Fungal Propagules Research Articles

Speleomycology of Air in Stopića Cave (Serbia).

Fungi can colonize organic matter present in subterranean sites and have a significant role as dwellers in different microniches of cave habitats. In order to analyze the content of airborne fungal propagules in different parts of "Stopića Cave," a touristic site in Serbia, air sampling was carried out in three seasons during 2020, prior to and during the onset of COVID-19 pandemic. Culturable mycobiota was identified using both microscopic techniques and ITS region/BenA gene barcoding, while multivariate analyses were employed to establish the link between fungal taxa and different environmental factors. The maximal measured fungal propagule concentrations were recorded during spring sampling which were based on fungal propagule concentration categories; the cave environment matches the category V. A total of 29 fungal isolates were identified, while Aspergillus, Cladosporium, Fusarium, Lecanicillium, Mucor, and Penicillium were the most diverse genera. According to the trophic mode, most of the isolated fungal species were pathotrophs (75.86%), but when regarding ecological guilds, the most dominant were undefined saprobes and animal pathogens (41.38% for each). Show caves are especially vulnerable to human impacts, and the fungal propagules' concentration within the caves could be good indices for the level of ecological disturbance.

Characterization of Aspartic Proteases from Paracoccidioides brasiliensis and Their Role in Fungal Thermo-Dimorphism

Paracoccidioidomycosis (PCM) is the most prevalent systemic mycosis in Latin America and is caused by fungi from the Paracoccidioides genus. The infection begins after inhalation of the fungal propagules and their thermo-dimorphic shift to yeast form. Proteases play an important role in the host invasion process and immune modulation in many pathogenic microorganisms. Aspartyl proteases are virulence factors in many human fungal pathogens that play an important role in the host invasion process morphogenesis, cellular function, immunity, and nutrition. In the present study, we characterized the modulation of acid proteases from Paracoccidioides brasiliensis. We detected four aspartyl proteases in P. brasiliensis with high homology to aspartic protease from Saccharomyces cerevisiae Pep4. Furthermore, we demonstrated that Pepstatin A can inhibit dimorphic switching (mycelium→yeast) in P. brasiliensis. In addition, these genes were modulated during thermo-dimorphism (M→Y transition) in the presence or absence of carbon and nitrogen sources and during growth at pH 4 during 24 and 48 h. We also observed that P. brasiliensis increase the secretion of aspartic proteases when cultivated at pH 4, and these acid proteases cleave BSA, collagen, and hemoglobin. These data suggest that aspartyl proteases are modulated by environmental conditions and during fungal thermo-dimorphism. Thus, this work brings new possibilities for studying the role of aspartyl proteases in the host-pathogen relationship and P. brasiliensis biology.

Development of New Preventive Strategies for Pine Pitch Canker Caused by Fusarium circinatum in Irrigation Water and Evaluation in a Real Nursery Context

Fusarium circinatum is one of the many threats to forests and Pinus nurseries all over the world, being classified as a quarantine organism by several organizations and governing bodies, such as the European and Mediterranean Plant Protection Organization (EPPO) and the European Union (EU), with associated phytosanitary measures in place to prevent its spread through the various means of dispersal. One such means of dispersal is the water used for irrigation in nurseries, which can contain fungal propagules. Three different treatments, namely, Desogerme, Intra Hydrocare and sodium hypochlorite (NaClO), were tested for their efficacy in eliminating F. circinatum spores in water at several concentrations. Those that showed 100% disinfection rates were selected for further assays regarding seed germination and water quality impact. From these studies, Desogerme 1% and Intra Hydrocare 4% were then selected for large-scale seed germination and plant certification assays in nurseries, where they showed promising results in regard to the prevention of infections in nurseries, and in this way, contribute to the efforts of mitigating this disease.

Production of Escovopsis weberi (Ascomycota: Hypocreales) Mycelial Pellets and Their Effects on Leaf-Cutting Ant Fungal Gardens

The maintenance of the symbiosis between leaf-cutting ants and their mutualistic fungus Leucoagaricus gongylophorus Singer (Moller) is vital for the survival of both species. The specialist fungal parasite Escovopsis weberi Muchovej & Della Lucia is a threat to this symbiosis, causing severe damage to the fungal garden. Mycelial pellets are resistant fungal structures that can be produced under laboratory conditions. These structures were studied for use in biological pest control, but the production of mycelial pellets has not previously been documented in Escovopsis. One of the aims of this study was to induce Escovopsis weberi to produce mycelial pellets and investigate the potential of these pellets for the control of leaf-cutting ants. We compared the pathogenicity of Escovopsis weberi mycelial pellets and conidia against mini-colonies of Acromyrmex subterraneus subterraneus Forel when applied in the form of baits. Worker ants were able to distinguish mycelial pellets from conidia, as baits with mycelial pellets were more attractive to workers than those with conidia, causing a greater negative impact on colony health. All types of baits containing Escovopsis weberi influenced the foraging activity but only treatments with viable fungal propagules resulted in an increase in the quantity of waste material, with a significant negative impact on the fungal garden biomass. The results provided novel information regarding Escovopsis recognition by worker ants and differences between conidia and mycelial pellet dynamics in leaf-cutting ant colonies, with new perspectives for the biological control of these important pests.

Islands in the shade: scattered ectomycorrhizal trees influence soil inoculum and heterospecific seedling response in a northeastern secondary forest.

The eastern deciduous forest is a mix of arbuscular (AM) and ectomycorrhizal (EM) trees, but land use legacies have increased the abundance of AM trees like Acer spp. (maple). Although these legacies have not changed the abundance of some EM trees like Betula spp. (birch), EM conifers like Tsuga canadensis (hemlock), and Pinus strobus (pine) have declined. We used a soil bioassay to investigate if the microbial community near EM birch (birch soil) contains a greater abundance and diversity of EM fungal propagules compatible with T. canadensis and P. strobus compared to the community associated with the surrounding AM-dominated secondary forest matrix (maple soil). We also tested the effectiveness of inoculation with soil from a nearby EM-dominated old-growth forest as a restoration tool to reintroduce EM fungi into secondary forest soils. Finally, we examined how seedling growth responded to EM fungi associated with each treatment. Seedlings grown with birch soil were colonized by EM fungi mostly absent from the surrounding maple forest. Hemlock seedlings grown with birch soil grew larger than hemlock seedlings grown with maple soil, but pine seedling growth did not differ with soil treatment. The addition of old-growth soil inoculum increased hemlock and pine growth in both soils. Our results found that EM trees are associated with beneficial EM fungi that are mostly absent from the surrounding AM-dominated secondary forest, but inoculation with old-growth soil is effective in promoting the growth of seedlings by reintroducing native EM fungi to the AM-dominated forests.

Biopolymer-based emulsions for the stabilization of Trichoderma atrobrunneum conidia for biological control

Trichoderma spp. are ubiquitous soil-borne fungi that are widely used in biological control to promote and regulate healthy plant growth, as well as protect against plant pathogens. However, as with many biological materials, the relative instability of Trichoderma propagules limits its practical use in industrial applications. Therefore, there has been significant research interest in developing novel formulations with various carrier substances that are compatible with these fungal propagules and can enhance the shelf-life and overall efficacy of the Trichoderma. To this end, herein, we investigate the use of a variety of biopolymers and nanoparticles for the stabilization of Trichoderma atrobrunneum T720 conidia for biological control. The best-performing agents—agar and cellulose nanocrystals (CNC)—were then used in the preparation of oil-in-water emulsions to encapsulate conidia of T720. Emulsion properties including oil type, oil:water ratio, and biopolymer/particle concentration were investigated with respect to emulsion stability, droplet size, and viability of T720 conidia over time. Overall, agar-based formulations yielded highly stable emulsions with small droplet sizes, showing no evidence of drastic creaming, or phase separation after 1 month of storage. Moreover, agar-based formulations were able to maintain ~ 100% conidial viability of T720 after 3 months of storage, and over 70% viability after 6 months. We anticipate that the results demonstrated herein will lead to a new generation of significantly improved formulations for practical biological control applications.Graphical abstractKey points• Various biopolymers were evaluated for improving the stability of Trichoderma conidia• Oil in water emulsions was prepared using cellulose nanocrystals and agar as interface stabilizers• Agar-based emulsions showed ~ 100% viability for encapsulated conidia after 3 months of storage

High concentration of spores and colony forming units of the biocontrol agent Beauveria bassiana via optimization of submerged cultivation

Liquid cultivation for the production of mycoinsecticides has received recent attention by the industry, due to the many advantages over the solid-state cultivation process. However, important variables must be considered to enable the production of the fungus propagules in the liquid medium. In this study, the optimization of the culture medium and the influence of pH and dissolved oxygen (DO) parameters on the production of spores of Beauveria bassiana in liquid process was investigated. The strain of B. bassiana IBCB 868 was growth in shake flasks followed by experimental design to find the optimal medium condition for spores production. Optimization of the culture medium was performed using a Plackett-Burman design followed by a central composite rotational design (CCRD). The optimized conditions resulted in high concentrations of submerged spores (2.21 × 109 SS mL−1) and colony forming units (1.52 × 109 CFU mL−1) on the third day of cultivation. There was an increase of 165.05% and 210.16% in SS and CFU production, respectively, in the optimized condition when compared to the most favorable culture medium initially tested. Benchtop bioreactor assays with the optimized medium were performed and the influence of pH and DO was investigated. It was observed that the pH fixed at 4.5 and 10% DO in 24 h, followed by 50% DO until 48 h and 30% DO until the end of the cultivation provided the highest concentrations of SS (1.98 × 109 SS mL−1) and CFU (1.9 × 109 CFU mL−1) after four days of growth.

Evaluation of the Richness, Species Diversity, and Biosafety of Saprotrophic Fungal Communities in Constructed Wetlands and Biological Wastewater Ponds

The present study was focused on the characterization of the biocenotic diversity of saprotrophic fungi, taking into account the impact of various treatment systems and technological stages as well as the physicochemical properties of wastewater. The biodegradation potential, the hazard posed to humans, animals, and the environment, and the effectiveness of elimination of propagules were evaluated. The most effective elimination of fungal propagules was recorded in hybrid constructed wetland systems with horizontal (HF-CW) and vertical (VF-CW) wastewater flow, especially in the VF-HF objects. The fungal communities present in wastewater from small constructed wetland (CW) and wastewater stabilization ponds (WSP) were dominated by ubiquitous terrestrial molds accompanied by a minimal number of yeasts. The similarity of the species composition of the fungal communities between the treatment plants was generally low, whereas the species diversity together with the population size was very high at the various stages of wastewater treatment. Species with potential pathogenicity to humans and animals accounted for over 45%, i.e., were classified as BioSafety Level 1 and 2 (BSL-1 and BSL-2 groups), and potentially phytopathogenic fungi represented 31.5% of the mycobiota species composition. The dynamics of fungal growth were correlated with the content of organic pollutants and nutrients (nitrogen and phosphorus) and with oxygen deficiency. The accumulation of nitrates corresponded to the decline in the frequency of fungi in treated wastewater. The lowest efficiency of the removal of fungi was exhibited by the biological wastewater stabilization ponds.

Omnivorous Notoxus trinotatus Pic (Coleoptera: Anthicidae) is a newly recognized vector of northern leaf blight in maize.

The adaptations of omnivorous insects to food are manifested in a multifaceted manner, and the availability of food resources directly determines insect feeding tendencies, which contribute to a complex insect-food relationship and impact insect functionality in the environment. Stable isotope analysis was applied to test the feeding preference and further define the functional role of omnivorous beetles in cropland. Our results confirmed that as an omnivorous beetle, the fungivorous nature of Notoxus trinotatus accounted for a prominent proportion food selection at the adult stage, and more importantly, this dietary feature contributed to the dispersal of the northern corn leaf blight in maize (NLB) during the feeding trials. In addition to the preference for fungi, water supplementation was an essential element extending adult longevity, which directly prolonged the contact time of adults with pathogenic fungi in agricultural fields. Consistent with the herbivorous characteristics of beetles, before the emergence of NLB fungal pathogens, corn tissues served as the main food, which provided the beetles with more opportunities to transmit fungal pathogen propagules. We conclude that the role of N. trinotatus in carrying NLB pathogen is due to its feeding on this plant mycopathogen, and an increased abundance of beetles carrying the pathogen may increase the rate of NLB disease infestation. More focus should be concentrated on the functions of fungivorous beetles, not only as pathogen-transmitting pests, but also as an element among the balanced biotic factors in farmland.

Three Methods Assessing the Association of the Endophytic Entomopathogenic Fungus Metarhizium robertsii with Non-Grafted Grapevine Vitis vinifera.

Characterizing the association of endophytic insect pathogenic fungi (EIPF) with plants is an important step in order to understand their ecology before using them in biological control programs. Since several methods are available, it is challenging to identify the most appropriate for such investigations. Here, we used two strains of Metarhizium robertsii: EF3.5(2) native to the French vineyard environment and ARSEF-2575-GFP a laboratory strain expressing a green fluorescent protein, to compare their potential of association with non-grafted grapevine Vitis vinifera. Three methods were used to evaluate the kinetics of rhizosphere and grapevine endospheric colonization: (i) Droplet Digital (ddPCR), a sensitive molecular method of M. robertsii DNA quantification in different plant parts, (ii) culture-based method to detect the live fungal propagules from plant tissues that grew on the medium, (iii) confocal imaging to observe roots segments. Both strains showed evidence of establishment in the rhizosphere of grapevines according to the culture-based and ddPCR methods, with a significantly higher establishment of strain EF3.5(2) (40% positive plants and quantified median of exp(4.61) c/μL) compared to strain ARSEF-2575-GFP (13% positive plants and quantified median of exp(2.25) c/μL) at 96-98 days post-inoculation. A low incidence of association of both strains in the grapevine root endosphere was found with no significant differences between strains and evaluation methods (15% positive plants inoculated with strain EF3.5(2) and 5% with strain ARSEF-2575-GFP according to culture-based method). ddPCR should be used more extensively to investigate the association between plants and EIPF but always accompanied with at least one method such as culture-based method or confocal microscopy.

Using sonic tomography to assess the relationship between internal wood decay and saproxylic beetle communities

Although wood-inhabiting beetle species are known to contribute to the progressive degradation of wood by building larval galleries and dispersing fungal propagules, the relationship between decaying wood and beetle communities is difficult to be assessed. The ecological properties of trees, such as their mortality status and species, could modify the relationships between beetle communities and decaying wood. We used recently developed techniques of sonic tomography for the first time to address how the trophic structure of beetle communities varies with the internal wood decay. We developed a paired design where sonic tomograms taken in the trunk sections of live and dead trees of the Patagonian forest were related with beetles sampled with emergence traps. Beetles captured in the trunks of live trees responded weakly to the internal wood decay. Indeed, the diversity of predators in live trees was likely influenced by bark microhabitats. Conversely, most of the guild assemblages in the trunks of dead trees were impoverished as wood decay was more advanced, possibly due to decreasing microhabitat quality. Our preliminary study suggests sonic tomography offers the opportunity to distinguish how beetles modulate decaying wood, but also provides insights about the assessment of live trees with tomograms. • Internal wood decay was evaluated with sonic tomography. • Wood decay and beetle communities were associated. • Beetles in live trees responded weakly to wood decay. • Beetles in dead trees responded negatively to wood decay.

Грибные болезни многолетних бобовых трав в Белгороде

We analyzed soil samples and symptoms of mycoses of red clover, goat's rue, alfalfa, birds-foot trefoil, sandy sainfoin in the phase of budding-beginning of flowering of plants against a natural infectious background in small-plot experiments in the territory of the botanical garden of the National Research University "BelSU" (Belgorod, RF). As a result, a series of decrease in the number of propagules of soil microscopic fungi (thousand CFU/g) was compiled: alfalfa (27.8 ± 4.01) – birds-foot trefoil (23.5 ± 2.25) – sainfoin (18.3 ± 1.67) – clover (17.3 ± 2.43) – goat's rue (13.7 ± 1.21) – fallow (12.3 ± 1.73). The number of micromycete species in the fallow soil was 38, and 9–22 species decreased in the grass rhizosphere. On red clover plants, powdery mildew was recorded with an intensity of development and spread of 64% and 75%, respectively, with a shortage of green mass productivity of more than 50%. The loss of green mass productivity in clover Alternariosis was 81%. Rust of alfalfa, birds-foot and sainfoin led to losses in the productivity of green mass up to 14% and was noted with a prevalence of 20%. Symptoms of purple rhizoctoniosis, fusarium root rot, black mold, downy mildew, and yellow spot were also recorded. More than 80% of phytopathogenic fungi are representatives of the leaf-stem group, the rest are of the root-tuber group.

ORAL MANIFESTATIONS OF PARACOCCIDIOIDOMYCOSIS: A CASE REPORT

Paracoccidioidomycosis is the main systemic mycosis in Brazil and the infection occurs by inhalation of fungus propagules. Several regions of the body can be affected, including the oral mucosa. The aim of this study is to relate a case of oral paracoccidioidomycosis. A 50-year-old male patient, residing in the countryside, had been hospitalized with symptoms of cough and fever and underwent treatment for tuberculosis in 2019. Computed tomography exam of the chest showed small nodular opacities and a "tree-in-bud" pattern. He had painful and itchy ulcerated lesions in the lips and tongue, in addition to the presence of granulomatous plaque with hemorrhagic stitches, for about 3 months. An incisional biopsy was performed, and the presence of fungi was confirmed by Grocott staining. Treatment with Bactrin was started in July 2020 and continues today. After 1 month of antibiotic treatment, the patient had a significant improvement and continues to be monitored.

Soil propagule banks of ectomycorrhizal fungi associated with Larix cajanderi above the treeline in the Siberian Arctic.

Microbial symbionts are essential for plant niche expansion into novel habitats. Dormant propagules of ectomycorrhizal (EM) fungi are thought to play an important role in seedling establishment in invasion fronts; however, propagule bank communities above the treeline are poorly understood in the Eurasian Arctic, where treelines are expected to advance under rapid climate change. To investigate the availability of EM fungal propagules, we collected 100 soil samples from Arctic tundra sites and applied bioassay experiments using Larix cajanderi as bait seedlings. We detected 11 EM fungal operational taxonomic units (OTUs) by obtaining entire ITS regions. Suillus clintonianus was the most frequently observed OTU, followed by Cenococcum geophilum and Sebacinales OTU1. Three Suillus and one Rhizopogon species were detected in the bioassay seedlings, indicating the availability of Larix-specific suilloid spores at least 30 km from the contemporary treeline. Spores of S. clintonianus and S. spectabilis remained infective after preservation for 14 mo and heat treatment at 60 °C, implying the durability of the spores. Long-distance dispersal capability and spore resistance to adverse conditions may represent ecological strategies employed by suilloid fungi to quickly associate with emerging seedlings of compatible hosts in treeless habitats.

Trends in Molecular Diagnostics and Genotyping Tools Applied for Emerging Sporothrix Species.

Sporotrichosis is the most important subcutaneous mycosis that affects humans and animals worldwide. The mycosis is caused after a traumatic inoculation of fungal propagules into the host and may follow an animal or environmental transmission route. The main culprits of sporotrichosis are thermodimorphic Sporothrix species embedded in a clinical clade, including S. brasiliensis, S. schenckii, S. globosa, and S. luriei. Although sporotrichosis occurs worldwide, the etiological agents are not evenly distributed, as exemplified by ongoing outbreaks in Brazil and China, caused by S. brasiliensis and S. globosa, respectively. The gold standard for diagnosing sporotrichosis has been the isolation of the fungus in vitro. However, with the advance in molecular techniques, molecular assays have complemented and gradually replaced the classical mycological tests to quickly and accurately detect and/or differentiate molecular siblings in Sporothrix. Nearly all techniques available for molecular diagnosis of sporotrichosis involve PCR amplification, which is currently moving towards detecting Sporothrix DNA directly from clinical samples in multiplex qPCR assays. From an epidemiological perspective, genotyping is key to tracing back sources of Sporothrix infections, detecting diversity in outbreak areas, and thus uncovering finer-scale epidemiological patterns. Over the past decades, molecular epidemiological studies have provided essential information to policymakers regarding outbreak management. From high-to-low throughput genotyping methods, MLSA, AFLP, SSR, RAPD, PCR-RFLP, and WGS are available to assess the transmission dynamics and sporotrichosis expansion. This review discusses the trends in the molecular diagnosis of sporotrichosis, genotyping techniques applied in molecular epidemiological studies, and perspectives for the near future.

Modeling Root-Knot Nematode Regulation by the Biocontrol Fungus Pochonia chlamydosporia.

Two models of increasing complexity were constructed to simulate the interactions between the root-knot nematode (RKN) Meloidogyne incognita and the biocontrol fungus Pochonia chlamydosporia var. catenulata in a rhizosphere microcosm. The models described discrete population dynamics at hourly rates over a 6-month period and were validated using real parasitism and nematode or fungus data. A first, general Pochonia-nematode-root model (GPNR) used five functions and 16 biological constants. The variables and constants describing the RKN life cycle included the rates of egg production, hatching, juvenile (J2), and mature female development, including root or nematode self-density-dependent factors. Other constants accounted for egg parasitism, nematode-induced root losses, growth, and mortalities. The relationship between nematodes and fungal propagules showed density dependence and cyclic variations in time, including an attractor on the propagules and J2 phases space. The simulations confirmed a P. chlamydosporia optimal initial density of 5 · 103 propagules · cc soil-1, as usually applied in assays. The constants used in GPNR showed adherence to the nematode biology, with 103 eggs per egg mass, a 10-day average lifespan of J2, with 2 days required to enter roots, and adult lifespan lasting 24 days. The fungus propagule lifespan was 25 days, with an average feeder root lifespan lasting around 52 days. A second, more complex Pochonia-nematode-root detailed model (GPNRd) was then constructed using eight functions and 23 constants. It was built as GPNR did not allow the evaluation of host prevalence. GPNRd allowed simulations of all RKN life stages and included non-parasitic and parasitic fungus population fractions. Both GPNR and GPNRd matched real J2 and fungus density data observed in a RKN biocontrol assay. Depending on the starting conditions, simulations showed stability in time, interpreted as effective host regulation. GPNRd showed a fungus cyclic relationship with the J2 numbers, with prevalence data close to those observed (38.3 vs. 39.4%, respectively). This model also showed a further density-independent nematode regulation mechanism based on the P. chlamydosporia switch from a non-parasitic to a parasitic trophic behavior. This mechanism supported the biocontrol of M. incognita, also sustained by a concomitant increase of the root density.

Indoor air quality and diversity of fungi inside and outside residences of children with a history of allergy in Cuba

Several investigations suggest that the exposure to fungal allergens during childhood may increase the risk of sensitisation in children genetically predisposed to allergy. The objective of this study was to evaluate the quality and diversity of fungal propagules in indoor air, specifically in the bedrooms of children with a family history of allergy. The indoor and outdoor air was sampled from 44 bedrooms of children with a family history of allergy during the years 2018 and 2019 in Havana, Cuba. Inside the bedrooms, the presence of humidity problems, visible fungal growth, and the quality of ventilation was examined, while the values of temperature and relative humidity were recorded. Moreover, a distribution analysis of the genera detected was carried out and the Sørensen coefficient of similarity was calculated. In addition, the relationship between the outdoor and indoor air concentrations of each residence was determined. The concentrations of fungal propagules in the indoor air of the bedrooms were between 20 and 1330 colony-forming unit (CFU)/m3. According to the analysed limit values, 18 bedrooms can be classified as having poor indoor air quality. Cladosporium, Aspergillus, Penicillium, and Curvularia were the most frequent genera of the 19 identified in the study. There was a similarity between the indoor and outdoor air mycobiota in 15.9% of the rooms. The detection of these allergenic fungal genera is an alert for children in the sampled homes, mainly for those that were classified as poor indoor air quality according to the examined standards.

Current developments in the resistance, quality, and production of entomopathogenic fungi.

There is a worldwide concern to achieve food security with a sustainable approach, including the generation and implementation of techniques for the production of high-quality chemical-free crops. This food revolution has promoted the development and consolidation of programmes for integrated pest management. Some of those programmes include the use of diverse organisms (biological control agents) to suppress populations of pests potentially harmful to the crops. Among these biological control agents are entomopathogenic fungi that are highly effective in suppressing a diversity of insects and have, therefore, been produced and marketed throughout the world. However, the bottleneck for applying entomopathogenic fungi is the production of propagules (blastospores and conidia) with resistance to environment conditions and abiotic factors, maintaining high quality in terms of virulence. Therefore, this manuscript presents recent studies related to increasing resistance and quality using different bioreactors to produce conidia. The above presents a global panorama related to current developments that contribute to improving the resistance, quality, and production of entomopathogenic fungal propagules.

Clonostachys rosea: Production by Submerged Culture and Bioactivity Against Sclerotinia sclerotiorum and Bemisia tabaci.

Among the prospective biocontrol agents, the saprophytic filamentous fungus Clonostachys rosea is an excellent necrotrophic mycoparasite of numerous plant pathogenic fungi. However, its commercial development has been hampered by mass production difficulties during solid-state fermentation. Conversely, the submerged liquid fermentation shortens the cultivation time while increasing yields of fungal propagules. However, this method has been overlooked for C. rosea. In this work, we investigated the impact of liquid pre-culture inoculum on the spore production by the two-stage fermentation process using rice grains in comparison to the traditional solid-state fermentation. In parallel, we studied the submerged cultivation of C. rosea by manipulating carbon-to-nitrogen (C:N) ratio and nitrogen source, with the further optimization of spore production in a benchtop bioreactor. Additional bioassays included assessing the bioactivity of water-dispersible microgranules (that contained a submerged conidia) against the whitefly (Bemisia tabaci biotype B) and Sclerotinia sclerotiorum (causal agent of the white mold). Our results showed a maximum concentration of 1.1 × 109 conidia/g-dry-matter after 7 days of cultivation by two-stage fermentation process. The liquid fermentation yielded 1.4 × 109 submerged conidia/ml after 7 days using a medium with a 50:1 C:N ratio, and it also induced the production of microsclerotia (MS) up to 1.35 × 104/ml within 6 days with 10:1 C:N ratio; both media were supplemented with dextrose monohydrate and soybean meal. The fermentation batches carried out in a benchtop bioreactor with medium 50:1 C:N ratio and amended with soybean meal rendered a production peak on the fourth day, corresponding to 1.11 × 109 conidia/ml and 4.35 × 108 colony forming units (CFU)/ml. Following air-drying, the conidia production from air-dried microgranules of C. rosea biomass was estimated at 3.4 × 1010 conidia/g of formulated product upon re-hydration for 7 days. Both submerged conidia and MS of C. rosea inhibited 100% germination of S. sclerotiorum sclerotia by direct parasitism. The air-dried submerged conidia exhibited a suppressive activity on sclerotia (88% mycoparasitism) and early whitefly nymphs (76.2% mortality) that rendered LC50 values of 3.2 × 104 CFU/g soil and 1.5 × 107 CFU/ml, respectively. Therefore, the submerged liquid culture of C. rosea may offer a feasible and cost-effective method for its large-scale production, alleviating critical constraints to their commercial use while providing an additional tool for management of B. tabaci and S. sclerotiorum.

Unselective Transport of Phytopathogenic Fusarium Fungi from Litter and Soil by Ground-Dwelling Arthropods Links Semi-Natural and Agricultural Habitats.

The dispersal of propagules, such as fungal spores or seeds by actively moving animals, connects and shapes communities. The dispersal of plant pathogens by arthropods might be a crucial mechanism in the spread of several crop diseases. Ground-dwelling arthropods are potential linkers between fungal communities in semi-natural and agricultural habitats by transporting propagules of Fusarium fungi. We compared the Fusarium communities on the body surface of ground-dwelling arthropods with litter in semi-natural and soil in agricultural habitats with a focus on the Fusarium community. We found three relatively distinct Fusarium communities with moderate overlap. We detected a higher richness of Fusarium species on the body surface of arthropods compared to litter and soil communities. The results suggest that the Fusarium community on the body surface of arthropods relates to the composition observed in litter and soil with limited filtering mechanisms between communities. Ground-dwelling arthropods are relevant agents for the distribution of Fusarium and therefore link fungal communities in adjacent habitats.