Erwinia amylovora is a quarantine phytopathogenic bacterium that is the causal agent of fire blight, a destructive disease responsible for killing millions of fruit-bearing plants worldwide, including apple, pear, quince, and raspberry. Efficient and sustainable control strategies for this serious bacterial disease are still lacking, and traditional methods are limited to the use of antibiotics and some basic agricultural practices. This study aimed to contribute to the development of a sustainable control strategy through the identification, characterization, and application of bacteriophages (phages) able to control fire blight on pears. Phages isolated from wastewater collected in the Apulia region (southern Italy) were characterized and evaluated as antibacterial agents to treat experimental fire blight caused by E. amylovora. Transmission electron microscopy (TEM) conducted on purified phages (named EP-IT22 for Erwinia phage IT22) showed particles with icosahedral heads of ca. 90 ± 5 nm in length and long contractile tails of 100 ± 10 nm, typical of the Myoviridae family. Whole genome sequencing (WGS), assembly, and analysis of the phage DNA generated a single contig of 174.346 bp representing a complete circular genome composed of 310 open reading frames (ORFs). EP-IT22 was found to be 98.48% identical to the Straboviridae Erwinia phage Cronus (EPC) (GenBank Acc. n° NC_055743) at the nucleotide level. EP-IT22 was found to be resistant to high temperatures (up to 60 °C) and pH values between 4 and 11, and was able to accomplish a complete lytic cycle within one hour. Furthermore, the viability-qPCR and turbidity assays showed that EP-IT22 (MOI = 1) lysed 94% of E. amylovora cells in 20 h. The antibacterial activity of EP-IT22 in planta was evaluated in E. amylovora-inoculated pear plants that remained asymptomatic 40 days post inoculation, similarly to those treated with streptomycin sulphate. This is the first description of the morphological, biological, and molecular features of EP-IT22, highlighting its promising potential for biocontrol of E. amylovora against fire blight disease.

Climate and trade changes are reshaping the cartographic distribution of lethal pervasive pathogens. Among serious emerging challenges is Xylella fastidiosa (Xf), a xylem-limited phytopathogenic bacterium that produces losses and damages to numerous crops of high economic and agronomic importance. Lately, this grave quarantine pathogen has expended its distribution by arriving to several European countries and infecting both wild and cultivated plants, and no cure has been identified so far. Countries without current outbreaks like Morocco, need to monitor theirs crops frequently because detecting diseases in the early stages may reduce the huge losses caused by Xf. For that purpose, inspections were managed in different regions in Morocco from March 2020 to July 2021 to assess the presence of Xf in several growing areas of vulnerable economic crops (i.e., almond, citrus and olive). To extend the likelihood of detection, hosts have been inspected and sampled randomly over different environments including symptomatic and asymptomatic plants. Each sample was screened for the existence of Xf by using the DAS-ELISA commercial kit, while, further analyses were carried out for doubtful samples, by PCR. Results of both tests did not show any positive sample in the investigated areas. This finding is an update on the Xf situation in Morocco and confirms that this country is still a free territory from this bacterium, at least in the monitored regions.

Antimicrobial peptides (AMPs) are a various group of molecules found in a wide range of organisms and act as a defense mechanism against different kinds of infectious pathogens (bacteria, viruses, and fungi, etc.). This study explored the antibacterial activity of nine candidates reported in the literature for their effect on human and animal bacteria, (i.e., Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa) against Erwinia amylovora (E. amylovora), the causal agent of fire blight disease on pome fruits. The antibacterial activity of these peptides against E. amylovora was evaluated in vitro using viable-quantitative PCR (v-qPCR), fluorescence microscopy (FM), optical density (OD), and transmission electron microscopy (TEM), while the in vivo control efficacy was evaluated in treating experimental fire blight on pear fruits. With a view to their safe and ecofriendly field use in the future, the study also used animal and plant eukaryotic cells to evaluate the possible toxicity of these AMPs. Results in vitro showed that KL29 was the most potent peptide in inhibiting E. amylovora cell proliferation. In addition, the results of v-qPCR, FM, and TEM showed that KL29 has a bifunctional mechanism of action (lytic and non-lytic) when used at different concentrations against E. amylovora. KL29 reduced fire blight symptoms by 85% when applied experimentally in vivo. Furthermore, it had no impact on animal or plant cells, thus demonstrating its potential for safe use as an antibacterial agent. This study sheds light on a new and potent antibacterial peptide for E. amylovora and its modes of action, which could be exploited to develop sustainable treatments for fire blight.

Xylella fastidiosa (Xf) is a xylem-limited quarantine plant bacterium and one of the most harmful agricultural pathogens across the world. Despite significant research efforts, neither a direct treatment nor an efficient strategy has yet been developed for combatting Xylella-associated diseases. Antimicrobial peptides (AMPs) have been gaining interest as a promising sustainable tool to control pathogens due to their unique mechanism of action, broad spectrum of activity, and low environmental impact. In this study, we disclose the bioactivity of nine AMPs reported in the literature to be efficient against human and plant pathogen bacteria, i.e., Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, against Xf, through in vitro and in vivo experiments. Based on viable-quantitative PCR (v-qPCR), fluorescence microscopy (FM), optical density (OD), and transmission electron microscopy (TEM) assays, peptides Ascaphin-8 (GF19), DASamP1 (FF13), and DASamP2 (IL14) demonstrated the highest bactericidal and antibiofilm activities and were more efficient than the peptide PB178 (KL29), reported as one of the most potent AMPs against Xf at present. Furthermore, these AMPs showed low to no toxicity when tested on eukaryotic cells. In in planta tests, no Xf disease symptoms were noticed in Nicotiana tabacum plants treated with the AMPs 40 days post inoculation. This study highlighted the high antagonistic activity of newly tested AMP candidates against Xf, which could lead to the development of promising eco-friendly management of Xf-related diseases.

Morocco belongs to the countries ranked at a high-risk level for entry, establishment, and spread of Xylella fastidiosa, which has recently re-emerged as a plant pathogen of global importance causing olive quick decline syndrome (OQDS). Symptomatic infection by X. fastidiosa leads to devastating diseases and important economic losses. To prevent such losses and damages, countries without current outbreaks like Morocco need to first understand their host plant responses to X. fastidiosa. The assessment of the macro and micro-elements content (ionome) in leaves can give basic and useful information along with being a powerful tool for the sustainable management of diseases caused by this devastating pathogen. Herein, we compare the leaf ionome of four important autochthonous Moroccan olive cultivars (‘Picholine Marocaine’, ‘Haouzia’, ‘Menara’, and ‘Meslalla’), and eight Mediterranean varieties introduced in Morocco (‘Arbequina’, ‘Arbosana’, ‘Leccino’, ‘Ogliarola salentina’, ‘Cellina di Nardo’, ‘Frantoio’, ‘Leucocarpa’, and ‘Picholine de Languedoc’), to develop hypotheses related to the resistance or susceptibility of the Moroccan olive trees to X. fastidiosa infection. Leaf ionomes, mainly Ca, Cu, Fe, Mg, Mn, Na, Zn, and P, were determined using inductively coupled plasma optical emission spectroscopy (ICP-OES). These varieties were also screened for their total phenolics and flavonoids content. Data were then involved in a comparative scheme to determine the plasticity of the pathogen. Our results showed that the varieties ‘Leccino’, ’Arbosana’, ‘Arbequina’ consistently contained higher Mn, Cu, and Zn and lower Ca and Na levels compared with the higher pathogen-sensitive ‘Ogliarola salentina’ and ‘Cellina di Nardò’. Our findings suggest that ‘Arbozana’, ‘Arbiquina’, ‘Menara’, and ‘Haouzia’ may tolerate the infection by X. fastidiosa to varying degrees, provides additional support for ‘Leccino’ having resistance to X. fastidiosa, and suggests that both ‘Ogliarola salentina’ and ‘Cellina di Nardö’ are likely sensitive to X. fastidiosa infection.