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Olive quick decline syndrome (OQDS) is a disorder associated with bacterial infections caused by Xylella fastidiosa subsp. pauca ST53 in olive trees. Metabolic profile changes occurring in infected olive trees are still poorly investigated, but have the potential to unravel reliable biomarkers to be exploited for early diagnosis of infections. In this study, an untargeted metabolomic method using high-performance liquid chromatography coupled to quadrupole-time-of-flight high-resolution mass spectrometry (HPLC-ESI-Q-TOF-MS) was used to detect differences in samples (leaves) from healthy (Ctrl) and infected (Xf) olive trees. Both unsupervised and supervised data analysis clearly differentiated the groups. Different metabolites have been identified as potential specific biomarkers, and their characterization strongly suggests that metabolism of flavonoids and long-chain fatty acids is perturbed in Xf samples. In particular, a decrease in the defence capabilities of the host after Xf infection is proposed because of a significant dysregulation of some metabolites belonging to flavonoid family. Moreover, oleic acid is confirmed as a putative diffusible signal factor (DSF). This study provides new insights into the host-pathogen interactions and confirms LC-HRMS-based metabolomics as a powerful approach for disease-associated biomarkers discovery in plants. Graphical abstract

Xylella fastidiosa (Xf) is a harmful plant pathogenic bacterium, able to infect over 500 plant species worldwide. Successful eradication and containment strategies for harmful pathogens require large-scale monitoring techniques for the detection of infected hosts, even when they do not display visual symptoms. Although a previous study using airborne hyperspectral and thermal imagery has shown promising results for the early detection of Xf-infected olive (Olea europaea) trees, further work is needed when adopting these techniques for large scale monitoring using multispectral cameras on board airborne platforms and satellites. We used hyperspectral and thermal imagery collected during a two-year airborne campaign in a Xf-infected area in southern Italy to assess the performance of spectrally constrained machine-learning algorithms for this task. The algorithms were used to assess multispectral bandsets, selected from the original hyperspectral imagery, that were compatible with large-scale monitoring from unmanned platforms and manned aircraft. In addition, the contribution of solar–induced chlorophyll fluorescence (SIF) and the temperature-based Crop Water Stress Index (CWSI) retrieved from hyperspectral and thermal imaging, respectively, were evaluated to quantify their relative importance in the algorithms used to detect Xf infection. The detection performance using support vector machine algorithms decreased from ∼80% (kappa, κ = 0.42) when using the original full hyperspectral dataset including SIF and CWSI to ∼74% (κ = 0.36) when the optimal set of six spectral bands most sensitive to Xf infection were used in addition to the CWSI thermal indicator. When neither SIF nor CWSI were used, the detection yielded less than 70% accuracy (decreasing κ to very low performance, 0.29), revealing that tree temperature was more important than chlorophyll fluorescence for the Xf detection. This work demonstrates that large-scale Xf monitoring can be supported using airborne platforms carrying multispectral and thermal cameras with a limited number of spectral bands (e.g., six to 12 bands with 10 nm bandwidths) as long as they are carefully selected by their sensitivity to the Xf symptoms. More precisely, the blue (bands between 400 and 450 nm to derive the NPQI index) and thermal (to derive CWSI from tree temperature) were the most critical spectral regions for their sensitivity to Xf symptoms in olive., Data collection was partially supported by the European Union’s Horizon 2020 research and innovation program through grants to the POnTE (Pest Organisms threatening Europe; grant 635646 from European Union’s Horizon 2020 Framework Research Programme) and XF-ACTORS (Xylella fastidiosa Active Containment Through a Multidisciplinary-Oriented Research Strategy; grant 727987 from European Union’s Horizon 2020 Framework Research Programme) projects. A. Hornero was supported by a research fellowship DTC GEO 29 “Detection of global photosynthesis and forest health from space” from the Science Doctoral Training Centre (Swansea University, UK).

Since 2013,Xylella fastidiosaWells et al. has been reported to infect several hosts and to be present in different areas of Europe. The main damage has been inflicted on the olive orchards of southern Apulia (Italy), where a severe disease associated withX. fastidiosasubspeciespaucastrain De Donno has led to the death of millions of trees. This dramatic and continuously evolving situation has led to European and national (Italian and Spanish) measures being implemented to reduce the spread of the pathogen and the associated olive quick decline syndrome (OQDS). Research has been also carried out to find solutions to better and directly fight the bacterium and its main insect vector,Philaenus spumariusL. In the course of this frantic effort, several treatments based on chemical or biological substances have been tested, in addition to plant breeding techniques and integrated pest management approaches. This review aims to summarize the attempts made so far and describe the prospects for better management of this serious threat, which poses alarming questions for the future of olive cultivation in the Mediterranean basin and beyond.

This research was carried out in order to evaluate the presence and distribution of viral infections causing severe disease in eggplant plants collected from different districts in Bekaa valley, Lebanon. Most infected plants showed virus-like symptoms consisting predominantly of leaf blotch, mottling chlorotic and ring spots, leaf twisting and plant dwarf were also observed in the visited fields. Symptomatic and asymptomatic plants were collected and screened by ELISA test for the presence of several different pathogenic viruses potentially present in the area. Results showed that potato virus Y (PVY) was the most prevalent virus found by ELISA (detected in the 15.3% of the tested plants), followed by eggplant mottled dwarf virus (EMDV, 2.9%) and cucumber mosaic virus (CMV, 1.2%), while tomato spotted wilt virus (TSWV), alfalfa mosaic virus (AMV) and pepper mottle virus (PepMoV) were not detected. Biological indexing of symptomatic ELISA-negative plants, followed by electron microscopy, indicated the presence of virus-like particles of the genus Potexvirus, which was subsequently confirmed as potato virus X (PVX) by RT-PCR and Sanger sequencing. PVX was found in 35.3% of the tested plants, all sampled in the northern Bekaa area. In a phylogenetic analysis, the partial coat protein gene sequence of a selected Lebanese isolate, PVX-AK1, clustered together with other PVX isolates from Asia. Furthermore, the 124-aa sequence of PVX-AK1 shared 100% identity with PVX-UK3, an isolate which is known as avirulent in potato genotypes carrying either Nx or Rx resistance genes. This work revealed a picture of the previously uninvestigated phytosanitary status of eggplant crops in an important horticultural area of Lebanon.

In the last decade, the bacterial pathogen Xylella fastidiosa has devastated olive trees throughout Apulia region (Southern Italy) in the form of the disease called “Olive Quick Decline Syndrome” (OQDS). This study describes changes in the metabolic profile due to the infection by X. fastidiosa subsp. pauca ST53 in artificially inoculated young olive plants of the susceptible variety Cellina di Nardò. The test plants, grown in a thermo-conditioned greenhouse, were also co-inoculated with some xylem-inhabiting fungi known to largely occur in OQDS-affected trees, in order to partially reproduce field conditions in terms of biotic stress. The investigations were performed by combining NMR spectroscopy and MS spectrometry with a non-targeted approach for the analysis of leaf extracts. Statistical analysis revealed that Xylella-infected plants were characterized by higher amounts of malic acid, formic acid, mannitol, and sucrose than in Xylella-non-infected ones, whereas it revealed slightly lower amounts of oleuropein. Attention was paid to mannitol which may play a central role in sustaining the survival of the olive tree against bacterial infection. This study contributes to describe a set of metabolites playing a possible role as markers in the infections by X. fastidiosa in olive.

The introduction of the Xylella fastidiosa Wells bacterium into Apulia (South Italy) has caused the massive dieback of olive trees, and is threatening olive production throughout the Mediterranean Region. The key vector of X. fastidiosa in Europe is the spittlebug Philaenus spumarius L. The dispersal capabilities of P. spumarius are poorly known, despite being a key parameter for the prediction of the spread of the bacterium. In this study, we have examined the dispersal of P. spumarius adults in two different agroecosystems in Italy: an olive grove in Apulia (Southern Italy) and a meadow in Piedmont (Northern Italy). Insects were marked with albumin and released during seven independent trials over 2 yr. The recapture data were pooled separately for each agroecosystem and used to estimate the dispersal kernels of P. spumarius in the olive grove and in the meadow. The diffusion coefficient estimate for P. spumarius was higher in the meadow than in the olive grove. The median distance from the release point for 1 d of dispersal was 26 m in the olive grove and 35 m in the meadow. On the basis of our model, we estimated that 50% of the spittlebug population remained within 200 m (98% within 400 m) during the 2 mo period of high abundance of the vector on olives in Apulia. The dispersal of P. spumarius is thus limited to some hundreds of meters throughout the whole year, although it can be influenced to a great extent by the structure of the agroecosystem.

In 2013, Xylella fastidiosa spp. pauca was first reported in Puglia, Italy, causing the olive quick decline syndrome (OQDS). Since then the disease has spread, prompting the initiation of management measures to contain the outbreak. Estimates of the shape of the disease front and the rate of area expansion are needed to inform management, e.g. the delineation of buffer zones. However, empirical estimates of the invasion front and the rate of spread of OQDS are not available. Here, we analysed the hundreds of thousands of records of monitoring data on disease occurrence in Puglia to estimate the shape of the invasion front and the rate of movement of the front. The robustness of estimation was checked using simulation. The shape of the front was best fitted by a logistic function while using a beta-binomial error distribution to model variability around the expected proportion of infected trees. The estimated rate of movement of the front was 10.0 km per year (95% confidence interval: 7.5–12.5 km per year). This rate of movement is at the upper limit of previous expert judgements. The shape of the front was flatter than expected. The fitted model indicates that the disease spread started approximately in 2008. This analysis underpins projections of further disease spread and the need for preparedness in areas that are still disease free.

The plant-pathogenic bacteriumXylella fastidiosawas first reported in Europe in 2013, in the province of Lecce, Italy, where extensive areas were affected by the olive quick decline syndrome, caused by the subsp.pauca. In Alicante, Spain, almond leaf scorch, caused byX. fastidiosasubsp.multiplex, was detected in 2017. The effects of climatic and spatial factors on the geographic distribution ofX. fastidiosain these two infested regions in Europe were studied. The presence/absence data ofX. fastidiosain the official surveys were analyzed using Bayesian hierarchical models through the integrated nested Laplace approximation (INLA) methodology. Climatic covariates were obtained from the WorldClim v.2 database. A categorical variable was also included according to Purcell’s minimum winter temperature thresholds for the risk of occurrence of Pierce’s disease of grapevine, caused byX. fastidiosasubsp.fastidiosa. In Alicante, data were presented aggregated on a 1km grid (lattice data), where the spatial effect was included in the model through a conditional autoregressive structure. In Lecce, data were observed at continuous locations occurring within a defined spatial domain (geostatistical data). Therefore, the spatial effect was includedviathe stochastic partial differential equation approach. In Alicante, the pathogen was detected in all four of Purcell’s categories, illustrating the environmental plasticity of the subsp.multiplex. Here, none of the climatic covariates were retained in the selected model. Only two of Purcell’s categories were represented in Lecce. The mean diurnal range (bio2) and the mean temperature of the wettest quarter (bio8) were retained in the selected model, with a negative relationship with the presence of the pathogen. However, this may be due to the heterogeneous sampling distribution having a confounding effect with the climatic covariates. In both regions, the spatial structure had a strong influence on the models, but not the climatic covariates. Therefore, pathogen distribution was largely defined by the spatial relationship between geographic locations. This substantial contribution of the spatial effect in the models might indicate that the current extent ofX. fastidiosain the study regions had arisen from a single focus or from several foci, which have been coalesced.

Plant pathogens cause significant losses to agricultural yields and increasingly threaten food security1, ecosystem integrity and societies in general2–5. Xylella fastidiosa is one of the most dangerous plant bacteria worldwide, causing several diseases with profound impacts on agriculture and the environment6. Primarily occurring in the Americas, its recent discovery in Asia and Europe demonstrates that X. fastidiosa’s geographic range has broadened considerably, positioning it as a reemerging global threat that has caused socioeconomic and cultural damage7,8. X. fastidiosa can infect more than 350 plant species worldwide9, and early detection is critical for its eradication8. In this article, we show that changes in plant functional traits retrieved from airborne imaging spectroscopy and thermography can reveal X. fastidiosa infection in olive trees before symptoms are visible. We obtained accuracies of disease detection, confirmed by quantitative polymerase chain reaction, exceeding 80% when high-resolution fluorescence quantified by three-dimensional simulations and thermal stress indicators were coupled with photosynthetic traits sensitive to rapid pigment dynamics and degradation. Moreover, we found that the visually asymptomatic trees originally scored as affected by spectral plant-trait alterations, developed X. fastidiosa symptoms at almost double the rate of the asymptomatic trees classified as not affected by remote sensing. We demonstrate that spectral plant-trait alterations caused by X. fastidiosa infection are detectable previsually at the landscape scale, a critical requirement to help eradicate some of the most devastating plant diseases worldwide.

Xylella fastidiosa Wells et al. 1987 (family Xanthomonadaceae) is a Gram-negative, non-motile, non-sporing bacterium endowed with a very peculiar biological and epidemiological behaviour: it colonizes the xylem vessels of the hosts and is transmitted from plant to plant by xylem fluid-feeding insects (leafhoppers of the family Cicadellidae). Because of its polyphagy, when X. fastidiosa enters a new environment with congenial climatic conditions, it becomes rapidly entrenched in the territory. This is what has presumably taken place in Salento, an area where, a few years ago, a destructive disease of olive (Olea europaea L.) developed, denoted olive quick decline syndrome (OQDS), together with a leaf scorch condition of oleander, cherry and almond. Information stemming from studies carried out by the Bari team has laid the basis for implementing a strategy for OQDS containment, based on: (i) accurate surveillance of the territory north of the infected area; (ii) elimination of inoculum sources in newly developed infection foci through the uprooting of infected hosts and the surrounding healthy plants; and (iii) control of vectors by mechanical weeding in late winter/spring to kill juvenile forms, followed by insecticide treatments against the adults when, in late spring, they move from weeds to olives.

Citrus tristeza virus (CTV) is the pathogen causing tristeza diseases in several Rutaceae species and leading to significant economic damage to citrus worldwide. The Ctv locus provides broad spectrum resistance to CTV in Poncirus trifoliata L. Raf. This locus is present also in tolerant and susceptible species, so an epigenetic mechanism of Ctv expression regulation was proposed. Indeed, a difference in plant 24-nt sRNAs distribution corresponding to CTV.20 gene was previously observed in susceptible species following CTV infection. This gene, encoding for a plant virus movement-like protein, was investigated as a candidate gene for CTV susceptibility. Here, we show the presence of differences in methylation status of a specific region of CTV.20 in two susceptible species, sour and sweet orange, following CTV infection. On the contrary, no significant differences were observed in the tolerant Citrange carrizo following the infection. Moreover, a hypermethylation of the whole CTV.20 gene was observed in Citrange carrizo both healthy and infected, and in healthy sour and sweet orange. This preliminary study allows hypothesizing a possible role of methylation in regulation of CTV.20 expression involved in the CTV susceptibility.

Pathogen introductions have led to numerous disease outbreaks in naive regions of the globe. The plant pathogen Xylella fastidiosa has been associated with various recent epidemics in Europe affecting agricultural crops, such as almond, grapevine, and olive, but also endemic species occurring in natural forest landscapes and ornamental plants. We compared whole-genome sequences of X. fastidiosa subspecies multiplex from America and strains associated with recent outbreaks in southern Europe to infer their likely origins and paths of introduction within and between the two continents. Phylogenetic analyses indicated multiple introductions of X. fastidiosa subspecies multiplex into Italy, Spain, and France, most of which emerged from a clade with limited genetic diversity with a likely origin in California, USA. The limited genetic diversity observed in X. fastidiosa subspecies multiplex strains originating from California is likely due to the clade itself being an introduction from X. fastidiosa subspecies multiplex populations in the southeastern United States, where this subspecies is most likely endemic. Despite the genetic diversity found in some areas in Europe, there was no clear evidence of recombination occurring among introduced X. fastidiosa strains in Europe. Sequence type taxonomy, based on multilocus sequence typing (MLST), was shown, at least in one case, to not lead to monophyletic clades of this pathogen; whole-genome sequence data were more informative in resolving the history of introductions than MLST data. Although additional data are necessary to carefully tease out the paths of these recent dispersal events, our results indicate that whole-genome sequence data should be considered when developing management strategies for X. fastidiosa outbreaks., This work was funded by the following awards: XF-ACTORS (Xylella fastidiosa Active Containment Through a Multidisciplinary-Oriented Research Strategy, grant 727987 from the European Union’s Horizon 2020 Framework Research Programme), E-RTA2017-00004-C02 (Desarrollo de estrategias de erradicación, contención y control de X. fastidiosa en España from “Programa Estatal de I+D+I Orientada a los Retos de la Sociedad of the Spanish Government” and FEDER), the Spanish Olive Oil Interprofessional, and the California Department of Food and Agriculture Pierce’ Disease Research Program.

Olive quick decline syndrome (OQDS) is a devastating disease of olive trees in the Salento region, Italy. This disease is caused by the bacterium Xylella fastidiosa, which is widespread in the outbreak area, however, the &ldquo, Leccino&rdquo, variety of olives has proven to be resistant with fewer symptoms and lower bacterial populations than the &ldquo, Ogliarola salentina&rdquo, variety. We completed an empirical study to determine the mineral and trace element contents (viz, ionome) of leaves from infected trees comparing the two varieties, to develop hypotheses related to the resistance of Leccino trees to X. fastidiosa infection. All samples from both cultivars tested were infected by X. fastidiosa, even if leaves were asymptomatic at the time of collection, due to the high disease pressure in the outbreak area and the long incubation period of this disease. Leaves were binned for the analysis by variety, field location, and infected symptomatic and infected asymptomatic status by visual inspection. The ionome of leaf samples was determined using inductively coupled plasma optical emission spectroscopy (ICP-OES) and compared with each other. These analyses showed that Leccino variety consistently contained higher manganese (Mn) levels compared with Ogliarola salentina, and these levels were higher in both infected asymptomatic and infected symptomatic leaves. Infected asymptomatic and infected symptomatic leaves within a host genotype also showed differences in the ionome, particularly a higher concentration of calcium (Ca) and Mn levels in the Leccino cultivar, and sodium (Na) in both varieties. We hypothesize that the ionome differences in the two varieties contribute to protection against disease caused by X. fastidiosa infection.

Diseases caused by Xylella fastidiosa are among the most destructive for several agricultural productions. A deadly disease of olive, termed olive quick decline syndrome, is one of the most recent examples of the severe impacts caused by the introduction and spread of this bacterium in new ecosystems with favorable epidemiological conditions. Deciphering the cascade of events leading to the development of severe alterations in the susceptible host plants is a priority of several research programs investigating strategies to mitigate the detrimental impacts of the infections. However, in the case of olives, the long latent period (&gt, 1 year) makes this pathosystem not amenable for such studies. We have inoculated alfalfa (Medicago sativa) with the olive-infecting strain &ldquo, De Donno&rdquo, isolated from a symptomatic olive in Apulia (Italy), and we demonstrated that this highly pathogenic strain causes an overactive reaction that ends up with the necrosis of the inoculated stem, a reaction that differs from the notoriously Alfalfa Dwarf disease, caused by X. fastidiosa strains isolated from grapes and almonds. RNASeq analysis showed that major plant immunity pathways are activated, in particular, several calcium transmembrane transporters and enzymes responsible for the production of reactive oxygen species (ROS). Signs of the necrotic reaction are anticipated by the upregulation of genes responsible for plant cell death and the hypersensitive reaction. Overall the whole infection process takes four months in alfalfa, which makes this pathosystem suitable for studies involving either the plant response to the infection or the role of Xylella genes in the expression of symptoms.

An outbreak of Xylella fastidiosa subsp. multiplex sequence type ST6 was discovered in 2017 in mainland Spain affecting almond trees. Two cultured almond strains, “ESVL” and “IVIA5901,” were subjected to high throughput sequencing and the draft genomes assembled. Phylogenetic analysis conclusively indicated they belong to the subspecies multiplex, and pairwise comparisons of the chromosomal genomes showed an average nucleotide identity higher than 99%. Interestingly, the two strains differ for the presence of the plasmids pXF64-Hb_ESVL and pUCLA-ESVL detected only in the ESVL strain. The availability of these draft genomes contribute to extend the European genomic sequence dataset, a first step toward setting new research to elucidate the pathway of introduction and spread of the numerous strains of this subspecies so far detected in Europe., This work was funded by European Union’s Horizon 2020 Framework Research Programme Projects XF-ACTORS (Xylella fastidiosa Active Containment Through a Multidisciplinary-Oriented Research Strategy grant 727987) and MSCA-RISE-2016 CURE-XF (Capacity Building and Raising Awareness in Europe and in Third Countries to Cope with Xylella fastidiosa); COST Action CA16107 EuroXanth, supported by European Cooperation in Science and Technology; and Project Desarrollo de estrategias de erradicación, contención y control de en España: Diagnóstico, estructura genética y gama de huéspedes project E-RTA2017-00004-C06-02 from Programa Estatal de I+D+I Orientada a los Retos de la Sociedad of the Spanish Government. This work used the Vincent J. Coates Genomics Sequencing Laboratory at University of California, Berkeley, supported by NIH S10 OD018174 Instrumentation Grant.

Resistance-breaking (RB) strains constitute a clade of biological and genetically distinct isolates of Citrus tristeza virus (CTV) that replicate and move systemically in Poncirus trifoliata (trifoliate orange), resistant to other known strains of CTV. Molecular markers have been developed by comparative genome analysis to allow quick identification of potential RB isolates. Here, methods are described to identify and characterize RB strains by reverse transcription-polymerase chain reaction (RT-PCR), quantitative real-time RT-PCR (RT-qPCR), full-length genome sequencing, and biological indexing.

Diseases associated with Xylella fastidiosa have been described mostly in North and South America. However, during the last five years, widespread X. fastidiosa infections have been reported in a constrained area of the Apulia region (southern Italy), in olives trees suffering a severe disease, denoted as Olive Quick Decline Syndrome (OQDS). Because many xylem sap-feeding insects can function as vectors for the transmission of this exotic pathogen in EU, several research programs are ongoing to assess the role of candidate vectors in the spread of the infections. Initial investigations identified Philaenus spumarius (L.) as the predominant vector species in the olive orchards affected by the OQDS. Additional experiments have been carried out during 2016 and 2017 to assess the role of other species. More specifically, adults of the spittlebugs Philaenus italosignus Drosopolous and Remane, Neophilaenus campestris (Fallen) and of the planthopper Latilica tunetana (Matsumura) (Issidae) have been tested in transmission experiments to assess their ability to acquire the bacterium from infected olives and to infect different susceptible hosts (olives, almond, myrtle &ndash, leaf milkwort, periwinkle). Acquisition rates determined by testing individual insects in quantitative PCR assays, ranging from 5.6% in N. campestris to 22.2% in P. italosignus, whereas no acquisition was recorded for L. tunetana. Successful transmissions were detected in the recipient plants exposed to P. italosignus and N. campestris, whereas no trasmissions occurred with L. tunetana. The known vector Philaenus spumarius has been included in all the experiments for validation. The systematic surveys conducted in 2016 and 2017 provided further evidence on the population dynamics and seasonal abundance of the spittlebug populations in the olive groves.

The vector-borne bacterial pathogenXylella fastidiosais widely distributed in the Americas; in the last decade it has emerged as a serious threat for agricultural crops, natural environment and landscape in Europe. Following the first EU outbreak in 2013 in southern Italy, associated with a severe disease in olive trees, annual mandatory surveys are now in place in the Member States, leading to the discovery of bacterial outbreaks in different countries. Among the latest findings, an outbreak has been reported in the Italian region of Tuscany, with infections identified in seven different plant species. In this work, we report the isolation and the genetic characterization of isolates associated with this newly discovered outbreak. Multilocus sequence typing approach revealed the occurrence of isolates harbouring a new sequence type, denoted ST87, genetically related to strains of subsp.multiplex, but different from the genotypes of this subspecies previously characterized in Europe. Five cultured strains were successfully recovered from four of the seven host plants, an important achievement for advancing the studies on genomics and pathogenicity of theseisolates and thus assess their potential threat for European agriculture.

We report the complete annotated genome sequence of the plant-pathogenic bacterium Xylella fastidiosa subsp. fastidiosa strain IVIA5235.

Following a request from the European Commission, the EFSA Plant Health Panel updated its pest categorisation of Xylella fastidiosa, previously delivered as part of the pest risk assessment published in 2015. X. fastidiosa is a Gram‐negative bacterium, responsible for various plant diseases, including Pierce's disease, phony peach disease, citrus variegated chlorosis, olive quick decline syndrome, almond leaf scorch and various other leaf scorch diseases. The pathogen is endemic in the Americas and is present in Iran. In the EU, it is reported in southern Apulia in Italy, on the island of Corsica and in the Provence‐Alpes‐Côte d'Azur region in France, as well as in the Autonomous region of Madrid, the province of Alicante and the Balearic Islands in Spain. The reported status is ‘transient, under eradication’, except for the Balearic Islands, Corsica and southern of Apulia, where the status is ‘present with a restricted distribution, under containment’. The pathogen is regulated under Council Directive 2000/29/EC and through emergency measures under http://eur-lex.europa.eu/legal-content/EN/TXT/?uri=CELEX:32015D0789 (as amended http://eur-lex.europa.eu/legal-content/EN/TXT/?uri=CELEX:32017D2352). The pest could enter the EU via host plants for planting and via infectious insect vectors. The host range includes hundreds of host species listed in the EFSA host plant database. In the EU, host plants are widely distributed and climatic conditions are favourable for its establishment. X. fastidiosa can spread by movement of host plants for planting and infectious insect vectors. X. fastidiosa is known to cause severe direct damage to major crops including almonds, citrus, grapevines, olives, stone fruits and also forest trees, landscape and ornamental trees, with high impacts. The criteria assessed by the Panel for consideration as a potential Union quarantine pest are met (the pathogen is present in the EU, but it has a restricted distribution and is under official control). X. fastidiosa is not considered as a regulated non‐quarantine pest (RNQP) as the pathogen may spread also via insect vector transmission.

Philaenus spumariusL. has been identified as the main vector ofXylella fastidiosa,the bacterium causing a disease denoted as Olive Quick Decline Syndrome (OQDS) in Southern Italy.From late 2017 to early 2018, only two commercial formulations of acetamiprid and delthametrin were officially registered in Italy. Hence, an urgent need exists to evaluate field efficacy of chemicals with different active ingredients, formulations, and modes of action to control the meadow spittlebug. From 2015 to 2017, six different field trials (A, B, C, D, E, and F) were conducted using a randomized block with six replicates per treatment including the untreated check and neonicotinoids as reference products with known efficacy against other Auchenorrhyncha. The efficacy and persistence of each application was determined by scoring spittlebug mortality at 3, 7, 10, and 15 DAT for the trials conducted in 2015, and up to 20 and 25 DAT in trials conducted in 2016 and 2017.

The olive quick decline syndrome (OQDS) is a disease that appeared suddenly a few years ago in the province of Lecce (Salento peninsula, southeastern Italy). Among the factors that may be involved in its aetiology, the most relevant is Xylella fastidiosa, a quarantine pathogen of American origin, whose presence in Italy represents its first confirmed record in the European Union. X. fastidiosa is a Gram-negative bacterium that invades the xylem of a wide range of hosts, from which it is acquired by xylem-feeding insect vectors and transferred to other plants. The bacterium multiplies within the plant vessels and occludes them, thus impairing water uptake. Besides olive, the Salentian strain of X. fastidiosa infects in nature a number of woody (almond, cherry) and shrubby (oleander, broom, Acacia saligna, Polygala myrtifolia, Westringia fruticosa, Rosmarinus officinalis, Rhamnus elaternus, Myrtus communis) hosts, with no evidence for grapevines or citrus being hosts. The bacterium was isolated in culture and identified as a genotype of X. fastidiosa subsp. pauca, molecularly identical to an isolate from Costa Rica. Philaenus spumarius (meadow spittlebug), a froghopper quite common in the Salento area where it thrives on olive, was identified as the main vector. Disease eradication and sanitation of infected olives are unfeasible. However, strategies are being enacted for restraining the spread of pathogen and vector(s) within the boundaries of the currently infected zone.

The dynamics of Xylella fastidiosa infections in the context of the endophytic microbiome was studied in field-grown plants of the susceptible and resistant olive cultivars Kalamata and FS17. Whole metagenome shotgun sequencing (WMSS) coupled with 16S/ITS rRNA gene sequencing was carried out on the same trees at two different stages of the infections: In Spring 2017 when plants were almost symptomless and in Autumn 2018 when the trees of the susceptible cultivar clearly showed desiccations. The progression of the infections detected in both cultivars clearly unraveled that Xylella tends to occupy the whole ecological niche and suppresses the diversity of the endophytic microbiome. However, this trend was mitigated in the resistant cultivar FS17, harboring lower population sizes and therefore lower Xylella average abundance ratio over total bacteria, and a higher α-diversity. Host cultivar had a negligible effect on the community composition and no clear associations of a single taxon or microbial consortia with the resistance cultivar were found with both sequencing approaches, suggesting that the mechanisms of resistance likely reside on factors that are independent of the microbiome structure. Overall, Proteobacteria, Actinobacteria, Firmicutes, and Bacteriodetes dominated the bacterial microbiome while Ascomycota and Basidiomycota those of Fungi., This research was funded by the EU H2020 Research Project XF-ACTORS "Xylella Fastidiosa Active Containment Through a multidisciplinary-Oriented Research Strategy" (Grant Agreement 727987) and by the Mountain Research Center-CIMO (UIDB/00690/2020) and BioISI (UIDB/04046/2020). The Funding Agencies are not responsible for any use that may be made of the information it contains.

© 2016, Koninklijke Nederlandse Planteziektenkundige Vereniging. After the first confirmed outbreak of Xylella fastidiosa in the European Union (EU), associated with an olive disease denoted olive quick decline syndrome, mandatory surveys are now carried out in the member States and inspections increased at EU entry points such as ports. Such activities led to the interception of X. fastidiosa-infected coffee plants in consignments originating from Central America. Similarly, the geographic expansion of the olive decline epidemic area of the Apulia region (southern Italy) prompted investigations to identify new host plants. Here we report the interception of three novel bacterial sequence types in Italy, based on multi-locus sequence typing, that cluster with different X. fastidiosa subspecies, illustrating the risk of the introduction of additional pathogen genetic diversity into Europe. In the epidemic area of Apulia, new foci as well as host plant species positive with X. fastidiosa, including cherry, myrtleleaf and rosemary, were found to be all infected with the same sequence type of this bacterium (ST53, or CoDiRO strain). This work highlights the limited knowledge of X. fastidiosa phylogenetic and phenotypic diversity, the risk of novel X. fastidiosa introductions via contaminated plant material, and corroborates other studies indicating that the Apulia epidemic emerged from a single introduction of this pathogen into the region.

In olive, several biological processes, including those related to drupe maturation and oil production, are adversely affected by biotic stress. Pesticides are an important, valuable input in modern oliviculture, still central to secure yield and safeguard olive oil quality. However, concerns over the effects of plant protection products on the environment, non-target organisms, and human health prompt the development and implementation of more integrated control strategies. Functional genomics for biotic stress tolerance is a promising area that needs to be explored to increase olive tolerance and reduce reliance on pesticides. A number of studies have recently described, on a genome-wide scale, the participation of genes in olive response to different biotic stresses. Moreover, genes involved in stress tolerance and related signaling networks have been also identified. This chapter presents recent advances in olive molecular response to its major biotic stresses (insects, fungi, bacteria, and viruses). This topic is presented in a larger context that includes the main biological features of the major olive biotic stressors.

© 2016, The Author(s). The recent introduction of Xylella fastidiosa in Europe and its involvement in the Olive Quick Decline Syndrome (OQDS) in Apulia (Salento, Lecce district, South Italy) led us to investigate the biology and transmission ability of the meadow spittlebug, Philaenus spumarius, which was recently demonstrated to transmit X. fastidiosa to periwinkle plants. Four xylem-sap-feeding insect species were found within and bordering olive orchards across Salento during a survey carried out from October 2013 to December 2014: P. spumarius was the most abundant species on non-olive vegetation in olive orchards as well as on olive foliage and was the only species that consistently tested positive for the presence of X. fastidiosa using real-time PCR. P. spumarius, whose nymphs develop within spittle on weeds during the spring, are likely to move from weeds beneath olive trees to olive canopy during the dry period (May to October 2014). The first X. fastidiosa-infective P. spumarius were collected in May from olive canopy: all the individuals previously collected on weeds tested negative for the bacterium. Experiments demonstrated that P. spumarius transmitted X. fastidiosa from infected to uninfected olive plants. Moreover, P. spumarius acquired X. fastidiosa from several host plant species in the field, with the highest acquisition rate from olive, polygala and acacia. Scanning electron microscopy (SEM) revealed bacterial cells resembling X. fastidiosa in the foreguts of adult P. spumarius. The data presented here are essential to plan an effective IPM strategy and limit further spread of the fastidious bacterium.

A dramatic outbreak of Xylella fastidiosa decimating olive was discovered in 2013 in Apulia, Southern Italy. This pathogen is a quarantine bacterium in the European Union (EU) and created unprecedented turmoil for the local economy and posed critical challenges for its management. With the new emerging threat to susceptible crops in the EU, efforts were devoted to gain basic knowledge on the pathogen biology, host, and environmental interactions (e.g., bacterial strain(s) and pathogenicity, hosts, vector(s), and fundamental drivers of its epidemics) in order to find means to control or mitigate the impacts of the infections. Field surveys, greenhouse tests, and laboratory analyses proved that a single bacterial introduction occurred in the area, with a single genotype, belonging to the subspecies pauca, associated with the epidemic. Infections caused by isolates of this genotype turned to be extremely aggressive on the local olive cultivars, causing a new disease termed olive quick decline syndrome. Due to the initial extension of the foci and the rapid spread of the infections, eradication measures (i.e., pathogen elimination from the area) were soon replaced by containment measures including intense border surveys of the contaminated area, removal of infected trees, and mandatory vector control. However, implementation of containment measures encountered serious difficulties, including public reluctance to accept control measures, poor stakeholder cooperation, misinformation from some media outlets, and lack of robust responses by some governmental authorities. This scenario delayed and limited containment efforts and allowed the bacterium to continue its rapid dissemination over more areas in the region, as shown by the continuous expansion of the official borders of the infected area. At the research level, the European Commission and regional authorities are now supporting several programs aimed to find effective methods to mitigate and contain the impact of X. fastidiosa on olives, the predominant host affected in this epidemic. Preliminary evidence of the presence of resistance in some olive cultivars represents a promising approach currently under investigation for long-term management strategies. The present review describes the current status of the epidemic and major research achievements since 2013.

Here, we provide the first insights into the transmission dynamics of the bacterium Xylella fastidiosa by the meadow spittlebug Philaenus spumarius, gathered through DC EPG (electrical penetration graph)-assisted transmission tests and comparative observations of the probing and feeding behavior of infective versus non-infective vectors on healthy olive plants. Bacterial cells binding to P. spumarius’ foregut occurred at a very low rate and in a time as short as 15 min spent by the insect in xylem ingestion or activities interspersed with xylem ingestion (interruption during xylem ingestion and resting). P. spumarius inoculation of bacterial cells into the xylem was exclusively associated with an early (ca. 2 to 7 min after the onset of the first probe) and occasional behavior, provisionally termed waveform Xe, presumably related to egestion regulated by pre-cibarial valve fluttering. Infective spittlebugs compared to non-infective ones exhibited: (i) longer non-probing and shorter xylem ingestion; (ii) longer duration of single non-probing events; (iii) fewer sustained ingestions (ingestion longer than 10 min) and interruptions of xylem activity (N); and (iv) longer time required to perform the first probe. These observations suggest difficulties in feeding of infective P. spumarius probably caused by the presence of X. fastidiosa within the foregut. Overall, our data indicate that likely short time—few minutes—is required for X. fastidiosa transmission by P. spumarius; thus, vector control strategies should aim at preventing spittlebug access to the host plant. Furthermore, our findings represent an important contribution for further research on the disruption of spittlebug–bacterium interactions.

In autumn 2013, the presence of Xylella fastidiosa, a xylem-limited Gram-negative bacterium, was detected in olive stands of an area of the Ionian coast of the Salento peninsula (Apulia, southern Italy), that were severely affected by a disease denoted olive quick decline syndrome (OQDS). Studies were carried out for determining the involvement of this bacterium in the genesis of OQDS and of the leaf scorching shown by a number of naturally infected plants other than olive. Isolation in axenic culture was attempted and assays were carried out for determining its pathogenicity to olive, oleander and myrtle-leaf milkwort. The bacterium was readily detected by quantitative polymerase chain reaction (qPCR) in all diseased olive trees sampled in different and geographically separated infection foci, and culturing of 51 isolates, each from a distinct OQDS focus, was accomplished. Needle-inoculation experiments under different environmental conditions proved that the Salentinian isolate De Donno belonging to the subspecies pauca is able to multiply and systemically invade artificially inoculated hosts, reproducing symptoms observed in the field. Bacterial colonization occurred in prick-inoculated olives of all tested cultivars. However, the severity of and timing of symptoms appearance differed with the cultivar, confirming their differential reaction.

Background The recent Xylella fastidiosa subsp. pauca (Xfp) outbreak in olive (Olea europaea) groves in southern Italy is causing a destructive disease denoted Olive Quick Decline Syndrome (OQDS). Field observations disclosed that Xfp-infected plants of cv. Leccino show much milder symptoms, than the more widely grown and highly susceptible cv. Ogliarola salentina. To determine whether these field observations underlie a tolerant condition of cv. Leccino, which could be exploited for lessening the economic impact of the disease on the local olive industry, transcriptional changes occurring in plants of the two cultivars affected by Xfp were investigated. Results A global quantitative transcriptome profiling comparing susceptible (Ogliarola salentina) and tolerant (Leccino) olive cultivars, infected or not by Xfp, was done on messenger RNA (mRNAs) extracted from xylem tissues. The study revealed that 659 and 447 genes were differentially regulated in cvs Leccino and Ogliarola upon Xfp infection, respectively, whereas 512 genes were altered when the transcriptome of both infected cultivars was compared. Analysis of these differentially expressed genes (DEGs) shows that the presence of Xfp is perceived by the plants of both cultivars, in which it triggers a differential response strongly involving the cell wall. Up-regulation of genes encoding receptor-like kinases (RLK) and receptor-like proteins (RLP) is the predominant response of cv. Leccino, which is missing in cv. Ogliarola salentina. Moreover, both cultivars react with a strong re-modelling of cell wall proteins. These data suggest that Xfp elicits a different transcriptome response in the two cultivars, which determines a lower pathogen concentration in cv. Leccino and indicates that this cultivar may harbor genetic constituents and/or regulatory elements which counteract Xfp infection. Conclusions Collectively these findings suggest that cv. Leccino is endowed with an intrinsic tolerance to Xfp, which makes it eligible for further studies aiming at investigating molecular basis and pathways modulating its different defense response. Electronic supplementary material The online version of this article (doi:10.1186/s12864-016-2833-9) contains supplementary material, which is available to authorized users.