Xylella Research Articles

Diversity and composition of the bacterial communities associated with the Australian spittlebugs Bathyllus albicinctus and Philagra parva (Hemiptera: Aphrophoridae).

Spittlebugs and froghoppers (Hemiptera: Cercopoidea) are insects feeding on xylem, which potentially can cause significant economic damage worldwide by transmitting plant pathogenic bacteria such as Xylella fastidiosa. Australia and New Zealand are currently free from X. fastidiosa, but they are home to at least 45 native spittlebug species. Among these, the Australian natives Bathyllus albicinctus (Erichson, 1842) and Philagra parva (Donovan, 1805) are particularly widespread and can be found across southern and eastern Australia, with B. albicinctus also in New Zealand. The potential that both species might be capable of vectoring Xylella fastidiosa poses a substantial biosecurity risk if the bacterium were to invade these regions. In this study, we examined 87 spittlebug nymphs collected across 12 different host plant species, in five locations in Victoria, Australia. Our objective was to explore the factors influencing bacterial communities within and between these widespread spittlebug species, considering geographic location, insect phylogenetics, and host plant associations. We employed COI barcoding to assess insect genetic variation and 16S high throughput sequencing (HTS) metabarcoding to analyse bacterial microbiome diversity across various host plants. Our findings revealed minimal genetic divergence among spittlebug individuals in the same species, highlighting conspecificity despite conspicuous morphological divergences. On the other hand, we recorded significant variation in bacterial communities harboured by Bathyllus albicinctus nymphs feeding on different plants, even when these were collected within close proximity to each other. Therefore, host plant association appeared to shape the bacterial communities of spittlebugs more than insect genetic divergence or geographical location. These diverse bacterial communities could potentially facilitate transmission of plant pathogenic bacteria, underscoring the risk of widespread transmission among numerous plant hosts through insect-plant interactions. This study emphasizes the critical need to understand these complex interactions, particularly in the context of biosecurity.

Seasonal Dynamics and Distribution of Xylella fastidiosa in Infected Almond Trees.

This research focused on studying the dynamics of the bacterial pathogen Xylella fastidiosa in almond trees across different developmental stages. The objective was to understand the seasonal distribution and concentration of X. fastidiosa within almond trees. Different tree organs, including leaves, shoots, branches, fruits, flowers, and roots, from 10 X. fastidiosa-infected almond trees were sampled over 2 years. The incidence and concentration of X. fastidiosa were determined using qPCR and isolation. Throughout the study, X. fastidiosa was consistently absent from fruits, flowers, and roots, whereas it was detected in leaves as well as in shoots and branches. We demonstrate that the absence of X. fastidiosa in the roots is likely linked to the inability of this isolate to infect the peach-almond hybrid rootstock GF677. X. fastidiosa incidence in shoots and branches remained consistent throughout the year, whereas in leaf petioles, it varied across developmental stages, with lower detection during the early and late stages of the season. Similarly, viable X. fastidiosa cells were isolated from shoots and branches at all developmental stages, but no successful isolations were achieved from leaf petioles during the vegetative and nut growth stage. Studying the progression of almond leaf scorch symptoms in trees with initial infections showed that once symptoms emerged on one branch, symptomless branches were likely already infected by the bacterium. Therefore, selectively pruning symptomatic branches is unlikely to cure the tree. This study enhances our understanding of X. fastidiosa dynamics in almond trees and may have practical applications for its detection and control.

A Study of the Toxic Effect of Plant Extracts against Philaenus spumarius (Hemiptera: Aphrophoridae).

The meadow spittlebug Philaenus spumarius (Hemiptera: Aphrophoridae) is distributed in several habitats worldwide and has been recently recognized as the main vector of Xylella fastidiosa subsp. pauca. This bacterium has been associated with olive quick decline syndrome (OQDS) in the Salento Peninsula (Italy) and is responsible for extensive desiccation and die-off of olive trees. Current OQDS management strategies include the control of P. spumarius populations, mainly through the removal of weed hosts and insecticide treatments. In addition to the mandatory phytosanitary measures, the demand for new strategies compatible with integrated pest management is increasing. In this study, laboratory biological assays were performed to assess the potential toxic effect of vegetal formulations against P. spumarius adults. Two formulations were tested at different concentrations: Form A, an emulsion of 10% hot pepper-infused oil (Capsicum annuum subspecies Cayenna in olive oil) and Arabic gum in an aqueous solution of extracts of Salvia guaranitica, and Form B, an aqueous solution of extracts of Taxus baccata. Both Form A and B showed to be toxic against P. spumarius compared to the water control. The mean percentage of spittlebug mortality obtained with Form A one hour after treatments was dose-dependent; the lethal dose values were 0.13% (LD25), 0.36% (LD50), and 0.85% (LD75). At the same time, no significant differences in mortality rate were observed between the 0.75% treatment and the treatments with deltamethrin (about 90%). The mean percentage of spittlebug mortality obtained with Form B ranged from 21% to 53% one hour after treatment, but these values were significantly lower than those obtained with deltamethrin. The effectiveness of Form A on the P. spumarius population was also evaluated in the field. The averages of captures in the three experimental blocks were 1.8/trap for treated and 7.7/trap for untreated plots, and the spittlebug populations significantly decreased after treatments. Based on these results and the literature data, we hypothesize that the effectiveness of Form A is the result of the synergistic effect of all its components. No symptoms of phytotoxicity were recorded on olive trees treated with Form A, and the number of P. spumarius specimens collected on these plants was much lower than on untreated plants. These results suggest the potential use of Form A in the protection of olive trees. This vegetal formulation can thus be considered as a valid alternative to chemical insecticides for the control of the main vector of X. fastidiosa and could be integrated into a sustainable management system for OQDS.

Xylella fastidiosa, Possible New Threat to Chestnut (Castanea sativa Mill.) in Italy

This is the first worldwide report of X. fastidiosa (Xf) subsp. pauca on Castanea sativa and the first characterization of Xf infection on this species. Plants located in three sites (in a long-term affected area in Apulia) were monitored for symptoms and bacterial concentrations in spring and summer, while microscopic analyses were performed to evaluate the pathogen distribution in the xylem vessels. All chestnut plants appeared asymptomatic but the Xf subsp. pauca strain “De Donno” was present at low concentrations (1.14 × 104 and 1.56 × 103 cfu mL−1 in April and June) and with a low incidence (respectively, 38% and 30%). The FISH-CLSM (Fluorescent In Situ Hybridization—Confocal Laser Scanning Microscope) analysis showed evident Xf occlusions but in a low percentage (9.2 ± 3.4%); these data can explain the lack of symptoms on the canopy. Furthermore, through a bibliographic analysis it emerged that Philaenus spumarius and Neophilaenus campestris, two Xf vectors present in Europe, are suckling feeding insects on chestnut trees and could be involved in the bacterial transmission to this species. Asymptomatic Xf host species can play a considerable role in new outbreak emergence or in the expansion of existing ones. So, it is essential to identify them to plan more effective monitoring activities.

High Growing Season Temperatures Limit Winter Recovery of Grapevines from Xylella fastidiosa Infection - Implications for Epidemiology in Hot Climates.

Management of widespread plant pathogens is challenging as climatic differences among crop-growing regions may alter key aspects of pathogen spread and disease severity. Xylella fastidiosa is a xylem-limited bacterial pathogen that is transmitted by xylem sap-feeding insects. Geographic distribution of X. fastidiosa is limited by winter climate, and vines infected with X. fastidiosa can recover from infection when held at cold temperatures. California has a long history of research on Pierce's disease and significant geographic and climatic diversity among grape-growing regions. This background in combination with experimental disease studies under controlled temperature conditions can inform risk assessment for X. fastidiosa spread and epidemic severity across different regions and under changing climate conditions. California's grape-growing regions have considerable differences in summer and winter climate. In northern and coastal regions, summers are mild and winters are cool, conditions which favor winter recovery of infected vines. In contrast, in inland and southern areas, summers are hot and winters mild, reducing likelihood of winter recovery. Here, winter recovery of three table grape cultivars (Flame, Scarlet Royal, and Thompson Seedless) and three wine grape cultivars (Sauvignon Blanc, Cabernet Sauvignon, and Zinfandel) were evaluated under temperature conditions representative of the San Joaquin Valley, an area with hot summers and mild winters that has been severely impacted by Pierce's disease and contains a large portion of California grape production. Mechanically inoculated vines were held in the greenhouse under one of three warming treatments to represent different seasonal inoculation dates prior to being moved into a cold chamber. Winter recovery under all treatments was generally limited but with some cultivar variation. Given hot summer temperatures of many grape-growing regions worldwide, as well as increasing global temperatures overall, winter recovery of grapevines should not be considered a key factor limiting X. fastidiosa spread and epidemic severity in the majority of cases.

Draft genome sequence of Xylella fastidiosa strain ATCC 35874 isolated from infected red oak in Washington, DC.

We report here the draft genome sequence of Xylella fastidiosa strain ATCC 35874. The strain was originally isolated from infected red oak in Washington, DC, and obtained from the American Type Culture Collection. The ATCC 35874 genome contains 2,543,332 bp and has a G + C content of 51.72%.

A high‐throughput analysis of high‐resolution X‐ray CT images of stems of olive and citrus plants resistant and susceptible to Xylella fastidiosa

AbstractThe bacterial plant pathogen Xylella fastidiosa causes disease in several globally important crops. However, some cultivars harbour reduced bacterial loads and express few symptoms. Evidence considering plant species in isolation suggests xylem structure influences cultivar susceptibility to X. fastidiosa. We test this theory more broadly by analysing high‐resolution synchrotron X‐ray computed tomography of healthy and infected plant vasculature from two taxonomic groups containing susceptible and resistant varieties: two citrus cultivars (sweet orange cv. Pera, tangor cv. Murcott) and two olive cultivars (Koroneiki, Leccino). Results found the susceptible plants had more vessels than resistant ones, which could promote within‐host pathogen spread. However, features associated with resistance were not shared by citrus and olive. While xylem vessels in resistant citrus stems had comparable diameters to those in susceptible plants, resistant olives had narrower vessels that could limit biofilm spread. And while differences among olive cultivars were not detected, results suggest greater vascular connectivity in resistant compared to susceptible citrus plants. We hypothesize that this provides alternate flow paths for sustaining hydraulic functionality under infection. In summary, this work elucidates different physiological resistance mechanisms between two taxonomic groups, while supporting the existence of an intertaxonomical metric that could speed up the identification of candidate‐resistant plants.

An Unprecedented Experience of Collective Bereavement: The Story of Xylella Fastidiosa in Apulia

An Unprecedented Experience of Collective Bereavement: The Story of Xylella Fastidiosa in Apulia

Permethylation as a strategy for high-molecular-weight polysaccharide structure analysis by nuclear magnetic resonance-Case study of Xylella fastidiosa extracellular polysaccharide.

Current practices for structural analysis of extremely large-molecular-weight polysaccharides via solution-state nuclear magnetic resonance (NMR) spectroscopy incorporate partial depolymerization protocols that enable polysaccharide solubilization in suitable solvents. Non-specific depolymerization techniques utilized for glycosidic bond cleavage, such as chemical degradation or ultrasonication, potentially generate structural fragments that can complicate complete and accurate characterization of polysaccharide structures. Utilization of appropriate enzymes for polysaccharide degradation, on the other hand, requires prior structural knowledge and optimal enzyme activity conditions that are not available to an analyst working with novel or unknown compounds. Herein, we describe an application of a permethylation strategy that allows the complete dissolution of intact polysaccharides for NMR structural characterization. This approach is utilized for NMR analysis of Xylella fastidiosa extracellular polysaccharide (EPS), which is essential for the virulence of the plant pathogen that affects multiple commercial crops and is responsible for multibillion dollar losses each year.

European Grapevine Cultivars and Rootstocks Show Differential Resistance to Xylella fastidiosa Subsp. fastidiosa

Several Xylella fastidiosa subsp. fastidiosa (ST1) strains that cause Pierce’s disease were isolated from grapevine in Spain. In this study, we applied an approach to assess PD susceptibility among 24 different well-known Vitis vinifera subsp. vinifera cultivars and five rootstocks belonging to different species of the genus Vitis. Both were commonly commercialized, representing about 75% of the cultivated area in Spain. This method incorporated disease severity, disease progression, and water potential from the stem xylem. The trials were carried out under field and greenhouse conditions. The virulence of the Xff strain XYL 2055/17 was significantly higher than that of strain XYL 2177/18. However, while this difference in strain virulence did not seem to modify the susceptibility profiles of the cultivars, disease severity could be climate dependent. This work established two significantly different groups of European cultivars of grapevine characterized by high and low susceptibility to Xff ST1: cultivars with high susceptibility, including reference cultivars such as Tempranillo and Tempranillo Blanco, and cultivars with high resistance, such as Hondarrabi Zuri and Cabernet Sauvignon. Cultivar susceptibility was independent of the rootstock on which they were grafted. No conclusive data were found regarding the potential of water loss as an early detection test prior to symptom onset. This study provides a framework with which to advance cultivar susceptibility studies under different environmental conditions.

Transcriptome and Secretome Analyses of Endophyte Methylobacterium mesophilicum and Pathogen Xylella fastidiosa Interacting Show Nutrient Competition.

Xylella fastidiosa is the causal agent of several plant diseases affecting fruit and nut crops. Methylobacterium mesophilicum strain SR1.6/6 was isolated from Citrus sinensis and shown to promote plant growth by producing phytohormones, providing nutrients, inhibiting X. fastidiosa, and preventing Citrus Variegated Chlorosis. However, the molecular mechanisms involved in the interaction among these microbes are still unclear. The present work aimed to analyze physiological and molecular aspects of M. mesophilicum SR1.6/6 and X. fastidiosa 9a5c in co-culture. The transcriptome and secretome analyses indicated that X. fastidiosa down-regulates cell division and transport genes and up-regulates stress via induction of chaperones and pathogenicity-related genes including, the lipase-esterase LesA, a protease, as well as an oligopeptidase in response to M. mesophilicum competition. On the other hand, M. mesophilicum also down-regulated transport genes, except for iron uptake, which was up-regulated. Secretome analysis identified four proteins in M. mesophilicum exclusively produced in co-culture with X. fastidiosa, among these, three are related to phosphorous uptake. These results suggest that M. mesophilicum inhibits X. fastidiosa growth mainly due to nutrient competition for iron and phosphorous, thus promoting X. fastidiosa starvation, besides producing enzymes that degrade X. fastidiosa cell wall, mainly hydrolases. The understanding of these interactions provides a direction for control and management of the phytopathogen X. fastidiosa, and consequently, helps to improve citrus growth and productivity.

Non-Targeted Spectranomics for the Early Detection of Xylella fastidiosa Infection in Asymptomatic Olive Trees, cv. Cellina di Nardò.

Olive quick decline syndrome (OQDS) is a disease that has been seriously affecting olive trees in southern Italy since around 2009. During the disease, caused by Xylella fastidiosa subsp. pauca sequence type ST53 (Xf), the flow of water and nutrients within the trees is significantly compromised. Initially, infected trees may not show any symptoms, making early detection challenging. In this study, young artificially infected plants of the susceptible cultivar Cellina di Nardò were grown in a controlled environment and co-inoculated with additional xylem-inhabiting fungi. Asymptomatic leaves of olive plants at an early stage of infection were collected and analyzed using nuclear magnetic resonance (NMR), hyperspectral reflectance (HSR), and chemometrics. The application of a spectranomic approach contributed to shedding light on the relationship between the presence of specific hydrosoluble metabolites and the optical properties of both asymptomatic Xf-infected and non-infected olive leaves. Significant correlations between wavebands located in the range of 530-560 nm and 1380-1470 nm, and the following metabolites were found to be indicative of Xf infection: malic acid, fructose, sucrose, oleuropein derivatives, and formic acid. This information is the key to the development of HSR-based sensors capable of early detection of Xf infections in olive trees.

Current Status of the Main Olive Pests: Useful Integrated Pest Management Strategies and Genetic Tools

Mediterranean olive cultivation faces challenges in the global environmental change context. Pests and diseases caused by arthropods such as Bactrocera oleae, Prays oleae, and certain vectors of Xylella fastidiosa are expected to increase and spread in part due to this global scenario. The control of these arthropods has relied on synthetic pesticides, the misuse of which has led to pest population resistance and concerns about their negative impacts on biodiversity and global health. Integrated pest management (IPM) methods have emerged through the careful consideration of all available control techniques and the subsequent integration of appropriate measures that discourage the development of pest populations. This paper reviews the IPM guidelines for olive cultivation, prioritizing the use of biological control methods, and the integration of genetics and biotechnology, which bring precision, efficacy, and safety. It evidences the importance of genetic analysis in pest populations, pesticide resistance and in the contributions of predators to pest control. Advances in formulations and delivery systems for pesticides such as Bacillus thuringiensis, plant-incorporated protectants, improved SIT techniques, and the specific efficacy of biologicals pesticides are covered. Finally, this paper explores promising tools such as RNAi and gene drive while recognizing the ethical, environmental, and regulatory challenges associated with their use. Shortly, these innovations have the potential to reduce the environmental impacts of pests while ensuring the long-term viability of the olive industry.

Revealing traits of phytopathogenic status induced by Xylella Fastidiosa in olive trees by analysing multifractal and informational patterns of MODIS satellite evapotranspiration data

Revealing traits of phytopathogenic status induced by Xylella Fastidiosa in olive trees by analysing multifractal and informational patterns of MODIS satellite evapotranspiration data

The Effect of Accuracy and Uncertainty in Test Performance on Optimal Early Detection Survey Budget Allocations

Early detection of pests at a low population prevalence is a key pillar of surveillance for plant health. A novel pest is expected to undergo exponential growth when it first occurs in a host population. This allows for the use of mathematical rules of thumb for the number of samples required to detect presence before it reaches a target prevalence given the diagnostic performance of the tests used. Previous work assumes that pest presence is diagnosed by applying a single diagnostic test to each sample. However, diagnostic decisions are often made based on outcomes of multiple tests applied to the same sample in a consistent test program; for example, an assessment of symptoms followed by a PCR test applied to samples for which symptoms were observed. Each test can have different testing costs, as well as distinct and independent diagnostic performance and uncertainty values. A framework to optimize early detection surveys by minimizing overall costs for test programs that apply up to three diagnostic tests has been developed. The framework assesses the consequences of the test order and logical rule by which diagnoses are determined on costs across a range of pest prevalence. Explicit definitions of uncertainty in key parameters are incorporated to assess the consequences of uncertainty about their true value. Use of the framework is exemplified with two hypothetical case studies exploring the potential impact of selecting a suboptimal test program (based on Xylella fastidiosa) and investment in test improvements (based on lateral flow devices applied to Phytophthora pluvialis). [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Ulía

Ulía is the word for olive in the Salento area of Italy. Ulía is also the word meaning’ I’d like to/ love to’, in the local dialect. The linguistic assonances are only one of the many dimensions that link olive trees and humanity, past and future, nature and desire. Recently, Salento’s hundreds-years-old olive trees have been attacked by a disease called Xylella Fastidiosa. In this paper, I analyze the presence and role of both Pagan and non-Pagan ‘neo-animist’ political discourses oriented toward the cure, preservation, tutelage and guardianship of Salento’s olive trees under threat of felling and eradication.

Xylella fastidiosa subsp. pauca, Neofusicoccum spp. and the Decline of Olive Trees in Salento (Apulia, Italy): Comparison of Symptoms, Possible Interactions, Certainties and Doubts.

Xylella fastidiosa subsp. pauca (XFP), Neofusicoccum mediterraneum, N. stellenboschiana and other fungi have been found in olive groves of Salento (Apulia, Italy) that show symptoms of severe decline. XFP is well known to be the cause of olive quick decline syndrome (OQDS). It has also been assessed that Neofusicoccum spp. causes a distinct disease syndrome, namely, branch and twig dieback (BTD). All these phytopathogens incite severe symptoms that can compromise the viability of large canopy sectors or the whole tree. However, their specific symptoms are not easily distinguished, especially during the final stages of the disease when branches are definitively desiccated. By contrast, they can be differentiated during the initial phases of the infection when some facets of the diseases are typical, especially wood discoloration, incited solely by fungi. Here, we describe the typical symptomatological features of OQDS and BTD that can be observed in the field and that have been confirmed by Koch postulate experiments. Similar symptoms, caused by some abiotic adverse conditions and even by additional biotic factors, are also described. Thus, this review aims at: (i) raising the awareness that declining olive trees in Salento do not have to be linked a priori to XFP; (ii) defining the guidelines for a correct symptomatic diagnosis to orient proper laboratory analyses, which is crucial for the application of effective control measures. The possibility that bacterium and fungi could act as a polyspecies and in conjunction with predisposing abiotic stresses is also widely discussed.

High-quality full genome assembly of historic Xylella fastidiosa strains from ICMP collection using a hybrid sequencing approach.

High-quality complete genomes of five Xylella fastidiosa strains were assembled by combining Nanopore and Illumina sequencing data. Among these, International Collection of Micro-organisms from Plants (ICMP) 8731, ICMP 8742 and ICMP 8745 belong to subspecies fastidiosa while ICMP 8739 and ICMP 8740 were determined as subspecies multiplex. The strains were further classified into sequence types.

Xylella fastidiosa modulates exopolysaccharide polymer length and the dynamics of biofilm development with a β-1,4-endoglucanase.

It is well established that exopolysaccharide (EPS) is an integral structural component of bacterial biofilms necessary for assembly and maintenance of the three-dimensional architecture of the biofilm. However, the process and role of EPS turnover within a developing biofilm is not fully understood. Here, we demonstrated that Xylella fastidiosa uses a self-produced endoglucanase to enzymatically process its own EPS to modulate EPS polymer length. This enzymatic processing of EPS dictates the early stages of X. fastidiosa's biofilm development, which, in turn, affects its behavior in planta. A deletion mutant that cannot produce the endoglucanase was hypervirulent, thereby linking enzymatic processing of EPS to attenuation of virulence in symptomatic hosts, which may be a vestige of X. fastidiosa's commensal behavior in many of its other non-symptomatic hosts.

Evidence of Xylella fastidiosa Infection and Associated Thermal Signatures in Southern Highbush Blueberry (Vaccinium corymbosum Interspecific Hybrids).

Xylella fastidiosa, a gram-negative bacterium vectored to plants via feeding of infected insects, causes a number of notorious plant diseases throughout the world, such as Pierce's disease (grapes), olive quick decline syndrome, and coffee leaf scorch. Detection of Xf in infected plants can be challenging because the early foliar disease symptoms are subtle and may be attributed to multiple minor physiological stresses and/or borderline nutrient deficiencies. Furthermore, Xf may reside within an infected plant for one or more growing seasons before traditional visible diagnostic disease symptoms emerge. Any method that can identify infection during the latent period or pre-diagnostic disease progress state could substantially improve the outcome of disease control interventions. Because Xf locally and gradually impairs water movement through infected plant stems and leaves over time, infected plants may not be able to effectively dissipate heat through transpiration-assisted cooling, and this heat signature may be an important pre-diagnostic disease trait. Here, we report on the association between thermal imaging, the early stages of Xf infection, and disease development in blueberry plants, and discuss the benefits and limitations of using thermal imaging to detect bacterial leaf scorch of blueberries.