Preliminary Study of Viruses Infecting Strawberry (Fragaria spp.) in the Mid-Southern United States

Strawberry is an important economic crop in the mid-southern United States, however, limited information is known about the viruses present in strawberry fields in Arkansas, Missouri, Oklahoma, and Texas. The purpose of this study was to determine the presence of particular viruses infecting strawberry crops in the above four states. A total of 816 strawberry leaf samples were randomly collected from grower’s fields during the 2014 and 2016 growing seasons and tested by dot-immunobinding assay (DIBA) against antisera of eight available viruses: apple mosaic virus (ApMV), arabis mosaic virus (ArMV), raspberry bushy dwarf virus (RBDV), raspberry ringspot virus (RpRSV), strawberry mild yellow edge virus (SMYEV), tobacco necrosis virus (TNV), tomato black ring virus (TBRV), and tomato ringspot virus (ToRSV). Of the eight viruses, ApMV, RBDV, and SMYEV had the highest infection rates in 2014, while ApMV, ArMV, ToRSV and SMYEV were the most prevalent viruses in 2016. Mixed infection of more than one virus was very common in samples collected from all four states. Our preliminary results showed that the distribution of virus relative frequencies differed significantly among seven sample locations across the states within a sample year, but we could not show statistical differences among the years based on our sample sizes. Mixed infection in strawberries were higher than expected in 2014 and lower than expected in 2016 based on viruses being independent infections. Overall, the occurrence of the viruses in strawberry crops could have important potential impacts for strawberry production in the mid-southern states.

The Structural Characterization and Antipathogenic Activities of Quinoin, a Type 1 Ribosome-Inactivating Protein from Quinoa Seeds

Quinoin is a type 1 ribosome-inactivating protein (RIP) we previously isolated from the seeds of pseudocereal quinoa (Chenopodium quinoa) and is known as a functional food for its beneficial effects on human health. As the presence of RIPs in edible plants could be potentially risky, here we further characterised biochemically the protein (complete amino acid sequence, homologies/differences with other RIPs and three-dimensional homology modeling) and explored its possible defensive role against pathogens. Quinoin consists of 254 amino acid residues, without cysteinyl residues. As demonstrated by similarities and homology modeling, quinoin preserves the amino acid residues of the active site (Tyr75, Tyr122, Glu177, Arg180, Phe181 and Trp206; quinoin numbering) and the RIP-fold characteristic of RIPs. The polypeptide chain of quinoin contains two N-glycosylation sites at Asn115 and Asp231, the second of which appears to be linked to sugars. Moreover, by comparative MALDI-TOF tryptic peptide mapping, two differently glycosylated forms of quinoin, named pre-quinoin-1 and pre-quinoin-2 (~0.11 mg/100 g and ~0.85 mg/100 g of seeds, respectively) were characterised. Finally, quinoin possesses: (i) strong antiviral activity, both in vitro and in vivo towards Tobacco Necrosis Virus (TNV); (ii) a growth inhibition effect on the bacterial pathogens of plants; and (iii) a slight antifungal effect against two Cryphonectria parasitica strains.

Frequencies 171.160 Hz, 174.989 Hz, and 1185.131 Hz useful in combating COVID-19 (Novel Coronavirus)

The paper presents the works carried out by different researchers for identifying the SARS-CoV-2 virus structure, and its background with the present medication being addressed to the COVID-19 patients. The paper discusses three candidates for possible defense and cure of humans against the virus along with the regular medication. The first is the resonant frequency therapy, which has proven its efficacy for treating different types of viral infection and cancer cells as the capsid of Rubella measles virus, Satellite Tobacco necrosis Virus, Cancer cells and many more are neutralized on the application of frequency equal to the resonance frequency of the protein shell of the virus. It is expected that there must be a resonance frequency corresponding to the capsid of the SARS-CoV-2, on the application of which might result in the disintegration of the protein shell. As treating a huge number of people in hospitals is becoming increasingly difficult and large number of population is under quarantine, a resonance audio frequency of this sort embedded in a soothing music can be melodius, and the same if played in public spaces and homes might help a great deal. The frequencies (approximate values) are 171.160 Hz, 174.989 Hz, 1185.131 Hz the complete details regarding administration and calculation of the frequencies are discussed in this manuscript. This treatment along with the presently administered treatment to COVID-19 patients might aid in their quick recovery from illness. Further the audio frequency/ resonance frequency therapy, and the strategy discussed in this manuscript might be useful in future from any kind of Virus attacks/ Biological warfares either intentionally or naturally. Also, the medicinal properties of ethnomedical plants such as Piper nigrum, and Ocimum tenuiflorum L and essential oils such as Eucalyptus oil are discussed. The last one being the antiviral activities of copper in the form of copper oxide nanoparticles in medicine which has proven its efficacy for viral prevention, replication, and RNA degradation are also discussed.

Emerging novel Coronavirus: a review of three potential candidates that probably debilitate the proliferation of the virus

The paper presents the works carried out by different researchers for identifying the SARS-CoV-2 virus structure, and its background with the present medication being addressed to the COVID-19 patients. The paper discusses three candidates for possible defense and cure of humans against the virus along with the regular medication. The first is the resonant frequency therapy, which has proven its efficacy for treating different types of viral infection and cancer cells as the capsid of Rubella measles virus, Satellite Tobacco necrosis Virus, Cancer cells and many more are neutralized on the application of frequency equal to the resonance frequency of the protein shell of the virus. It is expected that there must be a resonance frequency corresponding to the capsid of the SARS-CoV-2, on the application of which might result in the disintegration of the protein shell. As treating a huge number of people in hospitals is becoming increasingly difficult and large number of population is under quarantine, a resonance audio frequency of this sort embedded in a soothing music can be melodius, and the same if played in public spaces and homes might help a great deal. The frequencies (approximate values) are 171.160 Hz, 174.989 Hz, 1185.131 Hz the complete details regarding administration and calculation of the frequencies are discussed in this manuscript. This treatment along with the presently administered treatment to COVID-19 patients might aid in their quick recovery from illness. Further the audio frequency/ resonance frequency therapy, and the strategy discussed in this manuscript might be useful in future from any kind of Virus attacks/ Biological warfares either intentionally or naturally. Also, the medicinal properties of ethnomedical plants such as Piper nigrum, and Ocimum tenuiflorum L and essential oils such as Eucalyptus oil are discussed. The last one being the antiviral activities of copper in the form of copper oxide nanoparticles in medicine which has proven its efficacy for viral prevention, replication, and RNA degradation are also discussed.

Tobacco Necrosis Virus-AC Single Coat Protein Amino Acid Substitutions Determine Host-Specific Systemic Infections of Nicotiana benthamiana and Soybean

Plant viruses often infect several distinct host species. Sometimes, viruses can systemically infect a specific host whereas, in other cases, only local infections occur in other species. How viral and host factors interact to determine systemic infections among different hosts is largely unknown, particularly for icosahedral positive-stranded RNA viruses. The Tobacco necrosis virus-A Chinese isolate belongs to the genus Alphanecrovirus in the family Tombusviridae. In this study, we investigated variations in systemic infections of tobacco necrosis virus-AC (TNV-AC) in Nicotiana benthamiana and Glycine max (soybean) by alanine-scanning mutagenesis of the viral coat protein (CP), which is essential for systemic movement of TNV-AC. We found that three amino acids, R169, K177, and Q233, are key residues that mediate varying degrees of systemic infections of N. benthamiana and soybean. Further analysis revealed that variations in systemic trafficking of TNV-AC CP mutants in N. benthamiana and soybean are associated with virion assembly and stability. The CP amino acids K177 and Q233 are highly conserved among all TNV-A isolates and are replaced by Q and K in the TNV-D isolates. We demonstrated that systemic infectivity of either TNV-AC K177A and Q233A or K177Q and Q233K mutants are correlated with the binding affinity of the mutated CPs to the host-specific Hsc70-2 protein. These results expand our understanding of host-dependent long-distance movement of icosahedral viruses in plants. [Formula: see text] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Antiviral Activity of PD-L1 and PD-L4, Type 1 Ribosome Inactivating Proteins from Leaves of Phytolacca dioica L. in the Pathosystem Phaseolus vulgaris–Tobacco Necrosis Virus (TNV)

Using the pathosystem Phaseolus vulgaris–tobacco necrosis virus (TNV), we demonstrated that PD-L1 and PD-L4, type-1 ribosome inactivating proteins (RIPs) from leaves of Phytolacca dioica L., possess a strong antiviral activity. This activity was exerted both when the RIPs and the virus were inoculated together in the same leaf and when they were inoculated or applied separately in the adaxial and abaxial leaf surfaces. This suggests that virus inhibition would mainly occur inside plant cells at the onset of infection. Histochemical studies showed that both PD-L1 and PD-L4 were not able to induce oxidative burst and cell death in treated leaves, which were instead elicited by inoculation of the virus alone. Furthermore, when RIPs and TNV were inoculated together, no sign of H2O2 deposits and cell death were detectable, indicating that the virus could have been inactivated in a very early stage of infection, before the elicitation of a hypersensitivity reaction. In conclusion, the strong antiviral activity is likely exerted inside host cells as soon the virus disassembles to start translation of the viral genome. This activity is likely directed towards both viral and ribosomal RNA, explaining the almost complete abolition of infection when virus and RIP enter together into the cells.

Investigation of Novel RNA Elements in the 3′UTR of Tobacco Necrosis Virus-D

RNA elements in the untranslated regions of plus-strand RNA viruses can control a variety of viral processes including translation, replication, packaging, and subgenomic mRNA production. The 3′ untranslated region (3′UTR) of Tobacco necrosis virus strain D (TNV-D; genus Betanecrovirus, family Tombusviridae) contains several well studied regulatory RNA elements. Here, we explore a previously unexamined region of the viral 3′UTR, the sequence located upstream of the 3′-cap independent translation enhancer (3′CITE). Our results indicate that (i) a long-range RNA–RNA interaction between an internal RNA element and the 3′UTR facilitates translational readthrough, and may also promote viral RNA synthesis; (ii) a conserved RNA hairpin, SLX, is required for efficient genome accumulation; and (iii) an adenine-rich region upstream of the 3′CITE is dispensable, but can modulate genome accumulation. These findings identified novel regulatory RNA elements in the 3′UTR of the TNV-D genome that are important for virus survival.

RNAseq Analysis of Rhizomania-Infected Sugar Beet Provides the First Genome Sequence of Beet Necrotic Yellow Vein Virus from the USA and Identifies a Novel Alphanecrovirus and Putative Satellite Viruses

“Rhizomania” of sugar beet is a soilborne disease complex comprised of beet necrotic yellow vein virus (BNYVV) and its plasmodiophorid vector, Polymyxa betae. Although BNYVV is considered the causal agent of rhizomania, additional viruses frequently accompany BNYVV in diseased roots. In an effort to better understand the virus cohort present in sugar beet roots exhibiting rhizomania disease symptoms, five independent RNA samples prepared from diseased beet seedlings reared in a greenhouse or from field-grown adult sugar beet plants and enriched for virus particles were subjected to RNAseq. In all but a healthy control sample, the technique was successful at identifying BNYVV and provided sequence reads of sufficient quantity and overlap to assemble > 98% of the published genome of the virus. Utilizing the derived consensus sequence of BNYVV, infectious RNA was produced from cDNA clones of RNAs 1 and 2. The approach also enabled the detection of beet soilborne mosaic virus (BSBMV), beet soilborne virus (BSBV), beet black scorch virus (BBSV), and beet virus Q (BVQ), with near-complete genome assembly afforded to BSBMV and BBSV. In one field sample, a novel virus sequence of 3682 nt was assembled with significant sequence similarity and open reading frame (ORF) organization to members within the subgenus Alphanecrovirus (genus Necrovirus; family Tombusviridae). Construction of a DNA clone based on this sequence led to the production of the novel RNA genome in vitro that was capable of inducing local lesion formation on leaves of Chenopodium quinoa. Additionally, two previously unreported satellite viruses were revealed in the study; one possessing weak similarity to satellite maize white line mosaic virus and a second possessing moderate similarity to satellite tobacco necrosis virus C. Taken together, the approach provides an efficient pipeline to characterize variation in the BNYVV genome and to document the presence of other viruses potentially associated with disease severity or the ability to overcome resistance genes used for sugar beet rhizomania disease management.

Emerging novel Coronavirus: a review of three potential candidates that probably debilitate the proliferation of the virus

The review paper presents the works carried out by different researchers for identifying the SARS-CoV-2 virus structure, and its background with the present medication being addressed to the COVID-19 patients. The paper discusses three candidates for possible defense and cure of humans against the virus along with the regular medication. The first is the resonant frequency therapy, which has proven its efficacy for treating different types of viral infection and cancer cells as the capsid of Rubella measles virus, Satellite Tobacco necrosis Virus, Cancer cells and many more are neutralized on the application of frequency equal to the resonance frequency of the protein shell of the virus. It is expected that there must be a resonance frequency corresponding to the capsid of the SARS-CoV-2, on the application of which might result in the disintegration of the protein shell. As treating a huge number of people in hospitals is becoming increasingly difficult and large number of population is under quarantine, a resonance audio frequency of this sort embedded in a soothing music can be melodius, and the same if played in public spaces and homes might help a great deal. The frequencies (approximate values) are 171.160 Hz, 174.989 Hz, 1185.131 Hz the complete details regarding administration and calculation of the frequencies are discussed in this manuscript. This treatment along with the presently administered treatment to COVID-19 patients might aid in their quick recovery from illness. Further the audio frequency/ resonance frequency therapy, and the strategy discussed in this manuscript might be useful in future from any kind of Virus attacks/ Biological warfares either intentionally or naturally. Also, the medicinal properties of ethnomedical plants such as Piper nigrum, and Ocimum tenuiflorum L and essential oils such as Eucalyptus oil are discussed. The last one being the antiviral activities of copper in the form of copper oxide nanoparticles in medicine which has proven its efficacy for viral prevention, replication, and RNA degradation are also discussed.

Emerging novel Coronavirus: a review of three potential candidates that probably debilitate the proliferation of the virus

The review paper presents the works carried out by different researchers for identifying the SARS-CoV-2 virus structure, and its background with the present medication being addressed to the COVID-19 patients. The paper discusses three candidates for possible defense and cure of humans against the virus along with the regular medication. The first is the resonant frequency therapy, which has proven its efficacy for treating different types of viral infection and cancer cells as the capsid of Rubella measles virus, Satellite Tobacco necrosis Virus, Cancer cells and many more are neutralized on the application of frequency equal to the resonance frequency of the protein shell of the virus. It is expected that there must be a resonance frequency corresponding to the capsid of the SARS-CoV-2, on the application of which might result in the disintegration of the protein shell. As treating a huge number of people in hospitals is becoming increasingly difficult and large number of population is under quarantine, a resonance audio frequency of this sort embedded in a soothing music can be melodius, and the same if played in public spaces and homes might help a great deal. The frequencies (approximate values) are 171.160 Hz, 174.989 Hz, 1185.131 Hz the complete details regarding administration and calculation of the frequencies are discussed in this manuscript. This treatment along with the presently administered treatment to COVID-19 patients might aid in their quick recovery from illness. Further the audio frequency/ resonance frequency therapy, and the strategy discussed in this manuscript might be useful in future from any kind of Virus attacks/ Biological warfares either intentionally or naturally. Also, the medicinal properties of ethnomedical plants such as Piper nigrum, and Ocimum tenuiflorum L and essential oils such as Eucalyptus oil are discussed. The last one being the antiviral activities of copper in the form of copper oxide nanoparticles in medicine which has proven its efficacy for viral prevention, replication, and RNA degradation are also discussed.