Trichoderma Isolates Research Articles

EFFECTIVENESS OF TRICHODERMA SPECIES IN THE MANAGEMENT OF SUGARCANE WILT (FUSARIUM SACCHARI)

Sugarcane is a major cash crop in Pakistan. It has a great contribution in world’s economy. Many diseases cause many losses in yield of sugarcane. Fungi are considered as abundant traitorous pathogens that obstruct the viable growth of sugarcane crop. Wilt disease caused by fungal pathogen Fusarium sacchari caused many losses in sugarcane production in Pakistan. Fusarium sacchari is a causative fungus of wilt in sugarcane. Biocontrol agent Trichoderma spp. used in the management of sugarcane wilt. These bio‐agents help in diseases management and play important role in increasing the crop yield. For this reason, these bio‐agents are used for biological maintaenace of crop diseases and research globally. The use of PDA medium in vitro revealed antagonistic activity of various Trichoderma isolates against the sugarcane wilt pathogen Pathogenicity test also executed to check the most aggressiveness strain of Fusarium spp. to cause disease in sugarcane plant. In vivo, Trichoderma multiply on different substrates and corn Cobbs is good byproduct Trichoderma multiply on it and mix in soil of pots. Pot experiment performed with different treatments to control wilt disease by using of antagonist’s to check disease incidence and disease reduction.

Management of Leaf Blight Disease of Medicinal Plants with Bio-agents and Chemical Fungicides

Leaf blight affecting medicinal plants like Piper longum, Tylophora indica, and Hibiscus subdariffa is a major issue in West Bengal, causing significant damage year-round. Effective management of this disease is crucial. Several chemical fungicides and Trichoderma isolates were evaluated against the disease of these plants under field condition. Carbendazim and mancozeb were found to be effective in Piper longum in terms of higher disease control (47.50 & 41.47%) with lower per cent disease incidence (49.87 & 51.47%) and per cent disease index (25.34 & 28.25%) over the control. Although all the Trichoderma isolates were not effective as chemical fungicides but Trichoderma isolate-3 was found to have more efficiency in disease control (35.23%) compared to other two isolates. Mancozeb was treated as the most effective fungicide in T. indica to control the disease (45.07%) with lower disease incidence (54.87%) and disease severity (27.34%) compared to other treatment and control plot. In contrary in H. subdariffa carbendazim was found the most effective fungicide with lowest disease incidence (47.89%) and disease severity (23.37%) resulting higher disease control (47.81%) compared to control. It was followed by cupper oxychloride and mancozeb comparatively with higher disease incidence (49.79 & 51.43%) and disease severity (25.89 & 31.78%). Whether all the Trichoderma isolates did not show significant efficacy as fungicides but Trichoderma isolate-3 showed comparatively good result in disease control (34.32%) over other two isolates and control plot. So, the aforementioned respective fungicides could be used in the control of leaf blight disease of respective medicinal plant. But to control the disease with bio agents more efficient Trichoderma culture need to be investigated.

Indigenous Trichoderma isolates from Bukit Kijang, North Sumatra, Indonesia, are biocontrol agents against Ganoderma boninense in infected oil palm seedlings

Basal stem rot, caused by Ganoderma boninense is a serious disease in oil palm plantations in North Sumatra, Indonesia. However, the use of fungicides to control this pathogen can cause instability in terrestrial ecosystems and decrease the number of beneficial microorganisms. Therefore, identifying other effective and environmentally friendly control methods, such as Trichoderma, is essential. Trichoderma was previously isolated from the leaf, rachis, stem, and root tissues of oil palms but not from the rhizosphere soil. Therefore, this study aimed to assess the potency of indigenous Trichoderma isolates from the rhizosphere soil of oil palm seedling plantations in Bukit Kijang, North Sumatra, Indonesia, as biocontrol agents to reduce G. boninense infection. Four Trichoderma isolates were isolated from healthy oil palm rhizospheres in Bukit Kijang and screened in vitro. Based on molecular identification, the Trichoderma isolates were identified as T. asperellum1, T. virens1, T. asperellum2, and T. virens2. All Trichoderma species inhibited the development of Ganoderma and stimulated the vegetative growth of oil palm seedlings. Therefore, T. asperellum and T. virens can be used as biocontrol agents to control G. boninense and stimulate the vegetative growth of oil palm seedlings. To our knowledge, this is the first report of indigenous Trichoderma isolates from Bukit Kijang that can be used as biocontrol agents to control G. boninense.

Genome sequencing and functional analysis of potential Trichoderma species for controlling Pythium schmitthenneri-Induced root rot in olive trees

Root rot disease caused by Pythium schmitthenneri is a significant challenge in olive (Olea europaea L.) cultivation. Due to the limitations of chemical control, this study explores the potential of Trichoderma species as a sustainable alternative with an overview in the genetic properties involved in it. Eight Trichoderma isolates were obtained from olive roots and identified using rDNA internal transcribed spacer (ITS) sequences, revealing four species: T. harzianum, T. longibrachiatum, T. virens, and T. viride. Among these, T. harzianum T-BM6 and T. virens T-KH4, showed 81.4 and 78.77 % inhibition rates of pathogen growth, respectively, in vitro. Greenhouse trials demonstrated that these isolates reduced disease severity to 18.75 % for T-BM6 and 31.25 % for T-KH4. All isolates produced cellulase, with T-BM6 exhibiting high cellulolytic activity. Genomic analysis highlighted key genes related to the MAPK signaling pathway, sulfur metabolism, and inositol phosphate metabolism in T-BM6, and phenylalanine, tyrosine, and tryptophan metabolism in T-KH4. These results suggest that T. harzianum T-BM6 and T. virens T-KH4 are promising candidates for biological control, offering a sustainable alternative to chemical treatments for managing olive root rot.

Characterization of Trichoderma spp. and their antagonistic activity against soilborne fungi associated with chickpea wilt in Sinaloa, Mexico

Chickpea wilt, caused by a complex of soilborne fungi (Fusarium spp. Macrophomina phaseolina, Rhizoctonia solani, Sclerotium rolfsii, and Sclerotinia sclerotiorum), is the most important disease of chickpeas (Cicer arietinum) in Mexico. The aims of this study were to characterize Trichoderma isolates using a combination of phenotypic and molecular approaches and to evaluate their antagonistic activity against soilborne fungi associated with chickpea wilt. A total of 30 Trichoderma isolates were obtained from rhizospheric soil samples collected from chickpea fields in different locations of Sinaloa, Mexico. Dual confrontation assays showed the potential antagonistic effect of Trichoderma isolates against F. languescens, M. phaseolina, R. solani, S. rolfsii, and S. sclerotiorum. Five Trichoderma isolates (FAVF335, FAVF340, FAVF345, FAVF349, and FAVF351) exhibited mycelial growth inhibition of the five pathogens that ranged from 56 to 71%. These isolates were characterized using cultural, morphological, and molecular studies and tested in vivo for their ability to control soilborne pathogens in two chickpea cultivars (Blanco Sinaloa-92 and P2245) under greenhouse conditions. Phylogenetic analysis based on a combined ITS, EF-1α, and rpb2 sequence dataset identified T. afroharzianum (FAVF345, FAVF349, and FAVF351) and T. longibrachiatum (FAVF335 and FAVF340). Coating of chickpea seeds with T. longibrachiatum (FAVF335 and FAVF340) significantly reduced the disease severity and improved the plant growth-promoting attributes.

Assessment of Trichoderma Species Isolated From Volcanic Soil of a Durian Field in Sisaket Province, Thailand for Plant Growth Promotion and Biocontrol Potential

Trichoderma species are ubiquitous saprophytic fungi commonly found in soil, with potential as fungal biocontrol agents for plant disease control and growth promotion. This research aimed to assess the attributes of Trichoderma sp. as plant growth promoters, focusing on nitrogen fixation, siderophore production, plant nutrient solubilization and indole-3-acetic acid production. Out of 19 isolates tested, 6 (T05, T19, T25, T27, T28 and T33) showed superior plant growth promotion abilities. These isolates were further evaluated for their capacity to enhance the germination of Khao Dawk Mali 105 rice seeds. Seeds were soaked in a suspension of conidia at a concentration of 1×1012 conidia/mL, and germination rates were measured. The germination rate ranged between 77.67 - 86.67 %, statistically significantly higher than the control rate of 33.33 %. Among the isolates, T05 exhibited the highest promotion of plant growth, evidenced by root length (7.27 cm), shoot length (8.50 cm), fresh weight (0.54 g) and dry weight (0.27 g), all significantly different from the control. Additionally, the potential of isolates to inhibit the fungal pathogen Colletotrichum sp. MN-3 was assessed using a dual culture method, with T35 showing the highest inhibition percentage (60.90 %). Morphological classification and nucleotide sequencing of the ITS1-5.8S-ITS2 region of rDNA gene identified the isolates as T. harzianum, T. reesei, T. asperellum and T. longibrachiatum. These findings underscore the effectiveness of Trichoderma sp. in promoting plant growth and suppressing plant pathogenic fungi. HIGHLIGHTSSix Trichoderma isolates (T05, T19, T25, T27, T28 and T33) exhibited superior plant growth promotion abilities. These isolates significantly enhanced germination rates of Khao Dawk Mali 105 rice seeds compared to the method. Among the isolates, T05 showed the greatest promotion of plant growth, as evidenced by root length, shoot length, fresh weight and dry weight. Trichoderma isolates demonstrated potential for inhibiting the fungal pathogen Colletotrichum sp. MN-3. Morphological classification divided the Trichoderma sp. into 4 groups, and species identification through nucleotide sequencing revealed T. harzianum, T. reesei, T. asperellum and T. longibrachiatum. GRAPHICAL ABSTRACT

New Trichoderma Strains Suppress Blue Mold in Oranges by Damaging the Cell Membrane of Penicillium italicum and Enhancing Both Enzymatic and Non-Enzymatic Defense Mechanisms in Orange Fruits

Blue mold disease, caused by Penicillium italicum (P. italicum), presents a significant challenge to orange fruits (Citrus sinensis L.) and other citrus crops globally. Biological control, particularly Trichoderma species, offers a promising alternative to synthetic fungicides. Therefore, this study aimed to isolate, identify, and evaluate the antagonistic activities of two Trichoderma isolates against P. italicum. These isolates were molecularly identified and assigned accession numbers PP002254 and PP002272, respectively. Both isolates demonstrated significant antifungal activity in dual culture assays. Moreover, the culture filtrates (CFs) of Trichoderma longibrachiatum PP002254 and Trichoderma harzianum PP002272 suppressed the mycelial growth of P. italicum by 77.22% and 71.66%, respectively. Additionally, CFs reduced the severity of blue mold on orange fruits by 26.85% and 53.81%, compared to 100% in the control group. Scanning electron microscopy revealed that treated P. italicum hyphae were shrunken and disfigured. Enzyme activities (catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase) in treated oranges increased, along with total soluble phenolics and flavonoids. Conversely, malondialdehyde (MDA) levels decreased in treated fruits. These findings suggest that T. longibrachiatum PP002254 and T. harzianum PP002272 could be effective biocontrol agents for managing blue mold and other citrus postharvest diseases.

Molecular characterization of plant growth-promoting Trichoderma from Saudi Arabia.

Fungi in the genus Trichoderma are widespread in the environment, mainly in soils. They are used in agriculture because of their mycoparasitic potential; Trichoderma have the ability to increase plant health and provide protection against phytopathogens, making them desirable plant symbionts. We isolated, identified, and characterized Trichoderma from different regions of Saudi Arabia and evaluated the ability of Trichoderma to promote plant growth. Morphological and molecular characterization, along with phylogenetic studies, were utilized to differentiate between Trichoderma species isolated from soil samples in the Abha and Riyadh regions, Saudi Arabia. Then, plant growth-promoting traits of the isolated Trichoderma species were assessed. Eight Trichoderma isolates were characterized via morphological and molecular analysis; six (Trichoderma koningiopsis, Trichoderma lixii, Trichoderma koningii, Trichoderma harzianum, Trichoderma brevicompactum, and Trichoderma velutinum) were from Abha and two (T. lixii and T. harzianum) were from Riyadh. The isolated Trichoderma strains belonged to three different clades (Clade 1: Harzianum, Clade 2: Brevicompactum, and Clade 3: Viride). The Trichoderma isolates varied in plant growth-promoting traits. Seeds treated with most isolates exhibited a high percentage of germination, except seeds treated with the T3-T. koningii isolate. 100% germination was reported for seeds treated with the T4-T. harzianum and T6-T. brevicompactum isolates, while seeds treated with the T1-T. koniniopsis and T5-T. lixii isolates showed 91.1% and 90.9% germination, respectively. Seeds treated with the T8-T. velutinum, T2-T. lixii, and T7-T. harzianum isolates had germination rates of 84.1%, 82.2%, and 72.7%, respectively. The Trichoderma isolate T5-T. lixii stimulated tomato plant growth the most, followed by T7-T. harzianum, T8-T. velutinum, T4-T. harzianum, T1-T. koniniopsis, T2-T. lixii, and T6-T. brevicompactum; the least effective was T3-T. koningii. A maximum fresh weight of 669.33mg was observed for the T5-T. lixii-treated plants. The Abha region had a higher diversity of Trichoderma species than the Riyadh region, and most isolated Trichoderma spp. promoted tomato growth.

Characterization and quantification of peptaibol produced by novel Trichoderma spp: Harnessing their potential to mitigate moisture stress through enhanced biochemical and physiological responses in black pepper (Piper nigrum L.).

Trichoderma spp. is primarily applied to manage biotic stresses in plants. Still, they also can mitigate abiotic stresses by the stimulation of antioxidative protective mechanisms and enhanced synthesis of secondary metabolites. The study optimized the conditions to enhance peptaibol production by novel Trichoderma spp, characterized and quantified peptaibol- alamethicin using HPLC and LC MS-MS. The present study investigated these isolates efficacy in enhancing growth and the associated physio-biochemical changes in black pepper plants under moisture stress. Under in vitro conditions, out of 51 isolates studied, six isolatesviz., T. asperellum (IISR NAIMCC 0049), T. erinaceum (IISR APT1), T. harzianum (IISR APT2), T. harzianum (IISR KL3), T. lixii (IISR KA15) and T. asperellum (IISR TN3) showed tolerance to low moisture levels (5, 10 and 20%) and higher temperatures (35 and 40°C). In vivo evaluation on black pepper plants maintained under four different moisture levels (Field capacity [FC]; 75%, 50%, and 25%) showed that the plants inoculated with Trichoderma accumulated greater quantities of secondary metabolites viz., proline, phenols, MDA and soluble proteins at low moisture levels (50% and 25% FC). In the present study, plants inoculated with T. asperellum and T.harzianum showed significantly increased growth compared to uninoculated plants. The shortlisted Trichoderma isolates exhibited differences in peptaibol production and indicated that the peptide might be the key factor for their efficiency as biocontrol agents. The present study also demonstrated that Trichoderma isolates T. harzianum and T. asperellum (IISR APT2 & NAIMCC 0049) enhanced the drought-tolerant capabilities of black pepper by improving plant growth and secondary metabolite production.

Effect of Trichoderma Culture Filtrates of as Inducer on Growth Parameters of Tomato (Solanum lycopersicum L.) against Fusarium Wilt

Aims: The present study was carried out to evaluate the effect of culture filtrates from different Trichoderma isolates at 10% concentration on the growth parameters of tomato plants over two growing seasons (2021-22 and 2022-23). Study Design: The experiment was performed both in vitro and in vivo condition, Completely Randomized Design (CRD) used for in vitro and Randomized Block Design (RBD) used for in vivo experiment Place and Duration of Study: The experiment was performed at Chandra Shekhar Azad University of Agriculture and Technology, Kanpur (India) during 2021-22 and 2022-23. Methodology: The soil sample collected from 12 different location of India and Trichoderma isolated and these different isolates used to produce their individual culture filtrate and these culture filtrates further use with 10% concentration as treatment for tomato plant. The effect of culture filtrates on plant height, root length, number of branches, fresh and dry shoot and root weight were observed under the experiment. Results: Results show significant improvements in plant height, root length, fresh and dry weights and number of branches in all culture filtrate treated plants involving T9 (seedling treatment + one foliar spray with culture filtrate of Trichoderma Nagaur), which consistently outperformed other isolates. Plant height increased to 43.06 cm in 2021-22 and 44.56 cm in 2022-23 at 40 days after transplanting with isolate T9 treatment followed by culture filtrate of Trichoderma Jabalpur (T7) and Trichoderma Banda (T6) showed 11.56, 22.96, 33.72, and 37.15 cm in 2021-22, and 11.38, 23.24, 34.46 and 36.08 cm for T6 in the same year. Among all the culture filtrates, the treatment T9 significantly enhances root length, fresh shoot and root weights and the number of branches. The plants treated with isolate T9 also exhibited an increase of 98.94% in root fresh weight and 42.31% in shoot weight. Conclusion: The findings confirm the growth-promoting potential of the T9 Trichoderma, attributed to mechanisms such as hormone production, enhanced nutrient uptake and improved root architecture. These results align with previous studies demonstrating the benefits of Trichoderma species in plant growth enhancement.

Morphological and Molecular Characterization of Trichoderma Isolates from Vegetable Crop Rhizospheres in Nepal

Background Trichoderma spp. hold significant potential as biocontrol agents in agriculture due to their antagonistic properties against plant pathogens. The study aimed to characterize and identify Trichoderma isolates from rhizospheric regions of vegetable crops. Methods In this study, Trichoderma isolates were collected from rhizospheric soil samples of vegetable crops from different ecological zones and were selected for comprehensive morphological and molecular characterization. The isolates were visually assessed for colony color, growth pattern, aerial mycelium presence, phialide and conidial morphology, and chlamydospore presence. Molecular analysis was employed based on ITS and tef-1α sequences. Diversity indices were also computed for different ecological zones. Results The morphological characteristics and phylogenetic trees for both regions provided a clear species resolution, with four main clades: Harzianum, Viride, Brevicompactum and Longibrachiatum with 12 species T. harzinaum, T. afroharzianum, T. lentiforme, T. inhamatum, T. camerunense, T. azevedoi, T. atroviride, T. asperellum, T. asperelloides, T. koningii, T. longibrachiatum and T. brevicompactum and nine species as a new country record. Diversity indices indicated that high mountain regions displayed the highest species diversity and evenness (H = 1.724 [0.28], J = 0.84, D = 0.28), followed by hilly regions (H = 1.563 [0.28], J = 0.72, D = 0.28). Plains, on the other hand, exhibited lower species diversity (H = 1.515, J = 0.66, D = 0.33). The calculated species abundance values showed that plains (E = 2.11), mid-hills (E = 1.95), and high mountains (E = 1.99) each had their unique diversity profiles. Notably, T. afroharzianum and T. asperellum were predominant. Conclusions Overall, the study unveiled a rich diversity of Trichoderma species in different agricultural zones of Nepal. These findings shed light on the ecological distribution and diversity of Trichoderma spp., which could have significant implications for sustainable agriculture and biological control strategies.

Effect of Cow Urine and Culture Filtrates of Trichoderma isolate S-13 on Fungal Pathogens of Basmati Rice

Rice (Oryza sativa L.) is the principle food crop for majority population of the world. Basmati rice is a unique crop of Himalaya comprising good quality characters and aroma. Basmati rice is being infected by numerous diseases. To manage these diseases, fungicides being used for years. This leads to the residual effects of fungicides on grains and soils so there is needs for alternate approaches to the manage disease. Hence use of cow urine and Trichoderma isolate S-13 is considered as the sustainable approach for management of important diseases of basmati rice. In this study we used different concentration of cow urine, Trichoderma isolate S-13, and Propiconazole (control check) against brown spot and bakanae disease of rice. Among these the maximum mycelial inhibition observed in Trichoderma isolate S-13 @15per cent (66.33per cent) followed by cow urine @15per cent (52.59 per cent). In Bakanae disease the maximum inhibition observed in Trichoderma isolate S-13 @ 15per cent (58.89 per cent) followed by cow urine@15per cent (49.26 per cent).

Harnessing Trichoderma spp.: A Promising Approach to Control Apple Scab Disease.

Apple scab, caused by the pathogenic fungus Venturia inaequalis, can result in significant economic losses. The frequent use of fungicidal products has led to the emergence of isolates resistant to commonly used active substances. Therefore, biological control offers a sustainable alternative for managing apple scab. In this study, eight Trichoderma isolates were evaluated against five different isolates of V. inaequalis isolated from the Fes-Meknes region. The biocontrol potential of these Trichoderma isolates had previously been demonstrated against other pathogens. The results indicated that the inhibition rate of mycelial growth of V. inaequalis obtained with Trichoderma spp. isolates ranged from 50% to 81%, with significant differences observed among the pathogenic isolates after 5 and 12 days of incubation. In addition, the in vitro tests with Trichoderma cell-free filtrates showed inhibition rates ranging from 2% to 79%, while inhibition rates ranged from 5% to 78% for volatile compound tests. Interestingly, the inhibition of spore germination and elongation was approximately 40-50%, suggesting the involvement of antifungal metabolites in their biocontrol activities. The in vivo bioassay on detached apple leaves confirmed the biocontrol potential of these Trichoderma isolates and demonstrated their ability to preventively control apple scab disease. However, their efficacies were still lower than those of the fungicidal product difenoconazole. These findings could contribute to the development of an effective biofungicide based on these Trichoderma isolates for reliable and efficient apple scab control.

Trichoderma harzianum TIND02 upregulates the expression of pathogenesis-related genes and enzymes and enhances gray blight resistance in tea

The gray blight incited by Pestalotiopsis and allied genera is a prevalent disease affecting tea cultivation, and managing it with Trichoderma spp. is an alternative to synthetic fungicides. Plants modify their arsenal system against pathogens when they are exposed to Trichoderma spp., which produces proteins and enzymes associated with pathogenesis. Understanding the expression pattern of defense-related markers will help in developing gray blight resistance tea cultivars. Thus, this study intended to induce resistance against gray blight in tea by Trichoderma harzianum TIND02. For this, a total of eight Trichoderma isolates originated from organic tea rhizospheres were characterized and evaluated for their efficacy. Dual culture test revealed isolate TIND02 as the most potential candidate with 74.6% inhibitory activity against gray blight pathogen Pseudopestalotiopsis theae. Molecular characterization based on ITS and tef-1 alpha genes confirmed isolate TIND02 as T. harzianum. Scanning electron microscopic study showed the mycoparasitic nature of T. harzianum TIND02 (TH-TIND02) to Ps. theae. The ethyl acetate extract of TH-TIND02 at 100 and 200 μg mL−1 showed potential inhibitory activity (>69.9%) against Ps. theae which confirmed the presence of higher volatile metabolites. Gas chromatography–Mass spectrometry study revealed that ethyl acetate extract of TH-TIND02 was composed of 21 major and minor volatile organic compounds with acetamide, 2, 2, 2-trifluoro-N, N-bis trimethyIsilyl–C (94.74%) as a major component. The isolate also produced chitinase, cellulase, β-1, 3 glucanase, and protease hydrolytic enzymes. Nursery experiments revealed that 2% and 5% doses (2 × 106 CFU mL−1) of TH-TIND02 significantly reduced respective 65.0% and 70.0% disease severity over control with improved plant growth. Besides, expressions of defense-related enzymes (chitinase, pHenolics, peroxidase, phenylalanine ammonia lyase, β-1, 3-glucanase, and polyphenol oxidase) and pathogenesis-related genes (chitinase and β-1, 3-glucanase) due to TH-TIND02 were determined. The secretion of defense-related enzymes was highly upregulated in plants applied with TH-TIND02 followed by Ps. theae inoculation compared to controls. The RT-qPCR analysis showed that the expression of both genes in co-inoculated plants was two-fold higher than in control after 21-day post incubation. These results suggest that TH-TIND02 application reduced gray blight severity by elevated enzyme activity and overexpressed pathogenesis-related genes in tea plants which offer for its eco-friendly and sustainable use as a bio-fungicide in tea gardens.

Trichoderma Isolates Against Abiotic Stresses and Management of Collar rot of Lentil (Lens culinaris L.) Caused by Sclerotium rolfsii.

A total of 30 native Trichoderma isolates were collected from the Agricultural Research Station, Ummedganj- Kota, Rajasthan, India. Out of which 9 native isolates were evaluated for bio-efficacy against Sclerotium rolfsii. Isolate ARS K-21 exhibited maximum inhibition (89.26%), followed by ARS K-11 (83.70%) in dual culture. Subsequent evaluations revealed the compatibility of efficient isolate ARS K-21 with various bio botanicals displaying minimum inhibition with Vermiwash (1.11-3.70%) followed by Beejamarat (0.38-15.92%) and Brahmastra (7.78-19.68%), while ARS K-11 displayed compatibility onlywith Dasparni ark with a minimum inhibition of 1.11-3.70%. Assessment of abiotic stress tolerance of the isolates revealed that most isolates thrived at 200 mM and 400 mM NaCl salt concentrations, with ARS K-21 and ARS K-24 demonstrating moderate growth levels across higher concentrations, except at 1200 mM. Optimal growth of the isolates occurred at 25and 30°C, with deviations leading to growth inhibition. Isolates ARS K-1, ARS K-11, ARS K-12 and ARS K-21 exhibited resilience to temperature extremes. ARS K-21 has shown exceptional growth proficiency across a wide pH spectrum (pH 5 to 8.5) followed by ARS K- 24, highlighting their versatility. Mass multiplication of efficient isolate ARS K-21 enriched with vermicompost led to the standardization of a dosage (30g/kg soil) for managing collar rot in lentil crops at 5g inoculum per kg soil of S. rolfsii.

Biological suppression of sugarcane smut through native Trichoderma isolates increased sugarcane yield at Wonji, Ethiopia

Smut disease is identified as the main constraint in the cultivation of sugarcane in Ethiopia. Although the country has employed a range of management strategies, none of them have resulted in ecologically safe and sustainable control. The use of Trichoderma spp. in the management of this disease is one of the unexploited potentials under Ethiopian conditions. Native Trichoderma isolates – MI3-m33 (Trichoderma viride), MKI2-k17 (T. asperellum), MKI3 (T. harzianum), WI12-25 (T. viride), and WI4-27 (T. harzianum) – were verified for their effectiveness in managing smut pathogen in vitro at Wonji. However, the isolates’ effect on smut under in vivo conditions was unknown. Therefore, this experiment was conducted to evaluate these isolates against smut. The impact of seed cane treatment with the isolates against smut was evaluated in a randomised complete block design with three replications at Wonji for two cropping seasons. Data on smut intensity and sugarcane yield components were collected. Cane and sugar yield increased by 55.52, 38.33 and 66.78%, and 59.42, 38.69 and 70.42%, respectively due to seed cane treatment by: MKI2-k17, MKI3 and WI4-27. These isolates also guaranteed complete protection against smut. Therefore, these isolates could be used as an alternative to fungicide, following field efficacy and compatibility tests.

The effects of EDTA and Trichoderma species on growth and Cu uptake of maize (Zea mays) plants grown in a Cu-contaminated soil.

Metal contamination in soil poses a significant environmental concern worldwide, necessitating effective remediation strategies such as phytoremediation. The present study investigated the effects of EDTA dosage (1.5 and 3mmolkg-1) and two Trichoderma species (T. harzianum and T. aureoviride) on copper (Cu) content and growth of maize plants grown in a Cu-contaminated soil, as well as Cu fractionation in the soil. In the absence of EDTA, only inoculation with T. harzianum led to a significant increase in shoot biomass. Combining fungal inoculum with EDTA only yielded a significant increase in shoot biomass when using T. aureoviride at a low EDTA rate, highlighting the interplay between fungal species and EDTA rates on plant growth. Results also indicated that EDTA application increased Cu bioavailability, enhancing Cu dissolution and root (not shoot) Cu concentrations. Conversely, inoculation with both Trichoderma species reduced Cu mobility and bioavailability in soil, thereby decreasing the shoot Cu concentrations of plants. When combined with EDTA, only application of T. harzianum resulted in an enhanced shoot Cu concentration, whereas combined application of T. aureoviride and EDTA did not make a significant change compared to the corresponding control (no fungal inoculation, no EDTA), possibly due to a lower compatibility of the T. aureoviride isolate with EDTA. Our results demonstrated that EDTA application, in both non-inoculated and inoculated treatments, increased Cu availability by facilitating its redistribution and transformation from less plant-available fractions (residual, Fe/Mn oxide-bound, and carbonate-bound) to the more readily plant-available forms (water-soluble and exchangeable fractions). In conclusion, although individual Trichoderma application proved beneficial for phytostabilization by reducing Cu content and mitigating Cu toxicity in plants, the combined application of EDTA and a compatible Trichoderma isolate (here, the T. harzianum isolate) holds promise for enhancing the phytoextraction capacity of plants. Although using maize has the advantage of being a food crop, to optimize phytoextraction, plant species with superior metal tolerance and phytoextraction capabilities should be selected, exceeding those of maize.

Potential biological control agents of Geosmithia morbida restrict fungal pathogen growth via mycoparasitism and antibiosis

Thousand cankers disease of Juglans (walnut) and Pterocarya (wingnut) spp. (Fagales: Juglandaceae) is caused by the fungal pathogen Geosmithia morbida Kolarík, Freeland, Utley, and Tisserat (Hypocreales: Bionectriaceae) and bark beetle pest/vector, Pityophthorus juglandis Blackman (Coleoptera: Curculionidae). To further the development of biological management strategies for thousand cankers disease, we assessed the ability of 14 endophytic Trichoderma (Hypocreales: Hypocreaceae) isolates and the commercially available isolate T. afroharzianum strain KRL-AG2 to inhibit the in vitro growth of three different G. morbida isolates via mycoparasitism and antibiosis. To identify factors that may affect field success of candidate biological control agents, we quantified the growth responses of Trichoderma spp. and the commercially available entomopathogenic fungus, Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Cordycipitaceae) strain GHA, to the plant secondary metabolite and antimicrobial compound, juglone in vitro. A total of 12 Trichoderma isolates (from six different Trichoderma species) demonstrated antagonistic activity towards G. morbida in dual-plate assays. Juglone consistently reduced the growth of B. bassiana strain GHA and 14 of the 15 screened Trichoderma isolates in vitro. Additionally, one metabolite-producing Trichoderma isolate, TN4-47, completely inhibited the growth of all three G. morbida isolates across all tested metabolite concentrations and had comparatively greater tolerance to juglone compared to other Trichoderma isolates. Future lines of research should focus on characterizing the active antagonistic compound present in the metabolite filtrates, determine the mode of action of the active component(s), and elucidate how abiotic and biotic factors may influence the growth, persistence, and antagonistic activity of candidate biological control agents in planta.

Exploring the Antifungal Activity of Moroccan Bacterial and Fungal Isolates and a Strobilurin Fungicide in the Control of Cladosporium fulvum, the Causal Agent of Tomato Leaf Mold Disease.

The causal agent of tomato leaf mold, Cladosporium fulvum, is prevalent in greenhouses worldwide, especially under high humidity conditions. Despite its economic impact, studies on antifungal agents targeting C. fulvum remain limited. This study evaluates biocontrol agents (BCAs) as alternatives to chemical controls for managing this disease, alongside the strobilurin fungicide azoxystrobin. From a Moroccan collection of potential BCAs, five bacterial isolates (Alcaligenes faecalis ACBC1, Pantoea agglomerans ACBC2, ACBP1, ACBP2, and Bacillus amyloliquefaciens SF14) and three fungal isolates (Trichoderma spp. OT1, AT2, and BT3) were selected and tested. The in vitro results demonstrated that P. agglomerans isolates reduced mycelial growth by over 60% at 12 days post-inoculation (dpi), while Trichoderma isolates achieved 100% inhibition in just 5 dpi. All bacterial isolates produced volatile organic compounds (VOCs) with mycelial inhibition rates ranging from 38.8% to 57.4%. Likewise, bacterial cell-free filtrates significantly inhibited the pathogen's mycelial growth. Greenhouse tests validated these findings, showing that all the tested isolates were effective in reducing disease incidence and severity. Azoxystrobin effectively impeded C. fulvum growth, particularly in protective treatments. Fourier transform infrared spectroscopy (FTIR) analysis revealed significant biochemical changes in the treated plants, indicating fungal activity. This study provides valuable insights into the efficacy of these BCAs and azoxystrobin, contributing to integrated management strategies for tomato leaf mold disease.

Application of Trichoderma Isolate consortium in organic fertilizer for controlling shallot twisted disease

The aim of the research was to determine the effect of applying a consortium of Trichoderma spp. isolates to a type of organic fertilizer that is effective in controlling twisted disease and on the growth and yield of shallot plants. The study utilized a completely randomized design (CRD) consisting of eight treatments with four replications. The treatments examined werea= as follows: P0 = control, P1 = 5 tons per ha of chicken manure fertilizer, P2 = 5 tons per ha of chicken manure fertilizer and consortium of two Trichoderma spp. isolates, P3 = 5 tons per ha of chicken manure fertilizer and consortium of three Trichoderma spp. isolates, P4 = 5 tons per ha of chicken manure fertilizer, 20 tons per ha trichocompost, and a consortium of two Trichoderma spp. isolates, P5 = 5 tons per ha of chicken manure fertilizer, 20 tons per ha trichocompost, and consortium of three Trichoderma spp. isolates, P6 = 20 tons per ha of trichocompost and a consortium of two Trichoderma spp. isolates, and P7 = 20 tons per ha of trichocompost and a consortium of three Trichoderma spp. isolates. The results showed that the twisted disease incidence in treatment P4 was the lowest (14.52%) at 8 weeks after planting. The plant height was of 39.10 cm, with 29.8 leaves per clump at 7 WAP, and the dried bulb weight was 112.4 g per clump. In conclusion, the application of chicken manure fertilizer at a dose of 5 ton per ha, combined with 20 tons per ha of trichocompost and a consortium of two Trichoderma spp. isolates, could control twisted disease in shallot.