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Green mould contamination causes a significant challenge to mushroom growers in Malaysia leading to reduced yields and economic losses in the widely cultivated and marketed edible grey oyster mushroom, Pleurotus pulmanorius. This study aimed to identify the causal agents of green mould contaminants and determine the critical points in the cultivation process in the farm that contribute to green mould contamination. Samples of mushroom substrate (sawdust), spawn substrate (corn), environmental sources and tools were collected at different stages of mushroom cultivation. As results, the causal agents of green mould contamination were identified as Trichoderma pleuroti , T. harzianum and T. ghanese. Prior to steam pasteurisation and after steam pasteurisation, the spawn substrate and mushroom substrate were found to be free of Trichoderma. However, Trichoderma was detected in water, air within the production house and on cleaning tools. This findings suggests that water could serve as the source of green mould introduction in mushroom farms, while cultivation practices such as watering and scratching during the harvesting cycle may contribute to adverse green mould. Understanding these critical points and causal agents provides information to mitigate the green mould contamination throughout the grey oyster mushroom cultivation process. [Display omitted] • Pleurotus mushroom cultivation in Malaysia faces contamination by green mould. • T. pleuroti is the main causal agent followed by T. harzianum and T. ghanense. • Water is the primary source followed by air inside the production house. • Watering is one of critical points in mushroom cultivation. [ABSTRACT FROM AUTHOR]

• Transgenerational immune priming in wheat facilitated by Trichoderma demonstrates long-lasting, broad-spectrum protection against biotic stresses. • Inheritance of priming improves disease resistance, biochemical responses, and yield-related traits. • Primed wheat shows clear superiority over non-primed counterparts. • Ability to mount targeted defense response conserves resources and enhances yield under biotic stress. • Inheritance of Trichoderma -mediated priming offers significant breakthrough for sustainable agriculture as non-chemical crop protection alternative. Plants encounter various biotic stresses in their natural habitat, prompting the exploration of innovative protective measures to bolster their immune defences against pathogens. An effective strategy involves priming plant immunity, conferring enduring and comprehensive protection. Defence priming, characterized by heightened responsiveness to attackers following prior stress exposure, intriguingly manifests transgenerationally, transmitting the memory of priming to subsequent plant generations. In our investigation, we delved into the phenomenon of transgenerational immune priming (TGIP) in wheat facilitated by Trichoderma , spanning from the parental generation (G0) to the grand progeny (G2) generation. Successful establishment of transgenerational priming in the G2 generation was achieved by initially priming the G0 generation of wheat with Trichoderma. To assess priming inheritance, seeds from the G1 generation were collected, and both primed and non-primed wheat plants' grand progeny (G2 generation) were cultivated in soil. Subsequent evaluation encompassed the disease phenotype, biochemical parameters, and yield-related traits of the G2 generation wheat when confronted with Bipolaris sorokiniana , the causal agent of spot blotch disease. Our analysis revealed that the G2 generation of primed wheat displayed superior protection against spot blotch compared to non-primed wheat. Biochemical studies indicated no activation of defensive responses in the absence of disease pressure in G2 wheat plants. However, upon pathogen challenge, the grand progeny of primed wheat exhibited a robust defence response, surpassing that of non-primed wheat. This targeted defence response enabled efficient resource utilization and mitigated yield penalties. Furthermore, under biotic stress conditions, the grand progeny of primed wheat exhibited enhanced yield parameters compared to non-primed wheat. Our findings provide conclusive evidence that Trichoderma -mediated priming against B. sorokiniana can be inherited successfully from the parental (G0) generation to the grand progeny (G2) generation of wheat. This study represents the initial documentation of Trichoderma -mediated TGIP in wheat against B. sorokiniana , holding significant promise for sustainable agriculture by offering an environmentally friendly alternative to chemical pesticides. [ABSTRACT FROM AUTHOR]

Volatile organic compounds (VOCs) produced by yeasts can positively affect crops, acting as antifungals or biostimulants. In this study, Aureobasidium pullulans and Metschnikowia pulcherrima were evaluated as potential antagonists of Trichoderma spp., common fungal pathogen in mushroom cultivation. To assess the biocontrol ability and biostimulant properties of the selected yeast species, in vitro co-culture and VOCs exposure assays were conducted. In both assays, VOCs produced by Aureobasidium spp. showed the stronger antifungal activity with a growth inhibition up to 30 %. This result was further confirmed by the higher volatilome alcohol content revealed by solid phase microextraction-gas chromatography mass spectrometry (SPME/GC–MS). Overall, Aureobasidium strains can be potentially used as biocontrol agent in Pleorotus ostreatus and Cyclocybe cylindracea mycelial growth, without affecting their development as demonstrated by VOCs exposure assay and Fourier-transform infrared spectroscopy (FT-IR). Conversely, M. pulcherrima was characterized by a lower or absent antifungal properties and by a volatilome composition rich in isobutyl acetate, an ester often recognized as plant growth promoter. As confirmed by FT-IR, Lentinula mycelia exposed to M. pulcherrima VOCs showed a higher content of proteins and lipids, suggesting an improvement of some biochemical properties. Our study emphasizes that VOCs produced by specific yeast strains are potentially powerful alternative to synthetic fungicide in the vegetative growth of mushroom-forming fungi and also able to modify their biochemical composition. • Yeasts VOCs are characterized by different antifungal and biostimulant properties. • Aureobasidium spp. VOC have the stronger antifungal activity against Trichoderma spp. • M. pulcherrima VOCs improve mushrooms growth and nutritional characteristics. • FT-IR spectra suggested M. pulcherrima VOCs as biostimulant of Lentinula edodes. [ABSTRACT FROM AUTHOR]

The use of beneficial organisms for the biocontrol of soil-borne pathogens in forestry is still poor explored. In this work, the nutritional demands of 10 previously selected isolates of Trichoderma for the biocontrol of forest soil-borne pathogens have been tested by Phenotype Microarray technology, to investigate about their C-source utilization and exploring the possibility to obtain a microbial consortia (SynCom), an innovative strategy for the biocontrol of plant disease. All Trichoderma isolates tested in this study showed a high spore germination percentage within 3 d and evidenced nutritional preference regardless of the species they belong to, and unrelated to their soil of origin. Results of growth curve analysis and MANOVA test revealed that all isolates assimilate a broad range of substrates, generally preferring complex compounds such as monosaccharides related compounds, nitrogen compounds, carboxylic acids and esters. No evidence of competition for nutritional resources have been observed among isolates of this study. As a result, a combination of different isolates could be proposed to obtain a SynCom useful for the practice of phytopathogen biocontrol in forestry. The addition of i-erythritol, adenosine and turanose to a growth substrate could be suggested as stimulating compounds for the growth of the selected Trichoderma isolates. • Nutrient assimilation of microbial strains can be examined by phenotype microarray. • The tested Trichoderma isolates don't compete for any of the tested C-sources. • Turanose, i-erythritol and adenosine stimulate the growth of Trichoderma hamatum and Trichoderma koningiopsis tested isolates. [ABSTRACT FROM AUTHOR]

• This is the first study that successfully expressed genes encoding chitinase 42 kDa from Trichoderma asperellum SH16 in peanuts (Arachis hypogaea) cultivar L14 through Agrobacterium tumefaciens -mediated transformation. • Two synthetic genes (syncodChi42–1 and syncodChi42–2) optimized for codon usage for plant expression proved to be more efficient than their original gene (Chi42) from T. asperellum SH16 in transgenic peanuts. • The high expression level of two synthetic genes in peanut roots as well as their strong resistance to Sclerotium rolfsii promise potential applications in the field of transgenic crops against phytopathogenic fungi. This study aims to extracellularly express the 42 kDa chitinase from Trichoderma asperellum SH16 in peanut (Arachis hypogaea) roots under the regulation of a tissue-specific Asy promoter and the driving of the signal peptide of the rice amylase 3D gene. The 42 kDa chitinase genes were employed in this study, which included one wild-type sequence (Chi42) and two synthetic sequences (syncodChi42–1 and syncodChi42–2) optimized for codon usage for plant expression. From 102 42 kDa chitinase transgenic peanut lines, seven elite lines were discovered, among them two for syncodChi42–1, three for syncodChi42–2, and two for Chi42. The study also revealed chitinase from two synthetic genes had higher expression levels than chitinase from the wild-type gene, with the highest specific activities of approximately 901 U/mg and 1124 U/mg, respectively, compared to 739 U/mg. All seven elite transgenic peanut lines exhibited strong extracellular chitinase activity, and some of them were able to fight the soil-borne pathogenic fungus Sclerotium rolfsii in both in vitro and in vivo assays. The significant chitinase activity of seven elite transgenic peanut lines, particularly five lines carrying one of two synthetic genes, suggests that transgenic crops might be used to prevent phytopathogenic fungi. [ABSTRACT FROM AUTHOR]

[Display omitted] • Trichoderma species from soil and plants are rich source of bioactive secondary metabolites. • A total of 253 terpenoids with diverse skeletons and bioactivities were isolated from Trichoderma species. • The promising biosynthetic potential of Trichoderma species for terpenoid diversity were generalized and discussed. • Trichoderma species are of great significance for exploring drug lead compounds and potential application in agriculture. Trichoderma species are rich source of bioactive secondary metabolites. In the past decades, a series of secondary metabolites were reported from different Trichoderma fungi, among which terpenoids possessing versatile structural diversities and extensive pharmacological activities are one of the particularly important categories. The review aims to summarize the terpenoids isolated from Trichoderma species regarding their structural diversities, biological activities, and promising biosynthetic potentials. So far, a total of 253 terpenoids, including 202 sesquiterpenes, 48 diterpenes, 2 monoterpenes and 1 meroterpenoid, were isolated and identified from Trichoderma species between 1948 and 2022. Pharmacological investigations of Trichoderma terpenoids mainly focused on their antibacterial activities, antifungal activities, inhibitory activities on marine plankton species and cytotoxic activities, indicating that Trichoderma species are important microbial agents for drug discovery and environmentally friendly agrochemicals development. Intriguing chemistry and enzymology involved in the biosynthesis of Trichoderma terpenoids were also presented to facilitate further precise genome mining-guided novel structure discovery. Taken together, the abundance of novel skeletons, bioactivities and biosynthetic potentials presents new opportunities for drug and agrochemicals discovery, genome mining and enzymology exploration from Trichoderma species. The work will provide references for the profound study of terpenoids derived from Trichoderma , and facilitate further studies on Trichoderma species in the areas of chemistry, medicine, agriculture and microbiology. [ABSTRACT FROM AUTHOR]

All fungi emit mixtures of volatile organic compounds (VOCs) during growth. The qualitative and quantitative composition of these volatile mixtures vary with the species of fungus, the age of the fungus, and the environmental parameters attending growth. In nature, fungal VOCs are found as combinations of alcohols, aldehydes, acids, ethers, esters, ketones, terpenes, thiols and their derivatives, and are responsible for the characteristic odors associated with molds, mushrooms and yeasts. One of the single most common fungal volatiles is 1-octen-3-ol also known as "mushroom alcohol" or "matsutake alcohol." Many volatiles, including 1-octen-3-ol, serve as communication agents and display biological activity as germination inhibitors, plant growth retardants or promoters, and as semiochemicals ("infochemicals") in interactions with arthropods. Volatiles are understudied and underappreciated elements of the chemical lives of fungi. This review gives a brief introduction to fungal volatiles in hopes of raising awareness of the physiological importance of these gas phase fungal metabolites to encourage mycologists and other biologists to stop "throwing away the head space." • Filamentous fungi and yeasts are prolific producers of volatile organic compounds (VOCs). • Fungal VOCs are responsible for the characteristic odors associated with molds, mushrooms, and yeasts. • Molds growing in water damaged buildings release VOCs that may cause risks to human health. • Fungal VOCs can affect plant growth in positive and negative ways. • Trichoderma VOCs inhibit the growth of certain plant-pathogenic fungi and oomycetes. [ABSTRACT FROM AUTHOR]

Neofusicoccum parvum , is a fungal pathogen and one of the etiological agents of dieback disease in grapevines. The fungus causes deterioration of vines due to vascular colonization and/or production of toxins. We report herein the inhibitory effects of Trichoderma spp. isolates and the antifungal effects of cell-free supernatants (CFS) from Xenorhabdus and Photorhabdus bacteria against N. parvum in agar plates. We also evaluated the effects of the most effective fungi and bacteria against the pathogen in pruning wounds of vine shoots. All isolates of Trichoderma exhibited antifungal activity ranging between 82 and 97.5% at 14 days of post-treatment. All Xenorhabdus and Photorhabdus CFS at 10 and 33% concentrations inhibited mycelial growth with X. szentirmaii PAM 11 and PAM 25 causing the highest inhibition (>74%). In the shoot experiments, T. asperellum IB 01/13 and T. asperellum Quality®, X. szentirmaii PAM 11 (undiluted growth culture and CFS) suppressed the fungus by ≥ 93%. Our study highlights the potential of Trichoderma and X. szentirmaii PAM 11 for use as biofungicides in the management of N. parvum in grapevines. Further studies should be conducted to develop formulations of Trichoderma and Xenorhabdus that enhance stability in shelf-life and increase the efficacy of N. parvum control in grapevines under field conditions. [Display omitted] • Fungal diseases cause serious problems in grape production. • Neofusicoccum parvum is a fungal agent of dieback disease in grapevines. • Trichoderma is antagonistic against N. parvum in vines. • Xenorhabdus spp. produce putative biofungicide/s against dieback in vines. [ABSTRACT FROM AUTHOR]

Endochitinase is a natural extracellular protein in Trichoderma longibrachiatum T6, which can degrade the eggshell of Heterodera avenae significantly, however the related genes that coding this protein was rarely characterized. In the present study, the endochitinase 18-5 gene (T6-Echi18-5) of T. longibrachiatum T6 was cloned and sequenced. The expression level of T6-Echi18-5 gene in T. longibrachiatum T6 was induced and increased after the H. avenae cysts inoculation. The full-length cDNA sequence of T6-Echi18-5 was 1671 bp that contained an ORF of 1275 bp, corresponding to 424 amino acids with a 45.9 kDa molecular weight. A single band of 60.04 kDa was detected and identified using SDS-PAGE and Western blot analysis after transferring the T6-Echi18-5 gene to Escherichia coli BL21 Rosetta (DE3). The concentration of purified recombinant T6-Echi18-5 protein was 1.53 mg·ml−1, and the optimal temperature and pH were 50 °C and 5.0, respectively. The eggshell and content were dissolved and exuded from 4 to10 days after treatment with the purified recombinant T6-Echi18-5 protein. The relative inhibition rate of eggs hatching was 86.79 % at 12 days after treatment. Our study demonstrated the key role of T6-Echi18-5 gene in degrading the H. avenae eggshell and inhibiting the eggs hatching. • T6-Echi18-5 protein is a promising biological macromolecule for control H. avenae. • A nematocidal activity of endochitinase 18-5 gene was explored and sequenced. • The expression level of T6-Echi18-5 gene increased after induction with cysts. • T6-Echi18-5 protein exhibited high nematicidal activity and eggshell degradation. [ABSTRACT FROM AUTHOR]

The fungal disease of Cordyceps militaris has become the primary disease during the cultivation of C. militaris. However, some of the causal organism remains unclear. During our investigation in 2020–2022, three fungal strains were isolated from both the mycelium and fruiting bodies of C. militaris in mushroom farms located in Changchun. The observed disease incidence was approximately 20%. White villous mycelium on the surface of hyphae or fruiting bodies of C. militaris , is one of the main features of the disease, which can cause slow growth of mycelium, delayed primordia differentiation, and growth failure. Finally, the white cotton flocculent mycelium covered the surface of the medium, and the C. militaris stopped differentiating and failed to produce the primordium, ultimately leading to the stunted growth and lodging of the fruiting body. The objective of this study was to identify the pathogen based on Koch's Postulates, thereby confirming its pathogenicity and characterizing it through morphological and molecular features. The phylogenetic analyses showed that they belong to the Trichoderma gamsii. This study presented the first evidence of T. gamsii caused fungal disease in C. militaris , providing a theoretical basis for the prevention and control measures. • The outbreak of fungal diseases in Cordyceps militaris cultivation poses a significant threat to production and quality. • The disease incidence was approximately 20%, which led to significant yield losses. • The pathogen responsible for white mildew disease was identified as Trichoderma gamsii. • This is the first report of Trichoderma gamsii causing white mildew disease in Cordyceps militaris in China. • The results facilitate the development of strategies for white mildew disease of Cordyceps militaris. [ABSTRACT FROM AUTHOR]

The continuous monocropping obstacle, resulting from imbalance of nutrients and soil microorganisms, is the main factor limiting the productivity and quality of Pseudostellaria heterophylla (Miq.) Pax ex Pax et Hoffm. However, few agricultural methods and products are available to increase the productivity and quality of P. heterophylla by regulation the soil nutrients and microorganisms in continuous monocropping fields. In this study, Trichoderma consortium compost corncob (TCCC) was applied to P. heterophylla under continuous monocropping conditions to investigate its effects on growth, productivity, quality, disease incidence and to explore its impact on the soil nutrients and microbial communities. TCCC significantly enhanced the number of fibrous roots, the number of tuberous roots, the weight of 50 tuberous roots, the yield, and the drying rate of tuberous roots by 93.78–186.68%, 48.48–51.52%, 22.05–39.69%, 40.26–117.41%, and 4.23–6.67%, respectively. In addition, it significantly enhanced the crude polysaccharides content by 22.08–30.17% in P. heterophylla , whereas it significantly decreased the total saponin content by 20.00–22.50%. TCCC significantly lowered the incidence of leaf spot and root rot diseases by 69.71–73.86% and 18.64–53.25%, respectively. Moreover, applying TCCC significantly enhanced organic matter (OM), total nitrogen (TN), available nitrogen (AN), and total phosphorus (TP). Finally, the microbial analysis showed that TCCC significantly increased the relative abundance of beneficial microbiota (i.e., Trichoderma , Gaiella , and Chaetomium), while the relative abundances of pathogens such as Fusarium , Gibberella , Clonostachys , and Neocosmospora significantly decreased. Correlation analysis and structural equation model demonstrated that TCCC may promote the growth and productivity of P. heterophylla and alleviate its disease incidence under continuous monocropping conditions via reshaping the soil microbial community and enhancing soil nutrients. The present findings suggest that applying TCCC represents an innovative approach for alleviating continuous monocropping obstacles to improve P. heterophylla productivity and quality. • Trichoderma consortium compost corncob (TCCC) alleviates continuous monocropping obstacles of Pseudostellaria heterophylla. • TCCC promotes the growth and productivity and alleviates disease incidence of P. heterophylla under continuous monocropping condition. • TCCC enhances the soil nutrients and reshapes the soil microbial communities of continuous monocropping fields. • Enhancing soil nutrients and reshaping soil microbial communities are driving factors that promote plant growth and alleviate diseases. [ABSTRACT FROM AUTHOR]

In production of cereals like maize (Zea mays L.) and barley (Hordeum vulgare L.), seeds are often treated with pesticides and/or commercial products of plant beneficial microorganisms (PBM) to reduce possible root damage from insect pests and soil borne root diseases. In a field experiment with maize and barley under conservation agriculture, we examined how such seed treatments affected the resident root and soil microbiota. The seed treatments included a pesticide mixture and different commercial products of common PBM based on the biocontrol agents (BCA) Trichoderma harzianum and Metarhizium anisopliae alone and in combination and a mix of plant growth promoting rhizobacteria (PGPR), which were compared to a negative control without seed treatment. Soil and root samples were taken at two and three sampling times during the crop cycles for barley and maize, respectively, to measure root biomass, root colonization with mycorrhizal fungi and pathogens, soil microbial communities at a general taxonomic level using biomarker fatty acids, and ecological guilds of soil nematodes. Root health was monitored with observations of the presence of insect feeding larvae and root disease symptoms, which in general showed healthy roots during the full crop cycle. Overall, most of the root and soil biota variables measured changed during the crop cycle. However, for both crops, the seed treatments had no effects on the soil and root microbiota measured, except in the case of barley root infection with Polymyxa sp., which was reduced by all treatments. In conclusion, the pesticide and PBM seed treatments evaluated in the present study for maize and barley under conservation agriculture, in general, had limited effects on the resident root and soil microbiota. However, future studies should include complementary high-resolution sequencing methods when examining non-target effects of pesticides and microbial inoculants on the root and soil microbiota. [ABSTRACT FROM AUTHOR]

This study assessed the effectiveness of Trichoderma asperelloides SKRU-01 and Trichoderma asperellum NST-009 in controlling postharvest anthracnose in chili fruits caused by Colletotrichum gloeosporioides PSU-03, using both in vitro assays and studies on chili fruits. In vitro findings indicated that both SKRU-01 and NST-009 exhibited inhibitory effects on PSU-03 through synergistic mechanisms, involving competition for space and nutrients, as well as the production of non-volatile and volatile compounds. Culture filtrates of both strains, at low concentrations (diluted at 1/1000), displayed a robust antifungal effect on PSU-03, achieving over 50% inhibition. This efficacy, however, was lower than that achieved by five chemical fungicides (propiconazole®, prochloraz®, metalaxyl®, azoxystrobin®, and thiram®). The efficacy against anthracnose of SKRU-01 (60.34% protection) and NST-009 (62.07% protection) was significantly higher than metalaxyl®, azoxystrobin®, and thiram®, but lower than propiconazole® and prochloraz®. The two strains demonstrated both preventive and curative effects. Furthermore, results from tests on chili seed germination showed that culture filtrates SKRU-01 significantly inhibited the germination of chili pepper seeds and the growth of seedlings in lab settings. Overall, this study suggests that both SKRU-01 and NST-009 hold the potential for managing postharvest anthracnose in chili fruits caused by C. gloeosporioides. It is crucial to note, however, that culture filtrates SKRU-01 should not be used for biostimulating root development in chili peppers. This research provides valuable insights into the potential application of these biocontrol agents, contributing to enhancing the quality and shelf life of chili produce. • T. asperelloides SKRU-01 and T. asperellum NST-009 could control chili anthracnose. • Both strains competed and produced volatile organic compounds and non volatiles. • The two strains demonstrated both protective and curative effects. • SKRU-01 non-volatiles inhibited chili pepper seed germination and seedling growth. [ABSTRACT FROM AUTHOR]

Pine wilt disease caused by Bursaphelenchus xylophilus is an extremely serious issue worldwide. Trichoderma asperellum is used as a biocontrol agent due to its efficient inhibition of various pathogens. Three solvents, methanol, petroleum ether, and acetone, were used for extracting the fermentation broth of T. asperellum. All three extracts displayed a significant difference in nematocidal efficacy of more than 70% at 72 hours. Water-soluble natural products or polyphenolic compounds exhibited the greatest nematocidal ability after 24 hours of treatment with methanol. However, lipophilic substances exhibit long-lasting nematocidal ability. During this process, the antioxidant enzyme activities of T. asperellum significantly increased, including those of catalase, superoxide dismutase, and peroxidase. The interaction between T. asperellum and B. xylophilus induces the synthesis of glycoside hydrolase family 18 (chitinases) and peptidase S8, potential subtilisin, which dissolve the body wall of the nematodes. Fatty acid metabolism was significantly increased, and a defensive response against substances secreted by the nematode was produced. Antioxidant reaction mechanisms and detoxification metabolism were enriched and included a large number of upregulated DEGs, including GST , CYP450 , LysM , and RTA1. The dual and toxic mechanisms exhibited by T. asperellum during its response to B. xylophilus involve multiple pathways and genes that are collectively regulated. This is the first study to investigate the effects of T. asperellum on B. xylophilus. This study provides a theoretical basis for the potential use of T. asperellum as a biocontrol agent against B. xylophilus. [Display omitted] • All three extracts had a significant difference in nematicidal efficacy of more than 70%. • T. asperellum increased antioxidant enzyme activities including CAT, SOD, and POD. • Detoxification metabolism was enriched, including GST, CYP450, LysM, and RTA1. • Chitinases, and subtilisin dissolved the body wall of the nematodes. • In the network, three transporter and four oxidases in highly connected genes. [ABSTRACT FROM AUTHOR]

The use of growth-promoting microorganisms with biostimulant characteristics is an important biological asset for the acclimatization of micropropagated seedlings. The present study aimed to evaluate the efficacy of the application of Trichoderma spp. on the promotion of the growth of micropropagated banana seedlings during acclimatization. The experiment was performed in an 8 × 6 completely randomized design using the following treatments: water, seedlings fertilized with controlled-release fertilizer, commercial biological inputs (A: T. asperellum , B/C: T. harzianum), and LPPC299 and LPPC300 strains. Plant height, pseudostem diameter, number of leaves, total leaf area, root length, fresh and dry mass of the plant, and accumulation of sodium, macronutrients, and micronutrients were evaluated 60 days after inoculation. Strains LPPC299 and LPPC300 were subjected to molecular identification by DNA sequencing of the ITS/5.8S locus. In vitro detection of growth promotion-related mechanisms and mycelial growth of biostimulants were performed using scanning electron microscopy. LPPC299 and LPPC300 had a greater similarity to T. longibrachiatum. LPPC299 was able to promote greater pseudostem diameter, number of leaves, and total leaf area in banana seedlings. T. asperellum (A) favored seedling performance in terms of fresh and dry mass of the plants. The strains were able to produce siderophores, indoleacetic acid, and catalase in vitro. Seedlings inoculated with the strains accumulated Mn, S (LPPC300), and Mg (LPPC299). LPPC299 from the banana rhizosphere was efficient in promoting performance in banana seedlings, showing its potential as a biostimulant for this crop. [Display omitted] • Trichoderma longibrachiatum is associated with the rhizosphere of banana trees. • T. longibrachiatum produces siderophores, indoleacetic acid and catalase. • T. longibrachiatum increases the growth of banana seedlings under acclimatization. • T. longibrachiatum increases the build-up of nutrients in banana under acclimatization. [ABSTRACT FROM AUTHOR]

• Trichoderma is extensively used in agriculture as a biocontrol agent. • The article describes the role of proteins from Trichoderma in plant pathogen suppression as well as in the activation of plant immune response. Trichoderma spp, due to their biocontrol attributes have been explored in agriculture to combat the plant diseases. Since its exploration as a biocontrol agent, the mechanistic insights have shown the production of various lytic enzymes and secondary metabolites are responsible for its ability to suppress plant pathogens. Proteins such as lytic enzymes which include chitinases, glucanases, and proteases have been reported for targeting the cell wall and other components of the fungal plant pathogens have been characterized. More recently, with the advent of omics technologies, proteins of Trichoderma origin in the form of elicitors, histone modification and others have been identified for their role in boosting the plant immune responses through systemic acquired resistance (SAR) or induced systemic resistance (ISR). Here, we have described the role of extracellular proteins from Trichoderma origin and their role in pathogen suppression as well as activation of plant immune response. [ABSTRACT FROM AUTHOR]

Cornmint (Mentha arvensis) is cultivated to produce essential oil, which consists of about 75% of menthol. Fresh biomass is hydrodistilled for ∼5 h to obtain essential oil (1%), and the rest of the pre-treated biomass (99%) is generated as waste. A novel, green and economical two-step process has been developed using a mixture of imidazole (IM:1 M)-p-toluene sulfonic acid (pTSA:1.2 M), and IM (0.2 M)-20%NH 3 for the separation of lignin and hemicellulose, respectively. The lignin and hemicellulose were isolated from the respective solution by precipitation, and final undissolved solid residue was obtained as cellulose. This process was scaled-up, recovering cellulose (38%), hemicellulose (27%), and lignin (14%) using 7 L double jacketed reactor. The processing parameters such as temperature, solvent ratio, and time were optimized using single factorial design mathematical model for the isolation of biopolymers. Further, cellulose was enzymatically biotransformed to glucose through submerged and solid-state fermentation (SSF) using Trichoderma reesei, T. harzianum (TH, TH10) , and T. atroviride. Isolated cellulose was produced 61.5% of glucose at 30 °C, pH 5 in 72 h through SSF process using TH10 strain. This glucose solution was transformed to methyl levulinate (74%) under aqueous-methanol (5:1) solvent system for 2 h at 160 °C using La(OTf) 3.H 2 O catalyst. [Display omitted] [ABSTRACT FROM AUTHOR]

The biosynthesis of brasilane-type sesquiterpenoids (BTSs) attracts much attention owing to their unique skeleton of 5/6 bicyclic structure that contains five Me groups. Here, the crystal structures of a BTS cyclase TaTC6 from Trichoderma atroviride FKI-3849 and its complexes with farnesyl pyrophosphate (FPP) and analogue were reported. These structural information reveal that TaTC6 exploits a hydrophobic pocket to constrain the hydrocarbon region of FPP in a "U-shape" to facilitate the initial C1-C11 bond formation after pyrophosphate ionization. Following, four carbocations of reaction intermediates were molecularly docked into the hydrophobic pocket to reveal critical residues involved in the cyclization cascade. Finally, an S239-stabilized water molecule that is 3.9 Å away from the C8 of the last allyl cation may conduct hydration to quench the reaction cascade. Mutating S239 to alanine led to ca. 40% reduction in activity compared with the wild-type enzyme. The conservation of the residues that constitute the hydrophobic pocket is also discussed. Overall, this study will give an insight into the mechanism of how the active site of STCs constrain the conformation of the flexible FPP and series allylic carbocations for the complicated-ring formation and unusual carbon rearrangement in the biosynthesis of BTSs. [ABSTRACT FROM AUTHOR]

Pythium species are devasting pathogens causing major crop losses, e.g., damping-off in sugar beet caused by Pythium ultimum and root-rot of tomato caused by Pythium aphanidermatum. The use of natural antagonistic microorganisms is a promising environment-friendly approach to control Pythium -caused plant diseases. There are several examples of biocontrol of diseases caused by Pythium species but the application of bioeffectors (biological control agents) is limited for various reasons, including the restricted amount of gene-modification based biotechnological progress. The regulations in many countries prevent genetically modified bioeffectors from being routinely deployed in field conditions. Our two connected aims in this review are (1) to compile and assess achievements in genetic modification of bioeffectors which have been tested for parasitism or antagonism towards a Pythium plant pathogen or biocontrol of a plant disease caused by a Pythium species, and (2) discuss how a better performing bioeffector could be engineered to improve biocontrol of Pythium -caused plant diseases. We focus on the role of seven key mechanisms: cellulases, carbon catabolite de-repression, glycosylation, reactive oxygen species, chitin re-modelling, proteases, and toxic secondary metabolites. Genetic modifications of bioeffectors include gene deletion and overexpression, as well as the replacement of promoter elements to tune the gene expression to the presence of the pathogen. Gene-modifications are limited to fungal and bacterial bioeffectors due to the difficulty of gene modification in oomycete bioeffectors such as Pythium oligandrum. We assess how previous gene modifications could be combined and what other gene modification techniques could be introduced to make improved bioeffectors for Pythium -caused plant diseases. The broad host-range of Pythium spp. suggests engineering improved antagonistic traits of a bioeffector could be more effective than engineering plant-mediated traits i.e. , engineer a bioeffector to antagonise a plant pathogen in common with multiple plant hosts rather than prime each unique plant host. • Bioeffectors of Pythium -caused diseases reviewed in an ecological context. • Seven important mechanisms of biocontrol of Pythium -caused diseases reviewed. • Mechanisms include offensive e.g., cellulases and defensive e.g., wall remodeling. • Several potential routes to stack these mechanisms to engineer a super-bioeffector. [ABSTRACT FROM AUTHOR]

[Display omitted] • Fermentation methods partially determined transcriptional response of strains. • Trichoderma asperellum T-1 effectively deconstructed the structure of wheat straw. • Secretion of carbohydrate-binding modules enhanced straw deconstruction by T-1. • T. asperellum T-1 is suitable for both solid state and submerged fermentation. • T. reesei QM6a is preferred used in solid state fermentation. Microorganisms harvest energy from agricultural waste by degrading its structure. By comparing with Trichoderma reesei QM6a in cellulase production, straw deconstruction and transcriptome response, Trichoderma asperellum T-1 was identified to be prioritized for the fermentation of natural straw. Cellulase activity of T-1 was 50%–102% higher than QM6a. And the degradation rate of hemicellulose and ligin in wheat straw by T-1 reached 40% and 42%. Time-driven changes in the gene expression of extracellular proteins involved in polysaccharide, xylan, and hemicellulose metabolism and hydrolysis indicated that T-1 positively responded in both solid state fermentation and submerged fermentation for lignocellulose degradation. A significantly enriched category encoding carbohydrate-binding modules is considered critical for the deconstruction of the natural structure by T-1. The findings highlight the superiority of T. asperellum T-1 in straw fermentation, base on which, the construction of efficient microbial agents is expected to enhance the utilization of biomass. [ABSTRACT FROM AUTHOR]

In the face of agricultural challenges posed by both abiotic and biotic stressors, phytopathogens emerge as formidable threats to crop productivity. Conventional methods, involving the use of pesticides and microbes, often lead to unintended consequences. In addressing this issue, ICAR -Indian Institute of Oilseeds Research (ICAR-IIOR) has developed a chitosan-based double-layer seed coating. Emphasizing crop input compatibility, entrapment, and characterization, the study has yielded promising results. The double-layer coating on groundnut seeds enhanced germination and seedling vigor. Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) confirmed the structural changes and entrapment of crop inputs. The persistence of T. harzianum (Th4d) and Bradyrhizobium sp. in chitosan blended film in studied soils revealed that viable propogules of Th4d were recorded in double layer treatment combination with 3.54 and 3.50 Log CFUs/g of soil (colony forming units) and Bradyrhizobium sp. with 5.34 and 5.27 Log CFUs/g of soil at 90 days after application (DAA). Root colonization efficacy studies of Th4d and Bradyrhizobium sp. in groundnut crop in studied soils revealed that, maximum viable colonies were observed at 45 days after sowing (DAS). This comprehensive study highlights the potential of chitosan-based double-layer seed coating providing a promising and sustainable strategy for stress management in agriculture. [Display omitted] [ABSTRACT FROM AUTHOR]

• Two new steriods were identified and evaluated the cytotoxicity and antifungal activity. • Compound 1 was the first identified steroid containing γ-butyrolactone moiety at C-3 positon. Chemical investigation of the fungus Trichoderma asperellum SCNU-F0048 led to the discovery of two new steroids, ergosta-4,6,8 (14),22-tetraen-3-(3′-methyl-4′-hydroxyl- γ -butenolide) (1) and camphosterol B (2), as well as two known compounds, i.e. stigmasta-4,6,8(14),22-tetraen-3-one (3) and 4-hydroxy-17- methylincisterol (4). Their structures were elucidated by extensive nuclear mangnetic resonance, spectrum analysis and single crystal X-ray diffraction analysis. Bioassay disclosed that compound 1 showed strong cytotoxicity to a panel of tumor cell lines. Moreover, compounds 1 and 2 showed excellent antifungal activity against Penicillium italicum with IC 50 values of 0.016 and 0.022 μM, respectively. [ABSTRACT FROM AUTHOR]

In the understanding of the molecular interaction between plants and their microbiome, a key point is to identify simplified models of the microbiome including relevant bacterial and fungal partners which could also be effective in plant growth promotion. Here, as proof-of-concept, we aim to identify the possible molecular interactions between symbiotic nitrogen-fixing rhizobia and soil fungi (Trichoderma spp.), hence shed light on synergistic roles rhizospheric fungi could have in the biology of symbiotic nitrogen fixation bacteria. We selected 4 strains of the model rhizobium Sinorhizobium meliloti and 4 Trichoderma species (T. velutinum , T. tomentosum , T. gamsii and T. harzianum). In an experimental scheme of 4 ×4 strains x species combinations, we investigated the rhizobia physiological and transcriptomic responses elicited by fungal spent media, as well as spent media effects on rhizobia-host legume plant (alfalfa, Medicago sativa L.) symbiosis. Fungal spent media had large effects on rhizobia, specific for each fungal species and rhizobial strains combination, indicating a generalized rhizobia genotype x fungal genotype interaction, including synergistic, neutral and antagonistic effects on alfalfa symbiotic phenotypes. Differential expression of a high number of genes was shown in rhizobia strains with up to 25% of total genes differentially expressed upon treatment of cultures with fungal spent media. Percentages over total genes and type of genes differentially expressed changed according to both fungal species and rhizobial strain. To support the hypothesis of a relevant rhizobia genotype x fungal genotype interaction, a nested Likelihood Ratio Test indicated that the model considering the fungus-rhizobium interaction explained 23.4% of differentially expressed genes. Our results provide insights into molecular interactions involving nitrogen-fixing rhizobia and rhizospheric fungi, highlighting the panoply of genes and genotypic interactions (fungus, rhizobium, host plant) which may concur to plant symbiosis. • The plant symbiotic bacterium Sinorhizobium meliloti is affected by the presence of the fungus Trichoderma. • Trichoderma modulates a large fraction of Sinorhizobium meliloti genes. • 30% of transcriptomic changes are specific to S. meliloti and Trichoderma strains combinations. • Specific S. meliloti - Trichoderma strains combinations can produce synergistic effects on the quality of symbiosis. [ABSTRACT FROM AUTHOR]

Investigating the effects of microplastics remains a pertinent issue given the extensive biodiversity of exposed organisms, variations in the mechanisms of action among individual polymers, and their reliance on environmental conditions. Therefore, the objective of the study was to investigate some interactions of the popular biodegradable plastic poly(butylene adipate-co-terephthalate) (PBAT) on some aspects of the physiology of filamentous fungi Trichoderma spp., integral components of the soil microbiome known for promoting plant growth and aiding in pollution remediation. In the two Trichoderma strains examined, a positive influence on fungal growth was observed in the presence of microplastics (MPs). The capability to degrade MPs with the involvement of cytochrome P450 monooxygenases was confirmed through the detection of terephthalic acid (TPA) in postculture extractions. Additionally, inhibition of degradative activity in the presence of MPs was observed for metolachlor (MET) and 2,4-dichlorophenoxyacetic acid (2,4-D). Moreover, biodegradation pathways with the involvement several dechlorinated or hydroxylated metabolites were proposed for examined compunds. Concentration-dependent shifts in oxidative stress enzymes activity and slight modifications in phospholipid profiles were noted for the tested PBAT concentrations. In summary, this study enhances our understanding of PBAT's impact on the physiology of filamentous fungal and their degradative capacities important for mitigating environmental pollution by xenobiotics and plastics. [Display omitted] • The toxicity of PBAT microparticles is concentration-dependent. • Fungi show varying levels of sensitivity in different culturing conditions. • Trichoderma sp. is capable of degrading PBAT. • The presence of PBAT can impact the herbicide degradation process. • Trichoderma sp. is able to absorb microplastic into cells. [ABSTRACT FROM AUTHOR]

Endo-β-1,4-xylanases degrade heteroxylans that constitute the lignocellulosic plant cell wall. This enzyme is widely used in the food, paper, textile, and biorefinery industries. Temperature affects the optimum activity of xylanase and is an important factor in its application. Various structural analyses of xylanase have been performed, but its structural influence by temperature is not fully elucidated. To better understand the structural influence of xylanase due to temperature, the crystal structure of xylanase II from Trichoderma longibrachiatum (TloXynII) at room and cryogenic temperatures was determined at 2.1 and 1.9 Å resolution, respectively. The room-temperature structure of TloXynII (TloXynIIRT) showed a B-factor value 2.09 times higher than that of the cryogenic-temperature structure of TloXynII (TloXynIICryo). Subtle movement of the catalytic and substrate binding residues was observed between TloXynIIRT and TloXynIICryo. In TloXynIIRT, the thumb domain exhibited high flexibility, whereas in TloXynIICryo, the finger domain exhibited high flexibility. The substrate binding cleft of TloXynIIRT was narrower than that of TloXynIICryo, indicating a distinct finger domain conformation. Numerous water molecule networks were observed in the substrate binding cleft of TloXynIICryo, whereas only a few water molecules were observed in TloXynIIRT. These structural analyses expand our understanding of the temperature-dependent conformational changes in xylanase. [Display omitted] • The crystal structures of TloXynII at room and cryogenic temperatures were determined. • TloXynIIRT and TloXynIICryo exhibited different conformations for the catalytic and substrate-binding residues. • The substrate-binding cleft of TloXynIIRT was narrower than that of TloXynIICryo. • TloXynIIRT showed a B-factor value that was 2.09 times higher than that of TloXynIICryo. • The regions of higher flexibility differed between TloXynIIRT and TloXynIICryo. [ABSTRACT FROM AUTHOR]

The escalating global climate anomalies and disruption of ecological equilibrium pose profound threats to agroecosystems and crop productivity. Drought stress, a prominent consequence of climate change, exert detrimental effects on plant physiology, thereby reducing crop yield. This study explores the implementation of Trichoderma -mediated defence priming in barley under drought stress. Through a comprehensive evaluation of encompassing morphological, stomatal, biochemical, and yield parameters, Trichoderma - primed barley is compared with non-primed barley under drought stress conditions. Our results revealed that Trichoderma -primed barley exhibits heightened tolerance to drought stress in comparison to non-primed barley. The improved stomatal characteristics indicate that Trichoderma priming enables the plant to better adapt to drought conditions. Biochemical analysis showed enhanced photosynthetic pigments and reduced MDA in primed barley under drought, suggesting that primed barley experiences less oxidative stress compared to non-primed barley. Additionally, Trichoderma -primed plants displayed heightened levels of both non-enzymatic and enzymatic antioxidant activities, aiding in ROS detoxification. Stress responses are activated upon encountering a triggering stimulus (drought stress), but not after Trichoderma priming. This means there was no wasteful diversion of resources towards resistance mechanisms in the absence of stressful situations, leading to enhanced yield for primed barley compared to non-primed barley under stressful conditions. Furthermore, our investigation into the heritability of priming effects in the succeeding generation demonstrates that the progeny of primed barley retains enhanced protection akin to their primed parents. In contrast the progeny of non-primed barley exhibits greater vulnerability under water-deficit conditions. The expression of the epigenetic regulator gene HvDME was more enhanced in primed barley and its subsequent generations than in non-primed barley. This research emphasizes the potential of Trichoderma priming as a cost-effective strategy to enhance barley production amidst climate change challenges. The priming defense approach offers an intelligent plant health management solution devoid of adverse environmental or yield-related implications. This discovery holds significant promise, particularly in regions with limited water resources where crops are highly susceptible to drought stress. The findings contribute to the advancement of sustainable and climate-resilient agriculture, offering insights for practitioners and policymakers aiming to address the adverse impacts of climate change on crop production. • Trichoderma priming significantly enhances growth parameters and drought tolerance in barley. • Trichoderma- mediated defense priming prevents wasteful consumption of metabolites under stress-free conditions. • Primed barley exhibits heightened yield-related traits under water stress compared to non-primed barley. • Trichoderma priming not only persists throughout the lifespan of the barley but is also inherited by the next generation. • The progeny of primed barley demonstrates superior performance compared to that of non-primed barley under drought stress. [ABSTRACT FROM AUTHOR]

The stigmas of the medicinal plant Crocus sativus are utilized as an expensive traditional Chinese medicine to treat irregular menstruation, depression, postpartum thrombosis, and bruises. As one of the most severe diseases of C. sativus , corm rot has hindered development of the C. sativus industry. Synthetic microbial communities (SMCs) have shown significant promise for the control of plant diseases, but there are few reports on their use for control of C. sativus corm rot. In this study, we co-cultured pairs of C. sativus rhizosphere soil-isolated fungi and bacteria, performed compatibility experiments on them to obtain SMCs with good compatibility, and evaluated their ability to improve disease resistance against C. sativus corm rot induced by Fusarium oxysporum. The fermentation broth of the most effective SMC was partitioned into five fractions, and disease resistance activity assays were conducted to identify the fraction in which active metabolites were concentrated. Metabolomic analyses were carried out to analyze the chemical composition of this fraction and determine how it differed from that of single cultures. We constructed four SMCs and found that SMC03 (Trichoderma yunnanense SR38 + Paenibacillus peoriae SR235) showed the best disease resistance activity, which was significantly higher than that of SR38 or SR235 alone (P < 0.05). The active metabolites produced by SMC03 were mainly concentrated in the aqueous fraction, which contained abundant organic acids and their derivatives. Co-cultivation of SR38 and SR235 increased overall production of DL-3-phenyllactic acid, 3-hydroxydecanoic acid, and (2S)-2-isopropylmalate, and we speculate that these components can bind membrane receptors of plant cells, induce local resistance, and subsequently generate plant immune response, thereby enhancing C. sativus disease resistance. In summary, co-cultivation of a fungus and a bacterium from the C. sativus rhizosphere may enhance host resistance to corm rot by accelerating the synthesis of certain organic acids. This study demonstrates for the first time the feasibility of developing SMCs for control of corm rot in C. sativus , which is of great significance in guiding the development of efficient biocontrol agents against C. sativus corm rot, thereby promoting sustainable cultivation of C. sativus. These findings also provide reference for the biocontrol of fungal diseases in other field cultivated medicinal plants. [Display omitted] • Corm rot has hindered development of Crocus sativus industry. • Co-culture of rhizosphere microorganisms may enhance plant disease resistance. • SMC03 (T. yunnanense + P. peoriae) showed the best biocontrol activity. • Certain organic acids may be associated with the higher disease resistance activity. [ABSTRACT FROM AUTHOR]

In the pursuit of sustainable turfgrass management for golf courses, a series of experiments was conducted to assess the potential of Trichoderma TrB (CNCM strain I-5327) as a natural solution. The research encompassed greenhouse and field trials across two golf courses. The comprehensive pot experiment investigated Trichoderma effectiveness, including native (TrB) and commercial strains, with and without organic amino acids, for turfgrass and soil health. The study followed a two-stage process, stimulating beneficial microorganisms with TrB and introducing Fusarium for biocontrol. Preliminary field trial on one golf course utilized a randomized block design to examine the effects of TrB and Trianum with amino acids, and fungicide on soil microbial community. A second field trial analyzed soil metabolic profiles after applying TrB, Trianum, and fungicide on another golf course. The greenhouse experiments demonstrated promising outcomes from the application of TrB, especially when combined with organic amino acids. This combination not only promoted plant growth and improved soil health but also effectively prevented the activation of Fusarium. In both field trials, it was observed that the introduction of TrB into the soil led to an increase in the population of soil fungi and bacteria and stimulated their activity. Our field data revealed that enriching the soil with TrB had a positive effect on soil microbial communities, while the application of fungicide resulted in a decrease in microbial activities. In summary, our research underscores Trichoderma 's potential (TrB) in sustainable golf course management. These findings highlight TrB as a promising natural solution for improving turfgrass health and soil quality in sustainable management of golf course. • TrB strain enhanced soil respiration via organic amino acids, promoting microbial and root activity. • TrB strain boosted NBI by 37.1 % during biocontrol, improving nutrient use and turfgrass health. • TrB strain surpassed Trianum in suppressing Fusarium, highlighting its biocontrol potential. • TrB strain impacted soil microorganism metabolism, restraining Fusarium for healthier soil. • TrB strain stimulated microbial activity, fostering nutrient cycling in golf fields, unlike fungicides. [ABSTRACT FROM AUTHOR]

Trichoderma spp. can enhance plant resistance against a wide range of biotic stressors. However, the fundamental mechanisms by which Trichoderma enhances plant resistance against Meloidogyne incognita , known as root-knot nematodes (RKNs), are still unclear. Here, we identified a strain of Trichoderma asperellum (T141) that could effectively suppress RKN infestation in tomato (Solanum lycopersicum L.). Nematode infestation led to an increase in the concentrations of reactive oxygen species (ROS) and malondialdehyde (MDA) in roots but pre-inoculation with T141 significantly decreased oxidative stress. The reduction in ROS and MDA was accompanied by an increase in the activity of antioxidant enzymes and the accumulation of flavonoids and phenols. Moreover, split root test-based analysis showed that T141 inoculation in local roots before RKN inoculation increased the concentration of phytohormone jasmonate (JA) and the transcripts of JA synthesis and signaling-related genes in distant roots. UPLC-MS/MS-based metabolomics analysis identified 1051 differentially accumulated metabolites (DAMs) across 4 pairwise comparisons in root division test, including 81 flavonoids. Notably, 180 DAMs were found in comparison between RKN and T141-RKN, whereas KEGG annotation and enrichment analysis showed that the secondary metabolic pathways, especially the flavonoid biosynthesis, played a key role in the T141-induced systemic resistance to RKNs. The role of up-regulated flavonoids in RKN mortality was further verified by in vitro experiments with the exogenous treatment of kaempferol, hesperidin and rutin on J2-stage RKNs. Our results revealed a critical mechanism by which T141 induced resistance of tomato plants against the RKNs by systemically promoting secondary metabolism in distant roots. • Trichoderma asperellum (T141) could effectively suppress RKN infestation in tomato. • T141 increased antioxidant enzyme activity and decreased RKN-induced oxidative stress. • T141 inoculation in local roots increased JA synthesis and signaling in distant roots. • Metabolomics study revealed a role for flavonoids in T141-induced systemic resistance. • T141 enhanced RKN resistance by systemically promoting secondary metabolism in tomato. [ABSTRACT FROM AUTHOR]

Enzymatic degradation of lignocellulosic biomass provides an eco-friendly approach to produce value-added macromolecules, e.g. , bioactive polysaccharides. A novel acidophilic GH5 β-1,4-endoglucanase (termed TaCel5) from Trichoderma asperellum ND-1 was efficiently expressed in Komagataella phaffii (∼1.5-fold increase, 38.42 U/mL). TaCel5 displayed both endoglucanase (486.3 U/mg) and alginate lyase (359.5 U/mg) enzyme activities. It had optimal pH 3.0 and strong pH stability (exceed 86 % activity retained over pH range 3.0–5.0). 80 % activity (both endoglucanase and alginate lyase) was retained in the presence of 15 % ethanol or 3.42 M NaCl. Analysis of action mode revealed that hydrolytic activity of TaCel5 required at least three glucose (cellotriose) residues, yielding mainly cellobiose. Glu241 and Glu352 are essential catalytic residues, while Asp106, Asp277 and Asp317 play auxiliary roles in cellulose degradation. TaCel5 displayed high hydrolysis efficiency for glucan and alginate substrates. ESI-MS analysis indicated that the enzymatic hydrolysates of alginate mainly contained disaccharides and heptasaccharides. This is the first detailed report of a bifunctional GH5 endoglucanase/alginate lyase enzyme from T. asperellum. Thus TaCel5 has strong potential in food and feed industries as a catalyst for bioconversion of cellulose- and alginate-containing waste materials into value-added products oligosaccharides, which was of great benefit both for the economy and environment. • A alginate-degrading endoglucanase (TaCel5) from T. asperellum was firstly studied. • It hydrolyzed glucan and cello-oligosaccharide to yield cellobiose. • No endoglucanase or alginate lyase activity was detected for mutants E241A and E352A. • Enzymatic degradation of alginate mainly yielded disaccharides and heptasaccharides. • TaCel5 has great potential to produce biopolysaccharides from agricultural waste. [ABSTRACT FROM AUTHOR]

Trichoderma can enhance the metabolism of organophosphate pesticides in plants, but the mechanism is unclear. Here, we performed high-throughput transcriptome sequencing of roots upon Trichoderma asperellum (TM) inoculation and phoxim (P) application in tomato (Solanum lycopersicum L.). A total of 4059 differentially expressed genes (DEGs) were obtained, including 2110 up-regulated and 1949 down-regulated DEGs in P vs TM+P. COG and KOG analysis indicated that DEGs were mainly enriched in signal transduction mechanisms. We then focused on the pesticide detoxification pathway and screened out cytochrome P450 CYP736A12 as a putative gene for functional analysis. We suppressed the expression of CYP736A12 in tomato plants by virus-induced gene silencing and analyzed tissue-specific phoxim residues, oxidative stress markers, glutathione pool, GST activity and related gene expression. Silencing CYP736A12 significantly increased phoxim residue and induced oxidative stress in tomato plants, by attenuating the TM-induced increased activity of antioxidant and detoxification enzymes, redox homeostasis and transcripts of detoxification genes including CYP724B2 , GSH1 , GSH2 , GR , GPX , GST1 , GST2 , GST3 , and ABC. The study revealed a critical mechanism by which TM promotes the metabolism of phoxim in tomato roots, which can be useful for further understanding the Trichoderma -induced xenobiotic detoxification and improving food safety. [Display omitted] • Trichoderma asperellum (TM) reduced phoxim (P) residue in tomato plants. • CYP736A12 detoxifying P in tomato was screened out as a putative gene by RNA-seq. • Silencing CYP736A12 increased phoxim residue and induced oxidative stress. • TM enhanced antioxidants, redox homeostasis and detoxification genes via CYP736A12. • CYP736A12 is essential for TM-induced phoxim metabolism in tomato roots. [ABSTRACT FROM AUTHOR]

Chemical investigation of deep-sea-derived fungus Trichoderma simmonsii ZEN3 led to the isolation of 16 compounds, including four steroids (1 − 4), four alkaloids (5 − 8), four polyketides (9 − 12), two lipids (13 and 14), one oxygenated cyclohexane derivative (15), and one harziandione (16). The structures of these compounds were determined mainly by detailed analysis of NMR spectra data. All compounds were isolated from T. simmonsii for the first time. Compounds 3 − 5 , 7 , 8 , and 15 were isolated from the genus Trichoderma for the first time, and could be chemotaxonomic markers that distinguished T. simmonsii from other Trichoderma species. Compounds 11 , 13 , and 16 could be chemotaxonomic markers of the genus Trichoderma. [Display omitted] • The deep-sea-derived fungus Trichoderma simmonsii has been investigated. • Sixteen miscellaneous compounds have been isolated. • Compounds 11 , 13 and 16 are considerable Trichoderma genus chemotaxonomic markers. • T. simmonsii chemotaxonomically distinguishes based on compounds 3 − 5 , 7 , 8 , and 15. [ABSTRACT FROM AUTHOR]

Drought-induced stress represents a significant challenge to agricultural production, exerting adverse effects on both plant growth and overall productivity. Therefore, the exploration of innovative long-term approaches for addressing drought stress within agriculture constitutes a crucial objective, given its vital role in enhancing food security. This article explores the potential use of Trichoderma , a well-known genus of plant growth-promoting fungi, to enhance plant tolerance to drought stress. Trichoderma species have shown remarkable potential for enhancing plant growth, inducing systemic resistance, and ameliorating the adverse impacts of drought stress on plants through the modulation of morphological, physiological, biochemical, and molecular characteristics. In conclusion, the exploitation of Trichoderma 's potential as a sustainable solution to enhance plant drought tolerance is a promising avenue for addressing the challenges posed by the changing climate. The manifold advantages of Trichoderma in promoting plant growth and alleviating the effects of drought stress underscore their pivotal role in fostering sustainable agricultural practices and enhancing food security. [ABSTRACT FROM AUTHOR]

In many eukaryotes, small RNAs (sRNAs) can mediate gene expression regulation through a mechanism known as RNA silencing. In fungi, RNA silencing plays a crucial role in numerous biological processes, including parasitic and mutualistic fungus-plant interactions. This review summarizes recent findings on the role of RNA silencing in parasitic fungus-fungus and fungus-insect interactions in relation to their use for the biological control (biocontrol) of fungal plant diseases and insect damage. Genes belonging to the RNA silencing machinery are identified in the genomes of almost all known fungal and oomycete biocontrol organisms. However, recent functional genetic studies in Ascomycota species of the Hypocreales order, such as Trichoderma atroviride and Clonostachys rosea , show how RNA silencing can have family-specific effects, as conidiation is affected differently in the two organisms when the same elements of the RNA silencing machinery are deleted. The size of sRNAs regulated by RNA silencing can also vary between organisms. Cross-species RNA silencing represents a new field in the study of antagonistic interactions. For example, a microRNA (miRNA) of another hypocrealean fungus, Beauveria bassiana, was proven to target genes involved in the immune response of mosquitoes, and there are indications that miRNAs from the mycoparasitic C. rosea may target factors of virulence in its plant-pathogenic host fungi. Accumulating evidence from many species shows that the number of endogenous genes affected by the disruption of the RNA silencing mechanism is always much higher than the number of predicted direct target genes. As several putative targets of fungal sRNAs are transcription factors, it is possible that specific sRNAs have a role as master regulators of gene expression, affecting the transcription of a high number of genes through cascading regulating effects. The challenges faced when studying cross-species RNA silencing, including sRNA trafficking during mycoparasitism, are also discussed. This includes the difficulties in separating the extracellular vesicles of mycoparasitic fungi from those of their hosts, the high amount of sequencing reads lost in bioinformatics filtering steps, imprecise target prediction and the lack of a streamlined accepted way of reporting results. • Genes encoding proteins involved in the production of small RNAs are present in most fungi, including biocontrol agents. • Disruption of the RNA silencing machinery has major effects on the phenotype and effectiveness of biocontrol agents. • Many small RNAs are constitutively produced but downregulated upon interaction with hosts. • Some small RNAs can be translocated to the host and have an effect, thereby hijacking the host's RNA silencing machinery. • Some sRNAs are master regulators, able to affect the expression of many genes by targeting transcription factors. [ABSTRACT FROM AUTHOR]

Species of fungal genus Trichoderma are characterized by a versatile lifestyle, high adaptability to the changing environmental conditions and the ability to establish sophisticated interactions with other organisms. Due to their ability to antagonize plant pathogens and to elicit the plant defence responses against biotic/abiotic stresses, Trichoderma spp. are commonly used as commercially biopesticides and biofertilizers. The Trichoderma success in the rhizosphere is supported by a wide arsenal of specialised metabolites (SMs) providing morphological and physiological autoregulation, self-protection and facilitating fungal communication. This review aims to explore the roles of SMs in the biology of fungi, with special emphasis on the genus Trichoderma and on how divergence in the SMs genetic structure determine Trichoderma lifestyles. Trichoderma genomes are endowed with a high number of SMs biosynthetic genes, and understanding the genetic basis of their biosynthesis is crucial for determining the role of these metabolites in Trichoderma ecophysiology and for expanding their application in crop protection. Recent advances on the characterization of the Trichoderma SMs genetic inventory driven by computational biology are discussed. [Display omitted] • Secondary metabolites are messengers in fungal communication and stress tolerance • The diversity of Trichoderma lifestyles is supported by their secondary metabolites • Broad-function bioactive molecules support Trichoderma success in the rhizosphere • Ecologic divergences influence the genetic structure of Trichoderma metabolomes [ABSTRACT FROM AUTHOR]

Trichoderma spp. are widely used as plant disease biocontrol agents in agriculture. Mycoparasitism, which is an ancestral trait of Trichoderma , is one of the most important mechanisms of reducing the pathogen inocula. Mycoparasitism is a complex physiological process that should be viewed in the broad perspective of microbial competition, and involves the production of enzymes and secondary metabolites. Trichoderma spp. have traditionally been viewed as necrotrophic mycoparasites; however, there are evidences that, at least in some instances, they behave as hemibiotrophs, causing minor damage to the host cell wall and having an intracellular existence in the host cell for a significant period. In this review, we cover different aspects of Trichoderma as mycoparasites, ranging from evolution to genomics and interactions with "non-target" fungi. • Several mycoparasitic Trichoderma species are used as plant disease biocontrol agents. • Some Trichoderma -fungal interactions could be hemibiotrophic. • Mycoparasitism is programmed, from host sensing to invasion and finally death of host fungi. • Some strains are mycoparasitic on beneficial fungi like mycorrhizae and cultivated mushrooms. [ABSTRACT FROM AUTHOR]

• Spores of Trichoderma stromaticum have immunosuppressive effect and inhibit NETosis. • Exposure of neutrophil to spore reduces miRNAs such as miR-221, miR-222, miR-223, miR-23a and miR-27a. • Exposure of neutrophil to spore also reduces levels of expression of genes related to the formation of NETs. • Spores reduce the expression of neutrophil genes involved in exacerbated inflammatory processes. Although the genus Trichoderma is widely used as a biocontrol agent in crops, little is known about its potential impact on the human immune system. In mice, our group has shown that exposition to T. asperelloides spores lead to reduced neutrophil counts in the peripheral blood and in the peritoneal cavity. In addition, T. stromaticum spores produced an inflammatory infiltrate on mice lungs, reducing the levels of IFN-γ and IL-10 cytokines, reactive oxygen species, and receptors of microbial patterns. Here we demonstrate that the interaction of human peripheral neutrophils with T. stromaticum spores also leads to a reduced release of neutrophil extracellular traps (NETs) after induction with the NET-inducer agent phorbol 12-myristate 13-acetate. This interaction also reduced the expression levels of multiple microRNAs, such as miR-221, miR-222, miR-223 and miR-27a, as well as genes related to NETs, such as ELANE, MPO and PADI4. Furthermore, T. stromaticum spores affected the expression of the genes SOCS3, TLR4, CSNK2A1 , GSDMD, and NFFKBIA , related to the activation of inflammatory immune responses in neutrophils. Overall, our results suggest T. stromaticum as a potential NET inhibitor and as an immunomodulatory agent. Since this fungus is used as biocontrol in crops, our findings point to the importance of advancing our knowledge on the effects of this bioagent on the human immune system. Finally, the study of the active compounds produced by the fungus is also important for the prospection of new drugs that could be used to block the exacerbation of inflammatory immune responses present in several human diseases. [ABSTRACT FROM AUTHOR]

[Display omitted] • Two Trichoderma strains were isolated from the rhizospheric soil of medicinal plants. • Fermentation factors of lignocellulosic residues by Trichoderma strains was assessed. • Substrate components was a crucial factor affecting fermentation results. • T. afroharzianum showed pronounced cellulose degradation efficiency for corn straw. • Two Trichoderma strains would be useful for application in waste materials treatment. To achieve an economically viable substitutive strategy for lignocellulosic residues use, cellulose degradation by efficient Trichoderma strains become a key alternative. This study aimed to explore the effects of various cultural conditions on the degradation of different lignocellulosic substrates (corn straw and licorice residues) by Trichoderma longibrachiatum and Trichoderma afroharzianum in solid state fermentation. The strains were isolated from the rhizospheric soils of medicinal plants. In addition, the best fermentation combination was selected to evaluate the degradation results. This study confirmed that fermentation conditions and different components of corn straw and licorice residues were both crucial factors affecting the cellulase activity and spore production of two Trichoderma strains. A combination of T. afroharzianum degrading corn straw was selected for Response Surface Methodology optimization. The best fermentation conditions were fermentation time of 5.72 days, initial material liquid ratio of 1:4.58, initial pH of 6.32 and fermentation temperature of 28.8 ℃, with a filter paper cellulase activity of 0.91 U/mL. After continuous fermentation, the spore production reached 6.18 × 109 spores/g, the crude fiber degradation rate reached 23.5 % with the reducing sugar content maintained at 11 mg/g. These results highlight the great potential of T. afroharzianum degrading corn straw for organic amendment. [ABSTRACT FROM AUTHOR]

The use of biocontrol agents to control soilborne diseases is a promising alternative to chemical pesticides, however, obtaining a homogeneous distribution and incorporation of conidia of fungal biocontrol agents into the soil is often difficult. Several carriers/formulations have been proposed over time, unfortunately without offering an ultimate solution. We propose the use of wood pellets as a carrier of conidia of a saprophytic fungus that has good biodegradation and biocontrol properties (Trichoderma atroviride SC1). The coating process is based on the direct spraying of wood pellets with a conidial suspension at different rates. Beech, fir, and chestnut wood pellets were compared in terms of relevant physicochemical traits and efficacy in supporting the growth of the fungus. Beech wood pellets displayed the best characteristics in terms of water holding capacity, swelling properties, and disintegration time. T. atroviride SC1 grows best on beech and fir wood pellets and reaches a plateau after nine days of incubation, regardless of the initial coating concentrations. The addition of small quantities of a nitrogen source as tryptone or soy flour, soy proteins, and a mixture of animal proteins used as pet food to the conidial suspension can increase the growth by ten-folds on all types of wood pellets. Our results demonstrate that beech and fir wood pellets could be suitable carriers to deliver and sustain the growth of T. atroviride SC1. • New method for coating wood pellets with a conidial suspension of Trichoderma spp. • Beech wood showed the best physicochemical characteristics. • Trichoderma atroviride SC1 can grow on soft wood (fir) as well as hardwood (beech). • Adding tryptone (1 g/L) can enhance the growth of T. atroviride SC1 on wood. • Tryptone can be substituted by soy protein isolates for practical applications. [ABSTRACT FROM AUTHOR]

Trichoderma virens colonizes roots and develops a symbiotic relationship with plants where the fungal partner derives nutrients from plants and offers defence, in return. Tsp1, a small secreted cysteine-rich protein, was earlier found to be upregulated in co-cultivation of T. virens with maize roots. Tsp1 is well conserved in Ascomycota division of fungi, but none of its homologs have been studied yet. We have expressed and purified recombinant Tsp1, and resolved its structure to 1.25 Å resolutions, from two crystal forms, using Se-SAD methods. The Tsp1 adopts a β barrel fold and forms dimer in structure as well as in solution form. DALI based structure analysis revealed the structure similarity with two known fungal effector proteins: Alt a1 and PevD1. Structure and evolutionary analysis suggested that Tsp1 belongs to a novel effector protein family. Tsp1 acted as an inducer of salicylic acid mediated susceptibility in plants, rendering maize plants more susceptible to a necrotrophic pathogen Cochliobolus heterostrophus , as observed using plant defence assay and RT-qPCR analysis. • High resolution crystal structure reveals that Tsp1 adopts β-barrel fold. • Structure similarity of Tsp1 was predicted with two known fungal effector proteins: Alt a1 and PevD1. • RT-qPCR analysis and plant defence assay show that Tsp1 is an inducer of salicylic acid pathway in plants. [ABSTRACT FROM AUTHOR]

It is generally accepted that conidia, propagules of filamentous fungi, exist in the state of dormancy. This state is defined mostly phenomenologically, e.g., by germination requirements. Its molecular characteristics are scarce and are concentrated on the water or osmolyte content, and/or respiration. However, a question of whether conidia are metabolic or ametabolic forms of life cannot be answered on the basis of available experimental data. In other words, are mature conidia open thermodynamic systems as are mycelia, or do they become closed upon the transition to the dormant state? In this article, we present observations which may help to define the transition of freshly formed conidia to the putative dormant forms using measurements of selected enzyme activities, 1H- and 13C-NMR and LC-MS-metabolomes, and 14C-bicarbonate or 45Ca2+ inward transport. We have found that Trichoderma atroviride and Aspergillus niger conidia arrest the 45Ca2+ uptake during the development stopping thereby the cyclic (i.e., bidirectional) Ca2+ flow existing in vegetative mycelia and conidia of T. atroviride across the cytoplasmic membrane. Furthermore, we have found that the activity of α-ketoglutarate dehydrogenase was rendered completely inactive after 3 weeks from the conidia formation unlike of other central carbon metabolism enzymes. This may explain the loss of conidial respiration. Finally, we found that conidia take up the H14CO 3 - and convert it into few stable compounds within 80 d of maturation, with minor quantitative differences in the extent of this process. The uptake of H13CO 3 - confirmed these observation and demonstrated the incorporation of H13CO 3 - even in the absence of exogenous substrates. These results suggest that T. atroviride conidia remain metabolically active during first ten weeks of maturation. Under these circumstances, their metabolism displays features similar to those of chemoautotrophic microorganisms. However, the Ca2+ homeostasis changed from the open to the closed thermodynamic state during the early period of conidial maturation. These results may be helpful for studying the conidial ageing and/or maturation, and for defining the conidial dormant state in biochemical terms. [ABSTRACT FROM AUTHOR]

In fungi, MYB transcription factors (TFs) mainly regulate growth, development, and resistance to stress. However, as major disease-resistance TFs, they have rarely been studied in biocontrol fungi. In this study, MYB36 of Trichoderma asperellum Tas653 (Ta) was shown to respond strongly to the stress caused by Alternaria alternata Aa1004. Compared with wild-type Ta (Ta-Wt), the inhibition rate of the MYB36 knockout strain (Ta-Kn) on Aa1004 decreased by 11.06%; the superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities decreased by 82.15 U/g, 0.19 OD470/min/g, and 1631.2 μmol/min/g, respectively. The MYB36 overexpression strain (Ta-Oe) not only enhanced hyperparasitism on Aa1004, caused its hyphae to swell, deform, or even rupture, but also reduced the incidence rate of poplar leaf blight. MYB36 regulates downstream (TFs, detoxification genes, defense genes, and other antifungal-related genes by binding to the cis-acting elements "ACAT" and "ATCG". Zinc finger TFs, as the main antifungal TFs, account for 90% of the total TFs, and Zn37.5 (23.24-) and Zn83.7 (23.18-fold) showed the greatest expression difference when regulated directly by MYB36. The detoxification genes mainly comprised 11 major major facilitator superfamily (MFS) genes, among which MYB36 directly increased the expression levels of three genes by more than 2−3.44-fold. The defense genes mainly encoded cytochrome P450 (P450) and hydrolases. e.g., P45061.3 (2−10.95-), P45060.2 (2−7.07-), and Hyd44.6 (2−2.30-fold). This study revealed the molecular mechanism of MYB36 regulation of the resistance of T. asperellum to A. alternata and provides theoretical guidance for the biocontrol of poplar leaf blight and the anti-disease mechanism of biocontrol fungi. [ABSTRACT FROM AUTHOR]

The natural interactions between various bacteria, fungi, and other cellulolytic microorganisms destroy lignocellulosic polymers. The efficacy of this process is determined by the combined action of three main enzymes: endoglucanases, exo -glucanases, and β-glucosidase. The enzyme attacks the polymeric structure's β-1,4-linkages during the cellulose breakdown reaction. This mechanism is crucial for the environment as it recycles cellulose in the biosphere. However, there are problems with enzymatic cellulose breakdown, including complex cellulase structure, insufficient degradation efficacy, high production costs, and post-translational alterations, many of which are closely related to certain unidentified cellulase properties. These issues impede the practical use of cellulases. A developing area of research is the application of this similar paradigm for industrial objectives. Cellulase enzyme exhibits greater promise in many critical industries, including biofuel manufacture, textile smoothing and finishing, paper and pulp manufacturing, and farming. However, the study on cellulolytic enzymes must move forward in various directions, including increasing the activity of cellulase as well as designing peptides to give biocatalysts their desired attributes. This manuscript includes an overview of current research on different sources of cellulases, their production, and biochemical characterization. [ABSTRACT FROM AUTHOR]

An undescribed trichodenone derivative (1), two new diketopiperazines (3 and 4) along with a bisabolane analog (2) were isolated from Trichoderma hamatum b-3. The structures of the new findings were established through comprehensive analyses of spectral evidences in HRESIMS, 1D and 2D NMR, Marfey's analysis as well as comparisons of ECD. The absolute configuration of 2 was unambiguously confirmed by NMR, ECD calculation and Mo 2 (AcO) 4 induced circular dichroism. Compounds 1 – 4 were tested for their fungicidal effects against eight crop pathogenic fungi, among which 1 showed 51% inhibition against Sclerotinia sclerotiorum at a concentration of 50 μg/mL. [Display omitted] • Three previously undescribed compounds were isolated form Trichoderma hamatum b-3. • The absolute configurations of all the isolated compounds were elucidated. • 1 showed 51% inhibition against Sclerotinia sclerotiorum at a concentration of 50 μg/mL. [ABSTRACT FROM AUTHOR]

Trichoderma spp. are free-living fungi present in virtually all terrestrial ecosystems. These soil fungi can stimulate plant growth and increase plant nutrient acquisition of macro- and micronutrients and water uptake. Generally, plant growth promotion by Trichoderma is a consequence of the activity of potent fungal signaling metabolites diffused in soil with hormone-like activity, including indolic compounds as indole-3-acetic acid (IAA) produced at concentrations ranging from 14 to 234 μg l-1, and volatile organic compounds such as sesquiterpene isoprenoids (C 15), 6-pentyl-2 H -pyran-2-one (6-PP) and ethylene (ET) produced at levels from 10 to 120 ng over a period of six days, which in turn, might impact plant endogenous signaling mechanisms orchestrated by plant hormones. Plant growth stimulation occurs without the need of physical contact between both organisms and/or during root colonization. When associated with plants Trichoderma may cause significant biochemical changes in plant content of carbohydrates, amino acids, organic acids and lipids, as detected in Arabidopsis thaliana , maize (Zea mays), tomato (Lycopersicon esculentum) and barley (Hordeum vulgare), which may improve the plant health status during the complete life cycle. Trichoderma -induced plant beneficial effects such as mechanisms of defense and growth are likely to be inherited to the next generations. Depending on the environmental conditions perceived by the fungus during its interaction with plants, Trichoderma can reprogram and/or activate molecular mechanisms commonly modulated by IAA, ET and abscisic acid (ABA) to induce an adaptative physiological response to abiotic stress, including drought, salinity, or environmental pollution. This review, provides a state of the art overview focused on the canonical mechanisms of these beneficial fungi involved in plant growth promotion traits under different environmental scenarios and shows new insights on Trichoderma metabolites from different chemical classes that can modulate specific plant growth aspects. Also, we suggest new research directions on Trichoderma spp. and their secondary metabolites with biological activity on plant growth. • The fungal genus Trichoderma fulfills multiple ecological functions in the rhizosphere. • Several Trichoderma species can promote plant growth in natural and agricultural systems. • Trichoderma spp. produce a large number of bioactive metabolites that exert beneficial effects on plant growth. • Different species of these fungi can solubilize soil nutrients. • Trichoderma can promote the adaptation of plants under stressful conditions. [ABSTRACT FROM AUTHOR]

The scientific significance of this work involved in investigating the removal of contaminants of emerging concern (CECs) and antibiotic resistance genes (ARGs) at pilot-scale with partially saturated vertical flow constructed wetland either bioaugmented or not with the Trichoderma asperellum (strain T34) by means of the non-target liquid chromatography high-resolution mass spectrometry and SmartChip™ Real-Time PCR methodological approaches. Bioaugmentation testing was successful as Trichoderma was able to grow in competitive conditions resulting from the use of secondary treated domestic wastewater. There was a clear relationship between the removal efficiency (RE) and the biodegradability as well as the sorption capacity of the identified compounds. The most easily eliminated compounds were those which are known to be readily biodegradable (e.g., nicotine). Interestingly, from 22 compounds with RE > 65 %, 17 compounds have a log Dow between 0 and 3 (optimal plant uptake). The most significant beneficial effect was observed with benzotriazole and diclofenac. The most difficult compounds to be eliminated were the most polar ones (log Dow < 0) such as melamine. High variability in removal efficiency over the course of a four-week experiment remained the chief constraint for the proper assessment of treatment performances. Transformation products (TPs) were also investigated. N-oxide TPs and 14-hydroxyclarithromycin were formed in wetland treatment while TPs which were prone to undergo further N-dealkylation or hydroxylation reactions were partly eliminated. Bioaugmented wetland treatments caused a shift in the composition of ARGs but there was no statistical difference in average rates of ARGs removal. [Display omitted] • Bioaugmented wetland with Trichoderma was evaluated by non-target LC-HRMS screening. • Trichoderma was able to grow in real competitive conditions. • Removal efficiency higher than 65 % was observed for slow biodegradable contaminants. • N-oxide TPs and 14-hydroxyclarithromycin were formed in treatment wetland. • Treatment wetland failed to reduce the overall bacterial and ARGs loads. [ABSTRACT FROM AUTHOR]

Microbial co-culture has proven to be an effective way to improve the ability of microorganisms to biocontrol. However, the interactive mechanisms of co-cultural microbes, especially between fungi and bacteria, have rarely been studied. By comparative analysis of morphology, transcriptomics and metabolomics, the interactive mechanisms of a sequential co-culture system of Trichoderma asperellum HG1 and Bacillus subtilis Tpb55 was explored in this study. The results revealed that co- culture has no significant effect on the growth and cell morphology of the two strains, but lead to mycelium wrinkling of HG1. RNA-seq analysis showed that co-culture significantly upregulated the HG1 genes concerning amino acid degradation and metabolism, proteolysis, resisting environmental stress, cell homeostasis, glycolysis, the glyoxylate cycle, and the citric acid (TCA) cycle, while Tpb55 genes related to cell homeostasis, spore formation and membrane fluidization were significantly upregulated, but genes associating to TCA, glycolytic cycles and fatty acid β -oxidation were significantly downregulated. Metabolomic results revealed that some amino acids related to energy metabolism were significantly altered in HG1, whereas palmitic acid, which is related to cell membrane functions, was upregulated in Tpb55. These results indicated that HG1 could interfere with carbon metabolism and cell membrane fluidity, but accelerate spore formation of Tpb55. Biophysical assays further convinced that co-culture could decrease ATP content and inhibit ATPase activity in HG1, and could promote spore formation and reduce the cell membrane fluidity of Tpb55. In addition, co-culture also accelerated the production of intracellular anti-oomycete compound octhilinone. The above results indicate that HG1 and Tpb55 are mainly in a competitive relationship in the co culture system. These findings provide new insights for understanding the interaction mechanism between co cultured microbes. [Display omitted] [ABSTRACT FROM AUTHOR]