Xylella fastidiosa and Drought Stress in Olive Trees: A Complex Relationship Mediated by Soluble Sugars

Xylella fastidiosa (Xf) subsp. pauca “De Donno” is the etiological agent of “Olive Quick Decline Syndrome” (OQDS) on olive trees (Olea europaea L.); the presence of the bacterium causes xylem vessel occlusions inducing a drought stress and the development of leaf scorch symptoms, which may be worsened by water shortage in summer. In order to evaluate how the two stress factors overlap each other, the carbohydrate content and the expression patterns of genes related to carbohydrate metabolism have been evaluated in two olive cvs trees (Cellina di Nardò, susceptible to Xf, and Leccino, resistant to Xf) reporting transcriptional dynamics elicited by Xf infection, drought, or combined stress (drought/Xf). In the Xf-susceptible Cellina di Nardò plants, Xf and its combination with drought significantly decrease total sugars compared to control (−27.0% and −25.7%, respectively). In contrast, the Xf-resistant Leccino plants show a more limited reduction in sugar content in Xf-positive conditions (−20.1%) and combined stresses (−11.1%). Furthermore, while the amount of glucose decreases significantly in stressed Cellina di Nardò plants (≈18%), an increase was observed in Leccino plants under drought/Xf combined stresses (+11.2%). An opposite behavior among cvs was also observed for sucrose, as an accumulation of the disaccharide was recorded in stressed Leccino plants (≈37%). The different response to combined stress by Xf-resistant plants was confirmed considering genes coding for the sucrose or monosaccharide transporter (OeSUT1, OeMST2), the cell wall or vacuolar invertase (OeINV-CW, OeINV-V), the granule-bound starch synthase I (OeGBSSI) and sucrose synthase (OeSUSY), with a higher expression than at least one single stress (e.g., ≈1-fold higher or more than Xf for OeMST2, OeINV-CW, OeINV-V, OeGBSSI). It is probable that the pathways involved in drought stress response induce positive effects useful for pathogen resistance in cv Leccino, confirming the importance of investigating the mechanisms of cross-talk of biotic and abiotic responses.

Doenças associadas à Xylella fastidiosa no Brasil

Based on genomic information, the Xylella fastidiosa bacterium is classified into three main subspecies: fastidiosa, multiplex and pauca. These different subspecies are naturally transmitted through vector insects, known as sharpshooters, which feed on the sap of xylem vessels of plants, where the bacteria colonize. Mainly due to the obstruction in the transport of water and mineral salts in these conducting vessels, the X. fastidiosa bacterium causes serious diseases in different cultures of economic interest. In Brazil, this phytopathogen was detected for the first time in plum trees, a crop in which the multiplex subspecies is responsible for the disease known as plum leaf scald. Subsequently, the pauca subspecie was associated with citrus variegated chlorosis in sweet orange orchards in São Paulo State. In the same state, the pauca subspecie was also found in coffee plants causing atrophy of the coffee tree branches, a disease whose damage has not been quantified yet. Recently, the pauca subspecie has also been found in olive trees, causing the olive quick decline syndrome, reported in the southeast region, especially in the Serra da Mantiqueira region. In this review, specific aspects of these diseases were focused, as well as measures that can be adopted as management.

Community Analysis of Culturable Sapwood Endophytes from Apulian Olive Varieties with Different Susceptibility to Xylella fastidiosa

Endophytes are symptomless fungal and/or bacterial microorganisms found in almost all living plant species. The symbiotic association with their host plants by colonizing the internal tissues has endowed them as a valuable tool to suppress diseases, stimulate growth, and promote stress resistance. In this context, the study of culturable endophytes residing the sapwood of Apulian olives might be a promising control strategy for xylem colonizing pathogens as Xylella fastidiosa. To date, olive sapwood cultivable endophytes are still under exploration; therefore, this work pursues a study of cultivable endophytes occurrence variation in the sapwood of different olive varieties under the effect of seasonality, geographical coordinates, and X. fastidiosa infection status. Our study confirms the stability of sapwood endophytic culturable communities in the resistant olive variety and presents the seasonal and geographical fluctuation of olive trees’ sapwood endophytes. It also describes the diversity and occurrence frequency of fungal and bacterial genera, and finally retrieves some of the sapwood-inhabiting fungal and bacterial isolates, known as biocontrol agents of plant pathogens. Thus, the potential role of these bacterial and fungal isolates in conferring olive tree protection against X. fastidiosa should be further investigated.

Physiochemically distinct SU-8 surfaces tailor Xylella fastidiosa cell-surface holdfast and colonization

SU-8 polymer is an excellent platform for diverse applications due to its high aspect ratio of micro/nanostructures fabrication and exceptional optical, chemical, and biocompatible properties. Although SU-8 has been often investigated for a variety of biological applications, how its surface properties influence both the interaction of bacterial cells with the substrate and its colonization is poorly understood. In this work, we tailor SU-8 nanoscale surface properties to investigate single cell motility, adhesion and successive colonization of a phytopathogenic bacteria, Xylella fastidiosa. Different surface properties of SU-8 thin films have been prepared using photolithography processing and oxygen plasma treatment. We found a significant difference in bacterial cell behavior and subsequent colonization on SU-8 as surface property changes. A larger density of carboxyl groups in hydrophilic plasma-treated SU-8 surfaces promotes faster cell motility in the earlier stage of the growth. The hydrophobic nature of pristine SU-8 surfaces has no trackable bacterial motility with 5 to 10 times more single cells adhered to surface than its plasma-treated counterpart. In fact, plasma-treated SU-8 samples suppressed bacterial adhesion, with surfaces showing less than 5% coverage. These results not only showcase that SU-8 surface properties can impact the bacterial behavior in a spatiotemporal manner, but also provide insights on the prominent ability of pathogens to evolve and adapt to different surface properties.

Genetic differentiation of Xylella fastidiosa following the introduction into Taiwan

The economically important plant pathogen Xylella fastidiosa has been reported in multiple regions of the globe during the last two decades, threatening a growing list of plants. Particularly, X. fastidiosa subspecies fastidiosa causes Pierce’s disease (PD) of grapevines, which is a problem in the USA, Spain, and Taiwan. In this work, we studied PD-causing subsp. fastidiosa populations and compared the genome sequences of 33 isolates found in Central Taiwan with 171 isolates from the USA and two from Spain. Phylogenetic relationships, haplotype networks, and genetic diversity analyses confirmed that subsp. fastidiosa was recently introduced into Taiwan from the Southeast USA (i.e. the PD-I lineage). Recent core-genome recombination events were detected among introduced subsp. fastidiosa isolates in Taiwan and contributed to the development of genetic diversity. The genetic diversity observed includes contributions through recombination from unknown donors, suggesting that higher genetic diversity exists in the region. Nevertheless, no recombination event was detected between X. fastidiosa subsp. fastidiosa and the endemic sister species Xylella taiwanensis , which is the causative agent of pear leaf scorch disease. In summary, this study improved our understanding of the genetic diversity of an important plant pathogenic bacterium after its invasion to a new region.

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 throughout 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 of 4 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 the potential 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.

Controlled spatial organization of bacterial growth reveals key role of cell filamentation preceding Xylella fastidiosa biofilm formation

The morphological plasticity of bacteria to form filamentous cells commonly represents an adaptive strategy induced by stresses. In contrast, for diverse human and plant pathogens, filamentous cells have been recently observed during biofilm formation, but their functions and triggering mechanisms remain unclear. To experimentally identify the underlying function and hypothesized cell communication triggers of such cell morphogenesis, spatially controlled cell patterning is pivotal. Here, we demonstrate highly selective cell adhesion of the biofilm-forming phytopathogen Xylella fastidiosa to gold-patterned SiO2 substrates with well-defined geometries and dimensions. The consequent control of both cell density and distances between cell clusters demonstrated that filamentous cell formation depends on cell cluster density, and their ability to interconnect neighboring cell clusters is distance-dependent. This process allows the creation of large interconnected cell clusters that form the structural framework for macroscale biofilms. The addition of diffusible signaling molecules from supernatant extracts provides evidence that cell filamentation is induced by quorum sensing. These findings and our innovative platform could facilitate therapeutic developments targeting biofilm formation mechanisms of X. fastidiosa and other pathogens.

Csp1, a Cold Shock Protein Homolog in Xylella fastidiosa Influences Cell Attachment, Pili Formation, and Gene Expression

Xylella fastidiosa is a major threat to the worldwide agriculture industry (1, 2). Despite its global importance, many aspects of X. fastidiosa biology and pathogenicity are poorly understood.

Antimicrobial Peptides With Antibiofilm Activity Against Xylella fastidiosa

Xylella fastidiosa is a plant pathogen that was recently introduced in Europe and is causing havoc to its agriculture. This Gram-negative bacterium invades the host xylem, multiplies, and forms biofilm occluding the vessels and killing its host. In spite of the great research effort, there is no method that effectively prevents or cures hosts from infections. The main control strategies up to now are eradication, vector control, and pathogen-free plant material. Antimicrobial peptides have arisen as promising candidates to combat this bacterium due to their broad spectrum of activity and low environmental impact. In this work, peptides previously reported in the literature and newly designed analogs were studied for its bactericidal and antibiofilm activity against X. fastidiosa. Also, their hemolytic activity and effect on tobacco leaves when infiltrated were determined. To assess the activity of peptides, the strain IVIA 5387.2 with moderate growth, able to produce biofilm and susceptible to antimicrobial peptides, was selected among six representative strains found in the Mediterranean area (DD1, CFBP 8173, Temecula, IVIA 5387.2, IVIA 5770, and IVIA 5901.2). Two interesting groups of peptides were identified with bactericidal and/or antibiofilm activity and low-moderate toxicity. The peptides 1036 and RIJK2 with dual (bactericidal–antibiofilm) activity against the pathogen and moderate toxicity stand out as the best candidates to control X. fastidiosa diseases. Nevertheless, peptides with only antibiofilm activity and low toxicity are also promising agents as they could prevent the occlusion of xylem vessels caused by the pathogen. The present work contributes to provide novel compounds with antimicrobial and antibiofilm activity that could lead to the development of new treatments against diseases caused by X. fastidiosa.