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Nitrogen is essential for plants’ growth, yield, and crop quality, and its deficiency limits food production worldwide. In addition, excessive fertilization and inefficient use of N can increase production costs and cause environmental problems. A possible solution to this problem is the application of biofertilizers, which improve N assimilation and increase biomass and yield. Therefore, the objective of this research was to evaluate the impact of the application of a combination of green and red algae (Ulva lactuca and Solieria spp.), Rhizobium sp., Trichoderma asperellum, and the combination of the above three biofertilizers on N assimilation. A completely randomized design was performed, with 10 plants per treatment and five treatments: T1 = control; T2 = algal extracts; T3 = Rhizobium sp.; T4 = T. asperellum; T5 = T2 + T3 + T4. Our analyses showed that the biofertilizers’ application was better than the control. The application of Rhizobium sp. had the best performance amongst all of the biofertilizers, with the highest nitrate reductase activity in maize leaves, which enhanced photosynthesis, increasing biomass and yield. The use of Rhizobium sp. showed increases in biomass (13.4%) and yield (11.82%) compared to the control. This research shows that biofertilizers can be a key component for sustainable agricultural practices.

Field trials were conducted for two seasons in two experimental sites (Mwea in Kirinyaga and Ngoliba in Kiambu counties of Kenya) to assess the efficacy of fungal endophytes Hypocrea lixii F3ST1 and Trichoderma asperellum M2RT4 in the control of Trialeurodes vaporariorum vector of tomato infectious chlorosis virus (TICV) and tomato chlorosis virus (ToCV) through seeds inoculation. TICV and ToCV’s disease incidence, severity and the yield were also evaluated. All the fungal endophytes successfully colonized all the tomato plant parts, but the highest root colonization was observed in H. lixii F3ST1 compared to the T. asperellum M2RT4 in both seasons. The number of nymphs was significantly lower in the endophytically colonized tomato plants than the control treatments in all the seasons and at both sites. However, the lowest number of nymphs was recorded in H. lixii F3ST1 compared to T. asperellum M2RT4. On the other hand, the TICV and ToCV disease incidence and severity rates were lower in endophytically colonized tomato crops compared to the control plots. This could be attributed to the reduction in the virus replication and lower feeding ability of T. vaporariorum that was characterized by less excretion of honeydew causing sooty mold. However, no significant difference was observed in ToCV disease severity rates among the treatments and across the seasons. The yield was significantly higher in endophyte plots than the control treatments in both sites and across the two seasons. This study demonstrates that H. lixii F3ST1 and T. asperellum M2RT4 endophytically colonized tomato plants and conferred systemic resistance against T. vaporariorum vector, and significantly reduced the transmission of TICV and ToCV, contributing to high reduction of both diseases’ incidence and severity in the field. However, further studies are warranted to confirm these results at large scale trials.

Abstract The experiment was conducted at the Department of Plant Pathology, Bangladesh Agricultural University, Mymensingh to identify T. asperellum in a countrywide screening program and to evaluate its antagonistic effect against several soil borne pathogens. Samples were collected from the rhizosphere soil of 49 different crops in 107 different locations in Bangladesh, especially, considering the several isolates of T. asperellum for purification. Based on morphological and physiological features, fifteen isolates were selected. Of these, the isolates of TR27 and TR45 were grown and sporulated at 40 °C except all the isolates with 35 °C, and particularly, showing a decrease of mycelial growth across all the isolates for increasing pH. Meanwhile, T. asperellum showed significant antagonistic effects against Fusarium oxysporum, Sclerotium rolfsii and Pythium aphanidermatum, resulting in reducing foot and root rot, collar rot and damping off diseases, respectively. Four isolates were selected for molecular characterization among 15 isolates in terms of higher mycelial growth and spore density in-vitro condition, isolates of (TR27) Sadar, Moulvibazar (Rice), (TR45) Sadar, Mymensingh (Sweet gourd), (TR70) Chapra, Chapai Nawabganj (Sesame) and (TR85) Nayanpur, Lalmonirhat (Maize) were studied at ITS and TEF region. Isolates of TR45, TR70 and TR85 were observed with 98% homology, and TR27 exhibited 88% in their respective closest isolates at ITS sequences. Isolates of TR27 and TR85 also exerted their respective nearest homology (96%), while TR45 showed 99%, and 93% homology with TR70 in TEF sequences. Isolates TR45, TR70 and TR85 were evidently determined as T. asperellum of 100% bootstrap value, and TR27 isolate was also recognized with 72% bootstrap value in the phylogenetic tree. However, complementary effects of significant superior homology and the greatest bootstrap value in the identification of T. asperellum were found as noteworthy. In the phylogenetic analysis, magnificent differentiation among the Trichoderma isolates within and among the groups of closely related species was observed in Tef1 region than reflecting maximum variability in the isolates of rDNA at ITS region, whereas demonstrating a higher transversion ratio and evolutionary divergence.

An effective method for maintaining the activity and longevity of microorganisms in adverse conditions is microencapsulation. In the present study, synthetic alginate pellets were developed as carriers for the biocontrol agent Trichoderma asperellum. In two field experiments, solarization was applied for three weeks to loamy clay soil that was naturally infested with strawberry-black root rot fungi (Fusarium solani, Rhizoctonia solani and Machrophomina phaseolina). Following solarization, T. asperellum-based alginate pellets or spore suspension were added to the soil. Data reveal that, three weeks solarization of irrigated soil increased its maximum temperature reached by 11–14.2 °C (1–10 cm depth), 11.6–13.1 °C (11–20 cm depth) and 10.1–12.2 °C (21–30 cm depth). In either trial, solarization also successfully lowers the vitality of strawberry-black root rot fungi directly after the solarization phase. When compared to controls, strawberry-black root rot was substantially less common in solarized plots. In two field trials, soil solarization followed by inoculation with alginate pellets based on T. asperellum led to the greatest reductions in black root rot incidence (59.3 and 74.1 %) and severity (72.5 and 75.2 %), as compared to un-solarized control plots. In vivo studies, this treatment dramatically increased the activity of defensive enzymes (peroxidase and chitinase) and strawberry yield (60.5 and 60.0 %, respectively), as compared to non-solarized control plots. In two field studies, the rhizosphere population of native Trichoderma spp. Developed more in solarized soils after the application of alginate pellets based on T. asperellum (86.5 and 83.6 %, respectively), compared to the non-solarized control.

This work aimed to evaluate the potential of Trichoderma asperellum organic extract and its emulsion to control cocoa black pod disease caused by Phytophthora megakarya. Organic extract was obtained after fermentation of T. asperellum and its emulsion prepared by emulsification. The in vitro antimicrobial assays of organic extract and its emulsion were evaluated and the in situ tests were carried out on detached cocoa pods. T. asperellum inhibited the mycelia growth of P. megakarya at the rates of 52% and 100%, respectively, on dual culture and the cellophane plate. This antagonist produced lytic enzymes such as cellulase, amylase, lipase and protease. The organic extract contained alkaloid, flavonoid and phenol compounds. The emulsion obtained was stable. At 100 μg · ml -1, the extract and its formulation completely inhibited the mycelial growth of P. megakarya. Similarly, when infected detached cocoa pods were sprayed with extract or emulsion, there was a significant reduction of necrosis both for healing and prevention with the latter being the most efficient. For the preventive tests, the total inhibition was recorded at 3000 μg · ml -1 and 1000 μg · ml -1, respectively, with crude organic extract and its emulsion· For curative tests, total inhibition was obtained at 4000 μg · ml -1 and 3000 μg · ml -1, respectively, for preventive and curative tests. There was a significant and positive correlation between the content of biochemical markers and the reduction of necrosis on cocoa pods after treatment with the extract or its formulation. T. asperellum organic extract emulsion could be used as an alternative in the bio- protection of cocoa black pods disease.

Penyakit akar gada pada tanaman kubis, yang disebabkan oleh jamur Plasmodiophora brassicae, merupakan ancaman serius bagi pertanian kubis. Penyakit ini menyebar dengan cepat pada suhu hangat dan kondisi pH yang asam. Limbah kotoran sapi yang dibuang oleh peternak sapi di Desa Pujon dialirkan ke dalam saluran air dan digunakan untuk irigasi pertanian, yang berpotensi mencemari lahan pertanian dan meningkatkan suhu dan keasaman tanah. Penggunaan kotoran sapi mentah sebagai bahan organik pada lahan pertanian turut mempercepat pertumbuhan jamur P. brassicae. Pengendalian biologis dengan menggunakan Trichoderma asperellum dan kapur menunjukkan potensi dalam mengurangi penyakit akar gada dan membantu dalam dekomposisi. Penelitian ini bertujuan untuk mengevaluasi efektivitas T. asperellum dan kapur dalam mendekomposisi kotoran sapi dan mengendalikan penyakit akar gada. Penelitian dilakukan di Desa Pujon Kidul, Malang, dan Laboratorium Penyakit Tumbuhan di Universitas Brawijaya selama periode lima bulan mulai dari Februari hingga Juni 2021. Penelitian terdiri dari dua tahap: pertama, dekomposisi kotoran sapi menggunakan T. asperellum dan kapur, dan kedua, aplikasi produk dekomposisi pada tanaman kubis yang terinfeksi penyakit akar gada. T. asperellum dan kapur efektif mengurangi insiden penyakit akar gada menjadi 0% dalam perlakuan T5P1 (20 hari penambahan T. asperellum pada kotoran sapi + kontrol), T4P2 (15 hari penambahan T. asperellum pada kotoran sapi + kapur), dan T5P2 (20 hari penambahan T. asperellum pada kotoran sapi + kapur), sementara pengamatan mikroskopis gejala penyakit menunjukkan penurunan hingga 10% dalam perlakuan T5P2 (20 hari penambahan T. asperellum pada kotoran sapi + kapur), menunjukkan tingkat efikasi yang tinggi pada perlakuan tersebut.

Cover crops during the off-season and multifunctional microorganisms represent strategic technologies with potential to enhance the sustainability of soybean production. This study aimed to investigate the effects of cover crop mixes and the application of multifunctional microorganisms on the economic analysis, gas exchange, yield components and grain yield of soybean plants. The experimental design followed a randomized block pattern, in a 6 x 2 factorial arrangement, with four replications. The treatments consisted of six cover crop combinations [fallow (control); millet (Pennisetum glaucum) with crotalarias (Crotalaria juncea, C. spectabilis and C. ochroleuca); millet and pigeon pea (Cajanus cajanus); millet and Urochloa ruziziensis; millet, U. ruziziensis and pigeon pea; millet and buckwheat (Fagopyrum esculentum)], with or without the application of a Trichoderma asperellum pool. The combination of millet + U. ruziziensis and millet + U. ruziziensis + pigeon pea showed the highest dry matter production and yielded the greatest nutrients content in the straw, which could provide a reduction in fertilization for the following crop. The soybean plants cultivated after millet + U. ruziziensis and millet + U. ruziziensis + pigeon pea demonstrated elevated photosynthetic rates and improved the instantaneous water-use efficiency. The application of multifunctional microorganisms led to a 16 % increase in the photosynthetic rate of the soybean plants. The highest yield was achieved by the soybean plants cultivated in areas with millet + U. ruziziensis and millet + U. ruziziensis + pigeon pea. Moreover, the application of multifunctional microorganisms contributed to increase the pod count per meter, grains per pod, mass of 100 seeds and overall soybean grain yield.

Abstract Background Chickpea plant (Cicer arietinum L.) is an important legume crop that is vulnerable to various fungal pathogens causing significant yield losses. Among them, Fusarium equiseti is a pathogen that has started to raise concern. In contrast, Trichoderma species have been explored for their ability to control such pathogens. In this study, the efficacy of a novel seed treatment formulation was explored for controlling F. equiseti in chickpea plants. The formulation was designated to enhance growth in chickpea plants as well as the ability to protect plants from infection. In addition, this formulation was tested for its effectiveness in maintaining the conidia of the antagonist in the soil after sowing. Results Applying the Trichoderma asperellum-based formulation promoted growth, as well as root and aerial biomass. In seedlings derived from treated seeds, the shoot length increased by 36.8%, and the average number of leaves also increased than the control. Following evaluation of disease severity and the foliar alteration index (FAI), a protective effect was noted, as the symptoms of Fusarium were significantly reduced in treated plants than the infected control. Re-isolation from plants infected with F. equiseti was successful in the roots (72.7%), root crown (84.5%), stem (64.4%), and even in petioles (36.1%). Conclusions Due to both direct antagonist activity and indirect growth promotion ability, the findings suggested that tested formulation can be a sustainable and eco-friendly alternative to chemical fungicides for managing F. equiseti in chickpea seeds.

This study investigated the biosynthesis, statistical optimization, characterization, and biocontrol activity of silver nanoparticles (AgNPs) produced by newly isolated Trichoderma sp. The Trichoderma asperellum strain TA-3N was identified based on the ITS gene sequence, together with its phenotypic characteristics (GenBank accession number: OM321439). The color change from light yellow to brown after the incubation period indicates AgNPs biosynthesis. The UV spectrum revealed a single peak with the maximum absorption at 453 nm, indicating that T. asperellum produces AgNPs effectively. A Rotatable Central Composite Design (RCCD) was used to optimize the biosynthesis of AgNPs using the aqueous mycelial-free filtrate of T. asperellum. The optimal conditions for maximum AgNPs biosynthesis (156.02 µg/mL) were predicted theoretically using the desirability function tool and verified experimentally. The highest biosynthetic produced AgNPs by T. asperellum reached 160.3 µg/mL using AgNO3 concentration of 2 mM/mL, initial pH level of 6, incubation time of 60 h, and biomass weight of 6 g/100 mL water. SEM and TEM imaging revealed uniform spherical shape particles that varied in size between 8.17 and 17.74 nm. The synthesized AgNPs have a Zeta potential value of −9.51 mV. FTIR analysis provided insights into the surface composition of AgNPs, identifying various functional groups such as N–H, -OH, C-H, C=O, and the amide I bond in proteins. Cytotoxicity and genotoxicity assays demonstrated that AgNPs in combination with T. asperellum can mitigate the toxic effects of Fusarium oxysporum on barley. This intervention markedly enhanced cell division rates and decreased chromosomal irregularities. The results indicate that AgNPs synthesized by T. asperellum show the potential as an eco-friendly and efficient method for controlling plant diseases. Further studies are necessary to investigate their possible use in the agricultural sector.

In order to explore the application potential of aspartic protease (Asp) of Trichoderma sp., the protease gene (asp) was cloned from Trichoderma asperellum by real-time polymerase chain reaction, and was successfully expressed in Pichia Pastoris GS115. The recombinant protease (rAsp) was isolated and purified, and its biochemical properties and its effectiveness in hydrolyzing soy protein isolate (SPI) were studied. The results showed that the protease encoded by the asp gene belonged to the aspartic protease family, and its sequence identity with other members of this family was up to 47.74%. The protease activity of rAsp in the fermentation broth obtained by induced expression in a conical flask was 25.8 U/mL. The optimal reaction pH and temperature of rAsp were 2.5 and 45 ℃, respectively, and rAsp had strong stability in the pH range of 2.0–6.0 and below 45 ℃. The activity of rAsp was promoted by Cu2+ and Mn2+ but inhibited by Fe3+, sodium dodecylsulfate (SDS) and pepstantin. The hydrolysis efficiency of SPI with rAsp was 7.7% higher than that with commercial pepsin. Moreover, the ability of rAsp to reduce the allergenicity of β-conglycinin and glycinin was 1.4 and 1.8 times greater than that of the pepsin, respectively. Therefore, rAsp has potential application in soy protein processing.

Fusarium oxysporum is the primary pathogen of blueberry root rot; furthermore, we found that Fusarium commune can also cause root rot in blueberries. Trichoderma spp. is widely used to control plant diseases. We isolated Trichoderma asperellum (TM11) from blueberry rhizosphere soil to explore its control effect and mechanism on F. oxysporum and F. commune. We found that the inhibitory effects of TM11 volatiles and broth metabolites on F. oxysporum were significant, but only F. commune volatile metabolites had a significant inhibitory effect on its growth. Twelve known antimicrobial metabolites were detected from the methanol extract of TM11 fermentation broth by HPLC-MS. TM11 lysed and coiled around the hyphae of F. oxysporum and F. commune. The pot experiment showed that TM11 had significant control effects against F. oxysporum and F. commune, and inoculation of TM11 prior to that of F. oxysporum and F. commune was more effective. The TM11, TM11 and F. oxysporum, or F. commune and distilled water treatments had different effects on the activities of superoxide dismutase, peroxidase and catalase, and the enzyme activity levels exhibited the following order: TM11 > TM11 and F. oxysporum or F. commune > distilled water. The results showed that TM11 provided effective control of blueberry root rot.

Abstract Background One of the primary corn diseases, banded leaf and sheath blight, is carried on by the pathogenic fungus Rhizoctonia solani. Efforts to control R. solani are more directed at environmentally friendly control using a biological control agent, such as Trichoderma spp. as antagonistic agents and plant growth promoter. This study aimed to identify T. asperellum isolates based on molecular characteristics and to determine the in vitro inhibition mechanism against R. solani and its role in enhancing the growth of corn seedlings. Results The HMRP7, HMRF7A, HMEDF1B, HMEDF6A, and CHM01 isolates were identified as T. asperellum with a genetic distance coefficient value of 0.000 and a very high similarity of 100%. Meanwhile, the RsHM isolate was identified as R. solani with a homology level of > 90% and genetic distance coefficient values ranging from 0.000 to 0.032. The antagonistic ability showed that the five T. asperellum isolates were able to inhibit the growth of R. solani in vitro on PDA medium with an inhibition percentage of ≥ 50%, so they were categorized as antagonist agents. T. asperellum showed the antagonistic mechanism in inhibiting the R. solani growth through the action of parasitism. The five T. asperellum isolates tested on corn seedlings showed a significantly high difference from the control treatment on the observational variables of maximum growth potential (MGP), growth rate (GtR), growth simultaneity (GS), vigor index (VI), germination rate (GR), and median germination time (T50). Conclusion CHM01 isolate showed better potential than other isolates in inhibiting the growth of R. solani in vitro on PDA medium with a parasitism mechanism and enhancing the growth of corn seedlings.

Damping-off disease, caused by Pythium aphanidermatum, significantly impacts tomato production. This study explored the potential of a two-pronged approach for enhanced biocontrol: combining two antagonistic Trichoderma asperellum strains (CB-Pin-01 and NST-009) with a plant immune stimulant (CaCO3). Laboratory assays demonstrated strong individual efficacy of both Trichoderma strains against P. aphanidermatum, with significant growth inhibition and overgrowth capabilities. Importantly, scanning electron microscopy confirmed their compatibility. Greenhouse experiments revealed that the combined application of Trichoderma strains and CaCO3 achieved the most significant reduction in disease incidence (17.78%) compared to the control (66.55%). Furthermore, this treatment resulted in 100% root colonization by Trichoderma and the highest population density in the soil (6.17 × 107 CFU g−1), suggesting the immune stimulant’s role in promoting beneficial microbe establishment. These findings highlight the potential of this combined strategy as a sustainable and effective approach for managing damping-off disease in tomatoes.

This study evaluated the initial development of paricá (Schizolobium parahyba var. amazonicum) seedlings under different nitrogen rates with the application of Trichoderma spp., using a randomized complete block design in a 4x5 factorial scheme (strains and rates) with seven replications. The evaluated traits were plant height, stem diameter, leaf and stem fresh weights, leaf and stem dry weights, and aerial part dry and fresh weights. Trichoderma spp. strains did not satisfactorily promote paricá seedlings (Schizolobium parahyba var. amazonicum) under high nitrogen rates. However, the Trichoderma harzianum IBLF 006 WP strain was efficient only under low nitrogen availability.

Abstract The biological products use as a disease control alternative has been studied to reduce the impacts to the environment, men and animals, showing satisfactory results in postharvest. This study aimed to evaluate the biological agents effect in the control of Colletotrichum spp. and on postharvest quality of yellow passion fruit. The treatments were Trichoderma asperellum and Saccharomyces cerevisiae species at concentrations of of 0,5; 1,0; 1,5; 2,0 (g. L-1); Mancozeb fungicide (Dithane® 2 g i.a. L-1 water) and control (sterile distilled water). The fruits were immersed for 2 min in each treatment and then were drought. Five replications of three fruits were used to anthracnose severity analysis and yellow passion fruit physical-chemical quality in. The fruit inoculation was made with the deposition of Colletotrichum spp. on the surface of the fruit previously treated using holes which were made with the aid of a flamed perforator. In the research was evaluated: pH, total soluble solids and titratable acidity. Biological treatments reduced the anthracnose severity in yellow passion fruit. Fruit Post-harvest quality was not influenced by the biological control application. It is a viable alternative to postharvest management of anthracnose on yellow passion fruit under the studied conditions.

Mannan, a highly abundant and cost-effective natural resource, holds great potential for the generation of high-value compounds such as bioactive polysaccharides and biofuels. In this study, we successfully enhanced the expression of constructed GH5 β-mannanase (TaMan5) from Trichoderma asperellum ND-1 by employing propeptide in Pichia pastoris. By replacing the α-factor with propeptide (MGNRALNSMKFFKSQALALLAATSAVA), TaMan5 activity was significantly increased from 67.5 to 91.7 U/mL. It retained higher activity in the presence of 20% ethanol and 15% NaCl. When incubated with a high concentration of mannotriose or mannotetraose, the transglycosylation action of TaMan5 can be detected, yielding the corresponding production of mannotetraose or mannooligosaccharides. Moreover, the unique mechanism whereby TaMan5 catalyzes the degradation of mannan into mannobiose involves the transglycosylation of mannose to mannotriose or mannotetraose as a substrate to produce a mannotetraose or mannopentose intermediate, respectively. Additionally, the production of soluble sugars from lignocellulose is a crucial step in bioethanol development, and it is noteworthy that TaMan5 could synergistically yield fermentable sugars from corn stover and bagasse. These findings offered valuable insights and strategies for enhancing β-mannanase expression and efficient conversion of lignocellulosic biomass, providing cost-effective and sustainable approaches for high-value biomolecule and biofuel production.

Most Trichoderma species are beneficial fungi that promote plant growth and resistance, while Fusarium genera cause several crop damages. During the plant-fungi interaction there is a competition for sugars in both lifestyles. Here we analyzed the plant growth promotion and biocontrol activity of T. asperellum against F. verticillioides and the effect of both fungi on the expression of the maize diffusional sugar transporters, the SWEETs. The biocontrol activity was done in two ways, the first was by observing the growth capacity of both fungus in a dual culture. The second one by analyzing the infection symptoms, the chlorophyl content and the transcript levels of defense genes determined by qPCR in plants with different developmental stages primed with T. asperellum conidia and challenged with F. verticillioides. In a dual culture, T. asperellum showed antagonist activity against F. verticillioides. In the primed plants a delay in the infection disease was observed, they sustained chlorophyll content even after the infection, and displayed upregulated defense-related genes. Additionally, the T. asperellum primed plants had longer stems than the nonprimed plants. SWEETs transcript levels were analyzed by qPCR in plants primed with either fungus. Both fungi affect the transcript levels of several maize sugar transporters differently. T. asperellum increases the expression of six SWEETs on leaves and two at the roots and causes a higher exudation of sucrose, glucose, and fructose at the roots. On the contrary, F. verticillioides reduces the expression of the SWEETs on the leaves, and more severely when a more aggressive strain is in the plant. Our results suggest that the plant is able to recognize the lifestyle of the fungi and respond accordingly by changing the expression of several genes, including the SWEETs, to establish a new sugar flux.

Sustainable plant production requires less use of synthetic chemicals in plant nutrition and protection. Microbial products are among the most promising substitutes for chemicals. With the increasing popularity and availability of such products, it has become obligatory to use different microbes together. The effect of this has been tested in several studies, but their results have sometimes been contradictory depending on the microbial strains tested and the mode of application. We tested the effect of two commercially available antagonists and Funneliformis mosseae alone and in combination on tomato. Mycorrhizal treatment increased plant growth and yield, both alone and combined with the antagonists; however, mycorrhizal root colonization was not influenced by the antagonist. This treatment also led to a slight decrease in the occurrence of Trichoderma spp. on tomato roots but did not impede the colonization of roots by the applied Trichoderma strain. Our result confirmed that Trichoderma asperellum (T34) and Streptomyces griseoviridis (K61) can be safely combined with arbuscular mycorrhizal fungi (AMF), namely with F. mosseae.

Strawberry root rot caused by Neopestalotiopsis clavispora is one of the main diseases of strawberries and significantly impacts the yield and quality of strawberry fruit. Currently, the only accessible control methods are fungicide sprays, which could have an adverse effect on the consumers of the strawberries. Biological control is becoming an alternative method for the control of plant diseases to replace or decrease the application of traditional synthetic chemical fungicides. Trichoderma spp. are frequently used as biological agents to prevent root rot in strawberries. In order to provide highly effective biocontrol resources for controlling strawberry root rot caused by Neopestalotiopsis clavispora, the biocontrol mechanism, the control effects of T. asperellum CMT10 against strawberry root rot, and the growth-promoting effects on strawberry seedlings were investigated using plate culture, microscopy observation, and root drenching methods. The results showed that CMT10 had obvious competitive, antimycotic, and hyperparasitic effects on N. clavispora CMGF3. The CMT10 could quickly occupy nutritional space, and the inhibition rate of CMT10 against CMGF3 was 65.49% 7 d after co-culture. The inhibition rates of volatile metabolites and fermentation metabolites produced by CMT10 were 79.67% and 69.84% against CMGF3, respectively. The mycelium of CMT10 can act as a hyperparasite by contacting, winding, and penetrating the hyphae of CMGF3. Pot experiment showed that the biocontrol efficiency of CMT10 on strawberry root rot caused by Neopestalotiopsis clavispora was 63.09%. CMT10 promoted strawberry growth, plant height, root length, total fresh weight, root fresh weight, stem fresh weight, and root dry weight by 20.09%, 22.39%, 87.11%, 101.58%, 79.82%, and 72.33%, respectively. Overall, this study showed the ability of T. asperellum CMT10 to control strawberry root rot and its potential to be developed as a novel biocontrol agent to replace chemical fungicides for eco-friendly and sustainable agriculture.

Abstract Background Global warming caused by greenhouse gas emissions affects sustainable human development. Agricultural practices are important source of greenhouse gases (GHG). Nitrous oxide (N2O) contributes greatly to farming GHG. It is important to find a potential and practical biological technique that mitigate N2O emissions in an environment friendly way. Methods N2O-inhibiting fungi were isolated and identified in the lab. The fungi were added into the soil and placed in the incubator and interval gas sampling was analyzed by gas chromatograph. Results Fungus coding Z17 was identified molecularly with the same evolutionary branch on the phylogenetic tree with Trichoderma asperellum by BLAST comparison on NCBI GenBank. In the lab simulation, the N2O emission flux was decreased by 28.18–47.16% by inoculating Trichoderma asperellum with 106 cfu·g−1, 5 × 106 cfu·g−1 and 107 cfu·g−1 fungal spores in the soil compared to the control. Conclusions The N2O-inhibiting fungus Z17 was identified as Trichoderma asperellum, capable of suppressing N2O emissions from soil with at least 106 CFU·g−1 soil. The best N2O-inhibiting effect was on day 9 of inoculation into soil because most of the fungal numbers were present in soil. Graphical Abstract

Abstract Background Yellow (stripe) rust, caused by Puccinia striiformis f. sp. tritici (Pst), is an economic disease of wheat. Growth-promoting fungi (GPF) such as Trichoderma asperellum and Penicillium simplicissimum have been investigated for their potential to control yellow rust and their involvement in gene expression of four PR proteins for all-stage resistance. Results Wheat plants (cv. Sids-12) treated individually with each of the biocontrol agents, P. simplicissimum and T. asperellum, at 24 and 48 hpi showed a resistance response (infection type = 2) to yellow rust, compared to the non-treated plants, which showed highly susceptible response (infection type = 9). Both biocontrol agents induced resistance against yellow rust on wheat plants, exhibiting a moderate resistance (10 MR) and reduced the colony size of Pst (0.6 mm2). Moreover, P. simplicissimum and T. asperellum increased (P ≤ 0.05) the grain yields of wheat plants infected with Pst. Scanning electron microscope (SEM) of yellow rust infected wheat leaves treated with P. simplicissimum and T. asperellum at 24 and 48 hpi showed hyperparasitism on Pst urediniospores and inhibition of the spore germination. Expressions of pathogenesis-related (PR) protein genes, PR1, PR2, PR3 and PR4 were higher in wheat plants treated with both biocontrol agents than the non-treated checks. Conclusion P. simplicissimum and T. asperellum exhibited biocontrol potential against yellow rust disease caused by P. striiformis f. sp. tritici (Pst) on wheat plants. It was found that wheat defence mechanism against Pst was activated by a high expression of PR protein genes induced by both biocontrol agents.

In this study, an acidophilic GH5 β-mannanase (TaMan5) from Trichoderma asperellum ND-1 was efficiently expressed in Pichia pastoris (a 2.0-fold increase, 67.5 ± 1.95 U/mL). TaMan5 displayed the highest specificity toward locust bean gum (Km = 1.34 mg/mL, Vmax = 749.14 μmol/min/mg) at pH 4.0 and 65°C. Furthermore, TaMan5 displayed remarkable tolerance to acidic environments, retaining over 80% of its original activity at pH 3.0–5.0. The activity of TaMan5 was remarkably decreased by Cu2+, Mn2+, and SDS, while Fe2+/Fe3+ improved the enzyme activity. A thin-layer chromatography (TLC) analysis of the action model showed that TaMan5 could rapidly degrade mannan/MOS into mannobiose without mannose via hydrolysis action as well as transglycosylation. Site-directed mutagenesis results suggested that Glu205, Glu313, and Asp357 of TaMan5 are crucial catalytic residues, with Asp152 playing an auxiliary function. Additionally, TaMan5 and commercial α-galactosidase displayed a remarkable synergistic effect on the degradation of galactomannans. This study provided a novel β-mannanase with ideal characteristics and can be considered a potential candidate for the production of bioactive polysaccharide mannobiose.

Abstract Background Trichoderma asperellum (Ascomycota: Hypocreaceae) is a globally recognized soil fungus due to its broad-spectrum antimicrobial and plant growth-promoting properties. To increase the availability of local strains, soil samples from a dragon fruit farm in Villanueva (Misamis Oriental, Philippines) were collected and baited using the insect-baiting technique. Results Using the baiting technique, T. asperellum strain, RMCK01, was isolated. The strain was characterized using morphological and molecular data and its biological control potential was tested using different conidial concentrations on the last larval instar of the super worm, Zophobas morio (Fabricius, 1776) (Coleoptera: Tenebrionidae). The ITS1 sequences of T. asperellum RMCK01 were 100% identical to the sequences of other T. asperellum isolates reported from Vietnam, India, Thailand, and China. In addition, T. asperellum RMCK01 was particularly efficient against Z. morio larvae. On day 21, 88.87%, 88.07%, and 86.73% of insects died when treated with a suspension containing 6 × 108, 3 × 109, and 2.68 × 107 conidia/ml, respectively. Conclusion The study highlights the potential of this fungal isolate as a biocontrol agent against insect pests.

Abstract Background The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is an invasive alien species in Indonesia. This study aimed to assess the impact of its invasion in Indonesia by measuring the damaged area caused by the pest in maize fields located across East Java and simultaneously determine whether S. frugiperda outcompetes the native Asian armyworm Spodoptera litura (Fabricius). Secondly, the virulence of 14 entomopathogenic fungus (EPF) isolates against S. frugiperda larvae was evaluated in an effort to find effective biocontrol agent candidates. Results The damaged area caused by S. frugiperda was generally higher than that caused by S. litura during the survey period from August 2019 to December 2021. It indicated that S. frugiperda may have dominated the native armyworm and become the primary key pest of maize in Indonesia. Based on a single-concentration assay (106 conidia ml−1), the tested EPF isolates displayed varying degrees of virulence against S. frugiperda larvae, causing larval mortality of 3.5 to 71% at 10-day post-treatment, with the highest mortality rates provided by Beauveria bassiana sensu lato and Trichoderma asperellum sensu lato. At a concentration of 108 conidia ml−1, B. bassiana s.l. and T. asperellum s.l. elicited high larval mortality of 76 and 81%, respectively, at 10-day post-treatment. Nevertheless, the probit analysis based on a concentration–response assay revealed that T. asperellum s.l. had lower LC50 and LC90 values than B. bassiana s.l. Conclusions The attack and invasion of S. frugiperda seem to be a continual threat to the maize agro-ecosystem in Indonesia. As a consequence, Indonesia should mitigate and be well-prepared for future outbreaks of S. frugiperda. Indigenous EPF isolates used in this study may act as promising biocontrol agents of S. frugiperda, especially T. asperellum s.l. This study also serves as the first report documenting the direct lethality of Trichoderma fungus on S. frugiperda larvae.

Wheat blast caused by Magnaporthe oryzae Pathotype triticum (MoT) becomes one of the common nuisances to the expansion of wheat production in Bangladesh That's why to find out a suitable control measure, fourteen chemicals and five Trichoderma isolates were evaluated against MoT in in-vitro condition during the cropping seasons of 2019-2020. In in-vitro conditions the efficacies of fungicides and biological agents i.e. Trichoderma isolates were evaluated by the measurement of percent inhibition of radial mycelial growth of MoT. The highest percentage of radial mycelial growth inhibition (100%) of MoT was obtained by Mancozeb 63% + Carbendazim 12% (Companion), Carbendazim (Autostin 50 WDG), Pyrochlostrobin 5% + Metiram 55% (Cabrio Top), Tebuconazol (Folicur EW 250) and Hexaconazol (Contaf 5EC) fungicides at low concentration (0.0125%) compared to other fungicides at 10 days after inoculation (DAI). Out of five Trichoderma asperellum isolates used in this study, the maximum percent of radial mycelial growth inhibition (58.24%) of MoT was achieved by Trichoderma isolate PMILTE-N51 (58.24%) followed by isolate PMILTE- N41 (53.85%) in the in-vitro dual culture assay at 7 DAI. In case of pot experiment, seven fungicides (Nativo 75WG, Companion, Autostin, Indofil M45, Supreme 250EW, Contaf, Amiscore 32.5 SC) at 0.1% & 0.3% and bioagents (Trichoderma asperellum, PMILTE-N51-suspension, PMILTE-N51- formulation & Trichoderma herzianum based IPM LAB- Formulation) were used to evaluate their efficacy in controlling wheat blast disease and their effects on different yield contributing parameters of wheat. Minimum percentage of blast incidence and severity were recorded for Supreme 250 EW @ 0.3% (14.88% and 2.21%) treated plants followed by PMILTE-N51 suspension compared to control (85.06% and 90.2%). In case of different yield contributing parameters viz. maximum number of ear/pot (13.67), number of total and healthy spikelets/ear (55.33 and 52.33), number of total and healthy grains/ear (50.33 and 45.67 ) were achieved by Amiscore 32.5 SC @ 0.3% followed by Supreme 250EW @ 0.3% and a maximum weight of 1000 grains/pot (55 gm) was recorded for PMILTE-N51-suspension, whereas controlled plants showed the lowest value for all the yield contributing parameters among the treatments used in this experiment. These results indicate that Amiscore 32.5 SC and Supreme 250 EW (@ 0.3%) showed the best results as chemical control approaches, where Trichoderma (PMILTE-N51-suspension) can be a better non-chemical management approach against wheat blast in Bangladesh. [Fundam Appl Agric 2022; 7(1.000): 47-59]

Plant-associated microbes play crucial roles in plant health and promote growth under stress. Tomato (Solanum lycopersicum) is one of the strategic crops grown throughout Egypt and is a widely grown vegetable worldwide. However, plant disease severely affects tomato production. The post-harvest disease (Fusarium wilt disease) affects food security globally, especially in the tomato fields. Thus, an alternative effective and economical biological treatment to the disease was recently established using Trichoderma asperellum. However, the role of rhizosphere microbiota in the resistance of tomato plants against soil-borne Fusarium wilt disease (FWD) remains unclear. In the current study, a dual culture assay of T. asperellum against various phytopathogens (e.g., Fusarium oxysporum, F. solani, Alternaria alternata, Rhizoctonia solani, and F. graminerarum) was performed in vitro. Interestingly, T. asperellum exhibited the highest mycelial inhibition rate (53.24%) against F. oxysporum. In addition, 30% free cell filtrate of T. asperellum inhibited F. oxysporum by 59.39%. Various underlying mechanisms were studied to explore the antifungal activity against F. oxysporum, such as chitinase activity, analysis of bioactive compounds by gas chromatography–mass spectrometry (GC–MS), and assessment of fungal secondary metabolites against F. oxysporum mycotoxins in tomato fruits. Additionally, the plant growth-promoting traits of T. asperellum were studied (e.g., IAA production, Phosphate solubilization), and the impact on tomato seeds germination. Scanning electron microscopy, plant root sections, and confocal microscopy were used to show the mobility of the fungal endophyte activity to promote tomato root growth compared with untreated tomato root. T. asperellum enhanced the growth of tomato seeds and controlled the wilt disease caused by the phytopathogen F. oxysporum by enhancing the number of leaves as well as shoot and root length (cm) and fresh and dry weights (g). Furthermore, Trichoderma extract protects tomato fruits from post-harvest infection by F. oxysporum. Taking together, T. asperellum represents a safe and effective controlling agent against Fusarium infection of tomato plants.

Trichoderma asperellum PK1J2 is a promising cellulase-producing fungus isolated from a palm empty fruit bunch in Riau, Indonesia. Presented here is the genome assembly of T. asperellum PK1J2. The whole genome of the fungi was sequenced using Illumina NovaSeq PE150. The genome assembly was performed using SOAPdenovo, SPAdes, and Abyss software, and the assembly results of the three types of software were integrated with CISA software. T. asperellum PK1J2 has 6,835 protein-coding genes with a length of 9,233,597 bp. The final genome assembly was approximately 36 Mbp with a GC content of 48.45%. This whole genome shotgun project has been deposited at DDBJ/ENA/GenBank under accession JAGJIK000000000.

Managing organic agricultural wastes is a challenge in today’s modern agriculture, where the production of different agricultural goods leads to the generation of large amounts of waste, for example, olive pomace and vine shoot in Mediterranean Europe. The discovery of a cost-effective and environment-friendly way to valorize such types of waste in Mediterranean Europe is encouraged by the European Union regulation. As an opportunity, organic agricultural waste could be used as culture media for solid-state fermentation (SSF) for fungal strains. This methodology represents a great opportunity to produce secondary metabolites like 6-pentyl-alpha-pyrone (6-PP), a lactone compound with antifungal properties against phytopathogens, produced by Trichoderma spp. Therefore, to reach adequate yields of 6-PP, lytic enzymes, and spores, optimization using specific agricultural cheap local wastes from Southeastern France is in order. The present study was designed to show the applicability of an experimental admixture design to find the optimal formulation that favors the production of 6-PP. To conclude, the optimized formulation of 6-PP production by Trichoderma under SSF contains 18% wheat bran, 23% potato flakes, 20% olive pomace, 14% olive oil, 24% oatmeal, and 40% vine shoots.

Trichoderma, a well-known and extensively studied fungal genus, has gained significant attention for its remarkable antagonistic abilities against a wide range of plant pathogens. In this study, a total of 108 Trichoderma isolates were screened through in vitro dual antagonistic assays and culture filtrate inhibition against Fusarium graminearum. Of these, the YNQJ1002 displayed noteworthy inhibitory activities along with thermal stability. To validate the metabolic differences between YNQJ1002 and GZLX3001 (with strong and weak antagonism, respectively), UPLC-TOF-MS/MS mass spectrometry was employed to analyze and compare the metabolite profiles. We identified 12 significantly up-regulated metabolites in YNQJ1002, which include compounds like Trigoneoside, Torvoside, trans,trans-hepta-2,4,6-trienoic acid, and Chamazulene. These metabolites are known for their antimicrobial properties or signaling roles as components of cell membranes. Enriched KEGG analysis revealed a significant enrichment in sphingolipid metabolism and linoleic acid metabolism, as well as autophagy. The results demonstrated that YNQJ1002’s abundance of antimicrobial substances, resulting from specific metabolic pathways, enhanced its superior antagonistic activity against F. graminearum. Finally, YNQJ1002 was identified using the ITS, tef1-1α, and rpb2 regions, with MIST system sequence matching confirming its classification within the species. Overall, we have obtained a novel strain, T. asperellum YNQJ1002, which is rich in metabolites and shows potential antagonistic activity against F. graminearum. This study has opened promising prospects for the development of innovative Trichoderma-derived antifungal compounds, featuring a unique mechanism against pathogens.

Stem-end rot disease has been causing damage to the production of pomelos in Vietnam. The cur-rent study aimed to (i) isolate fungal pathogens causing pomelo stem-end rot disease (PSERD) and (ii) discover Trichoderma spp. that had an antagonistic ability against pathogens under in vitro conditions. Fungi causing PSERD were isolated from pomelo fruits with symptoms of stem-end rot disease and collected from pomelo farms in Ben Tre province, Vietnam. Moreover, 50 fungal strains of Trichoderma spp. also originated from soils of these pomelo farms in Ben Tre province and were dual-tested with the fungal pathogen on the PDA medium. The results demonstrated that 11 pathogenic fungi causing PSERD were isolated from the fruit and showed mycelial growth of roughly 5.33–8.77 cm diameter at 72 h after inoculation. The two fungi that exhibited the fast-est growth, namely, S-P06 and S-P07, were selected. ITS sequencing of the S-P06 and S-P07 fungi resulted in Lasiodiplodia theobromae. All the 50 Trichoderma spp. strains were allowed to antago-nize against the S-P06 and S-P07 strains under in vitro conditions. The greatest antagonistic effi-ciency was found in Trichoderma spp. T-SP19 at 85.4–86.2% and T-SP32 at 84.7–85.4%. The two antagonists were identified as Trichoderma asperellum T-SP19 and T-SP32. The selected strains of Trichoderma asperellum were potent as a biological control for fruit plants.

Penyakit budok yang disebabkan oleh Synchytrium pogostemonis merupakan masalah utama pada budi daya nilam di Provinsi Aceh (Pogostemon cablin). Tujuan penelitian ini ialah untuk mendapatkan cara pengendalian S. pogostemonis dengan menggabungkan kompos limbah ampas nilam (LAN), mikrob antagonis, dan minyak atsiri sebagai anticendawan. Penelitian dibagi menjadi dua tahap. Tahap pertama ialah pengujian efek penambahan kompos LAN terhadap pertumbuhan tanaman nilam. Tiga spesies mikrob antagonis, yaitu Trichoderma harzianum (ThC6), T. asperellum (TaC1), dan Pseudomonas aureginosa (AJ14) diinokulasikan pada LAN sebagai biodekomposer sekaligus agens antagonis baik secara tunggal maupun kombinasi. Kompos LAN diaplikasikan pada medium tanam sebelum penanaman setek nilam dengan dua taraf rasio tanah dan kompos LAN (1:1 dan 1:2). Parameter yang diamati ialah pertumbuhan tanaman nilam meliputi tinggi tanaman dan jumlah daun. Pada tahap kedua diamati pengaruh kombinasi kompos LAN dan minyak atsiri terhadap keparahan penyakit budok. Minyak atsiri yang digunakan adalah minyak atsiri serai wangi dan nilam dengan konsentrasi 0.5% dan 1%. Hasil penelitian menunjukkan bahwa penambahan mikrob antagonis baik secara tunggal maupun kombinasi pada LAN dapat meningkatkan tinggi batang dan jumlah daun tanaman nilam masing-masing sebesar 22% dan 38%. Keparahan penyakit terendah ditunjukkan oleh perlakuan kombinasi kompos LAN dan 1% minyak atsiri serai wangi, yaitu sebesar 10%.

Acidophilic β-xylanases have attracted considerable attention due to their excellent activity under extreme acidic environments and potential industrial utilizations. In this study, a novel β-xylanase gene (Xyl11) of glycoside hydrolase family 11, was cloned from Trichoderma asperellum ND-1 and efficiently expressed in Pichia pastoris (a 2.0-fold increase). Xyl11 displayed a maximum activity of 121.99 U/ml at pH 3.0 and 50°C, and exhibited strict substrate specificity toward beechwood xylan (Km = 9.06 mg/ml, Vmax = 608.65 μmol/min/mg). The Xyl11 retained over 80% activity at pH 2.0–5.0 after pretreatment at 4°C for 1 h. Analysis of the hydrolytic pattern revealed that Xyl11 could rapidly convert xylan to xylobiose via hydrolysis activity as well as transglycosylation. Moreover, the results of site-directed mutagenesis suggested that the Xyl11 residues, Glu127, Glu164, and Glu216, are essential catalytic sites, with Asp138 having an auxiliary function. Additionally, a high degree of synergy (15.02) was observed when Xyl11 was used in association with commercial β-xylosidase. This study provided a novel acidophilic β-xylanase that exhibits excellent characteristics and can, therefore, be considered a suitable candidate for extensive applications, especially in food and animal feed industries.

This study evaluated the effect of two plant materials (Panicum maximum and Tridax procumbens) composted with cow dung, with or without Trichoderma asperellum inoculation on soil properties and yield of celosia in 2014 and 2015. The treatments were tridax-based compost without Trichoderma (TBC), tridax-based compost with Trichoderma (TTBC), panicum-based compost without Trichoderma (PBC), panicum-based compost with Trichoderma (TPBC) and control (no compost). All compost were applied at 240 kgN/ha. The design was RCBD with three replicates and data analyzed using ANOVA at α0.05. Results showed that compost enhanced growth, nutrient uptake and yield of celosia. In 2014, highest fresh weight (57.09 t/ha) was obtained from plant treated with TPBC, which compared favourably with TTBC TTBC (57.00 t/ha) but significantly higher than TBC (43.85 t/ha) and PBC (47.32 t/ha) while control gave the least significant value (20 t/ha). A similar trend was obtained in 2015. This infers that plants that received inoculated compost gave better yield compared to uninoculated compost. Post-cropping soil chemical analysis revealed that compost improved soil N, P, K and organic C. This shows that Trichoderma inoculated compost could be better than the uninoculated compost for celosia production in an Alfisol.

Trichoderma spp. are versatile probiotic fungi that are known to promote plant growth and disease resistance under biotic stress, abiotic stress, or both. They are often used to prevent plant diseases caused by pathogens, act as biofertilizers, and are used in mycoremediation. In our previous study, Trichoderma asperellum strain SM-12F1 was isolated from soils contaminated with arsenic (As), adjacent to a realgar mine. SM-12F1 promoted plant growth and was useful for biocontrol and bioremediation. However, the genomic sequence of this strain was not characterized. This study aimed to generate high-quality genome resources for T. asperellum SM-12F1, and to determine the genomic basis of mechanisms behind plant growth promotion, biocontrol, and bioremediation of As in soil. Genome data of this fungus will provide perspectives on the molecular basis underlying biocontrol activity and mycoremediation.[Figure: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Brazil has many important native species, which includes Schizolobium parahyba var. amazonicum (Huber ex Ducke) Barneby, however, the growth of this species is inhibited in soils with low boron contents, or excess boron, which causes phytotoxicity. Applications of strains of Trichoderma spp. increase the plants' tolerance to abiotic stresses, including tolerance to low or excess levels of nutrients such as boron (B). Thus, the objective of this work was to evaluate plant development and the effect of strains of Trichoderma spp. in Schizolobium parahyba var. amazonicum seedlings grown under different boron rates. A randomized block experimental design was used, with a 4×5 factorial scheme, consisting of 4 strains of Trichoderma spp. and 5 B rates (0, 0.5, 1.0, 1.5, and 2.0 mg dm-3), with seven replications, in greenhouse. Plant height, stem diameter, fresh and dry mass, leaf, stem and total were evaluated at 120 days after emergence. The stains of Trichoderma spp. and B rates presented significant interaction for all evaluated variables, decreasing phytotoxicity. The strain IBLF006WP (T. harzianum) showed a higher capacity of increasing the plants' tolerance to boron, followed by URM5911 (T. asperellum). However, the beneficial effect of increasing this tolerance with the application of these strains is only feasible for soils with high contents of this micronutrient in the soil.

Cadmium (Cd) is one of the most toxic heavy metal pollutants known to accumulate in the soil environment and pose a threat to public health. The tolerance of endophytic fungi isolated from Kosteletzkya virginica to cadmium was determined by the Oxford cup method. Trichoderma asperellum displayed the highest tolerance to cadmium on potato dextrose agar (PDA) medium and was selected subsequently for inoculating maize seedlings. Green fluorescein protein (GFP) labeled T. asperellum colonised in the roots of maize seedlings. Under Cd stress, the plant height, dry weight, fresh weight, root length, chlorophyll content and antioxidant enzyme activity of maize seedlings inoculated with T. asperellum were significantly higher than those of the control. T. asperellum also contributed to the up-regulated expression of some key genes in roots and leaves of maize seedlings, involving nitrogen absorption and metabolism to promote maize growth and Cd detoxification, accumulation, distribution, etc. for Cd transport to increase resistance to Cd.

Abstract Background Environmental pollution is one of the major problems that the world is facing today. Several approaches have been taken, from physical and chemical methods to biotechnological strategies (e.g. the use of oxidoreductases). Oxidative enzymes from microorganisms offer eco-friendly, cost–effective processes amenable to biotechnological applications, such as in industrial dye decolorization. The aim of this study was to screen marine-derived fungal strains isolated from three coastal areas in Tunisia to identify laccase-like activities, and to produce and characterize active cell-free supernatants of interest for dye decolorization. Results Following the screening of 20 fungal strains isolated from the harbors of Sfax and Monastir (Tunisia), five strains were identified that displayed laccase-like activities. Molecular-based taxonomic approaches identified these strains as belonging to the species Trichoderma asperellum, Stemphylium lucomagnoense and Aspergillus nidulans. Among these five isolates, one T. asperellum strain (T. asperellum 1) gave the highest level of secreted oxidative activities, and so was chosen for further studies. Optimization of the growth medium for liquid cultures was first undertaken to improve the level of laccase-like activity in culture supernatants. Finally, the culture supernatant of T. asperellum 1 decolorized different synthetic dyes belonging to diverse dye families, in the presence or absence of 1-hydroxybenzotriazole (HBT) as a mediator. Conclusions The optimal growth conditions to produce laccase-like active cell-free supernatants from T. asperellum 1 were 1.8 mM CuSO4 as an inducer, 1% NaCl to mimic a seawater environment and 3% sucrose as a carbon source. The culture supernatant of T. asperellum 1 effectively decolorized different synthetic dyes belonging to diverse chemical classes, and the presence of HBT as a mediator improved the decolorization process.

Abstract This study aimed to evaluate the efficiency of Enterobacter cloacae PS14 and Trichoderma asperellum T34 in the control of potato wilt, caused by Ralstonia solanacearum (Smith), under greenhouse and field conditions. In vitro, the endophyte E. cloacae PS14 caused the highest reduction of the pathogen growth among 7 screened bacteria. It produced an inhibition zone as 16.9 mm compared to a specific antibiotic (20.0 mm). E. cloacae PS14 was selected as an effective antagonistic bacterium to be compared to T. asperellum strain T34 for reduction of the disease as well as increasing the crop yield of potato plants. Both E. cloacae and T. asperellum reduced the disease severity up to 10.7–26.5%, respectively, under greenhouse and up to 26.6–36.6%, respectively, under field conditions. The results approved that both E. cloacae and T. asperellum increased the yield of the crop by 20.44–40.96%, respectively. Their mode of action was indicated by suppression of the pathogen as well as induction of plant systemic resistance. The induction of systemic resistance was confirmed by increasing the total phenol and salicylic acid contents as well as increasing the activities of peroxidase, lipoxygenase, and polyphenol oxidase in potato plants than the healthy or only infected plants. Production of siderophore, indole-3-acetic acid (0.577–0.884 μM), hydrogen cyanide (2.34–3.61 μg/ml), and salicylic acid (0.436–1.488 μg/ml) was confirmed by E. cloacae PS14 and T. asperellum T34, respectively, in vitro. The study recommends the new strain E. cloacae PS14, as new endophytic effective bacteria, in the control of R. solanacearum causing the potato wilt disease.

Abstract Environmental conditions affect biocontrol agents in a field, being appropriate formulations an alternative to overcome this problem. Formulations based on Trichoderma asperellum TV190 were prepared by emulsified mineral or vegetable oils, which protected spores from ultraviolet radiation, showing greater viability of 37–43% (mineral) and 56–63% (vegetable) than the control (8–12%). These formulations improved an antagonism of T. asperellum on Rhizoctonia solani under greenhouse conditions, reducing infected corn seedlings by 72% (mineral) and 59% (vegetable). Necrotic spot size was reduced by 90.04% (mineral) and 87.29% (vegetable). A granular formulation, prepared with degreased corn germ and T. asperellum spores, protected the corn seedlings from R. solani under greenhouse conditions, with 73% reduction of infected plants and 93% reduction of necrotic spot size. Both granular and liquid formulations were able to improve T. asperellum antagonism, suggesting that these formulations could be included in agricultural pest control strategies.

As excellent biocontrol agents and plant growth promoters, Trichoderma species are agriculturally important. Trichoderma spp. cultures can be produced using solid-state or submerged cultivation, the latter being much less labor intensive and easier to control and automate. The aim of the study was to investigate the ability to increase the shelf-life of T. asperellum cells by optimizing cultivation media and upscaling the submerged cultivation process. Four different cultivation media were used with or without the addition of Tween 80 and stored with or without incorporation into peat, and viability, expressed as CFU/g, was assessed during one year of storage in an industrial warehouse. The addition of Tween 80 had a positive effect on the biomass yield. The culture medium played a major role in the ability of the mycelium to produce spores, which in turn influenced the amount of CFU. This effect was less pronounced when the biomass was mixed with peat prior to storage. A procedure that increases the number of CFU in a peat-based product formulation is recommended, namely, incubation of the mixture at 30 °C for 10 days prior to storage at 15 °C over an extended period of time.

BackgroundBiocontrol strategies are the best possible and eco-friendly solution to develop resistance against O furnacalis and improve the maize yield. However, the knowledge about underlying molecular mechanisms, metabolic shifts, and hormonal signaling is limited.MethodsHere, we used an axenic and a consortium of entomopathogenic Beauveria bassiana OFDH1-5 and a pathogen-antagonistic Trichoderma asperellum GDFS1009 in maize and observed that consortium applications resulted in higher chlorophyll contents and antioxidants activities [superoxide dismutase (SOD), peroxidase (POD), proline, protease, and polyphenol oxidase (PPO)] with a decrease in O. furnacalis survival. We performed a comprehensive transcriptome and an untargeted metabolome profiling for the first time at a vegetative stage in fungal inoculated maize leaves at 0-, 12-, 24-, 48-, and 72-h post insect infestation.ResultsThe consortium of B. bassiana and T. asperellum leads to 80–95% of O. furnacalis mortality. A total of 13,156 differentially expressed genes were used for weighted gene coexpression network analysis. We identified the six significant modules containing thirteen candidate genes [protein kinase (GRMZM2G025459), acyl-CoA dehydrogenase (GRMZM5G864319), thioredoxin gene (GRMZM2G091481), glutathione S-transferase (GRMZM2G116273), patatin-like phospholipase gene (GRMZM2G154523), cytochrome P450 (GRMZM2G139874), protease inhibitor (GRMZM2G004466), (AC233926.1_FG002), chitinase (GRMZM2G453805), defensin (GRMZM2G392863), peroxidase (GRMZM2G144153), GDSL- like lipase (AC212068.4_FG005), and Beta-glucosidase (GRMZM2G031660)], which are not previously reported that are highly correlated with Jasmonic acid - Ethylene (JA-ET) signaling pathway and antioxidants. We detected a total of 130 negative and 491 positive metabolomic features using a ultrahigh-performance liquid chromatography ion trap time-of-flight mass spectrometry (UHPLC-QTOF-MS). Intramodular significance and real time-quantitative polymerase chain reaction (RT-qPCR) expressions showed that these genes are the true candidate genes. Consortium treated maize had higher jasmonic acid (JA), salicylic acid (SA), and ethylene (ET) levels.ConclusionOur results provide insights into the genetics, biochemicals, and metabolic diversity and are useful for future biocontrol strategies against ACB attacks.

The compatibility between biocontrol agents and pesticides seems to be a sustainable control strategy in agriculture. Therefore, the in vitro compatibility of three native isolates of Trichoderma was evaluated in three concentrations of chlorpyrifos (960, 1200, and 1440 mg/L), by determining the effect on spore germination, mycelial growth, and the antagonistic capacity. The isolates correspond to Trichoderma asperellum TCA 3, Trichoderma asperellum TCA 21 and Trichoderma harzianum TCA 23. Both spore germination and mycelial growth were performed using the poisoned medium method, while the antagonistic capacity was evaluated against Botrytis sp. in a dual culture. The results showed that TCA 21 strain had a higher germination percentage (79.46, 59.79, and 37.43%) than the TCA 3 and TCA 23 strains, in the three concentrations of chlorpyrifos. Regarding the mycelial growth of the three native strains in chlorpyrifos are affected when concentration of chlorpyrifos increase (p < 0.05). Finally, the antagonistic capacity of the three strains was not affected by any concentration of chlorpyrifos, where strains TCA 21 and TCA 23 presented a degree of antagonism of one, while TCA 3 presented a degree of two, according to the scale used by Bell. In conclusion, T. asperellum TCA 21 was the one that presented the best in vitro compatibility with chlorpyrifos at concentrations of 960 and 1200 mg/L, compared to T. asperellum TCA 3 and TCA 23. These results are favorable for field application since these native strains can also have the ability to degrade the insecticide, representing a sustainable and eco-friendly alternative to the environment.

The species of Trichoderma are one of the most frequently used natural biocontrol agent. This study, we identified isolate HbGT6-07 of Trichoderma asperellum and evaluated the antimicrobial effects both in vitro and in silico approaches. Tested 10% concentrated culture filtrate of HbGT6-07 inhibited 98% of colony radial growth in B. cinerea (B05.10) as well as 91% of S. sclerotiorum (A367). HbGT6-07 was detected to produce volatile organic compounds (VOCs) with antifungal activity. In in-vitro dish-within-dish method (DwD), The HbGT6-07 VOCs effectively reduced colonial diameter, growth rate and sclerotia production by two virulent fungal pathogens. Moreover, the hyphal fragments of HbGT6-07 demonstrated successful mycelia growth suppression (97%) against infection oilseed rape leaves by hyphae of the two virulent fungal pathogens through competition. The mixed culture assay, exhibited that the isolate T. asperellum HbGT6-07 was significantly reduced the production and weight of sclerotia. The GC-MS analysis identified 32 VOCs derived from HbGT6-07. In addition, VOCs derived from HbGT6-07 were assessed against targeted protein of three fungal species; Aspergillus oryzae, Saccharomyces cerevisiae, Candida albicans via molecular docking. Butylated hydroxytolune and Beta-Cedrene had energy (−5.3 and −5.7 Kcal/mol) for targeted protein of Aspergillus oryzae and (−6.8 and −8.0 Kcal/mol) for Saccharomyces cerevisiae, whereas alpha-bergamotene and Beta-Cedrene exihbit energy (−7.5 and −7.4 Kcal/mol), respectively. The molecular dynamics study confirms the structural stability and rigidity of the docked complex through multiple descriptors from simulation trajectories. The above findings indicated that HbGT6-07 could attain competitive progress via production of VOCs and comprehensive mycelial growth.

Chitinase là enzyme xúc tác thủy phân chitin bằng cách phân cắt liên kết 1,4-N-acetyl-β-glucosaminide. Chitinase được ứng dụng rộng rãi trong nhiều lĩnh vực khác nhau, đặc biệt trong kiểm soát dịch bệnh, giảm thiểu ô nhiễm, nghiên cứu sinh học cơ bản và ứng dụng. Nguồn thu nhận chitinase chủ yếu là từ vi sinh vật, điển hình từ các chủng nấm Trichoderma. Gen mã hóa chitinase 42 kDa (chi42) của Trichoderma asperellum SH16 sau khi tạo dòng vào vector pUC19 được ghép nối thành công vào vector pQE30 để biểu hiện ở E. coli M15. Chitinase là enzyme ngoại bào. Cấu trúc bậc 2 của chitinase bao gồm 15 chuỗi xoắn α và 13 phiến β với cấu trúc không gian tương đồng cao với enzyme thủy phân chitin ở T. harzianum. Chitinase có khả năng chịu nhiệt độ cao hơn 65 °C và hoạt tính xúc tác mang tính acid. Kết quả nghiên cứu là cơ sở cho các nghiên cứu biểu hiện và sản xuất enzyme tái tổ hợp.

Trichoderma species were known as biological control agents against phytopathogenic fungi because they produce a variety of chitinases. Chitinases are hydrolytic enzymes that break down glycosidic bonds in chitin, a major component of the cell walls of fungi. The present study shows that extracellular chitinase activity reached a maximum value of approximately 22 U/mL after 96 h of T. asperellum PQ34 strain culture. The optimal temperature and pH of enzyme are 40°C and 7, respectively, whereas the thermal and pH stability range from 25°C to 50°C and 4 to 10, respectively. Chitinase at 60 U/mL inhibited nearly completely in vitro growth of Colletotrichum sp. (about 95%) and Sclerotium rolfsii (about 97%). In peanut plants, 20 U/mL of chitinase significantly reduced the incidence of S. rolfsii infection compared to controls. The fungal infection incidence of seeds before germination and 30 days after germination was only 2.22% and 2.38%, while the control was 13.33% and 17.95%. Besides, chitinase from T. asperellum PQ34 can also prevent anthracnose that is caused by Colletotrichum sp. on both mango and chilli fruits up to 72 h after enzyme pre-treatment at 40 U/mL. In mango and chilli fruits infected with anthracnose, 40 U/mL dose of chitinase inhibited the growth of fungi after 96 h of treatment, the diameter of lesion was only 0.88 cm for mango and 1.45 cm for chilli, while the control was 1.67 cm and 2.85 cm, respectively.

El objetivo de esta investigación fue evaluar el crecimiento de Trichoderma asperellum en medio sólido utilizando como única fuente de carbono a los plaguicidas cipermetrina (piretroide) y clorpirifos (organofosforado). Para este propósito, se realizaron cultivos de T. asperellum sobre el medio Agar Mínimo de Sales, los cuales tenían como únicas fuentes de carbono a los plaguicidas cipermetrina (480 ppm) y clorpirifos (250 ppm). Posteriormente los cultivos fueron incubados a 25 ºC por 5 días. Finalmente se calcularon las velocidades de crecimiento. El control tuvo como única fuente de carbono a la glucosa. Los resultados muestran que T. asperellum puede crecer en ambos medios utilizando a los plaguicidas como una sola fuente de carbono y en cuanto a sus velocidades de crecimiento fueron 2.88 ± 0.05 mm/día (cipermetrina) y 2.74 ± 0.05 mm/día (clorpirifos). En conclusión, T. asperellum es capaz de utilizar a los plaguicidas cipermetrina y clorpirifos mediante procesos catabólicos y tiene el potencial de ser usado en biorremediación de plaguicidas de suelos contaminados.

Of diseases affecting maize (Zea mays), Fusarium graminearum is one of the most common pathogenic fungi that cause stalk rot. In the present study, the Trichoderma asperellum GDFS1009 strain was shown to be an effective biocontrol agent against stalk rot. In a confrontation culture test, Trichoderma strain displayed an approximately 60% inhibition rate on the mycelial growth of F. graminearum. In pot trials, the application of 2 g/pot of T. asperellum GDFS1009 granules had the best control effect on stalk rot at the seedling stage (up to 53.7%), while the average plant height and fresh weight were also significantly improved. Additionally when fertilizer was added at 8 g/pot, the application of 3 g/pot of Trichoderma granules had the best control effect on maize stalk rot (40.95%). In field trials, when inoculating F. graminearum alone, the disease index for inoculating was 62.45, but only 31.43 after treatment with T. asperellum GDFS1009 granules, suggesting a control efficiency of 49.67%. Furthermore, in a naturally F. graminearum-infected field, Trichoderma granules, when applied for 3 consecutive years, showed significant control of stalk rot and increased yields.

Velvet family proteins are global regulators of fungal growth and development. Here, we reported the role of Vel1 and Lae1 from T. asperellum in osmotic tolerance. Deletion of the Vel1 and Lae1 genes led to the retardation of vegetative mycelial growth under saline conditions. The strain carrying the overexpression locus of the Vel1 and Lae1 genes was highly resistant to oxidative stress by upregulating the enzymes and genes involved in antioxidant activity. Major physiological changes in the cell wall and vacuoles occurred under high saline conditions. The Vel1 and Lae1 overexpression strains increased cell wall thickness and the number of vacuoles, which seems to lead to an increase of the osmolyte content of glycerol and proline. The absorption of Na+ content in the vacuole of the Vel1 and Lae1 overexpression strains was increased, while the absorption of Na+ was impaired in the Vel1 and Lae1 knock out strains, in which the Na+ was localized in the cell wall membrane. This result supported the significant correlation of the expression of genes with the ionic transportation in T. asperellum. Maize root colonization by the Vel1 and Lae1 gene overexpression strain was increased, which would mitigate the stress caused by the absorption of Na+ in the maize roots and increased the plant growth. Our results highlighted the importance of Vel1 and Lae1 proteins to the salinity stress tolerance of T. asperellum and the mitigation of Na+ stress to plants for sustainable agriculture.

Enzyme phytase được sử dụng phổ biến trong chăn nuôi nhằm giúp vật nuôi hấp thu tốt phosphor và kiểm soát các yếu tố kháng dưỡng. Trong số 4 chủng vi nấm khảo sát, Trichoderma asperellum cho thấy việc thu nhận phytase hiệu quả nhất trên môi trường bán rắn chứa 60% bột bắp và 40% bã đậu nành. Ảnh hưởng của tỷ lệ dịch khoáng bổ sung và thời gian nuôi được tối ưu theo phương pháp đáp ứng bề mặt (Response surface methodology, RSM). Ở tỷ lệ dịch khoáng bổ sung 60,4%, thời gian nuôi cấy 3,75 ngày, chế phẩm có hoạt độ phytase đạt 5,31±0,65 UI/g. Chế phẩm thủy phân phytate trong thức ăn viên tốt nhất ở pH 5,0 với hàm lượng phosphor vô cơ tăng 25,6% sau 2 giờ. Thử nghiệm bổ sung 1% (w/w) chế phẩm giàu phytase vào khẩu phần ăn gà thả vườn cho thấy hàm lượng phosphor vô cơ trong phân giảm từ 14,5% đến 39,1% so với đối chứng, trong khi khối lượng gà không khác biệt, đạt 791,5 g/con sau 7 tuần.

Trichoderma spp. is a fungus widely used to control soil-borne pathogens, such as Rhizoctonia solani which is plant pathogenic fungi in widely host range, especially on rice. This research aimed to evaluate the ability of Trichoderma asperellum isolate UGM-LHAF against R. solani causing sheath blight disease of rice in vitro condition. Trichoderma sp. used in this research was obtained from The Biological Laboratory of Pakem, Yogyakarta, Indonesia, and Rhizoctonia sp. was obtained through isolation of diseased rice obtained from rice fields in Yogyakarta. The two isolates were characterized base on morphology and molecular identification based on ITS rDNA. The pathogenicity test of Rhizoctonia sp. was evaluated by adding four sclerotia of Rhizoctonia sp. near rice roots at 6 days after sowing. The in vitro test used dual culture and antifungal activity (0%, 10%, 25%, 50% culture filtrate of Trichoderma sp.) with three replicates of each treatment. Two isolates were identified as T. asperellum and R. solani. Sheath blight symptoms appeared after 12 days inoculation. In the in vitro test, T. asperellum isolate UGM-LHAF was able to inhibit the mycelial growth of R. solani (64.23% on dual culture and 68.5% on antifungal activity). This study suggests that T. asperellum isolate UGM-LHAF able to inhibit the growth of R. solani and can be a further potential candidate as a biocontrol agent against R. solani causing sheath blight disease of rice.