Sulfur assimilation using gaseous carbonyl sulfide by the soil fungus Trichoderma harzianum.

Fungi have the capacity to assimilate a diverse range of both inorganic and organic sulfur compounds. It has been recognized that all sulfur sources taken up by fungi are in soluble forms. In this study, we present evidence that fungi can utilize gaseous carbonyl sulfide (COS) for the assimilation of a sulfur compound. We found that the filamentous fungus Trichoderma harzianum strain THIF08, which has constitutively high COS-degrading activity, was able to grow with COS as the sole sulfur source. Cultivation with 34S-labeled COS revealed that sulfur atom from COS was incorporated into intracellular metabolites such as glutathione and ergothioneine. COS degradation by strain THIF08, in which as much of the moisture derived from the agar medium as possible was removed, indicated that gaseous COS was taken up directly into the cell. Escherichia coli transformed with a COS hydrolase (COSase) gene, which is clade D of the ß-class carbonic anhydrase subfamily enzyme with high specificity for COS but low activity for CO2 hydration, showed that the COSase is involved in COS assimilation. Comparison of sulfur metabolites of strain THIF08 revealed a higher relative abundance of reduced sulfur compounds under the COS-supplemented condition than the sulfate-supplemented condition, suggesting that sulfur assimilation is more energetically efficient with COS than with sulfate because there is no redox change of sulfur. Phylogenetic analysis of the genes encoding COSase, which are distributed in a wide range of fungal taxa, suggests that the common ancestor of Ascomycota, Basidiomycota, and Mucoromycota acquired COSase at about 790-670 Ma.IMPORTANCEThe biological assimilation of gaseous CO2 and N2 involves essential processes known as carbon fixation and nitrogen fixation, respectively. In this study, we found that the fungus Trichoderma harzianum strain THIF08 can grow with gaseous carbonyl sulfide (COS), the most abundant and ubiquitous gaseous sulfur compound, as a sulfur source. When the fungus grew in these conditions, COS was assimilated into sulfur metabolites, and the key enzyme of this assimilation process is COS hydrolase (COSase), which specifically degrades COS. Moreover, the pathway was more energy efficient than the typical sulfate assimilation pathway. COSase genes are widely distributed in Ascomycota, Basidiomycota, and Mucoromycota and also occur in some Chytridiomycota, indicating that COS assimilation is widespread in fungi. Phylogenetic analysis of these genes revealed that the acquisition of COSase in filamentous fungi was estimated to have occurred at about 790-670 Ma, around the time that filamentous fungi transitioned to a terrestrial environment.

Thirteen new peptaibols with antimicrobial activities from Trichoderma sp.

From the fungus Trichoderma sp., we isolated seven novel 18-residue peptaibols, neoatroviridins E-K (1-7), and six new 14-residue peptaibols, harzianins NPDG J-O (8-13). Additionally, four previously characterized 18-residue peptaibols neoatroviridins A-D (14-17) were also identified. The structural configurations of the newly identified peptaibols (1-13) were determined by comprehensive nuclear magnetic resonance (NMR) and high-resolution electrospray ionization tandem mass spectrometry (HR-ESI-MS/MS) data. Their absolute configurations were further determined using Marfey's method. Notably, compounds 12 and 13 represent the first 14-residue peptaibols containing an acidic amino acid residue. In antimicrobial assessments, all 18-residue peptaibols (1-7, 14-17) exhibited moderate inhibitory activities against Staphylococcus aureus 209P, with minimum inhibitory concentration (MIC) values ranging from 8-32 µg·mL-1. Moreover, compound 9 exhibited moderate inhibitory effect on Candida albicans FIM709, with a MIC value of 16 µg·mL-1.

Exploration and Evaluation of Secondary Metabolites from Trichoderma harzianum: GC-MS Analysis, Phytochemical Profiling, Antifungal and Antioxidant Activity Assessment.

In this study, we investigated in vitro the potential of Trichoderma harzianum to produce bioactive secondary metabolites that can be used as alternatives to synthetic compounds. The study focused on analyzing two extracts of T. harzianum using ethyl acetate and n-butanol solvents with different polarities. The extracts were examined using phytochemical analysis to determine the content of polyphenols, flavonoids, tannins, and alkaloids. Thin-layer chromatography (TLC) and Gas chromatography-mass spectroscopy (GC-MS) analysis were used to profile volatile organic metabolites (VOCs) present in the extracts. Furthermore, the extracts were tested for their antifungal ability using the poison food technique. For measuring antioxidant activity, the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) test was used. Trichoderma harzianum was shown to have a significantly high content of tannins and alkaloids, with a noticeable difference between the two extracts. GC-MS analysis identified 33 potential compounds with numerous benefits that could be used in agriculture and the medicinal industry. Moreover, strong antifungal activity was identified against Sclerotinia sclerotiorum by 94.44%, Alternaria sp. by 77.04%, and Fusarium solani by 51.48; similarly, the IC50 of antioxidant activity was estimated for ethyl acetate extract by 71.47% and n-butanol extract by 56.01%. This leads to the conclusion that Trichoderma harzianum VOCs play a significant role as an antifungal and antioxidant agent when taking into account the advantageous bioactive chemicals noted in the extracts. However, to our knowledge, this is the first study in Algeria presenting detailed phytochemical analysis and GC-MS profiling of Trichoderma harzianum for two extracts, ethyl acetate and n-butanol.

Ameliorated Antibacterial and Antioxidant Properties by Trichoderma harzianum Mediated Green Synthesis of Silver Nanoparticles.

Biosynthesis of silver nanoparticles using beneficial Trichoderma harzianum is a simple, eco-friendly and cost-effective route. Secondary metabolites secreted by T. harzianum act as capping and reducing agents that can offer constancy and can contribute to biological activity. The present study aimed to synthesize silver nanoparticles using T. harzianum cell filtrate and investigate different bioactive metabolites based on LC-MS/MS analysis. The synthesized silver nanoparticles (AgNPs) from T. harzianum were characterized by ultraviolet-visible spectrophotometry, Fourier transform infrared spectrometry (FT-IR), energy-dispersive spectroscopy (EDS), dynamic light scattering (DLS), X-ray powder diffraction (XRD) and scanning electron microscopy (SEM). The surface plasmon resonance of synthesized particles formed a peak centered near 438 nm. The DLS study determined the average size of AgNPs to be 21.49 nm. The average size of AgNPs was measured to be 72 nm by SEM. The cubic crystal structure from XRD analysis confirmed the synthesized particles as silver nanoparticles. The AgNPs exhibited remarkable antioxidant properties, as determined by DPPH and ferric reducing antioxidant power (FRAP) assay. The AgNPs also exhibited broad-spectrum antibacterial activity against two Gram-positive bacteria (S. aureus and B. subtilis) and two Gram-negative bacteria (E. coli and R. solanacearum). The minimum inhibitory concentration (MIC) of AgNPs towards bacterial growth was evaluated. The antibacterial activity of AgNPs was further confirmed by fluorescence microscopy and SEM analysis.

The individual and interactive role of arbuscular mycorrhizal fungi and Trichoderma viride on growth, protein content, amino acids fractionation, and phosphatases enzyme activities of onion plants amended with fish waste.

The Green Revolution faced a great cost to meet ever-increasing demands for food, where indiscriminate use of agrochemicals resulted in non-friendly habitats. Therefore, the development of a sustainable approach to better crop production of onion seeds (Allium cepa L.) is very crucial. It is time to use organic waste as a replacement for agrochemicals by using arbuscular mycorrhizal fungi (AMF) and Trichoderma. Fish waste as representative of food waste acts as a leading cause of contamination of the environment. The interaction of AMF and Trichoderma viride on biomass, total soluble protein, mycorrhizal colonization, amino acids, phosphatases and phosphorus and nitrogen contents of onion plants grown in fish waste amended soil was studied. Fish waste has caused a slight increase in onions biomass, total free amino acids, and soluble protein content while with AMF and T. viride dual inoculation more increments were recorded; such increases were related to an increase in mycorrhizal colonization. T. viride application significantly increased the mycorrhizal colonization levels, but these were significantly reduced with waste addition. Analysis of amino acids in plants showed that their concentrations had changed as a result of waste addition combined with AMF and/or T. viride. The effectiveness of fish waste combined with low cost and health/environmental safety leads to a prediction that the introduction of fish waste coupled with fungi will become a more popular feature of agriculture in the future.

Trichoderma strains accelerate maturation and increase available phosphorus during vermicomposting enriched with rock phosphate.

AIMS: To suggest microbial inoculation as a tool to shorten organic residues stabilization and increase rock phosphate (RP) solubilization through vermicomposting, thus increasing nutrient content in plants and making it more appealing to farmers. Two Trichoderma strains were inoculated alone or combined in a RP apatite-enriched vermicompost. Stability and plant-available phosphorus levels were monitored for 120 days. METHODS AND RESULTS: Observable higher total organic carbon reduction in the treatment with the combined Trichoderma strains, followed by the inoculation with T. asperellum and T. virens. Combined Trichoderma and inoculation with T. virens increased humic acids (HA) content in 38·2 and 25·0%, respectively; non-inoculated vermicompost with T. asperellum increased it by 15·0%. The combined Trichoderma strains and T. virens achieved the stability index based on the humic/fulvic acids (HA/FA) ratio after 120 days. T. asperellum, combined Trichoderma and T. virens increased the citric acid soluble-P content in 83·2, 62·2 and 49·5%, respectively, compared to the non-inoculated vermicompost. CONCLUSIONS: Inoculation with combined T. asperellum and T. virens efficiently accelerated vermicompost stabilization; T. asperellum increased the citric acid soluble-P in the final product. SIGNIFICANCE AND IMPACT OF THE STUDY: Combined Trichoderma inoculation and RP enrichment improves the vermicompost quality, increasing HA and citric acid soluble-P, recycling organic waste nutrients and reducing agricultural dependence on phosphate fertilizers.

Trichoderma Volatile Organic Compounds as a Biofumigation Tool against Late Blight Pathogen Phytophthora infestans in Postharvest Potato Tubers.

We tested the ability of 14 strains of Trichoderma to emit volatile compounds that decreased or stopped the growth of Phytophthora infestans. Volatile organic compounds (VOCs) emitted from Trichoderma strains designated T41 and T45 inhibited the mycelial growth of P. infestans grown on a laboratory medium by 80 and 81.4%, respectively, and on potato tubers by 93.1 and 94.1%, respectively. Using the DNA sequence analysis of the translation elongation factor region, both Trichoderma strains were identified as Trichoderma atroviride. VOCs emitted by the strains were analyzed, and 39 compounds were identified. The most abundant compounds were 3-methyl-1-butanol, 6-pentyl-2-pyrone, 2-methyl-1-propanol, and acetoin. Electron microscopy of the hyphae treated with T. atroviride VOCs revealed serious morphological and ultrastructural damages, including cell deformation, collapse, and degradation of cytoplasmic organelles. To our knowledge, this is the first report describing the ability of Trichoderma VOCs to suppress the growth of the late blight potato pathogen.

Phosphorus-solubilizing Trichoderma spp. from Amazon soils improve soybean plant growth.

Acidic soils rapidly retain applied phosphorus fertilizers and consequently present low availability of this nutrient to plants. The use of phosphate-solubilizing microorganisms to help plant phosphorus (P) absorption is a promising sustainable strategy for managing P deficiencies in agricultural soils. Trichoderma strains have been one of the most studied filamentous fungi for improving the production and development of several crop species mainly due to their capability for symbiotic associations and their ability to control soil-borne plant diseases. Thus, this work sought to bioprospect Trichoderma strains from the Amazon rainforest capable of solubilizing/mineralizing soil phosphate and promoting soybean growth. Soybean plants inoculated with selected Trichoderma strains were cultivated in soil under greenhouse conditions and under a gradient of rock phosphate and triple superphosphate. As a result, 19.5% of the isolated Trichoderma strains were able to solubilize phosphate. In addition, those strains produced different organic acids during the solubilization process. Trichoderma spp. strains showed positive responses in the promotion of soybean growth-from 2.1% to 41.1%-as well as in the efficiency of P uptake-up to 141%. These results reveal the potential of Trichoderma spp. from the Amazon biome as promising biofertilizer agents.

Enhanced cellulase and reducing sugar production by a new mutant strain Trichoderma harzianum EUA20.

Trichoderma harzianum EU2-77 was a mutant strain of the wild-type strain T. harzianum NP13a isolated in Singapore. A multi-mutagenesis one-screening (MMOS) method was developed to further improve strain EU2-77 and a new mutant EUA20 was obtained. It exhibited filter paper cellulase (FPase) activity up to 14.79 IU/mL within 6 days shake flask cultivation. Activities of FPase, endoglucanase, ß-glucosidase, and xylanase, and protein content by EUA20 were respectively increased to 5.73, 4.35, 7.34, 1.80 and 2.70 folds. Using pretreated oil palm empty fruit bunch (OPEFB) and corncob powder as the substrates, strain EUA20 presented approximate 6.52 and 8.80 IU/ml FPase activity. Reducing sugar yield of 615.8 and 636.8 mg/g biomass were respectively obtained for OPEFB and corncob powder using cellulolytic enzymes of strain EUA20. Our results demonstrated that mutant strain EUA20 had great potential in on-site cellulase production for effective biomass bioconversion.

Enhanced anti-lung carcinoma and anti-biofilm activity of fungal molecules mediated biogenic zinc oxide nanoparticles conjugated with ß-D-glucan from barley.

This work reports the optimization, synthesis, characterization, anticancer, and antibacterial activity of the Trichoderma-ß-D-glucan­zinc oxide nanoparticles (T-ß-D-glu-ZnO NPs). Firstly, the T-ZnO NPs was synthesized using the fungal mycellial water extract (FWME) derived from T. harzianum (SKCGW009) under the optimized condition of extract concentration (5.99 mL), temperature (43.11 °C), pH (8) and time (69.04 h). The successful conjugation of T-ZnO NPs with ß-D-glucan (T-ß-D-glu-ZnO NPs) was confirmed by PACE and FTIR. The XRD, UHR SEM, and TEM EDS results pointed the spherical shape of NPs with the mean size of 30.34 nm. Further, the XPS survey scan and high-resolution fitting of Zn2p results also claimed the successful formation of the T-ß-D-glu-ZnO NPs. Cytotoxicity results indicated that the NPs were not toxic to NIH3T3 cells, while exhibited the dose-dependent inhibitory effect to human pulmonary carcinoma A549 cells. The IC50 of T-ZnO NPs and T-ß-D-glu-ZnO NPs against A549 cells was 158 and 56.25 µg.mL-1, respectively, which was also verified by fluorescent cytochemistry. Annexin V-FITC staining results indicated the presence of apoptotic cells in the NPs treated A549 cells, which was not seen in the non-treated control A549 cells. Interestingly, the number of necrosis cells was higher in the T-ZnO NPs (3.38%) comparing to T-ß-D-glu -ZnO NPs (0.07%). The early or late apoptosis was found higher in the cells treated T-ß-D-glu -ZnO NPs (6.43%) comparing with T-ZnO NPs (4%). These results indicated that T-ZnO NPs and T-ß-D-glu-ZnO NPs induced the cancer cell death through necrosis and apoptosis pathway, respectively. The antibacterial results indicated that the NPs treatment were significantly inhibited the growth of the Staphylococcus aureus inside of roundworm and enhanced growth of roundworm. Overall, anticancer and in vitro, in vivo antibacterial studies proved the high caliber of T-ß-D-glu-ZnO NPs for the further pharmaceutical evaluation.

Highly oxygenated polyketides produced by Trichoderma koningiopsis QA-3, an endophytic fungus obtained from the fresh roots of the medicinal plant Artemisia argyi.

Eight new highly oxygenated fungal polyketides, namely, 15-hydroxy-1,4,5,6-tetra-epi-koninginin G (1), 14-hydroxykoninginin E (2), koninginin U (3), 4'-hydroxykoninginin U (4), koninginin V (5), 14-ketokoninginin B (6), 14-hydroxykoninginin B (7), and 7-O-methylkoninginin B (8), together with six known related analogues (9-14), were isolated from Trichoderma koningiopsis QA-3, a fungus obtained from the inner root tissue of the well known medicinal plant Artemisia argyi. All these compounds are bicyclic polyketides, with compound 1 contains unusual hemiketal moiety at C-5 and compounds 2-14 having ketone group at C-1 and double bond at C-5(6). The structures and absolute configurations of the new compounds were established by spectroscopic analysis, X-ray crystal diffraction, modified Mosher's method, and ECD calculation. The absolute configurations of the known compounds 9, 10, and 12 were determined by X-ray crystal diffractions for the first time. The antimicrobial activities against human pathogen, marine-derived aquatic bacteria, and plant-pathogenic fungi of compounds 1-14 were evaluated, and compound 1 showed remarkable activity against aquatic pathogen Vibrio alginolyticus with MIC value 1 µg/mL, which is as active as that of the positive control.

Mycogenic Selenium Nanoparticles as Potential New Generation Broad Spectrum Antifungal Molecules.

The current challenges of sustainable agricultural development augmented by global climate change have led to the exploration of new technologies like nanotechnology, which has potential in providing novel and improved solutions. Nanotools in the form of nanofertilizers and nanopesticides possess smart delivery mechanisms and controlled release capacity for active ingredients, thus minimizing excess run-off to water bodies. This study aimed to establish the broad spectrum antifungal activity of mycogenic selenium nanoparticles (SeNPs) synthesized from Trichoderma atroviride, and characterize the bioactive nanoparticles using UV-Vis spectroscopy, dynamic light scattering (DLS), Fourier transform infrared (FT-IR), X-ray diffraction (XRD), scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS), and high-resolution transmission electron microscopy (HR-TEM). The synthesized nanoparticles displayed excellent in vitro antifungal activity against Pyricularia grisea and inhibited the infection of Colletotrichum capsici and Alternaria solani on chili and tomato leaves at concentrations of 50 and 100 ppm, respectively. The SEM-EDS analysis of the bioactive SeNPs revealed a spherical shape with sizes ranging from 60.48 nm to 123.16 nm. The nanoparticles also possessed the unique property of aggregating and binding to the zoospores of P. infestans at a concentration of 100 ppm, which was visualized using light microscope, atomic force microscopy, and electron microscopy. Thus, the present study highlights the practical application of SeNPs to manage plant diseases in an ecofriendly manner, due to their mycogenic synthesis and broad spectrum antifungal activity against different phytopathogens.

Enhanced tomato plant growth in soil under reduced P supply through microbial inoculants and microbiome shifts.

Soil microbial communities interact with roots, affecting plant growth and nutrient acquisition. In the present study, we aimed to decipher the effects of the inoculants Trichoderma harzianum T-22, Pseudomonas sp. DSMZ 13134, Bacillus amyloliquefaciens FZB42 or Pseudomonas sp. RU47 on the rhizosphere microbial community and their beneficial effects on tomato plants grown in moderately low phosphorous soil under greenhouse conditions. We analyzed the plant mass, inoculant colony forming units and rhizosphere communities on 15, 22, 29 and 43 days after sowing. Selective plating showed that the bacterial inoculants had a good rhizocompetence and accelerated shoot and root growth and nutrient accumulation. 16S rRNA gene fingerprints indicated changes in the rhizosphere bacterial community composition. Amplicon sequencing revealed that rhizosphere bacterial communities from plants treated with bacterial inoculants were more similar to each other and distinct from those of the control and the Trichoderma inoculated plants at harvest time, and numerous dynamic taxa were identified. In conclusion, likely both, inoculants and the rhizosphere microbiome shifts, stimulated early plant growth mainly by improved spatial acquisition of available nutrients via root growth promotion. At harvest, all tomato plants were P-deficient, suggesting a limited contribution of inoculants and the microbiome shifts to the solubilization of sparingly soluble soil P.

Upgrading the phenolic content, antioxidant and antimicrobial activities of garden cress seeds using solid-state fermentation by Trichoderma reesei.

AIMS: This study aims to evaluate the impact of solid-state fermentation (SSF) by Trichoderma reesei on the phenolic content, antioxidant and antimicrobial activities of garden cress seeds (GCS). METHODS AND RESULTS: The factorial statistical design was employed to optimize the SSF conditions, incubation time, pH, temperature and moisture, for maximum production of the phenolic content and microbial carbohydrate-cleaving enzymes from GCS. The total phenolic content significantly increased from unfermented GCS (401 mg gallic acid equivalent (GAE) 100 g-1 ) to fermented GCS (3600 mg GAE 100 g-1 ) by ninefold. The total antioxidant activity significantly increased in fermented GCS. Fifteen phenolic compounds were detected in fermented GCS with high concentrations compared to 14 in unfermented GCS using high-performance liquid chromatography. A strong correlation between the production of the carbohydrate-cleaving enzymes and the phenolic content of fermented GCS was observed. The phenolic compounds of fermented GCS showed higher antimicrobial activity. CONCLUSIONS: The fermented GCS is a powerful source of phenolic compounds with high antioxidant potentials, which can be used as dietary supplement and antimicrobial agent. SIGNIFICANCE AND IMPACT OF THE STUDY: Solid-state fermentation is a promising technique used for production of added-value bioactive compounds. SSF increased the total phenolic content and antioxidant activity of GCS several folds compared to germination process, which recently studied.

Bioaugmentation-assisted phytoremediation of lead and salinity co-contaminated soil by Suaeda salsa and Trichoderma asperellum.

The combined application of plant Suaeda salsa and indigenous fungus Trichoderma asperellum on the treatment of a lead (Pb) and salinity (Na+ and Ca2+) co-contaminated soil was investigated by a flowerpot experiment. As demonstrated by plant growth and selected antioxidant parameters, S. salsa was able to tolerate and grow in the co-contaminated soil, especially bioaugmented with T. asperellum, which promoted plant growth (9-23% and 5-13% increases for plant height and fresh weight, respectively) and appeared to alleviate plant oxidative damage (7-85% and 7-49% decreases for plant malondialdehyde and peroxidase levels, respectively). The SDS-PAGE fingerprints indicated that the total protein contents of S. salsa were affected under Pb and salinity stresses. The interactions of Na+ and Ca2+ ions on the phytotoxicity of Pb remained hormesis phenomenon that low-dose alleviation and high-dose enhancement. The analysis of phytoextraction parameters and bioavailability demonstrated that Pb was mainly concentrated in plant roots and poorly translocated, indicating the phytostabilization served as a major repair pathway. On the contrary, the Na+ and Ca2+ ions were concentrated in plant by the following order: shoot > root. Moreover, bioaugmentation of planted soil with T. asperellum generally led to the 9-42%, 13-58%, and 19-30% decreases of plant Pb, Na+, and Ca2+ concentrations and translocations, respectively, as well as a 6-21% decrease of soil Pb bioavailability. This study provided a bioaugmentation-assisted phytoremediation technique to make up the deficiencies of the long-term remediation for heavy metals and salinity.

Targeted metabolomic profiling reveals interspecific variation in the genus Hypericum in response to biotic elicitors.

Shoot cultures of eight Hypericum species belonging to the sections Hypericum, Oligostema, Ascyreia and Webbia were evaluated for their phytochemical profiles by high-performance liquid chromatography. In total, 17 secondary metabolites assigned to the groups of anthraquinones, phloroglucinols, hydroxycinnamic acids and flavonoids were detected. Furthermore, the elicitation potential of 18 biotic factors derived from saccharides, endophytic fungi and Agrobacterium rhizogenes was examined and statistically analysed with the paired two-sample t-test and principal component analysis. The production of naphthodianthrones and emodin was predominantly stimulated by elicitors derived from Fusarium oxysporum and Trichoderma crassum, while Piriformospora indica promoted the phloroglucinols production. Among flavonoids, the aglycone amentoflavone was readily increased by several elicitors up to 15.7-fold in H. humifusum treated by potato-dextrose broth. However, the chlorogenic acid proved to be the most susceptible metabolite to elicitation, when 31.7-times increase was detected in H. maculatum shoots upon D-glucose treatment. In spite of several biotic factors have been tested, no metabolite was commonly induced in all Hypericum spp. as a response to elicitor treatments.

An endophytic fungus from Trichoderma harzianum SWUKD3.1610 that produces nigranoic acid and its analogues.

The objective of this study was to determine whether endophytic fungi, isolated from Kadsura angustifolia produce nigranoic acid and its highly oxygenated derivatives. From the 426 endophytic fungi screened, Trichoderma harzianum SWUKD3.1610 was detected to have a component with the same TLC R f value and HPLC retention time as authentic nigranoic acid. This component was further confirmed as nigranoic acid by investigating the chemical composition of the fungal extracts. Besides (1), one new triterpenoid, 7ß- schinalactone C (2), and two known minor compounds were isolated and characterized by HRESIMS, 1D and 2D NMR spectroscopic methods. Our study indicates that endophytic fungus may play an important role in increasing the quality of the crude drugs from Chinese medicinal plant K. angustifolia. This study is the first to isolate, characterize, and identify schitriterpenes-producing Trichoderma spp.

Inhibitory effect of trichodermanone C, a sorbicillinoid produced by Trichoderma citrinoviride associated to the green alga Cladophora sp., on nitrite production in LPS-stimulated macrophages.

From the green alga Cladophora sp. collected in Italy, the marine fungal strain A12 of Trichoderma citrinoviride was isolated, identified and characterized. LC-MS qTOF analysis was applied to perform a metabolic profile of the fungal culture. Chromatographic techniques and spectroscopic methods were used to isolate and characterize the major secondary metabolites produced by this strain in liquid culture. In particular, four known sorbicillinoids (trichodermanone C, spirosorbicillinol A, vertinolide and sorbicillin) were purified and identified, together with 2-phenylethanol and tyrosol. Moreover, metabolomic analysis allowed to detect small amounts of trichodimerol, rezishanone A, 2',3'-dihydrosorbicillin and bisvertinol. For the first time a significant inhibitory effect on nitrite levels has been shown for trichodermanone C in lipopolysaccharide-stimulated J774A.1 macrophages.

The Role of Lignocellulosic Composition and Residual Lipids in Empty Fruit Bunches on the Production of Humic Acids in Submerged Fermentations.

The aim of this research was to study the production of humic acids (HA) by Trichoderma reesei from empty fruit bunches (EFBs) of palm oil processing, with a focus on the effects of lignocellulosic content and residual lipids. EFBs from two different soils and palm oil producers were previously characterized about their lignocellulosic composition. Submerged fermentations were inoculated with T. reesei spores and set up with or without residual lipids. The results showed that the soil and the processing for removal of the palm fresh fruits were crucial to EFB quality. Thus, EFBs were classified as type 1 (higher lignocellulosic and fatty acids composition similar to the palm oil and palm kernel oil) and type 2 (lower lignocellulosic content and fatty acids composition similar to palm oil). Despite the different profiles, the fungal growth was similar for both EFB types. HA production was associated with fungal growth, and it was higher without lipids for both EFBs. The highest HA productivity was obtained from type 1 EFB (approximately 90 mg L-1 at 48 h). Therefore, the lignocellulosic composition and the nature of the residual lipids in EFBs play an important role in HA production by submerged fermentation.

Duckweed (Lemna minor) is a novel natural inducer of cellulase production in Trichoderma reesei.

An inducer is crucial for cellulase production. In this study, duckweed was used as an inducer of cellulase production by Trichoderma reesei RUT C30. In a reaction induced by 50 g/L duckweed in shake flasks, the filter-paper activity (FPA) reached 6.5 FPU/mL, a value comparable to that induced by avicel. The enzyme-hydrolysis rate induced by steam-exploded corn stalk was 54.2%, representing a 28% improvement over that induced by avicel. The duckweed starch was hydrolyzed to glucose, which was subsequently used for biomass accumulation during the fermentation process. Furthermore, to optimize the control of the fermentation process, a combined substrate of avicel and duckweed was used to induce cellulase production by T. reesei RUT C30. The cellulase production and hydrolysis rates of the combined substrate, compared with avicel alone, were 39.6% and 36.7% higher, respectively. The results of this study suggest that duckweed is a good inducer of cellulase production in T. reesei, and it might aid in decreasing the cost of lignocellulosic materials hydrolysis.