Endophytic Fungi Volatile Organic Compounds as Crucial Biocontrol Agents Used for Controlling Fruit and Vegetable Postharvest Diseases.

Fruits and vegetables are an important part of the human diet, but during transportation and storage, microbial pathogens attack and spoil fruits and vegetables, causing huge economic losses to agriculture. Traditionally used chemical fungicides leave chemical residues, leading to environmental pollution and health risks. With the emphasis on food safety, biocontrol agents are attracting more and more attention due to their environmental friendliness. Endophytic fungi are present in plant tissues and do not cause host disease. The volatile organic compounds (VOCs) they produce are used to control postharvest diseases due to their significant antifungal activity, as well as their volatility, safety and environmental protection characteristics. This review provides the concept and characterization of endophytic fungal VOCs, concludes the types of endophytic fungi that release antifungal VOCs and their biological control mechanisms, as well as focuses on the practical applications and the challenges of applying VOCs as fumigants. Endophytic fungal VOCs can be used as emerging biocontrol resources to control postharvest diseases that affect fruits and vegetables.

Inhibitory Effect and Potential Antagonistic Mechanism of Isolated Epiphytic Yeasts against Botrytis cinerea and Alternaria alternata in Postharvest Blueberry Fruits.

This study evaluated the biocontrol effect of isolated epiphytic yeasts (Papiliotrema terrestris, Hanseniaspora uvarum, and Rhodosporidium glutinis) against Botrytis cinerea and Alternaria alternata in blueberry fruits and its possible mechanisms. Our findings indicated that the three tested yeasts exerted a good biocontrol effect on postharvest diseases in blueberry, and that H. uvarum was the most effective. In addition, the three tested yeasts could improve the postharvest storage quality of blueberry fruits to some extent. H. uvarum demonstrated the strongest direct inhibitory effect on pathogens by suppressing spore germination, mycelial growth, and antifungal volatile organic compound (VOC) production. P. terrestris showed the highest extracellular lytic enzymes activities. It also had better adaptation to low temperature in fruit wounds at 4 °C. The biofilm formation capacity was suggested to be the main action mechanism of R. glutinis, which rapidly colonized fruit wounds at 20 °C. Several action mechanisms are employed by the superb biocontrol yeasts, while yeast strains possess distinctive characteristics and have substantially different action mechanisms.

Bacillus sp. alone or combined with salicylic acid inhibited Trichoderma spp. infection on harvested white Hypsizygus marmoreus.

Introduction: White Hypsizygus marmoreus is a popular edible mushroom. It is rich in nutrition and flavor but vulnerable to fungal disease, resulting in nutrient loss and aging. Methods: In this study, the pathogenic fungus Trichoderma spp. BBP-6 and its antagonist Bacillus sp. 1-23 were isolated and identified. The negative effects caused by this pathogen were judged by detecting a series of changes in the infected white H. marmoreus. The effects of Bacillus sp. 1-23 on Trichoderma spp. BBP-6 and the infected white H. marmoreus were detected. The effect of Bacillus sp. 1-23 treatment combined with salicylic acid (SA) was also considered. Results: The results showed that Trichoderma spp. BBP-6 could affect the activities of antioxidant enzymes PAL, POD, CAT, SOD, GR, PPO, and APX to interfere with the stability of the white H. marmoreus antioxidant enzyme system and cause the mushroom severe browning and nutrition loss, as well as general quality deterioration. Bacillus sp. 1-23 could produce chitinase and chitosanase enzymes to inhibit Trichoderma spp. BBP-6 directly. SA reinforced this inhibitory. Bacillus sp. 1-23 alone or combined with SA could help white H. marmoreus from the Trichoderma spp. BBP-6 infection to effectively maintain nutrients, restore and stabilize the antioxidant system, and reduce the production of malondialdehyde, superoxide anion and hydrogen peroxide. Discussion: Thus, such treatments could be considered potential methods to alleviate damage from disease and extend the shelf life of white H. marmoreus.

Inhibitory effects and mechanisms of cinnamaldehyde against Fusarium oxysporum, a serious pathogen in potatoes.

BACKGROUND: Potatoes, a major economic crop, are significantly impacted by Fusarium dry rot, a prevalent postharvest disease. Despite the broad-spectrum antimicrobial properties of cinnamaldehyde, a naturally-derived plant substance, its efficacy against the causal pathogen of potato dry rot (Fusarium oxysporum) and the underlying mechanisms have not been extensively studied. RESULTS: Our study demonstrates that cinnamaldehyde effectively inhibits the growth of Fusarium oxysporum, the pathogen responsible for potato dry rot, and increases its sensitivity to environmental stress factors such as extreme temperatures and high salt stress. Treatment with cinnamaldehyde results in altered fungal mycelium morphology, compromised cell wall stability, and disrupted cell membrane integrity, thereby reducing spore viability. Specifically, it interferes with the cell membrane and cell wall structures of the fungus, potentially disrupting fungal growth by modulating signaling pathways involved in cell wall maintenance, chitin metabolism, and GPI-anchored protein function. Notably, we show that cinnamaldehyde induces a form of regulated cell death in F. oxysporum, which is characterized not as typical apoptosis, as evidenced by Annexin V negative staining. However, the specific cell death type and underlying mechanism still needed to be further explored. CONCLUSION: Cinnamaldehyde, an environmentally friendly plant-based active compound, exhibits strong inhibitory effects on F. oxysporum, indicating its potential use in the prevention and control strategies for potato dry rot. This research contributes to the understanding of novel antifungal mechanisms and offers promising insights into eco-friendly alternatives for managing this economically significant postharvest disease. © 2024 Society of Chemical Industry.

Mining the Penicillium expansum Genome for Virulence Genes: A Functional-Based Approach to Discover Novel Loci Mediating Blue Mold Decay of Apple Fruit.

Blue mold, a postharvest disease of pome fruits, is caused by the filamentous fungus Penicillium expansum. In addition to the economic losses caused by P. expansum, food safety can be compromised, as this pathogen is mycotoxigenic. In this study, forward and reverse genetic approaches were used to identify genes involved in blue mold infection in apple fruits. For this, we generated a random T-DNA insertional mutant library. A total of 448 transformants were generated and screened for the reduced decay phenotype on apples. Of these mutants, six (T-193, T-275, T-434, T-588, T-625, and T-711) were selected for continued studies and five unique genes were identified of interest. In addition, two deletion mutants (Δt-625 and Δt-588) and a knockdown strain (t-434KD) were generated for three loci. Data show that the ∆t-588 mutant phenocopied the T-DNA insertion mutant and had virulence penalties during apple fruit decay. We hypothesize that this locus encodes a glyoxalase due to bioinformatic predictions, thus contributing to reduced colony diameter when grown in methylglyoxal (MG). This work presents novel members of signaling networks and additional genetic factors that regulate fungal virulence in the blue mold fungus during apple fruit decay.

Insights into the Isolation, Identification, and Biological Characterization Analysis of and Novel Control Strategies for Diaporthe passiflorae in Postharvest Passion Fruit.

Postharvest diseases seriously restrict developments in the passion fruit industry. In this study, we aimed to identify the postharvest pathogen affecting passion fruit, investigate its pathogenicity, and explore relevant control methods. The pathogen was isolated from rotting passion fruit and identified using morphological characteristics, ITS sequences, and phylogenetic tree analyses. Additionally, preliminary studies were conducted to assess the biological characteristics of the pathogen and evaluate the efficacy of various treatments for disease control. The fungus on the passion fruit called B4 was identified as Diaporthe passiflorae. Optimal conditions for mycelial growth were observed at 25-30 °C and pH 5-6, with starch as the carbon source and peptone as the nitrogen source. Infection by D. passiflorae accelerated fruit decay, reduced the h° value of the peel, and increased the peel cell membrane permeability when compared to the control. Notably, treatments with appropriate concentrations of ɛ-poly-l-lysine, salicylic acid, and melatonin showed inhibitory effects on the pathogen's growth in vitro and may thus be potential postharvest treatments for controlling brown rot caused by D. passiflorae in passion fruit. The results provide a scientific basis for the development of strategies to control postharvest decay and extend the storage period of passion fruit.

Corrigendum: Identification of volatile organic compounds in extremophilic bacteria and their effective use in biocontrol of postharvest fungal phytopathogens.

[This corrects the article DOI: 10.3389/fmicb.2021.773092.].

Recent Insights into the Use of Antagonistic Yeasts for Sustainable Biomanagement of Postharvest Pathogenic and Mycotoxigenic Fungi in Fruits with Their Prevention Strategies against Mycotoxins.

Fungi-induced postharvest diseases are the leading causes of food loss and waste. In this context, fruit decay can be directly attributed to phytopathogenic and/or mycotoxin-producing fungi. The U.N. Sustainable Development Goals aim to end hunger by 2030 by improving food security, sustainable agriculture, and food production systems. Antagonistic yeasts are one of the methods presented to achieve these goals. Unlike physical and chemical methods, harnessing antagonistic yeasts as a biological method controls the decay caused by fungi and adsorbs and/or degrades mycotoxins sustainably. Therefore, antagonistic yeasts and their antifungal mechanisms have gained importance. Additionally, mycotoxins' biodetoxification is carried out due to the occurrence of mycotoxin-producing fungal species in fruits. Combinations with processes and agents have been investigated to increase antagonistic yeasts' efficiency. Therefore, this review provides a comprehensive summary of studies on preventing phytopathogenic and mycotoxigenic fungi and their mycotoxins in fruits, as well as biocontrolling and biodetoxification mechanisms.

The Exploitation of Microbial Antagonists against Postharvest Plant Pathogens.

Postharvest disease management is vital to increase the quality and productivity of crops. As part of crop disease protection, people used different agrochemicals and agricultural practices to manage postharvest diseases. However, the widespread use of agrochemicals in pest and disease control has detrimental effects on consumer health, the environment, and fruit quality. To date, different approaches are being used to manage postharvest diseases. The use of microorganisms to control postharvest disease is becoming an eco-friendly and environmentally sounds approach. There are many known and reported biocontrol agents, including bacteria, fungi, and actinomycetes. Nevertheless, despite the abundance of publications on biocontrol agents, the use of biocontrol in sustainable agriculture requires substantial research, effective adoption, and comprehension of the interactions between plants, pathogens, and the environment. To accomplish this, this review made an effort to locate and summarize earlier publications on the function of microbial biocontrol agents against postharvest crop diseases. Additionally, this review aims to investigate biocontrol mechanisms, their modes of operation, potential future applications for bioagents, as well as difficulties encountered during the commercialization process.

Electronic nose as a tool for early detection of diseases and quality monitoring in fresh postharvest produce: A comprehensive review.

Postharvest diseases and quality degradation are the major factors causing food losses in the fresh produce supply chain. Hence, detecting diseases and quality deterioration at the asymptomatic stage of produce enables growers to treat the diseases earlier, maintain quality and reduce postharvest food losses. With the emergence of numerous technologies to detect diseases early and monitor the quality of fresh produce, such as polymerase chain reaction, gas chromatography-mass spectrophotometry, and near-infrared spectroscopy, electronic nose (EN) has also gained acknowledgement and popularity in the past decade as a robust and non-invasive analysis tool to detect odor profile and establish volatile biomarkers for metabolomics databases. However, literature reviewing the EN research on the early detection of diseases in produce after harvest is scarce. The fundamental concept of EN working principles (odor sampling, gas detection, and data acquisition method), as well as the application of EN as a whole, are covered in the first section of the review. An in-depth discussion of the application of EN analysis in the early identification of postharvest diseases and quality monitoring is provided in the subsequent sections, which is the key objective of this comprehensive review. The prospect, limitations, and likely future developments of EN in the postharvest sector are further highlighted in the last section.

Research progress of volatile organic compounds produced by plant endophytic bacteria in control of postharvest diseases of fruits and vegetables.

Pathogen infestation results in significant losses of fruits and vegetables during handling, transportation, and storage. The use of synthetic fungicides has been a common measure for controlling plant pathogens. However, their excessive use of chemicals has led to increased environmental pollution, leaving large amounts of chemicals in agricultural products, posing a threat to human and animal health. There is now an increasing amount of research activities to explore safer and more innovative ways to control plant pathogens. In this regard, endophytic bacteria contribute significantly. Endophytic bacteria are ubiquitous in the internal tissues of plants without causing damage or disease to the host. Due to their high volatility and difficulties in residue in fruits and vegetables, volatile organic chemicals (VOCs) produced by endophytic bacteria have received a lot of attention in recent years. VOCs are a potential biofumigant for the effective control of postharvest fruits and vegetables diseases. This review focuses mainly on the recent progress in using endophytic bacteria VOCs to control post-harvest fruits and vegetables disease. This review provides a brief overview of the concept, characteristics, and summarises the types, application effect, and control mechanisms of endophytic bacterial VOCs. The research area that is being developed has great application value in agriculture and living practice.

Microbial antagonists to biologically control postharvest decay and preserve fruit quality.

Postharvest waste due to decay of fruits and vegetables negatively affects food security, while at the same time control of decay and therefore waste can be limited because of consumer concerns about use of synthetic chemicals. Use of antagonistic microorganisms is an eco-friendly technique that represents a promising alternative approach to the use of chemical methods. Understanding the interactions between antagonists and the fruit microbiome will enable the discovery of new methods to reduce postharvest waste. This article reviews different microbial agents, fungi, bacteria and yeasts that could control decay. Recent developments in the use of microorganisms for preserving postharvest fruit quality, formulation of effective antagonists, and the commercialization steps are also discussed. Antagonists control decay through either direct or indirect mechanisms while preserving the appearance, flavor, texture and nutritional value of horticultural products. Microorganisms do not fully control pathogens, and therefore they are usually used with other treatments or have their biocontrol ability modified through genetic manipulations. Despite of these limitations, commercialization of biocontrol products based on antagonists with required stability and biocontrol potential is occurring. Biocontrol of postharvest decay and waste agent is promising technology for fruit and vegetable industries. Further study is necessary to better understand mechanisms and increasing efficiency of this method.

A Coating Based on Bioactive Compounds from Streptomyces spp. and Chitosan Oligomers to Control Botrytis cinerea Preserves the Quality and Improves the Shelf Life of Table Grapes.

Botrytis cinerea is the most harmful postharvest disease of table grapes. Among the strategies that can be envisaged for its control, the use of coatings based on natural products is particularly promising. The study presented herein focuses on the assessment of the antagonistic capacity of two Streptomyces species and their culture filtrates against B. cinerea. Firstly, the secondary metabolites were characterized by gas chromatography-mass spectrometry, with N1-(4-hydroxybutyl)-N3-methylguanidine acetate and 2R,3S-9-[1,3,4-trihydroxy-2-butoxymethyl]guanine acetate as the main compounds produced by S. lavendofoliae DSM 40217; and cyclo(leucyloprolyl) and cyclo(phenylalanylprolyl) as the most abundant chemical species for S. rochei DSM 41729. Subsequently, the capacity of S. lavendofoliae DSM 40217 and S. rochei DSM 41729 to inhibit the growth of the pathogen was tested in dual culture plate assays, finding 85-90% inhibition. In agar dilution tests, their culture filtrates resulted in effective concentration values (EC90) in the 246-3013 µg·mL-1 range. Upon the formation of conjugate complexes with chitosan oligomers (COS) to improve solubility and bioavailability, a synergistic behavior was observed, resulting in lower EC90 values, ranging from 201 to 953 µg·mL-1. Ex situ tests carried out on 'Timpson' and 'Red Globe' table grapes using the conjugate complexes as coatings were found to maintain the turgor of the grapes and delay the appearance of the pathogen by 10-15 days at concentrations in the 750-1000 µg·mL-1 range. Hence, the conjugate complexes of COS and the selected Streptomyces spp. culture filtrates may be put forward as promising protection treatments for the sustainable control of gray mold.

Endophytic species of Induratia from coffee and carqueja plants from Brazil and its potential for the biological control of toxicogenic fungi on coffee beans by means of antimicrobial volatiles.

Several endophytic fungi have been reported to have produced bioactive metabolites. Some of them, including the Induratia species, have the capacity to emit volatile compounds with antimicrobial properties with broad spectrum against human and plant pathogens. The present study aimed to prospect the Induratia species producing volatile organic compounds (VOCs), in carqueja plants used in alternative medicine and coffee plants in Brazil. A total of 11 fungal isolates producing volatile metabolites were obtained by a parallel growth technique, using I. alba 620 as a reference strain. Phylogenetic relationships revealed the presence of at least three distinct species, I. coffeana, I. yucatanensis, and Induratia sp. SPME/GC/MS analyses of the VOCs in the headspace above the mycelium from Induratia species cultured for 10 days on PDA revealed the volatile profile emitted by I. coffeana CCF 572, I. coffeana COAD 2055, I. yucatanensis COAD 2062, and Induratia sp. COAD 2059. Volatile organic compounds produced by I. coffeana isolates presented antimicrobial activity against Aspergillus ochraceus, A. sclerotiorum, A. elegans, A. foetidus, A. flavus, A. tamari, A. tubingensis, A. sydowii, A. niger, A. caespitosus, A. versicolor, and A. expansum, sometimes by decreasing the growth rate or, mainly, by fully inhibiting colony growth. Fifty-eight percent of the target species died after 6 days of exposure to VOCs emitted by I. coffeana CCF 572. In addition, VOCs emitted by the same fungus inhibited the growth in A. ochraceus inoculated into coffee beans, which indicates that plants which have I. coffeana as an endophyte may be protected from attacks by this plant pathogen.

Effects of thymol concentration on postharvest diseases and quality of blueberry fruit.

In this study, we compared the incidence of postharvest disease and the storage potential of blueberry fruit treated with thymol (inoculated with Aspergillus niger) with those in the control fruit during storage for 42 d at 2 °C. Treatment with 10-30 mg/L thymol was found to be more effective than treatment with higher thymol concentrations of 40-50 mg/L in terms of controlling Aspergillus niger-induced decay. In the thymol-treated blueberry fruit peel, the activities of the disease resistance-related enzymes were significantly enhanced. Furthermore, the 20 mg/L thymol-treated blueberry fruit retained the highest firmness, total soluble solids content, and acceptability score, and it also maintained an unimpaired cell wall structure of pericarp quality. Thus, low-dose thymol-treatment could be a suitable biocontrol agent for controlling postharvest disease and prolonging the storage life of blueberry fruit.

Mechanisms and technology of marine oligosaccharides to control postharvest disease of fruits.

Fruit and vegetable postharvest diseases wreak havoc on food supply and the market economy. Meanwhile, the frequent use of synthetic fungicides poses a threat to the environment and public health. Marine oligosaccharides can act as an elicitor of postharvest disease resistance in fruits and also show broad-spectrum antimicrobial activity. Therefore, this review focuses on the mechanism and technology of marine oligosaccharides for postharvest disease control and compares them to horticulture studies to highlight the limitations of existing research and the prospects for future study. It is critical to developing marine oligosaccharides-based products for postharvest disease control.

Current and future trends in the biocontrol of postharvest diseases.

Postharvest diseases of fruits and vegetables cause significant economic losses to producers and marketing firms. Many of these diseases are caused by necrotrophic fungal pathogens that require wounded or injured tissues to establish an infection. Biocontrol of postharvest diseases is an evolving science that has moved from the traditional paradigm of one organism controlling another organism to viewing biocontrol as a system involving the biocontrol agent, the pathogen, the host, the physical environment, and most recently the resident microflora. Thus, the paradigm has shifted from one of simplicity to complexity. The present review provides an overview of how the field of postharvest biocontrol has evolved over the past 40 years, a brief review of the biology of necrotrophic pathogens, the discovery of BCAs, their commercialization, and mechanisms of action. Most importantly, current research on the use of marker-assisted-selection, the fruit microbiome and its relationship to the pathobiome, and the use of double-stranded RNA as a biocontrol strategy is discussed. These latter subjects represent evolving trends in postharvest biocontrol research and suggestions for future research are presented.

Chemical Composition Profiling and Antifungal Activity of Saffron Petal Extract.

Numerous fungal plant pathogens can infect fresh fruits and vegetables during transit and storage conditions. The resulting infections were mainly controlled by synthetic fungicides, but their application has many drawbacks associated with the threatened environment and human health. Therefore, the use of natural plants with antimicrobial potential could be a promising alternative to overcome the side effects of fungicides. In this regard, this study aimed at evaluating the antifungal activity potential of saffron petal extract (SPE) against three mains important fungal pathogens: Rhizopus stolonifer, Penicillium digitatum and Botritys cinerea, which cause rot decay on the tomato, orange and apple fruits, respectively. In addition, the organic composition of SPE was characterized by attenuated total reflection Fourier transform infrared (ATR-FT-IR) spectroscopy and its biochemical, and gas chromatography-mass spectrometry (GC-MS) analyses were carried out. The obtained results highlighted an increased inhibition rate of the mycelial growth and spore germination of the three pathogenic fungi with increasing SPE concentrations. The mycelial growth and spore germination were completely inhibited at 10% of the SPE for Rhizopus stolonifer and Penicillium digitatum and at 5% for B. cinerea. Interestingly, the in vivo test showed the complete suppression of Rhizopus rot by the SPE at 10%, and a significant reduction of the severity of grey mold disease (37.19%) and green mold, when applied at 5 and 10%, respectively. The FT-IR spectra showed characteristic peaks and a variety of functional groups, which confirmed that SPE contains phenolic and flavonoid components. In addition, The average value of the total phenolic content, flavonoid content and half-maximal inhibitory concentration (IC50) were 3.09 ± 0.012 mg GAE/g DW, 0.92 ± 0.004 mg QE/g DW and 235.15 ± 2.12 µg/mL, respectively. A volatile analysis showed that the most dominant component in the saffron petal is 2(5H)-Furanone (92.10%). Taken together, it was concluded that SPE could be used as an alternative to antioxidant and antifungal compounds for the control of postharvest diseases in fruits.