Cinerea Infections Research Articles

Curse or blessing: Growth- and laccase-modulating properties of polyphenols and their oxidized derivatives on Botrytis cinerea

Infection of grapevines with the grey mold pathogen Botrytis cinerea results in severe problems for winemakers worldwide. Browning of wine is caused by the laccase-mediated oxidation of polyphenols. In the last decades, Botrytis management has become increasingly difficult due to the rising number of resistances and the genetic variety of Botrytis strains. During the search for sustainable fungicides, polyphenols showed great potential to inhibit fungal growth. The present study revealed two important aspects regarding the effects of grape-specific polyphenols and their polymerized oxidation products on Botrytis wild strains. On the one hand, laccase-mediated oxidized polyphenols, which resemble the products found in infected grapes, showed the same potential for inhibition of growth and laccase activity, but differed from their native forms. On the other hand, the impact of phenolic compounds on mycelial growth is not correlated to the effect on laccase activity. Instead, mycelial growth and relative specific laccase activity appear to be modulated independently. All phenolic compounds showed not only inhibitory but also inductive effects on fungal growth and/or laccase activity, an observation which is reported for the first time. The simultaneous inhibition of growth and laccase activity demonstrated may serve as a basis for the development of a natural botryticide. Yet, the results showed considerable differences between genetically distinguishable strains, impeding the use of a specific phenolic compound against the genetic variety of wild strains. The present findings might have important implications for future understanding of Botrytis cinerea infections and sustainable Botrytis management including the role of polyphenols.

Essential oil compounds modulate nutritional quality and stress response in Botrytis cinerea-infected grape (Vitis vinifera L. cv. ‘Karaerik’)

Grapes, rich in vitamins and minerals, could offer a healthy alternative. However, their potential contamination by the fungus Botrytis cinerea poses serious health risks, necessitating cautious consumption, particularly among those on restricted diets, to mitigate adverse effects. Therefore, the study investigated the effects of individual and combined application of individual components of essential oils (EOs) such as thymol, eugenol, and 1,8-cineole on various parameters such as antioxidant enzymes, mineral elements, anthocyanins, and phenolic compounds in the treatment of B. cinerea infection in 'Karaerik' grapes. Our study provided important information about physiology and pathogen response, and results indicated that the pathogen-infected groups had higher levels of oxidative stress indicators, such as proline, malondialdehyde (MDA), and hydrogen peroxide (H2O2). On the other hand, EOs elevate the activity of antioxidant enzymes, notably superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), whereas thymol treatments effectively reduced oxidative stress to levels comparable to the control group. Under thymol and eugenol treatments, nitrogen (N), potassium (K), phosphorus (P), calcium (Ca), magnesium (Mg), sulfur (S), zinc (Zn), manganese (Mn), boron were significantly higher than the control groups. Increases were also determined in the concentrations of mineral elements such as (B) and copper (Cu). Anthocyanin profiles were preserved by EO treatments, with combined EO applications yielding higher anthocyanin profiles percentages compared to individual treatments. In our results, there were notable differences in the overall quantities of phenolic compounds, and the combination of EO applications caused higher levels of phenolic compounds, especially in the groups that include the fungal infection. Overall, the study showed how effective EO treatments are at reducing B. cinerea infections and maintaining grape quality, which has significance for long-term disease control plans and maintaining berry quality in viticulture.

Photodynamic Inactivation in agriculture: combating fungal phytopathogens resistant to conventional treatment

Botrytis cinerea is a severe threat in agriculture, as it can infect over 200 different crop species with gray mold affecting food yields and quality. The conventional treatment using fungicides lead to emerging resistance over the past decades. Here, we introduce Photodynamic Inactivation (PDI) as a strategy to combat B. cinerea infections, independent of fungicide resistance. PDI uses photoactive compounds, which upon illumination create reactive oxygen species toxic for killing target organisms. This study focuses on different formulations of sodium–magnesium–chlorophyllin (Chl, food additive E140) as photoactive compound in combination with EDTA disodium salt dihydrate (Na2EDTA) as cell-wall permeabilizer and a surfactant. In an in vitro experiment, three different photosensitizers (PS) with varying Chl and Na2EDTA concentrations were tested against five B. cinerea strains with different resistance mechanisms. We showed that all B. cinerea mycelial spheres of all tested strains were eradicated with concentrations as low as 224 µM Chl and 3.076 mM Na2EDTA (LED illumination with main wavelength of 395 nm, radiant exposure 106 J cm−2). To further test PDI as a Botrytis treatment strategy in agriculture a greenhouse trial was performed on B. cinerea infected bell pepper plants (Capsicum annum L). Two different rates (560 or 1120 g Ha−1) of PS formulation (0.204 M Chl and 1.279 M Na2EDTA) and a combination of PS formulation with 0.05% of the surfactant BRIJ L4 (560 g Ha−1) were applied weekly for 4 weeks by spray application. Foliar lesions, percentage of leaves affected, percentage of leaf area diseased and AUDPC were significantly reduced, while percentage of marketable plants were increased by all treatments compared to a water treated control, however, did not statistically differ from each other. No phytotoxicity was observed in any treatment. These results add to the proposition of employing PDI with the naturally sourced PS Chl in agricultural settings aimed at controlling B. cinerea disease. This approach seems to be effective regardless of the evolving resistance mechanisms observed in response to conventional antifungal treatments.Graphical abstract

Nano-selenium foliar intervention-induced resistance of cucumber to Botrytis cinerea by activating jasmonic acid biosynthesis and regulating phenolic acid and cucurbitacin.

Botrytis cinerea is the primary disease affecting cucumber production. It can be managed by applying pesticides and cultivating disease-resistant cucumber strains. However, challenges, such as drug resistance in pathogenic bacteria and changes in physiological strains, are obstacles in the effective management of B. cinerea. Nano-selenium (Nano-Se) has potential in enhancing crop resistance to biological stress, but the exact mechanism for boosting disease resistance remains unclear. Here, we used metabolomics and transcriptomics to examine how Nano-Se, as an immune activator, induces plant resistance. Compared with the control group, the application of 10.0 mg/L Nano-Se on the cucumber plant's leaf surface resulted in increased levels of chlorophyll, catalase (10.2%), glutathione (326.6%), glutathione peroxidase (52.2%), cucurbitacin (41.40%), and metabolites associated with the phenylpropane synthesis pathway, as well as the total antioxidant capacity (21.3%). Additionally, the expression levels of jasmonic acid (14.8 times) and related synthetic genes, namely LOX (264.1%), LOX4 (224.1%), and AOC2 (309.2%), were up-regulated. A transcription analysis revealed that the CsaV3_4G002860 gene was up-regulated in the KEGG enrichment pathway in response to B. cinerea infection following the 10.0 mg/L Nano-Se treatment. In conclusion, the activation of the phenylpropane biosynthesis and branched-chain fatty acid pathways by Nano-Se promotes the accumulation of jasmonic acid and cucurbitacin in cucumber plants. This enhancement enables the plants to exhibit resistance against B. cinerea infections. Additionally, this study identified a potential candidate gene for cucumber resistance to B. cinerea induced by Nano-Se, thereby laying a theoretical foundation for further research in this area. © 2023 Society of Chemical Industry.

Linker histone H1 modulates defense priming and immunity in plants.

Linker H1 histones play an important role in animal and human pathogenesis, but their function in plant immunity is poorly understood. Here, we analyzed mutants of the three canonical variants of Arabidopsis H1 histones, namely H1.1, H1.2 and H1.3. We observed that double h1.1h1.2 and triple h1.1h1.2h1.3 (3h1) mutants were resistant to Pseudomonas syringae and Botrytis cinerea infections. Transcriptome analysis of 3h1 mutant plants showed H1s play a key role in regulating the expression of early and late defense genes upon pathogen challenge. Moreover, 3h1 mutant plants showed enhanced production of reactive oxygen species and activation of mitogen activated protein kinases upon pathogen-associated molecular pattern (PAMP) treatment. However, 3h1 mutant plants were insensitive to priming with flg22, a well-known bacterial PAMP which induces enhanced resistance in WT plants. The defective defense response in 3h1 upon priming was correlated with altered DNA methylation and reduced global H3K56ac levels. Our data place H1 as a molecular gatekeeper in governing dynamic changes in the chromatin landscape of defense genes during plant pathogen interaction.

Ozone-Induced Biochemical and Molecular Changes in Vitis vinifera Leaves and Responses to Botrytis cinerea Infections.

To investigate how plants cope with multi-stress conditions, we analyzed the biochemical and molecular changes of Vitis vinifera leaves subjected to single or sequential double stresses (infection by Botrytis cinerea (Bc) and ozone (O3, 100 ppb for 3 h) treatment). In Bc+/O3- leaves, the hydrogen peroxide (H2O2) induction (observed at 12 and 24 h from the end of treatment (FET)) triggered a production of ethylene (Et; +35% compared with Bc-/O3- leaves), which was preceded by an increase of salicylic acid (SA; +45%). This result confirms a crosstalk between SA- and Et-related signaling pathways in lesion spread. The ozone induced an early synthesis of Et followed by jasmonic acid (JA) and SA production (about 2-fold higher), where Et and SA signaling triggered reactive oxygen species production by establishing a feedback loop, and JA attenuated this cycle by reducing Et biosynthesis. In Bc+ + O3+ leaves, Et peaked at 6 and 12 h FET, before SA confirmed a crosstalk between Et- and SA-related signaling pathways in lesion propagation. In O3+ + Bc+ leaves, the H2O2 induction triggered an accumulation of JA and Et, demonstrating a synergistic action in the regulation of defence reactions. The divergence in these profiles suggests a rather complex network of events in the transcriptional regulation of genes involved in the systemic acquired resistance.

Quantification of Botrytis cinerea Growth in Arabidopsis thaliana.

Yield losses attributed to plant pathogens pose a serious threat to plant productivity and food security. Botrytis cinerea is one of the most devastating plant pathogens, infecting a wide array of plant species; it has also been established as a model organism to study plant-pathogen interactions. In this context, development of different assays to follow the relative success of B. cinerea infections is required. Here, we describe two methods to quantify B. cinerea development in Arabidopsis thaliana genotypes through measurements of lesion development and quantification of fungal genomic DNA in infected tissues. This provides two independent techniques that are useful in assessing the susceptibility or tolerance of different Arabidopsis genotypes to B. cinerea. Key features Protocol for the propagation of the necrotrophic plant pathogen fungus Botrytis cinerea and spore production. Two methods of Arabidopsis thaliana infection with the pathogen using droplet and spray inoculation. Two readouts, either by measuring lesion size or by the quantification of fungal DNA using quantitative PCR. The two methods are applicable across plant species susceptible the B. cinerea. Graphical overview A simplified overview of the droplet and spray infection methods used for the determination of B. cinerea growth in different Arabidopsis genotypes.

Alleviating Effects of Linalool Fumigation on Botrytis cinerea Infections in Postharvest Tomato Fruits

Gray mold is a disease that often occurs in postharvest tomato fruits, leading to a significant decline in the fruits’ quality. In production, pesticides are mostly used to control gray mold, which negatively affect both the environment and food safety. The purpose of this article is to study the effects of linalool fumigation on controlling gray mold in tomato fruits and to further investigate the mechanism of linalool function, so as to provide technical support and a theoretical basis for the application of linalool in the green control of tomato gray mold. The results of the in vitro experiments showed that linalool fumigation had a strong inhibitory effect on the mycelial growth of Botrytis cinerea and that the fumigation of linalool inhibited the expansion of pathogens on tomato fruits. The disease index of tomato fruits in the linalool treatment was always lower than that of the control within 72 h after inoculation with Botrytis cinerea; at the end of experiment, the disease index of the control reached 100.0, which was only 8.0 with the 30 μL/L linalool treatment. Linalool fumigation increased the antioxidant capacity of the tomato fruits under Botrytis cinerea infection through regulating activities of SOD, POD and CAT as well as the ascorbic acid (AsA) content, which could be responsible for the lower malondialdehyde (MDA) accumulation. Linalool fumigation increased the activities of polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) in tomato fruits, indicating that secondary metabolism was involved in the function of linalool in the regulation of tomato fruit resistance to pathogens. As the main enzymes related to cell structure, polygalacturonase (PG), cellulase (CL) and β-galactosidase (β-GAL) were inhibited by linalool, which could protect cell wall structures from damage, and strengthened the mechanical barrier against pathogen access to fruit flesh.

Screening of Lactiplantibacillus plantarum Strains from Sourdoughs for Biosuppression of Pseudomonas syringae pv. syringae and Botrytis cinerea in Table Grapes.

Grapes (Vitis vinifera L.) are an essential crop for fresh consumption and wine production. Vineyards are attacked by several economically important bacterial and fungal diseases that require regular pesticide treatment. Among them, Pseudomonas syringae pv. syringae (Ps. syringae) and Botrytis cinerea (B. cinerea) infections cause huge economic losses. The fresh fruit market has shifted to functional natural foodstuffs with clear health benefits and a reduced use of chemicals along the production chain. Lactic acid bacteria (LAB) have a biopreservative effect and are applied to ensure food safety in response to consumers' demands. In the present study, the possibilities of using microorganisms with a potential antimicrobial effect against Ps. syringae and B. cinerea in the production of table grapes were investigated. LAB of the genus Lactiplantibacillus can be a natural antagonist of pathogenic bacteria and fungi by releasing lactic acid, acetic acid, ethanol, carbon dioxide and bacteriocins in the medium. The present study focuses on the characterization of nine Lactiplantibacillus plantarum (Lp. plantarum) strains isolated from spontaneously fermented sourdoughs. Species-specific PCR identified the isolated LAB for partial recA gene amplification with an amplicon size of 318 bp. RAPD-PCR analysis showed the intraspecific diversity of the individual strains. Thirteen plantaricin-like peptides (PlnA, PlnB, PlnC, PlnD, PlnEF, PlnG, PlnI, PlnJ, PlnK, PlnN, PlnNC8, PlnS, and PlnW) produced by isolated Lp. plantarum strains were detected by PCR with gene-specific primers. The key features for future industrial applications were their antimicrobial properties. The culture medium and cell-free supernatant (CFS) were used to establish in vitro antimicrobial activities of Lp. plantarum strains against Ps. syringae and B. cinerea, and inhibition of phytopathogen development was observed. The inhibitory effect of the CFS (cell-free supernatant) of all strains was assessed by infecting table grapes with these pathogens in in vivo experiments. Lp. plantarum Q4 showed the most effective suppression of the pathogens both in vitro and in vivo, which indicates its potential use as a biocontrol agent against berry rot and grey rot on grapes, caused by Ps. syringae and B. cinerea.

Molecular dissection of rose and Botrytis cinerea pathosystems affected by ethylene

Botrytis cinerea (B. cinerea) is a necrotrophic pathogen that causes significant growth and postharvest commercial losses in cut roses. Postharvest disease severity is affected by ethylene levels during transport and storage conditions. In this study, we identified the relationship between ethylene, fungal growth, and ethylene inhibitors during B. cinerea infection in cut ‘Pink Beauty’ roses. The results suggest that cut rose susceptibility to gray mold disease is associated with the developmental stage of flowers, especially the level of senescence, which is stimulated by ethylene biosynthesis in petals. The mRNA levels of ethylene biosynthesis genes in petals were related to the severity of gray mold; however, their expression levels decreased when the cut roses were completely macerated by B. cinerea. B. cinerea infections in cut flowers activated ethylene biosynthesis and signaling pathways, leading to the activation of RhERFs in the petals. Ethylene regulated the development of B. cinerea infections in the cut flowers directly by binding to the receptors or indirectly by promoting the ethylene response in the host tissues. The suppression of ethylene responses in petals and inhibition of the fungal histidine kinase receptor of B. cinerea by 1-methylcylopropene (1-MCP) made cut rose flowers resistant to such necrotrophic pathogens. We also established a working model for ethylene binding and plant and fungus actions in the rose-B. cinerea pathosystem. Understanding the relationship between ethylene and gray mold disease in cut roses and the interaction between ethylene inhibitors and B. cinerea will help improve postharvest treatments for reducing B. cinerea damage in cut flowers.

A novel approach to control Botrytis cinerea fungal infections: uptake and biological activity of antifungals encapsulated in nanoparticle based vectors

Botrytis cinerea, responsible for grey mold diseases, is a pathogen with a broad host range, affecting many important agricultural crops, in pre and post harvesting of fruits and vegetables. Commercial fungicides used to control this pathogen are often subjected to photolysis, volatilization, degradation, leaching, and runoff during application. In this context, the use of a delivery system, based on poly (lactic-co-glycolic acid) nanoparticles (PLGA NPs) represents an innovative approach to develop new pesticide formulations to successfully fight B. cinerea infections. In order to study NPs uptake, B. cinerea conidia and mycelium were treated with PLGA NPs loaded with the high fluorescent probe coumarin 6 (Cu6-PLGA NPs) and analyzed under ApoTome fluorescence microscopy. The observations revealed that 50 nm Cu6-PLGA NPs penetrated into B. cinerea conidia and hyphae, as early as 10 min after administration. Pterostilbene, a natural compound, and fluopyram, a synthetic antifungal, were entrapped in PLGA NPs, added to B. cinerea conidia and mycelium, and their antifungal activity was tested. The results revealed that the compounds loaded in NPs exhibited a higher activity against B. cinerea. These results lay the foundations for the use of PLGA NPs as a new strategy in plant pest management.

Antifungal activities of fluoroindoles against the postharvest pathogen Botrytis cinerea: In vitro and in silico approaches

Botrytis cinerea is a common necrotrophic fungal pathogen, leading cause of gray mold diseases in plants and fruit. Several benzimidazoles are used for controlling B. cinerea-associated infection in fruit and vegetables, but benzimidazoles resistance restricts its further uses. Therefore, it is a need for alternative drugs that control B. cinerea. Indoles are multi-faceted compounds and their structural similarities with antifungal benzimidazoles make them a choice for further investigation. Thus, the main objective of the study was to investigate the antifungal potencies of indoles against B. cinerea and to decipher the molecular mechanism involved. We conducted in vitro antifungal assays, fruit assays, and computational studies of interactions between indoles and fungal microtubule polymerase. Of the 16 halogenated indoles examined, 4-fluoroindole, 5-fluoroindole, and 7-fluoroindole (MIC range 2–5 mg/L) were found to be more potent than the fungicides fluconazole and natamycin. Fluoroindoles inhibited or eradicated B. cinerea infections in tangerines and strawberries. Molecular dynamic simulation and density functional theory showed that these fluoroindoles stably interacted with microtubule polymerase. Quantitative structure-activity relationship analyses of halogenated indoles revealed that the presence of a fluoro group in the indole moiety is essential for anti-Botrytis activity. The plausibility of the underlying antifungal mechanism was confirmed by in vitro tubulin polymerization. Collective outcomes of this study indicates that fluoroindoles could be used as alternative fungicidal agents against B. cinerea.

Neisseria Cinerea Bacteremia Secondary to a Retropharyngeal Abscess

Neisseria cinerea is a commensal bacteria of the human oropharynx, not thought to be pathogenic, and is rarely associated with serious infections, including bacteremia. Case reports involving invasive N. cinerea infections are uncommon in the literature. Retropharyngeal abscesses are unusual in adults, and are usually attributable to local trauma.Based on a review of the literature, Neisseria cinerea bacteremia secondary to a retropharyngeal abscess has not been described. We present a unique case of an elderly female without clear predisposing factors for a retropharyngeal abscess, who presented with a N. cinerea bacteremia and was found to have an asymptomatic retropharyngeal abscess.

Improved optical phenotyping of the grape berry surface using light-separation and automated RGB image analysis

Grape resilience towards Botrytis cinerea (B. cinerea) infections (Botrytis bunch rot) is an important concern of breeders and growers. Beside grape bunch architecture, berry surface characteristics like berry bloom (epicuticular wax) as well as thickness and permeability of the berry cuticle represent further promising physical barriers to increase resilience towards Botrytis bunch rot. In previous studies, two efficient sensor-based phenotyping methods were developed to evaluate both berry surface traits fast and objectively: (1) light-separated RGB (red-green-blue) image analysis to determine the distribution of epicuticular wax on the berry surface; and (2) electrical impedance characteristics of the grape berry cuticle based on point measurements. The present proof-of-concept study aiming at the evaluation of light-separated RGB images for both phenotyping applications, phenotyping wax distribution pattern and berry cuticle impedance values. Within the selected grapevine varieties like 'Riesling', 'Sauvignon Blanc' or 'Calardis Blanc' five contributions were achieved: (1) Both phenotyping approaches were fused into one prototypic unified phenotyping method achieving a wax detection accuracy of 98.6 % and a prediction of electrical impedance with an accuracy of 95 %. (2) Both traits are derived using only light-separated images of the grapevine berries. (3) The improved method allows the detection and quantification of additional surface traits of the grape berry surface such as lenticels (punctual lignification) and the berry stem that are also known as being able to affect the grape susceptibility towards Botrytis. (4) The improved image analysis tools are further integrated into a comprehensive workbench allowing end-users, like breeders to combine phenotyping experiments with transparent data management offering valuable services like visualizations, indexing, etc. (5) Annotation work is supported by a sophisticated annotation tool of the image analysis workbench. The usage of light-separated images enables fast and non-invasive phenotyping of different optical berry surface characteristics, which saves time-consuming labor and additionally allows the reuse of the berry samples for subsequent investigations, e.g. Botrytis infection studies.

Botrytis prunorum Associated to Vitis vinifera Blossom Blight in Chile.

Table grapes are highly susceptible to Botrytis cinerea infections during the bloom period. After reaching the flower development stage, B. cinerea remains quiescent until berry ripening or gives rise to blossom blight under specific climate conditions. A research study was conducted on the Chilean Central Valley during the 2018-2019 growing season. Flowers of Vitis vinifera cv. Thompson Seedless were collected and B. cinerea was isolated together to a second and morphologically different species, characterized by white mycelium and low to no sporulation (11.4% of total isolates). Three randomly selected isolates within this population were genetically examined and identified as Botrytis prunorum based on a phylogenetic multilocus approach using partial regions of genes RPB2, HSP60, and G3PDH or NEP1 and NEP2. Pathogenicity tests showed that B. prunorum infects and causes wilting in healthy table grape flowers. B. prunorum isolates were able to infect Thompson Seedless berries, inducing lesions between 13.11 and 41.53% with respect to the lesion diameter generated by B. cinerea B05.10. The fungicide sensitivity was evaluated. The three genetically characterized isolates were sensitive to boscalid and to cyprodinil/fludioxonil mixture with a mean EC50 value of 5.5 µg/ml and 0.065 µg/ml, respectively. However, loss of sensitivity to fenhexamid was determined, with a mean EC50 value of 5.13 µg/ml. Our understanding about blossom blight in V. vinifera has been limited to B. cinerea. Here we associated B. prunorum as a second causal agent of this disease in Chile. This data represents a first approach to the epidemiological characteristics of B. prunorum associated with blossom blight in table grapes.

The Xylanase Inhibitor TAXI-I Increases Plant Resistance to Botrytis cinerea by Inhibiting the BcXyn11a Xylanase Necrotizing Activity.

During host plant infection, pathogens produce a wide array of cell wall degrading enzymes (CWDEs) to break the plant cell wall. Among CWDEs, xylanases are key enzymes in the degradation of xylan, the main component of hemicellulose. Targeted deletion experiments support the direct involvement of the xylanase BcXyn11a in the pathogenesis of Botrytis cinerea. Since the Triticum aestivum xylanase inhibitor-I (TAXI-I) has been shown to inhibit BcXyn11a, we verified if TAXI-I could be exploited to counteract B. cinerea infections. With this aim, we first produced Nicotiana tabacum plants transiently expressing TAXI-I, observing increased resistance to B. cinerea. Subsequently, we transformed Arabidopsis thaliana to express TAXI-I constitutively, and we obtained three transgenic lines exhibiting a variable amount of TAXI-I. The line with the higher level of TAXI-I showed increased resistance to B. cinerea and the absence of necrotic lesions when infiltrated with BcXyn11a. Finally, in a droplet application experiment on wild-type Arabidopsis leaves, TAXI-I prevented the necrotizing activity of BcXyn11a. These results would confirm that the contribution of BcXyn11a to virulence is due to its necrotizing rather than enzymatic activity. In conclusion, our experiments highlight the ability of the TAXI-I xylanase inhibitor to counteract B. cinerea infection presumably by preventing the necrotizing activity of BcXyn11a.

Pretreatment of the Antagonistic Yeast, Debaryomyces hansenii, With Mannitol and Sorbitol Improves Stress Tolerance and Biocontrol Efficacy.

The effect of exogenous mannitol and sorbitol on the viability of the antagonist yeast, Debaryomyces hansenii, when exposed to oxidative and high-temperature stress was determined. Results indicated that both the 0.1 M mannitol (MT) and 0.1 M sorbitol (ST) treatments improved the tolerance of D. hansenii to subsequent oxidative and high-temperature stress. MT or ST cells had a significantly higher level of cell survival, elevated the gene expression of catalase 1 (CAT1) and copper-zinc superoxide dismutase (SOD1), as well as the corresponding enzyme activity. Treated cells also exhibited a lower accumulation of intracellular reactive oxygen species (ROS), and a higher content of intracellular mannitol and sorbitol relative to non-treated, control yeast cells, when exposed to a subsequent oxidative (30 mM H2O2) or heat (40.5°C) stress for 30 min. Additionally, MT and ST yeast exhibited a higher growth rate in kiwifruit wounds, and a greater ability to inhibit postharvest blue mold (Penicillium expansum) and gray mold (Botrytis cinerea) infections. The present study indicates that increased antioxidant response induced by mannitol and sorbitol in D. hansenii can enhance stress tolerance and biocontrol performance.

Loss of proton/calcium exchange 1 results in the activation of plant defense and accelerated senescence in Arabidopsis

Cytosolic Ca2+ increases in response to many stimuli. CAX1 (H+/Ca2+ exchanger 1) maintains calcium homeostasis by transporting calcium from the cytosol to vacuoles. Here, we determined that the cax1 mutant exhibits enhanced resistance against both an avirulent biotrophic pathogen Pst-avrRpm1 (Pseudomonas syringae pv tomato DC3000 avrRpm1), and a necrotrophic pathogen, B. cinerea (Botrytis cinerea). The defense hormone SA (salicylic acid) and phytoalexin scopoletin, which fight against biotrophs and necrotrophs respectively, accumulated more in cax1 than wild-type. Moreover, the cax1 mutant exhibited early senescence after exogenous Ca2+ application. The accelerated senescence in the cax1 mutant was dependent on SID2 (salicylic acid induction deficient 2) but not on NPR1 (nonexpressor of pathogenesis-related genes1). Additionally, the introduction of CAX1 into the cax1 mutant resulted in phenotypes similar to that of wild-type in terms of Ca2+-conditioned senescence and Pst-avrRpm1 and B. cinerea infections. However, disruption of CAX3, the homolog of CAX1, did not produce an obvious phenotype. Moreover, exogenous Ca2+ application on plants resulted in increased resistance to both Pst-avrRpm1 and B. cinerea. Therefore, we conclude that the disruption of CAX1, but not CAX3, causes the activation of pathogen defense mechanisms, probably through the manipulation of calcium homeostasis or other signals.

RNAi-Based Biofungicides as a Promising Next-Generation Strategy for Controlling Devastating Gray Mold Diseases.

Botrytis cinerea is one of the most critical agro-economic phytopathogens and has been reported to cause gray mold disease in more than 1000 plant species. Meanwhile, small interfering RNA (siRNA), which induce RNA interference (RNAi), are involved in both host immunity and pathogen virulence. B. cinerea has been reported to use both siRNA effectors and host RNAi machinery to facilitate the progression of gray mold in host species. Accordingly, RNAi-based biofungicides that use double-stranded RNA (dsRNA) to target essential fungal genes are considered an emerging approach for controlling devastating gray mold diseases. Furthermore, spray-induced gene silencing (SIGS), in which the foliar application of dsRNA is used to silence the pathogen virulence genes, holds great potential as an alternative to host-induced gene silencing (HIGS). Recently, SIGS approaches have attracted research interest, owing to their ability to mitigate both pre- and post-harvest B. cinerea infections. The RNAi-mediated regulation of host immunity and susceptibility in B. cinerea–host interactions are summarized in this review, along with the limitations of the current knowledge of RNAi-based biofungicides, especially regarding SIGS approaches for controlling gray mold diseases under field conditions.

Tetraclinis articulata essential oil reduces Botrytis cinerea infections on tomato

Essential oils (EOs) extracted from medicinal plants gained interest in research due to their potential effectiveness as antimicrobial compounds that can substitute chemical pesticides for treatment of plant derived foods. This study aims to use Tetraclinis articulata EO to control Botrytis cinerea pre- and post-harvest infection on tomato. Tetraclinis articulata EOs extracted at vegetative, flowering and fructification stages were rich in α-pinene (8.10–21.52%), β-caryophyllene (5.17–16.10 %) and caryophyllene oxide (5.57–6.83 %) with the main chemical classes were oxygenated monoterpenes (30.50–50.86 %) and monoterpene hydrocarbons (12.19–28.96 %). The in vitro antifungal activity of EOs against B. cinerea varied with the phenological stage, with the highest activity observed at flowering stage which used subsequently. The activity of α-pinene alone or mixed with bornyl acetate according to their natural proportions at 250 μg mL−1 totally inhibited B. cinerea conidia germination. The pulverization of EO on detached leaflets at 2 mg mL−1 reduced B. cinerea necrosis to 14.50 % in comparison to only infected leaflets 76.00 %. Tetraclinis articulata EO applied as preventive treatment on tomato plants under greenhouse conditions enhanced plants growth and reduced B. cinerea infection to 17.72 % in comparison to infected and non-treated plants 52.11 %. The EO applied at 100 μg mL-1 on tomato fruit reduced infection caused by B. cinerea by 64.01 %. Tetraclinis articulata EO constitutes a promising natural product to reduce B. cinerea infections on tomato in pre- and post-harvest conditions during storage and transport.