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The use of antagonistic microorganisms in the suppression of phytopathogenic fungi represents the ultimate approach to biological control, and because of their significant beneficial effects, antagonistic microorganisms are increasingly being researched. The necrotrophic fungus Botrytis cinerea known as the causative agent of gray mold, leads to significant global yield and post-harvest storage losses. It is a polyphagous of high risk for fungicide resistance development, which is why the use of antagonistic microorganisms a promising biocontrol strategy. Among the antagonistic microorganisms, the entomopathogenic fungus Beauveria bassiana is considered to have antagonistic properties in controlling phytopathogenic fungi due to the wide range of secondary metabolites. In order to maximize the antagonistic fungi effectiveness in field conditions, special attention paid to researching the antagonistic mechanisms under optimal conditions for their development. The aim of this study is testing antifungal effect of B. bassiana on pathogen B. cinerea by dual cultures method and testing a production of volatile metabolites on a substrate with two different pH values (pH= 5.6 and 7.2). The results of dual culture method and testing a production of volatile metabolites confirmed that the antagonistic effect of B. bassiana is significantly higher on a neutral pH substrate, where the inhibition of the B. cinerea was 36%, i.e. 38% which was achieved by different antagonistic mechanisms. Microscopic analysis confirmed inhibition mechanisms (mycoparasitism and antibiosis), more significant on a neutral pH substrate by B. bassiana, and B. cinerea microstructures deformations. B. bassiana isolate has a significant antagonistic potential in suppressing B. cinerea, which is the starting point for further research in vivo.

This work describes the characterization of Trichoderma atroviride strain CMU-08, isolated from Michoacán, Mexico. CMU-08 demonstrated robust growth and conidiation across a temperature range from 16 to 32 °C and a pH range from 4 to 9 on potato dextrose agar (PDA) and malt extract agar (MEA) media. The strain is an efficient antagonist of six species of phytopathogenic fungi and oomycetes in PDA, MEA, and Vogel minimal medium (VMM). Antagonist mechanisms of CMU-08 included direct mycoparasitism observed in dual-culture assays, as well as antibiosis attributed to growth inhibition via both volatile and non-volatile metabolites, with the effectiveness varying depending on the test phytopathogen and culture medium. Extracellular filtrates (ECFs) recovered from liquid cultures of CMU-08 under basal and induced conditions using Botrytis cinerea cell walls significantly inhibited their growth at a concentration of 750 µg/mL. Moreover, in detached tomato leaf assays, these ECFs reduced foliar damage caused by B. cinerea by 24–34%. The volatile organic compounds (VOCs) produced by CMU-08 also exhibited substantial efficacy, reducing foliar damage by up to 50% in similar tests. Despite showing no basal extracellular chitinase enzymatic activity, CMU-08 demonstrated significant induction of this activity in cultures supplemented with B. cinerea and Fusarium sp. cell walls. Four genes encoding extracellular chitinases (chit33, chit36, ech42, and locus 217415) showed different dynamics of transcriptional regulation during the dual-culture confrontation of strain CMU-08 with B. cinerea and Fusarium sp., varying according to the phytopathogen and the interaction stage. The CMU-08 strain shows physiological versatility and employs a variety of antagonist mechanisms toward different species of phytopathogenic microorganisms, making it a good candidate for developing a biocontrol product for field application. [ABSTRACT FROM AUTHOR]

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Black point disease of wheat contributes to high economic losses every year. The overuse of fungicides resulted in pathogenic fungal resistance, health and environmental hazards that necessitate the formulation of safe biocontrol agents. Dual cultural assay of Trichoderma viride and Trichoderma harzianum strains proved their highest efficiency against Alternaria alternata with inhibition percentages of 75.04 and 67.83%, while the lowest activity was detected against Drechslera halodes with growth inhibition of 51.54 and 43.92%, respectively. Alternaria alternata and D. halodes strains exhibited carbendazim resistance, while Fusarium proliferatum was the most susceptible one. Chemical analysis of T. harzianum and T. viride extracts using gas chromatography-mass spectrometry indicated that 6-pentyl-α-pyrone and cyclooctanol compounds were the main active constituents representing about 26.43 and 32.74% respectively. The tested bioagents were highly effective against the fungal strains causing black point disease of wheat so that these biocontrol agents could be used in formulation of natural fungicides avoiding the harmful impact of the synthetic fungicides. Abbreviations: FCZ; Fluconazole Antifungal agent, GC-MS; Gas chromatography-Mass spectrometry, MIC; Minimum inhibitory concentration, MFC; Minimum fungicidal concentration, PDA; Potato dextrose agar, T.H; Trichoderma harzianum, 1. T. V; Trichoderma viride

Abstract Background Avocado “tristeza,” a disease caused by Phytophthora cinnamomi, is one of the main limiting factors of avocado production in the Caribbean region. To control the pathogen, the application of agrochemicals is required, but this has caused environmental problems. Trichoderma spp. present properties in promoting plant growth and controlling phytopathogens, being proposed as an alternative to replace chemical fertilizers. The objective of the study was to evaluate the antagonistic activity of Trichoderma spp. against P. cinnamomi and its possible potential for promoting plant growth in vitro. Soil samples were taken from avocado cultivars from the municipalities of Ovejas (Sucre-Colombia) and Chalán (Sucre-Colombia). Results Serial dilutions were carried out for the isolation of Trichoderma spp. Once the strains were purified, the antagonism test against P. cinnamomi was carried out in PDA culture medium. For growth promotion, SRS medium was used for phosphate solubilization and CAS medium for siderophore production. DNA extraction and identification of the isolates were performed using the tef1 gene. Trichoderma harzianum and T. asperellum presented a 93.4% inhibition against the pathogen, followed by T. viride with an inhibition of 83.5% and finally T. longibrachiatum with 78.4% inhibition, showing significant differences in the control of the growth of the pathogen (p

Introduction: The fungal secretome comprise diverse proteins that are involved in various aspects of fungal lifestyles, including adaptation to ecological niches and environmental interactions. The aim of this study was to investigate the composition and activity of fungal secretomes in mycoparasitic and beneficial fungal-plant interactions. Methods: We used six Clonostachys spp. that exhibit saprotrophic, mycotrophic and plant endophytic lifestyles. Genome-wide analyses was performed to investigate the composition, diversity, evolution and gene expression of Clonostachys secretomes in relation to their potential role in mycoparasitic and endophytic lifestyles. Results and discussion: Our analyses showed that the predicted secretomes of the analyzed species comprised between 7 and 8% of the respective proteomes. Mining of transcriptome data collected during previous studies showed that 18% of the genes encoding predicted secreted proteins were upregulated during the interactions with the mycohosts Fusarium graminearum and Helminthosporium solani. Functional annotation of the predicted secretomes revealed that the most represented protease family was subclass S8A (11–14% of the total), which include members that are shown to be involved in the response to nematodes and mycohosts. Conversely, the most numerous lipases and carbohydrate-active enzyme (CAZyme) groups appeared to be potentially involved in eliciting defense responses in the plants. For example, analysis of gene family evolution identified nine CAZyme orthogroups evolving for gene gains (p ≤ 0.05), predicted to be involved in hemicellulose degradation, potentially producing plant defense-inducing oligomers. Moreover, 8–10% of the secretomes was composed of cysteine-enriched proteins, including hydrophobins, important for root colonization. Effectors were more numerous, comprising 35–37% of the secretomes, where certain members belonged to seven orthogroups evolving for gene gains and were induced during the C. rosea response to F. graminearum or H. solani. Furthermore, the considered Clonostachys spp. possessed high numbers of proteins containing Common in Fungal Extracellular Membranes (CFEM) modules, known for their role in fungal virulence. Overall, this study improves our understanding of Clonostachys spp. adaptation to diverse ecological niches and establishes a basis for future investigation aiming at sustainable biocontrol of plant diseases. [ABSTRACT FROM AUTHOR]

In the present work, research tasks were carried out in the search for fungi with potential biocontrol possibilities in relation to the ash dieback pathogen, Hymenoscyphus fraxineus. In the years 2012–2021, dead petioles of F. excelsior and F. mandshurica were collected, on which morphological structures of H. fraxineus showed unusual symptoms of dying (apothecia) and signs of colonization by other fungi (pseudosclerotial plates). Based on morphological and molecular phylogenetic data, 18 fungal taxa were identified. Thirteen of them belong to Ascomycota: Clonostachys rosea, Cl. solani, Cordyceps sp., Minimidochium sp., Nemania diffusa, Fusarium sp., Pestalotiopsis sp., Trichoderma atroviride, T. harzianum, T. polysporum, T. rodmanii, T. tomentosum, Trichoderma sp., and five other taxa are represented by Basidiomycota: Corticiales sp., Cyathus olla, Efibula sp., Gymnopus sp. and Polyporales sp. In 108 dual cultures in vitro, three different types of interactions were distinguished: (i) physical colony contact (5.6%), (ii) presence of an inhibition zone between the colonies (0.9%), and (iii) copartner overgrowth of H. fraxineus colonies and partial or complete replacement of the pathogen (93.5%). In the dual cultures, various morphological deformations of H. fraxineus hyphae were observed: the development of apical or intercalary cytoplasmic extrusions, development of internal hyphae of the test fungi in pathogens' hyphae, the deformation and disruption of significant sections of H. fraxineus hyphae via lysis and mycoparasitism, complete desolation of H. fraxineus cells and breakdown of hyphae into short fragments, and disappearing of pigment in the affected hyphae of H. fraxineus. The inoculation tests performed in vivo or in glass Petrie dishes showed that all the identified taxa were able to lead to pathological changes in H. fraxineus apothecia, and the mycelium of some of them completely covered pseudosclerotial plates of H. fraxineus. It was emphasized in the discussion that such activity of these fungi in forest stands may contribute to the reduction in the H. fraxineus inoculum reservoir. [ABSTRACT FROM AUTHOR]

A wide range of phytopathogenic fungi exist causing various plant diseases, which can lead to devastating economic, environmental, and social impacts on a global scale. One such fungus is Pyrrhoderma noxium, causing brown root rot disease in over 200 plant species of a variety of life forms mostly in the tropical and subtropical regions of the globe. The aim of this study was to discover the antagonistic abilities of two Trichoderma strains (#5001 and #5029) found to be closely related to Trichoderma reesei against P. noxium. The mycoparasitic mechanism of these Trichoderma strains against P. noxium involved coiling around the hyphae of the pathogen and producing appressorium like structures. Furthermore, a gene expression study identified an induced expression of the biological control activity associated genes in Trichoderma strains during the interaction with the pathogen. In addition, volatile and diffusible antifungal compounds produced by the Trichoderma strains were also effective in inhibiting the growth of the pathogen. The ability to produce Indole-3-acetic acid (IAA), siderophores and the volatile compounds related to plant growth promotion were also identified as added benefits to the performance of these Trichoderma strains as biological control agents. Overall, these results show promise for the possibility of using the Trichoderma strains as potential biological control agents to protect P. noxium infected trees as well as preventing new infections. [ABSTRACT FROM AUTHOR]

Trichoderma fungi have been proved as efficient bioagents with great antifungal properties while many species in the plant's rhizospheres have been characterized as plant growth-promoting agents. However, many rhizosphere Trichoderma are yet to be fully explored for plant disease management. In this study, Trichoderma species were isolated from the rhizosphere of maize, banana, and cassava, and their biocontrol potentials were screened against some Fusarium species from oak leaves (F2B and F3) and laboratory cultures (Fus 296 and Fus 294). The isolated rhizosphere Trichoderma were identified as Trichoderma virens 1 (TCIV), T. virens 2 (TCVII), T. virens 3 (TMSI), T. hazianum strain 1 (TCVI), T. harzianum strain 2 (TCVIII), T. erinaceum (TMZI), and T. koningiopsis (TMZII). The dual culture experiment recorded the highest percentage inhibition in TMZII against OakF2B (31.17%), TCVIII against Fus 294 (45.18%), TMZI against Fus 296 (47.37%), while TCIV was most effective against Oak F3 (44.15%). Among the Trichoderma culture filtrates evaluated, TCIV showed the highest percentage inhibition against Oak F3 (52.39%), Oak F2B (48.54%), Fus 294 (46.65%), and Fus 296 (44.48%). All the Trichoderma isolates demonstrated expressed varying levels of antagonism against the Fusarium pathogens in vitro. [ABSTRACT FROM AUTHOR]

Frosty pod rot, caused by the fungus Moniliophthora roreri, is one of the most important diseases of cacao in Peru. Sustainable alternatives to control it include timely cultural practices such as pruning, and the application of biological control agents such as Trichoderma spp. We isolated 234 Trichoderma strains native to the department of Amazonas in Northern Peru from soil samples in cacao farms. These strains belong to at least eighteen species within four phylogenetic clades in the genus (Harzianum, Longibrachiatum, Hamatum, and Brevicompactum clades). We aimed to assess the in vitro biocontrol potential of these strains against M. roreri. We evaluated their mycoparasitism, antibiosis, and potential antagonism to select candidate strains for efficient biocontrol of M. roreri. We found evidence (Kruskal–Wallis test, p < 0.005) that strains belonging to the Harzianum clade tend to have higher mycoparasitism, antibiosis, and potential antagonism levels than strains in the Longibrachiatum and Hamatum clades. Additionally, this study constitutes the first report for antagonistic behavior against M. roreri for T. parareesei, T. lentiforme, T. orientale, T. asperelloides, T. inhamatum, T. zelobreve, T. afarasin, T. ghanense, T. rifaii, and T. breve. These results will be foundational for further M. roreri biocontrol studies. [ABSTRACT FROM AUTHOR]

Trichoderma fungi have been proved as efficient bioagents with great antifungal properties while many species in the plant’s rhizospheres have been characterized as plant growth-promoting agents. However, many rhizosphere Trichoderma are yet to be fully explored for plant disease management. In this study, Trichoderma species were isolated from the rhizosphere of maize, banana, and cassava, and their biocontrol potentials were screened against some Fusarium species from oak leaves (F2B and F3) and laboratory cultures (Fus 296 and Fus 294). The isolated rhizosphere Trichoderma were identified as Trichoderma virens 1 (TCIV), T. virens 2 (TCVII), T. virens 3 (TMSI), T. hazianum strain 1 (TCVI), T. harzianum strain 2 (TCVIII), T. erinaceum (TMZI), and T. koningiopsis (TMZII). The dual culture experiment recorded the highest percentage inhibition in TMZII against OakF2B (31.17%), TCVIII against Fus 294 (45.18%), TMZI against Fus 296 (47.37%), while TCIV was most effective against Oak F3 (44.15%). Among the Trichoderma culture filtrates evaluated, TCIV showed the highest percentage inhibition against Oak F3 (52.39%), Oak F2B (48.54%), Fus 294 (46.65%), and Fus 296 (44.48%). All the Trichoderma isolates demonstrated expressed varying levels of antagonism against the Fusarium pathogens in vitro.

Apple valsa canker, caused by the fungus Valsa ceratosperma, is one of the most destructive diseases of this crop. Conventional fungicide treatments are not effective enough; therefore, it is necessary to develop alternatives for the control of this pathogen. The use of Trichoderma strains as a biocontrol agent is a promising strategy for the environmentally friendly management of this disease. In this study, seven different isolates of Trichoderma spp. were obtained from the rhizospheric soil of healthy apple plants. The antagonistic capacity of the isolates against the pathogen V. ceratosperma was evaluated using the dual culture method and by the production of antibiotic non-volatile compounds. In the dual culture method, the interaction zone was observed by differential interference contrast microscopy and confocal laser scanning microscopy. The isolates RN-13, RN-33, and RN-34 inhibited more than 75% of mycelial growth, whereas RN-19, RN-16, and RN-15 inhibited growth by 65–72% with respect to the control. The mycoparasitic activity was evidenced by the coiling of hyphae around pathogen hyphae, appressorium-like structures, morphological deformations, and disintegration of mycelial walls. Apple shoots inoculated with Trichoderma isolates and then infected with V. ceratosperma showed the antagonist capacity of RN-16, RN-19, RN-15, and RN-18, which produced a significant decrease in the size of the lesion. Overall, the results showed the great potential of Trichoderma as a biocontrol agent against apple tree valsa canker. Further studies of this fungal interaction are required to design effective control strategies for the protection of apple tree orchards from this disease. [ABSTRACT FROM AUTHOR]

Necrotrophic mycoparasitism is an intricate process involving recognition, physical mycelial contact, and killing of host fungi (mycohosts). During such interactions, mycoparasites undergo a complex developmental process involving massive regulatory changes of gene expression to produce a range of chemical compounds and proteins that contribute to the parasitism of the mycohosts. Small RNAs (sRNAs) are vital components of posttranscriptional gene regulation, although their role in gene expression regulation during mycoparasitisms remain understudied. Here, we investigated the role of sRNA-mediated gene regulation in mycoparasitism by performing sRNA and degradome tag sequencing of the mycoparasitic fungus Clonostachys rosea interacting with the plant-pathogenic mycohosts Botrytis cinerea and Fusarium graminearum at two time points. The majority of differentially expressed sRNAs were downregulated during the interactions with the mycohosts compared to a C. rosea self-interaction control, thus allowing desuppression (upregulation) of mycohost-responsive genes. Degradome analysis showed a positive correlation between high degradome counts and antisense sRNA mapping and led to the identification of 201 sRNA-mediated potential gene targets for 282 differentially expressed sRNAs. Analysis of sRNA potential gene targets revealed that the regulation of genes coding for membrane proteins was a common response against both mycohosts. The regulation of genes involved in oxidative stress tolerance and cellular metabolic and biosynthetic processes was exclusive against F. graminearum, highlighting common and mycohost-specific gene regulation of C. rosea. By combining these results with transcriptome data collected during a previous study, we expand the understanding of the role of sRNA in regulating interspecific fungal interactions and mycoparasitism. [ABSTRACT FROM AUTHOR]

Abstract Various strains of the mycoparasitic fungal species Clonostachys rosea are used commercially as biological control agents for the control of fungal plant diseases in agricultural crop production. Further improvements of the use and efficacy of C. rosea in biocontrol require a mechanistic understanding of the factors that determines the outcome of the interaction between C. rosea and plant pathogenic fungi. Here, we determined the genome sequences of 11 Clonostachys strains, representing five species in Clonostachys subgenus Bionectria, and performed a comparative genomic analysis with the aim to identify gene families evolving under selection for gene gains or losses. Several gene families predicted to encode proteins involved in biosynthesis of secondary metabolites, including polyketide synthases, nonribosomal peptide syntethases and cytochrome P450s, evolved under selection for gene gains (p ≤ .05) in the Bionectria subgenus lineage. This was accompanied with gene copy number increases (p ≤ .05) in ATP‐binding cassette (ABC) transporters and major facilitator superfamily (MFS) transporters predicted to contribute to drug efflux. Most Clonostachys species were also characterized by high numbers of auxiliary activity (AA) family 9 lytic polysaccharide monooxygenases, AA3 glucose–methanol–choline oxidoreductases and additional carbohydrate‐active enzyme gene families with putative activity (or binding) towards xylan and rhamnose/pectin substrates. Particular features of the C. rosea genome included expansions (p ≤ .05) of the ABC‐B4 multidrug resistance transporters, the ABC‐C5 multidrug resistance‐related transporters and the 2.A.1.3 drug:H + antiporter‐2 MFS drug resistance transporters. The ABC‐G1 pleiotropic drug resistance transporter gene abcG6 in C. rosea was induced (p ≤ .009) by exposure to the antifungal Fusarium mycotoxin zearalenone (1121‐fold) and various fungicides. Deletion of abcG6 resulted in mutants with reduced (p

Black point disease of wheat contributes to high economic losses every year. The overuse of fungicides resulted in pathogenic fungal resistance, health and environmental hazards that necessitate the formulation of safe biocontrol agents. Dual cultural assay of Trichoderma viride and Trichoderma harzianum strains proved their highest efficiency against Alternaria alternata with inhibition percentages of 75.04 and 67.83%, while the lowest activity was detected against Drechslera halodes with growth inhibition of 51.54 and 43.92%, respectively. Alternaria alternata and D. halodes strains exhibited carbendazim resistance, while Fusarium proliferatum was the most susceptible one. Chemical analysis of T. harzianum and T. viride extracts using gas chromatography-mass spectrometry indicated that 6-pentyl-α-pyrone and cyclooctanol compounds were the main active constituents representing about 26.43 and 32.74% respectively. The tested bioagents were highly effective against the fungal strains causing black point disease of wheat so that these biocontrol agents could be used in formulation of natural fungicides avoiding the harmful impact of the synthetic fungicides. Abbreviations: FCZ; Fluconazole Antifungal agent, GC-MS; Gas chromatography-Mass spectrometry, MIC; Minimum inhibitory concentration, MFC; Minimum fungicidal concentration, PDA; Potato dextrose agar, T.H; Trichoderma harzianum, 1. T. V; Trichoderma viride [ABSTRACT FROM AUTHOR]

ABSTRACT Dicer-like proteins (DCLs) play a vital role in RNA interference (RNAi), by cleaving RNA filament into small RNAs. Although DCL-mediated RNAi can regulate interspecific communication between pathogenic/mutualistic organisms and their hosts, its role in mycoparasitic interactions is yet to be investigated. In this study, we deleted dcl genes in the mycoparasitic fungus Clonostachys rosea and characterize the functions of DCL-dependent RNAi in mycoparasitism. Deletion of dcl2 resulted in a mutant with reduced secondary metabolite production, antagonism toward the plant-pathogenic fungus Botrytis cinerea, and reduced ability to control Fusarium foot rot disease on wheat, caused by Fusarium graminearum. Transcriptome sequencing of the in vitro interaction between the C. rosea Δdcl2 strain and B. cinerea or F. graminearum identified the downregulation of genes coding for transcription factors, membrane transporters, hydrolytic enzymes, and secondary metabolites biosynthesis enzymes putatively involved in antagonistic interactions, in comparison with the C. rosea wild-type interaction. A total of 61 putative novel microRNA-like RNAs (milRNAs) were identified in C. rosea, and 11 were downregulated in the Δdcl2 mutant. In addition to putative endogenous gene targets, these milRNAs were predicted to target B. cinerea and F. graminearum virulence factor genes, which showed an increased expression during interaction with the Δdcl2 mutant incapable of producing the targeting milRNAs. In summary, this study constitutes the first step in elucidating the role of RNAi in mycoparasitic interactions, with important implications for biological control of plant diseases, and poses the base for future studies focusing on the role of cross-species RNAi regulating mycoparasitic interactions. IMPORTANCE Small RNAs mediated RNA interference (RNAi) known to regulate several biological processes. Dicer-like endoribonucleases (DCLs) play a vital role in the RNAi pathway by generating sRNAs. In this study, we investigated a role of DCL-mediated RNAi in interference interactions between mycoparasitic fungus Clonostachys rosea and the two fungal pathogens Botrytis cinerea and Fusarium graminearum (here called mycohosts). We found that the dcl mutants were not able to produce 11 sRNAs predicted to finetune the regulatory network of genes known to be involved in production of hydrolytic enzymes, antifungal compounds, and membrane transporters needed for antagonistic action of C. rosea. We also found C. rosea sRNAs putatively targeting known virulence factors in the mycohosts, indicating RNAi-mediated cross-species communication. Our study expanded the understanding of underlying mechanisms of cross-species communication during interference interactions and poses a base for future works studying the role of DCL-based cross-species RNAi in fungal interactions.

The highly diverse genus Trichoderma has provided many formulations that are alternatives to the chemical pesticides in agriculture. The present study was undertaken to investigate the biocontrol potential of eight Trichoderma species, T. atrobrunneum, T. guizhouense, T. paratroviride, T. pyramidale, T. rufobrunneum, T. simmonsii, T. thermophilum and T. viridulum, against the phytopathogenic fungus Rhizoctonia solani. Trichoderma isolates were first evaluated in vitro by dual culture tests for their antagonism, mycoparasitic ability and antifungal activity against R. solani. Their growth promoting potential was further assessed in relation to phosphate solubilization, indole acetic acid and siderophore production. Five of the isolates were selected and evaluated for their abilities to prompt plant growth and to control R. solani infecting Vigna unguiculata (cowpea) seedlings in vivo. Two most effective isolates, T. guizhouense 9185 and T. simmonsii 8702, significantly (P

In the present work, research tasks were carried out in the search for fungi with potential biocontrol possibilities in relation to the ash dieback pathogen, Hymenoscyphus fraxineus. In the years 2012–2021, dead petioles of F. excelsior and F. mandshurica were collected, on which morphological structures of H. fraxineus showed unusual symptoms of dying (apothecia) and signs of colonization by other fungi (pseudosclerotial plates). Based on morphological and molecular phylogenetic data, 18 fungal taxa were identified. Thirteen of them belong to Ascomycota: Clonostachys rosea, Cl. solani, Cordyceps sp., Minimidochium sp., Nemania diffusa, Fusarium sp., Pestalotiopsis sp., Trichoderma atroviride, T. harzianum, T. polysporum, T. rodmanii, T. tomentosum, Trichoderma sp., and five other taxa are represented by Basidiomycota: Corticiales sp., Cyathus olla, Efibula sp., Gymnopus sp. and Polyporales sp. In 108 dual cultures in vitro, three different types of interactions were distinguished: (i) physical colony contact (5.6%), (ii) presence of an inhibition zone between the colonies (0.9%), and (iii) copartner overgrowth of H. fraxineus colonies and partial or complete replacement of the pathogen (93.5%). In the dual cultures, various morphological deformations of H. fraxineus hyphae were observed: the development of apical or intercalary cytoplasmic extrusions, development of internal hyphae of the test fungi in pathogens’ hyphae, the deformation and disruption of significant sections of H. fraxineus hyphae via lysis and mycoparasitism, complete desolation of H. fraxineus cells and breakdown of hyphae into short fragments, and disappearing of pigment in the affected hyphae of H. fraxineus. The inoculation tests performed in vivo or in glass Petrie dishes showed that all the identified taxa were able to lead to pathological changes in H. fraxineus apothecia, and the mycelium of some of them completely covered pseudosclerotial plates of H. fraxineus. It was emphasized in the discussion that such activity of these fungi in forest stands may contribute to the reduction in the H. fraxineus inoculum reservoir.

Background: High losses of sorghum crops due to fungal diseases, such as grain mold and stalk rot, are economically harmful and cause increased use of environmentally damaging chemical fungicides, which also are detrimental to human health. Hence, finding safe and effective ways to manage fungal diseases of sorghum is urgently needed. Results: In the present study, the antagonistic activities of Trichoderma viride and T. harzianum against different pathogenic fungal strains were evaluated in vitro using a dual culture assay. Furthermore, the anti-mycotic activity of Trichoderma spp. culture filtrates was evaluated against different fungal strains using a food poisoning technique. Additionally, the antifungal activities of ethyl acetate extracts of T. viride and T. harzianum against different pathogens were evaluated using a disk diffusion method. As indicated by the dual culture assay, T. harzianum suppressed 66.8, 69.5, 68.7, 54.6, 84.12, and 71.39% of the mycelial growth of Curvularia lunata, Exserohilum rostratum, Fusarium chlamydosporum, F. incarnatum, F. proliferatum, and Macrophomina phaseolina, respectively. T. viride was more effective for controlling the growth of these pathogens, inhibiting 81.0, 89.0, 63.0, 70.7, 84.4, and 71.8% of mycelial growth, respectively. Both E. rostratum and M. phaseolina showed resistance to carbendazim fungicide at all tested concentrations, whereas the fungicidal concentrations of carbendazim against C. lunata, F. chlamydosporum, and F. incarnatum strains were 2.50, 1.50, and 2.00 ppm, respectively. Furthermore, F. proliferatum was sensitive to carbendazim fungicide at all tested concentrations. Antifungal assays of the ethyl acetate extracts of T. viride and T. harzianum indicated the potent activity of these extracts against fungal phytopathogens with different susceptibility patterns. F. chlamydosporum was the most sensitive to the extracts of T. viride and T. harzianum with minimum inhibitory concentrations of 0.5 and 1.0 mg/disk, respectively. Conclusion: The potent suppression of sorghum phytopathogens by T. viride and T. harzianum makes them potential sources of safe and effective natural fungicides compared to carbendazim fungicide. [ABSTRACT FROM AUTHOR]

This study aimed to identify four isolates of Trichoderma sp. (Ufra.T06, Ufra.T09, Ufra.T12, and Ufra.T52) and characterize their interaction with Magnaporthe oryzae in vitro and in vivo conditions. The four isolates of Trichoderma sp. were sequenced, investigated as an antagonist against M. oryzae in five Petri plate assays, and as an inhibitor of conidial germination appressoria formation. Finally, were quantified the lytic activity of chitinase (CHI), glucanase (GLU), and protease (PRO) during co-cultivation of Trichoderma sp. and M. oryzae. In vivo, leaf blast suppression was evaluated in two assays: simultaneous and curative application. Both in vitro and in vivo assays were scanned by electron microscopy (SEM). All isolates were identified as Trichoderma asperellum. All in vitro Petri plates assays reduced M. oryzae colony growth (paired–91.18% by Ufra.T09, volatile metabolites—all isolates equally reduced, non-volatile—68.33% by Ufra.T06, thermostability—99.77% by Ufra.T52 and co-cultivate—64.25% by Ufra.T52). The filtrates and conidia suspensions for T. asperellum isolates inhibited the conidia germination and appressoria formation significantly. In co-cultivate (mycelial or cell wall), all enzymes (GLU, CHI, and PRO) and times (24, 48, and 72 h) showed increased activity. In vivo, reduced leaf blast severity until 94.64% (Ufra.T52cs) in a simultaneous and until 85% (Ufra.T09 24 and 48 hasi) in a curative application. T. asperellum isolates showed efficient control of M. oryzae by mycoparasitism, and antibiosis mechanisms were interfered with by the M. oryzae infection process. [ABSTRACT FROM AUTHOR]

Various strains of the mycoparasitic fungal species Clonostachys rosea are used commercially as biological control agents for the control of fungal plant diseases in agricultural crop production. Further improvements of the use and efficacy of C. rosea in biocontrol require a mechanistic understanding of the factors that determines the outcome of the interaction between C. rosea and plant pathogenic fungi. Here, we determined the genome sequences of 11 Clonostachys strains, representing five species in Clonostachys subgenus Bionectria, and performed a comparative genomic analysis with the aim to identify gene families evolving under selection for gene gains or losses. Several gene families predicted to encode proteins involved in biosynthesis of secondary metabolites, including polyketide synthases, nonribosomal peptide syntethases and cytochrome P450s, evolved under selection for gene gains (p ≤.05) in the Bionectria subgenus lineage. This was accompanied with gene copy number increases (p ≤.05) in ATP‐binding cassette (ABC) transporters and major facilitator superfamily (MFS) transporters predicted to contribute to drug efflux. Most Clonostachys species were also characterized by high numbers of auxiliary activity (AA) family 9 lytic polysaccharide monooxygenases, AA3 glucose–methanol–choline oxidoreductases and additional carbohydrate‐active enzyme gene families with putative activity (or binding) towards xylan and rhamnose/pectin substrates. Particular features of the C. rosea genome included expansions (p ≤.05) of the ABC‐B4 multidrug resistance transporters, the ABC‐C5 multidrug resistance‐related transporters and the 2.A.1.3 drug:H + antiporter‐2 MFS drug resistance transporters. The ABC‐G1 pleiotropic drug resistance transporter gene abcG6 in C. rosea was induced (p ≤.009) by exposure to the antifungal Fusarium mycotoxin zearalenone (1121‐fold) and various fungicides. Deletion of abcG6 resulted in mutants with reduced (p <.001) growth rates on media containing the fungicides boscalid, fenhexamid and iprodione. Our results emphasize the role of biosynthesis of, and protection against, secondary metabolites in Clonostachys subgenus Bionectria. [ABSTRACT FROM AUTHOR]

Aegerolysins are small secreted pore-forming proteins that are found in both prokaryotes and eukaryotes. The role of aegerolysins in sporulation, fruit body formation, and in lysis of cellular membrane is suggested in fungi. The aim of the present study was to characterize the biological function of the aegerolysin gene agl1 in the mycoparasitic fungus Trichoderma atroviride, used for biological control of plant diseases. Gene expression analysis showed higher expression of agl1 during conidiation and during growth in medium supplemented with cell wall material from the plant pathogenic fungus Rhizoctonia solani as the sole carbon source. Expression of agl1 was supressed under iron-limiting condition, while agl1 transcript was not detected during T. atroviride interactions with the prey fungi Botrytis cinerea or R. solani. Phenotypic analysis of agl1 deletion strains (Δagl1) showed reduced conidiation compared to T. atroviride wild type, thus suggesting the involvement of AGL1 in conidiation. Furthermore, the Δagl1 strains display reduced antagonism towards B. cinerea and R. solani based on a secretion assay, although no difference was detected during direct interactions. These data demonstrate the role of AGL1 in conidiation and antagonism in the mycoparasitic fungus T. atroviride. [ABSTRACT FROM AUTHOR]

Lysin motif (LysM) modules are approximately 50 amino acids long and bind to peptidoglycan, chitin and its derivatives. Certain LysM proteins in plant pathogenic and entomopathogenic fungi are shown to scavenge chitin oligosaccharides and thereby dampen host defense reactions. Other LysM proteins can protect the fungal cell wall against hydrolytic enzymes. In this study, we investigated the biological function of LysM proteins in the mycoparasitic fungus Clonostachys rosea. The C. rosea genome contained three genes coding for LysM-containing proteins and gene expression analysis revealed that lysm1 and lysm2 were induced during mycoparasitic interaction with Fusarium graminearum and during colonization of wheat roots. Lysm1 was suppressed in germinating conidia, while lysm2 was induced during growth in chitin or peptidoglycan-containing medium. Deletion of lysm1 and lysm2 resulted in mutants with increased levels of conidiation and conidial germination, but reduced ability to control plant diseases caused by F. graminearum and Botrytis cinerea. The Δ lysm2 strain showed a distinct, accelerated mycelial disintegration phenotype accompanied by reduced biomass production and hyphal protection against hydrolytic enzymes including chitinases, suggesting a role of LYSM2 in hyphal protection against chitinases. The Δ lysm2 and Δ lysm1 Δ lysm2 strains displayed reduced ability to colonize wheat roots, while only Δ lysm1 Δ lysm2 failed to suppress expression of the wheat defense response genes PR1 and PR4. Based on our data, we propose a role of LYSM1 as a regulator of fungal development and of LYSM2 in cell wall protection against endogenous hydrolytic enzymes, while both are required to suppress plant defense responses. Our findings expand the understanding of the role of LysM proteins in fungal-fungal interactions and biocontrol. [ABSTRACT FROM AUTHOR]

The plant-Trichoderma-pathogen triangle is a complicated web of numerous processes. Trichoderma spp. are avirulent opportunistic plant symbionts. In addition to being successful plant symbiotic organisms, Trichoderma spp. also behave as a low cost, effective and ecofriendly biocontrol agent. They can set themselves up in various patho-systems, have minimal impact on the soil equilibrium and do not impair useful organisms that contribute to the control of pathogens. This symbiotic association in plants leads to the acquisition of plant resistance to pathogens, improves developmental processes and yields and promotes absorption of nutrient and fertilizer use efficiency. Among other biocontrol mechanisms, antibiosis, competition and mycoparasitism are among the main features through which microorganisms, including Thrichoderma, react to the presence of other competitive pathogenic organisms, thereby preventing or obstructing their development. Stimulation of every process involves the biosynthesis of targeted metabolites like plant growth regulators, enzymes, siderophores, antibiotics, etc. This review summarizes the biological control activity exerted by Trichoderma spp. and sheds light on the recent progress in pinpointing the ecological significance of Trichoderma at the biochemical and molecular level in the rhizosphere as well as the benefits of symbiosis to the plant host in terms of physiological and biochemical mechanisms. From an applicative point of view, the evidence provided herein strongly supports the possibility to use Trichoderma as a safe, ecofriendly and effective biocontrol agent for different crop species.

Abstract Protoplast fusion is an imperative tool to develop Trichoderma inter-fusants having desire traits through genetic manipulation. Study designed to develop diverse Trichoderma fusants for fungicide tolerance (Mancozeb, Thiram, Tebuconazole, and Carbendazim) and enhanced mycoparasitic activity against Sclerotium rolfsii sacc. The mycoparasitic T. virens NBAII Tvs12 and fungicide tolerant T. koningii MTCC 796 were utilized for protoplast fusion. The derived inter-fusants were subjected to diploidization using d-camphor in minimal media followed by successive three sub culturing onto potato dextrose agar to obtain 36 stable fusants. The stable fusants were employed for conidial size, fungicide tolerance, mycoparasitism, gene specific SSR amplification and molecular heterozygosity analysis. The results explained that 22 homozygous mutants illustrated characteristic of either one parental strain and 14 heterozygous recombinants depicted traits of both parental strains. The antagonistic activity of fusants against S. rolfsii depicted highest growth inhibition (87.91%) by potent inter-fusant (Fu 21) with improved fungicide tolerance capacity. The molecular study revealed highest observed heterozygocity (0.544), coefficient of gene differentiation (0.526) and gene flow (0.387) by Fu 21 indicating better genetic exploitation of parental strains into that fusant with good genetic purity. Principal coordinate analysis of fusants and parental strains exhibited 65.07% total variation and confirmed the scattering pattern matched with UPGMA clustering pattern. The stable heterozygous Fu 21 derived from inter-fusion between Tvs 12 and MTCC 796 might be useful to practice eco-friendly bioformulation tolerance to fungicides for effective integrated stem rot disease management in groundnut. Graphical abstract Unlabelled Image Highlights • Novel stable heterozygous Fu 21 derived from inter-fusion of Trichoderma (T. virens NBAII Tvs 12 and T. koningii MTCC 796) • Molecular heterozygosity analysis of 36 fusants performed to confirm genetic status of fusants using gene specific SSR • The highest observed heterozygocity, coefficient of gene differentiation and gene flow observed in potent Fu 21 • The Fu 21 exhibited highest growth inhibition of S. rolfsii with enhanced fungicides tolerance capacity. • The Fu 21 might be useful to practice eco-friendly bioformulation tolerance to fungicides for disease management. [ABSTRACT FROM AUTHOR]

Background: So far, biocontrol agent selection has been performed mainly by time consuming in vitro confrontation tests followed by extensive trials in greenhouse and field. An alternative approach is offered by application of high-throughput techniques, which allow extensive screening and comparison among strains for desired genetic traits. In the genus Trichoderma, the past assignments of particular features or strains to one species need to be reconsidered according to the recent taxonomic revisions. Here we present the genome of a biocontrol strain formerly known as Trichoderma harzianum ITEM 908, which exhibits both growth promoting capabilities and antagonism against different fungal pathogens, including Fusarium graminearum, Rhizoctonia solani, and the root-knot nematode Meloidogyne incognita. By genomic analysis of ITEM 908 we investigated the occurrence and the relevance of genes associated to biocontrol and stress tolerance, providing a basis for future investigation aiming to unravel the complex relationships between genomic endowment and exhibited activities of this strain. Results: The MLST analysis of ITS-TEF1 concatenated datasets reclassified ITEM 908 as T. atrobrunneum, a species recently described within the T. harzianum species complex and phylogenetically close to T. afroharzianum and T. guizhouense. Genomic analysis revealed the presence of a broad range of genes encoding for carbohydrate active enzymes (CAZYmes), proteins involved in secondary metabolites production, peptaboils, epidithiodioxopiperazines and siderophores potentially involved in parasitism, saprophytic degradation as well as in biocontrol and antagonistic activities. This abundance is comparable to other Trichoderma spp. in the T. harzianum species complex, but broader than in other biocontrol species and in the species T. reesei, known for its industrial application in cellulase production. Comparative analysis also demonstrated similar genomic organization of major secondary metabolites clusters, as in other Trichoderma species. Conclusions: Reported data provide a contribution to a deeper understanding of the mode of action and identification of activity-specific genetic markers useful for selection and improvement of biocontrol strains. This work will also enlarge the availability of genomic data to perform comparative studies with the aim to correlate phenotypic differences with genetic diversity of Trichoderma species. [ABSTRACT FROM AUTHOR]

Leaf-cutter ants cultivate and feed on the mutualistic fungus, Leucoagaricus gongylophorus, which is threatened by parasitic fungi of the genus Escovopsis. The mechanism of Escovopsis parasitism is poorly understood. Here, we assessed the nature of the antagonism of different Escovopsis species against its host. We also evaluated the potential antagonism of Escovopsioides, a recently described fungal genus from the attine ant environment whose role in the colonies of these insects is unknown. We performed dual-culture assays to assess the interactions between L. gongylophorus and both fungi. We also evaluated the antifungal activity of compounds secreted by the latter on L. gongylophorus growth using crude extracts of Escovopsis spp. and Escovopsioides nivea obtained either in (1) absence or (2) presence of the mutualistic fungus. The physical interaction between these fungi and the mutualistic fungus was examined under scanning electron microscopy (SEM). Escovopsis spp. and E. nivea negatively affected the growth of L. gongylophorus, which was also significantly inhibited by both types of crude extract. These results indicate that Escovopsis spp. and E. nivea produce antifungal metabolites against the mutualistic fungus. SEM showed that Escovopsis spp. and E. nivea maintained physical contact with the mutualistic fungus, though no specialised structures related to mycoparasitism were observed. These results showed that Escovopsis is a destructive mycoparasite that needs physical contact for the death of the mutualistic fungus to occur. Also, our findings suggest that E. nivea is an antagonist of the ant fungal cultivar. [ABSTRACT FROM AUTHOR]

Cultivos de maracuyá de la región Caribe colombiana presentan problemas de marchitez y el hongo Fusarium solani (Mart.) Sacc., está asociado a esta patología. Plantas de maracuyá de la Estación Experimental (E.E Caribia de Corpoica) poseen este problema y por tal razón se determinó el antagonismo in vitro de aislamientos de Trichoderma harzianum Rifai, frente a F. solani, aislado de plantas enfermas de maracuyá (Passiflora edulis). Utilizando la técnica de cultivo dual en platos Petri con Agar Sabouraud, se evaluaron competencia por nutrientes y espacio, micoparasitismo y porcentaje de inhibición del crecimiento radial (PICR), empleando un diseño estadístico factorial 2x6x1 con arreglo completamente aleatorio. Se obtuvieron tres aislamientos nativos de T. harzianum (TCN-009, TCN-010, TCN-014) de suelo de Palma de Aceite (Elaeis guineensis) de la E.E Caribia, y se compararon con tres aislamientos comerciales (TCC-001, TCC-005, TCC-006). TCC-001 y TCN-014, reportaron ser más competentes por nutrientes y espacio, con el mayor radio de crecimiento de 7,50 y 7,32 cm el día 10, comparado a FSM-011 en el cual solo fue de 2,30 cm. Aunque, TCN-014 mostró micoparasitismo grado 4 con ambos aislamientos de F. solani y TCC-005 únicamente con FSM-012, el cual fue más susceptible a ser micoparasitado. En cuanto al PICR, los tratamientos con mejores porcentajes de inhibición fueron TA-9, TA-12 y TA-6 con valores de 70,56, 68,52 y 65,32% respectivamente. El aislamiento del patógeno mayormente inhibido fue FSM-011. Todos estos resultados demuestran que hubo antagonismo in vitro al utilizar los aislamientos nativos y comerciales de T. harzianum sobre F. solani. Palabras clave: Passiflora, competencia, micoparasitismo, inhibición, antagonismo.

Trichoderma spp. is a fungus of the Ascomycota division with great potential to control diseases in economically important crops. This antagonistic fungus inhibits the growth of phytopathogenic microorganisms, including those responsible for root diseases, such as Rhizoctonia spp. and Fusarium spp. The objective of this study was to identify Trichoderma species and to describe their in vitro antagonistic activity in pathogenic strains of Rhizoctonia solani, F. oxysporum, F. verticillioides and F. solani, with different virulence determined based on pathogenicity tests in bean plants (Phaseolus vulgaris L.) cv. Montcalm. The hypothesis was that the identification and antagonistic description of Trichoderma species against pathogenic fungi defines their biological control efficiency. The experimental design was completely random, with at least three repetitions. The molecular identification of Trichoderma spp. was done through ITS1 and ITS2 regions, separated by the 5.8S ribosomal RNA gene (ITS, Internal Transcribed Spacer). The antagonism was evaluated using the modified cellophane method and dual cultures to estimate the percentage of radial growth inhibition (PRGI). Seven Trichoderma strains identified using molecular techniques showed 95-100 % identity to T. harzianum and T. asperellum. Trichoderma strains showed high antagonistic capacity with 73.1 to 76.4 mean PRGI values in pathogenic strains, suggesting an antibiosis effect (77.6 PRGI) and different hyphal interaction types. Antagonistic strains were statistically similar for pathogen growth and development inhibition; however, T. asperellum's antagonistic values were higher than T. harzianum's with regard to all pathogenic strains, even R. solani, which was the most aggressive pathogen for cv. Montcalm (mean value: 7.0). The antagonistic effect of Trichoderma strains on phytopathogenic fungi was determined with in vitro tests and corroborated with in vivo bioassays with similar inhibition percentages. [ABSTRACT FROM AUTHOR]

Trichoderma spp. are used for biocontrol of several plant pathogens. However, their efficient interaction with the host needs to be accompanied by production of secondary metabolites and cell wall-degrading enzymes. Three parameters were evaluated after interaction between four Trichoderma species and plant-pathogenic fungi: Fusarium solani, Rhizoctonia solani and Sclerotinia sclerotiorum. Trichoderma harzianum and T. asperellum were the most effective antagonists against the pathogens. Most of the Trichoderma species produced toxic volatile metabolites, having significant effects on growth and development of the plant pathogens. When these species were grown in liquid cultures with cell walls from these plant pathogens, they produced and secreted β-1,3-glucanase, NAGAse, chitinase, acid phosphatase, acid proteases and alginate lyase. [ABSTRACT FROM AUTHOR]

Abstract: Some species of Trichoderma have successfully been used in the commercial biological control of fungal pathogens, e.g., Sclerotinia sclerotiorum, an economically important pathogen of common beans (Phaseolus vulgaris L.). The objectives of the present study were (1) to provide molecular characterization of Trichoderma strains isolated from the Brazilian Cerrado; (2) to assess the metabolic profile of each strain by means of Biolog FF Microplates; and (3) to evaluate the ability of each strain to antagonize S. sclerotiorum via the production of cell wall-degrading enzymes (CWDEs), volatile antibiotics, and dual-culture tests. Among 21 isolates, we identified 42.86 % as Trichoderma asperellum, 33.33 % as Trichoderma harzianum, 14.29 % as Trichoderma tomentosum, 4.76 % as Trichoderma koningiopsis, and 4.76 % as Trichoderma erinaceum. Trichoderma asperellum showed the highest CWDE activity. However, no species secreted a specific group of CWDEs. Trichoderma asperellum 364/01, T. asperellum 483/02, and T. asperellum 356/02 exhibited high and medium specific activities for key enzymes in the mycoparasitic process, but a low capacity for antagonism. We observed no significant correlation between CWDE and antagonism, or between metabolic profile and antagonism. The diversity of Trichoderma species, and in particular of T. harzianum, was clearly reflected in their metabolic profiles. Our findings indicate that the selection of Trichoderma candidates for biological control should be based primarily on the environmental fitness of competitive isolates and the target pathogen. [Copyright &y& Elsevier]

To establish the modes of action of the antagonistic fungal strain Clonostachys rosea BAFC3874 isolated from suppressive soils against Sclerotinia sclerotiorum and to determine its potential as a biocontrol agent. The antagonistic activity of C. rosea BAFC3874 was determined in vitro by dual cultures. The strain effectively antagonized S. sclerotiorum in pot-grown lettuce and soybean plants. Antifungal activity assays of C. rosea BAFC3874 grown in culture established that the strain produced antifungal compounds against S. sclerotiorum associated with secondary metabolism. High mycelial growth inhibition coincided with sclerotia production inhibition. The C. rosea strain produced a microheterogeneous mixture of peptides belonging to the peptaibiotic family. Moreover, mycoparasitism activity was observed in the dual culture. Clonostachys rosea strain BAFC3874 was proved to be an effective antagonist against the aggressive soil-borne pathogen S. sclerotiorum in greenhouse experiments. The main mechanisms involve peptaibiotic metabolite production and mycoparasitism activity. Clonostachys rosea BAFC3874 may be a good fungal biological control agent against S. sclerotiorum. In addition, we were also able to isolate and identify peptaibols, an unusual family of compounds in this genus of fungi. [ABSTRACT FROM AUTHOR]

Antagonist fungi play an important role in the biological control of phytopathogens. In this sense, Trichoderma genus is the most used for the biocontrol of soil fungus pathogens. These species present different action ways or mechanisms to carry out the control of phytopathogens. Among them are: competition for substrate, mycoparasitism, antibiosis, desactivation of enzymes of the pathogen, induced resistance, among others. While higher is the probability of a Trichoderma isolate to show several action ways, more efficient and lasting the control of the pathogen will be, aspects not being present in chemical pesticides. In the article, authors intend to carry out a compilation on the ways of action of Trichoderma against pathogens. [ABSTRACT FROM AUTHOR]

Pythium lycopersicum sp. nov. has been isolated from soil samples taken in an agricultural land in the Isparta region of Southern Turkey. This oomycete is characterized by its contiguous sporangia having globose to elongated elements linked with hyphal filaments, ornamented oogonia, and monoclinous antheridia with large antheridial cells. The oomycete is reminiscent of Pythium ornamentatum described by the corresponding author in 1987 from soil samples taken in Algeria. Sequence analyses of the internal transcribed spacer (ITS) regions of rRNA show a close relationship with Pythium oligandrum and other mycoparasites possessing ornamented oogonia. Morphological and molecular features of this isolate justify its description as a new species: P. lycopersicum. When grown together with Botrytis cinerea, this oomycete shows a pronounced antagonism and suppresses the phytopathogen. The morphological details, together with the analysis of the ITS region of its rRNA, and its antagonism with some phytopathogens are discussed in this article. [ABSTRACT FROM AUTHOR]

Studies were conducted to determine the effects of soil moisture (9, 16 or 24% w/w) and temperature (5, 15, 20 or 25°C) on the control of sclerotia of Sclerotinia sclerotiorum by five fungal agents in sterile and natural field soil. All five biocontrol agents were effective in reducing the survival of sclerotia of S. sclerotiorum in sterile soil under dry (9% moisture) or wet (24% moisture) conditions at 20°C, but only Coniothyrium minitans was effective in natural soil. Coniothyrium minitans was the most effective in reducing sclerotial viability at the temperature range of 15–25°C. Trichoderma virens was effective against sclerotia of S. sclerotiorum to a lesser extent than C. minitans, and in non-autoclaved soil, it performed best at 25°C. Although Epicoccum purpurascens, Talaromyces flavus and Trichothecium roseum were effective against sclerotia of S. sclerotiorum in some instances, they were less effective than C. minitans and T. virens. Sclerotia of S. sclerotiorum conditioned for myceliogenic germination were more vulnerable to attack by the biocontrol agents than dormant sclerotia. The implications are discussed with respect to enhancement of biological control of crop diseases caused by S. sclerotiorum in different geographic regions. [ABSTRACT FROM AUTHOR]

Different interactions between soil fungi competing in the rhizosphere with each other are necessary to understand their influence on plant growth and health. The interactions between the ectomycorrhizal (ECM) fungus Laccaria laccata and soil saprotrophic fungi (T. harzianum, T. virens) were studied by transmission electron microscopy, and by gold cytochemistry to assess the potential role of cell wall lytic enzymes in mycoparasitism. Anti-β-1,3-glucan antibody, WGA/ovomucoid-gold complex and PATAg test were used to localize β-1,3-glucan, chitin and polysaccharides. Cytoplasm disorganisation of the saprotrophic fungi occurred concurrently with dissolution of β-1,3-glucan in walls of hyphae and conidia of the saprotrophic fungi. Then digestion of polysaccharides and chitin of colonised fungal structures occurred. The studies suggest sequential contribution of cell wall lytic enzymes and importance of disturbing the host's cell integrity during mycoparasitism. We conclude that the ECM fungus can parasitise on the saprotrophic fungi not only in dual culture on artificial medium but also in the rhizosphere of Scots pine. [ABSTRACT FROM AUTHOR]

Abstract: Trichoderma is widely used as biocontrol agent against phytopathogenic fungi, and as biofertilizer because of its ability to establish mycorriza-like association with plants. The key factor to the ecological success of this genus is the combination of very active mycoparasitic mechanisms plus effective defense strategies induced in plants. This work, different from most of the studies carried out that address the attacking mechanisms, focuses on elucidating how Trichoderma is able to tolerate hostile conditions. A gene from Trichoderma harzianum CECT 2413, qid74, was strongly expressed during starvation of carbon or nitrogen sources; it encoded a cell wall protein of 74kDa that plays a significant role in mycelium protection. qid74 was originally isolated and characterized, in a previous work, by a differential hybridization approach under simulated mycoparasitism conditions. Heterologous expression of Qid74 in Saccharomyces cerevisiae indicated that the protein, located in the cell wall, interfered with mating and sporulation but not with cell integrity. The qid74 gene was disrupted by homologous recombination and it was overexpressed by isolating transformants selected for the amdS gene that carried several copies of qid74 gene under the control of the pki promoter. Disruptants and transformants showed similar growth rate and viability when they were cultivated in different media, temperatures and osmolarities, or were subjected to different abiotic stress conditions. However, disruptants produced about 70% mass yield under any condition and were substantially more sensitive than the wild type to cell wall degradation by different lytic preparations. Transformants had similar mass yield and were more resistant to lytic enzymes but more sensitive to copper sulfate than the wild type. When experiments of adherence to hydrophobic surfaces were carried out, the disruptants had a reduced capacity to adhere, whereas that capacity in the overproducer transformants was slightly higher than that of the wild type. Results point to a significant role for Qid74 both in cell wall protection and adhesion to hydrophobic surfaces. [Copyright &y& Elsevier]

Abstract: Frosty pod rot (FPR), caused by Moniliophthora roreri, and black pod (BP), caused by Phytophthora spp., of cocoa (Theobroma cacao) cause combined pod losses of more than 80% in Panama. Biological control of both diseases appeared promising in Peru and is desired by certified organic producers in Panama. We evaluated both local and Peruvian fungal antagonists in participatory trials on smallholdings during two complete production cycles. Furthermore, we tested the influence of a 3% v/v cane molasses amendment on biocontrol efficacy, yield and population dynamics of mycoparasites on the cocoa pod. Significant variation was observed between the two years: FPR was more severe in the first year, BP in the second. FPR was significantly reduced by biocontrol agents (BCAs), but not by the molasses amendment. However, BCAs responded differently to molasses in both years. All BCAs reduced inoculum production by M. roreri with no consistent effect of molasses. BCAs had a lesser and more variable effect on BP, whereas molasses reduced BP slightly by increasing the efficacy of native antagonists. All BCAs and the molasses amendment enhanced the percentage of healthy pods. Molasses was beneficial to absolute yield, but only one inoculum improved yield significantly in the first year. Populations of a Peruvian Trichoderma asperellum isolate remained high for over two months after application to surface-sterilized pods. Molasses had no effect on establishment or survival of this antagonist or recolonization by any native mycoparasite. The reasons for enhanced biocontrol efficacy of the molasses formulation requires further investigation. [Copyright &y& Elsevier]

Biological control agents for plant diseases are currently being examined as alternatives to synthetic pesticides due to their perceived increased level of safety and minimal environmental impacts. Fungal biological control agents have several mechanisms of action that allow them to control pathogens, including mycoparasitism, production of antibiotics or enzymes, competition for nutrients and the induction of plant host defences. While effective in the control of plant diseases, these mechanisms may pose risks to non-target species including mycorrhizal and saprophytic fungi, soil bacteria, plants, insects, aquatic and terrestrial animals, and humans. Non-target effects including mycoparasitism of mycorrhizae, reduction in plant root colonisation by mycorrhizal fungi, disorders in commercial mushrooms and nodulation by Rhizobium spp., and changes in plant growth have been associated with fungal biological control agents, such as Trichoderma spp. Also, the genera Trichoderma and Gliocladium have been linked to respiratory disorders and shellfish toxicity in humans, respectively. Biological control agents, such as Pythium oligandrum, Talaromyces flavus, Coniothyrium minitans and Ampelomyces quisqualis have modes of action which may pose risks to non-target fungi, bacteria, plants and animals. There is need for future research into ecological impacts associated with the release of any biological agent and methods of determining possible non-target effects. Adequate monitoring and the use of molecular techniques to identify and follow the movement of biological control agents are needed to examine and mitigate negative biological impacts. [Copyright &y& Elsevier]

Rosellinia spp. cause substantial mortality of cocoa (Theobroma cacao), especially in acidic soils rich in organic matter (O.M.) and, reportedly, low available phosphate. Cultural and chemical control are relatively ineffective and a supplementary biocontrol strategy would be highly desirable. Our aim was the isolation and evaluation of promising mycoparasites against Rosellinia of cocoa. We isolated 45 mycoparasites using a baiting technique. Three of them and a commercial isolate were prioritized for further evaluation based on their broad hostrange and/or effectiveness in pot bioassays. Because Rosellinia is favored by the same soil factors as mycoparasites, we investigated the effect of soil pH (range 4–6), O.M. (6–12%), and phosphate (0–1000 mg kg−1) on their interaction. In vitro, both pathogens and antagonists grew well on a wide range of phosphate (0–9500 mg kg−1) and at various pH (3.0–10.0) levels. The pH optimum usually coincided with the soil of origin and the phosphate optimum depended on the phosphate source used. In pot bioassays using natural soils, mixtures of antagonists exhibited more control than single strains and mixtures were effective over a wider range of soil conditions. High O.M. favored the pathogen more than its biocontrol agents. Soil pH interacted with the treatments such that biocontrol was most efficient at higher pH values and with the highest mixtures of antagonists. The addition of phosphate enhanced plant growth but had little effect on biocontrol efficacy. We conclude that mixtures of mycoparasites should be field tested in combination with liming and removal of O.M. [Copyright &y& Elsevier]