Search Results (29)

Citrus fruits are major economic and nutritional crops that are sometimes subjected to serious attacks by many fungal phytopathogens after harvesting. In this study, we focus on the structures of potential antifungal nanocomposites from artichoke leaf extract (Art), Art-mediated nanosilver (AgNPs), and their nanoconjugates with chitosan nanoparticles (Cht) to eradicate the blue mold fungus (Penicillium italicum) and preserve oranges during storage via nanocomposite-based edible coatings (ECs). The biosynthesis and conjugation of nanomaterials were verified using UV and infrared (FTIR) spectroscopy, electron microscopy (TEM and SEM) analysis, and DLS assessments. Art could effectually biosynthesize/cap AgNPs with a mean size of 10.35 nm, whereas the average size of Cht was 148.67 nm, and the particles of their nanocomposites had average diameters of 203.22 nm. All nanomaterials/composites exhibited potent antifungal action toward P. italicum isolates; the Cht/Art/AgNP nanocomposite was the most effectual, with an inhibition zone of 31.1 mm and a fungicidal concentration of 17.5 mg/mL, significantly exceeding the activity of other compounds and the fungicide Enilconazole (24.8 mm and 25.0 mg/mL, respectively). The microscopic imaging of P. italicum mycelia treated with Cht/Art/AgNP nanocomposites emphasized their action for the complete destruction of mycelia within 24 h. The orange (Citrus sinensis) fruit coatings, with nanomaterial-based ECs, were highly effectual for preventing blue mold development and preserved fruits for >14 days without any infestation signs; when the control infected fruits were fully covered with blue mold, the infestation remarks covered 12.4%, 5.2%, and 0% of the orange coated with Cht Art/AgNPs and Cht/Art/AgNPs. The constructed Cht/Art/AgNP nanocomposites have potential as effectual biomaterials for protecting citrus fruits from fungal deterioration and preserving their quality. Full article

Blue mold caused by Penicillium italicum (P. italicum) is a major postharvest disease in citrus fruits. Lipoic acid (LA) is a potent antioxidant with biological activity that was evaluated for its inhibitory effects on P. italicum and citrus blue mold using in vitro and in vivo experiments. The results demonstrated that LA effectively suppressed the mycelial growth and spore germination of P. italicum. LA increased hydrogen peroxide levels, compromising cell membrane integrity and leading to enhanced membrane permeability, as indicated by the increased relative conductivity and decreased protein and total sugar contents in P. italicum mycelia. Furthermore, LA delayed disease progression in citrus fruits infected with P. italicum through increasing total phenol and flavonoid contents and enhancing the activities of phenylalanine ammonia lyase, polyphenol oxidase, superoxide dismutase, and peroxidase in citrus peel. Overall, LA exhibited strong antifungal activity against P. italicum and improved citrus fruit resistance to blue mold, highlighting its potential as a natural postharvest disease control agent. Full article

Green mold caused by Penicillium digitatum is a major post-harvest disease in citrus fruits. Therefore, the search for sustainable and low-environmental-impact alternatives for the management of these fungi is of utmost importance. Physalis peruviana L. is a native fruit of the Peruvian Andes with rich bioactive components present throughout the plant. Its antifungal activity stands out, attributed to its high content of phenols, coupled with its antioxidant capacity and antimicrobial activity. Plants were cultivated aeroponically under a combination of red, mixed (50% red, 50% blue), and green LED lights. Additionally, in vitro-habituated roots free of plant growth regulators were also cultivated. An ethanol extraction assisted by ultrasound for 30 min followed by maceration for 72 h was performed, and the extract was filtrated and evaporated in an extraction hood. Antioxidant activity was assessed using the DPPH method, total polyphenols were measured using the Folin–Ciocâlteu method, and an antifungal test in vitro by the poisoned food method was conducted against P. digitatum. In vitro assays revealed that extracts from leaves, roots, and fruits exerted a significant inhibitory effect on the growth of P. digitatum, as evidenced by a reduction in colony radius when cultured employing the poisoned food method, with IC₅₀ values of 62.17, 53.15, and 286.34 µg·mL⁻¹, respectively, compared to 2297 µg·mL⁻¹ for the commercial fungicide Captan 50WP. Although leaves had higher total polyphenol content, no direct correlation with antifungal activity was found. Colored LEDs enhanced phenol accumulation, antioxidant capacity, and antifungal properties in plant parts compared to white LEDs and in vitro roots. These findings suggest P. peruviana as a new alternative biological production system to provide natural compounds for post-harvest disease management. Full article

Blue mold disease, caused by Penicillium italicum (P. italicum), presents a significant challenge to orange fruits (Citrus sinensis L.) and other citrus crops globally. Biological control, particularly Trichoderma species, offers a promising alternative to synthetic fungicides. Therefore, this study aimed to isolate, identify, and evaluate the antagonistic activities of two Trichoderma isolates against P. italicum. These isolates were molecularly identified and assigned accession numbers PP002254 and PP002272, respectively. Both isolates demonstrated significant antifungal activity in dual culture assays. Moreover, the culture filtrates (CFs) of Trichoderma longibrachiatum PP002254 and Trichoderma harzianum PP002272 suppressed the mycelial growth of P. italicum by 77.22% and 71.66%, respectively. Additionally, CFs reduced the severity of blue mold on orange fruits by 26.85% and 53.81%, compared to 100% in the control group. Scanning electron microscopy revealed that treated P. italicum hyphae were shrunken and disfigured. Enzyme activities (catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase) in treated oranges increased, along with total soluble phenolics and flavonoids. Conversely, malondialdehyde (MDA) levels decreased in treated fruits. These findings suggest that T. longibrachiatum PP002254 and T. harzianum PP002272 could be effective biocontrol agents for managing blue mold and other citrus postharvest diseases. Full article

Citrus fruits, particularly mandarins, are highly valued globally for their nutritional benefits and versatile culinary uses. However, the challenge of post-harvest decay, primarily due to blue mold (Penicillium italicum) infections, results in significant food losses and necessitates effective preservation strategies. Traditional methods often rely on fungicides, raising concerns about chemical residues and environmental impact. This study investigates the efficacy of ozone as an alternative approach to controlling blue mold in mandarins. Various gaseous ozone treatments were tested, including single, double, and triple treatments, with durations ranging from 10 to 60 min and concentrations from 3.3 to 20 ppm. Additionally, ozonated water treatments were evaluated with concentrations of 2, 4, and 6 ppm. To simulate a realistic infestation scenario, mandarins were artificially infected with P. italicum spores before undergoing both gaseous ozone and ozonated water treatments. The storage conditions for the mandarins were meticulously controlled, maintaining a humidity level of 50–60% and a temperature range of 10–12 °C. Each fruit was analyzed, and the presence of P. italicum infection was determined two and three weeks after the ozonation. Results indicated that ozone treatments significantly reduced mold growth, with gaseous ozone demonstrating efficacy rates up to 97.5% and ozonated water treatments achieving preservation rates between 95% and 97%. These results underscore ozone’s potential as a safe, efficient, and sustainable alternative to conventional fungicides, offering promising solutions for extending the shelf life of mandarins. Further research is recommended to optimize ozone treatment parameters, assess long-term effects on fruit quality and nutritional content, and refine application techniques to harness ozone’s potential in citrus fruit preservation fully. This approach not only addresses food security challenges but also aligns with global efforts to reduce chemical inputs in agriculture and promote environmentally sustainable practices. Full article

Penicillium italicum, a major postharvest pathogen, causes blue mold rot in citrus fruits through the deployment of various virulence factors. Recent studies highlight the role of the epigenetic reader, SntB, in modulating the pathogenicity of phytopathogenic fungi. Our research revealed that the deletion of the SntB gene in P. italicum led to significant phenotypic alterations, including delayed mycelial growth, reduced spore production, and decreased utilization of sucrose. Additionally, the mutant strain exhibited increased sensitivity to pH fluctuations and elevated iron and calcium ion stress, culminating in reduced virulence on Gannan Novel oranges. Ultrastructural analyses disclosed notable disruptions in cell membrane integrity, disorganization within the cellular matrix, and signs of autophagy. Transcriptomic data further indicated a pronounced upregulation of hydrolytic enzymes, oxidoreductases, and transport proteins, suggesting a heightened energy demand. The observed phenomena were consistent with a carbon starvation response potentially triggering apoptotic pathways, including iron-dependent cell death. These findings collectively underscored the pivotal role of SntB in maintaining the pathogenic traits of P. italicum, proposing that targeting PiSntB could offer a new avenue for controlling citrus fungal infections and subsequent fruit decay. Full article

This study identified and tested fruit-isolated Metschnikowia yeasts against three major postharvest citrus pathogens, namely, Penicillium digitatum, Penicillium italicum, and Geotrichum citri-aurantii, and further evaluated the impact of FeCl₃ on the biocontrol efficiency of pulcherrimin-producing M. pulcherrima strains. Based on the characterization of the pigmented halo surrounding the colonies and the analysis of the D1/D2 domain of 26S rDNA, a total of 46 Metschnikowia sp. were screened and identified. All 46 Metschnikowia strains significantly inhibited the hyphal growth of Penicillium digitatum, Penicillium italicum, and Geotrichum citri-aurantii, and effectively controlled the development of green mold, blue mold and sour rot of citrus fruit. The introduction of exogenous FeCl₃ at certain concentrations did not significantly impact the pulcherriminic acid (PA) production of pigmented M. pulcherrima strains, but notably diminished the size of pigmented zones and the biocontrol efficacy against the three pathogens. Iron deficiency sensitivity experiments revealed that P. digitatum and P. italicum exhibited higher sensitivity compared to G. citri-aurantii, indicating that iron dependence varied among the three pathogens. These results suggested that M. pulcherrima strains, capable of producing high yields of PA, possessed great potential for use as biocontrol agents against postharvest citrus diseases. The biocontrol efficacy of these yeasts is mainly attributed to their ability to competitively deplete iron ions in a shared environment, with the magnitude of their pigmented halo directly correlating to their antagonistic capability. It is worth noting that the level of sensitivity of pathogens to iron deficiency might also affect the biocontrol effect of pulcherrimin-producing M. pulcherrima. Full article

Citrus fruits are vulnerable to green mold (caused by Penicillium digitatum) and blue mold (caused by Penicillium italicum) during storage, posing significant challenges to the industry. Therefore, biological control utilizing antagonistic bacteria has emerged as a dependable strategy for managing postharvest diseases. In this study, halophilic bacterial isolates were carefully selected from diverse saline ecosystems, including the Dead Sea, the Agadir Sea, the Rabat Sea, saline soil, and water of the Amassine Oued in Taounate, based on rigorous in vitro and in vivo antagonism bioassays. Out of 21 bacteria from different saline environments, 10 were chosen for further characterization based on the 16S rDNA gene. Notably, the EAM1 isolate demonstrated exceptional inhibitory effects, reaching a 90% inhibition rate against P. digitatum, while the ER2 isolate closely followed with an 89% inhibition rate against P. italicum. Furthermore, in bacterial supernatant experiments, six bacterial isolates effectively curbed the growth of P. digitatum, and three demonstrated efficacy against P. italicum development. In an in vivo trial spanning ten days of incubation, three highly effective isolates against P. digitatum displayed zero severity, and two of these isolates also demonstrated zero severity against P. italicum. Interestingly, a comparison of bacterial filtrates revealed that all isolates exhibited a severity level of over 50% against the pathogen causing green rot (P. digitatum), while the severity was lower than 50% for the supernatants of the two isolates used against P. italicum. In conclusion, this study highlights the promising role of halophilic bacteria, specifically Bacillus amyloliquefaciens EAM1 and B. amyloliquefaciens ER2, in controlling postharvest fruit pathogens. The findings shed light on the potential of utilizing these bioprotective agents to address the challenges posed by green and blue citrus molds, providing valuable insights for the citrus industry. Full article

Mexico is among the most important citrus fruit producers in the world. However, during storage, several problems related to fungi can arise. The most common fungal postharvest diseases detected on Citrus limon var Eureka (Italian lime) produced in the Tamaulipas state are green/blue mold (Penicillium spp.), fusarium rot (F. oxysporum, F. solani, F. proliferatum, among others), and anthracnose (Colletotrichum spp.). In this work, we selected yeasts, occurring as the natural epiphytic mycoflora of lemons or from fermented traditional products, to be tested as part of a formulation for protecting stored lemons against fungal diseases. The best-performing yeasts, labeled as LCBG-03 (Meyerozyma guilliermondii), LCBG-30 (Pseudozyma sp.), and LCBG-49 (Saccharomyces cerevisiae), were selected to test their compatibility and biocontrol performance against strains of Penicillium digitatum (AL-38), Fusarium sp. (AL-21), Colletotrichum gloeosporioides (AL-13), and Epicoccum sorghinum (H3A). Based on their in vitro performance regarding the percentage of radial growth inhibition, both applied individually or as two yeasts mixed at equal cellular concentrations, the best combinations (containing M. guilliermondii formulated with either Pseudozyma sp. or S. cerevisiae) were selected with efficacies higher than 95% in both in vitro fungal radial growth rate inhibition and on stored lemon fruits. This work contributes to the search for compatible yeast combinations with the aim to diminish the fungal losses of citrus fruits using biocontrol for citrus postharvest protection. Full article

Overall, 180 yeasts and bacteria isolated from the peel of citrus fruits were screened for their in vitro antagonistic activity against Penicillium digitatum and P. italicum, causative agents of green and blue mold of citrus fruits, respectively. Two yeast and three bacterial isolates were selected for their inhibitory activity on mycelium growth. Based on the phylogenetic analysis of 16S rDNA and ITS rDNA sequences, the yeast isolates were identified as Candida oleophila and Debaryomyces hansenii while the bacterial isolates were identified as Bacillus amyloliquefaciens, B. pumilus and B. subtilis. All five selected isolates significantly reduced the incidence of decay incited by P. digitatum and P. italicum on ‘Valencia’ orange and ‘Eureka’ lemon fruits. Moreover, they were effective in preventing natural infections of green and blue mold of fruits stored at 4 °C. Treatments with antagonistic yeasts and bacteria did not negatively affect the quality and shelf life of fruits. The antagonistic efficacy of the five isolates depended on multiple modes of action, including the ability to form biofilms and produce antifungal lipopeptides, lytic enzymes and volatile compounds. The selected isolates are promising as biocontrol agents of postharvest green and blue molds of citrus fruits. Full article

Calcium (Ca²⁺)/calmodulin-dependent protein kinases (CaMKs) act as a class of crucial elements in Ca²⁺-signal transduction pathways that regulate fungal growth, sporulation, virulence, and environmental stress tolerance. However, little is known about the function of such protein kinase in phytopathogenic Penicillium species. In the present study, a new CaMK gene from the citrus pathogenic fungus P. italicum, designated PiCaMK1, was cloned and functionally characterized by gene knockout and transcriptome analysis. The open reading frame of PiCaMK1 is 1209 bp in full length, which encodes 402 amino acid residues (putative molecular weight ~45.2 KD) with the highest homologous (~96.3%) to the P. expansum CaMK. The knockout mutant ΔPiCaMK1 showed a significant reduction in vegetative growth, conidiation, and virulence (i.e., to induce blue mold decay on citrus fruit). ΔPiCaMK1 was less sensitive to NaCl- or KCl-induced salinity stress and less resistant to mannitol-induced osmotic stress, indicating the functional involvement of PiCaMK1 in such environmental stress tolerance. In contrast, the PiCaMK1-complemented strain ΔPiCaMK1COM can restore all the defective phenotypes. Transcriptome analysis revealed that knockout of PiCaMK1 down-regulated expression of the genes involved in DNA replication and repair, cell cycle, meiosis, pyrimidine and purine metabolisms, and MAPK signaling pathway. Our results suggested the critical role of PiCaMK1 in regulating multiple physical and cellular processes of citrus postharvest pathogen P. italicum, including growth, conidiation, virulence, and environmental stress tolerance. Full article

Blue mold caused by Penicillium italicum is one of the two major postharvest diseases of citrus fruits. The interactions of pathogens with their hosts are complicated, and virulence factors that mediate pathogenicity have not yet been identified. In present study, a prediction pipeline approach based on bioinformatics and transcriptomic data is designed to determine the effector proteins of P. italicum. Three hundred and seventy-five secreted proteins of P. italicum were identified, many of which (29.07%) were enzymes for carbohydrate utilization. Twenty-nine candidates were further analyzed and the expression patterns of 12 randomly selected candidate effector genes were monitored during the early stages of growth on PDA and infection of Navel oranges for validation. Functional analysis of a cell wall integrity-related gene Piwsc1, a core candidate, was performed by gene knockout. The deletion of Piwsc1 resulted in reduced virulence on citrus fruits, as presented by an approximate 57% reduction in the diameter of lesions. In addition, the mycelial growth rate, spore germination rate, and sporulation of ΔPiwsc1 decreased. The findings provide us with new insights to understand the pathogenesis of P. italicum and develop an effective and sustainable control method for blue mold. Full article

Penicillium italicum (P. italicum), a citrus blue mold, is a pathogenic fungus that greatly affects the postharvest quality of citrus fruits with significant economic loss. Our previous research showed that 2-methoxy-1, 4-naphthoquinone (MNQ) inhibited the growth of Penicillium italicum. However, the water dispersibility of MNQ will limit its further application. Herein, we synthesized MNQ-based carbon dots (2−CDs) with better water dispersibility, which showed a potential inhibitory effect on P. italicum (MIC = 2.8 μg/mL) better than that of MNQ (MIC = 5.0 μg/mL). Transcriptomics integrated with metabolomics reveals a total of 601 differentially enriched genes and 270 differentially accumulated metabolites that are co-mapped as disruptive activity on the cell cytoskeleton, glycolysis, and histone methylation. Furthermore, transmission electron microscopy analysis showed normal appearances and intracellular septum of P. italicum after treatment. These findings contribute tofurther understanding of the possible molecular action of 2−CDs. Full article

A major citrus postharvest pathogen, Penicillium italicum (P. italicum), causes substantial economic losses in citrus. In this study, a citral nanoemulsion containing polymethoxylated flavonoids (PMFs), the antimicrobial compounds from citrus, was prepared. The antifungal activity and potential antifungal mechanisms of the nanoemulsion against P. italicum were evaluated. The results showed that the growth of P. italicum was effectively inhibited by the nanoemulsion, with a minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of 62.5 and 250 mg L⁻¹, respectively. The nanoemulsion significantly inhibited spore germination and mycelial growth, and it altered the morphology of P. italicum. In addition, the permeability of the cell membrane increased with increasing nanoemulsion concentrations, as evidenced by a rapid rise in extracellular electric conductivity and stronger red fluorescence from mycelia (propidium iodide staining). Compared with the control, the nanoemulsion treatment induced a decrease in total lipid and ergosterol contents in P. italicum cells by 64.61% and 60.58%, respectively, demonstrating that membrane integrity had been disrupted. The results indicated that the PMFs-loaded nanoemulsion exerted antifungal activity against P. italicum by disrupting cell membrane integrity and permeability; such a nanoemulsion may be used as a potential fungicide substitute for preservation in citrus fruits. Full article

Blue mold caused by Penicillium italicum is one of the most serious postharvest diseases of citrus fruit. The aim of this study was to investigate the inhibitory effect of a novel oligosaccharide ester, 6-O-β-L-mannopyranosyl-3-O-(2-methylbutanoyl)-4-O-(8-methyldecanoyl)-2-O-(4-methyl-hexanoyl) trehalose (MTE-1), against P. italicum. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM), along with transcriptome and proteome analysis also, were conducted to illuminate the underlying mechanism. Results showed that MTE-1 significantly inhibited P. italicum growth in vitro in a dose-dependent manner. Moreover, MTE-1 suppressed the disease development of citrus fruit inoculated with P. italicum. Furthermore, ultrastructure observation, as well as transcriptome and proteome analysis, indicated that MTE-1 treatment damaged the cell wall and plasma membrane in spores and mycelia of P. italicum. In addition, MTE-1 regulated genes or proteins involved in primary metabolism, cell-wall metabolism, and pathogenicity. These results demonstrate that MTE-1 inhibited P. italicum by damaging cell walls and membranes and disrupting normal cellular metabolism. These findings contribute to the understanding of the possible molecular action of MTE-1. Finally, MTE-1 also provides a new natural strategy for controlling diseases in postharvest fruit. Full article