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The present study aimed to purify and identify the metabolites from T. atroviride using high-performance liquid chromatography (HPLC) and 1 H and 13 C nuclear magnetic resonance spectrometer (NMR) followed by analyzing their toxicological, antibacterial and anticancer properties. This work identified two metabolites - TM1 and TM2. TM1 was in two forms: (i) 1, 3-dione-5, 5-dimethylcyclohexane; and, (ii) 2-enone-3hydroxy -5,5-dimethylcylohex, while TM2 was 4H-1,3-dioxin-4-one-2,3,6-trimethyl. These metabolites did not exhibit any irritant or allergic reaction as revealed by HET- CAM test. TM2 significantly inhibited the growth of H. pylori and Shigella toxin producing Escherichia coli (STEC) as evident by in vitro and microscopic observations of bacterial cell death. TM2 also induced the cell death and cytotoxicity, as revealed by cell viability test and western blot analysis. According to microscopic, flow cytometer and western blot analysis, TM2 treated cells displayed higher ROS, cell death, and apoptosis-related protein expression than TM1 and control. This study concluded that TM2 derived from T. atroviride was a potential therapeutic agent for anti-prostate cancer and antibiotic agent against MDR- H. pylori and STEC and it is also recommended to carry out further in vivo animal model experiments with improved stability of the metabolites for future pharmaceutical trails., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This paper reports the synthesis of chitosan nanoparticles (T-CSNPs) using the fungal enzyme of Trichoderma harzianum and its biocompatibility, antioxidant and bactericidal properties. The T-CSNPs synthesis was confirmed by absorbance at 280 nm using UV-Vis spectrophotometer. T-CSNPs were of spherical shape, as evident by field emission transmission electron microscopic (FETEM) analysis, and the average size of T-CSNPs was 90.8 nm, as calculated using particle size analyzer (PSA). The functional groups showed modifications of chitosan in T-CSNPs as evident by fourier-transform infrared spectroscopic (FTIR) analysis. T-CSNPs were found soluble at the wide range of pH, showing 100% solubility at pH 1-3 and 72% at pH 10. The T-CSNPs exhibited antioxidant property in a dose-dependent manner with pronounced activity at 100 mg·mL -1 . The T-CSNPs also showed bactericidal activity against Staphylococcus aureus and Salmonella enterica Typhimurium by causing detrimental effects on bacterial cells. The T-CSNPs (50 μg·mL -1 ) did not display any cytotoxic effect on murine fibroblast NIH-3T3 cells, as evident by cell viability and acridine orange/ethidium bromide staining assays, which confirmed biocompatibility of the nanoparticles. This work suggested further investigations on the utilization of the mycosynthesized nanochitosan in biomedical applications., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with 95.4 ± 0.61% of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at 500 μg ml -1 showed the total antioxidant of 48.70 ± 2.90%, DPPH radical scavenging activity of 37.25 ± 2.25, nitric oxide (NO) radical scavenging activity of 54.55 ± 1.95 and H 2 O 2 radical scavenging activity of 43.75 ± 3.21. The F41 at 25 μg ml -1 displayed antibacterial activity against E. coli (14.25 ± 0.25 mm), Proteus mirabilis (10.40 ± 0.60 mm), and Enterobacter aerogenes (5.60 ± 0.40 mm). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) in the F41, and hence these fatty acids are likely responsible for the antioxidant and antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells., (Copyright © 2018. Published by Elsevier Ltd.)

Antibacterial silver nanoparticles (AgNPs) are synthesized from various biological resources, but studies on Trichoderma-based synthesis are limited. Hence, the current work synthesized antibacterial silver nanoparticles using Trichoderma atroviride and also tested for their biomedical properties. AgNPs synthesis was confirmed by the occurrence of plasmon resonance at 390-400 nm in the UV-vis spectrum. FTIR spectrum displayed the bands at 1115.4 and 3450 cm -1 indicating the vibrations of the metallic oxides. Transmission electron microscopic and EDX analysis confirmed a high percentage signal of anisotropic structural AgNPs with the size of 15-25 nm. The AgNPs exhibited high antibacterial activity and DPPH scavenging activity in a dosedependent manner. The cytotoxicity assay \\\\ indicated that the AgNPs triggered the cancer cell death at the inhibitory concentration of IC 50 16.5 μg/ml. Current work reported for the first time Trichoderma atroviride based synthesis of anisotropic structural AgNPs with the relatively small size of 15-25 nm with remarkable anti-bacterial, antioxidant and cytotoxicity activities, and this deserves future investigations for biomedical applications., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Fusarium stalk rot (FSR) caused by Fusarium graminearum (FG) significantly affects the productivity of maize grain crops. Application of agrochemicals to control the disease is harmful to environment. In this regard, use of biocontrol agent (BCA) is an alternative to agrochemicals. Although Trichoderma species are known as BCA, the selection of host-pathogen specific Trichoderma is essential for the successful field application. Hence, we screened a total of 100 Trichoderma isolates against FG, selected Trichoderma harzianum (CCTCC-RW0024) for greenhouse experiments and studied its effect on changes of maize rhizosphere microbiome and biocontrol of FSR. The strain CCTCC-RW0024 displayed high antagonistic activity (96.30%), disease reduction (86.66%), biocontrol-related enzyme and gene expression. The root colonization of the strain was confirmed by eGFP tagging and qRT-PCR analysis. Pyrosequencing revealed that exogenous inoculation of the strain in maize rhizosphere increased the plant growth promoting acidobacteria (18.4%), decreased 66% of FG, and also increased the plant growth. In addition, metabolites of this strain could interact with pathogenicity related transcriptional cofactor FgSWi6, thereby contributing to its inhibition. It is concluded that T. harzianum strain CCTCC-RW0024 is a potential BCA against FSR.

To investigate the biodiversity of Trichoderma (Hypocreaceae) and their relation to sediment physical and chemical properties, we collected a total of 491 sediment samples from coastal wetlands (tidal flat and wetland) in Southeast China. Further, we applied two types of molecular approaches such as culture dependent and independent methods for identification of Trichoderma spp. A total of 254 isolates were obtained and identified to 13 species such as T. aureoviride, T. asperellum, T. harzianum, T. atroviride, T. koningiopsis, T. longibrachiatum, T. koningii. T. tawa, T. viridescens, T. virens, T. hamatum, T. viride, and T. velutinum by the culture-dependent (CD) method of these, T. tawa was newly described in China. Subsequently, the culture indepented method of 454 pyrosequencing analysis revealed a total of six species such as T. citrinoviride, T. virens, T. polysporum, T. harzianum/Hypocrea lixii and two unknown species. Notably, T. citrinoviride and T. polysporum were not found by the CD method. Therefore, this work revealed that the combination of these two methods could show the higher biodiversity of Trichoderma spp., than either of this method alone. Among the sampling sites, Hangzhou, Zhejiang Province, exhibited rich biodiversity and low in Fengxian. Correlation and Redundancy discriminant analysis (RDA) revealed that sediment properties of temperature, redox potential (Eh) and pH significantly influenced the biodiversity of Trichoderma spp., Competing Interests: The authors have declared that no competing interests exist.

Trichoderma harzianum is well known to exhibit induced systemic resistance (ISR) to Curvularia leaf spot. We previously reported that a C6 zinc finger protein (Thc6) is responsible for a major contribution to the ISR to the leaf disease, but the types of effectors and the signals mediated by Thc6 from Trichoderma are unclear. In this work, we demonstrated that two hydrolases, Thph1 and Thph2, from T. harzianum were regulated by Thc6. Furthermore, an electrophoretic mobility shift assay (EMSA) study revealed that Thc6 regulated mRNA expression by binding to GGCTAA and GGCTAAA in the promoters of the Thph1 and Thph2 genes, respectively. Moreover, the Thph1 and Thph2 proteins triggered the transient production of reactive oxygen species (ROS) and elevated the free cytosolic calcium levels in maize leaf. Furthermore, the genes related to the jasmonate/ethylene signaling pathway were up-regulated in the wild-type maize strain. However, the ΔThph1- or ΔThph2-deletion mutants could not activate the immune defense-related genes in maize to protect against leaf disease. Therefore, we conclude that functional Thph1 and Thph2 may be required in T. harzianum to activate ISR in maize.

• Cytotoxicity of aptamer (APT) tagged fungal crude protein capped AuNPs was analyzed. • The successful formation of the APT tagged AuNPs was evidenced by AGE. • APT- FE- AuNPs (521) exhibited higher cytotoxicity in A549 cells than other AuNPs. • APT- FE-AuNPs (521) triggered cytotoxicity through induction of oxidative stress. The present work demonstrates how the fungal crude protein extract (FE) mediated gold nanoparticles (AuNPs) functionalized with aptamer (APT) improved cytotoxicity in human lung cancer cell line (A549) and brain tumor cell line (LN229). The agarose gel electrophoresis (AGE) confirmed the conjugation of APT in FE-AuNPs. The FTIR results revealed the involvement of FE in the amalgamation of FE-AuNPs. FE-AuNPs (λ max 521 nm) had Z-average size of 19.72 ± 0.26 d. nm and PDI of 0.58 ± 0.01 with a zeta potential of -22.20 mV, whereas APT-FE-AuNPs (λ max 521 nm) had the Z-average size of 167.63 ± 1.40 d. nm and PDI of 0.21 ± 0.01 with a zeta potential of -36.36 mV. Both synthesized nanoparticles exhibited less cytotoxicity in normal NIH3T3 cells, while the APT-FE-AuNPs (λ max 521 nm) showed significant cytotoxicity in A549 and LN229 cells. The cells treated with APT-FE-AuNPs (λ max 521 nm) displayed higher percentage of early apoptosis (6.97 %), apoptosis (7.08 %), and necrosis (6.63 %) than those treated with bare or APT-FE-AuNPs (λ max 565 nm) in A549 cells. The results demonstrated that the DNA aptamer (AS1411) tagged with FE-AuNPs (λ max 521 nm) increased their accumulation in cancer cells, which triggered nucleolin targeted cell cytotoxicity through nucleus damage and induction of oxidative stress. [ABSTRACT FROM AUTHOR]

• pH-dependent polymer based drug delivery system (DDS) for fungal metabolites (TM2). • TM2 loaded CNPs was prepared, and pH-dependent release was characterized. • Chitosan (CS) based DDS improved the anticancer activity of TM2. • TM2-CNPs caused PC-3 cell death by regulating the apoptosis-related proteins. Trichoderma species are known to produce the molecules with promising anticancer, antimicrobial, and antiviral activities. Likewise, we previously reported a metabolite (TM2) from T. atroviride as an anti-prostate cancer agent but its activity was limited due to less ability to target the cancer cells. Therefore, the present work aimed to load the different concentrations of TM2 (1.25−20 mg.mL−1) in chitosan nanoparticles (TM2-CNPs) to improve its cytotoxicity in PC3 cells. The input of 10 mg.mL−1 of TM2 was achived spherical shaped TM2-CNPs with good drug entrapment efficiency of 50.97 ± 0.12% and drug loading capacity of 24.68 ± 0.64%. The TM2-CNPs exhibited an average size of 187.8 ± 1.51 nm, PDI of 0.197 ± 0.02, and zeta potential of 21.4 ± 0.17 mV. The FTIR spectra showed the functional group (NH 2 and O H) of chitosan in TM2-CNPs. The pH-dependent release was found >50% in the pH 5.8 while < 10% in pH 7.4. The IC50 dosage of TM2-CNPs treatment was caused 60.48% cell death in the PC3 cells through activation of caspase 3 and inhibition of the BCL-2 expression. Overall, this work has demonstrated that the pH-sensitive release of TM2 from nanocarrier (CNPs) increased anti-prostate cancer activity. [ABSTRACT FROM AUTHOR]

Abstract In this report, copper oxide nanoparticles (TA-CuO NPs) were synthesized using cell-free extract of Trichoderma asperellum and assessed their photothermal induced anticancerous activity. The fungal mediated TA-CuO NPs was confirmed by the surface plasmon resonance at 285–295 nm. The amide (C O) and aromatic (C C) groups in secondary metabolites of the extract was found to be an encapsulating or reducing agents for TA-CuO NPs, as indicated by IR spectra. Crystalline nature by cubic face-centered structure of the TA-CuO NPs was confirmed by XRD and their size ranges from 10 to 190 nm and an average of 110 nm by particle size analyzer (PSA). The Ultra HRSEM study revealed spherical shaped TA-CuO NPs. The FETEM results were also in strong agreement with PSA and UHR SEM. The survey-scan spectrum of XPS indicated the presence of C1s (47.83%), Cu2p (16.11%), Na1s (2.2%) and O1s (33.86%). The cell death was significantly found higher in photothermal induced by near-infrared laser (TA-CuO NPs-NIR) treated than that of TA-CuO NPs treatment. The level of ROS (35.62%) was higher in the treated cells than that of the untreated control, in accordance with the nucleus damage and losses in the mitochondrial membrane potential (ΔΨm). The upregulation of Bcl-2 in the untreated cells and Cas-3 in TA-CuO NPs-NIR treated cells was confirmed by western blot analysis. This work agreed with the potential biogenic TA-CuO NPs for promising in vitro photothermolysis of cancer cells, for the development of anticancer nanotherapeutics. Graphical Abstract Unlabelled Image Highlights • CuO NPs were synthesized from aqueous extracts of Trichoderma sp. • High through-put techniques were adopted for characterization of CuO NPs. • The synthesized CuO NPs found to have photothermolysis of A549 cancer cells. [ABSTRACT FROM AUTHOR]

Abstract We earlier reported the role of cellulase (Thph1 and Thph2) from Trichoderma harzianum in controlling the Curvularia leaf spot and Fusarium stalk rot through interaction with the maize root. But, the maize root receptor related to cellulase was unknown. Hence, in the present study, we applied label-free proteomics and yeast two-hybrid (Y2H) assay to screen the root receptor that communicates with cellulase by generating a different combination of activation (AD) and binding (bait) domain plasmids such as DDO-SD/-Leu/-Trp QDO/X-SD/-Ade/-His/-Leu/-Trp. The maize roots were treated with T. harzianum strains including knockout Thph1, Thph2 and wild-type and enzyme cellobiohydrolase 1 (CBH1). The treated or untreated maize roots were collected for extraction of RNA and protein for generation of cDNA library and Label-free proteomics assay respectively. The label-free proteomic results indicated that the protein expression in the maize root significantly varied between the treatments (P < 0.05) and the protein clusters were highly expressed in the treatments of T2 (inoculated with one g of conidia of T. harzianum WT Th22), T3 (inoculated with one g of conidia of knockout strains of Thph1 and Thph2), T4 (inoculated with enzyme CBH 1) as compared to T1 (uninoculated control). The Y2H screening results indicated that autophagocytosis associated protein (ZmATG3) and germin-like protein (ZmGLP) from the maize root seemed to communicate with cellulase Thph1 and Thph2 respectively. The present screening results deserve for further cellular localization of ZmATG3 and ZmGLP in the maize root and their single or synergic functioning in induced systemic resistance (ISR) in maize through molecular dialogue between Trichoderma spp. and maize roots. Graphical abstract Image 1 Highlights • Cellulase (Thph1 and Thph2) from Trichoderma regulates the maize roots proteins. • The interactions of Trichoderma elicitor and maize root receptor were screened by Y2H assay. • Autophagocytosis protein from the maize roots interact with cellulase Thph1. • Germin-like protein from the maize roots interact with cellulase Thph 2. [ABSTRACT FROM AUTHOR]

Abstract This work reports the role of cellulase from Trichoderma harzianum strain Th22 in stimulation of 2,4-Dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) and defense-related gene expressions in maize root against deoxynivalenol producing Fusarium graminearum (FG) by in vitro, in planta and virtual molecular modeling experiments. The results indicated that cellulase genes such as Thph1 & 2 were involved in biocontrol of FG and its related plant diseases through the activation of JA, ET, SAR, DIMBOA and PII related gene expressions in the maize root as evidenced by qRT-PCR and HPLC analysis. Further, the inhibitory assay also confirms that DIMBOA inhibited the growth of FG at concentrations dependent manner. Furthermore, virtual studies revealed that DIMBOA from maize root metabolizes or inhibit the active site of pathogenicity (FgSwi6) and mycotoxin (FgAtf1) related protein in FG. Finally, this work concluded that cellulase genes Thph1 and Thph2 from T. harzianum was not only limited to control of foliar disease but also involved in biocontrol of Fusarium stalk rot. Graphical abstract Image 1 Highlights • Cellulase genes triggering plant defense against pathogens F. graminearum (FG). • Cellulase involved in synthesis and expression of DIMBOA. • Cellulase activated DIMBOA inhibited the FG. • DIMBOA inhibits pathogencity and mycotoxin related proteins in pathogen FG. • Cellulase from T. harzianum proved as involved in control of Fusarium stalk rot. [ABSTRACT FROM AUTHOR]

The efficacy of seven strains of Trichoderma asperellum collected from the fields in Southern China was assessed against Fusarium graminearum (FG) the causal agent of corn stalk rot of maize were in vitro for their antagonistic properties followed by statistical model of principal compound analysis to identify the beneficial antagonist T. asperellum strain. The key factors of antagonist activity were attributed to a total of 13 factors including cell wall degrading enzymes (chitnase, protease and β-glucanases), secondary metabolites and peptaibols and these were analyzed from eight strains of Trichoderma. A linear regression model demonstrated that interaction of enzymes and secondary metabolites of T. asperellum strain ZJSX5003 enhanced the antagonist activity against FG. Further, this strain displayed a disease reduction of 71 % in maize plants inoculated with FG compared to negative control. Pointing out that the T. asperellum strain ZJSX5003 is a potential source for the development of a biocontrol agent against corn stalk rot. [ABSTRACT FROM AUTHOR]

Cucumber ( Cucumis sativus L.) is an important vegetable crop grown worldwide. However, its productivity is largely suppressed the phytopathogen Fusarium oxysporum f. sp. cucumerinum (FOC). We screened 100 isolates of Trichoderma as potential biocontrol agents against FOC. Both in vitro and in vivo methods were used, followed by the analysis of synergistic effect of cell wall degrading enzymes (CWDEs) and its secondary metabolites. In addition, molecular target of growth regulating protein of FOC (Snt2, a BAH/PHD containing transcription factor) was also studied. Of the 100 isolates of Trichoderma , 10 isolates with more than 85% inhibition rate were selected for in vivo antagonism in greenhouse experiment. Trichoderma asperellum strain CCTCC-RW0014 was found to have good biocontrol potential with high disease reduction of 71.67%. The RT-qPCR analysis revealed that the ITS copies of T. asperellum CCTCC-RW0014 were significantly higher in the cucumber roots treated with T. asperellum CCTCC-RW0014. Whereas the ITS copies of FOC declined in plants root in the presence of T . asperellum CCTCC-RW0014 and FOC. T . asperellum CCTCC-RW0014 showed high hydrolytic activity of chitin (87.5 ± 3.21%), gelatin (84.8 ± 2.56%), carboxymethyl cellulose (52.9 ± 1.23%) and pachyman (60.5 ± 2.32%). The crude enzymes; namely chitinase, cellulase, protease, and β (1–3) glucanase; were optimally active at 40 °C and pH 5.0, 30 °C and pH 6.0, 40 °C and pH 7, and 40 °C and pH 6.0 respectively. The secondary metabolites of Trichoderma extracts were determined by GC–MS and further studied for the molecular docking against target protein Snt2. Among the tested compounds, 1, 6-diphenylhexane-1,3,4,6-tetrone showed the highest C dock energy (−27.567 kacl/mol) against the target protein and this may down regulate the expression of protein Snt2. The present work concluded that T . asperellum CCTCC-RW0014 was the highly potential strain to control the FOC, providing scope for a further study on the mechanism of interaction between Trichoderma derived compounds and host plants. [ABSTRACT FROM AUTHOR]

The present work aimed to use the biogenic metallic nanoparticles as catalyst for bioelectricity production in microbial fuel cell (MFC) approach under anaerobic condition. Silver and gold nanoparticles (AuNPs) were synthesized using Trichoderma sp. Particle size and cystallinity were measured by X-ray diffraction revealed the crystalline structure with average size of 36.17 nm. Electron microscopic studies showed spherical shaped silver nanoparticles (AgNPs) and cubical shaped AuNPs with size ranges from 50 to 150 nm. The concentration of biogenic metallic nanoparticles as catalyst for enhanced bioelectricity generations and estimated by response surface methodology (RSM) and found at the greatest of 342.80 mA under optimized conditions are time interval, temperature, nanoparticles used as 63 h, 28 ± 2.0 °C, 22.54 mg l −1 (AgNPs) and 25.62 mg l −1 (AuNPs) in a batch reactor. AuNPs acted as an excellent catalyst to enhance the bioelectricity production. This novel technique could be used for eco-friendly, economically feasible and facile electricity production. [ABSTRACT FROM AUTHOR]

The present work aimed at purifying the intracellular fungal metabolites, such as 16-methylheptadecanoic acid methyl ester (HDA) and 9,12-octadecadienoic acid (ODA) from marine Trichoderma , Hypocrea lixii TSK8, Hypocrea rufa SKS2 respectively, and investigating their anticancer and antioxidant effects. The two fungal metabolites were tested against two human cancer cell lines, namely oral cancer (KB) and skin carcinoma (A431) by using MTT assay. The inhibitory concentrations (IC 50 ) against KB oral cancer cells were found to be 18.75 ± 0.12 μg/mL for HDA and 75.50 ± 0.42 μg/mL for ODA. Whereas IC 50 values of HDA and ODA against A431 were found 37.5 ± 0.42 μg/mL and 72.89 ± 0.15 μg/mL, respectively. In addition, the down-regulation of heat shock protein 90 kDa (HSP90) was confirmed by using SDS-PAGE and Western blot analysis. The effect of HDA induced apoptosis via ROS-dependent internucleosomal DNA fragmentation was confirmed by AGE analysis. We further evaluated the in vivo anti-skin cancer activity of HDA in Swiss albino mice induced with skin cancer by 7,12-dimethylbenz(a)anthracene (DMBA) and croton oil (CO). The in vivo hematological, biochemical and histopathological results revealed that the fungal metabolite HDA was a highly potent anticancer compound against the skin cancer. [ABSTRACT FROM AUTHOR]

In this work, a total of six polysaccharides were isolated from culture filtrate (EPS1, EPS2) and mycelia (IPS1–IPS4) of Trichoderma harzianum. The HPLC analysis results showed that EPS1, EPS2, IPS1, and IPS2 were composed of mannose, ribose, glucose, galactose, and arabinose. The FT-IR, 1H, and 13C NMR chemical shifts confirmed that the signals in EPS1 mainly consist of (1→4)-linked α-d-glucopyranose. EPS1 and IPS1 showed a smooth and clean surface, while EPS2, IPS2, and IPS3 exhibited a microporous structure. Among polysaccharides, EPS1 displayed higher ABTS+ (47.09 ± 2.25% and DPPH (26.44 ± 0.12%) scavenging activities, as well as higher α-amylase (69.30 ± 1.28%) and α-glucosidase (68.22 ± 0.64%) inhibition activity than the other polysaccharides. EPS1 exhibited high cytotoxicity to MDA-MB293 cells, with an IC50 of 0.437 mg/mL, and this was also confirmed by cell staining and FACS assays. These results report the physicochemical and bioactive properties of polysaccharides from T. harzianum. [ABSTRACT FROM AUTHOR]

This work reports the optimization, synthesis, characterization, anticancer, and antibacterial activity of the Trichoderma -β-D-glucan‑zinc oxide nanoparticles (T-β-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 β-D-glucan (T-β-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-β-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 IC 50 of T-ZnO NPs and T-β-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-β-D-glu -ZnO NPs (0.07%). The early or late apoptosis was found higher in the cells treated T-β-D-glu -ZnO NPs (6.43%) comparing with T-ZnO NPs (4%). These results indicated that T-ZnO NPs and T-β-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-β-D-glu-ZnO NPs for the further pharmaceutical evaluation. Unlabelled Image • We report the synthesis and bioactivities of T-β-D-glu-ZnO NPs. • High standard characterization methods applied to investigate the T-β-D-glu-ZnO NPs. • Anti-lung cancer activity of these NPs in A549 cells was evidenced by flow cytometer. • T-β-D-glu-ZnO NPs exhibited excellent antibacterial activity. [ABSTRACT FROM AUTHOR]

Trichoderma species are known to protect the plants from pathogen infections through multifunctions, such as secondary metabolism, mycoparasitism, hyperparasitism, nutrient competition, enzymes and induced systemic resistance (ISR). Herein, we isolated a total of 18 Trichoderma strains divided to nine species such as T. atroviride, T. virens, T. velutinum, T. harzianum, T. asperellum, T. koningiopsis, T. aureoviride, H. lixii, and T. koningii from the soils samples, collected from the wetland ecosystem of South Korea. These strains were screened against the pathogens- Macrophomina phaseolina (MP) , Fusarium graminearum (FG), and Botrytis cinerea (BC) - by in vitro antagonistic assay. Amongst, T. aureoviride (SKCGW013) showed higher antagonistic activity against the targeted pathogens than other isolates did. The strain SKCGW013 was further used for extraction, purification and analysis of the metabolites by column chromatography (CC) and gas chromatography mass spectroscopy (GC-MS). The expression of secondary metabolites regulatory genes of non-ribosomal peptide synthetase (NRPS), polyketide synthase (PKS) were studied by RT-qPCR. The results showed the presence of eight dominant compounds in the ethyl acetate fraction of the strain SKCGW013 and these compound were then screened by molecular modeling method against phytopathogens. In addition, RT-qPCR study revealed the significant expression of metabolites related genes. Further molecular docking study showed that the compounds from strain SKCGW13 synergistically inhibited the targeted pathogens. Among the compounds - 2H-Pyran, 3-bromo-2-butoxytetrahydro-, cis - exhibited high docking inhibitory energy against the targeted proteins, FgSwi6 and Bcpmr1 from FG and BC respectively. Overall, this study concluded that T. aureoviride SKCGW013 was an excellent source for discovery of novel metabolites as bio-control agents as evident by its metabolite profile with antifungal activity. Image 1 • This work reported the potent biocontrol strain from wetland soil of Republic of Korea. • A total of nine Trichoderma species was isolated with potent biocontrol properties. • T. auroviride (SKCGW013) inhibited the growth of various phytopathogens. • Metabolites from the T. auroviride induced biocontrol activity synergistically. [ABSTRACT FROM AUTHOR]