa sp.) is one of the most important ornamental bulbous plants, which has been cultivated as a cut-flower, potted, and garden plant, and used for landscaping in Turkey. This study investigated the occurrence of a viral disease in the tulip cultivars Strong Gold, Pretty Woman and Purple Prince that causes striping of the leaves, flames of different colours on the petals and mosaic patterns on the leaves, in Samsun province of Turkey. Surveys of virus-infected tulip plants were carried out in the Middle Black Sea Region of Turkey in 2015-2016. A total of 212 samples were collected from four locations and checked by biological, serological and molecular methods for the presence of the Tulip breaking virus (TBV). TBV was detected in the leaves and flowers by double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) in the tulip cultivars (15.5%) tested from Samsun province. TBV infection was found at the highest rate in the cultivar Strong Gold (19.7%), followed by Pretty Woman (14.1%) and Purple Prince (12.8%). The presence of TBV in samples was further confirmed by reverse transcription polymerase chain reaction (RT-PCR) assays. This is the first report on TBV naturally infecting tulips in Samsun province, Turkey.


Plant Disease ◽  
2013 ◽  
Vol 97 (11) ◽  
pp. 1517-1517 ◽  
Author(s):  
M. A. V. Alexandre ◽  
L. M. L. Duarte ◽  
E. B. Rivas ◽  
E. W. Kitajima ◽  
R. Harakava

Zamioculcas zamiifolia (Lodd.) Engl. (“Zanzibar Gem,” “ZZ plant”) is the monotypic species of the genus belonging to the family Araceae. It is a stemless perennial plant native to Africa, from Kenya to South Africa, that produces succulent rhizomes at the base of its attractive dark green and glossy foliage. Symptoms of mosaic and foliar distortion were observed on a plant purchased at an ornamental plants shop in São Paulo state, Brazil. In order to identify the causal agent, transmission and serological tests, as well as electron microscopy (EM) observations, reverse transcription (RT)-PCR, and sequencing were carried out. EM observations revealed the presence of elongated, flexuous viral particles in foliar extracts and cytoplasmic lamellar aggregates of type II lamellar inclusions (Edwardson's classification), in thin sections. No symptoms were induced following mechanical inoculation on Chenopodium amaranticolor, C. murale, Gomphrena globosa, Nicotiana megalosiphon, N. debneyii, nor on the aroids Philodendron scandens, P. selloum, Dieffenbachia amoena, Colocasia esculenta, and Z. zamiifolia. Up to 2 months after inoculation, plants were still symptomless, and the virus was not detected by RT-PCR. The indirect ELISA tests were negative with antisera against Dasheen mosaic virus (gift from F. W. Zettler, University of Florida) and Turnip mosaic virus (gift from P. Roggero, IFA, Turin, Italy). RT-PCR performed on the original purchased ornamental plant with potyvirus-specific primers (CI-R = ACICCRTTYTCDATDATRTTIGTIGC and CI-F = GGIVVIGTIGGIWSIGGIAARTCIAC) targeting the cytoplasmic inclusion protein cistron of the potyvirus genome produced a fragment of approximately 650 bp (GenBank Accession No. KC990386). The sequence was similar to those of potyvirus species with nucleotide identity, determined by PAUP v.4.0b10 for Macintosh, ranging from 64% for Pokeweed mosaic virus (JQ609065) to 93% for Konjac mosaic virus KoMV-F (NC007913). KoMV has been detected in aroid species in Taiwan, India, Korea, Japan (1,2), Germany, and The Netherlands (3,4). This is the first report of a viral disease on Z. zamiifolia and of KoMV in the Americas. Such information along with the vegetative propagation of ZZ plants strongly suggests that KoMV is spread worldwide. References: (1) P. Manikonda et al. J. Phytopathol. 159:133, 2011. (2) M. Nishiguchi et al. Arch Virol. 151:1643, 2006. (3) D.-E. Lesemann and S. Winter. Acta Hort. 568:135, 2002. (4) K. Pham et al. Acta Hort. 568:143, 2002.


Plant Disease ◽  
2003 ◽  
Vol 87 (4) ◽  
pp. 446-446 ◽  
Author(s):  
N. L. Robertson

The Matanuska-Susitna Valley is one of the most fertile regions in Alaska for growing cool-season vegetables. Barley (Hordeum vulgare) and oat (Avena sativa) crops are also sown for animal feed and green manure. The most damaging and widely distributed viral disease of small grains worldwide is barley yellow dwarf (BYD), caused by several species from two genera in the family Luteoviridae: luteovirus (Barley yellow dwarf virus [BYDV-MAV and BYDV-PAV]) and polerovirus (Cereal yellow dwarf virus [CYDV-RPV, formerly BYDV-RPV]) and three unassigned species (BYDV-RMV, BYDV-SGV, and BYDV-GPV) (2,4). Even though barley and oat have been grown in Alaska for more than 50 years, BYD has not been documented in small grains in this region. During September 2001, barley plants with bright yellow leaves were collected from five barley fields near Palmer. Three plants from each field were assayed using a reverse transcription-polymerase chain reaction (RT-PCR) protocol targeting members of the luteoviridae (3). The resulting ≈530-bp PCR product and its restriction fragment length polymorphism (RFLP) produced by digestion with NdeII implied that plants were infected with BYDV-PAV. In September 2002, three of the five sites were surveyed again for BYDV. Two of the fields (BF-1 and BF-2) had been replanted with barley and the other (OF-3) was planted with oats. Leaf samples from 36 symptomatic barley plants from each field and 60 symptomatic oat plants were randomly collected and stored at -80°C. In 2002, in addition to RT-PCR and RFLP analyses, enzyme-linked immunosorbent assays (ELISA) using Agdia kits (Agdia, Elkhart, IN) for BYDV-PAV, CYDV-RPV, and BYDV-SGV were also performed (1). First, RT-PCR and RFLP were completed on all samples using 0.5 g of tissue. Of samples from BF-1, BF-2, and OF-3, 61, 100, and 70%, respectively, generated luteoviridae-specific fragments. The RFLP profiles from barley were all PAV-like, whereas 71% of oat samples were PAV-like, and 29% were of an unknown pattern. No bands were observed from apparently healthy field plants. ELISA (0.2 g of tissue) was performed on all PCR-positive samples, resulting in 22, 97, and 33% detection for BYDV-PAV from BF-1, BF-2, and OF-3, respectively. An additional 29% of oat samples (OF-3) tested positive for CYDV-RPV, whereas none of the barley plants tested positive. One oat plant had a mixed infection with both PAV and RPV profiles, and all oat plants with the unidentified RFLP pattern were serologically positive for RPV. No BYDV-SGV was detected in either barley or oats. The PCR assay was clearly more sensitive than ELISA, especially for plants that had mature and necrotic tissue, which were predominately found in BF-1 and OF-3. Based on these direct tests on the coat protein's nucleic acid (PCR) and serology (ELISA), it is concluded that two distinct viruses, BYDV-PAV and CYDV-RPV, were found in oats, whereas only the former was found in barley. To my knowledge, this is the first report of luteovirus and polerovirus infection in small grains in Alaska. References: (1) M. F. Clark and A. N. Adams. J. Gen. Virol. 34, 475, 1977. (2) C. J. D'Arcy and P. A. Burnett. Barley Yellow Dwarf: 40 Years of Progress. The American Phytopathological Society, St. Paul, MN, 1995. (3) N. L. Robertson and R. French. J. Gen. Virol. 72,1473, 1991. (4) M. H. V. van Regenamortel et al. Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses. Academic Press, NY, 2000.


Viruses ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 66
Author(s):  
Zoltán László ◽  
Péter Pankovics ◽  
Gábor Reuter ◽  
Attila Cságola ◽  
Ádám Bálint ◽  
...  

Most picornaviruses of the family Picornaviridae are relatively well known, but there are certain “neglected” genera like Bopivirus, containing a single uncharacterised sequence (bopivirus A1, KM589358) with very limited background information. In this study, three novel picornaviruses provisionally called ovipi-, gopi- and bopivirus/Hun (MW298057-MW298059) from enteric samples of asymptomatic ovine, caprine and bovine respectively, were determined using RT-PCR and dye-terminator sequencing techniques. These monophyletic viruses share the same type II-like IRES, NPGP-type 2A, similar genome layout (4-3-4) and cre-localisations. Culture attempts of the study viruses, using six different cell lines, yielded no evidence of viral growth in vitro. Genomic and phylogenetic analyses show that bopivirus/Hun of bovine belongs to the species Bopivirus A, while the closely related ovine-origin ovipi- and caprine-origin gopivirus could belong to a novel species “Bopivirus B” in the genus Bopivirus. Epidemiological investigation of N = 269 faecal samples of livestock (ovine, caprine, bovine, swine and rabbit) from different farms in Hungary showed that bopiviruses were most prevalent among <12-month-old ovine, caprine and bovine, but undetectable in swine and rabbit. VP1 capsid-based phylogenetic analyses revealed the presence of multiple lineages/genotypes, including closely related ovine/caprine strains, suggesting the possibility of ovine–caprine interspecies transmission of certain bopiviruses.


Animals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 277
Author(s):  
Eleonora Chelli ◽  
Elisabetta Suffredini ◽  
Paola De Santis ◽  
Dario De Medici ◽  
Santina Di Bella ◽  
...  

In Europe, foodborne transmission has been clearly associated to sporadic cases and small clusters of hepatitis E in humans linked to the consumption of contaminated pig liver sausages, raw venison, or undercooked wild boar meat. In Europe, zoonotic HEV-genotype 3 strains are widespread in pig farms but little information is available on the prevalence of HEV positive pigs at slaughterhouse. In the present study, the prevalence of HEV-RNA positive pigs was assessed on 585 animals from 4 abattoirs located across Italy. Twenty-one pigs (3.6%) tested positive for HEV in either feces or liver by real-time RT-PCR. In these 21 pigs, eight diaphragm muscles resulted positive for HEV-RNA. Among animals collected in one abattoir, 4 out of 91 plasma tested positive for HEV-RNA. ELISA tests for the detection of total antibodies against HEV showed a high seroprevalence (76.8%), confirming the frequent exposure of pigs to the virus. The phylogenetic analyses conducted on sequences of both ORF1 and ORF2 fragments, shows the circulation of HEV-3c and of a novel unclassified subtype. This study provides information on HEV occurrence in pigs at the slaughterhouse, confirming that muscles are rarely contaminated by HEV-RNA compared to liver, which is the most frequently positive for HEV.


Plant Disease ◽  
2017 ◽  
Vol 101 (5) ◽  
pp. 726-733 ◽  
Author(s):  
Mengpei Guo ◽  
Yinbing Bian ◽  
Jinjie Wang ◽  
Gangzheng Wang ◽  
Xiaolong Ma ◽  
...  

A new partitivirus named Lentinula edodes partitivirus 1 (LePV1) was isolated from a diseased L. edodes strain with severe degeneration of the mycelium and imperfect browning in bag cultures. The nucleotide sequences of LePV1 dsRNA-1 and dsRNA-2 were determined; they were 2,382 bp and 2,245 bp in length, and each contained a single ORF encoding RNA-dependent RNA polymerase (RdRp) and coat protein (CP), respectively. The purified virus preparation contained isometric particles 34 nm in diameter encapsidating these dsRNAs. Phylogenetic analyses showed LePV1 to be a new member of Betapartitivirus, with the RdRp sequence most closely related to Grapevine partitivirus. RT-PCR analysis showed that 27 of the 56 Chinese L. edodes core collection strains carry LePV1, with the virus being more common in wild strains than cultivated strains. In addition, qPCR analysis suggested that coinfection with L. edodes mycovirus HKB (LeV-HKB) could increase replication of the RdRp gene of LePV1. This study may be essential for the development of more accurate disease diagnostics and the formulation of control strategies for viral diseases in L. edodes.


2012 ◽  
Vol 2012 ◽  
pp. 1-12 ◽  
Author(s):  
Denise Gonçalves ◽  
Rafael de Queiroz Prado ◽  
Eric Almeida Xavier ◽  
Natália Cristina de Oliveira ◽  
Paulo Marcos da Matta Guedes ◽  
...  

Dengue fever is a noncontagious infectious disease caused by dengue virus (DENV). DENV belongs to the familyFlaviviridae, genusFlavivirus, and is classified into four antigenically distinct serotypes: DENV-1, DENV-2, DENV-3, and DENV-4. The number of nations and people affected has increased steadily and today is considered the most widely spread arbovirus (arthropod-borne viral disease) in the world. The absence of an appropriate animal model for studying the disease has hindered the understanding of dengue pathogenesis. In our study, we have found that immunocompetent C57BL/6 mice infected intraperitoneally with DENV-1 presented some signs of dengue disease such as thrombocytopenia, spleen hemorrhage, liver damage, and increase in production of IFNγand TNFαcytokines. Moreover, the animals became viremic and the virus was detected in several organs by real-time RT-PCR. Thus, this animal model could be used to study mechanism of dengue virus infection, to test antiviral drugs, as well as to evaluate candidate vaccines.


2015 ◽  
Vol 35 (6) ◽  
pp. 557-561 ◽  
Author(s):  
Juliana F. Cargnelutti ◽  
Eduardo K. Masuda ◽  
Mariana G. Neuls ◽  
Rudi Weiblen ◽  
Eduardo F. Flores

Abstract: Canid herpesvirus 1 (CHV-1) is a widespread pathogen of dogs and produces infertility, abortions and severe systemic disease in young puppies. Clinical data indicate the circulation of CHV-1 among Brazilian dogs yet definitive diagnosis has rarely been accomplished. This article describes the clinicopathological findings of four independent cases/outbreaks of neonatal disease by CHV-1 in Bulldog puppies followed by virus identification and genetic characterization. Three events occurred in a kennel holding dogs of different breeds at reproductive age (March 2013, October 2013 and April 2014). Puppies from three French or English Bulldog litters, aging 9 to 30 days were affected, presenting dyspnea, agonic breathing, pale mucous, abdominal pain and tension, evolving to death within about 24 hours. At necropsy, the puppies presented necrohemorrhagic hepatitis, multifocal and moderate necrohemorrhagic nephritis and fibrinonecrotic interstitial pneumonia. Virus isolation was positive in clinical specimens from one litter and CHV-1 DNA was detected by PCR in tissues from all four cases. Virus-neutralizing assays with samples of the affected kennel revealed 9/12 adult animals with high antibody titers to CHV-1. Nucleotide sequencing of glycoprotein B, C and D genes revealed 99-100% of identity among the viruses and with CHV-1 sequences available in GenBank. Phylogenetic analyses of gC sequences showed a segregation of the samples, even among three isolates from the same kennel. These findings support CHV-1 infection as the cause of disease and death in these dog litters, reinforcing the need for correct etiologic diagnosis, prevention and immunization against CHV-1 in dogs from Southern Brazil.


2021 ◽  
Author(s):  
Michela Bulfoni ◽  
Emanuela Sozio ◽  
Barbara Marcon ◽  
Maria De Martino ◽  
Daniela Cesselli ◽  
...  

Background: Since the beginning of the pandemic, clinicians and researchers have been searching for alternative tests to improve screening and diagnosis of SARS-CoV-2 infection. Currently, the gold standard for virus identification is the nasopharyngeal (NP) swab. Saliva samples, however, offer clear practical and logistical advantages but due to lack of collection, transport, and storage solutions, high-throughput saliva-based laboratory tests are difficult to scale up as a screening or diagnostic tool. With this study, we aimed to validate an intra-laboratory molecular detection method for SARS-CoV-2 on saliva samples collected in a new storage and inactivating solution, comparing the results to NP swabs to determine the difference in sensitivity between the two tests. Methods: In this study, 156 patients (cases) and 1005 asymptomatic subjects (controls) were enrolled and tested simultaneously for the detection of the SARS-CoV-2 viral genome by RT-PCR on both NP swab and saliva samples. Saliva samples were collected in a preservative and inhibiting saline solution (Biofarma Srl). Internal method validation was performed to standardize the entire workflow for saliva samples. Results: The identification of SARS-CoV-2 conducted on saliva samples showed a clinical sensitivity of 95.1% and specificity of 97.8% compared to NP swabs. The positive predictive value (PPV) was 81% while the negative predictive value (NPV) was 99.5%. Test concordance was 97.6% (Cohen's Kappa=0.86; 95% CI 0.81-0.91). The LoD of the test was 5 viral copies for both samples. Conclusions: RT-PCR assays conducted on a stored saliva sample achieved similar performance to those on NP swabs and this may provide a very effective tool for population screening and diagnosis. Collection of saliva in a stabilizing solution makes the test more convenient and widely available; furthermore, the denaturation properties of the solution reduces the infective risks belonging to sample manipulation.


Plant Disease ◽  
2010 ◽  
Vol 94 (2) ◽  
pp. 276-276 ◽  
Author(s):  
W. Menzel ◽  
S. Winter ◽  
K. R. Richert-Pöggeler

Hollyhocks are popular garden plants and selected cultivars of Alcea rosea (family Malvaceae) are widespread in Germany. In spring 2009, dozens of A. rosea plants displaying strong vein clearing and veinal yellowing symptoms were found in private gardens in Hannover, Lower Saxony. Electron microscopic examinations of negatively stained adsorption preparations of five randomly selected samples of symptomatic plants or their offshoots revealed flexuous filamentous particles resembling those of potyviruses. Sap extracts also reacted strongly positive in an antigen coated plate (ACP)-ELISA with the broad-spectrum potyvirus antiserum AS-0573/I (DSMZ, Braunschweig, Germany). RNA extracts (RNeasy Kit, Qiagen, Valencia, CA) of the above mentioned leaf samples were used as templates in reverse transcription (RT)-PCR assays with potyvirus specific primers (2) that have been shown to amplify the 3′ terminus of the genome of many potyvirus species. For extracts from symptomatic samples, this resulted in a consistent amplification of an ~1.6-kbp fragment, whereas no products were obtained from RNA extracts of asymptomatic plants. From one positive sample, the amplified fragment was cloned and one clone was partially sequenced. The nucleotide (nt) and amino acid sequences showed the highest identities (81 to 83% and 87 to 90%, respectively) to GenBank sequences FJ539084, FM212972, EU884405, and FJ561293 of the potyvirus Malva vein clearing virus (MVCM). On the basis of these identity values and according to the species demarcation criteria in the genus Potyvirus, the virus can be regarded as a German isolate of the recently sequenced MVCV (3,4). Direct sequencing of the 5′-end of the amplified RT-PCR fragment revealed sequences of only one potyvirus species. The virus isolate has been submitted to the DSMZ Plant Virus Collection (Braunschweig, Germany) under accession PV-0963 and the sequence obtained from the cloned cDNA is deposited in GenBank (GQ856544). In addition, sap from affected leaves was mechanically inoculated onto sets of herbaceous indicator plants (Chenopodium quinoa, C. foliosum, C. murale, C. amaranticolor, Datura stramonium, Nicotiana benthamiana, N. hesperis, Petunia hybrida, and Solanum lycopersicum) of which only C. quinoa plants became infected. Symptoms of weak chlorosis along and beside veins of inoculated leaves, but not systemic leaves, became visible 2 weeks postinoculation. Symptomatic leaves contained flexuous filamentous particles and ACP-ELISA and RT-PCR confirmed virus presence. The partially sequenced amplicon showed 99% nt identity to the sequence from the cloned cDNA. To our knowledge, this is the first report of a MVCV isolate naturally occurring in A. rosea and C. quinoa is the first host identified that does not belong to the plant family Malvaceae. In contrast, the MVCV isolate used in the host range study of Lunello et al. (4) did not infect A. rosea and C. quinoa, confirming previous host range descriptions by Brunt et al. (1). Since MVCV infections of hollyhocks seem to cause only leaf symptoms and do not noticeably affect growth or flowering of the plants, this will hopefully not impair the usability of this popular garden plant. References: (1) A. A. Brunt et al. Descriptions and Lists from the VIDE Database. Online publication. Version: 16th January, 1997. (2) J. Chen et al. Arch. Virol. 146:757, 2001. (3) A. Hein Phytopathol. Z. 28:205, 1957. (4) P. Lunello et al. Virus Res. 140:91, 2009.


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