Search Results (11)

Particle film technology is an environmentally sustainable crop protection method, offering an alternative to chemical pesticides for disease control. Copper-based compounds have long been central to the management of bacterial and fungal diseases, particularly in organic agriculture. However, due to their environmental persistence, their use has been increasingly restricted by European regulations, making the management of widespread diseases such as Olive Knot (Pseudomonas savastanoi pv. savastanoi) and Downy Mildew (Plasmopara viticola) more difficult. The LIFE Microfighter project addresses this problem by testing a novel Zeo-Biopesticide (ZBp), in which natural zeolite serves as a carrier for the beneficial bacterium Pseudomonas synxantha DLS65. Field trials conducted in high-rainfall areas of Emilia-Romagna (Italy) evaluated the product’s distribution and persistence on olive and grape leaves through ESEM (Environmental Scanning Electron Microscopy) observations, its ability to retain the microorganism, and its effectiveness for disease control. Results showed that ZBp significantly reduced Olive Knot incidence compared to both the untreated control and Cu-based treatments (p < 0.05), supporting its potential as an alternative for bacterial disease management, while showing no statistically significant difference compared to the control in either the incidence or severity of Downy Mildew (p > 0.05). Its persistence and adherence to plant surfaces, which could influence its overall field performance, were affected by environmental conditions, particularly rainfall. Full article

Olive knot disease, caused by Pseudomonas savastanoi pv. savastanoi, severely impacts olive tree yield and oil quality. Early and accurate detection of the bacterium’s presence, particularly in asymptomatic plants, is crucial for effective disease management. This study aimed to develop an improved protocol for processing plant samples and adapting quantitative PCR to droplet digital PCR (ddPCR). For this purpose, four plant preparations—EW (external washing), PELLET (bacterial concentration), and enrichment in liquid media for 24 or 48 h (24hE, 48hE)—were tested using spiked samples. The ddPCR was set up and compared with qPCR to evaluate analytical sensitivity and specificity. Additionally, field samples from symptomatic and asymptomatic olive orchards were tested to evaluate the performance of the selected methods in naturally infected plants. ddPCR showed higher sensitivity than qPCR, particularly with the PELLET and 24hE preparations. The PELLET from the spiked sample preparation achieved a limit of detection of 10 CFU/mL for both molecular tests. The ddPCR, combined with the PELLET preparation, offers a highly sensitive and reliable tool for detecting P. savastanoi pv. savastanoi in asymptomatic olive material. This protocol shows great potential for improving early bacterial detection and disease prevention, thus aiding control strategies in nurseries and olive orchards, and supporting the production of certified plant propagation material. Full article

One of the most persistent and damaging diseases in olive trees is olive knot disease. This disease is caused by an infection by the Gram-negative phytopathogenic bacterium Pseudomonas savastanoi pv. savastanoi that is notoriously difficult to control. The increasing demand for eco-friendly and sustainable agricultural solutions has driven research into plant-based agents. This study investigated the antibacterial properties of essential oils (EOs) and their constituents, olive mill wastewater (OMWW), the phenolic compound hydroxytyrosol (HTyr), and algae and garlic extracts, as well as copper-based and plant-stimulating commercial products against P. savastanoi pv. savastanoi, a significant olive tree pathogen. Antibacterial activity was determined using the Kirby–Bauer disc diffusion and broth microdilution methods. The EOs derived from Thymus vulgaris (thyme) and Origanum compactum (oregano), and their key components thymol and carvacrol, exhibited the strongest antibacterial efficacy. Conversely, the OMWW, plant-stimulating products, and algae and garlic extracts showed limited to no antibacterial activity in vitro, with their antibacterial properties determined using the disc diffusion method. While the EOs were highly effective in vitro, regardless of the testing method, their efficacy in bacterial growth inhibition was strain- and concentration-dependent, possibly highlighting some metabolic or genetic variability in the target pathogen, even though the MIC values against all tested strains of P. savastanoi pv. savastanoi were equal. Bacterial membrane disruption and the consequent leakage of metabolites were determined as the modes of action of carvacrol and oregano EO. Carvacrol also promoted plant growth in lettuce without significant phytotoxic effects, although minor necrotic lesions were observed in young olive leaves at higher concentrations, presenting these agents as potential next-generation green bactericides. Full article

Super high-density (SHD) planting systems are very efficient in terms of production and water use. In these orchards, water-saving irrigation strategies are used precisely to keep the best sustainability compared to traditional orchards. With agro-climatic and eco-physiological parameter monitoring, the SHD planting system has become even more efficient. SHD orchards, however, could also be more so affected by other pests and diseases than traditional systems, but field responses are still unknown when olive trees are grown in SHD groves. The goal of this two-year field research was to investigate the seasonal changes of the phytosanitary status of ‘Arbosana’ grown in an SHD orchard under both regulated deficit (RDI) and full irrigation regimes (FI). This study investigated the influence of the two different irrigation regimes on the infections of three olive tree key diseases: cercosporiosis (Pseudocercospora cladosporioides), cycloconium (Fusicladium oleagineum), and olive knot (Pseudomonas savastanoi pv. savastanoi). RDI significantly reduced the severity of cercosporiosis in 2020 compared to FI but not in 2021. Cycloconium was observed only as a latent infection during the two studied years and olive knot was not influenced by irrigation but only by weather conditions. These findings suggest that irrigation management can play a key role in controlling cercosporiosis effectively in SHD olive orchards, but also that weather conditions have an even higher impact on the tree key diseases. Full article

Olive knot disease, caused by the bacterium Pseudomonas savastanoi pv. savastanoi, causes great damage in olive orchards. While control measures of P. savastanoi pv. savastanoi in olive orchards primarily rely on pruning and copper-based treatments, the use of antibiotics as bactericidal preparations in agriculture is limited and highly restricted. However, plants are naturally endowed with protective molecules, such as phenolic compounds, which defend them against herbivores, insects, and microorganisms. This research aimed to test the virulence of five strains of P. savastanoi pv. savastanoi isolated from different growing regions and olive varieties, and to examine whether there is a difference in plant susceptibility based on the variety. An additional goal was to test the antimicrobial activity of olive mill wastewater, known for its high content of phenolic compounds, and aqueous garlic hydrolysate, as well as to compare them with a commercial copper-based product, pure hydroxytyrosol, and a standard antibiotic as references. Analysis of knot characteristics showed variations in the virulence of the P. savastanoi pv. savastanoi strains, with the highest virulence being observed for the strain I7L and the lowest virulence for the strain B45C-PR. The olive cultivar Rosinjola displayed higher susceptibility compared to Frantoio, Buža, and Leccino, while cv. Istarska bjelica exhibited the least susceptibility compared to the other investigated olive cultivars. In an attempt to explore alternative solutions for disease control, in vitro tests revealed that the phenol HTyr, GE, and the wastewater with the highest total phenolic content (cv. Istarska bjelica) possess the highest antibacterial activity. This supports the role of polyphenols in host defense, aligning with previous field observations of lower susceptibility of cv. Istarska bjelica to olive knot disease. These findings highlight the complex nature of olive knot interactions with bacterial strains and olive cultivars, simultaneously accentuating and underscoring the importance of considering the host’s defenses as well as bacterial virulence in disease management strategies. Full article

Essential oil-based pesticides, which contain antimicrobial and antioxidant molecules, have potential for use in sustainable agriculture. However, these compounds have limitations such as volatility, poor water solubility, and phytotoxicity. Nanoencapsulation, through processes like micro- and nanoemulsions, can enhance the stability and bioactivity of essential oils. In this study, thyme essential oil from supercritical carbon dioxide extraction was selected as a sustainable antimicrobial tool and nanoencapsulated in an oil-in-water emulsion system. The investigated protocol provided high-speed homogenisation in the presence of cellulose nanocrystals as stabilisers and calcium chloride as an ionic crosslinking agent. Thyme essential oil was characterised via GC-MS and UV-vis analysis, indicating rich content in phenols. The cellulose nanocrystal/essential oil ratio and calcium chloride concentration were varied to tune the nanoemulsions’ physical–chemical stability, which was investigated via UV-vis, direct observation, dynamic light scattering, and Turbiscan analysis. Transmission electron microscopy confirmed the nanosized droplet formation. The nanoemulsion resulting from the addition of crosslinked nanocrystals was very stable over time at room temperature. It was evaluated for the first time on Pseudomonas savastanoi pv. savastanoi, the causal agent of olive knot disease. In vitro tests showed a synergistic effect of the formulation components, and in vivo tests on olive seedlings demonstrated reduced bacterial colonies without any phytotoxic effect. These findings suggest that crosslinked cellulose nanocrystal emulsions can enhance the stability and bioactivity of thyme essential oil, providing a new tool for crop protection. Full article

Plant pathogenic bacteria pose a significant threat to olive cultivation, leading to substantial economic losses and reduced yield. The efficacy of antimicrobial agents against these pathogens is of great interest for sustainable disease management strategies. As such, the management of olive knot disease is one of the major challenges in olive protection. In the presented study, through a series of in vitro assays, we investigated the antimicrobial effect of six essential oils (EOs) and their most concentrated constituents against causative agent of olive knot disease—Pseudomonas savastanoi pv. savastanoi, highlighting the high potential of Origanum compactum EO and its constituent carvacrol. Carvacrol exhibited the highest potential for practical application, demonstrating membrane disruption as its mechanism of action even at the lowest concentration. The bactericidal effect of antimicrobials was confirmed in a time–kill assay, where concentrations of MIC, 2× MIC, and 4× MIC were evaluated. Some of the applied treatments resulted in inhibition equal or higher than copper-based treatment. Additionally, we assessed the phytotoxicity of carvacrol by foliar application on olive cv. Leccino. The appearance of phytotoxic injuries majorly occurred on the young leaves of olive plants, with the highest proportion of damaged canopy observed when the 2× MIC concentration was applied. Due to its great efficiency against P. savastanoi pv. savastanoi in vitro, these findings highlight the potential of carvacrol as a molecule of interest for the development of environmentally friendly biopesticides. This study also contributes to the advancement of disease management practices in olive cultivation, leading to enhanced crop protection. Full article

Olive knot disease (OKD) induced by the bacterium Pseudomonas savastanoi pv. savastanoi seriously affects olive production in the Mediterranean basin. Nowadays, the only strategies to control the disease are pruning and the application of cupric products. An essential strategy to enhance protection is represented by the identification of resistant cultivars, which represents a crucial opportunity for future investments and breeding. We undertook a three-year-long survey at the International Olive Germplasm Collection of “Villa Zagaria” (Sicily, Italy) on thirty-six Sicilian cultivars that were monitored for symptom development. Cultivars with different levels of susceptibility were divided into five clusters. Moreover, in order to investigate possible interactions with endophytic microbial communities, two cultivars with contrasting susceptibilities, Zaituna (highly resistant) and Giarraffa (highly susceptible), were selected for an amplicon-based metagenomic analysis. Distinct endophytic communities colonized the two cultivars, suggesting an interaction between the resident bacterial community and the pathogen. Significantly higher bacterial richness was detected in the shoots of the susceptible cv. Giarraffa, although it had lower diversity. The opposite trend was observed for fungal communities. Among the microbes resulted to be enriched in cv. Giarraffa, it is important to underline the presence of Pseudomonas among the bacterial genera, and Alternaria, Neofusicoccum, Epicoccum, Ascochyta, and Elsinoe among the fungal genera, which include many species often described as plant pathogens and biocontrol agents. Starting from this basic information, new strategies of control, which include breeding for resistance and integrated disease management, can be envisaged. Full article

Strains of Pseudomonas savastanoi pv. savastanoi (Pss), isolated from infected olive trees (Olea europaea L.) in three European countries (Croatia, Slovenia and Portugal) were identified and characterised according to their colony morphology, physiological and biochemical features. According to the LOPAT scheme, 38.6% of Pss isolates were grouped in the Ib cluster. The Portuguese Pss strains were fully consistent with the typical LOPAT profile for this bacterium. Conversely, most Slovenian Pss strains showed delayed oxidase activity, whilst Croatian Pss strains did not produce any fluorescent pigment when grown in vitro. For Pss molecular identification, both end-point and real-time PCR were used, as well as MALDI–TOF, which was additionally used for proteomic analysis and the subsequent species identification of a number of strains that showed deviations from expected LOPAT results. Pss was confirmed as a causal agent of olive knot disease in 46.6% of olive orchards screened. Overall, these data suggests a possible correlation of certain Pss features with the geographical origin and the ecological niche of Pss isolates. Full article

Two bacteria belonging to the Pseudomonas and Pantoea genera were isolated from olive knots. Both bacterial strains were omnipresent in this study’s olive orchard with high susceptibility of the autochthonous olive genotypes indicating coevolution of bacteria with host plants. Genomes of two endemic bacteria show conserved core genomes and genome plasticity. The Pseudomonas ST1 genome has conserved virulence-related genes including genes for quorum sensing, pilus, and flagella biosynthesis, two copies of indole acetic acid biosynthesis (IAA) operons, type I-VI secretions systems, and genes for alginate and levan biosynthesis. Development of knots depends only on the presence of the Pseudomonas ST1 strain which then allows Pantoea paga strain co-infection and cohabitation in developed knots. The two bacteria are sensitive to a large number of antimicrobials, antibiotics, H₂O₂, and Cu (II) salts that can be efficiently used in propagation of bacterial free olive cultivars. Full article

Pseudomonas savastanoi pv. savastanoi is the causal agent of Olive knot disease, relying on the Type Three Secretion System (TTSS) for its pathogenicity. In this regard, nothing was known about the two other pathovars belonging to this species, pv. nerii and pv. fraxini, characterized by a different host range. Here we report on the organization of the entire TTSS cluster on the three pathovars, and a phylogenetic analysis including the TTSS of those bacteria belonging to the P. syringae complex sequenced so far, highlighting the evolution of each operon (hrpC, hrpJ, hrpRS, hrpU and hrpZ). Moreover, by Real-Time PCR we analyzed the in vitro expression of four main TTSS genes, revealing different activation patterns in the three pathovars, hypothetically related to their diverse virulence behaviors. Full article