scholarly journals Effect of Ozone on Inactivation of Purified Pepper Mild Mottle Virus and Contaminated Pepper Seed

2021 ◽  
pp. PHYTOFR-09-20-0
Author(s):  
John R. Stommel ◽  
Judith M. Dumm ◽  
John Hammond

Pepper mild mottle virus (PMMoV) is a major viral pathogen of pepper (Capsicum spp.). PMMoV is readily mechanically transmitted and is seed transmissible. Trisodium phosphate (TSP) treatment is commonly used to reduce the level of viable PMMoV in contaminated seed. Ozone is efficacious in disinfecting fungal-contaminated seed and disrupting bacterial and viral pathogens on various substrates. The purpose of this study was to evaluate efficacy of ozone and chemical treatment on PMMoV viability. Treated pepper seed infectivity was evaluated via bioassay with Nicotiana benthamiana. Symptoms of PMMoV infection were not observed in bioassays of TSP-treated seed. Sufficient viable PMMoV remained on ozone-treated seed to cause infection, which was confirmed by ELISA. Neither treatment affected seed germination. Ozone treatment of purified PMMoV was assessed to determine the extent, if any, of PMMoV inactivation by ozone. At the low PMMoV concentration (0.01 mg/ml), 14-h ozone exposure eliminated infectivity as determined by N. benthamiana bioassays with ELISA confirmations. At the higher PMMoV concentration (0.1 mg/ml), ozone treatment was insufficient to prevent infection. Ozone inactivation of purified PMMoV was quantified via bioassay using the local lesion host N. glutinosa and quantitative real-time PCR. Ozone exposure reduced lesion counts and PMMoV concentration, and PMMoV degradation increased with exposure time. Although PMMoV infection was eliminated at the low PMMoV concentration, bioassays using naturally infected seed and purified PMMoV preparations at relatively higher concentrations demonstrated that ozone is not efficacious as a standard treatment to sufficiently reduce levels of infective PMMoV in contaminated pepper seed. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

1998 ◽  
Vol 11 (12) ◽  
pp. 1253-1257 ◽  
Author(s):  
P. Gilardi ◽  
I. García-Luque ◽  
M. T. Serra

The pepper mild mottle virus (PMMoV-S) (an L3 hypersensitive response [HR]-inducer strain) coat protein was expressed in Capsicum chinense (L3L3) plants with the heterologous potato virus X (PVX)-based expression system. The chimeric virus was localized in the inoculated leaves and induced the HR, thus indicating that the tobamoviral sequences that affect induction of the HR conferred by the L3 resistance gene reside in the coat protein gene. Furthermore, transient expression of the PMMoV-S coat protein in C. chinense leaves by biolistic co-bombardment with a plasmid expressing the β-glucuronidase (GUS) gene leads to the induction of cell death and expression of host defense genes. Thus, the coat protein of PMMoV-S is the elicitor of the Capsicum spp. L3 resistance gene-mediated HR.


Author(s):  
Brienna L. Anderson-Coughlin ◽  
Shani Craighead ◽  
Alyssa Kelly ◽  
Samantha Gartley ◽  
Adam Vanore ◽  
...  

Enteric viruses (EV) are the largest contributors to foodborne illness and outbreaks globally. Their ability to persist in the environment, coupled with the challenges experienced in environmental monitoring create a critical aperture through which agricultural crops may become contaminated. This study involved a seventeen-month investigation of select human enteric viruses and viral indicators in non-traditional irrigation water sources, surface and reclaimed waters, in the Mid-Atlantic region of the United States. Real-time quantitative PCR was used for detection of Aichi virus, hepatitis A virus, noroviruses GI and GII. Pepper mild mottle virus (PMMoV), a common viral indicator of human fecal contamination, was also evaluated along with atmospheric (air and water temperature, cloud cover, and precipitation 24 h, 7 d and 14 d prior to sample collection) and physicochemical (dissolved oxygen, pH, salinity and turbidity) data to determine if there were any association between EV and measured parameters. EV were detected more frequently in reclaimed waters (32%, n=22) than in surface waters (4%, n=49) similar to PMMoV detection frequency in surface (33%, n=42) and reclaimed (67%, n=21) water. Our data show a significant correlation between EV and PMMoV (R2=0.628; p<0.05) detection in reclaimed water samples, but not in surface waters (R2=0.476; p=0.78). Water salinity significantly affected the detection of both EV and PMMoV (p<0.05) as demonstrated by logistic regression analyses. These results provide relevant insights into the extent and degree of association between human enteric (pathogenic) viruses and water quality data in Mid-Atlantic surface and reclaimed waters as potential sources for agricultural irrigation. IMPORTANCE Microbiological analysis of agricultural waters is fundamental to ensure microbial food safety. The highly variable nature of non-traditional sources of irrigation water are particularly difficult to test for the presence of viruses. Multiple characteristics influence viral persistence in a water source as well as affect the recovery and detection methods which are employed. Testing for a suite of viruses in water samples is often too costly and labor intensive, making identification of suitable indicators for viral pathogen contamination necessary. The results from this study address two critical data gaps: enteric virus prevalence in surface and reclaimed waters of the Mid-Atlantic region of the United States and subsequent evaluation of physicochemical and atmospheric parameters used to inform the potential for use of indicators of viral contamination.


Author(s):  
Jefferson B. Vélez-Olmedo ◽  
Cesar E. Fribourg ◽  
Fernando L. Melo ◽  
Tatsuya Nagata ◽  
Athos S. de Oliveira ◽  
...  

Tobamoviruses are often referred to as the most notorious viral pathogens of pepper crops. These viruses are not transmitted by invertebrate vectors, but rather by physical contact and seeds. In this study, pepper plants displaying mild mottle and mosaic symptoms were sampled in four different regions of Peru. Upon double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) tests, seven samples cross-reacted weakly with antibodies against pepper mild mottle virus (PMMoV), suggesting the presence of tobamoviruses. When employing RT-PCR, conserved primers amplified cDNA fragments of viruses from two putative new tobamovirus species in the samples. The complete genome of two representative isolates were, therefore, sequenced and analysed in silico. These viruses, which were tentatively named yellow pepper mild mottle virus (YPMMoV) and chilli pepper mild mottle virus (CPMMoV), shared highest nucleotide genome sequence identities of 83 and 85 % with bell pepper mottle virus (BpeMV), respectively. Mechanical inoculation of indicator plants with YPMMoV and CPMMoV isolates did not show any obvious differences in host ranges. These viruses were also inoculated mechanically on pepper plants harbouring different resistance L alleles to determine their pathotypes. Pepper plants carrying unfunctional L alleles (L 0) to tobamoviruses were infected by all isolates and presented differential symptomatology for YPMMoV and CPMMoV. On the other hand, pepper plants carrying L 1, L 2, L 3 and L 4 alleles were resistant to all isolates, indicating that these viruses belong to pathotype P0.


2004 ◽  
Vol 29 (6) ◽  
pp. 670-675 ◽  
Author(s):  
Marcelo Eiras ◽  
Alexandre L. R. Chaves ◽  
Silvia R. Moreira ◽  
Jansen de Araujo ◽  
Addolorata Colariccio

Sementes de pimenta (Capsicum baccatum) 'Dedo de Moça' destinadas ao plantio comercial e adquiridas no município de São Paulo, SP, analisadas quanto à presença de vírus, por meio de testes biológicos e sorológicos revelaram-se infetadas por uma estirpe do Pepper mild mottle virus (PMMoV). Para confirmar a identidade do isolado, promoveu-se a RT-PCR com oligonucleotídeos que flanqueiam a ORF da capa protéica de espécies do gênero Tobamovirus do subgrupo 1. Os fragmentos de DNA amplificados, quando seqüenciados e comparados com outros isolados de tobamovírus depositados no GenBank, apresentaram valores de identidade de nucleotídeos entre 94 e 100% com outras seqüências de PMMoV, inferiores a 75% para as demais espécies de tobamovírus do subgrupo I (Tobacco mosaic virus, Tomato mosaic virus e Odontoglossum ringspot virus) e 65% para os tobamovírus dos subgrupos II e III. O PMMoV-BR revelou 100% de identidade com isolados japoneses, sugerindo que este patógeno pode ter sido introduzido daquele país. A seqüência de aminoácidos deduzidos da capa protéica indicou também, que este isolado não é capaz de quebrar a resistência do gene L3 de Capsicum spp. Fato confirmado pelos sintomas causados nas hospedeiras diferenciais de Capsicum spp., verificando-se que este isolado não foi capaz de infetar plantas de C. chinense (L3) e C. chacoense (L4). Estes resultados confirmaram a importância da caracterização dos isolados de tobamovírus, fundamental para adequação de medidas de controle, principalmente, prevenindo a entrada e posterior disseminação do patógeno em novas áreas de cultivo.


2009 ◽  
Vol 35 (1) ◽  
pp. 39-43 ◽  
Author(s):  
Márcia Aparecida Cezar ◽  
Renate Krause-Sakate ◽  
Marcelo Agenor Pavan ◽  
Cyro Paulino da Costa

A resistência em Capsicum spp a tobamovírus é governada pelos genes L¹ a L4. Baseado na capacidade de alguns isolados suplantarem a resistência destes genes, os tobamovírus podem ser classificados nos patótipos P0, P1, P1-2 e P1-2-3. No Brasil, até o momento as três espécies de tobamovírus conhecidas são: Tobacco mosaic virus (TMV), Tomato mosaic virus (ToMV), pertencentes aos patótipos P0 e Pepper mild mottle virus (PMMoV) pertencente ao patótipo P1-2, respectivamente e podem infectar pimentas e pimentões. Oitenta e seis genótipos de pimentão e pimenta foram avaliados quanto à resistência a tobamovírus, sendo 62 de Capsicum annuum, 18 de C. baccatum e seis de C. chinense. Oito acessos de C. annuum, seis de C. baccatum e os acessos ICA #39, Pimenta de cheiro e PI 152225 de C. chinense apresentaram reação de hipersensibilidade ao ToMV, enquanto que o acesso Ancho de C. annuum foi considerado tolerante, permanecendo assintomático, porém permitindo a recuperação do vírus quando inoculado em Nicotiana glutinosa. Para o PMMoV patótipo P1,2 foram avaliados os acessos de pimentão e pimenta considerados resistentes ao ToMV. Somente o PI 152225 de C. chinense desencadeou reação de hipersensibilidade ao PMMoV, sendo fonte potencial de resistência para programas de melhoramento a este vírus no Brasil.


2007 ◽  
Vol 97 (7) ◽  
pp. 787-793 ◽  
Author(s):  
Yoshikatsu Genda ◽  
Ayami Kanda ◽  
Hiroyuki Hamada ◽  
Kyoko Sato ◽  
Jun Ohnishi ◽  
...  

The Capsicum spp. L genes (L1 to L4) confer resistance to tobamoviruses. Currently, the L4 gene from Capsicum chacoense is the most effective resistance gene and has been used widely in breeding programs in Japan which have developed new resistant cultivars against Pepper mild mottle virus (PMMoV). However, in 2004, mild mosaic symptoms began appearing on the leaves of commercial pepper plants in the field which possessed the L4 resistance gene. Serological and biological assays on Capsicum spp. identified the causal virus strain as a previously unreported pathotype, P1,2,3,4. PMMoV sequence analysis of the virus and site-directed mutagenesis using a PMMoV-J of the P1,2 pathotype revealed that two amino acid substitutions in the coat protein, Gln to Arg at position 46 and Gly to Lys at position 85, were responsible for overcoming the L4 resistance gene.


1997 ◽  
Vol 10 (1) ◽  
pp. 107-113 ◽  
Author(s):  
A. de la Cruz ◽  
L. López ◽  
F. Tenllado ◽  
J. R. Díaz-Ruíz ◽  
A. I. Sanz ◽  
...  

In Capsicum, the resistance against tobamoviruses conferred by the L2 gene is effective against all but one of the known tobamoviruses. Pepper mild mottle virus (PMMoV) is the only virus which escapes its action. To identify the viral factors affecting induction of the hypersensitive reaction (HR) mediated by the Capsicum spp. L2 resistance gene, we have constructed chimeric viral genomes between paprika mild mottle virus (PaMMV) (a virus able to induce the HR) and PMMoV. A hybrid virus with the PaMMV coat protein gene substituted in the PMMoV-S sequences was able to elicit the HR in Capsicum frutescens (L2L2) plants. These data indicate that the sequences that affect induction of the HR mediated by the L2 resistance gene reside in the coat protein gene. Furthermore, a mutant that codes for a truncated coat protein was able to systemically spread in these plants. Thus, the elicitation of the host response requires the coat protein and not the RNA.


1989 ◽  
Vol 63 (5) ◽  
pp. 2198-2203 ◽  
Author(s):  
E Rodríguez-Cerezo ◽  
A Moya ◽  
F García-Arenal

2021 ◽  
Author(s):  
Agnes S Montgomery ◽  
Michael B Lustik ◽  
Susan A Reichert-Scrivner ◽  
Ronald L Woodbury ◽  
Milissa U Jones ◽  
...  

ABSTRACT Introduction Acute respiratory diseases account for a substantial number of outpatient visits and hospitalizations among U.S. military personnel, significantly affecting mission readiness and military operations. We conducted a retrospective analysis of respiratory viral pathogen (RVP) samples collected from U.S. military personnel stationed in Hawaii and tested at Tripler Army Medical Center from January 2014 to May 2019 in order to describe the etiology, distribution, and seasonality of RVP exposure in a military population. Materials and Methods Samples were analyzed by viral culture or multiplex PCR. Distribution of respiratory viruses over time was analyzed as well as subject demographic and encounter data. Presenting signs and symptoms were evaluated with each RVP. Results A total of 2,576 military personnel were tested, of which 726 (28.2%) were positive for one or more RVP. Among positive tests, the three most common viral pathogens detected were influenza A (43.0%), rhinovirus (24.5%), and parainfluenza (7.6%). Symptoms were generally mild and most frequently included cough, fever, and body aches. Conclusion Our study evaluated respiratory virus prevalence, seasonality, and association with clinical symptoms for military personnel in an urban tropical setting in Oahu, HI, over a 5-year period. We show that viral prevalence and seasonality in Hawaii are distinct from those of the CONUS. Results contribute to the broader understanding of seasonality, clinical manifestation, and demographics of RVP among active duty military personnel stationed in Hawaii.


2004 ◽  
Vol 85 (7) ◽  
pp. 2077-2085 ◽  
Author(s):  
P. Gilardi ◽  
I. García-Luque ◽  
M. T. Serra

In Capsicum, the resistance conferred by the L 2 gene is effective against all of the pepper-infecting tobamoviruses except Pepper mild mottle virus (PMMoV), whereas that conferred by the L 4 gene is effective against them all. These resistances are expressed by a hypersensitive response, manifested through the formation of necrotic local lesions (NLLs) at the primary site of infection. The Capsicum L 2 gene confers resistance to Paprika mild mottle virus (PaMMV), while the L 4 gene is effective against both PaMMV and PMMoV. The PaMMV and PMMoV coat proteins (CPs) were expressed in Capsicum frutescens (L 2 L 2) and Capsicum chacoense (L 4 L 4) plants using the heterologous Potato virus X (PVX)-based expression system. In C. frutescens (L 2 L 2) plants, the chimeric PVX virus containing the PaMMV CP was localized in the inoculated leaves and produced NLLs, whereas the chimeric PVX containing the PMMoV CP infected the plants systemically. Thus, the data indicated that the PaMMV CP is the only tobamovirus factor required for the induction of the host response mediated by the Capsicum L 2 resistance gene. In C. chacoense (L 4 L 4) plants, both chimeric viruses were localized to the inoculated leaves and produced NLLs, indicating that either PaMMV or PMMoV CPs are required to elicit the L 4 gene-mediated host response. In addition, transient expression of PaMMV CP into C. frutescens (L 2 L 2) leaves and PMMoV CP into C. chacoense (L 4 L 4) leaves by biolistic co-bombardment with a β-glucuronidase reporter gene led to the induction of cell death and the expression of host defence genes in both hosts. Thus, the tobamovirus CP is the elicitor of the Capsicum L 2 and L 4 gene-mediated hypersensitive response.


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