Search

Tomato mosaic disease caused by Tomato mosaic virus (ToMV) reduces tomato crop production globally. Biocontrol measures using various rhizobacteria and algae have been developed to reduce the adverse effects of plant diseases. To this end, two rhizobacteria (probiotic bacteria) including Pseudomonas fluorescens, Bacillus subtilis, and aqueous extract of brown alga (Sargassum angustifolium) were applied. A certain concentration of bacterial suspension was added to the tomato rhizosphere along with the aqueous extract of brown alga and the plants were subsequently inoculated with ToMV. Semi-quantitative indirect-ELISA was performed to estimate the virus titer within inoculated plants. Also, the disease severity index was determined by visual scoring of the plants at 14 and 28 days post-inoculation. Growth indices of plants were evaluated and the data were statistically analyzed. The results showed that multiple treatments of the rhizobacteria and the aqueous extract of brown alga reduced the disease severity to 27.46%, and inhibit the ToMV accumulation up to 86.48% in tomato plants. Moreover, the growth indices of tomato plants pre-treated with a combination of the rhizobacteria and brown alga extract were significantly improved. Taken together, the results suggest that these biocontrol agents have a synergistic effect and their simultaneous application can, therefore, reduce the crop loss caused by ToMV.

Earlier reports have indicated that begomoviruses encode four proteins (AC1/C1, AC2/C2, AC3/C3, and AC4/C4 proteins) using complementary‐sense DNA as the template. In recent years, several reports have shown that some begomoviruses also encode an AC5/C5 protein from the complementary DNA strand, and these AC5/C5 proteins play different roles in virus infections. Here, we provide evidence showing that Ageratum leaf curl Sichuan virus (ALCScV), a monopartite begomovirus, also encodes a C5 protein that is important for disease symptom formation and can affect viral replication. Infection of Nicotiana benthamiana plants with a potato virus X (PVX)‐based vector carrying the ALCScV C5 gene resulted in more severe disease symptoms and higher virus accumulation levels. ALCScV C5 protein can be found in the cytoplasm and the nucleus. Furthermore, this protein is also a suppressor of posttranscriptional gene silencing. Mutational analysis showed that knockout of C5 gene expression significantly reduced ALCScV‐induced disease symptoms and virus accumulation, while expression of the C5 gene using the PVX‐based vector enhanced ALCScV accumulation in coinfected N. benthamiana plants., ALCScV C5 protein is a virulence factor and enhances the pathogenicity of PVX in Nicotiana benthamiana. Mutational analyses showed that C5 is important for disease symptom formation and affects ALCScV replication in infected plants.

Cucurbit chlorotic yellows virus (CCYV) is a cucurbit-infecting crinivirus. RNA silencing can be initiated as a plant defense against viruses. Viruses encode various RNA silencing suppressors to counteract antiviral silencing. P22 protein encoded by RNA1 of CCYV is a silencing suppressor, but its mechanism of action remains unclear. In this study, the cucumber ribosomal-like protein CsRPS21 was found to interact with P22 protein in vitro and in vivo. A conserved CsRPS21 domain was indispensable for its nuclear localization and interaction with P22. Transient expression of CsRPS21 in Nicotiana benthamiana leaves interfered with P22 accumulation and inhibited P22 silencing suppressor activity. CsRPS21 expression in N. benthamiana protoplasts inhibited CCYV accumulation. Increasing numbers of ribosomal proteins are being found to be involved in viral infections of plants. We identified a P22-interacting ribosomal protein, CsRPS21, and uncovered its role in early viral replication and silencing suppressor activity. Our study increases knowledge of the function of ribosomal proteins during viral infection.

Plant viruses can be genetically modified to generate chimeric virus particles (CVPs) carrying heterologous peptides fused on the surface of coat protein (CP) subunits as vaccine candidates. However, some factors may be especially significant in determining the properties of chimeras. In this study, peptides from various sources and of various lengths were inserted into the Bamboo mosaic virus-based (BaMV) vector CP N-terminus to examine the chimeras infecting and accumulating in plants. Interestingly, it was found that the two different strains Foot-and-mouth disease virus (FMDV) VP1 antigens with flexible linker peptides (77 or 82 amino acids) were directly expressed on the BaMV CP, and the chimeric particles self-assembled and continued to express FMDV antigens. The chimeric CP, when directly fused with a large foreign protein (117 amino acids), can self-fold into incomplete virus particles or disks. The physicochemical properties of heterologus peptides N-terminus, complex strand structures of heterologus peptides C-terminus and different flexible linker peptides, can affect the chimera accumulation. Based on these findings, using plant virus-based chimeras to express foreign proteins can increase their length limitations, and engineered plant-made CVP-based vaccines have increasing potential for further development as novel vaccines.

Plant viruses can be genetically modified to generate chimeric virus particles (CVPs) carrying heterologous peptides fused on the surface of coat protein (CP) subunits as vaccine candidates. However, some factors may be especially significant in determining the properties of chimeras. In this study, peptides from various sources and of various lengths were inserted into the Bamboo mosaic virus-based (BaMV) vector CP N-terminus to examine the chimeras infecting and accumulating in plants. Interestingly, it was found that the two different strains Foot-and-mouth disease virus (FMDV) VP1 antigens with flexible linker peptides (77 or 82 amino acids) were directly expressed on the BaMV CP, and the chimeric particles self-assembled and continued to express FMDV antigens. The chimeric CP, when directly fused with a large foreign protein (117 amino acids), can self-fold into incomplete virus particles or disks. The physicochemical properties of heterologus peptides N-terminus, complex strand structures of heterologus peptides C-terminus and different flexible linker peptides, can affect the chimera accumulation. Based on these findings, using plant virus-based chimeras to express foreign proteins can increase their length limitations, and engineered plant-made CVP-based vaccines have increasing potential for further development as novel vaccines.

Rosellinia necatrix is responsible for the white rot root disease of avocado in Southern Spain. Entoleuca sp. is a fungus isolated from roots of these same trees, but it is not pathogenic in avocado. Here, we describe two new species of partitiviruses detected in isolates of the avocado sympatric fungi Entoleuca sp. and R. necatrix, termed Entoleuca partitivirus 1 (EnPV1), genus Alphapartitivirus, and Entoleuca partitivirus 2 (EnPV2), genus Betapartitivirus. For both R. necatrix and Entoleuca sp., the dsRNA of the RdRp genomic segment of EnPV1 accumulates at a higher rate than the CP dsRNA, except for a set of Entoleuca sp. isolates where titers of the CP dsRNA are 35–50 times higher than those of the RdRp dsRNA and between 250–380 times higher than the CP dsRNA titers found in the rest of Entoleuca sp. and R. necatrix isolates. For EnPV2, the accumulation rates of the RdRp dsRNA in Entoleuca sp., is in most of the cases, higher than the CP dsRNA. In contrast, in R. necatrix isolates, EnPV2 dsRNA2 generally accumulates at a higher rate. Genetic analysis of the partitiviruses revealed that there is no apparent variation in the nucleotide sequences among the strains. RNA silencing of the partitiviruses appears to be limited in Entoleuca sp., as shown by small RNA sequencing. Finally, the investigation of the presence of these partitiviruses in a fungal collection revealed that they have no role in the pathogenicity of R. necatrix in avocado or in the avirulence of Entoleuca sp. in this host., This work was supported by IFAPA project AVA201601.14, co-financed by FEDER, and by the Spanish Ministry of Science and Innovation project AGL2014-52518-C2-2-R.M.J. Alcalá technical assistance is acknowledged.

Thrips-transmitted tomato spotted wilt orthotospovirus (TSWV) causes spotted wilt disease in peanuts. A serological test (DAS-ELISA) is often used to detect TSWV in peanut leaf samples. However, in a few studies, DAS-ELISA detected more TSWV infection in root than leaf samples. It was not clear if the increased detection was due to increased TSWV accumulation in root tissue or merely an overestimation. Additionally, it was unclear if TSWV detection in asymptomatic plants would be affected by the detection technique. TSWV infection in leaf and root tissue from symptomatic and asymptomatic plants was compared via DAS-ELISA, RT-PCR, and RT-qPCR. TSWV incidence did not vary by DAS-ELISA, RT-PCR, and RT-qPCR in leaf and root samples of symptomatic plants or in leaf samples of asymptomatic plants. In contrast, significantly more TSWV infection and virus load were detected in root samples of asymptomatic plants via DAS-ELISA than other techniques suggesting that DAS-ELISA overestimated TSWV incidence and load. TSWV loads from symptomatic plants via RT-qPCR were higher in leaf than root samples, while TSWV loads in leaf and root samples from asymptomatic plants were not different but were lower than those in symptomatic plants. These findings suggested that peanut tissue type and detection technique could affect accurate TSWV detection and/or quantitation.

Tomato black ring virus (TBRV) is the only member of the Nepovirus genus that is known to form defective RNA particles (D RNAs) during replication. Here, de novo generation of D RNAs was observed during prolonged passages of TBRV isolates originated from Solanum lycopersicum and Lactuca sativa in Chenopodium quinoa plants. D RNAs of about 500 nt derived by a single deletion in the RNA1 molecule and contained a portion of the 5' untranslated region and viral replicase, and almost the entire 3' non-coding region. Short regions of sequence complementarity were found at the 5' and 3' junction borders, which can facilitate formation of the D RNAs. Moreover, in this study we analyzed the effects of D RNAs on TBRV replication and symptoms development of infected plants. C. quinoa, S. lycopersicum, Nicotiana tabacum, and L. sativa were infected with the original TBRV isolates (TBRV-D RNA) and those containing additional D RNA particles (TBRV + D RNA). The viral accumulation in particular hosts was measured up to 28 days post inoculation by RT-qPCR. Statistical analyses revealed that D RNAs interfere with TBRV replication and thus should be referred to as defective interfering particles. The magnitude of the interference effect depends on the interplay between TBRV isolate and host species., This work was supported by the National Science Centre, Poland (grant number 2017/25/B/NZ9/01715); Ministry of Science and Higher Education, Poland (grant number IP2014 014973) (to B.H.-J.) and Spain Agencia Estatal de Investigación-FEDER (grant number BFU2015-65037P) (to S.F.E.).

Virus diseases greatly affect oilseed rape (Brassica napus) production. Investigating antiviral genes may lead to the development of disease-resistant varieties of oilseed rape. In this study, we examined the effects of the suppressor of gene silencing 3 in Brassica napus (BnSGS3, a putative antiviral gene) with different genus viruses by constructing BnSGS3-overexpressing (BnSGS3-Ov) and BnSGS3-silenced (BnSGS3-Si) oilseed rape (cv. Zhongshuang No. 6) plants. These three viruses are Oilseed rape mosaic virus (ORMV), Turnip mosaic virus (TuMV) and Cucumber mosaic virus (CMV). The native BnSGS3 expressed in all examined tissues with the highest expression in siliques. All three viruses induced BnSGS3 expression, but ORMV induced a dramatic increase in the BnSGS3-Ov plants, followed by TuMV and CMV. Upon inoculation with three different viruses, transcript abundance of BnSGS3 gene follows: BnSGS3-Ov &gt, non-transgenic plants &gt, BnSGS3-Si. The accumulation quantities of ORMV and TuMV exhibited a similar trend. However, CMV accumulation showed an opposite trend where virus accumulations were negatively correlated with BnSGS3 expression. The results suggest that BnSGS3 selectively inhibits CMV accumulation but promotes ORMV and TuMV accumulation. BnSGS3 should be used in different ways (up- and down-regulation) for breeding virus-resistant oilseed rape varieties.

cet article est intégré dans un numéro spécial : Van de Waal, D. B., Elser, J. J., Martiny, A. C., Sterner, R. W., Cotner, J. B., eds (2018). Progress in Ecological Stoichiometry. Lausanne: Frontiers Media. doi: 10.3389/978-2-88945-621-5; International audience; Ecological stoichiometry and resource competition theory both predict that nutrient rates and ratios can alter infectious disease dynamics. Pathogens such as viruses hijack nutrient rich host metabolites to complete multiple steps of their epidemiological cycle. As the synthesis of these molecules requires nitrogen (N) and phosphorus (P), environmental supply rates, and ratios of N and P to hosts can directly limit disease dynamics. Environmental nutrient supplies also may alter virus epidemiology indirectly by changing host phenotype or the dynamics of coinfecting pathogens. We tested whether host nutrient supplies and coinfection control pathogen growth within hosts and transmission to new hosts, either directly or through modifications of plant tissue chemistry (i.e., content and stoichiometric ratios of nutrients), host phenotypic traits, or among-pathogen interactions. We examined two widespread plant viruses (BYDV-PAV and CYDV-RPV) in cultivated oats (Avena sativa) grown along a range of N and of P supply rates. N and P supply rates altered plant tissue chemistry and phenotypic traits; however, environmental nutrient supplies and plant tissue content and ratios of nutrients did not directly alter virus titer. Infection with CYDV-RPV altered plant traits and resulted in thicker plant leaves (i.e., higher leaf mass per area) and there was a positive correlation between CYDV-RPV titer and leaf mass per area. CYDV-RPV titer was reduced by the presence of a competitor, BYDV-PAV, and higher CYDV-RPV titer led to more severe chlorotic symptoms. In our experimental conditions, virus transmission was unaffected by nutrient supply rates, co-infection, plant stoichiometry, or plant traits, although nutrient supply rates have been shown to increase infection and coinfection rates. This work provides a robust test of the role of plant nutrient content and ratios in the dynamics of globally important pathogens and reveals a more complex relationship between within-host virus growth and alterations of plant traits. A deeper understanding of the differential effects of environmental nutrient supplies on virus epidemiology and ecology is particularly relevant given the rapid increase of nutrients flowing into Earth's ecosystems as a result of human activities.

La enfermedad de la hoja amarilla en caña de azúcar (SCYLV), es causada por el Sugarcane Yellow Leaf Virus (SCYLV) el cual pertenece al género Polerovirus de la familia Luteoviridae y es una enfermedad limitante en países productores de este cultivo debido a la disminución de parámetros de desarrollo y por ende en la productividad (biomasa y sacarosa). La detección de esta enfermedad a edad temprana en semilleros, es necesaria para la toma decisiones al momento del establecimiento de los cultivos comerciales; sin embargo, al contar con variedades asintomáticas al virus, se necesita el uso de herramientas de diagnóstico rápidas, confiables, de alta sensibilidad y cuantitativas. Este trabajo, a través de la técnica molecular RT PCR en tiempo real vía cuantificación absoluta, desarrolló un modelo para la cuantificación del número de copias de la carga viral a partir de un fragmento del virus. Con este modelo, se logró establecer las curvas de progreso de la enfermedad a través del tiempo de cultivo en cuatro variedades comerciales de alta importancia en Colombia: CC 01-1940, CC 84-75, CC 85- 92 y V 71-51 y se determinó su relación con variables de desarrollo y productividad. Adicional a esto, se compararon las metodologías actualmente empleadas en el país para el diagnóstico del SCYLV, demostrando el gran potencial de tiempo real. Finalmente, utilizando metodologías de secuenciación masiva, se determinaron los genomas del SCYLV presentes en estas variedades y se realizó el ensamblaje y comparación filogenética con la diversidad existente en otras regiones del mundo. //Abstract: Sugarcane Yellow Leaf Disease is caused by a virus with the same name (Sugarcane Yellow Leaf Virus, SCYLV), which belongs to the Polerovirus genus and Luteoviridae family. It is considered a limiting disease in many cane producing countries due to its negative impacts on crop development and diminishing yield and productivity (biomass and sugars). Detection of this disease at early stages, especially at nurseries, it is critical for decisions making about commercial cane crops establishment. However, because common occurrence of asymptomatic virus-diseased varieties, faster, reliable, highly sensitive, and as much as possible quantitative diagnostic techniques are required. In this study, by using the molecular technique Reverse Transcription - quantitative PCR (RT-qPCR) or real time PCR for absolute quantitation, a mathematical model was developed to quantify the number of copies for virus titer or viral load from a virus genome fragment. With this model, disease progress curves were established along the crop’s time course and development in four commercial cane varieties with high past and current importance for this agroindustry in Colombia: CC 01-1940, CC 84-75, CC 85- 92 y V 71-51. In addition, the relationship of viral titer with plant development and productivity variables was also established. On the other hand, SCYLV diagnostic methods currently used in Colombia were compared, demonstrating the significant advantage, high sensitivity capability and potential for RT-qPCR usage. Finally, by using next generation sequencing technology, de novo genomes for SCYLV isolates obtained from these four varieties were obtained and assembled, and alignments and phylogenetic analysis indicated that they were different from SCYLV isolates from other countries. Doctorado

[Background]: Pepino mosaic virus (PepMV) is an emerging plant pathogen that infects tomatoes worldwide. Understanding the factors that influence its evolutionary success is essential for developing new control strategies that may be more robust against the evolution of new viral strains. One of these evolutionary factors is the distribution of mutational fitness effect (DMFE), that is, the fraction of mutations that are lethal, deleterious, neutral, and beneficial on a given viral strain and host species. The goal of this study was to characterize the DMFE of introduced nonsynonymous mutations on a mild isolate of PepMV from the Chilean 2 strain (PepMV-P22). Additionally, we also explored whether the fitness effect of a given mutation depends on the gene where it appears or on epistatic interactions with the genetic background. To address this latter possibility, a subset of mutations were also introduced in a mild isolate of the European strain (PepMV-P11) and the fitness of the resulting clones measured., [Results]: A collection of 25 PepMV clones each containing a single nucleotide nonsynonymous substitution was created by site-directed mutagenesis and the fitness of each mutant was determined. PepMV-P22 genome showed a high degree of robustness against point mutations, with 80% of mutations being either neutral or even beneficial and only 20% being deleterious or lethal. We found that the effect of mutations strongly depended on the gene in which they were introduced. Mutations with the largest average beneficial effects were those affecting the RdRp gene, in contrast to mutations affecting TGB1 and CP genes, for which the average effects were deleterious. Moreover, significant epistatic interactions were observed between nonsynonymous mutations and the genetic background, meaning that the effect of a given nucleotide substitution on a particular genomic context cannot be predicted by knowing its effect in a different one., [Conclusions]: Our results indicated that PepMV genome has a surprisingly high robustness against mutations. We also found that fitness consequences of a given mutation differ between the two strains analyzed. This discovery suggests that the strength of selection, and thus the rates of evolution, vary among PepMV strains., This study was financially supported by grant 2011/01/D/NZ9/00279, from the Poland National Science Center, to B.H.J and by grants BFU2015-65037-P, from Spain Ministry of Economy and Competitiveness-FEDER, and PROMETEOII/2014/021, from Generalitat Valenciana, to S.F.E.

Summary � The viral genome-linked protein, VPg, of potyviruses is involved in viral genome replication and translation. To determine host proteins that interact with Sugarcane mosaic virus (SCMV) VPg, a yeast two-hybrid screen was used and a maize (Zea mays) Elongin C (ZmElc) protein was identified. � ZmELC transcript was observed in all maize organs, but most highly in leaves and pistil extracts, and ZmElc was present in the cytoplasm and nucleus of maize cells in the presence or absence of SCMV. ZmELC expression was increased in maize tissue at 4 and 6 d post SCMV inoculation. When ZmELC was transiently overexpressed in maize protoplasts the accumulation of SCMV RNA was approximately doubled compared with the amount of virus in control protoplasts. � Silencing ZmELC expression using a Brome mosaic virus-based gene silencing vector (virusinduced gene silencing) did not influence maize plant growth and development, but did decrease RNA accumulation of two isolates of SCMV and host transcript encoding ZmeIF4E during SCMV infection. Interestingly, Maize chlorotic mottle virus, from outside the Potyviridae, was increased in accumulation after silencing ZmELC expression. � Our results describe both the location of ZmElc expression in maize and a new activity associated with an Elc: support of potyvirus accumulation.

Plant viruses use cellular factors and resources to replicate and move. Plants respond to viral infection by several mechanisms, including innate immunity, autophagy, and gene silencing, that viruses must evade or suppress. Thus, the establishment of infection is genetically determined by the availability of host factors necessary for virus replication and movement and by the balance between plant defense and viral suppression of defense responses. Host factors may have antiviral or proviral activities. Proviral factors condition susceptibility to viruses by participating in processes essential to the virus. Here, we review current advances in the identification and characterization of host factors that condition susceptibility to plant viruses. Host factors with proviral activity have been identified for all parts of the virus infection cycle: viral RNA translation, viral replication complex formation, accumulation or activity of virus replication proteins, virus movement, and virion assembly. These factors could be targets of gene editing to engineer resistance to plant viruses.

Background Production of cassava (Manihot esculenta Crantz), a food security crop in sub-Saharan Africa, is threatened by the spread of cassava brown streak disease (CBSD) which manifests in part as a corky necrosis in the storage root. It is caused by either of two virus species, Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV), resulting in up to 100% yield loss in susceptible varieties. Methods This study characterized the response of 11 cassava varieties according to CBSD symptom expression and relative CBSV and UCBSV load in a field trial in Uganda. Relative viral load was measured using quantitative RT-PCR using COX as an internal housekeeping gene. Results A complex situation was revealed with indications of different resistance mechanisms that restrict virus accumulation and symptom expression. Four response categories were defined. Symptom expression was not always positively correlated with virus load. Substantially different levels of the virus species were found in many genotypes suggesting either resistance to one virus species or the other, or some form of interaction, antagonism or competition between virus species. Conclusions A substantial amount of research still needs to be undertaken to fully understand the mechanism and genetic bases of resistance. This information will be useful in informing breeding strategies and restricting virus spread. Electronic supplementary material The online version of this article (doi:10.1186/s12985-014-0216-x) contains supplementary material, which is available to authorized users.

) appear to be highly virulent for chickens. The chance of AFV infection of chickens in case of intranasal challenge is 20 times as great as in the case of peroral one, and 300 times as great as in the case of intragastral one, which bears evidence to higher sensitivity to AFV of the tissues of avian respiratory organs, in comparison with the tissues of gastro-intestinal tract. Therewith, primary target organ for virus in intranasal infected birds is their respiratory channel (mucous membrane of the nasal cavity in particular). Registered is the possibility of existence of fecal-nasal AFV transfer mechanism in chickens.

Nicotiana benthamiana plants were engineered to express sequences of the helper component-proteinase (HC-Pro) of Cowpea aphid-borne mosaic potyvirus (CABMV). The sensitivity of the transgenic plants to infection with parental and heterologous viruses was studied. The lines expressing HC-Pro showed enhanced symptoms after infection with the parental CABMV isolate and also after infection with a heterologous potyvirus, Potato virus Y (PVY) and a comovirus, Cowpea mosaic virus (CPMV). On the other hand, transgenic lines expressing nontranslatable HC-Pro or translatable HC-Pro with a deletion of the central domain showed wild type symptoms after infection with the parental CABMV isolate and heterologous viruses. These results showed that CABMV HC-Pro is a pathogenicity determinant that conditions enhanced sensitivity to virus infection in plants, and that the central domain of the protein is essential for this. The severe symptoms in CABMV-infected HC-Pro expressing lines were remarkably followed by brief recovery and subsequent re-establishment of infection, possibly indicating counteracting effects of HC-Pro expression and a host defense response. One of the HC-Pro expressing lines (h48) was found to contain low levels of transgenic HC-Pro RNA and to be resistant to CABMV and to recombinant CPMV expressing HC-Pro. This indicated that h48 was (partially) posttranscriptionally silenced for the HC-Pro transgene inspite of the established role of HC-Pro as a suppressor of posttranscriptional gene silencing. Line h48 was not resistant to PVY, but instead showed enhanced symptoms compared to nontransgenic plants. This may be due to relief of silencing of the HC-Pro transgene by HC-Pro expressed by PVY.