Draft Whole-Genome Sequences of Xylella fastidiosa subsp. fastidiosa Strains TPD3 and TPD4, Isolated from Grapevines in Hou-li, Taiwan.

We report the draft assemblies of TPD3 and TPD4, two Xylella fastidiosa subsp. fastidiosa isolates infecting grapevines in Hou-li, Taiwan. TPD3 and TPD4 showed similar characteristics regarding genome size (2,483,503 bp and 2,491,539 bp, respectively), GC content (51.49% and 51.47%, respectively), and number of protein-coding sequences (2,394 and 2,413, respectively)

Discriminating between viable and membrane-damaged cells of the plant pathogen Xylella fastidiosa.

Xylella fastidiosa is a plant pathogenic bacterium with devastating consequences to several crops of economic importance across the world. While this pathogen has been studied for over a century in the United States, several aspects of its biology remain to be investigated. Determining the physiological state of bacteria is essential to understand the effects of its interactions with different biotic and abiotic factors on cell viability. Although X. fastidiosa is culturable, its slow growing nature makes this technique cumbersome to assess the physiological state of cells present in a given environment. PMA-qPCR, i.e. the use of quantitative PCR combined with the pre-treatment of cells with the dye propidium monoazide, has been successfully used in a number of studies on human pathogens to calculate the proportion of viable cells, but has less frequently been tested on plant pathogens. We found that the use of a version of PMA, PMAxx, facilitated distinguishing between viable and non-viable cells based on cell membrane integrity in vitro and in planta. Additional experiments comparing the number of culturable, viable, and total cells in planta would help further confirm our initial results. Enhancers, intended to improve the efficacy of PMAxx, were not effective and appeared to be slightly toxic to X. fastidiosa

XatA, an AT-1 autotransporter important for the virulence of Xylella fastidiosa Temecula1.

Xylella fastidiosa Temecula1 is the causative agent of Pierce's disease of grapevine, which is spread by xylem-feeding insects. An important feature of the infection cycle is the ability of X. fastidiosa to colonize and interact with two distinct environments, the xylem of susceptible plants and the insect foregut. Here, we describe our characterization of XatA, the X. fastidiosa autotransporter protein encoded by PD0528. XatA, which is classified as an AT-1 (classical) autotransporter, has a C-terminal β-barrel domain and a passenger domain composed of six tandem repeats of approximately 50 amino acids. Localization studies indicate that XatA is present in both the outer membrane and membrane vesicles and its passenger domain can be found in the supernatant. Moreover, XatA is important for X. fastidiosa autoaggregation and biofilm formation based on mutational analysis and the discovery that Escherichia coli expressing XatA acquire these traits. The xatA mutant also shows a significant decrease in Pierce's disease symptoms when inoculated into grapevines. Finally, X. fastidiosa homologs to XatA, which can be divided into three distinct groups based on synteny, form a single, well-supported clade, suggesting that they arose from a common ancestor

Lipid profile of Xylella fastidiosa Subsp. pauca associated with the olive quick decline syndrome

Lipids, components of the plasma and intracellular membranes as well as of droplets, provide different biological functions related to energy, carbon storage, and stress responses. Bacterial species display diverse membrane composition that changes in response to the different environmental conditions. During plant-pathogen interactions, lipids might have roles in several aspects such as recognition, signal transduction, and downstream responses. Among lipid entities, free fatty acids (FFAs) and their oxidized form, the oxylipins, represent an important class of signaling molecules in host-pathogen perception, especially related to virulence and defense. In bacteria, FFAs (e.g., diffusible signaling factors) and oxylipins have a crucial role in modulating motility, biofilm formation, and virulence. In this study, we explore by LC-TOF and LC-MS/MS the lipid composition of Xylella fastidiosa subsp. pauca strain De Donno in pure culture; some specific lipids (e.g., ornithine lipids and the oxylipin 7,10-diHOME), characteristic of other pathogenic bacteria, were revealed. Nicotiana tabacum was used for testing the ability of this pathogen in producing such lipids in the host. Different lipid compounds present a clear distribution pattern within the infected plant tissues compared to the uninfected ones

Proconiini sharpshooters of Argentina, with notes on its distribution, host plants, and natural enemies

The American tribe Proconiini (Hemiptera: Cicadellidae: Cicadellinae) is one of the largest groups of xylem-feeding insects and includes the majority of the known vectors of xylem-born phytopathogenic organisms. The significance of the pathogens that this group transmits gives them an important role as pests, mostly for citrus fruit, grapes, and almonds. Knowledge of these Hemiptera in Argentina is insufficient and fragmentary. Thus one of the aims of this paper is to summarize the available information of the Proconiini sharpshooters in Argentina. In addition, 14 species are mentioned for the first time in the country, and new distributional data are given for 18 species. Thirty-four new associations between sharpshooters and host plants are recorded. New records of egg parasitoids are given for Dechacona missionum, Molomea consolida, M. lineiceps, and Tapajosa similis.Fil: Paradell, Susana Liria. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. División Entomología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Virla, Eduardo Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Planta Piloto de Procesos Industriales Microbiologicos; ArgentinaFil: Logarzo, Guillermo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. United States Department of Agriculture; Estados UnidosFil: Dellapé, Gimena. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. División Entomología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

The potential direct economic impact and private management costs of an invasive alien species:Xylella fastidiosa on Lebanese wine grapes

Since its outbreak in 2013 in Italy, the harmful bacterium Xylella fastidiosa has continued to spread through-out the Euro-Mediterranean basin and, more recently, in the Middle East region. Xylella fastidiosa subsp. fastidiosa is the causal agent of Pierce’s disease on grapevines. At present, this alien subspecies has not been reported in Lebanon but if this biological invader was to spread with no cost-effective and sustainable management, it would put Lebanese vineyards at a certain level of risk. In the absence of an Xylella fastidiosa subsp. fastidiosa outbreak, the gross revenue generated by Lebanese wine growers is estimated as close to US$22 million/year for an average period of 5 years (2015–2019). The potential quantitative economic impacts of an Xylella fastidiosa subsp. fastidiosa outbreak and particularly, the private control costs have not been assessed yet for this country as well as for others which Xylella fastidiosa may invade. Here, we have aimed to estimate the potential direct economic impact on growers’ livelihoods and provide the first estimate of the private management costs that a theoretical Xylella fastidiosa subsp. fastidiosa outbreak in Lebanon would involve. For this purpose, we used a Partial Budget approach at the farm gate. For the country as a whole, we estimated that a hypothetical full spread of Xylella fastidiosa subsp. fastidiosa on Lebanese wine grapes would lead to maximum potential gross revenue losses of almost US$ 11 million for an average recovery period of4 years, to around US$82.44 million for an average grapevine life span period of 30 years in which infected plants are not replaced at all. The first yearly estimated additional management cost is US$853 per potentially infected hectare. For a recovery period of 4 years, the aggregate estimated additional cost would reach US$2374/ha, while the aggregate net change in profit would be US$-4046/ha. Furthermore, additional work will be needed to estimate the public costs of an Xylella fastidiosa subsp. fastidiosa outbreak in Lebanon. The observed costs in this study support the concerned policy makers and stakeholders to implement a set of reduction management options against Xylella fastidiosa subsp. fastidiosa at both national and wine growers’ levels. This re-emerging alien biota should not be neglected in this country. This understanding of thepotential direct economic impact of Xylella fastidiosa subsp. fastidiosa and the private management costs can also benefit further larger-scale studies covering other potential infection areas and plant hosts

Site Directed Gene Disruption of rpfA in Xylella fastidiosa : Non-Biofilm Producing Mutants as an Approach to Eradication of Pierce\u27s Disease in Grapevine

Xylella fastidious a, a gram-negative, rod shaped bacterium, causes disease in many economically important plants including citrus, coffee, and grapevine. Xylella fastidiosa is xylem limited and is dependent on insect vectors for transmission. Pierce’s Disease of Vitis vinifere grape, transmitted by an insect vector, poses a serious threat to California vineyards today. The glassy-winged sharp-shooter, Homalodisca coagulata, has caused millions of dollars of damage to the California wine industry in just a few years. There is evidence that Xylella fastidiosa may share the molecular mechanisms through which the bacterium establishes infection with a closely related species, Xanthomonas campestris. A cluster of genes designated as regulation of pathogenicity factors are responsible for extracellular polysaccharide production and have been found in the genome of Xanthanomonas campestris and Xylella fastidiosa. Site-directed disruption of regulation of pathogenicity factor A, using homologous recombination in the Xylella fastidiosa strains Temecula and Napa, exhibited a decrease in extracellular polysaccharide that resulted in the inhibition of biofilm formation

An MLSA-based online scheme for the rapid identification of Stenotrophomonas isolates

An online scheme to assign Stenotrophomonas isolates to genomic groups was developed using the multilocus sequence analysis (MLSA), which is based on the DNA sequencing of selected fragments of the housekeeping genes ATP synthase alpha subunit (atpA), the recombination repair protein (recA), the RNA polymerase alpha subunit (rpoA) and the excision repair beta subunit (uvrB). This MLSA-based scheme was validated using eight of the 10 Stenotrophomonas species that have been previously described. The environmental and nosocomial Stenotrophomonas strains were characterised using MLSA, 16S rRNA sequencing and DNA-DNA hybridisation (DDH) analyses. Strains of the same species were found to have greater than 95% concatenated sequence similarity and specific strains formed cohesive readily recognisable phylogenetic groups. Therefore, MLSA appeared to be an effective alternative methodology to amplified fragment length polymorphism fingerprint and DDH techniques. Strains of Stenotrophomonas can be readily assigned through the open database resource that was developed in the current study (www.steno.lncc.br/)

Vitis sp. response to Xylella fastidiosa strain CoDiRO

Following a request from the European Commission, the EFSA Panel on Plant Health assessed a scientific report submitted by the Italian Authorities to the European Commission to support a request to delist Vitissp. from Annex I (‘specified plants’) of the Commission Implementing Decision (EU) 2015/789 of 18 May 2015 to prevent the introduction into and the spread within the Union of Xylella fastidiosa (Wells et al.). The report comprised (i) surveys to detect X. fastidiosa in vineyards located in the epidemic zone of CoDiRO with high numbers of diseased olive trees; (ii) inoculation experiments to infect grapevine with a X. fastidiosa isolate ‘De Donno’ from CoDiRO diseased olives; and (iii) vector transmission experiments with X. fastidiosa infective Philaenus spumarius. The Panel acknowledges the difficulties in providing evidence about this hitherto unknown pathogen/vector/host interaction to support the hypothesis that a plant species cannot be infected with a pathogen. Although field surveys to detect X. fastidiosa in grapevine were negative, there was no supporting information on infective vector populations present in the vineyards. Hence absence of infection pressure cannot be excluded. Furthermore the failure to infect grapevine plants either by artificial inoculation or by vector transmission might be due to inoculation conditions not appropriate to induce infections in grapevine. The detection of X. fastidiosa DNA in inoculated grapevine plants even 12 months after inoculation, although localised at the inoculation points, cannot exclude that the DNA amplified by qPCR was from viable cells. The results presented are coherent and provide converging lines of evidence that grapevine (Vitis vinifera) is not a major susceptible host of X. fastidiosa strain CoDiRO. However, from the experimental evidence it is premature to exclude that systemic infections of V.vinifera and Vitissp. occur and that infections at limited foci could serve as a source of inoculum