Pathogenic Fungal Species Research Articles

Long amplicon nanopore sequencing of Botrytis cinerea and other fungal species present in infected grapevine leaf samples.

Botrytis cinerea is a well-known plant pathogen responsible for grey mould disease infecting more than 500 plant species. It is listed as the second most important plant pathogen scientifically and economically. Its impact is particularly severe in grapes since it affects both the yield of grape berries and the quality of wines. While various methods for detecting B. cinerea have been investigated, the application of Oxford Nanopore Technology (ONT) for complete ribosomal operon sequencing, which has proven effective in human and animal fungal research and diagnostics, has not yet been explored in grapevine (Vitis vinifera) disease research. In this study, we sequenced complete ribosomal operons (∼5.5 kb amplicons), which encompass the 18S, ITS1, 5.8S, ITS2, and 28S regions, from both pure cultures of B. cinerea and infected grapevine leaf samples. Minimap2, a sequence alignment tool integrated into the EPI2ME software, served as a taxonomy classifier, utilizing the custom reference database FRODO. The results demonstrate that B. cinerea was detectable when this pathogen was not the dominant fungal species in leaf samples. Additionally, the method facilitates host DNA-free sequencing and might have a good potential to distinguish other pathogenic and non-pathogenic fungal species hosted within grapevine's infected leaves, such as Alternaria alternata, Saccharomyces cerevisiae, Saccharomyces boulardii, Mucor racemosus, and Ascochyta rabie. The sequences were uploaded to the NCBI database. Long amplicon sequencing method has the capacity to be broadened to other susceptible crops and pathogens, as a valuable tool for early grey rot detection and mycobiome research. Future large-scale studies are needed to overcome challenges, such as comprehensive reference databases for complete fungal ribosomal operons for grape mycobiome studies.

High-quality genome resource of Lasiodiplodia pseudotheobromae associated with die-back on Eucalyptus trees.

Lasiodiplodia pseudotheobromae is an important fungal pathogen associated with die-back, canker and shoot blight in many plant hosts with a wide geographic distribution. The aim of our study was to provide high-quality genome assemblies and sequence annotation resources of L. pseudotheobromae, to facilitate future studies on the systematics, population genetics and genomics of the fungal pathogen L. pseudotheobromae. High-quality genomes of five L. pseudotheobromae isolates were sequenced based on Oxford Nanopore technology (ONT) and Illumina HiSeq sequencing platform. The total size of each assembly ranged from 43Mb to 43.86Mb and over 11,000 protein-coding genes were predicted from each genome. The proteins of predicted genes were annotated using multiple public databases, among the annotated protein-coding genes, more than 4,300 genes were predicted as potential virulence genes by the Pathogen Host Interactions (PHI) database. Moreover, the genome comparative analysis among L. pseudotheobromae and other closely related species revealed that 7,408 gene clusters were shared among them and 152 gene clusters unique to L. pseudotheobromae. This genome and associated datasets provided here will serve as a useful resource for further analyses of this fungal pathogen species.

Evaluation of pests affecting maize imported across Malawi borders

Cross-border trade is one of the major factors that puts maize at risk as it creates pathways for spread of different maize pests in Malawi. This study was carried out to identify pests associated with imported maize in order to improve phytosanitary measures and reduce the introduction of pests. Random sampling technique was used to collect imported maize at selected borders and commercial cities with the aim of identifying pests present during importation. There were no significant differences (p>0.05) in the population of insect pests before and after incubation in all the surveyed districts. Significant differences were observed after incubation for larger grain borer (LGB) and common maize weevil. The highest percentage infestation (32%) of living LGB was recorded in Karonga and the least in Mzuzu (p<0.05). On the other hand, for maize weevil, the highest percentage infestation (20%) was recorded in Nkhatabay while the least was observed in Lilongwe district (p<0.05). Common fungal pathogen species isolated included Fusarium (70%), Aspergillus (29%) and Penicillium (1%). The highest percentage of kernel infection with Fusarium sp. was recorded in Dedza and Lilongwe while for Aspergillus sp. were in Mzuzu. The Aspergillus sp. isolated were A. niger (29%), A. flavus (22%) and A. parasiticus (5%) while Fusarium sp. isolated was Fusarium verticilloides. The study provided the status and causes of storage losses by various pests on maize. It is recommended that the phytosanitary system present at various Malawi borders where maize is imported should be strengthened. This can be achieved through human and infrastructure capacity building, strict compliance with importation regulations and improved funding of organizations mandated to ensure phytosanitary compliance. Key words: Aspergillus sp., consignment, food security, grain storage, random sampling, sanitary and phytosanitary measures.

Molecular phylogeny and morphology reveal two new graminicolous species, Curvularia aurantiasp. nov. and C. vidyodayana sp. nov. with new records of Curvularia spp. from Sri Lanka.

Despite being a small island, Sri Lanka is rich in fungal diversity. Most of the fungi from Sri Lanka have been identified as pathogens of vegetables, fruits, and plantation crops to date. The pleosporalean genus Curvularia (Dothideomycetes) includes phytopathogenic, saprobic, endophytic, and human/animal opportunistic pathogenic fungal species. The majority of the plant-associated Curvularia species are known from poaceous hosts. During the current study, 22 geographical locations of the country were explored and collections were made from 10 different poaceous hosts. Morphology and molecular phylogeny based on three loci, including nuclear internal transcribed spacers 1 and 2 with 5.8S nrDNA (ITS), glyceraldehyde-3-phosphate dehydrogenase (gapdh), and translation elongation factor 1-α (tef1) supported the description of two new species of fungi described herein as C. aurantia sp. nov. and C. vidyodayana sp. nov. Moreover, novel host-fungal association records for C. chiangmaiensis, C. falsilunata, C. lonarensis, C. plantarum, and C. pseudobrachyspora are updated herein. In addition, five species within the genus Curvularia, viz., C. asiatica, C. geniculata, C. lunata, C. muehlenbeckiae, and C. verruculosa represent new records of fungi from Sri Lanka. Citation: Ferdinandez HS, Manamgoda DS, Udayanga D, Munasinghe MS, Castlebury LA (2023). Molecular phylogeny and morphology reveal two new graminicolous species, Curvularia aurantia sp. nov. and C. vidyodayana sp. nov. with new records of Curvularia spp. from Sri Lanka. Fungal Systematics and Evolution 12: 219-246. doi: 10.3114/fuse.2023.12.11.

Synthesis, antimicrobial, anticancer evaluation and molecular docking with Bax and MDM2 of dibromosterculic acid

Dibromosterculic acid [8-(1,2-dibromo-2-octylcyclopropyl)-octanoic acid], a new synthetic derivative was prepared by bromination of sterculic acid. This synthetic derivative showed strong fungicidal activity against two pathogenic fungal species namely Penicillium chrysogenum and Aspergillus niger with minimum inhibitory concentration (MIC) value of 0.007 mg/ml and good bactericidal activity against Bacillus subtilis and Xanthomonas sp. with MIC value of 0.015 mg/ml. Cytotoxic activity on both normal (MCF-10A) and cancerous (MDA-MB-468) cell lines revealed that the survivability percentage of normal cells was unaffected, whereas cancerous cells were decreased greatly by dibromosterculic acid with 50% survivability at 9 µg/ml concentration. Molecular-docking using AutoDock 4.2 with Bax exhibited strong pi-sigma interaction with PHE-93, pi-alkyl and alkyl interaction with TRP-139, ARG-89 and PHE-92 whereas MDM2 revealed strong hydrogen bond interaction with GLN-59 and pi-alkyl interaction with PHE-55. All experimental parameters suggested that this synthetic derivative would be valuable for target-specific drug development with nominal side effects.

Genetic diversity and pathogenicity of fungi associated with rice sheath rot in the Fogera plain, Northwest Ethiopia

AbstractSheath rot is a widespread rice disease that affects rice crops worldwide. It is particularly prevalent in Ethiopia, with the Fogera plains exhibiting the highest incidence and severity. However, the lack of reliable molecular‐level identification of rice fungal pathogens in Ethiopia has hindered the development of effective management practices. This study aimed to assess the diversity of fungal agents causing sheath rot disease in rice crops around the Fogera plains, Ethiopia, using both morphology and nucleotide sequences of the ITS1 rDNA region. Additionally, the pathogenicity of the isolates was evaluated using attached tiller assay methods. A total of 16 fungal isolates were recovered from diseased plant samples. Among these, five species; Phoma spp., Phoma herbarium, Epicoccum sorghinum, Epicoccum lantuscollum and Microdochium seminicola were identified and reported as new records of sheath rot disease in rice. The predominant genus was Phoma (75%) followed by Epicoccum (18.75%). The pathogenicity caused by E. sorghinum was particularly severe and significantly different from the symptoms caused by other isolates, resulting in a mean lesion length of 167.7 mm and a disease severity of 55.7%. These findings suggest that E. sorghinum is the most important rice pathogen in the region. Our results have identified the most predominant and pathogenic fungal species associated with sheath rot disease in the region. Importantly, this study marks the first report of sheath rot‐causing fungi identified in rice using the ITS rDNA‐sequencing technique in Ethiopia. Therefore, this research may contribute to the development of fungal disease management strategies in rice and provide evidence to initiate a rice breeding program focused on developing sheath‐rot‐resistant rice varieties.

РОЛЬ МСКТ И МРТ В ДИФФЕРЕНЦИАЛЬНОЙ ДИАГНОСТИКЕ ОСЛОЖНЕНИЙ СРЕДНЕЙ ЗОНЫ ЛИЦА У ПАЦИЕНТОВ С ПАТОЛОГИЕЙ ГРИБКОВОЙ ЭТИОЛОГИИ

Abstract. Introduction. Fungi are one of the constituents of an ecosystem. There are about 250 known human- pathogenic fungi species: From the widely spread Candida to the pathogene of aspergillosis that may lead to fatal consequences in certain conditions. Studies performed by Russian and foreign authors show the increase in the frequency of chronic sinusitis determined by the presence of fungi or fungal-microbial associations. Aim of this study is to perform the differential diagnosis of midface complications in patients with fungal pathology according to multispiral computed tomogrsphy and magnetic resonance tomography. Materials and Methods. A retrospective study was conducted based on the findings of multispiral computed and/or magnetic resonance tomographies of 68 patients. Presence of a fungal process was confirmed by histopathologic, mucroscopic, and cultural investigations. Results and Discussion. Histological investigations of surgical specimen detected the presence of fungal bodies formed by mycelium, i.e., a cluster of densely, compactly located fungal flocci in 28 of 48 (58.3 %): In 28 patients with maxilla sinus lesions, in 14 patients with sohenoidal sinus lesions, and in 5 patients with ethmoidal sinus lesions. In case of acute invasive fungal rhinosinusitis, the computed tomography of paranasal sinuses detected mucosal thickening and the bone density decrease associated with an erosive process. Unilateral lesions, erosions, and thinning of the sinus bone walls. According to magnetic resonance tomography, there was low signal in T2 mode, which is determined by the high concentrations of various metal elements in fungi, as well as by a high protein level and low free water contents in mucine. Conclusions. Visualization is essential to staging the disease and assessing the involvement of deeper structures, which may be clinically unmanifest. Although both computed tomography and magnetic resonance tomography are useful, the contrast-enhanced magnetic resonance tomography is the method of choice and helps assess the spread of the disease, especially regarding the deep extension into maxilla-facial area and brain.

Seasonal Variation in Fungi in Beach Sand in Summertime: Stintino (Italy)

Background: The goal of this study was to monitor the microbial biodiversity in beach sand that is heavily visited by tourists during the summer, and to determinate whether the high presence of bathers (around 5000 per day) can modify sand microbial composition. Methods: Between 2016 and 2020, 150 sand samples were collected from nine different points at La Pelosa beach in Sardinia, Italy. Non-culturing methods were used; DNA extraction and meta-barcode sequencing were performed. All samples were analyzed with sequencing methods for 16S and ITS sequences. Results: Fungal genera differ on the three beaches and in the winter/summer zones. The ITS sequence showed the most common presence of Candida during summer and Paradendryphiella in the winter. The greatest diversity was found in the dune during winter, while in other parts of the beach, there are differences between bacteria and fungi, particularly in the wash zone during the winter, with high diversity for 16S sequences but low diversity for ITS sequences. Conclusions: It appears reasonable that the sands, even on non-urban beaches, should be included in health monitoring programs in addition to the waters, and that access to them should be regulated by limiting the number of bathers with the aim of reducing the presence of pathogenic fungal species.

Determination of Antifungal Activity of Leaf Extracts from Hypericum ssp. Against Plant Pathogenic Fungi Fusarium oxysporum and Alternaria Alternata

In this study, the antifungal activity of ethanol extracts of three plant species of Hypericum perforatum, Hypericum scabrum and Hypericum origanifolium were evaluated for against two plant pathogenic fungal species of Fusarium oxysporum and Alternaria alternata.The antifungal activities of Hypericum ssp. extracts (3, 15, 45, 80, 120, 200, 240, 300 ppm) were tested against fungi. Ethanol 30% as control; Commercial fungicide (1 μg/mL) was used as a positive control. All Hypericum ssp. plant extracts were effective in different rates against the fungi depending on the doses. In addition, IC50 values of the three selected plant Hypericum ssp. leaf extract against pathogenic fungus F.oxysporum and A. alternata were determined with of probit analysis. In terms of the IC50 values, the antifungal activity of H. perforatum plant exract against F.oxysporum (122.525 μg/mL) > A.alternata (108.484 μg/mL). On the other hand, IC50 values of H. scabrum against A.alternata (126.390 μg/mL) > F.oxysporum (113.714 μg/mL). Moreover, the IC50 value of H. origanifolium against two pathogenic fungi was calculated as F.oxysporum (159.931 μg/mL) > A.alternata (55.759 μg/mL). Results showed that H.origanifolium has the best fungicidal activity, with IC50 value of 55.759 μg/mL against A.alternata and H.scabrum 113.717 μg/mL against F.oxysporum. However, the highest concentrations (300 ppm) caused completely inhibition in the both fungi mycelial growth followed by lower concentrations of plant extracts. A. alternata were the sensitive fungal species, while the F. oxysporum were a more resistant to the Hypericum ssp. extracts. Based on the antifungal activity tests, these plants extracts this may have effective as the new natural fungicide protecting crops against fungal diseases.

Molecular and machine learning approaches to study the impact of climatic factors on the evolution of cranberry fruit rot

IntroductionCranberry (Vaccinium macrocarpon) is an important crop grown in the United States and Canada, with the province of Québec being the world’s largest producer of organic cranberry. However, cranberry fruit rot (CFR), caused by 12 fungal species, has become a major issue affecting yield.MethodsA molecular detection tool was used to detect the presence of the 12 CFR fungi and evaluate CFR species across three farms with different fungicide strategies in Québec. The incidence and frequency of CFR fungi were evaluated for 2020 and compared with 2018 data from the same farms in Québec. Machine-learning models were used to determine the possibility of explaining CFR disease and species based on climate, and analyze the effects of weather variables on CFR presence andprimary fungal species.ResultsThe most predominant CFR species remained the same in both years, with Godronia cassandrae and Coleophoma empetri being the two most common, but some species showed changes in relative abundance. Furthermore, this study examined the diversity variations in 2018 and 2020, with data showing an overall increase in diversity over the period. The results showed that fungicide applications impacted the species composition of CFR among the farms. Five weather variables (daily snow on the ground (cm), total daily precipitation (mm), daily atmospheric pressure (kPa), daily relative humidity (%) and daily temperature (°C)) were selected and found to contribute differently to the model with atmospheric pressure being the most important. Surprisingly, temperature and precipitations did not influence much the incidence of fungal pathogen species and each CFR species behaved differently in response to environmental factors.DiscussionOverall, this study highlights the complexity of predicting CFR disease, as caused by 12 fungi, and of developing effective disease management strategies for CFR.

Research progress and management strategies of fungal diseases in Camellia oleifera

Camellia oleifera Abel, a woody oil plant, that is endemic to China. Tea oil, also referred to as “oriental olive oil,” is a superior quality plant-based cooking oil. The production of tea oil accounts for 8% of the total edible vegetable oil production in the country. Since 2022, the annual output value of C. oleifera industry has exceeded 100 billion yuan, making it one of the major economic contributors to China’s rural revitalization development strategy. In recent years, demand and production have grown in parallel. However, this has led to an increase in the incidence levels of pest and diseases. Pests and diseases significantly reduce the quality and yield of C. oleifera. C. oleifera diseases are mainly caused by pathogenic fungi. C. oleifera anthracnose, soft rot, leaf spot, coal stain, leaf gall disease, and root rot are the most important fungal diseases affecting the C. oleifera industry. However, the same disease may be caused by different pathogenic fungi. C. oleifera can be found in half of China and is found in several climatic zones. The geographical distribution of woody plant diseases is consistent with the distribution of the tree species and the ecology of the range, which also results in a highly complex distribution of fungal diseases of C. oleifera. The management of fungal diseases in C. oleifera is extremely challenging due to the variety of pathogenic fungal species, multiple routes of transmission, the lack of resistant plants, and the environmental safety of chemical measures. The optimal strategy for addressing fungal diseases in C. oleifera is to develop and apply an integrated disease management plan. This review provides a brief overview of the pathogenic species, pathogenesis, pathogenesis, geographical distribution, current management strategies, and potentially new methods of C. oleifera fungal diseases, to provide direction for the development of comprehensive management measures for C. oleifera fungal diseases in the future.

Effects of Pre-Harvest Spraying with Salicylic Acid (SA) and Sodium Nitroprusside (SNP) on Storage Quality and Pathogenic Fungal Species in ‘Manaohong’ Cherries

Chinese cherries, with their delightful blend of sourness and sweetness, are highly favored for their taste and nutritional benefits. However, they mature in conditions of high temperatures and rainfall, making them vulnerable to fungal infections which compromise their post-harvest quality. Our research aimed to study the effects of pre-harvest spraying with salicylic acid (SA) and sodium nitroprusside (SNP) on the pathogenic fungi in Manaohong cherries and their subsequent storage quality. We discovered that using SA and SNP at varying fruit development stages preserved fruit hardness, texture, appearance, and respiratory rate during storage, although it did not significantly alter the fruit’s dimensions. Furthermore, this pre-harvest treatment preserved levels of titratable acids, total phenols, and other antioxidants in the cherries, bolstered the activities of certain antioxidant enzymes (SOD, APX), and inhibited the activity of PPO and POD enzymes.Notably, the SA treatment alone demonstrated superior storage performance compared to combined treatments. Our research also identified Alternaria alternata and Colletotrichum godetiae as the primary pathogens in Manaohong cherries. In in vitro experiments, neither SA nor SNP inhibited these fungi’s growth. Consequently, we evaluated 12 pesticides and determined that 5% hexaconazole and 50% benomyl were most effective against these pathogens. Thus, to enhance the shelf life of Manaohong cherries and ensure their post-harvest quality, we recommend a pre-harvest spray of a SA, and combined with 5% hexaconazole and 50% benomyl. This approach not only promises enhanced cherry longevity but also lays a foundational strategy for the flourishing Manaohong Cherry industry.

Design, Synthesis and Antifungal Evaluation of N-Substituted Maleimide Derivatives with Imidazole and 2-Methyl Imidazole Moieties

A series of N-substituted maleimide derivatives with an attached imidazole and 2-methyl imidazole heterocyclic rings were designed, synthesized and evaluated for their antifungal activity against four pathogenic fungi. 1H-NMR, 13C-NMR and mass spectra confirmed the chemical structures of all synthesized compounds. All compounds 4a-4g, 5b and 5f were initially screened for qualitative (zone of inhibition) antifungal activity against C. albicans, A. fumigatus, A. niger, and A. oryzae. The screening results indicate that most of the synthesized compounds showed significant antifungal activity against the tested fungi. Furthermore, the compounds that showed a significant zone of inhibition were selected and tested quantitatively (MIC50 and IC50) against the same pathogenic fungi species. The MIC50 and IC50 results of selected compounds were analyzed. These results strongly suggest that compound 5f has shown promising antifungal activity. Furthermore, the structure–activity relationship of compound 5f revealed that the compound substituted with the –F group possess prominent antifungal activity.

Recognising ceratocystis disease symptoms in a Eucalyptus pellita nursery

Early recognition of symptoms of infection is essential to prevent diseases from spreading through nurseries and into planted systems. In plantations, symptoms of Ceratocystis manginecans infection are typically wilt and canker but symptoms may differ at the nursery stage and as yet have not been described for Eucalyptus pellita. In this study, disease symptoms were recorded over a three-month period in an E. pellita nursery and isolates trapped. Then, to clearly identify those relating to ceratocystis disease and any related mortality, two C. manginecans isolates were re-inoculated by either spraying or dipping of healthy plantlets raised by tissue culture and mini-cuttings. Many disease symptoms were observed in the nursery but only perithecia on the leaves and stems was a clear indication of the presence of a Ceratocystis pathogen. Nine pathogenic fungal species, including C. manginecans, were isolated and identified by DNA analysis. Koch’s postulates indicated that leaf blight associated with C. manginecans was black-brown in colour, irregular, with uneven edges and surrounded by a yellow margin or halo on the upper leaf surface. Mortality following inoculation of plantlets by spraying immediately after planting was > 70%; by contrast, mortality of mini-cuttings was < 50% if spraying was done just before transplanting but < 20% if done 7 or 14 days after transplanting. Isolate EP313C caused significantly greater mortality than isolate EP106C. The research showed that C. manginecans and other pathogens were present in the nursery environment and must be identified and managed as part of good nursery practice.

Co-inoculation timing affects the interspecific interactions between phoma stem canker pathogens Leptosphaeria maculans and Leptosphaeria biglobosa.

Phoma stem canker is an economically important disease of oilseed rape, caused by two co-existing fungal pathogen species, Leptosphaeria maculans (Plenodomus lingam) and Leptosphaeria biglobosa (Plenodomus biglobosus). Leptosphaeria maculans produces a phytotoxin called sirodesmin PL. Our previous work showed that L. biglobosa has an antagonistic effect on the production of sirodesmin PL if it is simultaneously co-inoculated with L. maculans. However, the effects of sequential co-inoculation on interspecific interactions between the two pathogens are not understood. The interactions between L. maculans and L. biglobosa were investigated in liquid culture by inoculation with L. maculans first, followed by L. biglobosa sequentially at 1, 3, 5 or 7 days later and vice versa; the controls were inoculated with L. maculans only, L. biglobosa only, or L. maculans and L. biglobosa simultaneously. The results showed that L. biglobosa inhibited the growth of L. maculans, the production of both sirodesmin PL and its precursors if L. biglobosa was inoculated before, or simultaneously with, L. maculans. However, the antagonistic effects of L. biglobosa were lost if it was co-inoculated 5 or 7 days after L. maculans. For the first time, the results of this study provided evidence that the timing when L. maculans and L. biglobosa meet significantly influences the outcome of the interspecific competition between them. Leptosphaeria biglobosa can inhibit the production of sirodesmin PL and the growth of L. maculans if it is inoculated before L. maculans or less than 3 days after L. maculans in liquid culture. There is a need to further investigate the timing of co-inoculation on interactions between L. maculans and L. biglobosa in their host plants for improving the control of phoma stem canker. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Pathogenicity of entomopathogenic fungi to eggs, larvae, and adults and their effects on development of Aedes albopictus

Abstract. Ramayanti I, Herlinda S, Muslim A, Hasyim H, Anwar C, Suwandi S, Damiri N, Irsan C, Verawaty M. 2023. Pathogenicity of entomopathogenic fungi to eggs, larvae, and adults and their effects on development of Aedes albopictus. Biodiversitas 24: 4766-4774. No information is available on the effect of fungi on the development of Aedes albopictus and the effectiveness of fungi in killing its eggs, larvae, and adults. The aim of this research was to determine the pathogenicity of entomopathogenic fungi on the eggs, larvae, and adults of Ae. albopictus and to investigate the effects of the fungi on the development of Ae. albopictus. The fungal species identified molecularly were Aedes albopictus, Beauveria bassiana, Metarhizium anisopliae, Penicillium citrinum, and Talaromyces diversus. The results showed that entomopathogenic fungi negatively affected the development of Ae. albopictus and could decrease the eggs laid by adult females and shorten adult longevity. M. anisopliae was the most pathogenic fungal species (mortality rate 100%) for eggs, larvae, pupae, and adults of Ae. albopictus. However, B. bassiana, P. citrinum, and T. diversus were also found pathogenic to them. The effect of eggs treated with entomopathogenic fungi negatively affected the larval, pupal, and adult stages of Ae. albopictus. These findings highlighted that M. anisopliae, B. bassiana, P. citrinum, and T. diversus have insecticidal activity against Ae. albopictus eggs, larvae, pupae, and adults. Therefore, further investigation is needed to develop these fungal species into ovicides, larvicides, and adulticides to control Ae. albopictus.

The role of the Mediator complex in fungal pathogenesis and response to antifungal agents.

Mediator is a complex of polypeptides that plays a central role in the recruitment of RNA polymerase II to promoters and subsequent transcriptional activation in eukaryotic organisms. Studies have now shown that Mediator has a role in regulating expression of genes implicated in virulence and antifungal drug resistance in pathogenic fungi. The roles of specific Mediator subunits have been investigated in several species of pathogenic fungi, particularly in the most pathogenic yeast Candida albicans. Uniquely, pathogenic yeast also present several interesting examples of divergence in Mediator structure and function, most notably in C. glabrata, which possesses two orthologues of Med15, and in C. albicans, which has a massively expanded family of Med2 orthologues known as the TLO gene family. This review highlights specific examples of recent progress in characterizing the role of Mediator in pathogenic fungi.

Rapid, efficient auxin-inducible protein degradation in Candida pathogens.

A variety of inducible protein degradation (IPD) systems have been developed as powerful tools for protein functional characterization. IPD systems provide a convenient mechanism for rapid inactivation of almost any target protein of interest. Auxin-inducible degradation (AID) is one of the most common IPD systems and has been established in diverse eukaryotic research model organisms. Thus far, IPD tools have not been developed for use in pathogenic fungal species. Here, we demonstrate that the original AID and the second generation, AID2, systems work efficiently and rapidly in the human pathogenic yeasts, Candida albicans and Candida glabrata. We developed a collection of plasmids that support AID system use in laboratory strains of these pathogens. These systems can induce >95% degradation of target proteins within minutes. In the case of AID2, maximal degradation was achieved at low nanomolar concentrations of the synthetic auxin analog 5-adamantyl-indole-3-acetic acid. Auxin-induced target degradation successfully phenocopied gene deletions in both species. The system should be readily adaptable to other fungal species and to clinical pathogen strains. Our results define the AID system as a powerful and convenient functional genomics tool for protein characterization in fungal pathogens. IMPORTANCE Life-threatening fungal infections are an escalating human health problem, complicated by limited treatment options and the evolution of drug resistant pathogen strains. Identification of new targets for therapeutics to combat invasive fungal infections, including those caused by Candida species, is an urgent need. In this report, we establish and validate an inducible protein degradation methodology in Candida albicans and Candida glabrata that provides a new tool for protein functional characterization in these, and likely other, fungal pathogen species. We expect this tool will ultimately be useful for the identification and characterization of promising drug targets and factors involved in virulence and drug resistance.

Nano-emulsification essential oil of Monarda didyma L. to improve its preservation effect on postharvest blueberry.

The reduction in blueberry harvest due to pathogen infection was reported to reach 80%. Essential oil (EO) can provide a new way to preserve blueberry. Here, in search for plants volatiles with preservation ability, a novel device was designed for the screening of aromatic plants led to the discovery of hit plant Monarda didyma L. Consequently, antifungi activity of M. didyma EO (MEO) and its nano-emulsion (MNE) were tested. 2 species of pathogenic fungi were isolated from blueberries, namely Alternaria sp. and Colletotrichum sp. were used as the target strains. In the in vitro activity test, the pathogenic were completely inhibited when the EO was 4µL or 1.0µL/mL. Compared with EO, MNE exhibited superior antimicrobial activity. Moreover, MNE can cause serious morphological changes and result in a decrease in the rot and weightlessness rate of blueberry. Hence, NME represents a promising agent for the preservation of postharvest blueberry.

Fungi complicit in oak powdery mildew infection in the Oława Forest District

Oaks are important forest-forming species in Poland. It is an important species from an economic perspective in Polish forestry. Oak is characterized by high tolerance in terms of soil requirements, which include soil formations from sandy to clayey with a reaction from acidic to indifferent, sometimes even alkaline in pedunculate oak. It is sensitive to late frosts. In recent years, climate change has been observed in Europe and also in Poland. Abiotic stresses caused by unfavourable weather conditions have worsened the health of oak trees, on which other pathogens, primarily fungi, have become established. The research was carried out at two sites in the Oława Forest District in Bystrzyca in 2022. They consisted in collecting 10 representative shoots from each research area, which were then used to isolate fungal species in the laboratory. The results of the experiment confirm that co-occurring species can be observed with the presence of powdery mildew. Oak shoots are colonised by both pathogenic and saprotrophic fungal species. The high occurrence of pathogenic fungi species along with powdery mildew on oak shoots can be associated with the deterioration of the phytosarcoma condition of oaks, however, in order to be able to state this unequivocally, research on this phenomenon should be focused on in the future.