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The present study investigates the species boundaries, evolutionary relationships, and host-parasite interfaces of dimorphic mycoparasites that were previously assigned to Achroomyces soranus, Occultifur internus, and Platygloea mycophila based on morphological similarities. Our comparison of recently collected and cultivated samples with the type specimens of A. soranus and P. mycophila shows that both groups are species complexes, of which the taxa can be differentiated based on morphological and ecological characters. By integrating the results of a seven-locus dataset (SSU, LSU, ITS, RPB1, RPB2, TEF1, and mitochondrial CYT-B) and detailed micromorphological comparisons of the investigated specimens, we show for the first time that these three groups of mycoparasites belong to Cystobasidiomycetes (Pucciniomycotina). We applied a polyphasic species concept involving morphology, phylogeny, and ecology to delineate and circumscribe these and new genera. The genus Occultifur comprises six species. Occultifur internus and the newly proposed O. cerinomycicola are intrahymenial mycoparasites producing haustorial cells and establishing fusion pore interaction with their Dacrymycetous host. Based on microscopical examination, we show that Achroomyces soranus is a member of the genus Occultifur. Based on the molecular phylogenetic reconstruction, we found that three lichen-associated fungi which are only known from a yeast morph are nested within Occultifur, i.e. Lichenozyma pisutiana, Microsporomyces cladoniae, and M. wangii. The genus Obvidator is newly introduced for three mycoparasitic species inhabiting members of the corticioid genus Peniophora (Russulales, Agaricomycetes) and causing gall-like malformations of the host basidiome. Microscopic investigation shows that Platygloea mycophila is a member of this genus. Obvidator species display a yet undiscovered type of host-parasite interface, in which the mycoparasites produce short protrusions on their hyphae adjacent to the host hyphae. The lysis of the host cell wall takes place at points of contact with parasite protrusions, but no rupture of the host plasma membrane or nanometer-fusion pore formation was observed. The updated Cystobasidiomycetes phylogeny obtained in this study by including mycoparasites showed that the genera Occultifur and Microsporomyces as currently circumscribed are polyphyletic. To resolve this polyphyly, we introduce two new genera, i.e. Cystastrum and Millanizyma, and recombine species comprising the Occultifur externus clade and a clade consisting of Microsporomyces bloemfonteinensis and M. cladoniophilus, respectively. [ABSTRACT FROM AUTHOR]

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.

Abstract Background The application of plant-beneficial microorganisms as bio-fertilizer and biocontrol agents has gained traction in recent years, as both agriculture and forestry are facing the challenges of poor soils and climate change. Trichoderma spp. are gaining popularity in agriculture and forestry due to their multifaceted roles in promoting plant growth through e.g. nutrient translocation, hormone production, induction of plant systemic resistance, but also direct antagonism of other fungi. However, the mycotrophic nature of the genus bears the risk of possible interference with other native plant-beneficial fungi, such as ectomycorrhiza, in the rhizosphere. Such interference could yield unpredictable consequences for the host plants of these ecosystems. So far, it remains unclear, whether Trichoderma is able to differentiate between plant-beneficial and plant-pathogenic fungi during the process of plant colonization. Results We investigated whether Trichoderma spp. can differentiate between beneficial ectomycorrhizal fungi (represented by Laccaria bicolor and Hebeloma cylindrosporum) and pathogenic fungi (represented by Fusarium graminearum and Alternaria alternata) in different confrontation scenarios, including a newly developed olfactometer “race tube”-like system. Using two independent species, T. harzianum and T. atrobrunneum, with plant-growth-promoting and immune-stimulating properties towards Populus x canescens, our study revealed robustly accelerated growth towards phytopathogens, while showing a contrary response to ectomycorrhizal fungi. Transcriptomic analyses identified distinct genetic programs during interaction corresponding to the lifestyles, emphasizing the expression of mycoparasitism-related genes only in the presence of phytopathogens. Conclusion The findings reveal a critical mode of fungal community interactions belowground and suggest that Trichoderma spp. can distinguish between fungal partners of different lifestyles already at a distance. This sheds light on the entangled interactions of fungi in the rhizosphere and emphasizes the potential benefits of using Trichoderma spp. as a biocontrol agent and bio-fertilizer in tree plantations.

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]

The Canary Islands lead banana (Musa acuminata) production in the EU. Different fungal pathogens affect this crop in subtropical areas, with Fusarium oxysporum f. sp. cubense subtropical race 4 (Foc-STR4) being the most important in the Canary Islands. With the aim of developing environmentally sustainable techniques for disease control, this study presents the results of the evaluation of the antifungal capacity of a native Trichoderma collection (12 species, 109 isolates) obtained from banana soils. The results demonstrate the diversity of biocontrol genes and the in vitro antagonistic potential of different native Trichoderma species/isolates against two Foc-STR4 strains obtained from plants with Panama disease symptoms. Trichoderma virens (TF18), a dominant species in banana soils in the Canary Islands, showed a high capacity to inhibit the growth of Foc-STR4 in different in vitro assays. Trichoderma atrobrunneum (TF01) showed mycoparasitism capacity through the spiral coil around the hyphae of the pathogen. In addition, the genome analysis of T. atrobrunneum (TF03) showed 69 putative biosynthetic gene clusters, with the notable presence of the trichothecene tri5 gene. Finally, our work demonstrates that the soils of the Canary Islands banana crops are a potential source of environmentally adapted biological control agents to control or reduce the incidence of Foc-STR4. [ABSTRACT FROM AUTHOR]

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Pythium sensu stricto (s.s.) and Globisporangium species are important components of the soil microbial community and exhibit diverse lifestyles, including mycoparasitism. However, a comprehensive understanding of the species diversity of these mycoparasites in the West Azarbaijan province of Iran is lacking. In this study, a total of 114 mycoparasitic Pythium s.s. and Globisporangium isolates were obtained from agricultural soils collected from six regions in the province. Through DNA barcoding, all Globisporangium isolates were identified as G. nunn, while the barcode markers were insufficient to accurately resolve species boundaries in Pythium s.s. By combining morphological and multilocus sequence data, five species within the genus Pythium s.s. were identified: P. salmasense sp. nov., a potentially new species, and three known species, P. acanthicum, P. ornamentatum, and P. periplocum. Pythium ornamentatum was the most common species and found in all regions studied, followed by G. nunn and P. acanthicum, which were both isolated from four regions. While the isolates of G. nunn showed no mycoparasitic activity against Sclerotinia sclerotiorum, all Pythium s.s. species were capable of infecting the hyphae of this pathogen. The existence of mycoparasitic species is promising for biological control of soil‐borne fungal pathogens in the province. The widespread occurrence of P. ornamentatum, G. nunn, and P. acanthicum may suggest their adaptation to local soil and environmental conditions, indicating their potentially superior effectiveness in controlling plant diseases across different regions if used as biocontrol agents. [ABSTRACT FROM AUTHOR]

Trichoderma spp. are avirulent expedient plant symbionts. In addition, as plant-dependent organisms, Trichoderma species conjointly behave as a cost-effective and eco-friendly biocontrol agent. This review provides several mechanisms on how Trichoderma exerts useful effects on plants as a biocontrol agent. In addition to preventing disease, Trichoderma increases plant resistance and growth, improves nutrient uptake efficiency, and cleans the environment from pesticide contamination. Trichoderma spp. also functions as a safe, affordable, efficient, and environmentally beneficial biocontrol agent for various crops. The biological control mechanisms of Trichoderma, such as competition, antibiosis, antagonism, and mycoparasitism, as well as the mechanisms by which Trichoderma and plants promote plant growth and induce systemic resistance, have been discussed. In addition, the impact and applications of Trichoderma in treating different plant diseases are provided. Developing a diverse range of application technologies for Trichoderma is a crucial avenue for further research and development, given its potential to contribute to the sustainable growth of the agricultural sector. Nevertheless, due to its adaptable qualities, its entire potential cannot be fully realized. The knowledge of Trichoderma as a biocontrol agent, its secondary metabolites, and their use in plant disease management programs is explained, which suggests the potential use of Trichoderma as a safe, environmentally acceptable, and efficient biocontrol agent for several crop species. [ABSTRACT FROM AUTHOR]

Background: The coffee plant is a strategic crop in Saudi Arabia that makes a substantial contribution to the country’s economy. Therefore, it is crucial to continuously monitor and control the fungal phytopathogens that affect coffee plants in order to minimize crop losses and ensure sustainable cultivation. This is the first surveillance report of Fusarium phytopathogens associated with coffee plants in Saudi Arabia. Moreover, the antagonistic efficiency of the endophytic fungus, Diaporthe caatingaensis, was evaluated against the isolated Fusarium phytopathogens. Methods: The isolated strains were preliminary identified using cultural and microscopic methods and the identification was confirmed using internal transcribed spacer (ITS) sequencing technique. The detached leaf assay was conducted to assess the disease severity of Fusarium phytopathogens against detached coffee leaves. Moreover, dual culture assay was utilized to assess the antagonistic activity of D. caatingaensis. Results and conclusion: Fusarium oxysporum was found to be the most frequent isolated strain followed by F. solani, F. proliferatum and F. verticillioides strains. Fusarium proliferatum strain was found to be the most severe strain whereas F. solani strain showed the lowest disease severity. On the other hand, propiconazole fungicide was tested for its efficiency against Fusarium pathogens using food poisoning technique, showing that F. oxysporum strain 1 of accession number OP955665 was the most sensitive strain. However, F. proliferatum, F. oxysporum st. 2 (OP959851) and F. solani strains showed no significant response when the propiconazole concentration increase from 150 to 200 ppm. Scanning electron microscope proved the potent antagonistic activity of D. caatingaensis against F. proliferatum and F. oxysporum OP959874 strains through mycoparasitic modes of action as coiling, appressorium formation resulting in complete lysis of fungal mycelium. Accordingly, the current investigation provides the first surveillance data about Fusarium strains associated with coffee plants and also the utilization of D. caatingaensis as a potential biocontrol agent for effective management of Fusarium phytopathogens, avoiding the incidence of fungal resistance to fungicides and the harmful effects of commercial fungicides for sustainable cultivation of coffee in Saudi Arabia.

Trichoderma harzianum is a well-known biological control strain and a mycoparasite of Rhizoctonia solani. To explore the mechanisms of mycoparasitism, the genome and transcriptome of T. harzianum T4 were both assembled and analyzed in this study. The genome of T. harzianum T4 was assembled into 106 scaffolds, sized 41.25 Mb, and annotated with a total of 8118 predicted genes. We analyzed the transcriptome of T. harzianum T4 against R. solani in a dual culture in three culture periods: before contact (BC), during contact (C), and after contact (AC). Transcriptome sequencing identified 1092, 1222, and 2046 differentially expressed genes (DEGs), respectively. These DEGs, which are involved in pathogen recognition and signal transduction, hydrolase, transporters, antibiosis, and defense-related functional genes, are significantly upregulated in the mycoparasitism process. The results of genome and transcriptome analysis indicated that the mycoparasitism process of T. harzianum T4 was very complex. T. harzianum successfully recognizes and invades host cells and kills plant pathogens by regulating various DEGs at different culture periods. The relative expression levels of the 26 upregulated DEGs were confirmed by RT-qPCR to validate the reliability of the transcriptome data. The results provide insight into the molecular mechanisms underlying T. harzianum T4's mycoparasitic processes, and they provide a potential molecular target for the biological control mechanism of T. harzianum T4. [ABSTRACT FROM AUTHOR]

ABSTRACT Fungal cell walls are dynamic extracellular matrices that enable efficient adaptation to changing environments. While the cell wall compositions of yeasts, human, and plant pathogenic fungi have been studied to some extent, the cell walls of mycoparasites remain poorly characterized. Trichoderma species comprise a diverse group of soil fungi with different survival strategies and lifestyles. The comparative study of cell wall carbohydrate-active enzymes in 13 Trichoderma spp. revealed that the types of enzymes involved in chitin and chitosan metabolism are phylogenetically distant between mycoparasitic and saprotrophic species. Here, we compare the carbohydrate composition and function of the cell wall of a saprotrophic strain Trichoderma reesei with that of the mycoparasitic, biological control agent Trichoderma atroviride. Monosaccharide and glycosidic linkage analyses as well as dual in situ interaction assays showed that the cell wall polysaccharide composition is conserved between both species, except for the amounts of chitin detected. The results suggest that the observed accumulation of chitosan during mycoparasitism may prevent host recognition. Remarkably, Trichoderma atroviride undergoes dynamic cell wall adaptations during both vegetative development and mycoparasitism, which appears to be confirmed by an evolutionarily expanded group of specialized enzymes. Overall, our analyses support the notion that habitat specialization is reflected in cell wall architecture and that plastic chitin remodeling may confer an advantage to mycoparasites, ultimately enabling the successful invasion and parasitism of plant pathogens. This information may potentially be exploited for the control of crop diseases using biological agents.IMPORTANCETrichoderma species are emerging model fungi for the development of biocontrol agents and are used in industrial biotechnology as efficient enzyme producers. Fungal cell walls are complex structures that differ in carbohydrate, protein, and enzyme composition across taxa. Here, we present a chemical characterization of the cell walls of two Trichoderma spp., namely the predominantly saprotrophic Trichoderma reesei and the mycoparasite Trichoderma atroviride. Chemical profiling revealed that Trichoderma spp. remodel their cell wall to adapt to particular lifestyles, with dynamic changes during vegetative development. Importantly, we found that chitosan accumulation during mycoparasitism of a fungal host emerged as a sophisticated strategy underpinning an effective attack. These insights shed light on the molecular mechanisms that allow mycoparasites to overcome host defenses and can be exploited to improve the application of T. atroviride in biological pest control. Moreover, our results provide valuable information for targeting the fungal cell wall for therapeutic purposes.

The Pestalotiopsis sp. strain cr013 is a mycoparasite of Cronartium ribicola, a potential biocontrol fungus for Armand pine (Pinus armandii) blister rust. A previous study showed that the strain cr013 has great potential to produce new compounds. However, there has been no report of the whole-genome sequence of the mycoparasite Pestalotiopsis sp. In this study, the BGISEQ-500 and Oxford Nanopore GridION X5 sequencing platforms were used to sequence the strain cr013 isolates and assemble the reads to obtain the complete genome. We first report the whole-genome information of the mycoparasite Pestalotiopsis sp. strain cr013 (GenBank accession number: JACFXT010000000, BioProject ID: PRJNA647543, BioSample ID: SAMN15589943), and the genomic components and gene functions related to the mycoparasitism process were analyzed. This study provides a theoretical basis for understanding the lifestyle strategy of the mycoparasite Pestalotiopsis sp. and reveals the mechanisms underlying secondary metabolite diversity in the strain cr013.

The Canary Islands lead banana (Musa acuminata) production in the EU. Different fungal pathogens affect this crop in subtropical areas, with Fusarium oxysporum f. sp. cubense subtropical race 4 (Foc-STR4) being the most important in the Canary Islands. With the aim of developing environmentally sustainable techniques for disease control, this study presents the results of the evaluation of the antifungal capacity of a native Trichoderma collection (12 species, 109 isolates) obtained from banana soils. The results demonstrate the diversity of biocontrol genes and the in vitro antagonistic potential of different native Trichoderma species/isolates against two Foc-STR4 strains obtained from plants with Panama disease symptoms. Trichoderma virens (TF18), a dominant species in banana soils in the Canary Islands, showed a high capacity to inhibit the growth of Foc-STR4 in different in vitro assays. Trichoderma atrobrunneum (TF01) showed mycoparasitism capacity through the spiral coil around the hyphae of the pathogen. In addition, the genome analysis of T. atrobrunneum (TF03) showed 69 putative biosynthetic gene clusters, with the notable presence of the trichothecene tri5 gene. Finally, our work demonstrates that the soils of the Canary Islands banana crops are a potential source of environmentally adapted biological control agents to control or reduce the incidence of Foc-STR4.

Under natural conditions, crops typically suffer from severe challenges due to the increasing of abiotic and biotic stresses which severely affect plant growth and reduc crop yield. The present study investigated the single and combined impacts of Sclerotinia sclerotiorum and salinity stress on common bean (Phaseolus vulgaris L.) seedling which is scarcely studied. The study evaluated the in vitro and in vivo influence of two salinity tolerant Trichoderma isolates, T. koningii and T. harzianum against S. sclerotiorum under salinity stress. The results showed the ability of T. koningii and T. harzianum to grow and sporulate at high levels of salinity, 80 mM NaCl, without significantly impacting their ability to produce cell wall degrading enzymes, cellulase and chitinase. Amylase and proteinase (Prb1) genes were detected in T. harzianum. The in vitro assay revealed that both isolates could inhibit the growth of S. sclerotiorum under high salinity concentrations. In a greenhouse experiment, both Trichoderma isolates ameliorated the damaging impacts of S. sclerotiorum under salinity stress on common bean seedlings' germination and growth characteristics compared to their untreated control. Both bioagents significantly attenuated the damping-off and collar/stem rot percentages of infected common bean under salinity stress. Salinity stress intensified the effect of S. sclerotiorum on photosynthetic pigments, induced oxidative and nitrative stress, hampered ionic homeostasis, and deactivated antioxidants and defense-related molecules. On the other hand, Trichoderma isolates restrained the reduction of chlorophylls and carotenoids, ascorbate, reduced glutathione, flavonoids, phenolics, and various antioxidant enzymes, especially for single stresses and T. harzianum. All these upregulations reflected in keeping the cell membranes of common beans seedling more stable where the levels of lipid peroxidation and methylglyoxal due to the reduction of reactive oxygen species and upregulation of nitric oxide, which expressed better growth under pathogen attack or/and saline. The tested isolates, T. koningii and T. harzianum could be used as effective biological control against S. sclerotiorum on common beans in saline soils or areas irrigated with saline water.

ABSTRACTThe mycoparasitic fungus Trichoderma atroviride is applied in agriculture as a biostimulant and biologic control agent against fungal pathogens that infest crop plants. Secondary metabolites are among the main agents determining the strength and progress of the mycoparasitic attack. However, expression of most secondary metabolism-associated genes requires specific cues, as they are silent under routine laboratory conditions due to their maintenance in an inactive heterochromatin state. Therefore, histone modifications are crucial for the regulation of secondary metabolism. Here, we functionally investigated the role of the class II histone deacetylase encoding gene hda1 of T. atroviride by targeted gene deletion, phenotypic characterization, and multi-omics approaches. Deletion of hda1 did not result in obvious phenotypic alterations but led to an enhanced inhibitory activity of secreted metabolites and reduced mycoparasitic abilities of T. atroviride against the plant-pathogenic fungi Botrytis cinerea and Rhizoctonia solani. The ∆hda1 mutants emitted altered amounts of four volatile organic compounds along their development, produced different metabolite profiles upon growth in liquid culture, and showed a higher susceptibility to oxidative and osmotic stress. Moreover, hda1 deletion affected the expression of several notable gene categories such as polyketide synthases, transcription factors, and genes involved in the HOG MAPK pathway.IMPORTANCEHistone deacetylases play crucial roles in regulating chromatin structure and gene transcription. To date, classical—Zn2+ dependent—fungal histone deacetylases are divided into two classes, of which each comprises orthologues of the two sub-groups Rpd3 and Hos2 and Hda1 and Hos3 of yeast, respectively. However, the role of these chromatin remodelers in mycoparasitic fungi is poorly understood. In this study, we provide evidence that Hda1, the class II histone deacetylases of the mycoparasitic fungus Trichoderma atroviride, regulates its mycoparasitic activity, secondary metabolite biosynthesis, and osmotic and oxidative stress tolerance. The function of Hda1 in regulating bioactive metabolite production and mycoparasitism reveals the importance of chromatin-dependent regulation in the ability of T. atroviride to successfully control fungal plant pathogens.

The present study was aimed to test the antagonistic activity of endophytic fungi isolated from Aloe vera leaves against a number of plant-pathogenic fungi. The results showed that the highest antagonistic activity was exerted by Penicillium chermesinum against the pathogenic fungus Rhizoctonia solani, with an inhibition rate of 78.57%. Also, Talaromyces verruculosus and P. chermesinum showed their highest antagonistic activities against pathogenic fungus Fusarium 1 with inhibition rates of 61.79% and 61.53%, respectively, while T. verruculosus showed an inhibition rate of 62.91% against the pathogenic fungus Fusarium 2. Most of the endophytic fungi showed the least percentage of inhibition against Macrophomina phaseolina compared to the rest of the pathogenic fungi. The fungal filtrates showed a concentration-dependent inhibition rate against the plant pathogens, with a range of 0.00-48.13%. The highest inhibitory activity of Aspergillus niger was recorded at a concentration of 60% against Fusarium 2, R. solani, and Fusarium 1, with rates of 48.13, 37.39, and 36.66%, respectively. The results also showed the ineffectiveness of all fungal filtrates against the pathogenic fungus M. phaseolina. Chemical analysis of the filtrates of the endophytic fungi which showed antagonistic activity against pathogenic fungi was also conducted by using GC-MS. The results showed the presence of effective compounds with biological effects, such as Pentadecanoic acid, Oleic Acid, Limonene, cis-Vaccenic acid, Hexanedioic acid, bis(2-ethylhexyl) ester, Cycloheptasiloxane, and tetracamethyl-. [ABSTRACT FROM AUTHOR]

Trichoderma harzianum is a well-known biological control agent (BCA) that shows great potential in controlling many pathogenic fungi. To screen for genes associated with mycoparasitism, we sequenced and analyzed the transcriptome of T. harzianum T4 grown in dual culture with Colletotrichum musae. We analyzed differentially expressed genes (DEGs) of Trichoderma harzianum T4 in three different culture periods: before contact (BC), during contact (C) and after contact (AC). A total of 1453 genes were significantly differentially expressed compared to when T. harzianum T4 was cultured alone. During the three periods of double culture of T. harzianum T4 with C. musae, 74, 516, and 548 genes were up-regulated, respectively, and 11, 315, and 216 genes were down-regulated, respectively. The DEGs were screened using GO and KEGG enrichment analyses combined with differential expression multiples. Six gene categories related to mycoparasitism were screened: (a) pathogen recognition and signal transduction, (b) hydrolases, (c) ribosomal proteins and secreted proteins, (d) multidrug-resistant proteins and transporters, (e) heat shock proteins and detoxification, and (f) oxidative stress and antibiotics-related genes. The expression levels of 24 up-regulated genes during T. harzianum T4’s antagonistic interaction with C. musae were detected via real-time fluorescence quantitative PCR (RT-qPCR). This study provided information on the transcriptional expression of T. harzianum T4 against C. musae. These results may help us to further understand the mechanism of mycoparasitism, which can provide a potential molecular target for improving the biological control capacity of T. harzianum T4. [ABSTRACT FROM AUTHOR]

Four new species of the genus Niveomyces are described from Thailand. They were found as mycoparasites on: Ophiocordyceps infecting flies (Diptera) for Niveomyces albus; ants (Hymenoptera) for N. formicidarum; and leafhoppers (Hemiptera) for N. hirsutellae and N. multisynnematus. A new genus, Pseudoniveomyces with two species: Pseudoniveo. blattae (type species), parasitic on Ophiocordyceps infecting cockroaches, and Pseudoniveo. arachnovorum, found on a spider egg sac, are also described. These fungi share a common feature which is a sporothrix-like asexual morph. Based on our molecular data, Sporothrix insectorum is shown to be affiliated to the genus Niveomyces, and thus a new combination N. insectorum comb. nov. is proposed. Niveomyces coronatus, N. formicidarum and N. insectorum formed the N. coronatus species complex found on ant-pathogenic Ophiocordyceps from different continents. Pseudoniveomyces species are distinguished from Niveomyces spp. based on the presence of fusoid macroconidia in culture and a red pigment diffused in the medium, resembling to Gibellula and Hevansia. The molecular phylogenetic analyses also confirmed its generic status. The host/substrates associated with the genera within Cordycipitaceae were mapped onto the phylogeny to demonstrate that mycoparasitism also evolved independently multiple times in this family. [ABSTRACT FROM AUTHOR]

Claviceps purpurea causes ergot, a floral disease of major cereal crops, such as winter rye and winter triticale. The dark sclerotia formed by C. purpurea contain numerous toxic ergot alkaloids, which pose a major health risk for humans and animals when ingested. The countermeasures against ergot infection are currently limited to intensive tillage, seed cleaning, crop rotation or integration of optical sorting machines to remove ergot from cereals before processing. However, these practices confer only a minor protective effect and are highly energy-demanding. Thus, more effective and sustainable plant protection measures to combat ergot infections in cereals are needed. The application of Trichoderma spp. as biological control agent has already proven successful against a wide range of phytopathogenic fungi. However, there is only minor scientific evidence about its protective capacity against C. purpurea in cereal crops. In our study, we therefore investigated the antagonistic potential of several Trichoderma isolates against C. purpurea and their ability to confer protection against ergot infection. In initial in vitro tests and confrontation assays, we quantified and compared the production of iron-chelating compounds and hydrolytic enzymes as well as the antagonistic activity against C. purpurea sclerotia and mycelial cultures. Several strains showed high antifungal capacity and growth inhibitory effects towards C. purpurea. Selected Trichoderma strains were then tested for their potential to protect rye (Secale cereale) plants from ergot infection within a greenhouse trial. Rhizosphere-inoculated plants displayed a decreased disease severity compared to plants that had been sprayed with Trichoderma spores above-ground. Gene expression analysis by reverse transcription quantitative PCR also indicated the induction of systemic defense reactions. Overall, our data suggest that individual Trichoderma isolates possess a high antagonistic potential towards C. purpurea, which could be effective by direct mycoparasitism and by inducing systemic plant resistance, and therefore provide important guidance towards the development of Trichoderma treatments of cereals as biocontrol measure against ergot. [ABSTRACT FROM AUTHOR]

Abstract Sm1 and Chit42 of Trichoderma have been universally confirmed as crucial biocontrol factors against pathogen infection through induced resistance and mycoparasitism, respectively. However, not enough work has been conducted to understand the novel function of fused expression of these two proteins in Trichoderma. The results of this study demonstrated that Sm1-Chit42 protein (SCf) engineered T. afroharzianum strain OE:SCf exerted synergistic inhibition to Botrytis cinerea growth at multiple stages of mycoparasitic interaction of T. afroharzianum and B. cinerea including chemotropism sensing, hyphal coiling, hydrophobicity modulation, cell wall adhesion, virulence reduction and pathogen killing by ROS. These results highlight a novel mycoparasitic system in Trichoderma strains engineered with Sm1-Chit42 chimeric protein to combat B. cinerea growth and reproduction, which would lay a strong foundation for exploring a new engineered Trichoderma biofungicide created with chimeric proteins in the future.

There has been renewed interest in the application of endophytic fungi to control phytopathogenic fungi, which cause significant damage to crop health, ultimately leading to losses in agricultural productivity. Endophytic fungi inhibit pathogens via different modes of action—mycoparasitism, competition (for nutrients and ecological niches), antibiosis, and induction of plant defense—thus demonstrating the ability to control a wide range of phytopathogenic fungi in different growth phases and habitats. However, many studies have been conducted under laboratory conditions, and there is a huge lack of studies in which real field testing was performed. Aspergillus, Clonostachys, Coniothyrium, Trichoderma, and Verticillium have been proven to be the most effective fungal biocontrol agents. Trichoderma is regarded as the most promising group in commercial formulations. In this study, we attempted to emphasize the significance of fungal endophytes in controlling phytopathogenic fungi, while reporting recent advances in endophytic biology and application.

Trichoderma is a genus of filamentous fungi with several species widely studied and used as biological control agents (BCAs) in agriculture. With respect to fungal disease control, Trichoderma spp. can use different mechanisms of action, including mycoparasitism, antibiosis, competition and/or activation of plant defences. Armillaria spp. are the causal pathogens of Armillaria root rot (ARR) disease, which is widely distributed and of great economic and environmental importance in agriculture and forestry. ARR can be devastating to plant health, through colonisation and rotting of the root system and collar of the infected plant/tree, which can cause a reduction in yield and eventually death. Armillaria can live as a saprophyte that survives on dead plant material, making eradication of ARR extremely difficult. In an exhaustive analysis of work published to date on the control of Armillaria spp. by Trichoderma spp., using the WoS and Scopus databases, we identified 31 papers. We discuss these reports in light of their key findings, including number of publications per year, the countries where they were published, the journals and the citations obtained so far. Although the vast majority do not identify the mechanism of action involved in control, some describe mycoparasitism, antibiosis, competition or modification of the rhizospheric microbiota as being responsible for effective biological control of the pathogen. Further research into these mechanisms and additional in-depth analysis of less studied or currently unidentified mechanisms, such as the activation of plant defences, would be highly beneficial to our understanding of Trichoderma as a BCA of Armillaria.

ABSTRACT Serine proteases are a group of important hydrolytic enzymes that play vital roles in various cellular processes in fungi. In this study, the S8 serine protease-encoding gene CrKP43 was identified in the highly efficient Clonostachys chloroleuca 67–1 (formerly C. rosea 67–1) strain, which was markedly upregulated when parasitizing Sclerotinia sclerotiorum. The function of CrKP43 was investigated using gene deletion and complementation, and the results indicated that the lack of CrKP43 resulted in deformed fungal hyphae and cell morphology, inhibition of conidiation, and decreased antagonistic activity toward the pathogenic fungus Fusarium oxysporum f. sp. cucumerinum. Moreover, the mutants displayed much weaker mycoparasitic ability to S. sclerotiorum sclerotia and lower control efficiency against soybean Sclerotinia rot compared with the wild-type strain. All biological characteristics and biocontrol activities were recovered when the CrKP43 gene was reinserted into the fungus. Using qRT-PCR analysis and protein-protein interaction assays, it was further proved that the CrKP43 protein interacted with the mitogen-activated protein kinase (MAPK) Crmapk, suggesting serine proteases might be involved in the mycoparasitism of C. chloroleuca with the regulation of Crmapk. The findings improve our knowledge of serine proteases and their regulation in mycoparasites and help to illuminate the mechanisms underlying mycoparasitism of C. chloroleuca. IMPORTANCE Mycoparasites play important roles in the biocontrol of plant fungal diseases, during which they secret multiple hydrolases such as serine proteases to degrade their fungal hosts. In this study, we demonstrated that the serine protease CrKP43 was involved in C. chloroleuca development and mycoparasitism with the regulation of Crmapk. To the best of our knowledge, it is the first report on the functions and regulatory mechanisms of serine proteases in C. chloroleuca. Our findings will provide new insight into the regulatory mechanisms of serine proteases in mycoparasites and contribute to clarifying the mechanisms underlying mycoparasitism of C. chloroleuca, which will facilitate the development of highly efficient fungal biocontrol agents as well.

Verticillium dahliae is a soilborne fungal pathogen that causes vascular wilt diseases in a wide range of economically important crops, including eggplant. Trichoderma spp. are effective biological control agents that suppress a wide range of plant pathogens through a variety of mechanisms, including mycoparasitism. However, the molecular mechanisms of mycoparasitism of Trichoderma spp. in the degradation of microsclerotia of V. dahliae are not yet fully understood. In this study, the ability of 15 isolates of Trichoderma to degrade microsclerotia of V. dahliae was evaluated using a dual culture method. After 15 days, isolate HZA14 showed the greatest potential for microsclerotial degradation. The culture filtrate of isolate HZA14 also significantly inhibited the mycelial growth and conidia germination of V. dahliae at different dilutions. Moreover, this study showed that T. virens produced siderophores and indole-3-acetic acid (IAA). In disease control tests, T. virens HZA14 reduced disease severity in eggplant seedlings by up to 2.77%, resulting in a control efficacy of 96.59% at 30 days after inoculation. Additionally, inoculation with an HZA14 isolate increased stem and root length and fresh and dry weight, demonstrating plant growth promotion efficacy. To further investigate the mycoparasitism mechanism of T. virens HZA14, transcriptomics sequencing and real-time fluorescence quantitative PCR (RT-qPCR) were used to identify the differentially expressed genes (DEGs) of T. virens HZA14 at 3, 6, 9, 12, and 15 days of the interaction with microsclerotia of V. dahliae. In contrast to the control group, the mycoparasitic process of T. virens HZA14 exhibited differential gene expression, with 1197, 1758, 1936, and 1914 genes being up-regulated and 1191, 1963, 2050, and 2114 genes being down-regulated, respectively. Among these genes, enzymes associated with the degradation of microsclerotia, such as endochitinase A1, endochitinase 3, endo-1,3-beta-glucanase, alpha-N-acetylglucosaminidase, laccase-1, and peroxidase were predicted based on bioinformatics analysis. The RT-qPCR results confirmed the RNA-sequencing data, showing that the expression trend of the genes was consistent. These results provide important information for understanding molecular mechanisms of microsclerotial degradation and integrated management of Verticillium wilt in eggplant and other crops. [ABSTRACT FROM AUTHOR]

Trichoderma spp. has the ability to inhibit fungal plant pathogens through several mechanisms like the production of hydrolytic enzymes, mycoparasitism, coiling, and antibiosis and is therefore recommended as a potential and native biocontrol agent for effective control of soil-transmitted diseases. Various species of Trichoderma, like T. virens, T. asperellum, T. harzianum, etc., have been explored for their biocontrol activity against phytopathogens. There are different Trichoderma species and strains with respect to plant pathogens. Efforts have been made to develop effective and efficient methods, such as microencapsulation use of different polymers, adjuvants, or carriers, to increase the shelf-life and efficacy of Trichoderma formulations. The crucial aspects for the success of a biocontrol agent include developing and validating formulations, improvement in shelf-life, cost-effectiveness, easy accessibility, improved delivery systems, broad spectrum in action, robust performance (biocontrol), and integrative strategies for sustainable disease management. This review focuses on recent developments in the isolation, identification, preservation, substrates, consortium, quality control, mass production, delivery methods, field performance, registration, and commercialization of Trichoderma formulations for strategic development of next-generation multifunctional biological control formulations. [ABSTRACT FROM AUTHOR]

Trichoderma spp., is a highly efficient antagonist of root pathogens, such as Rhizoctonia solani, which causes loss in many crops. The aim of this research was to evaluate in vitro the antagonistic capacity of T. viride, T. koningii, T. harzianum and Trichoderma spp. isolates against R. solani from a potato crop. In confrontation tests, all Trichoderma isolates were classified as antagonists class 2 according to Bell scale, where T. harzianum and T. koningii showed more than 60% inhibition of the radial growth of R. solani at 120 h. In the interaction between T. harzianum and Trichoderma spp. with R. solani, as mycoparasitism strategy, vacuolization, lysis, coiling, and penetration were demonstrated, the last two were present in all Trichoderma isolates evaluated. [ABSTRACT FROM AUTHOR]

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

Sm1 and Chit42 of Trichoderma have been universally confirmed as crucial biocontrol factors against pathogen infection through induced resistance and mycoparasitism, respectively. However, not enough work has been conducted to understand the novel function of fused expression of these two proteins in Trichoderma. The results of this study demonstrated that Sm1-Chit42 protein (SCf) engineered T. afroharzianum strain OE:SCf exerted synergistic inhibition to Botrytis cinerea growth at multiple stages of mycoparasitic interaction of T. afroharzianum and B. cinerea including chemotropism sensing, hyphal coiling, hydrophobicity modulation, cell wall adhesion, virulence reduction and pathogen killing by ROS. These results highlight a novel mycoparasitic system in Trichoderma strains engineered with Sm1-Chit42 chimeric protein to combat B. cinerea growth and reproduction, which would lay a strong foundation for exploring a new engineered Trichoderma biofungicide created with chimeric proteins in the future. [ABSTRACT FROM AUTHOR]

Clonostachys rosea is an important mycoparasitism biocontrol agent that exhibits excellent control efficacy against numerous fungal plant pathogens. Transcriptomic sequencing may be used to preliminarily screen mycoparasitism-related genes of C. rosea against fungal pathogens. The present study sequenced and analyzed the transcriptome of C. rosea mycoparasitizing a Basidiomycota (phylum) fungal pathogen, Rhizoctonia solani, under three touch stages: the pre-touch stage, touch stage and after-touch stage. The results showed that a number of genes were differentially expressed during C. rosea mycoparasitization of R. solani. At the pre-touch stage, 154 and 315 genes were up- and down-regulated, respectively. At the touch stage, the numbers of up- and down-regulated differentially expressed genes (DEGs) were 163 and 188, respectively. The after-touch stage obtained the highest number of DEGs, with 412 and 326 DEGs being up- and down-regulated, respectively. Among these DEGs, ABC transporter-, glucanase- and chitinase-encoding genes were selected as potential mycoparasitic genes according to a phylogenetic analysis. A comparative transcriptomic analysis between C. rosea mycoparasitizing R. solani and Sclerotinia sclerotiorum showed that several DEGs, including the tartrate transporter, SDR family oxidoreductase, metallophosphoesterase, gluconate 5-dehydrogenase and pyruvate carboxylase, were uniquely expressed in C. rosea mycoparasitizing R. solani. These results significantly expand our knowledge of mycoparasitism-related genes in C. rosea and elucidate the mycoparasitism mechanism of C. rosea. [ABSTRACT FROM AUTHOR]

Clonostachys chloroleuca (formerly classified as C. rosea) is an important mycoparasite active against various plant fungal pathogens. Mitogen-activated protein kinase (MAPK) signaling pathways are vital in mycoparasitic interactions; they participate in responses to diverse stresses and mediate fungal development. In previous studies, the MAPK-encoding gene Crmapk has been proven to be involved in mycoparasitism and the biocontrol processes of C. chloroleuca, but its regulatory mechanisms remain unclear. Aldose 1-epimerases are key enzymes in filamentous fungi that generate energy for fungal growth and development. By protein–protein interaction assays, the glucose-6-phosphate 1-epimerase CrGlu6 was found to interact with Crmapk, and expression of the CrGlu6 gene was significantly upregulated when C. chloroleuca colonized Sclerotinia sclerotiorum sclerotia. Gene deletion and complementation analyses showed that CrGlu6 deficiency caused abnormal morphology of hyphae and cells, and greatly reduced conidiation. Moreover, deletion mutants presented much lower antifungal activities and mycoparasitic ability, and control efficiency against sclerotinia stem rot was markedly decreased. When the CrGlu6 gene was reinserted, all biological characteristics and biocontrol activities were recovered. These findings provide new insight into the mechanisms of glucose-6-phosphate 1-epimerase in mycoparasitism and help to further reveal the regulation of MAPK and its interacting proteins in the biocontrol of C. chloroleuca. [ABSTRACT FROM AUTHOR]

Sclerotinia sclerotiorum is an important plant pathogenic fungus of many crops. Our previous study identified the S. sclerotiorum agglutinin (SSA) that can be partially degraded by the serine protease CmSp1 from the mycoparasite Coniothyrium minitans. However, the biological functions of SSA in the pathogenicity of S. sclerotiorum and in its response to infection by C. minitans, as well as to environmental stresses, remain unknown. In this study, SSA disruption and complementary mutants were generated for characterization of its biological functions. Both the wild-type (WT) of S. sclerotiorum and the mutants were compared for growth and sclerotial formation on potato dextrose agar (PDA) and autoclaved carrot slices (ACS), for pathogenicity on oilseed rape, as well as for susceptibility to chemical stresses (NaCl, KCl, CaCl2, sorbitol, mannitol, sucrose, sodium dodecyl sulfate, H2O2) and to the mycoparasitism of C. minitans. The disruption mutants (ΔSSA-175, ΔSSA-178, ΔSSA-225) did not differ from the WT and the complementary mutant ΔSSA-178C in mycelial growth. However, compared to the WT and ΔSSA-178C, the disruption mutants formed immature sclerotia on PDA, and produced less but larger sclerotia on ACS; they became less sensitive to the eight investigated chemical stresses, but more aggressive in infecting leaves of oilseed rape, and more susceptible to mycoparasitism by C. minitans. These results suggest that SSA positively regulates sclerotial development and resistance to C. minitans mycoparasitism, but negatively regulates pathogenicity and resistance to chemical stresses. [ABSTRACT FROM AUTHOR]

Fungi of the Trichoderma genus have been widely studied for their mycoparasitic characteristics that allow the generation of biological control products. In addition, some of them promote plant growth. Currently, there are over 300 species divided into 8 clades or lineages. Despite such diversity, few species have been reported in Mexico, and there is not representativeness of every clade. Here, we present the identification of the 2IG2102 strain of T. brevicompactum based on analysis of 3 phylogenetic markers, as well as its characterization as antagonist towards 2 Fusarium fungi, and it also boosts tomato and cucumber seed germination. Trichothecene biosynthesis might be a mechanism required to inhibit phytopathogens growth and in parallel affect the post-germinative growth of tomato. This is the first report of a Trichoderma belonging to the clade 6 isolated in Mexico, that displays capacity to antagonize fungi and it might produce trichothecenes for biological control. [ABSTRACT FROM AUTHOR]

Pathogenic root/wood rot fungal species infect multiple urban tree species in Singapore. There is a need for sustainable and environmentally friendly mitigation. We report the local Trichoderma strains as potential biocontrol agents (BCAs) for pathogenic wood rot fungal species such as Phellinus noxius, Rigidoporus microporus, and Fulvifomes siamensis. Isolated Trichoderma strains were DNA-barcoded for their molecular identities and assessed for their potential as a BCA by their rate of growth in culture and effectiveness in inhibiting the pathogenic fungi in in vitro dual culture assays. Trichoderma harzianum strain CE92 was the most effective in inhibiting the growth of the pathogenic fungi tested. Preliminary results suggested both volatile organic compound (VOC) production and direct hyphal contact contributed to inhibition. SPME GC-MS identified known fungal inhibitory volatiles. Trichoderma harzianum strain CE92 hyphae were found to coil around Phellinus noxius and Lasiodiplodia theobromae upon contact in vitro and were possibly a part of the mycoparasitism. In summary, the work provides insight into Trichoderma inhibition of pathogenic fungi and identifies local strains with good potential for broad-spectrum BCAs against root/wood rot fungi in Singapore. [ABSTRACT FROM AUTHOR]

Trichoderma isolates were inhibited variably in-vitro growth of soil-borne phytopathogen Macrophomina phaseolina (Maubl.) Ashby causes root rot in cotton. The growth inhibition of test-pathogen was found to be higher (90.36%) in T. viride NBAIITv23 followed by T. koningii MTCC796 (85.77%) under dual culture antagonism. The microscopic examination suggested that the antagonists Tv23 and MTCC796 adopted mycoparasitism as a strong mode of action to restrain pathogen growth. However, antagonists T. harzianum NBAIITh1 (77.89%) and T. virens NBAIITvs12 (61.74%) demonstrated strong antibiosis action for growth inhibition of the test pathogen. A significant positive correlation was established between the growth inhibition of M. phaseolina and the release of cell wall degrading enzymes- chitinase (p = 0.001), β-1,3, glucanase (p = 0.01), and protease (p = 0.05) under the influence of pathogen cell wall. The chitinase and β-1,3, glucanase activities were elevated 2.09 and 1.75 folds, respectively, in potent mycoparasitic Tv23 strain influenced by a pathogen cell wall compared to glucose as a carbon source. The three unique DNA-RAPD fragments OPA-07(1033), OPA-16(983), and OPO-15(239), amplified by potent mycoparasitic Tv23 strain, were subjected to DNA sequencing and derived functional 864 bp from OPA-16(983) and have sequence homology to ech42 gene with partial CDs of 262 amino acids (nucleotide accession No. KF723016.1 and protein accession No.AHF57046.1). Novel SCAR markers were developed from a functional sequence of OPA-16 fragments and validated across the genomic DNA of eleven Trichoderma antagonists. The novel SCAR markers evolved from the RAPD-SCAR interface to authenticate chitinolytic Trichoderma associated with mycoparasitic action for eco-friendly biocontrol activity. [ABSTRACT FROM AUTHOR]

Clonostachys (Bionectriaceae, Hypocreales) species are common soil-borne fungi, endophytes, epiphytes, and saprotrophs. Sexual morphs of Clonostachys spp. were placed in the genus Bionectria, which was further segregated into the six subgenera Astromata, Bionectria, Epiphloea, Myronectria, Uniparietina, and Zebrinella. However, with the end of dual nomenclature, Clonostachys became the single depository for sexual and asexual morph- typified species. Species of Clonostachys are typically characterised by penicillate, sporodochial, and, in many cases, dimorphic conidiophores (primary and secondary conidiophores). Primary conidiophores are mononematous, either verticillium-like or narrowly penicillate. The secondary conidiophores generally form imbricate conidial chains that can collapse to slimy masses, particularly on sporodochia. In the present study, we investigated the species diversity within a collection of 420 strains of Clonostachys from the culture collection of, and personal collections at, the Westerdijk Fungal Biodiversity Institute in Utrecht, the Netherlands. Strains were analysed based on their morphological characters and molecular phylogeny. The latter used DNA sequence data of the nuclear ribosomal internal transcribed spacer regions and intervening 5.8S nrDNA (ITS) and partial 28S large subunit (LSU) nrDNA and partial protein encoding genes including the RNA polymerase II second largest subunit (RPB2), translation elongation factor 1-alpha (TEF1) and β-tubulin (TUB2). Based on these results, the subgenera Astromata, Bionectria, Myronectria and Zebrinella are supported within Clonostachys. Furthermore, the genus Sesquicillium is resurrected to accommodate the former subgenera Epiphloea and Uniparietina. The close relationship of Clonostachys and Sesquicillium is strongly supported as both are inferred phylogenetically as sister-genera. New taxa include 24 new species and 10 new combinations. Recognition of Sesquicillium distinguishes species typically forming a reduced perithecial stroma superficially on plant tissue from species in Clonostachys often forming well-developed, through bark erumpent stromata. The patterns of observed perithecial wall anatomies, perithecial wall and stroma interfaces, and asexual morph diversifications described in a previously compiled monograph are used for interpreting ancestral state reconstructions. It is inferred that the common ancestor of Clonostachys and Sesquicillium may have formed perithecia superficially on leaves, possessed a perithecial wall consisting of a single region, and formed intercalary phialides in penicilli of conidiophores. Character interpretation may also allow hypothesising that diversification of morphs occurred then in the two genera independently and that the frequently stroma-linked Clonostachys morphs evolved together with the occupation of woody host niches and mycoparasitism. [ABSTRACT FROM AUTHOR]

There has been renewed interest in the application of endophytic fungi to control phytopathogenic fungi, which cause significant damage to crop health, ultimately leading to losses in agricultural productivity. Endophytic fungi inhibit pathogens via different modes of action—mycoparasitism, competition (for nutrients and ecological niches), antibiosis, and induction of plant defense—thus demonstrating the ability to control a wide range of phytopathogenic fungi in different growth phases and habitats. However, many studies have been conducted under laboratory conditions, and there is a huge lack of studies in which real field testing was performed. Aspergillus, Clonostachys, Coniothyrium, Trichoderma, and Verticillium have been proven to be the most effective fungal biocontrol agents. Trichoderma is regarded as the most promising group in commercial formulations. In this study, we attempted to emphasize the significance of fungal endophytes in controlling phytopathogenic fungi, while reporting recent advances in endophytic biology and application. [ABSTRACT FROM AUTHOR]

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

Phallus rubrovolvatus is a unique mushroom used for medicinal and dietary purposes in China. In recent years, however, the rot disease of P. rubrovolvatus has seriously affected its yield and quality, becoming an economically important threat. In this study, samples of symptomatic tissues were collected, isolated, and identified from five major P. rubrovolvatus production regions in Guizhou Province, China. Based on combined analyses of phylogenies (ITS and EF1-α), morphological characteristics and Koch's postulates, Trichoderma koningiopsis and Trichoderma koningii were identified as the pathogenic fungal species. Among these, T. koningii exhibited stronger pathogenicity than the other strains; thus, T. koningii was used as the test strain in the follow-up experiments. Upon co-culturing T. koningii with P. rubrovolvatus, the hyphae of the two species were intertwined, and the color of the P. rubrovolvatus hyphae changed from white to red. Moreover, T. koningii hyphae were wrapped around P. rubrovolvatus hyphae, leading to their shortening and convolution and ultimately inhibiting their growth due to wrinkling; T. koningii penetrated the entire basidiocarp tissue of P. rubrovolvatus, causing serious damage to the host basidiocarp cells. Further analyses revealed that T. koningii infection resulted in the swelling of basidiocarps and significantly enhanced the activity of defense-related enzymes, such as malondialdehyde, manganese peroxidase, and polyphenol oxidase. These findings offer theoretical support for further research on the infection mechanisms of pathogenic fungi and the prevention of diseases caused by them. [ABSTRACT FROM AUTHOR]