Correction: Estimation of the dispersal distances of an aphid-borne virus in a patchy landscape.

[This corrects the article DOI: 10.1371/journal.pcbi.1006085.].

Estimation of the dispersal distances of an aphid-borne virus in a patchy landscape.

Characterising the spatio-temporal dynamics of pathogens in natura is key to ensuring their efficient prevention and control. However, it is notoriously difficult to estimate dispersal parameters at scales that are relevant to real epidemics. Epidemiological surveys can provide informative data, but parameter estimation can be hampered when the timing of the epidemiological events is uncertain, and in the presence of interactions between disease spread, surveillance, and control. Further complications arise from imperfect detection of disease and from the huge number of data on individual hosts arising from landscape-level surveys. Here, we present a Bayesian framework that overcomes these barriers by integrating over associated uncertainties in a model explicitly combining the processes of disease dispersal, surveillance and control. Using a novel computationally efficient approach to account for patch geometry, we demonstrate that disease dispersal distances can be estimated accurately in a patchy (i.e. fragmented) landscape when disease control is ongoing. Applying this model to data for an aphid-borne virus (Plum pox virus) surveyed for 15 years in 605 orchards, we obtain the first estimate of the distribution of flight distances of infectious aphids at the landscape scale. About 50% of aphid flights terminate beyond 90 m, which implies that most infectious aphids leaving a tree land outside the bounds of a 1-ha orchard. Moreover, long-distance flights are not rare-10% of flights exceed 1 km. By their impact on our quantitative understanding of winged aphid dispersal, these results can inform the design of management strategies for plant viruses, which are mainly aphid-borne.

Analyzing the Influence of Landscape Aggregation on Disease Spread to Improve Management Strategies.

Epidemiological models are increasingly used to predict epidemics and improve management strategies. However, they rarely consider landscape characteristics although such characteristics can influence the epidemic dynamics and, thus, the effectiveness of disease management strategies. Here, we present a generic in silico approach which assesses the influence of landscape aggregation on the costs associated with an epidemic and on improved management strategies. We apply this approach to sharka, one of the most damaging diseases of Prunus trees, for which a management strategy is already applied in France. Epidemic simulations were carried out with a spatiotemporal stochastic model under various management strategies in landscapes differing in patch aggregation. Using sensitivity analyses, we highlight the impact of management parameters on the economic output of the model. We also show that the sensitivity analysis can be exploited to identify several strategies that are, according to the model, more profitable than the current French strategy. Some of these strategies are specific to a given aggregation level, which shows that management strategies should generally be tailored to each specific landscape. However, we also identified a strategy that is efficient for all levels of landscape aggregation. This one-size-fits-all strategy has important practical implications because of its simple applicability at a large scale.

Improving Management Strategies of Plant Diseases Using Sequential Sensitivity Analyses.

Improvement of management strategies of epidemics is often hampered by constraints on experiments at large spatiotemporal scales. A promising approach consists of modeling the biological epidemic process and human interventions, which both impact disease spread. However, few methods enable the simultaneous optimization of the numerous parameters of sophisticated control strategies. To do so, we propose a heuristic approach (i.e., a practical improvement method approximating an optimal solution) based on sequential sensitivity analyses. In addition, we use an economic improvement criterion based on the net present value, accounting for both the cost of the different control measures and the benefit generated by disease suppression. This work is motivated by sharka (caused by Plum pox virus), a vector-borne disease of prunus trees (especially apricot, peach, and plum), the management of which in orchards is mainly based on surveillance and tree removal. We identified the key parameters of a spatiotemporal model simulating sharka spread and control and approximated optimal values for these parameters. The results indicate that the current French management of sharka efficiently controls the disease, but it can be economically improved using alternative strategies that are identified and discussed. The general approach should help policy makers to design sustainable and cost-effective strategies for disease management.

Assessing the Mismatch Between Incubation and Latent Periods for Vector-Borne Diseases: The Case of Sharka.

The relative durations of the incubation period (the time between inoculation and symptom expression) and of the latent period (the time between inoculation and infectiousness of the host) are poorly documented for plant diseases. However, the extent of asynchrony between the ends of these two periods (i.e., their mismatch) can be a key determinant of the epidemic dynamics for many diseases and consequently it is of primary interest in the design of disease management strategies. In order to assess this mismatch, an experimental approach was developed and applied using sharka, a severe disease caused by Plum pox virus (PPV, genus Potyvirus, family Potyviridae) affecting trees belonging to the genus Prunus. Leaves of infected young peach trees were used individually as viral sources in aphid-mediated transmission tests carried out at different time points postinoculation in order to bracket symptom onset. By fitting a nonlinear logistic model to the obtained transmission rates, we demonstrated that the first symptoms appear on leaves 1 day before they rapidly become infectious. In addition, among symptomatic leaves, symptom intensity and transmission rate are positively correlated. These results strengthen the conclusion that, under our experimental conditions, incubation and latent periods of PPV infection are almost synchronous.

Surface plasmon resonance for monitoring the interaction of Potato virus Y with monoclonal antibodies.

Surface plasmon resonance (SPR)-based biosensors have been widely utilized for measuring interactions of a variety of molecules. Fewer examples include higher biological entities such as bacteria and viruses, and even fewer deal with plant viruses. Here, we describe the optimization of an SPR sensor chip for evaluation of the interaction of the economically relevant filamentous Potato virus Y (PVY) with monoclonal antibodies. Different virus isolates were efficiently and stably bound to a previously immobilized polyclonal antibody surface, which remained stable over subsequent injection regeneration steps. The ability of the biosensor to detect and quantify PVY particles was compared with ELISA and RT-qPCR. Stably captured virus surfaces were successfully used to explore kinetic parameters of the interaction of a panel of monoclonal antibodies with two PVY isolates representing the main viral serotypes N and O. In addition, the optimized biosensor proved to be suitable for evaluating whether two given monoclonal antibodies compete for the same epitope within the viral particle surface. The strategy proposed in this work can help to improve existing serologic diagnostic tools that target PVY and will allow investigation of the inherent serological variability of the virus and exploration for new interactions of PVY particles with other proteins.

Asymmetric interactions between barley yellow dwarf virus -PAV and wheat dwarf virus in wheat.

The deciphering of the epidemiology of a plant virus has long been focused on the study of interactions between partners of one pathosystem. However, plants are exposed to numerous viruses which lead to frequent co-infection scenarios. This can change characteristics of virus-vector-host interactions and could impact the epidemiology of viral diseases. Barley yellow dwarf virus-PAV (BYDV-PAV; species: Luteovirus pavhordei; genus Luteovirus), wheat dwarf virus (WDV; genus Mastrevirus) and their respective vectors (BYDV-PAV: e.g. Rhopalosiphum padi and WDV: Psammotettix alienus) are commonly found in cereal fields. Wheat plants co-infected with BYDV-PAV and WDV have been reported from field surveys, although epidemiological outcomes of BYDV-PAV - WDV interactions in planta have not yet been studied. Experiments were carried out to evaluate and compare, through different competition scenarios (i.e. single- and co- (simultaneous and sequential) inoculations), the efficiency of BYDV-PAV and WDV to infect, to accumulate in and to be spread between wheat plants. Moreover, the impact of competition scenarios on the biological parameters of these two viruses was evaluated at different stages of the infection and with plants at different ages at inoculation. Results showed i) that these viruses achieve their infection cycle and their plant-to-plant transmission with different efficiencies and ii) BYDV-PAV - WDV interactions lead to different phenotypes ranging from antagonism to synergism. Finally, when these two viruses share a host, the nature and strength of virus-virus interactions varied depending on the order of virus arrival, stages of the infection cycle and plant age at inoculation. Precisely, the introduction (i.e. co- and sequential inoculation) and infection process (i.e. virus accumulation) of BYDV-PAV in a wheat benefit from the presence of WDV. For the latter, the sympatry with BYDV-PAV exerts opposite pressure on parameters involved in virus introduction (i.e. benefit during sequential inoculation) and spread (i.e. lower transmission efficiency and virus accumulation in co-infected plants). In the context of increased potential exposure of crops to insect vectors, this study participates in a better understanding of the impact of BYDV-PAV and WDV co-infections on biological and ecological parameters of the diseases induced by these viruses.

Molecular evolution and phylogeography of potato virus Y based on the CP gene.

Potato virus Y (PVY) is an important plant pathogen with a wide host range that includes, among others, potato, tobacco, tomato and pepper. The coat protein (CP) of PVY has been commonly used in phylogenetic studies for strain classification. In this study, we used a pool of 292 CP sequences from isolates collected worldwide. After detecting and removing recombinant sequences, we applied Bayesian techniques to study the influence of geography and host species in CP population structure and dynamics. Finally, we performed selection and covariation analyses to identify specific amino acids involved in adaptation. Our results show that PVY CP diversification is significantly accounted for by both geographical and host-driven adaptations. Amino acid positions detected as positively selected concentrate in the N-terminal region of the protein. Some of these selected positions may discriminate among strains, and to a much lesser extent, between potato and non-potato isolates.

Can Mixed Intercropping Protect Cereals from Aphid-Borne Viruses? An Experimental Approach.

Intercropping, i.e., association of two or more species, is promising to reduce insect populations in fields. The cereal aphid Rhopalosiphum padi, a vector of the Barley yellow dwarf virus PAV (BYDV-PAV), represents a major threat for cereal grain production. In this study, we tested the potential of winter barley intercropped with clover to reduce the size of R. padi populations and to lower the BYDV-PAV incidence in fields. We used arenas (i.e., sets of 36 barley plants) intercropped with or without clover plants (at different sown densities). In each arena, a single viruliferous founder, R. padi, (with an alate or a wingless morph) was deposited to introduce aphids and viruses in the experiment. Thirteen days later, the number of aphids in the arena, the percentage of plants hosting aphids and the infection rates were monitored. Data produced through this experimental design showed that clover alters the distribution of the aphid progeny (lower aphid spread) produced by an alate founder morph. Moreover, clover reduces the size of aphid populations produced by a wingless founder morph. However, despite the effects of clover on biological parameters of R. padi, the presence of clover in barley arena did not modify BYDV infections, suggesting complex mechanisms between partners of the BYDV pathosystem for plant-to-plant virus spread.

The helper component proteinase cistron of Potato virus Y induces hypersensitivity and resistance in Potato genotypes carrying dominant resistance genes on chromosome IV.

The Nc(tbr) and Ny(tbr) genes in Solanum tuberosum determine hypersensitive reactions, characterized by necrotic reactions and restriction of the virus systemic movement, toward isolates belonging to clade C and clade O of Potato virus Y (PVY), respectively. We describe a new resistance from S. sparsipilum which possesses the same phenotype and specificity as Nc(tbr) and is controlled by a dominant gene designated Nc(spl). Nc(spl) maps on potato chromosome IV close or allelic to Ny(tbr). The helper component proteinase (HC-Pro) cistron of PVY was shown to control necrotic reactions and resistance elicitation in plants carrying Nc(spl), Nc(tbr), and Ny(tbr). However, inductions of necrosis and of resistance to the systemic virus movement in plants carrying Nc(spl) reside in different regions of the HC-Pro cistron. Also, genomic determinants outside the HC-Pro cistron are involved in the systemic movement of PVY after induction of necroses on inoculated leaves of plants carrying Ny(tbr). These results suggest that the Ny(tbr) resistance may have been involved in the recent emergence of PVY isolates with a recombination breakpoint near the junction of HC-Pro and P3 cistrons in potato crops. Therefore, this emergence could constitute one of the rare examples of resistance breakdown by a virus which was caused by recombination instead of by successive accumulation of nucleotide substitutions.

Fine Characterization of a Resistance Phenotype by Analyzing TuYV-Myzus persicae-Rapeseed Interactions.

Turnip yellows virus (TuYV), transmitted by Myzus persicae, can be controlled in rapeseed fields by insecticide treatments. However, the recent ban of the neonicotinoids together with the description of pyrethrinoid-resistant aphids has weakened insecticide-based control methods available to farmers. Since the deployment of insecticides in the 1980s, few research efforts were made to breed for rapeseed cultivars resistant to aphid-borne viral diseases. Thus, only few rapeseed cultivars released in Europe were reported to be TuYV-resistant, and the resistance phenotype of these cultivars was poorly characterized. In this study, several epidemiological parameters (infection rate, latency period, etc.) associated to the TuYV-resistance of the cv. Architect were estimated. Results showed a partial resistance phenotype for plants inoculated at the 2-/4-leaves stages and a resistance phenotype for plants inoculated at a more advanced growing stage. Moreover, analysis of infected plants highlighted (i) a poor quality of infected cv. Architect as a source of virus for transmission and (ii) an extended latency period for infected plants. Thus, dynamics of virus spread in the field should to be slower for Architect compared to susceptible rapeseed cultivars, which should lead to the maintenance of a higher proportion of healthy plants in the field.

Efficiency and Persistence of Movento® Treatment against Myzus persicae and the Transmission of Aphid-Borne Viruses.

Neonicotinoids are widely used to protect fields against aphid-borne viral diseases. The recent ban of these chemical compounds in the European Union has strongly impacted rapeseed and sugar beet growing practices. The poor sustainability of other insecticide families and the low efficiency of prophylactic methods to control aphid populations and pathogen introduction strengthen the need to characterize the efficiency of new plant protection products targeting aphids. In this study, the impact of Movento® (Bayer S.A.S., Leverkusen, Germany), a tetrameric acid derivative of spirotetramat, on Myzus persicae and on viral transmission was analyzed under different growing temperatures. The results show (i) the high efficiency of Movento® to protect rapeseed and sugar beet plants against the establishment of aphid colonies, (ii) the impact of temperature on the persistence of the Movento® aphicid properties and (iii) a decrease of approximately 10% of the viral transmission on treated plants. These observations suggest a beneficial effect of Movento® on the sanitary quality of treated crops by directly reducing primary infections and indirectly altering, through aphid mortality, secondary infections on which the spread of disease within field depends. These data constitute important elements for the future development of management strategies to protect crops against aphid-transmitted viruses.

Management of yellow dwarf disease in Europe in a post-neonicotinoid agriculture.

Barley/cereal yellow dwarf viruses (YDVs) cause yellow dwarf disease (YDD), which is a continuous risk to cereals production worldwide. These viruses cause leaf yellowing and stunting, resulting in yield reductions of up to 80%. YDVs have been a consistent but low-level problem in European cereal cultivation for the last three decades, mostly due to the availability of several effective insecticides (largely pyrethroids and more recently neonicotinoids) against aphid vectors. However, this has changed recently, with many insecticides being lost, culminating in a recent European Union (EU) regulation prohibiting outdoor use of the neonicotinoid-insecticide compounds. This change is coupled with the growing challenge of insecticide-resistant aphids, the lack of genetic resources against YDVs, and a knowledge deficit around the parameters responsible for the emergence and spread of YDD. This means that economic sustainability of cereal cultivation in several European countries including France and United Kingdom is now again threatened by this aphid-vectored viral disease. In this review, we summarize the current knowledge on the YDV pathosystem, describe management options against YDD, analyse the impacts of the neonicotinoid ban in Europe, and consider future strategies to control YDV. © 2020 Society of Chemical Industry.

Fluorescent-based techniques for viral detection, quantification, and characterization.

Fluorescent-based technologies offer opportunities for developing new assays for detection, quantification, and characterization of viral isolates. According to the intrinsic characteristics of fluorescent-based tools (high specificity, sensitivity, and reliability), such type of molecular assays makes possible investigations on original studies such as evolutionary processes (including fitness measurement of isolates), quantitative epidemiology, or the analysis of synergism and antagonism between closely related isolates. The development of these tools is very simple and requires, in complement to basic molecular knowledge such as extraction, cloning, and (RT)-PCR procedures, only the identification of short specific sequence(s) in the targeted viral genome. The Single Nucleotide Polymorphism (SNP) and the 'real-time' RT-PCR assays are proposed as fluorescent-based tools for qualitative and quantitative viral detection, respectively. Moreover, the SNaPshot technology is described as method for isolate characterization.

Analysis of population structures of viral isolates using single-strand conformation polymorphism method.

The analysis of viral populations requires the use of techniques that describe characteristics of individuals. The single-strand conformation polymorphism (SSCP) makes possible the identification of genetic differences between viral sequences and constitutes an alternative to the expensive and time-consuming cloning and sequencing strategies. Applied to small genomic regions (from 100 to 500 bases in length), SSCP patterns could describe, under appropriate experimental conditions, single nucleotide variations in the studied sequence. The different steps of a complete SSCP procedure, from sampling to pattern analysis (including nucleic acid extraction, RT-PCR amplification, double-stranded DNA quantification, polyacrylamide gel preparation, electrophoresis conditions, and staining procedures), are described using a region (500 bases) of the barley yellow dwarfvirus-PAV (BYDV-PAV, Luteovirus) genome as molecular target.

Functional Transcomplementation between Wheat Dwarf Virus Strains in Wheat and Barley.

Wheat dwarf virus, transmitted by the leafhopper Psammotettix alienus in a persistent, non-propagative manner, infects numerous species from the Poaceae family. Data associated with wheat dwarf virus (WDV) suggest that some isolates preferentially infect wheat while other preferentially infect barley. This allowed to define the wheat strain and the barley strain. There are contradictory results in the literature regarding the ability of each of these two strains to infect its non-preferred host. To improve knowledge on the interactions between WDV strains and barley and wheat, transmission experiments were carried out using barcoded P. alienus and an experimental design based on single/sequential acquisitions of WDV strains and on transmissions to wheat and barley. Results showed that (I) WDV strains are transmitted with similar efficiencies by P. alienus males, females and larvae, (II) WDV wheat and barley strains do not infect barley and wheat plants, respectively, and (III) a functional transcomplementation between the wheat and barley strains allows a mixed infection of barley and wheat. The described ability of each WDV strain to infect a non-host plant in the presence of the other viral strain must be considered to analyze data available on WDV host range.

A complex eIF4E locus impacts the durability of va resistance to Potato virus Y in tobacco.

Many recessive resistances against potyviruses are mediated by eukaryotic translation initiation factor 4E (eIF4E). In tobacco, the va resistance gene commonly used to control Potato virus Y (PVY) corresponds to a large deletion affecting the eIF4E-1 gene on chromosome 21. Here, we compared the resistance durability conferred by various types of mutations affecting eIF4E-1 (deletions of various sizes, frameshift or nonsense mutations). The 'large deletion' genotypes displayed the broadest and most durable resistance, whereas frameshift and nonsense mutants displayed a less durable resistance, with rapid and frequent apparition of resistance-breaking variants. In addition, genetic and transcriptomic analyses revealed that resistance durability is strongly impacted by a complex genetic locus on chromosome 14, which contains three other eIF4E genes. One of these, eIF4E-3, is rearranged as a hybrid gene between eIF4E-2 and eIF4E-3 (eIF4E-2-3 ) in the genotypes showing the most durable resistance, while eIF4E-2 is differentially expressed between the tested varieties. RNA-seq and quantitative reverse transcriptase-polymerase chain reaction experiments demonstrated that eIF4E-2 expression level is positively correlated with resistance durability. These results suggest that besides the nature of the mutation affecting eIF4E-1, three factors linked with a complex locus may potentially impact va durability: loss of an integral eIF4E-3, presence of eIF4E-2-3 and overexpression of eIF4E-2. This latter gene might act as a decoy in a non-productive virus-plant interaction, limiting the ability of PVY to evolve towards resistance breaking. Taken together, these results show that va resistance durability can in large part be explained by complex redundancy effects in the eIF4E gene family.

A multiple single nucleotide polymorphisms interrogation assay for reliable Potato virus Y group and variant characterization.

The complex Potato virus Y classification, including groups (PVYN and PVYO) and variants (PVYNTN and PVYN-W), is based mainly on biological properties of isolates. Published PVY detection tools targeting markers not associated with biological properties could fail to assign correctly isolates in the current classification. To improve PVY detection tools, a single nucleotide polymorphism (SNaPshot) detection assay was developed. The technique was adapted to target the T/C9259, A/C2271, G/C8573 and A/G2213 PVY polymorphic nucleotides. The "TAGA", "CCCG", "CACA" and "CAGA" four-digit codes associated with tested samples allowed identification of PVYN, PVYO, PVYN-W and PVYNTN isolates, respectively. The PVY SNaPshot procedure is efficient and reliable for PVY detection and characterization in samples containing as few as 10(2) viral RNA copies. Moreover, PVY group assignment is possible for fractions containing only 10 copies of a PVY RNA genome. Finally, the SNaPshot assay allows PVY(N)/PVYO dual characterization for mixed samples containing PVYN/PVYO quantity ratios in the range of 0.1-10. This innovative SNaPshot tool improved clearly PVY diagnostic assays described previously by targeting simultaneously major functional markers and sequence unlinked to biological properties used separately in PVY detection tools available currently.

Mating behavior of Psammotettix alienus (Hemiptera: Cicadellidae).

The Wheat dwarf virus, the causal agent of the wheat dwarf disease, is transmitted by leafhoppers from the genus Psammotettix and currently the main protection strategy is based on the use of insecticide treatments. Sustainable management strategies for insect vectors should include methods that are targeted to disrupt reproductive behavior and here we investigated the mating behavior of Psammotettix alineus (Dahlbom 1850) in order to determine the role of vibrational signals in intra-specific communication and pair formation. Both genders spontaneously emit species- and sex-specific calling songs that consisted of regularly repeated pulse trains and differ primarily in pulse train duration and pulse repetition time. Females preferred the conspecific male calling song. After a coordinated exchange of pulse trains, the male approached the stationary female. During the close range courtship and also immediately prior to copulatory attempts distinct male vibrational signals associated with wing flapping and wing vibrations were recorded from the substrate. In the presence of a receptive female, competing males emitted vibrational signals most likely aimed to interfere with male-female interaction. Mated females regained sexual receptivity after they laid eggs. Although results suggest that the viruliferous status of insects may have an effect on vibrational songs, our current results did not reveal a significant effect of virus on leafhopper performance in mating behavior. However, this study also suggests, that detailed understanding of plant-vector-virus interactions relevant for vector mating behavior is essential for trying new approaches in developing future control practices against plant viruses transmitted by insect vectors.

Using sensitivity analysis to identify key factors for the propagation of a plant epidemic.

Identifying the key factors underlying the spread of a disease is an essential but challenging prerequisite to design management strategies. To tackle this issue, we propose an approach based on sensitivity analyses of a spatiotemporal stochastic model simulating the spread of a plant epidemic. This work is motivated by the spread of sharka, caused by plum pox virus, in a real landscape. We first carried out a broad-range sensitivity analysis, ignoring any prior information on six epidemiological parameters, to assess their intrinsic influence on model behaviour. A second analysis benefited from the available knowledge on sharka epidemiology and was thus restricted to more realistic values. The broad-range analysis revealed that the mean duration of the latent period is the most influential parameter of the model, whereas the sharka-specific analysis uncovered the strong impact of the connectivity of the first infected orchard. In addition to demonstrating the interest of sensitivity analyses for a stochastic model, this study highlights the impact of variation ranges of target parameters on the outcome of a sensitivity analysis. With regard to sharka management, our results suggest that sharka surveillance may benefit from paying closer attention to highly connected patches whose infection could trigger serious epidemics.