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1.
Article in English | MEDLINE | ID: mdl-39141420

ABSTRACT

In Florida, angular leaf spot, caused by Xanthomonas fragariae, was the only known bacterial disease in strawberry, which is sporadic and affects the foliage and calyx. However, from the 2019-2020 to 2023-2024 Florida strawberry seasons, unusual bacterial-like symptoms were observed in commercial farms, with reports of up to 30 % disease incidence. Typical lesions were water-soaked and angular in early stages that later became necrotic with a circular-ellipsoidal purple halo, and consistently yielded colonies resembling Pseudomonas on culture media. Strains were pathogenic on strawberry, fluorescent, oxidase- and arginine-dihydrolase-negative, elicited a hypersensitive reaction on tobacco, and lacked pectolytic activity. Although phenotypic assays, such as fatty acid methyl profiles and Biolog protocols, placed the strains into the Pseudomonas group, there was a low similarity at the species level. Further analysis using 16S rRNA genes, housekeeping genes, and whole genome sequencing showed that the strains cluster into the Pseudomonas group but do not share more than 95 % average nucleotide identity compared to representative members. Therefore, the genomic and phenotypic analysis confirm that the strains causing bacterial spot in strawberry represent a new plant pathogenic bacterial species for which we propose the name Pseudomonas fragariae sp. nov. with 20-417T (17T=LMG 32456T=DSM 113340 T) as the type strain, in relation to Fragaria×ananassa, the plant species from which the pathogen was first isolated. Future work is needed to assess the epidemiology, cultivar susceptibility, chemical sensitivity, and disease management of this possible new emerging strawberry pathogen.


Subject(s)
Bacterial Typing Techniques , DNA, Bacterial , Fragaria , Phylogeny , Plant Diseases , Plant Leaves , Pseudomonas , RNA, Ribosomal, 16S , Fragaria/microbiology , RNA, Ribosomal, 16S/genetics , Plant Diseases/microbiology , Pseudomonas/genetics , Pseudomonas/isolation & purification , Pseudomonas/classification , DNA, Bacterial/genetics , Plant Leaves/microbiology , Florida , Sequence Analysis, DNA , Whole Genome Sequencing , Fatty Acids , Genes, Essential/genetics
2.
Sensors (Basel) ; 24(15)2024 Jul 25.
Article in English | MEDLINE | ID: mdl-39123884

ABSTRACT

In strawberry cultivation, precise disease management is crucial for maximizing yields and reducing unnecessary fungicide use. Traditional methods for measuring leaf wetness duration (LWD), a critical factor in assessing the risk of fungal diseases such as botrytis fruit rot and anthracnose, have been reliant on sensors with known limitations in accuracy and reliability and difficulties with calibrating. To overcome these limitations, this study introduced an innovative algorithm for leaf wetness detection systems employing high-resolution imaging and deep learning technologies, including convolutional neural networks (CNNs). Implemented at the University of Florida's Plant Science Research and Education Unit (PSREU) in Citra, FL, USA, and expanded to three additional locations across Florida, USA, the system captured and analyzed images of a reference plate to accurately determine the wetness and, consequently, the LWD. The comparison of system outputs with manual observations across diverse environmental conditions demonstrated the enhanced accuracy and reliability of the artificial intelligence-driven approach. By integrating this system into the Strawberry Advisory System (SAS), this study provided an efficient solution to improve disease risk assessment and fungicide application strategies, promising significant economic benefits and sustainability advances in strawberry production.


Subject(s)
Artificial Intelligence , Fragaria , Plant Diseases , Plant Leaves , Fragaria/microbiology , Plant Diseases/microbiology , Neural Networks, Computer , Algorithms , Botrytis
3.
BMC Plant Biol ; 24(1): 692, 2024 Jul 20.
Article in English | MEDLINE | ID: mdl-39030484

ABSTRACT

The bacterial microbiome plays crucial role in plants' resistance to diseases, nutrient uptake and productivity. We examined the microbiome characteristics of healthy and unhealthy strawberry farms, focusing on soil (bulk soil, rhizosphere soil) and plant (roots and shoots). The relative abundance of most abundant taxa were correlated with the chemical soil properties and shoot niche revealed the least amount of significant correlations between the two. While alpha and beta diversities did not show differences between health groups, we identified a number of core taxa (16-59) and marker bacterial taxa for each healthy (Unclassified Tepidisphaerales, Ohtaekwangia, Hydrocarboniphaga) and dysbiotic (Udaeobacter, Solibacter, Unclassified Chitinophagales, Unclassified Nitrosomonadaceae, Nitrospira, Nocardioides, Tardiphaga, Skermanella, Pseudomonas, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Curtobacterium) niche. We also revealed selective pressure of strawberry rhizosphere soil and roots plants in unhealthy plantations increased stochastic ecological processes of bacterial microbiome assembly in shoots. Our findings contribute to understanding sustainable agriculture and plant-microbiome interactions.


Subject(s)
Bacteria , Fragaria , Microbiota , Rhizosphere , Soil Microbiology , Fragaria/microbiology , Bacteria/classification , Bacteria/genetics , Plant Roots/microbiology , Plant Shoots/microbiology , Farms
4.
Pestic Biochem Physiol ; 203: 105995, 2024 Aug.
Article in English | MEDLINE | ID: mdl-39084769

ABSTRACT

Gray mold disease, caused by Botrytis cinerea is a major postharvest disease impacting fruits such as strawberries and tomatoes. This study explores the use of volatile organic compounds (VOCs) produced by Bacillus spp. as eco-friendly biocontrol agents against B. cinerea. In vitro experiments demonstrated that VOCs from Bacillus sp. LNXM12, B. thuringiensis GBAC46, and B. zhanghouensis LLTC93-VOCs inhibited fungal growth by 61.2%, 40.5%, and 21.6%, respectively, compared to the control. LNXM12 was selected for further experiments due to its highest control efficacy of 58.3% and 76.6% on tomato and strawberry fruits, respectively. The LNXM12 VOCs were identified through gas chromatography-mass spectrometry (GC-MS) analysis, and 22 VOCs were identified. Synthetic VOCs with the highest probability percentage, namely ethyloctynol, 3-methyl-2-pentanone (3M2P), 1,3-butadiene-N, N-dimethylformamide (DMF), and squalene were used in experiments. The results showed that the synthetic VOCs ethyloctynol and 3M2P were highly effective, with an inhibition rate of 56.8 and 57.1% against fungal mycelium radial growth at 120 µg/mL on agar plates. Trypan blue staining revealed strongly disrupted, deeper blue, and lysed mycelium in VOC-treated B. cinerea. The scanning and transmission electron microscope (SEM and TEM) results showed that fungal mycelium was smaller, irregular, and shrunken after synthetic VOC treatments. Furthermore, the synthetic VOCs Ethyloctynol and 3M2P revealed high control efficacy on tomatoes and strawberries infected by B. cinerea. The control efficacy on leaves was 67.2%, 66.1% and 64.5%, 78.4% respectively. Similarly, the control efficiency on fruits was 45.5%, 67.3% and 46.3% 65.1%. The expression of virulence genes in B. cinerea was analyzed, and the results revealed that selected genes BcSpl1, BcXyn11A, BcPG2, BcNoxB, BcNoxR, and BcPG1 were downregulated after VOCs treatment. The overall result revealed novel mechanisms by which Bacillus sp. volatiles control postharvest gray mold disease.


Subject(s)
Bacillus , Botrytis , Fragaria , Plant Diseases , Solanum lycopersicum , Volatile Organic Compounds , Botrytis/drug effects , Volatile Organic Compounds/pharmacology , Volatile Organic Compounds/chemistry , Solanum lycopersicum/microbiology , Fragaria/microbiology , Bacillus/drug effects , Plant Diseases/microbiology , Plant Diseases/prevention & control , Antifungal Agents/pharmacology , Gas Chromatography-Mass Spectrometry , Fungicides, Industrial/pharmacology , Biological Control Agents/pharmacology , Fruit/microbiology , Fruit/chemistry
5.
Compr Rev Food Sci Food Saf ; 23(4): e13417, 2024 Jul.
Article in English | MEDLINE | ID: mdl-39072989

ABSTRACT

Strawberries spoil rapidly after harvest due to factors such as the ripening process, weight loss, and, most importantly, microbial contamination. Traditionally, several methods are used to preserve strawberries after harvest and extend their shelf life, including thermal, plasma, radiation, chemical, and biological treatments. Although these methods are effective, they are a concern from the perspective of safety and consumer acceptance of the treated food. To address these issues, more advanced environment-friendly technologies have been developed over the past decades, including modified and controlled atmosphere packaging, active biopolymer-based packaging, or edible coating formulations. This method can not only significantly extend the shelf life of fruit but also solve safety concerns. Some studies have shown that combining two or more of these technologies can significantly extend the shelf life of strawberries, which could significantly contribute to expanding the global supply chain for delicious fruit. Despite the large number of studies underway in this field of research, no systematic review has been published discussing these advances. This review aims to cover important information about postharvest physiology, decay factors, and preservation methods of strawberry fruits. It is a pioneering work that integrates, relates, and discusses all information on the postharvest fate and handling of strawberries in one place. Additionally, commercially used techniques were discussed to provide insight into current developments in strawberry preservation and suggest future research directions in this field of study. This review aims to enrich the knowledge of academic and industrial researchers, scientists, and students on trends and developments in postharvest preservation and packaging of strawberry fruits.


Subject(s)
Food Packaging , Food Preservation , Fragaria , Fruit , Fragaria/microbiology , Fragaria/growth & development , Food Preservation/methods , Food Packaging/methods , Fruit/microbiology , Food Storage/methods
6.
Plant Cell Rep ; 43(8): 201, 2024 Jul 24.
Article in English | MEDLINE | ID: mdl-39048858

ABSTRACT

KEY MESSAGE: Gene silencing of BcDCL genes improves gray mold disease control in the cultivated strawberry. Gene silencing technology offers new opportunities to develop new formulations or new pathogen-resistant plants for reducing impacts of agricultural systems. Recent studies offered the proof of concept that the symptoms of gray mold can be reduced by downregulating Dicer-like 1 (DCL1) and 2 (DCL2) genes of Botrytis cinerea. In this study, we demonstrate that both solutions based on dsRNA topical treatment and in planta expression targeting BcDCL1 and BcDCL2 genes can be used to control the strawberry gray mold, the most harmful disease for different fruit crops. 50, 70 and 100 ng µL-1 of naked BcDCL1/2 dsRNA, sprayed on plants of Fragaria x ananassa cultivar Romina in the greenhouse, displayed significant reduction of susceptibility, compared to the negative controls, but to a lesser extent than the chemical fungicide. Three independent lines of Romina cultivar were confirmed for their stable expression of the hairpin gene construct that targets the Bc-DCL1 and 2 sequences (hp-Bc-DCL1/2), and for the production of hp construct-derived siRNAs, by qRT-PCR and Northern blot analyses. In vitro and in vivo detached leaves, and fruits from the hp-Bc-DCL1/2 lines showed significantly enhanced tolerance to this fungal pathogen compared to the control. This decreased susceptibility was correlated to the reduced fungal biomass and the downregulation of the Bc-DCL1 and 2 genes in B. cinerea. These results confirm the potential of both RNAi-based products and plants for protecting the cultivated strawberry from B. cinerea infection, reducing the impact of chemical pesticides on the environment and the health of consumers.


Subject(s)
Botrytis , Fragaria , Plant Diseases , RNA Interference , Fragaria/genetics , Fragaria/microbiology , Botrytis/pathogenicity , Plant Diseases/microbiology , Plant Diseases/prevention & control , Plant Diseases/genetics , RNA, Double-Stranded/genetics , Gene Expression Regulation, Plant , Plant Proteins/genetics , Plant Proteins/metabolism , Disease Resistance/genetics
7.
Plant Sci ; 347: 112179, 2024 Oct.
Article in English | MEDLINE | ID: mdl-39004407

ABSTRACT

Ethylene response factor (ERF) is a class of plant-specific transcription factors that play an important role in plant growth, development, and stress response. However, the underlying mechanism of strawberry ERFs in pathogenic responses against Botrytis cinerea (B. cinerea) remains largely unclear. In this study, we isolated FaERF2, a nucleus-localized ERF transcription factor from Fragaria x ananassa. Transiently overexpressing FaERF2 in strawberry fruits significantly enhances their resistant ability to B. cinerea, while silencing FaERF2 in strawberry fruits enhances their susceptibility to B. cinerea. In addition, we found that FaERF2 could directly bind to the cis-acting element GCC box in the promoters of two ß-1,3-glucanase genes, FaBG-1 and FaBG-2, and activate their expression. Finally, both strawberry fruits transient expression followed by B. cinerea inoculation assays and recombinant protein incubation tests collectively substantiated the inhibitory effect of FaBG-1 and FaBG-2 on B. cinerea mycelium growth. These results revealed the molecular regulation mechanism of FaERF2 in response to B. cinerea and laid foundations for creating disease-resistance strawberry cultivar through genome editing approach.


Subject(s)
Botrytis , Disease Resistance , Fragaria , Plant Diseases , Plant Proteins , Botrytis/physiology , Fragaria/genetics , Fragaria/microbiology , Plant Diseases/microbiology , Plant Diseases/genetics , Disease Resistance/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Gene Expression Regulation, Plant , Glucan 1,3-beta-Glucosidase/metabolism , Glucan 1,3-beta-Glucosidase/genetics
8.
J Agric Food Chem ; 72(32): 18027-18044, 2024 Aug 14.
Article in English | MEDLINE | ID: mdl-39078084

ABSTRACT

Botrytis cinerea (B. cinerea) and Colletotrichum gloeosporioides (C. gloeosporioides) were isolated from the decaying strawberry tissue. The antifungal properties of Monarda didyma essential oil (MEO) and its nanoemulsion were confirmed, demonstrating complete inhibition of the pathogens at concentrations of 0.45 µL/mL (0.37 mg/mL) and 10 µL/mL, respectively. Thymol, a primary component of MEO, was determined as an antimicrobial agent with IC50 values of 34.51 (B. cinerea) and 53.40 (C. gloeosporioides) µg/mL. Hippophae rhamnoides oil (HEO) was confirmed as a potent antioxidant, leading to the development of a thymol-HEO-chitosan film designed to act as an antistaling agent. The disease index and weight loss rate can be reduced by 90 and 60%, respectively, with nutrients also being well-preserved, offering an innovative approach to preservative development. Studies on the antifungal mechanism revealed that thymol could bind to FKS1 to disrupt the cell wall, causing the collapse of mitochondrial membrane potential and a burst of reactive oxygen species.


Subject(s)
Botrytis , Colletotrichum , Food Preservation , Fragaria , Fragaria/chemistry , Fragaria/microbiology , Botrytis/drug effects , Botrytis/growth & development , Colletotrichum/drug effects , Food Preservation/methods , Apoptosis/drug effects , Plant Diseases/microbiology , Food Preservatives/pharmacology , Food Preservatives/chemistry , Oils, Volatile/pharmacology , Oils, Volatile/chemistry , Antifungal Agents/pharmacology , Antifungal Agents/chemistry , Fungicides, Industrial/pharmacology , Fungicides, Industrial/chemistry , Fruit/chemistry , Fruit/microbiology , Thymol/pharmacology , Thymol/chemistry , Surface Properties , Chitosan/chemistry , Chitosan/pharmacology
9.
Appl Microbiol Biotechnol ; 108(1): 398, 2024 Jun 28.
Article in English | MEDLINE | ID: mdl-38940906

ABSTRACT

Grey mould caused by Botrytis cinerea is a devastating disease responsible for large losses to agricultural production, and B. cinerea is a necrotrophic model fungal plant pathogen. Membrane proteins are important targets of fungicides and hotspots in the research and development of fungicide products. Wuyiencin affects the permeability and pathogenicity of B. cinerea, parallel reaction monitoring revealed the association of membrane protein Bcsdr2, and the bacteriostatic mechanism of wuyiencin was elucidated. In the present work, we generated and characterised ΔBcsdr2 deletion and complemented mutant B. cinerea strains. The ΔBcsdr2 deletion mutants exhibited biofilm loss and dissolution, and their functional activity was illustrated by reduced necrotic colonisation on strawberry and grape fruits. Targeted deletion of Bcsdr2 also blocked several phenotypic defects in aspects of mycelial growth, conidiation and virulence. All phenotypic defects were restored by targeted gene complementation. The roles of Bcsdr2 in biofilms and pathogenicity were also supported by quantitative real-time RT-PCR results showing that phosphatidylserine decarboxylase synthesis gene Bcpsd and chitin synthase gene BcCHSV II were downregulated in the early stages of infection for the ΔBcsdr2 strain. The results suggest that Bcsdr2 plays important roles in regulating various cellular processes in B. cinerea. KEY POINTS: • The mechanism of wuyiencin inhibits B. cinerea is closely associated with membrane proteins. • Wuyiencin can downregulate the expression of the membrane protein Bcsdr2 in B. cinerea. • Bcsdr2 is involved in regulating B. cinerea virulence, growth and development.


Subject(s)
Biofilms , Botrytis , Fragaria , Fungal Proteins , Hyphae , Membrane Proteins , Plant Diseases , Botrytis/pathogenicity , Botrytis/genetics , Botrytis/growth & development , Botrytis/drug effects , Biofilms/growth & development , Biofilms/drug effects , Virulence , Hyphae/growth & development , Hyphae/drug effects , Plant Diseases/microbiology , Fragaria/microbiology , Fungal Proteins/genetics , Fungal Proteins/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Vitis/microbiology , Spores, Fungal/growth & development , Spores, Fungal/drug effects , Spores, Fungal/genetics , Gene Deletion
10.
Int J Biol Macromol ; 272(Pt 1): 132834, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38838885

ABSTRACT

The development of novel packaging materials with antimicrobial properties is crucial in preventing the microbial-induced spoilage of fruits, vegetables, and foodborne illnesses. In this study, homojunction g-C3N4 (HCN) photocatalysts with excellent photocatalytic performance were incorporated into a matrix consisting of pullulan/chitosan (Pul/CS). These photocatalysts were then electrostatically spun onto polylactic acid (PLA) films to fabricate PLA@Pul/CS/HCN nanofibrous composite films. The design of the bilayer films aimed to combine the physical properties of PLA film with the excellent antibacterial properties of nanofiber films, thereby achieving synergistic advantages. The incorporation of the HCN photocatalysts resulted in enhanced hydrophobicity, barrier function, and mechanical properties of the composite films. Under visible light irradiation, the PLA@Pul/CS/HCN films exhibited approximately 3.43 log and 3.11 log reductions of Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA), respectively, within 2 h. The excellent antimicrobial performance could be attributed to the synergistic effect of CS and the release of reactive oxygen species (ROS) from HCN. Moreover, the strawberries packaged in the PLA@Pul/CS/HCN film demonstrated diminished quality degradation and a prolonged shelf life following visible light irradiation treatment. This study will provide new insights into the exploration of safe and efficient antimicrobial food packaging.


Subject(s)
Chitosan , Food Packaging , Fruit , Glucans , Light , Polyesters , Glucans/chemistry , Glucans/pharmacology , Polyesters/chemistry , Chitosan/chemistry , Chitosan/pharmacology , Fruit/chemistry , Food Packaging/methods , Food Preservation/methods , Escherichia coli/drug effects , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Reactive Oxygen Species/metabolism , Methicillin-Resistant Staphylococcus aureus/drug effects , Fragaria/microbiology , Nanofibers/chemistry , Microbial Sensitivity Tests , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Graphite , Nitrogen Compounds
11.
Int J Biol Macromol ; 273(Pt 2): 133108, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38876246

ABSTRACT

In this study, propolis was first loaded into a conventional oil-in-water emulsion, which was combined with a chitosan film-forming solution to produce propolis emulsion-loaded film (PEF). Strawberries inoculated with Botrytis cinerea coated with PEF and blank emulsion-loaded films (BEF) were stored for 14 days at 4 °C. Compared to BEF, PEF showed superior mechanical and oxygen barrier properties, as well as antioxidant activities, but higher moisture permeability. PEF showed less oil agglomeration on the film surface after drying, as demonstrated by scanning electron microscopy (SEM) analysis. Compared to uncoated strawberries, coatings did not have a significant effect on weight loss or firmness during storage. In contrast, coated strawberries showed elevated total phenolics, anthocyanins, and ascorbic acid retention; however, PEF-coating yielded higher values. Moreover, the PEF coating resulted in a significantly lower reduction of organic acid and total soluble solids. Mold growth was visible in both uncoated and BEF-coated strawberries after 7 days of storage, while PEF-coated fruits showed no visible mold until the end of storage. Starting from day 4, PEF-coated fruits showed lower mold counts (~2 log CFU/g) than other samples. Therefore, the PEF prepared in this study has application potential for the preservation of fresh fruits.


Subject(s)
Antioxidants , Chitosan , Edible Films , Emulsions , Food Preservation , Fragaria , Propolis , Chitosan/chemistry , Fragaria/microbiology , Fragaria/chemistry , Emulsions/chemistry , Propolis/chemistry , Propolis/pharmacology , Antioxidants/chemistry , Antioxidants/pharmacology , Food Preservation/methods , Food Storage/methods , Fruit/chemistry , Fruit/microbiology , Permeability , Anthocyanins/chemistry , Phenols/chemistry , Botrytis/drug effects
12.
Int J Biol Macromol ; 274(Pt 2): 133488, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38944092

ABSTRACT

Lignin, renowned for its renewable, biocompatible, and environmentally benign characteristics, holds immense potential as a sustainable feedstock for agrochemical formulations. In this study, raw dealkaline lignin (DAL) underwent a purification process involving two sequential solvent extractions. Subsequently, an enzyme-responsive nanodelivery system (Pyr@DAL-NPs), was fabricated through the solvent self-assembly method, with pyraclostrobin (Pyr) loaded into lignin nanoparticles. The Pyr@DAL-NPs shown an average particle size of 250.4 nm, demonstrating a remarkable loading capacity of up to 54.70 % and an encapsulation efficiency of 86.15 %. Notably, in the presence of cellulase and pectinase at a concentration of 2 mg/mL, the release of Pyr from the Pyr@DAL-NPs reached 92.66 % within 120 h. Furthermore, the photostability of Pyr@DAL-NPs was significantly improved, revealing a 2.92-fold enhancement compared to the commercially available fungicide suspension (Pyr SC). Bioassay results exhibited that the Pyr@DAL-NPs revealed superior fungicidal activity against Botrytis cinerea over Pyr SC, with an EC50 value of 0.951 mg/L. Additionally, biosafety assessments indicated that the Pyr@DAL-NPs effectively declined the acute toxicity of Pyr towards zebrafish and posed no negative effects on the healthy growth of strawberry plants. In conclusion, this study presents a viable and promising strategy for developing environmentally friendly controlled-release systems for pesticides, offering the unique properties of lignin.


Subject(s)
Botrytis , Fragaria , Green Chemistry Technology , Lignin , Nanoparticles , Plant Diseases , Strobilurins , Strobilurins/chemistry , Strobilurins/pharmacology , Botrytis/drug effects , Fragaria/microbiology , Nanoparticles/chemistry , Lignin/chemistry , Plant Diseases/microbiology , Plant Diseases/prevention & control , Animals , Fungicides, Industrial/pharmacology , Fungicides, Industrial/chemistry , Zebrafish , Particle Size , Drug Carriers/chemistry
13.
Int J Biol Macromol ; 274(Pt 1): 133273, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38906346

ABSTRACT

Edible coatings, formulated with sodium alginate and various strains of lactic acid bacteria, were evaluated for their effectiveness in extending the shelf life and mitigating microbial risks associated with strawberries. This study specifically employed strains of Lacticaseibacillus paracasei, Lacticaseibacillus rhamnosus, and Lacticaseibacillus plantarum as antimicrobial agents. Through physicochemical property analysis, the alginate-based antimicrobial coating proved most effective in reducing the strawberry weight loss rate, decay index, and ascorbic acid degradation. Over time, all treatments exhibited increased fungal growth. However, strawberries treated with alginate and lactic acid bacteria recorded lower final colony formation counts-6.82 log CFU/g for SA + LPC, 6.04 log CFU/g for SA + LGG, and 6.26 log CFU/g for SA + LP-compared to 8.73 log CFU/g in the control group. In terms of bacterial resistance under gastrointestinal conditions, L. paracasei demonstrated the highest survival rate post-simulated gastric fluid exposure, while L. plantarum showed the greatest resilience post-simulated intestinal fluid exposure. These findings underscore the efficacy of alginate-based antimicrobial coatings in not only enhancing the storage quality of strawberries but also ensuring microbial safety and potential benefits for gut health.


Subject(s)
Alginates , Food Preservation , Fragaria , Fragaria/microbiology , Fragaria/drug effects , Alginates/chemistry , Alginates/pharmacology , Food Preservation/methods , Lactobacillales , Edible Films , Food Storage/methods , Food Microbiology
14.
Plant Genome ; 17(2): e20477, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38822520

ABSTRACT

Aggressive strains of Neopestalotiopsis sp. have recently emerged as devastating pathogens of strawberry (Fragaria × ananassa Duchesne ex Rozier), infecting nearly all plant parts and causing severe outbreaks of leaf spot and fruit rot in Florida and globally. The development of host resistance is imperative due to the absence of fungicides that effectively inhibit Neopestalotiopsis sp. growth on an infected strawberry crop. Here, we analyzed 1578 individuals from the University of Florida's (UF) strawberry breeding program to identify and dissect genetic variation for resistance to Neopestalotiopsis sp. and to explore the feasibility of genomic selection. We found that less than 12% of elite UF germplasm exhibited resistance, with narrow-sense heritability estimates ranging from 0.28 to 0.69. Through genome-wide association studies (GWAS), we identified two loci accounting for 7%-16% of phenotypic variance across four trials and 3 years. Several candidate genes encoding pattern recognition receptors, intra-cellular nucleotide-binding leucine-rich repeats, and downstream components of plant defense pathways co-localized with the Neopestalotiopsis sp. resistance loci. Interestingly, favorable alleles at the largest-effect locus were rare in elite UF material and had previously been unintentionally introduced from an exotic cultivar. The array-based markers and candidate genes described herein provide the foundation for targeting this locus through marker-assisted selection. The predictive abilities of genomic selection models, with and without explicitly modeling peak GWAS markers as fixed effects, ranged between 0.25 and 0.59, suggesting that genomic selection holds promise for enhancing resistance to Neopestalotiopsis sp. in strawberry.


Subject(s)
Disease Resistance , Fragaria , Genome-Wide Association Study , Plant Diseases , Fragaria/genetics , Fragaria/microbiology , Plant Diseases/microbiology , Plant Diseases/genetics , Disease Resistance/genetics , Ascomycota/pathogenicity , Ascomycota/physiology
15.
Carbohydr Polym ; 339: 122238, 2024 Sep 01.
Article in English | MEDLINE | ID: mdl-38823908

ABSTRACT

The study aimed to develop a novel, transparent and non-toxic coating with antimicrobial, antioxidant, and antifogging properties. The p-coumaric acid-grafted chitosan (CS-PCA) was synthesized via a carbodiimide coupling reaction and then characterized. The CS-PCA coatings were further prepared using the casting method. The CS-PCA coatings obtained exhibited excellent transparency, UV-light barrier ability, and antifogging properties, as confirmed by spectroscopy and antifogging tests. The CS-PCA coatings showed stronger antioxidant capacity and antimicrobial properties against Escherichia coli, Staphylococcus aureus and Botrytis cinerea compared to CS. The multifunctional coatings were further coated on the polyethylene cling film and their effectiveness was confirmed through a strawberry preservation test. The decay of the strawberries was reduced by CS-PCA coated film at room temperature.


Subject(s)
Antioxidants , Chitosan , Coumaric Acids , Escherichia coli , Food Packaging , Fragaria , Fruit , Propionates , Staphylococcus aureus , Chitosan/chemistry , Chitosan/pharmacology , Coumaric Acids/chemistry , Coumaric Acids/pharmacology , Antioxidants/pharmacology , Antioxidants/chemistry , Escherichia coli/drug effects , Staphylococcus aureus/drug effects , Fragaria/microbiology , Food Packaging/methods , Fruit/chemistry , Propionates/chemistry , Propionates/pharmacology , Botrytis/drug effects , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Microbial Sensitivity Tests
16.
Food Microbiol ; 122: 104544, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38839230

ABSTRACT

The objective of this study was to identify a suitable surrogate for E. coli O157:H7 strain 19685/91 and O113:H21 strain TS18/08, by assessing their thermal resistance at temperatures of 60 °C, 65 °C, and 72 °C in strawberry nectar. The influence of the matrix and the research methodology on the decimal reduction time (D-value) was investigated. Thermal kinetics and safety assessment demonstrated that E. coli ATCC 8739 is a suitable surrogate. The study demonstrated that the presence of fruit particles in the nectar increased thermal resistance of the tested strains. Variations in D-values were observed depending on the research method employed, with D-values in glass capillaries were up to 6.6 times lower compared to larger sample volumes. Encapsulation of E. coli ATCC 8739 exhibited high efficiency of 90.25 ± 0.26% and maintained stable viable counts after 26 days of storage in strawberry nectar at 4 °C. There were no significant differences in thermal resistance between surrogates directly inoculated into strawberry nectar and those encapsulated in alginate beads. Additionally, the encapsulated strains did not migrate outside the beads. Therefore, encapsulated E. coli ATCC 8739 in alginate beads can be effectively utilized in industrial settings to validate thermal treatments as a reliable and safe method.


Subject(s)
Enterohemorrhagic Escherichia coli , Fragaria , Fruit , Hot Temperature , Fruit/microbiology , Fragaria/microbiology , Enterohemorrhagic Escherichia coli/growth & development , Food Microbiology , Colony Count, Microbial , Microbial Viability , Plant Nectar/chemistry , Escherichia coli O157/growth & development , Food Contamination/analysis , Food Contamination/prevention & control , Kinetics
17.
Virus Genes ; 60(4): 402-411, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38717669

ABSTRACT

A wide diversity of mycoviruses has been reported from Botrytis species, some with the potential to suppress the pathogenic abilities of this fungus. Considering their importance, this study was devised to find potential hypovirulence-associated mycoviruses found in Botrytis cinerea strains isolated from Pakistani strawberry fields. Here we report the complete genome characterization of two fusariviruses co-infecting a single isolate of phytopathogenic fungus B. cinerea (Kst14a). The viral genomes were sequenced by deep sequencing using total RNA fractions of the Kst14a isolate. The identified viruses were tentatively named Botrytis cinerea fusarivirus 9 (BcFV9) and Botrytis cinerea fusarivirus 3a (BcFV3a). Both viruses had a single-segmented (ssRNA) genome having a size of 6424 and 8370 nucleotides encoding two discontinuous open reading frames (ORFs). ORF-1 of both mycoviruses encodes for a polyprotein having a conserved domain of RNA-dependent RNA polymerase (RdRP) and a helicase domain (Hel) which function in RNA replication, while ORF2 encodes a hypothetical protein with an unknown function, respectively. Phylogenetic analysis indicated that BcFV9 made a clade with the genus Alphafusarivirus and BcFV3a fall in the genus Betafusarivirus in the family Fusariviridae. To our knowledge, this is the first report of two fusariviruses identified in isolates of B. cinerea from Pakistan. Both mycoviruses successfully transfected to a compatible strain of B. cinerea (Mst11). A comparison of virus-free (VF) and virus-infected (VI) isogenic lines showed the presence of these viruses was causing hypovirulence in infected strains. Virus-infected strains also had a small lesion size while testing the pathogenicity via apple assay.


Subject(s)
Botrytis , Fungal Viruses , Genome, Viral , Open Reading Frames , Phylogeny , Plant Diseases , Botrytis/virology , Botrytis/genetics , Fungal Viruses/genetics , Fungal Viruses/isolation & purification , Fungal Viruses/classification , Plant Diseases/microbiology , RNA, Viral/genetics , Fragaria/microbiology , Fragaria/virology , Pakistan , Viral Proteins/genetics , High-Throughput Nucleotide Sequencing
18.
J Agric Food Chem ; 72(20): 11392-11404, 2024 May 22.
Article in English | MEDLINE | ID: mdl-38717972

ABSTRACT

Methylesterases (MESs) hydrolyze carboxylic ester and are important for plant metabolism and defense. However, the understanding of MES' role in strawberries against pathogens remains limited. This study identified 15 FvMESs with a conserved catalytic triad from the Fragaria vesca genome. Spatiotemporal expression data demonstrated the upregulated expression of FvMESs in roots and developing fruits, suggesting growth involvement. The FvMES promoter regions harbored numerous stress-related cis-acting elements and transcription factors associated with plant defense mechanisms. Moreover, FvMES2 exhibited a significant response to Botrytis cinerea stress and showed a remarkable correlation with the salicylic acid (SA) signaling pathway. Molecular docking showed an efficient binding potential between FvMES2 and methyl salicylate (MeSA). The role of FvMES2 in MeSA demethylation to produce SA was further confirmed through in vitro and in vivo assays. After MeSA was applied, the transient overexpression of FvMES2 in strawberries enhanced their resistance to B. cinerea compared to wild-type plants.


Subject(s)
Botrytis , Fragaria , Plant Proteins , Salicylates , Disease Resistance/genetics , Fragaria/enzymology , Fragaria/genetics , Fragaria/microbiology , Fruit/enzymology , Fruit/genetics , Fruit/microbiology , Gene Expression Regulation, Plant , Molecular Docking Simulation , Multigene Family , Plant Diseases/immunology , Plant Diseases/microbiology , Plant Proteins/genetics , Plant Proteins/metabolism , Salicylates/metabolism
19.
Braz J Biol ; 84: e276874, 2024.
Article in English | MEDLINE | ID: mdl-38808783

ABSTRACT

Strawberry (Fragaria x ananassa Duch.) is a highly perishable fruit whose characteristics make it susceptible to developing microorganisms. Plant extracts have been studied as an alternative to pesticides to control spoilage microorganisms, responding to the expectation of the population seeking a healthier way of life. The fungus Botrytis cinerea is a facultative pathogen of vegetables, which can affect all stages of the development of several fruits, such as the strawberry, where it causes gray rot. Trichilia catigua (catuaba), Paullinia cupana (guarana), Stryphnodendron barbatiman (barbatimão), and Caesalpinia peltophoroides (sibipiruna) are planted in the Brazilian flora and have demonstrated pharmacological properties in their extracts. This work aimed to treat strawberries with a biodegradable film containing extracts of these species to evaluate strawberry conservation. There were notable distinctions in mass loss between the extract-treated and control samples. The pH, total acidity (TA), and soluble solids parameters exhibited consistently significant means across both sets of samples. Luminosity increased over the course of days in the color parameters, with the exception of strawberries coated with guarana. The red color showed greater intensity, except for those coated with barbatimão extract. Considering the results, it is possible to conclude that the coatings used can become an alternative to enhance the conservation of strawberries.


Subject(s)
Fragaria , Plant Extracts , Fragaria/chemistry , Fragaria/microbiology , Plant Extracts/pharmacology , Plant Extracts/chemistry , Botrytis/drug effects , Paullinia/chemistry , Caesalpinia/chemistry , Fruit/chemistry
20.
J Sci Food Agric ; 104(11): 6756-6768, 2024 Aug 30.
Article in English | MEDLINE | ID: mdl-38563620

ABSTRACT

BACKGROUND: Most microorganisms that cause food decay and the lower the shelf life of foods are fungi. Nanotechnologies can combat various diseases and deal with the application of nanomaterial to target cells or tissues. In this study selenium nanoparticles (Se-NPs) were synthesized using ascorbic acid and characterized by ultraviolet-visible spectroscopy, transmission electron microscopy (TEM), X-ray diffraction and zeta potential. The different concentrations of As/Se-NPs were tested against various fungi, including Alternaria linicola, Alternaria padwickii, Botrytis cinerea, Bipolaris sp., Cephalosporium acremonium, Fusarium moniliform and Fusarium semitectum. This study tested the influence of coated As/Se-NPs on healthy strawberry fruits and those infected with Botrytis cinerea during 16 days of storage, with regard to shelf life, decay percentage, weight loss, total titratable acidity percentage, total soluble solids content (TSS) and anthocyanin content. RESULTS: Energy-dispersive X-ray analysis showed only two elements: selenium and oxygen. TEM images showed that the nanoparticles ranged in size between 26 to 39 nm and were rhombohedral in shape. Se-NPs showed antifungal activity against all tested fungi, the most effective being against Botrytis cinerea, Cephalosporium acremonium and Fusarium semitectum. During storage periods of strawberries fruits coated with As/Se-NPs, the shelf life was increased, and the number of decaying fruits was less than in control (uncoated) and coated infected fruits. The decline in weight loss was lower in coated fruits than in control fruits. CONCLUSION: These findings demonstrated that As/Se-NPs could effectively maintain the postharvest quality of strawberries, even when the fruit was infected with B. cinerea. © 2024 Society of Chemical Industry.


Subject(s)
Botrytis , Food Preservation , Fragaria , Fruit , Fungi , Fusarium , Nanoparticles , Plant Diseases , Selenium , Fragaria/microbiology , Fragaria/chemistry , Fruit/chemistry , Fruit/microbiology , Food Preservation/methods , Selenium/pharmacology , Selenium/chemistry , Botrytis/drug effects , Botrytis/growth & development , Plant Diseases/microbiology , Plant Diseases/prevention & control , Nanoparticles/chemistry , Fungi/drug effects , Fusarium/drug effects , Fusarium/growth & development , Antifungal Agents/pharmacology , Antifungal Agents/chemistry , Alternaria/drug effects , Alternaria/growth & development , Fungicides, Industrial/pharmacology , Fungicides, Industrial/chemistry , Food Storage
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