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Gray mold and brown rot, caused respectively by Botrytis cinerea and Monilinia spp., are fungal diseases responsible for significant losses during the storage of fruit and vegetables. Nowadays, the control of postharvest diseases is shifting towards more sustainable strategies, including the use of plant secondary metabolites. In this study, the antifungal activity of Origanum vulgare , Thymus vulgaris , Thymus serpyllum , Melaleuca alternifolia , Lavandula officinalis , Lavandula hybrida , Citrus bergamia , Rosmarinus officinalis , Cinnamomum zeylanicum essential oils (EOs) in vapor phase was tested in vitro against B. cinerea, Monilinia fructicola , Monilinia fructigena , and Monilinia laxa . For the experiments, a protocol using a volatile organic compounds (VOC) chamber was designed. Results indicate a dose-dependent inhibitory activity of all the tested EOs, with O. vulgare , T. vulgaris , and T. serpyllum being the most active ones, with minimum inhibitory concentrations (MIC) of 22.73, 45.45, and 22.73 µl/L, respectively, against B. cinerea and a range between 5.64 and 22.73 µl/L against the three Monilinia spp. Overall, B. cinerea presented lower sensitivity to vapor-phase EOs than any of the Monilinia strains, except for the C. zeylanicum EO, which consistently showed higher inhibition against B. cinerea . Among the three Monilinia spp., M. fructicola was the least sensitive, while M. fructigena was the most sensitive. The use of VOC chambers proved to be a reliable protocol for the assessment of antimicrobial activities of EOs. These results suggest that the VOC emitted by the tested EOs are effective towards important decay-causing fungi, and that they could be used for the control of gray mold and brown rot in in vivo trials.

Background Brown rots are important fungal diseases of stone and pome fruits. They are caused by several Monilinia species but M. fructicola , M. laxa and M. fructigena are the most common all over the world. Although they have been intensively studied, the availability of genomic and transcriptomic data in public databases is still scant. We sequenced, assembled and annotated the transcriptomes of the three pathogens using mRNA from germinating conidia and actively growing mycelia of two isolates of opposite mating types per each species for comparative transcriptome analyses. Results Illumina sequencing was used to generate about 70 million of paired-end reads per species, that were de novo assembled in 33,861 contigs for M. fructicola , 31,103 for M. laxa and 28,890 for M. fructigena . Approximately, 50% of the assembled contigs had significant hits when blasted against the NCBI non-redundant protein database and top-hits results were represented by Botrytis cinerea , Sclerotinia sclerotiorum and Sclerotinia borealis proteins. More than 90% of the obtained sequences were complete, the percentage of duplications was always less than 14% and fragmented and missing transcripts less than 5%. Orthologous transcripts were identified by tBLASTn analysis using the B. cinerea proteome as reference. Comparative transcriptome analyses revealed 65 transcripts over-expressed (FC ≥ 8 and FDR ≤ 0.05) or unique in M. fructicola , 30 in M. laxa and 31 in M. fructigena . Transcripts were involved in processes affecting fungal development, diversity and host-pathogen interactions, such as plant cell wall-degrading and detoxifying enzymes, zinc finger transcription factors, MFS transporters, cell surface proteins, key enzymes in biosynthesis and metabolism of antibiotics and toxins, and transposable elements. Conclusions This is the first large-scale reconstruction and annotation of the complete transcriptomes of M. fructicola , M. laxa and M. fructigena and the first comparative transcriptome analysis among the three pathogens revealing differentially expressed genes with potential important roles in metabolic and physiological processes related to fungal morphogenesis and development, diversity and pathogenesis which need further investigations. We believe that the data obtained represent a cornerstone for research aimed at improving knowledge on the population biology, physiology and plant-pathogen interactions of these important phytopathogenic fungi.

Several Aureobasidium spp. strains isolated from wild environments during winter 2022 were characterized by sequence analysis of the internal transcribed spacer (ITS), the translation elongation factor EF-1α gene (EF1), and part of the elongase gene (ELO). The variability in the EF1 and ELO loci are higher than in the ITS. All strains but one (UC14), were identified as A. pullulans. To assess the effectiveness of the characterized strains as biocontrol agents (BCAs) of diseases occurring during postharvest storage, a selection of the strains was evaluated by in vitro and in vivo assays. On average, the reduction of Monilinia spp. colony growth was more marked for non-volatile metabolites than for volatile (VOCs). Strain UC14 provided the strongest mycelial growth reduction of Monilinia fructicola by VOCs (66%). According to the in vivo results, all strains were effective in controlling brown rot during cold storage and remarkably in restricting the growth of Monilinia polystroma. In particular, VB23 was the most effective in controlling brown rot incidence, by 80%, 60%, 100%, and severity, by 79.5%, 72.7% and 100%, for Monilinia laxa, M. fructicola, and M. polystroma, respectively.

Monilinia fructicola, the most destructive pathogen of the genus Monilinia, has recently been introduced into Serbia and many other European countries. Since then, many studies have been conducted to evaluate the characteristics of Monilinia species that have a role in the establishment and survival of the pathogen in new areas. The present study assessed the capacity of M. fructicola to repress and replace Monilinia laxa in Serbia based on: fungicide sensitivity, growth rate and aggressiveness at different temperatures, as well as frost hardiness of the isolates of both species. The results showed that the isolates of M. fructicola, compared to M. laxa, were significantly less sensitive to the following fungicides: iprodione, tebucanozole, chlorothalonil, azoxystrobin, fluopyram, and boscalid. In addition, M. laxa isolates exhibited little variation in sensitivity to all of the tested fungicides, whereas M. fructicola isolates displayed a wide range of sensitivity. The temperature of 5°C favored M. laxa growth and aggressiveness, while at 30°C M. fructicola grew faster and had higher lesion expansion rate. These results support an assumption that M. fructicola will continue to spread in Serbian orchards in coming years, particularly on stone fruits harvested during hot summer weather.

Necrotrophic phytopathogenic fungi secrete degrading enzymes and phytotoxins to penetrate and colonise plant tissues. Since knowledge on the pathogenic processes of brown rot in fruit which becomes infected by Monilinia spp. is limited, we undertook an investigation whose aims were to detect: (i) the degrading enzymes of Monilinia spp. when cultured on media amended with different substrates, and (ii) the phytotoxins which are secreted by Monilinia fructicola, M. fructigena and M. laxa in brown rot-infected fruit. The sporulation and growth of 10 M. fructicola isolates, 9 M. fructigena isolates, and 10 M. laxa isolates plates, which contained 25 mL 2% (w/v) bacteriological agar, 0.67% (w/v) yeast nitrogen base, and 1% (w/v) of one of the following substrates: (1) β1–3 glucan (callose), (2) casein hydrolysate, (3) cutin, (4) methylcellulose, (5) apple pectin, (6) polygalacturonic acid sodium salt, (7) D(+)-sucrose, and (8) xylan were determined in order to detect the degrading enzymes. To look for the phytotoxins, the juice of nectarines that were inoculated with conidial suspensions M. fructicola, M. fructigena and M. laxa were subjected to liquid-liquid extraction with different organic solvents. The extracts were tested for their ability to cause necrosis on nectarine discs and the ones with a positive response were analysed by high performance liquid chromatography with DAD and MS spectrometer detectors and different fractions were collected to delimit metabolites. We found that Monilinia spp. secrete phytotoxins with a molar mass between 329 and 387 g mol−1 and degrading enzymes (cutinases, α-glucosidases, pectin lyases, polygalacturonases, proteases, and xylanases) for penetrating the fruit surface and invading and colonising fruit tissues.

Brown rot, caused by Monilinia spp., is a major peach disease worldwide. In this study, the response of peach cultivars Royal Glory (RG) and Rich Lady (RL) to infection by Monilinia fructicola or Monilinia laxa , was characterized. Phenotypic data, after artificial inoculations, revealed that ‘RL’ was relatively susceptible whereas ‘RG’ was moderately resistant to Monilinia spp. Comparative proteomic analysis identified mesocarp proteins of the 2 cultivars whose accumulation were altered by the 2 Monilinia species. Functional analysis indicated that pathogen-affected proteins in ‘RG’ were mainly involved in energy and metabolism, while, differentially accumulated proteins by the pathogen presence in ‘RL’ were involved in disease/defense and metabolism. A higher number of proteins was differentiated in ‘RG’ fruit compared to ‘RL’. Upon Monilinia spp. infection, various proteins were-down accumulated in ‘RL’ fruit. Protein identification by mass spectrometric analysis revealed that several defense-related proteins including thaumatin, formate dehydrogenase, S -formylglutathione hydrolase, CBS domain-containing protein, HSP70, and glutathione S -transferase were up-accumulated in ‘RG’ fruit following inoculation. The expression profile of selected defense-related genes, such as major latex allergen, 1-aminocyclopropane-1-carboxylate deaminase and UDP-glycoltransferase was assessed by RT-PCR. This is the first study deciphering differential regulations of peach fruit proteome upon Monilinia infection elucidating resistance responses.

Brown rot disease caused by fungal species from the Monilinia genus has recently been observed as one of the most important limiting factors for yield and quality of peach fruits in Turkey. During June and July of 2018, field trips to different peach orchards were performed in six provinces located in four different geographical regions of Turkey. One hundred and twenty-nine isolates were obtained from the diseased fruits, and of those, 109 were identified as Monilinia fructicola, whereas 20 were M. laxa according to the species-specific molecular markers. Mating types of the isolates were detected by PCR assays using primers designed in this study. Each isolate represented only one of the mating type genes. Overall data sets presented 1:1 ratio of mating types for both species, indicating a possible sexual reproduction. No polymorphisms were detected in partial sequences of mating type genes. The isolates were in vitro tested for their pathogenicity using apple fruits as a host and aggressiveness were evaluated at different time points using lesion sizes. Monilinia laxa was found more aggressive than M. fructicola. Moreover, M. fructicola isolates displayed larger colony growth compared to M. laxa isolates. However, no significant correlation was detected between the colony growth rates and virulence. This study is the first to demonstrate brown rot pathogens of peach fruit in Turkey and identifies different aspects of the pathogens, which would be useful in containment of pathogen spread and comparison with other populations of these pathogens worldwide, and suggests a new set of mating type markers for these species.

Stone fruit is attacked by various pathogens, of which brown rot disease is one of the important diseases. There are three Monilinia species mainly responsible for the brown rot disease: Monilinia fructicola is mainly found in North America and Australasia, and M. laxa and M. fructigena mainly in Europe. Both M. fructicola and M. laxa can infect flowers, resulting in blossom blight, as well as both healthy and wounded fruit, resulting in brown rot. On the other hand, M. fructigena can only infect wounded fruit. Compared to the two other species, M. fructicola has been extensively studied, whereas the equally important M. laxa has had less attention. This paper addresses this imbalance and reviews research on the biology, epidemiology and management of M. laxa on stone fruits. Due to EU regulations, the number of fungicides available for controlling plant diseases has been steadily decreasing, particularly in the post-harvest environment. This has placed much more emphasis on alternative control methods, a focus of the present review. Numerous physical and biological approaches to control have achieved successful outcomes but often in small-scale trials and in isolation from integrated strategies. Promising physical control methods include removal of mummified fruit in orchards and post-harvest hot-water treatment. Many micro-organisms have been shown to have biocontrol potential against brown rot but only a few have been commercially formulated. It is generally agreed that the use of biocontrol agents needs to be integrated with other measures. Current research focuses on disease management from flowering to post-harvest period. Recent results have suggested that reducing overwintering inoculum should be considered as one of key aspects of integrated management of brown rot on stone fruit. Finally, we make recommendations about future research and development on integrated pest management strategies for control of M. laxa, especially on strategic deployment of biocontrol agents and interactions among brown rot pathogens.

The reciprocal targeting of microRNAs (miRNA) and micro-like-RNAs (milRNA) between hosts and pathogens is critical for understanding their interactions. In this study, reciprocal miRNA targets were explored in two Monilinia fungal pathogens, M. fructicola & M. laxa, and their peach host (Prunus presica). Using in silico analysis, 355 and 266 putative miRNAs were predicted for M. fructicola and M. laxa, respectively. Number of miRNAs and their targets differed based on host and pathogen species as 209 M. fructicola miRNAs target 98 peach genes and 128 M. laxa miRNAs target 338 peach genes. On the other hand, peach miRNAs showed the species-specific responses targeting fungal pathways to struggle with its pathogens. These findings indicate distinct strategies and species-specific interactions in this pathosystem. Besides, through the in vitro experimental designs, 166 and 124 expressed miRNAs by M. fructicola were detected in the host-mimicked and control environments, respectively. Additionally, novel miRNAs were discovered, six of which were in the mimicked environment and the seven in the controlled environment as highlighting dynamic and specialized miRNA expression in M. fructicola depending on the environmental conditions. In conclusion, this study provides the first insights into miRNA-mediated interactions between M. fructicola, M. laxa, and peach hosts. Unrevealing the cross talk through the miRNAs in host–pathogen interactions enhances the understanding of pathogenesis and host defense mechanisms. These findings have implications for disease management strategies and contribute to the fields of basic science and evolutionary biology.

Benzimidazoles, dicarboximides and demethylation inhibitors are the main group of fungicides used to control brown rot in Spain. The causal agents of brown rot in peach (Prunus persica (L.) Batsch) orchards in Spain are Monilinia laxa M. fructigena, and M. fructicola. The dynamics of fungicide sensitivity and fitness of M. fructicola population, the most recent species causing brown rot in the Ebro Valley (Lleida, Spain), were characterized by determining their resistance, fitness, and in vitro competitiveness to thiophanate-methyl (TM), iprodione (I), and cyproconazole (CPZ) in field isolates recovered over the 2006–2010 period. We found that: (a) more than 95 % of the M. fructicola isolates are high TM-resistant, (b) more than 50 % of the M. fructicola isolates are I-resistant and these frequencies of occurrence did not change during our 5-year survey, and (c) a few CPZ-resistant isolates have been also detected in population since 2008. We identified five different fungicide-resistant (R) and/or fungicide-sensitive (S) phenotypes and found that our study population contains multiple fungicide-resistant isolates. Moreover, these fungicide-resistant isolates display high parasitic fitness on fruit and flowers and high competitiveness. These findings suggest that the TM, I, and CPZ resistance of M. fructicola isolates could be contributing as another factor on changing the frequency of occurrence of the three Monilinia species in the Ebro Valley.