Use of natural-based commercial products as an alternative for providing bioprotection against verticillium wilt of olive.

BACKGROUND: As a result of the ineffectiveness of existing control methods against Verticillium dahliae, the causal agent of verticillium wilt of olive (Olea europaea; VWO), it is necessary to search for sustainable and environmentally friendly alternatives, such as bioprotection by products based on plant extracts and other naturally synthesized compounds. Therefore, present study aimed to evaluate the effects of seven natural-based commercial products on the inhibition of mycelial growth, the germination of V. dahliae conidia and microsclerotia, and disease progression in olive plants (cv. Picual). Aluminium lignosulfonate and a copper phosphonate salt (copper phosphite) were included for comparative purposes. RESULTS: The seaweed and willow extracts and copper phosphite inhibited V. dahliae mycelial growth by more than 50% at the high doses tested. Most of the products inhibited conidial germination by up to 90% compared to the control at the high doses tested. However, none of the products showed efficacy above 50% in inhibiting microsclerotia germination. The willow extract was the most effective at reducing disease severity and progression in olive plants, with no significant differences compared to the non-inoculated negative control. CONCLUSION: The results of the present study suggest that the use of natural-based products (i.e. seaweed and willow extracts) is a potential sustainable alternative in an integrated VWO control strategy. © 2024 Society of Chemical Industry.

Dipeptidase PEPDA Is Required for the Conidiation Pattern Shift in Metarhizium acridum.

Filamentous fungi conduct two types of conidiation, typical conidiation from mycelia and microcycle conidiation (MC). Fungal conidiation can shift between the two patterns, which involves a large number of genes in the regulation of this process. In this study, we investigated the role of a dipeptidase gene pepdA in conidiation pattern shift in Metarhizium acridum, which is upregulated in MC pattern compared to typical conidiation. Results showed that disruption of the pepdA resulted in a shift of conidiation pattern from MC to typical conidiation. Metabolomic analyses of amino acids showed that the levels of 19 amino acids significantly changed in ΔpepdA mutant. The defect of MC in ΔpepdA can be rescued when nonpolar amino acids, α-alanine, ß-alanine, or proline, were added into sucrose yeast extract agar (SYA) medium. Digital gene expression profiling analysis revealed that PEPDA mediated transcription of sets of genes which were involved in hyphal growth and development, sporulation, cell division, and amino acid metabolism. Our results demonstrated that PEPDA played important roles in the regulation of MC by manipulating the levels of amino acids in M. acridum. IMPORTANCE Conidia, as the asexual propagules in many fungi, are the start and end of the fungal life cycle. In entomopathogenic fungi, conidia are the infective form essential for their pathogenicity. Filamentous fungi conduct two types of conidiation, typical conidiation from mycelia and microcycle conidiation. The mechanisms of the shift between the two conidiation patterns remain to be elucidated. In this study, we demonstrated that the dipeptidase PEPDA, a key enzyme from the insect-pathogenic fungus Metarhizium acridum for the hydrolysis of dipeptides, is associated with a shift of conidiation pattern. The conidiation pattern of the ΔpepdA mutant was restored when supplemented with the nonpolar amino acids rather than polar amino acids. Therefore, this report highlights that the dipeptidase PEPDA regulates MC by manipulating the levels of amino acids in M. acridum.

Functional Analysis of Autophagy-Related Gene ATG12 in Potato Dry Rot Fungus Fusarium oxysporum.

Autophagy is an intracellular process in all eukaryotes which is responsible for the degradation of cytoplasmic constituents, recycling of organelles, and recycling of proteins. It is an important cellular process responsible for the effective virulence of several pathogenic plant fungal strains, having critical impacts on important crop plants including potatoes. However, the detailed physiological mechanisms of autophagy involved in the infection biology of soil-borne pathogens in the potato crop needs to be investigated further. In this study, the autophagy-related gene, FoATG12, in potato dry rot fungus Fusarium oxysporum was investigated by means of target gene replacement and overexpression. The deletion mutant ∆FoATG12 showed reduction in conidial formation and exhibited impaired aerial hyphae. The FoATG12 affected the expression of genes involved in pathogenicity and vegetative growth, as well as on morphology features of the colony under stressors. It was found that the disease symptoms were delayed upon being inoculated by the deletion mutant of FoATG12 compared to the wild-type (WT) and overexpression (OE), while the deletion mutant showed the disease symptoms on tomato plants. The results confirmed the significant role of the autophagy-related ATG12 gene in the production of aerial hyphae and the effective virulence of F. oxysporum in the potato crop. The current findings provid an enhanced gene-level understanding of the autophagy-related virulence of F. oxysporum, which could be helpful in pathogen control research and could have vital impacts on the potato crop.

High voltage atmospheric cold plasma treatment inactivates Aspergillus flavus spores and deoxynivalenol toxin.

Fungal contamination is a concern for the food industry. Fungal spores resist food sterilization treatments and produce mycotoxins that are toxic for animals and humans. Technologies that deactivate spores and toxins without impacting food quality are desirable. This study demonstrates the efficiency of a high voltage atmospheric cold plasma (HVACP) technology using air to generate reactive oxygen (ROS) and nitrogen (RNS) species for the degradation of Aspergillus flavus cultures and the deoxynivalenol (DON) mycotoxin. Optical emission and absorption spectroscopy demonstrate ionization of hydroxyl groups, atomic oxygen and nitrogen, and confirm production of ROS and RNS, e.g. O3, NO2, NO3, N2O4, and N2O5. Fungal cultures show a depletion in pigmentation and an ~50% spore inactivation after 1-min treatments. Treated spores show surface ablation and membrane degradation by scanning electron microscopy. Twenty-minute direct HVACP treatments of 100 µg of DON in one mL aqueous suspensions resulted in a greater than 99% reduction in DON structure and rescued over 80% of Caco-2 cell viability; however, the same treatment on 100 µg of powdered DON toxin only showed a 33% reduction in DON and only rescued 15% of cell viability. In summary, HVACP air treatment can inactivate both fungal spores and toxins in minutes.

Aspergillus spp. eliminate Sclerotinia sclerotiorum by imbalancing the ambient oxalic acid concentration and parasitizing its sclerotia.

Sclerotinia sclerotiorum, a pathogen of more than 600 host plants, secretes oxalic acid to regulate the ambient acidity and provide conducive environment for pathogenicity and reproduction. Few Aspergillus spp. were previously proposed as potential biocontrol agents for S. sclerotiorum as they deteriorate sclerotia and prevent pathogen's overwintering and initial infections. We studied the nature of physical and biochemical interactions between Aspergillus and Sclerotinia. Aspergillus species inhibited sclerotial germination as they colonized its rind layer. However, Aspergillus-infested sclerotia remain solid and viable for vegetative and carpogenic germination, indicating that Aspergillus infestation is superficial. Aspergillus spp. of section Nigri (Aspergillus japonicus and Aspergillus niger) were also capable of suppressing sclerotial formation by S. sclerotiorum on agar plates. Their culture filtrate contained high levels of oxalic, citric and glutaric acids comparing to the other Aspergillus spp. tested. Exogenous supplementation of oxalic acid altered growth and reproduction of S. sclerotiorum at low concentrations. Inhibitory concentrations of oxalic acid displayed lower pH values comparing to their parallel concentrations of other organic acids. Thus, S. sclerotiorum growth and reproduction are sensitive to the ambient oxalic acid fluctuations and the environmental acidity. Together, Aspergillus species parasitize colonies of Sclerotinia and prevent sclerotial formation through their acidic secretions.

Trait-based approaches reveal fungal adaptations to nutrient-limiting conditions.

The dependency of microbial activity on nutrient availability in soil is only partly understood, but highly relevant for nutrient cycling dynamics. In order to achieve more insight on microbial adaptations to nutrient limiting conditions, precise physiological knowledge is needed. Therefore, we developed an experimental system assessing traits of 16 saprobic fungal isolates in nitrogen (N) limited conditions. We tested the hypotheses that (1) fungal traits are negatively affected by N deficiency to a similar extent and (2) fungal isolates respond in a phylogenetically conserved fashion. Indeed, mycelial density, spore production and fungal activity (respiration and enzymatic activity) responded similarly to limiting conditions by an overall linear decrease. By contrast, mycelial extension and hyphal elongation peaked at lowest N supply (C:N 200), causing maximal biomass production at intermediate N contents. Optimal N supply rates differed among isolates, but only the extent of growth reduction was phylogenetically conserved. In conclusion, growth responses appeared as a switch from explorative growth in low nutrient conditions to exploitative growth in nutrient-rich patches, as also supported by responses to phosphorus and carbon limitations. This detailed trait-based pattern will not only improve fungal growth models, but also may facilitate interpretations of microbial responses observed in field studies.

The deubiquitinating enzyme MoUbp8 is required for infection-related development, pathogenicity, and carbon catabolite repression in Magnaporthe oryzae.

Deubiquitination is an essential regulatory step in the Ub-dependent pathway. Deubiquitinating enzymes (DUBs) mediate the removal of ubiquitin moieties from substrate proteins, which are involved in many regulatory mechanisms. As a component of the DUB module (Ubp8/Sgf11/Sus1/Sgf73) in the SAGA (Spt-Ada-Gcn5-acetyltransferase) complex, Ubp8 plays a crucial role in both Saccharomyces cerevisiae and humans. In S. cerevisiae, Ubp8-mediated deubiquitination regulates transcriptional activation processes. To investigate the contributions of Ubp8 to physiological and pathological development of filamentous fungi, we generated the deletion mutant of ortholog MoUBP8 (MGG-03527) in Magnaporthe oryzae (syn. Pyricularia oryzae). The ΔMoubp8 strain showed reduced sporulation, pathogenicity, and resistance to distinct stresses. Even though the conidia of the ΔMoubp8 mutant were delayed in appressorium formation, the normal and abnormal (none-septum or one-septum) conidia could finally form appressoria. Reduced melanin in the ΔMoubp8 mutant is highly responsible for the attenuated pathogenicity since the appressoria of the ΔMoubp8 mutant was much more fragile than those of the wild type, due to the defective turgidity. The weakened ability to detoxify or scavenge host-derived reactive oxygen species (ROS) further restricted the invasion of the pathogen. We also showed that carbon derepression, on the one hand, rendered the ΔMoubp8 strain highly sensitive to allyl alcohol, on the other hand, it enhances the resistance of the MoUBP8 defective strain to deoxyglucose. Overall, we suggest that MoUbp8 is not only required for sporulation, melanin formation, appressoria development, and pathogenicity but also involved in carbon catabolite repression of M. oryzae.

Transcriptome Analysis Reveals the Effects of Chinese Chive (Allium tuberosum R.) Extract on Fusarium oxysporum f. sp. radicis-lycopersici Spore Germination.

Fusarium oxysporum f. sp. radicis-lycopersici (Forl) causes Fusarium crown and root rot of tomato, leading to severe yield losses. Chinese chive and the Chinese chive extract reportedly have antifungal effects. In this study, Chinese chive extract treatments inhibited Forl spore germination, with an EC50 of 0.40 g ml-1 in vitro. Furthermore, the mechanism underlying the fungicidal effects of the Chinese chive extract was analyzed by RNA sequencing. A total of 1252 differentially expressed genes (DEGs) were detected, of which 396 were upregulated and 856 were downregulated. The DEGs were related to starch and sucrose metabolism, amino sugar and nucleotide sugar metabolism, galactose metabolism, fatty acid metabolism, sphingolipid metabolism, glycerophospholipid metabolism, peroxisomes, ribosome biogenesis in eukaryotes, mismatch repair, and the phosphatidylinositol signaling system, implying these pathways contribute to the fungicidal activity of the Chinese chive extract. The qRT-PCR results verified the accuracy of the RNA sequencing data. Thus, the Chinese chive extract can inhibit Forl spore germination by affecting spore nutrient metabolism.

The role of in vitro cultivation on symbiotic trait and function variation in a single species of arbuscular mycorrhizal fungus.

In vitro propagation of AM fungi using transformed root cultures (TRC) is commonly used to obtain pure AM fungal propagules for use in research and industry. Early observations indicate that such an artificial environment can alter traits and function of AM fungi over time. We hypothesized that increased in vitro cultivation may promote ruderal strategies in fungi by enhancing propagule production and reducing mutualistic quality. To examine the effect of in vitro cultivation on the trait and function of AM fungi, we inoculated plants with 11 Rhizoglomus irregulare isolates which fell along a cultivation gradient spanning 80 generations. We harvested plants at 10, 20 and 30 d post inoculation to observe differences in fungal and plant traits post infection. In vitro cultivation led to increased spore production but reduced plant shoot phosphorus. Our results indicate that in vitro propagation may indirectly select for traits that affect symbiotic quality.

Plant-soil feedback of two legume species in semi-arid Brazil.

Positive feedback between arbuscular mycorrhizal fungal (AMF) and vascular plants can contribute to plant species establishment, but how this feedback affects plant invasion by Prosopis juliflora SW. (DC.), or resistance to invasion by Mimosa tenuiflora (Willd.) Poir in Brazilian semi-arid region is not well known. In this work, we tested how modified and native AMF communities affect the establishment of P. juliflora and M. tenuiflora plants. We examined the effects of inoculation with modified and native AMF communities on number of AMF spores, root colonization, number of N-fixing nodules, plant dry biomass, plant phosphorous concentration, and plant responsiveness to mycorrhizas of P. juliflora and M. tenuiflora. We found that the modified AMF community enhanced the root colonization, plant dry biomass, and plant phosphorous concentration of invasive P. juliflora, whereas native AMF enhanced M. tenuiflora. Our results demonstrate that the invasive P. juliflora alters soil AMF community composition, and this change generates positive feedback to the invasive P. juliflora itself and decreases AMF associations with native M. tenuiflora.

Inhibition of Alternaria stem canker on tomato by essential oils from Baccharis species.

The toxicity of four essential oils extracted from Baccharis articulata, Baccharis ochracea, Baccharis psiadioides and Baccharis trimera was tested against the phytopathogen Alternaria alternata, which causes Alternaria stem canker on tomatoes. Diseases caused by Alternaria fungi are responsible for great economic losses in terms of production and are controlled by synthetic fungicides; however, essential oils offer an alternative, since they have been proven to be effective for controlling against various plant pathogens. In this way, the antifungal activity of Baccharis essential oils was tested using potato dextrose agar medium with concentrations ranging from 0.1 to 20.0 µL mL-1. Baccharis trimera and Baccharis ochracea essential oils presented 100% mycelial growth inhibition of A. alternata and were also able to control Alternaria stem canker disease under greenhouse conditions. Tomato plants treated with these essential oils exhibited area under the disease progress curve (AUDPC) values of 230.10 and 241.42, differing from the control condition, which showed an AUDPC value of 268.92. The essential oils of B. trimera and B. ochracea can be an alternative for controlling Alternaria stem canker disease of tomatoes and should be formulated as a potential fungicide against the A. alternata pathogen.

The new species Puccinia telimutans causes Mexican potato rust disease of wild potatoes and shows a unique telial metamorphosis.

Puccinia telimutans is described as a new species of rust fungi (Basidiomycota, Pucciniales) and causal agent of Mexican potato rust disease of the endemic wild potatoes Solanum demissum and S. verrucosum from Mexico. It is microcyclic and produces telia that show a unique succession of one-celled, catenulate teliospores that germinate upon maturity, followed by resting teliospores that are pedicellate and two-celled. Puccinia telimutans appears to be restricted to Mexico and has formerly been confused with P. pittieriana, which causes common potato rust in South America.

Activity of additives and their effect in formulations of Metarhizium anisopliae s.l. IP 46 against Aedes aegypti adults and on post mortem conidiogenesis.

BACKGROUND: Oil formulations of entomopathogenic fungi have interest for biological mosquito control. OBJECTIVES: The activities of M. anisopliae s.l. IP 46 conidia were tested in Aedes aegypti adults either without any formulation or formulated with vegetable or mineral oil and in combination with diatomaceous earth. FINDINGS: IP 46 was highly active against adults, the vector of important arboviruses in the tropics and subtropics. At an exposure of adults to 3.3 × 107 conidia/cm2, values of lethal times TL50 and TL90 reached minimal 3.8 and 4.6 days, respectively, and lethal concentrations LC50 and LC90 were 2.7 × 105 and 2.4 × 106 conidia/cm2, respectively, after 10 days of exposure. Activity against adults was improved by diatomaceous earth (KeepDry® KD) combined with mineral oil (Naturol® N) or vegetable oil (Graxol® G). Additives KD or N separately (and G to a lesser extent) or in combination, KD + N and KD + G without conidia had also a clear adulticidal effect. Efficacy of conidia formulated or not with KD + N decreased somewhat at shorter exposure periods. Time of exposure (0.017, 12, 48, 72 or 120 h) of adults to KD and N or IP 46 or conidia and KD and N had no significant effect on mortality. M. anisopliae s.l. recycled on fungus-killed mosquitoes producing high quantities of new conidia regardless of the conidial concentrations or formulations tested. Additives tested had no clear effect on quantitative conidiogenesis on cadavers. MAIN CONCLUSIONS: Formulations of IP 46 conidia with mineral oil and diatomaceous earth represent a promising tool for the development of potent strategies of focal control of this important vector with entomopathogenic fungi.

Comparative metabolomics of scab-resistant and susceptible apple cell cultures in response to scab fungus elicitor treatment.

Apple scab disease caused by the fungus Venturia inaequalis is a devastating disease that seriously affects quality and yield of apples. In order to understand the mechanisms involved in scab resistance, we performed gas chromatography-mass spectrometry based metabolomics analysis of the cell culture of scab resistant cultivar 'Florina' and scab susceptible cultivar 'Vista Bella' both prior -to and -following treatment with V. inaequalis elicitor (VIE). A total 21 metabolites were identified to be altered significantly in 'Florina' cell cultures upon VIE-treatment. Among 21 metabolites, formation of three new specialized metabolites aucuparin, noraucuparin and eriobofuran were observed only in resistant cultivar 'Florina' after the elicitor treatment. The score plots of principal component analysis (PCA) exhibited clear discrimination between untreated and VIE-treated samples. The alteration in metabolite levels correlated well with the changes in the transcript levels of selected secondary metabolite biosynthesis genes. Aucuparin, noraucuparin and eriobofuran isolated from the 'Florina' cultures showed significant inhibitory effect on the conidial germination of V. inaequalis. The results expand our understanding of the metabolic basis of scab-resistance in apple and therefore are of interest in apple breeding programs to fortify scab resistance potential of commercially grown apple cultivars.

Determination of total phenolic content, antioxidant activity and antifungal effects of Thymus vulgaris, Trachyspermum ammi and Trigonella foenum-graecum extracts on growth of Fusarium solani.

Ajowan, thyme and fenugreek are spice and aromatic crops with a number of medicinal properties which are known as important sources of essential phytochemicals. The purpose of this study was to investigate the antioxidant capacity, total phenolic content and antifungal activities of these plant extracts on growth of Fusarium solani, an important plant pathogen, soil saprophyte and one of the causal agents of fusariosis in human and animals. Their total antioxidant activity was measured using DPPH radical scavenging assay and their antimicrobial activity was determined through poison food assay at two concentrations (1000 and 1500 ppm) and spore germination assay in vitro. All methanolic extracts showed high antioxidant activity which among them methanolic extract of thyme demonstrated higher antioxidant potential with a low IC50 (16.50 mg ascorbic acid/g). Also, the highest phenolic content (70.55 mg GAE g-1) was observed in methanolic extract of thyme. The highest and lowest amount of thymol was determined in methanolic extract of thyme and aqueous extract of ajowan. Methanolic extracts of thyme leaves and ajowan seeds at concentration of 1500 ppm were potentially effective against F. solani over the control treatments by 90.33% and 85.73%, respectively. Followed by hydro-ethanolic and aqueous extracts exhibited a lesser percentage of inhibition. The MIC value for methanolic extract of thyme and ajowan was 3.75 mg/ml followed by hydro-ethanolic and aqueous extracts, respectively. The amount of calculated MFC was ranging from 7.5 to 30 mg/ml for thyme methanolic and fenugreek aqueous extracts, respectively.

Existence Arbuscula Mycorrhiza and Its Application Effect to Several Variety of Corn Plant (Zeal mays L.) in Marginal Dry Land.

BACKGROUND AND OBJECTIVE: Arbuscula mycorrhiza (AM) have a very large function in symbiosis with plant roots, it's very important to be studied further because AM utilization is an alternative solution to improve the yield of corn plant in poor land. Until now the productivity of corn plant, especially in Southeast Sulawesi, Indonesia is lower than it's genetic potential, one of the causes is the cultivation of many plant done in sub optimal land with low technology applications especially the use of organic and biological fertilizer very low. This study aimed to observe the presence of AM and evaluate the growth and productivity of corn plant that AM inoculated. MATERIALS AND METHODS: The study consisted two series of experiments, namely (1) Existence arbuscula mycorrhiza in rhizosphere of dominant weed (bladygrass) in dry land in Kendari, Southeast Sulawesi, Indonesia, was observed descriptively, (2) Effect of arbuscula mycorrhiza to several variety of corn plant (Zea mays L.) in Marginal dry land. The experiment was arranged based Randomized Block Design (RBD) consisted of eight treatments corn variety. RESULTS: The results showed that spore populations were found in rhizosphere of dominant weed was 792-901 spores 100 g-1 soil, 70-90% roots infection. Some genera found are Glomus, Gigasphora, Acaulospora, Entrophospora, Scutellospora. Symbiosis effect of arbuscula mycorrhiza with plant growth indicates that Phosphorus uptake was highest in Batu Putih variety. However the highest in yield was show in Dana variety. Compared with the lowest production, the production difference was higher in Dana 47.70%. CONCLUSION: Existence of arbuscula mycorrhiza in dominant weed rhizosphere in dry land is very high. The response of various varieties of maize plants to arbuscula mycorrhiza indicates that local varieties have a higher adaptability compared with introduction varieties.

Interactive effects of dissolved nitrogen, phosphorus and litter chemistry on stream fungal decomposers.

The enrichment of ecosystems by nutrients such as nitrogen (N) and phosphorus (P) has important ecological consequences. These include effects on plant litter decomposition in forest soils and forested headwater streams, where fungi play a pivotal role. However, our understanding of nutrient relationships on fungal communities associated with decomposing litter remains surprisingly incomplete. We conducted a fully factorial microcosm experiment with known communities of fungal decomposers from streams to assess the importance of dissolved N and P supply, as well as the atomic nutrient ratio (N:P), on fungal community succession, diversity, biomass and reproduction on three leaf-litter species differing in nutrient and lignin concentrations. Fungal biomass accrual and spore production were strongly controlled by external N supply, whereas P supply was much less important. The magnitude of these effects was mediated by litter quality, with stronger effects of dissolved N and P on lignin-poor and high N:P litter. N supply also influenced fungal diversity and species composition, acting as a pacemaker of community succession. Collectively, our data indicate that N was in much greater demand than predicted by standard stoichiometric models. The most parsimonious explanation for this deviation relates to the need of litter fungi to invest large amounts of N into degradative exoenzymes.

Morphological Characteristics of Monokaryotic and Dikaryotic Collections of Three Medicinal Coprinellus Species (Agaricomycetes).

This article presents data from morphological observations of mycelia of 40 monokaryotic and 11 dikaryotic collections of 3 medicinal Coprinellus species (C. disseminatus, C. micaceus, and C. xanthothrix). The growth rate, colony morphology, and micromorphological characteristics of mycelia and anamorphs on 1.5% malt-extract agar (MEA) and potato-dextrose agar (PDA) are described. Well-developed white, cottony-felt colonies, which later show creamy, yellowish to rusty brown pigmentation on mycelia and agar, were typical for the studied Coprinellus collections. Mycelial growth was denser on PDA than on MEA, whereas the average growth rate indicators (GRavr) were higher in dikaryotic isolates on MEA. Clamp connections were described only in dikaryotic isolates of C. disseminatus and C. micaceus; mycelia of C. xanthothrix had no clamps. Nonsporulating Ozonium-type anamorphic mycelia (with a rusty brown septate and parallel hyphal strands), a taxonomic feature characteristic of the clade Coprinellus, was present in the studied monokaryotic and dikaryotic collections, whereas Hormographiella-type sporulating anamorphs developed only in monokaryotic and dikaryotic isolates of C. xanthothrix. Yellowish-rusty-brownish regular hyphal loops were also observed in the collections of all 3 Coprinellus species. Allocyst-like hyphal swellings were observed in monokaryons of C. xanthothrix, and hyphocystidia were observed in dikaryons of C. micaceus. Hyphal loops and hyphal cystidia presumably were derived from Ozonium mycelia. Thick-walled, oval chlamydospores and chlamydospore-like swellings were described only in dikaryons of C. xanthothrix. Under these experimental conditions, primordia and fruiting bodies developed in dikaryons of C. xanthothrix on MEA and PDA, respectively, and in dikaryons of C. micaceus on MEA. The taxonomic significance of the mycelial and anamorphic characteristics of studied Coprinellus species was evaluated. They could be useful for identifying mycelial cultures during biotechnological cultivation.

Inter- and intra-species variability in heat resistance and the effect of heat treatment intensity on subsequent growth of Byssochlamys fulva and Byssochlamys nivea.

The major aims of this study were to assess inter- and intra-species variability of heat resistant moulds (HRMs), Byssochlamys fulva and Byssochlamys nivea, with regards to (i) heat resistance and (ii) effect of heat treatment intensity on subsequent outgrowth. Four-week-old ascospores were suspended in buffered glucose solution (13° Brix, pH 3.5) and heat treated in a thermal cycler adjusted at 85 °C, 90 °C and 93 °C. Two variants of the Weibull model were fitted to the survival data and the following inactivation parameters estimated: b (inactivation rate, min-1), n (curve shape) and δ (the time taken for first decimal reduction, min). In addition to the assessment of heat resistance, outgrowth of Byssochlamys sp. from ascospores heated at 70 °C, 75 °C, 80 °C, 85 °C and 90 °C for 10 min and at 93 °C for 30 and 70 s was determined at 22 °C for up to 30 days. The Baranyi and Roberts model was fitted to the growth data to estimate the radial growth rates (µmax, mm.day-1) and lag times (λ, days). Inter-species variability and significant differences (p < 0.05) were observed for both inactivation and growth estimated parameters among B. fulva and B. nivea strains. The effect of heat treatment intensity on outgrowth of B. fulva strains was more apparent at the most intense heat treatment evaluated (90 °C/10 min), which was also the condition in which greater dispersion of the estimated kinetic parameters was observed. On the other hand, B. nivea strains were more affected by heating, resulting in greater variability of growth parameters estimated at different heating intensities and in very long lag phases (up to 25 days). The results show that inter- and intra-species variability in the kinetic parameters of Byssochlamys sp. needs to be taken into account for more accurate spoilage prediction. Furthermore, the effect of thermal treatments on subsequent outgrowth from ascospores should be explored in combination with other relevant factors such as °Brix and oxygen to develop thermal processes and storage conditions which can prevent the growth of HRMs and spoilage of heat treated food products.

Cellulase enhances endophytism of encapsulated Metarhizium brunneum in potato plants.

The recent discovery that entomopathogenic fungi can grow endophytically in plant tissues has spurred research into novel plant protection measures. However, current applications of fungi aiming at endophytism mostly lack targeted formulation strategies resulting in low efficacy. Here, we aimed at enhancing Metarhizium brunneum CB15 endophytism in potato plants by (i) improvement of fungal growth from beads and (ii) cellulase formation or addition to encapsulated mycelium. We found that beads supplemented with cellulose alone or in addition with inactivated baker's yeast exhibited cellulase activity and increased mycelial growth by 12.6 % and 13.6 %, respectively. Higher enzymatic activity achieved by cellulase co-encapsulation promoted a shift from mycelial growth to spore formation with maximum numbers of 2.5 × 108 ± 6.1 × 107 per bead. This correlated with improved endophytism in potato plants by 61.2 % compared to non-supplemented beads. Our study provides first evidence that customized formulations of fungal entomopathogens with enzymes can improve endophytism and this may increase efficacy in plant protection strategies against herbivorous pests.