Evaluating bioactivity of Trichoderma asperellum against Colletotrichum siamense and its growth-promoting effects on Aloe vera (Aloe barbadensis Mill.)

Evaluating bioactivity of Trichoderma asperellum against Colletotrichum siamense and its growth-promoting effects on Aloe vera (Aloe barbadensis Mill.) | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Evaluating bioactivity of Trichoderma asperellum against Colletotrichum siamense and its growth-promoting effects on Aloe vera (Aloe barbadensis Mill.) ANKUR MUKHOPADHYAY, Soumik Mukherjee, Subham Dutta, Sahely Kanthal, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5308483/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 18 Jan, 2025 Read the published version in European Journal of Plant Pathology → Version 1 posted 6 You are reading this latest preprint version Abstract Leaf spot disease caused by Colletotrichum siamense is a significant fungal threat to various plants, including Aloe vera. This study explores the biocontrol potential of Trichoderma asperellum against C. siamense while simultaneously evaluating the effects on Aloe vera growth parameters. Ten Trichoderma isolates (T 1 to T 10 ) were obtained from the rhizosphere of Aloe vera through serial dilution and assessed for their antagonistic activity using a dual culture technique. Among these isolates, five- T 1 , T 3 , T 4 , T 5 , and T 7 demonstrated the greatest suppression of radial growth of C. siamense , along with high sporulation rates. In pot tests, isolate T 3 emerged as particularly effective, enhancing plant weight by 144.30%, shoot length by 42.40%, shoot biomass by 144.40%, root length by 200%, root biomass by 146.20%, and leaf number by 20.80%. Additionally, T 3 significantly reduced the severity of leaf spot disease, achieving a 77.44% decrease in disease severity. Morphological and molecular characterization confirmed isolate T 3 as Trichoderma asperellum , with its internal transcribed spacer (ITS) sequence submitted to the NCBI GenBank and obtaining an accession number PP565067. These findings underscore the potential of T. asperellum as an effective biocontrol agent, promoting healthier growth in Aloe vera while simultaneously managing leaf spot disease, making it a promising solution for sustainable agriculture practices. Colletotrichum siamense Trichoderma asperellum serial dilution dual culture pot test ITS Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Trichoderma is predominantly soil-dwelling, cosmopolitan, asexual filamentous fungi (with teleomorphic forms known as Hypocrea ) that commonly inhabit decaying wood and other decomposing plant materials (Howell, 2003). These fungi are noted for their fast growth, featuring vibrant green conidia and conidiophores with a repetitively branched structure. Trichoderma belongs to the domain Eukaryota and is part of the kingdom Fungi, and categorised within the division Ascomycota, subdivision Pezizomycotina, class Sordariomycetes, order Hypocreales, and family Hypocreaceae. Hypocrea/Trichoderma encompasses over 300 species identified both molecularly and morphologically, with many still awaiting formal description (Bissett et al., 2015). Trichoderma employs multiple mechanisms to control phytopathogenic organisms such as competing with plant pathogens for resources, parasitizing and killing other fungi, producing antimicrobial substances, generating lytic enzymes, releasing secondary metabolites as well as induction of plant resistance (Vinale et al., 2008; Saravanakumar et al., 2016; Moosavi & Zare, 2020). Trichoderma strains are effective bio-pesticides and bio-fertilizers due to their direct competition with pathogens (Ghisalberti & Sivasithaparam, 1991). Additionally, applying Trichoderma strains to the soil and plants has been found to enhance both the productivity and quality of a variety of crops like cucumbers, tomatoes, carrots, beans, corn, cotton, tobacco, millet, and ornamental grasses which is likely due to the interaction between growth hormones produced by the fungi and the defence hormones triggered in the plant (Grant & Jones, 2009). It is recognized as an opportunistic plant symbiont and a potent mycoparasite. Trichoderma species compete with other pathogens for nutrients, growth factors, and space and also produce highly effective siderophores that sequester iron, thereby hindering the growth of other filamentous fungi (Benitez et al., 2004). Secondary metabolites produced by Trichoderma can be classified as volatile antibiotics, water-soluble compounds, and peptaibols, which are linear oligopeptides (Ghisalberti & Sivasithamparam, 1991). Trichoderma asperellum is widely regarded as a safe and effective biological control agent and does not persist in the food chain (Harman, 2006). These fungi can thrive in highly competitive environments and can counteract fungistatic effects (Benitez et al., 2004). Many strains exhibit resistance to a wide range of toxic substances, including metabolites produced by soil microorganisms or plants, as well as fungicides, herbicides, and antibiotics. T. asperellum has been shown to stimulate cucumber plants to produce indole-3-acetic acid (IAA), gibberellins (GA), and abscisic acid (ABA), which contribute to growth promotion (Liu et al., 2022). In tomato seedlings treated with T. asperellum , there were significant increases in plant height, stem diameter, soluble sugar content, and the absorption rate of available nitrogen. Additionally, the expression levels of hormone signalling-related genes, including JAR1, MYC2, NPR1, PR1, and GH3, were significantly elevated (Rawal et al., 2022). Another study indicated that T. asperellum can enhance the expression of xylanase genes in poplar, demonstrating a notable growth-promoting effect (Karuppiah et al., 2019). The genus Colletotrichum includes several significant plant pathogenic fungi responsible for anthracnose and postharvest rots affecting a variety of fruits, vegetables, and ornamental plants, particularly in tropical and subtropical areas (Hyde et al., 2009). In fact, Colletotrichum species are recognized among the top 10 fungal pathogens of both scientific and economic importance (Dean et al., 2012). Colletotrichum siamense has been reported from countries like China, the USA, Africa, Vietnam, and Thailand and is known to cause diseases in a wide range of plants (Prihastuti et al., 2009) including Aloe vera. By deploying Trichoderma , we can potentially mitigate the damage caused by C. siamense on Aloe vera, offering a sustainable alternative to chemical treatments while simultaneously enhancing various plant growth parameters. In this study, we assessed the effectiveness of Trichoderma asperellum against C. siamense and its impact on various plant growth parameters in Aloe vera. Materials and Methods Isolation of Colletotrichum siamense Colletotrichum siamense , the pathogen causing leaf spot in Aloe vera, was isolated in February 2022 from infected leaves at the Medicinal Plants Garden (22°59'21"N latitude and 88°27'18"E longitude) under the All India Coordinated Research Project on Medicinal and Aromatic Plants and Betel Vine, Directorate of Research, Bidhan Chandra Krishi Viswavidyalaya, Nadia, West Bengal, India. The pathogen is present year-round, peaking in winter. To isolate it, infected leaf pieces underwent surface sterilization with 0.1% mercuric chloride, followed by rinsing with sterile water, and cultured on Potato Dextrose Agar (PDA) with streptomycin sulphate. After 4–5 days of incubation at 27 ± 1°C, single hyphal tips were sub-cultured to establish a pure culture and stored at 4°C. Identification of C. siamense was confirmed through pathogenicity tests and molecular analysis of ITS, β-tubulin, and glyceraldehyde-3-phosphate dehydrogenase regions, with accession numbers OQ423208, PQ285628, and PQ285629 respectively. Isolation of Trichoderma spp. During February 2022, soil samples were randomly collected from a depth of 5 to 15 cm around the rhizosphere of Aloe vera plants at the Medicinal Plants Garden (22°59'21"N latitude and 88°27'18"E longitude) under the All India Coordinated Research Project on Medicinal and Aromatic Plants and Betel Vine, Directorate of Research, Bidhan Chandra Krishi Viswavidyalaya, Nadia, West Bengal, India. Trichoderma species were isolated using serial dilution technique. For this purpose, Trichoderma -specific media was prepared using the following composition: MgSO₄·7H₂O (0.2 g), K₂HPO₄ (0.9 g), KCl (0.15 g), NH₄NO₃ (3.0 g), Glucose (3.0 g), Agar (15 g), Rose Bengal (0.15 g), Chloramphenicol (0.25 g), and distilled water to a final volume of 1000 ml, adjusted to a pH of 6.5 (Elad et al., 1981). The initial dilution was created by adding 1 g of the representative soil sample to 9 ml of sterilized water in the first test tube, mixed thoroughly, and labelled as 10 − 1 . Following this, 1 ml of the suspension from the first dilution was transferred to a second tube containing 9 ml of sterilized water, resulting in the 10 − 2 dilution. This process was repeated until the original sample was diluted to 10 − 6 . From this 10 − 6 dilution, 1 ml was spread onto Petri dishes containing the Trichoderma -specific media, supplemented with streptomycin sulphate (0.01%) as an antibiotic. The inoculated plates were then incubated in a BOD incubator at 27 ± 1°C for 4–5 days to monitor fungal colony growth. After the incubation period, individual colonies exhibiting yellowish-green and whitish-green pigmentation were identified and transferred to sterilized Potato Dextrose Agar plates using a sterile loop to obtain pure cultures, which were subsequently stored in a refrigerator at 4°C for further analysis (Arumugam et al., 2013). Identification of Trichoderma isolates was based on cultural and microscopic characteristics. Colony morphology was assessed on PDA, and the lactophenol cotton blue staining technique was used for microscopic examination of conidiophores, conidia, and phialides, facilitating accurate identification of the isolates. Dual culture technique The antagonistic potential of the isolated Trichoderma spp. was assessed against C. siamense using dual culture technique (Ruocco et al., 2009). In this method, a 5 mm mycelial disc from 7 days old cultures of both the pathogen and the biocontrol agents was placed at opposite peripheries of 9 cm Petri plates containing PDA and the dual culture plates were incubated at 27 ± 1°C. Plates inoculated with only the disc of test pathogen served as control. There were three replications for each treatment. Antagonistic activity was evaluated by measuring the radial growth of the pathogen and comparing it with the growth observed in the control plates. Measurements of colony diameter for both the pathogen and the biocontrol agents were recorded after 3 and 7 days of incubation allowing for a comprehensive assessment of the interaction between the biocontrol agent and the pathogen. The percentage inhibition of radial growth (PIRG) of the pathogen was estimated by the formula given below (Vincent, 1947) PIRG = (C - T / C) × 100 Where, PIRG = Percentage inhibition of radial growth, C = Radial growth of pathogen in control and T = Radial growth of pathogen in presence of antagonist. The degree of antagonism between the biocontrol agent and test pathogen in dual culture (Table 1 ) was scored on a scale of 1–5 (Bell et al., 1982). Table 1 Bell’s scale for estimating the degree of antagonism in dual culture test Scale Degree of antagonisms 1 = The antagonist fully outgrew the pathogen, covering the entire surface of the medium. 2 = The antagonist covered at least two-thirds of the medium's surface. 3 = Both the antagonist and the pathogen occupied roughly half of the medium's surface (more than one-third but less than two-thirds), with neither organism showing dominance over the other. 4 = The pathogen occupied at least two-thirds of the medium's surface and seemed to resist encroachment. 5 = The pathogen fully outgrew the antagonist, covering the entire surface of the medium. Assessing sporulation capacity of Trichoderma via haemocytometer for enhancing Aloe vera growth in Pot Culture Selecting Trichoderma isolates based on their sporulation capacity is essential for enhancing the growth and health of Aloe vera in pot culture, as this influences the development of effective bioformulations (Jeyarajan & Nakkeeran, 2000). Isolates are cultured on media like Potato Dextrose Agar (PDA) for 7 days, with sporulation assessed through visual inspection and spore quantification using a haemocytometer. Spores are collected by adding sterile distilled water to the cultures and agitating gently. If the concentration is high, serial dilutions may be performed to ensure an appropriate count. A small volume of the spore suspension (typically 10 µL) is loaded onto the haemocytometer, covered with a coverslip, and observed under a light microscope where spore concentration (spores/mL) is determined using a formula that accounts for the total number of spores counted, the dilution factor, and the volume counted (Kamaruzzaman et al., 2016). High sporulation rates are prioritized, as these isolates are more likely to establish robust populations and effectively colonize the root zone of Aloe vera. The most promising isolates are then applied in pot culture experiments to evaluate their effects on plant growth, disease resistance, and overall health. Evaluation of Trichoderma Isolates for efficacy on Aloe vera in-vivo Selected Trichoderma isolates were assessed for their effectiveness on Aloe vera plants grown in pots within a shade net house. The potting medium consisted of garden soil mixed with farmyard manure (FYM) in a 50:50 v/v ratio, which was used to cultivate young Aloe vera plants at 4–5 leaf stage. Each Trichoderma isolate was cultured in potato dextrose broth at 27 ± 1°C for 7 days. A talc-based bioformulation containing approximately 1×10 8 spores per gram was prepared using spores harvested from the mycelial mat of the broth culture (Boblina et al., 2020). Each bioformulation was incorporated into the potting soil at a rate of 5 grams per kilogram (Kumar et al., 2014). Subsequently, Aloe vera seedlings were transplanted into the pots after soaking their roots in a solution mixed with the bioformulation @ 5 grams per litre of water. Each treatment included four replicates, while a control group was maintained separately. Plants were watered as necessary to ensure optimal moisture levels. Over the course of 120 days, the plants were monitored, and data were collected on disease symptoms, plant height, shoot and root biomass, and the number of leaves. Identification of Trichoderma asperellum Identification of Trichoderma asperellum as an effective bio-control agent against Colletotrichum siamense and a growth promoter for Aloe vera was achieved through a combination of morphological assessments and molecular techniques. Key morphological characteristics, including colony colour, texture, and the structure of conidiophores, conidia, and phialides, were examined under stereo microscopes [Nikon (Model- SMZ 25)] and light microscopes [Nikon Trinocular Research Microscope (Model: Ci-L)]. To confirm the molecular identity, genomic DNA was extracted using the CTAB method (Doyle & Doyle, 1987). The amplification of the Internal Transcribed Spacer (ITS) region was conducted with the ITS1 and ITS4 primers. (White et al., 1990). Polymerase Chain Reactions (PCR) were conducted in a 25 µL reaction volume using the Veriti™ PCR system. Each reaction included 2 µL of DNA sample, 1 µL each of forward and reverse primers, 12.5 µL of 2x Taq PCR Mix, and 8.5 µL of PCR grade water (GCC Biotech, India). The PCR protocol involved an initial denaturation at 95°C for 5 minutes, followed by 35 cycles of denaturation at 95°C for 1 minute, annealing at 60°C for 1 minute, and extension at 72°C for 1.5 minutes, concluding with a final elongation at 72°C for 8 minutes (White et al., 1990). The amplified PCR products were examined through electrophoresis on a 1.5% agarose gel, using a 100 bp DNA ladder for size determination (GCC Biotech, India). The PCR products were purified and sequenced using the Sanger method, with the resulting sequences compared to the NCBI GenBank database (Altschul et al., 1990) to confirm species identification and assess phylogenetic relationships. Further evolutionary analysis using MEGA X (Kumar et al., 2018) supported the identification of T. asperellum . Results Isolation of Trichoderma isolates A total of ten Trichoderma species were isolated from the rhizosphere of Aloe vera using Trichoderma selective medium (TSM) and subsequently subcultured on Potato Dextrose Agar (PDA). The isolates were designated as T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , and T 10 based on their morphological characteristics (Fig. 1 ). To assess their biocontrol potential, the ability of these Trichoderma isolates to inhibit the mycelial growth of Colletotrichum siamense was evaluated through dual culture experiments on PDA medium. Effect of Trichoderma species on mycelial growth of Colletotrichum siamense in vitro All ten Trichoderma isolates demonstrated an antagonistic effect against Colletotrichum siamense , effectively inhibiting its mycelial growth. After three days of inoculation, isolate T 3 exhibited the strongest antagonism, achieving a maximum reduction in colony growth to 2.03 cm, which corresponds to a 61.85% reduction. This was closely followed by isolate T 2 , which reduced the growth to 2.13 cm (59.97%), with both isolates showing statistically similar effects. The least inhibitory effect was noted in isolate T 5 , which achieved a 54.97% reduction. By the seventh day post-inoculation, isolates T 3 and T 7 recorded the highest reduction at 68.05%, with isolate T 4 at 67.63%, and isolates T 1 , T 2 , and T 10 each at 67.21%, all of which were statistically comparable. Isolate T 6 showed the lowest inhibition percentage at 60.07% (Table 2 ; Fig. 2 ). Additionally, the presence of the Trichoderma isolates resulted in observable changes in the morphological features of the pathogen (Table 3 ). Table 2 Test of efficacy of different Trichoderma isolates against Colletotrichum siamense in dual culture test on PDA incubated at 27 ± 1°C Radial Growth of Pathogen (cm) Isolates 3rd Day* Inhibition (%) 7th Day* Inhibition (%) Bell’s Scale (on 7th day) Sporulation (x10 6 spores/ sq.cm) T 1 2.20 58.72 2.60 67.21 1 2.20 T 2 2.13 59.97 2.60 67.21 1 1.10 T 3 2.03 61.85 2.53 68.05 1 2.80 T 4 2.20 58.72 2.57 67.63 1 2.10 T 5 2.40 54.97 2.63 66.79 1 2.30 T 6 2.20 58.72 3.17 60.07 1 1.80 T 7 2.20 58.72 2.53 68.05 1 2.60 T 8 2.23 58.10 2.97 62.59 1 0.50 T 9 2.23 58.10 2.70 65.95 1 1.20 T 10 2.23 58.10 2.60 67.21 1 1.40 Control 5.33 - 7.93 - - - S. Em± 0.07 0.05 0.07 C. D. (5%) 0.15 0.10 0.14 C.V. (%) 3.63 1.98 4.65 *Average of three replications. Inhibition (%) was calculated over control Some Trichoderma isolates induced noticeable morphological changes in Colletotrichum siamense , particularly concerning colony colour and topography, compared to the control plate. The pure culture of the pathogen typically displayed a cottony, pink-white aerial mycelium with a flat topography. In contrast, isolate T 2 produced a cottony mycelium that was yellow to yellow-brown with a fluffy, raised appearance. Both isolates T 7 and T 9 exhibited yellow to brownish cottony mycelium with a similarly fluffy, elevated topography. Isolate T 10 , on the other hand, formed a cottony, dirty white mycelium, characterized by yellow margins and a fluffy, raised structure. These morphological alterations highlight the impact of Trichoderma isolates on the pathogen's growth characteristics (Fig. 2 ). Table 3 Influence of Trichoderma isolates on the growth behaviour of Colletotrichum siamense evaluated in dual culture on PDA, incubated at 27 ± 1°C for 7 days. Trichoderma isolates Colony Colour Front plate Colony Colour Reverse plate Topography T 1 Cottony, white to off-white mycelium Cottony, white to off-white mycelium Flat T 2 Cottony, yellow to yellow-brown mycelium Cottony, yellow to yellow-brown mycelium Fluffy, raised T 3 Aerial, white mycelium Aerial, white mycelium Flat T 4 Cottony, aerial, white mycelium Cottony, aerial, white mycelium Flat T 5 Off-white to grey aerial mycelium Off-white to grey aerial mycelium Fluffy, raised T 6 Cottony, pink-white, aerial mycelium Cottony, pink-white, aerial mycelium Flat T 7 Yellow to brownish cottony mycelium Yellow to brownish cottony mycelium Fluffy, raised T 8 Pink-white cottony aerial mycelium Pink-white cottony aerial mycelium Flat T 9 Cottony, yellow to brownish mycelium Cottony, yellow to brownish mycelium Fluffy, raised T 10 Cottony, dirty white to yellowish colour at the margin Cottony, dirty white to yellowish colour at the margin Fluffy, raised Control Cottony, Pink-white aerial mycelium Cottony, Pink-white aerial mycelium Flat The results from the in vitro dual culture test indicated that Trichoderma isolate T 3 exhibited the highest inhibition of colony growth of Colletotrichum siamense , achieving a reduction of 61.85%. This finding underscores the potential of isolate T 3 as an effective biocontrol agent against this pathogen. Spore producing capacity of Trichoderma spp. In this study, the spore production capacity of the ten Trichoderma spp. after a 7-day incubation period on PDA at 27 ± 1°C was observed. It was found that isolate T 3 exhibited highest spore concentration, with 2.8x10 6 spores per square centimetre of Petri plate. This was followed closely by isolates T 7 , T 5 , T 1 , and T 4 , which produced 2.6x10 6 , 2.3x10 6 , 2.2x10 6 , and 2.1x10 6 spores per square centimetre, respectively. In contrast, isolate T 8 demonstrated the lowest spore concentration, with only 0.5x10 6 spores per square centimetre (Fig. 3 ). In dual culture tests, all ten Trichoderma isolates demonstrated effectiveness in suppressing the growth of Colletotrichum siamense in vitro . Among these isolates, five- T 1 , T 3 , T 4 , T 5 , and T 7 exhibited significantly higher sporulation capacities, producing greater numbers of spores compared to the other five isolates. Due to their superior performance in sporulation, these five isolates were selected for further evaluation in pot tests. Performance of selected Trichoderma isolates in reducing leaf spot disease and their effects on Aloe Vera in vivo After 120 days, Trichoderma inoculation significantly enhanced all growth metrics compared to the control (un-inoculated plants). Additionally, the inoculated plants exhibited reduced disease severity, indicating the protective role of Trichoderma against the target pathogen. Isolate T 3 exhibited the highest plant weight at 243.90 g, reflecting a 144.30% increase over control, while isolate T 4 showed only a 15.50% increase with 115.33 g. T 3 also produced the longest shoot length (32.70 cm, 42.40% increase) and the greatest shoot biomass (228.17 g, 144.40% increase). In terms of root growth, T 3 had the longest root length (25.09 cm, 200% increase) and highest root biomass (15.98 g, 146.20% increase). Additionally, T 3 showed a 20.80% increase in the number of leaves, while T 1 and T 5 had no significant effect. Overall, isolate T 3 demonstrated superior performance in enhancing growth parameters, indicating its potential as an effective inoculant for Aloe vera (Table 4 ; Fig. 4 ). Table 4 Impact of Trichoderma isolates on various growth parameters of Aloe vera after 120 days under pot culture experiment Trichoderma i solates No. of leaves Increase (%) Length of Shoot (cm) Increase (%) Biomass of shoot (gm) Increase (%) Length of root (cm) Increase (%) Biomass of root (gm) Increase (%) Plant weight (gm) Increase (%) T 1 6.00 0.00 28.08 22.30 111.59 19.50 12.48 49.30 8.49 30.80 120.33 20.50 T 3 7.25 20.80 32.70 42.40 228.17 144.40 25.09 200.00 15.98 146.20 243.90 144.30 T 4 7.00 16.70 26.34 14.70 108.10 15.80 10.36 23.90 7.24 11.50 115.33 15.50 T 5 6.00 0.00 29.96 30.40 163.27 74.90 15.23 82.10 9.24 42.30 172.25 72.50 T 7 6.25 4.20 31.70 38.00 212.95 128.10 16.48 97.00 13.73 111.50 226.68 127.00 Control 6.00 - 22.97 - 93.37 - 8.36 - 6.49 - 99.86 - S. Em± 0.39 1.79 6.27 0.95 0.78 12.15 C. D. (5%) 0.82 3.76 13.17 1.99 1.64 25.53 C. V. (%) 8.61 8.83 5.80 9.01 10.88 10.54 * Data were calculated per plant with four replications, and the percentage increase was determined in comparison to the control Table 5 Effect of Trichoderma isolates on Percent Disease Index (PDI) and Disease Incidence (%) with corresponding decrease (%) after 120 Days in pot culture experiment Trichoderma isolates Leaf spot caused by Colletotrichum siamense PDI Decrease (%) Incidence (%) Decrease (%) T 1 3.83 (11.28) 73.10 19.17 (25.95) 73.10 T 3 3.21 (10.32) 77.44 16.07 (23.62) 77.44 T 4 3.33 (10.52) 76.61 16.67 (24.09) 76.61 T 5 3.83 (11.28) 73.10 19.17 (25.95) 73.10 T 7 4.00 (11.54) 71.93 20.00 (26.57) 71.93 Control 14.25 (22.16) - 71.25 (57.69) S. Em± 0.43 1.45 C. D. (5%) 0.91 3.05 C. V. (%) 4.77 6.69 *Data were calculated per plant with four replications. The percentage decrease was determined relative to the control, with values in parentheses indicating angular transformation (arc sin) . For Colletotrichum siamense , isolate T 3 exhibited the highest reduction in both PDI and disease incidence, achieving a decrease of 77.44%. In contrast, isolate T 7 showed the least reduction at 71.93% (Table 5 ). Overall, isolate T 3 had a beneficial effect on Aloe vera plants, significantly lowering the PDI and disease incidence for the target pathogen. In conclusion, the findings from both the in vitro dual culture tests and the in vivo pot tests indicate that Trichoderma isolate T 3 effectively suppressed the growth of Colletotrichum siamense . Additionally, isolate T 3 demonstrated the highest capacity for sporulation and promoted plant growth while inducing resistance in Aloe vera plants. Identification of Trichoderma isolate T 3 Trichoderma isolate T 3 completely covered the Potato Dextrose Agar (PDA) plate by the fourth day when incubated at 27 ± 1°C. Initially, the colonies appeared white, but the mycelium became pale green to dark green after a few days. The conidiophores were branched and produced in pustules, ending in either a single phialide or a cluster of two to three diverging phialides (Fig. 5 ). The conidia (N = 50) were dark green, globose to sub-globose, measuring 3.30 ± 1.20 µm in length (range: 2.50–4.60 µm) and 3.02 ± 0.95 µm in breadth (range: 2.15–3.90 µm). The internal transcribed spacer (ITS) region was amplified and sequenced to ensure accurate species identification. BLASTn searches conducted in the NCBI database indicated that the ITS sequence of the isolate exhibited a high degree of similarity to other T. asperellum strains, with identity percentages ranging from 99.84–100%. Following the submission of the nucleotide sequences to the NCBI GenBank database, the accession number assigned was PP565067. The phylogenetic relationship of the isolate was analyzed using the Jukes-Cantor model within the maximum likelihood method, implemented in MEGA X software (Fig. 6 ). Discussion Numerous studies have investigated the biological control of Colletotrichum species using various strains of Trichoderma . In dual culture experiments, it was observed that hyphal inhibition commenced upon contact between the antagonist and the pathogen (Almeida et al., 2007). Trichoderma spp. effectively suppressed the hyphal growth of Colletotrichum spp. through mechanisms such as hyphal coiling, the induction of hyphal abnormalities, reduction in acervuli production, and lysis of both hyphae and sclerotia (Malathi et al., 2002). For instance, Trichoderma viride demonstrated significant effectiveness, achieving 70.42% inhibition of Colletotrichum gloeosporioides (Patil et al., 2009). T. asperellum strain T8a led to 80% growth inhibition of C. gloeosporioides and an 85.18% inhibition of C. asianum (De los Santos-Villalobos et al., 2013). Additionally, T. asperellum isolate 21 also exhibited effective results in dual culture against these pathogens (Sharma & Prasad, 2018). Similarly, T. longibrachiatum and T. harzianum showed inhibition rates of 55.82% and 53.26% against C. gloeosporioides , respectively (Valenzuela et al., 2015). Notably, isolates of T. harzianum were found to be particularly effective, reducing the growth of C. gloeosporioides by 42% (Prabakar et al., 2008). Certain strains of Trichoderma are known for their ability to establish robust and long-lasting colonization on root surfaces by penetrating the root epidermis. This process enhances root growth and improves resistance to abiotic stresses through better mineral absorption (Harman, 2006). The observed improvements in various plant growth parameters in the present study align with findings from several researchers (Chang et al., 1986; Ahmad & Baker, 1987; Yaqub & Shahzad, 2011). Additionally, it has been reported that plants inoculated with Trichoderma exhibit increased disease resistance in plants, attributed to higher production of defence-related enzymes, including peroxidase, phenylalanine ammonia-lyase, and β-1,3-glucanase. These inoculations also contribute to earlier emergence and enhanced plant vigour (Jogaiah et al., 2013). When Trichoderma asperellum was applied to the aerial parts of strawberry plants infected with anthracnose caused by Colletotrichum , a significant reduction in both disease incidence and severity was observed compared to untreated plants under greenhouse conditions (Kaissoumi et al., 2022). Declarations Acknowledgments: This work was supported by the Indian Council of Agricultural Research - All India Coordinated Research Project (AICRP) on Medicinal and Aromatic Plants and Betel vine, Directorate of Research, Bidhan Chandra Krishi Viswavidyalaya, Kalyani-741235, West Bengal, India, and National Medicinal Plant Board, Ministry of AYUSH, Government of India. Ethical Approval: Ethical approval was not applicable for this study since it exclusively involved using plant and microflora samples, which typically do not require human or animal ethics review. Author contribution: All authors have contributed to the manuscript's study, conception, and design. Material preparation, data collection, and analysis- Ankur Mukhopadhyay, Soumik Mukherjee, Subham Dutta, and Goutam Mondal. Conceptualization- Ankur Mukhopadhyay and Goutam Mondal; Methodology- Ankur Mukhopadhyay, Soumik Mukherjee, and Subham Dutta. Writing and original draft preparation- Ankur Mukhopadhyay; Formal analysis and investigation- Ankur Mukhopadhyay and Goutam Mondal. Review and editing- Soumik Mukherjee, Subham Dutta, and Sahely Kanthal; Supervision- Goutam Mondal. Conflict of interest: The authors declare no conflict of interest. Financial interest: The authors have no relevant financial or non-financial interests to disclose. Data availability : The sequence data generated for this study has been submitted to the NCBI GenBank database with accession number PP565067 for ITS. References Ahmad, J. S., & Baker, R. (1987). Rhizosphere competence of Trichoderma harzianum . Phytopathology , 77 (2), 182–189. https://doi.org/10.1094/Phyto-77-182 Almeida, F. B. D. R., Cerqueira, F. M., Silva, R. D. N., Ulhoa, C. J., & Lima, A. L. (2007). Mycoparasitism studies of Trichoderma harzianum strains against Rhizoctonia solani : evaluation of coiling and hydrolytic enzyme production. Biotechnology letters , 29, 1189–1193. https://doi.org/10.1007/s10529-007-9372-z Altschul, S. F., Gish, W., Miller, W., Myers, E. W., & Lipman, D. J. (1990). Basic local alignment search tool. Journal of molecular biology , 215(3), 403–410. https://doi.org/10.1016/s0022-2836(05)80360-2 Arumugam, K., Ramalingam, P., & Appu, M. (2013). Isolation of Trichoderma viride and Pseudomonas fluorescens organism from soil and their treatment against rice pathogens. Journal of Microbiology and Biotechnology Research , 3 (6),77–81. Azad, R., Hamza, A., Khatun, M. M., Nasrin, S., Tanny, T., Das, K. C., & Alam, I. (2020). First report of Colletotrichum siamense causing leaf spot in Aloe vera in Bangladesh. Plant Disease , 104 (11), 3058. https://doi.org/10.1094/PDIS-04-20-0857-PDN Bell, D. K., Wells, H. D., & Markham, C. R. (1982). In vitro antagonism of Trichoderma species against six fungal plant pathogens. Phytopathology , 72 , 379–382. https://doi.org/10.1094/Phyto-72-379 Benítez, T., Rincón, A. M., Limón, M. C., & Codón, A. C. (2004). Biocontrol mechanisms of Trichoderma strains. International Microbiology , 7 (4), 249–260. Bissett, J., Gams, W., Jaklitsch, W., & Samuels, G. J. (2015). Accepted Trichoderma names in the year 2015. IMA fungus , 6 , 263–295. https://doi.org/10.5598/imafungus.2015.06.02.02 Boblina, B., Beura, S. K., Panda, A. G., & Mishra, M. K. (2020). Evaluation and assessment of shelf life of liquid substrates and talc formulation for mass production of native Trichoderma spp. Journal of Pharmacognosy and Phytochemistry , 9(3), 911–915. Chang, Y. C., Baker, R., Kleifeld, O., & Chet, I. (1986). Increased growth of plants in the presence of the biological control agent Trichoderma harzianum . Plant Disease , 70 (2):145–148. https://doi.org/10.1094/PD-70-145 De los Santos-Villalobos, S., Guzmán-Ortiz, D. A., Gómez-Lim, M. A., Délano-Frier, J. P., de-Folter, S., Sánchez-García, P., & Peña-Cabriales, J. J. (2013). Potential use of Trichoderma asperellum (Samuels, Liechfeldt et Nirenberg) T8a as a biological control agent against anthracnose in mango ( Mangifera indica L.). Biological control , 64 (1), 37–44. https://doi.org/10.1016/j.biocontrol.2012.10.006 Dean, R., Van Kan, J. A., Pretorius, Z. A., Hammond‐Kosack, K. E., Di Pietro, A., Spanu, P. D., & Foster, G. D. (2012). The Top 10 fungal pathogens in molecular plant pathology. Molecular plant pathology , 13 (4), 414–430. https://doi.org/10.1111/j.1364-3703.2011.00783.x Doyle, J. J., & Doyle, J. L. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical bulletin , 19 , 11–15. Elad, Y., Chet, I., & Henis, Y. (1981). A selective medium for improving quantitative isolation of Trichoderma spp. from soil. Phytoparasitica , 9 , 59–67. https://doi.org/10.1007/BF03158330 Ghisalberti, E. L., & Sivasithamparam, K. (1991). Antifungal antibiotics produced by Trichoderma spp. Soil biology and Biochemistry , 23 (11), 1011–1020. https://doi.org/10.1016/0038-0717(91)90036-J Grant, M. R., & Jones, J. D. (2009). Hormone (dis) harmony moulds plant health and disease. Science , 324 (5928), 750–752. https://doi.org/10.1126/science.1173771 Harman, G. E. (2006). Overview of Mechanisms and Uses of Trichoderma spp. Phytopathology , 96 (2), 190–194. https://doi.org/10.1094/PHYTO-96-0190 Howell, C. R. (2003). Mechanisms employed by Trichoderma species in the biological control of plant diseases: the history and evolution of current concepts. Plant disease , 87 (1), 4–10. https://doi.org/10.1094/PDIS.2003.87.1.4 Hyde, K. D., Cai, L., Cannon, P. F., Crouch, J. A., Crous, P. W., Damm, U., & Tan, Y. P. (2009). Colletotrichum - names in current use. Fungal Diversity , 39 (1), 147–182. Jeyarajan, R., & Nakkeeran, S. (2000). Exploitation of microorganisms and viruses as biocontrol agents for crop disease management. In Biocontrol Potential and its Exploitation in Sustainable Agriculture: Crop Diseases, Weeds, and Nematodes (pp. 95-116). Boston, MA: Springer US. https://doi.org/10.1007/978-1-4615-4209-4_8 Jogaiah, S., Abdelrahman, M., Tran, L. S. P., & Shin-Ichi, I. (2013). Characterization of rhizosphere fungi that mediate resistance in tomato against bacterial wilt disease. Journal of experimental botany , 64 (12), 3829–3842. https://doi.org/10.1093/jxb/ert212 Kaissoumi, H., Berber, F., Mouden, N., Ouazzani Chahdi, A., Albatnan, A., Ouazzani Touhami, A., & Douira, A. (2022). Effect of Trichoderma asperellum on the development of strawberry plants and biocontrol of anthracnose disease caused by Colletotrichum gloeosporioides . In International Conference on Advanced Intelligent Systems for Sustainable Development (pp. 609-622). Cham: Springer Nature Switzerland. https://doi.org/10.1007/978-3-031-35248-5_55 Kamaruzzaman, M., Rahman, M. M., Islam, M. S., & Ahmad, M. U. (2016). Efficacy of four selective Trichoderma isolates as plant growth promoters in two peanut varieties. International Journal of Biological Research , 4 (2), 152–156. https://doi.org/10.14419/ijbr.v4i2.6468 Karuppiah, V., Vallikkannu, M., Li, T., & Chen, J. (2019). Simultaneous and sequential based co-fermentations of Trichoderma asperellum GDFS1009 and Bacillus amyloliquefaciens 1841: a strategy to enhance the gene expression and metabolites to improve the bio-control and plant growth promoting activity. Microbial Cell Factories , 18 , 1–16. https://doi.org/10.1186/s12934-019-1233-7 Kimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of molecular evolution , 16 , 111–120. https://doi.org/10.1007/BF01731581 Kumar, S., Stecher, G., Li, M., Knyaz, C., & Tamura, K. (2018). MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular biology and evolution , 35 (6), 1547–1549. https://doi.org/10.1093/molbev/msy096 Kumar, S., Thakur, M., & Rani, A. (2014). Trichoderma : Mass production, formulation, quality control, delivery and its scope in commercialization in India for the management of plant diseases. African journal of agricultural research , 9 (53), 3838–3852. https://doi.org/10.5897/AJAR2014. 9061 Liu, Y., He, P., He, P., Munir, S., Ahmed, A., Wu, Y., & He, Y. (2022). Potential biocontrol efficiency of Trichoderma species against oomycete pathogens. Frontiers in Microbiology , 13 , 974024. https://doi.org/10.3389/fmicb.2022.974024 Malathi, P., Viswanathan, R., Padmanaban, P., Mohanraj, D., & Sunder, A. R. (2002). Compatibility of biocontrol agents with fungicides against red rot disease of sugarcane. Sugar Tech , 4 , 131–136. https://doi.org/10.1007/BF02942694 Moosavi, M. R., & Zare, R. (2020). Fungi as biological control agents of plant-parasitic nematodes. Plant defence: biological control , 333–384. https://doi.org/10.1007/978-3-030-51034-3_14 Patil, C. U., Zape, A. S., & Wathore, S. D. (2009). Efficacy of fungicides and bioagents against Colletotrichum gloeosporioides causing blight in Piper longum . International Journal of Plant Protection , 2 (1), 63–66. Prabakar, K., Raguchander, T., Saravanakumar, D., Muthulakshmi, P., Parthiban, V. K., & Prakasam, V. (2008). Management of postharvest disease of mango anthracnose incited by Colletotrichum gleosporioides . Archives of Phytopathology and Plant protection , 41 (5), 333–339. https://doi.org/10.1080/03235400600793502 Prihastuti, H., Cai, L., Chen, H., McKenzie, E. H. C., & Hyde, K. D. (2009). Characterization of Colletotrichum species associated with coffee berries in northern Thailand. Fungal Diversity , 39 (1), 89–109. Rawal, R., Scheerens, J. C., Fenstemaker, S. M., Francis, D. M., Miller, S. A., & Benitez, M. S. (2022). Novel Trichoderma isolates alleviate water deficit stress in susceptible tomato genotypes. Frontiers in plant science , 13 , 869090. https://doi.org/10.3389/fpls.2022.869090 Ruocco, M., Lanzuise, S., Vinale, F., Marra, R., Turrà, D., Woo, S. L., & Lorito, M. (2009). Identification of a new biocontrol gene in Trichoderma atroviride : the role of an ABC transporter membrane pump in the interaction with different plant-pathogenic fungi. Molecular plant-microbe interactions , 22 (3), 291–301. https://doi.org/10.1094/MPMI-22-3-0291 Saravanakumar, K., Yu, C., Dou, K., Wang, M., Li, Y., & Chen, J. (2016). Synergistic effect of Trichoderma -derived antifungal metabolites and cell wall degrading enzymes on enhanced biocontrol of Fusarium oxysporum f. sp. cucumerinum . Biological control , 94 , 37–46. https://doi.org/10.1016/j.biocontrol.2015.12.001 Sharma, K. S., & Prasad, L. (2018). Bioactivity of Trichoderma asperellum against Colletotrichum asianum and Sclerotinia sclerotiorum . Pesticide Research Journal , 30 (2), 251–255. https://doi.org/ 10.5958/2249-524X.2018.00040.7 Valenzuela, N. L., Angel, D. N., Ortiz, D. T., Rosas, R. A., García, C. F. O., & Santos, M. O. (2015). Biological control of anthracnose by postharvest application of Trichoderma spp. on maradol papaya fruit. Biological Control , 91 , 88–93. https://doi.org/10.1016/j.biocontrol.2015.08.002 Vinale, F., Marra, R., Scala, F., Ghisalberti, E. L., Lorito, M., & Sivasithamparam, K. (2006). Major secondary metabolites produced by two commercial Trichoderma strains active against different phytopathogens. Letters in applied microbiology , 43 (2), 143–148. https://doi.org/10.1111/j.1472-765X.2006.01939.x Vincent, J. M. (1947). Distortion of fungal hyphae in the presence of certain inhibitors. Nature , 159 (4051), 850–850. https://doi.org/10.1038/159850b0 White, T. J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR protocols a guide to methods and applications, 315–322. Yaqub, F., & Shahzad, S. (2011). Efficacy and persistence of micobial antagonists against Sclerotium rolfsii under field conditions. Pakistan Journal of Botany , 43 (5), 2627–2634. Cite Share Download PDF Status: Published Journal Publication published 18 Jan, 2025 Read the published version in European Journal of Plant Pathology → Version 1 posted Editorial decision: Revision 02 Dec, 2024 Reviewers agreed at journal 31 Oct, 2024 Reviewers invited by journal 31 Oct, 2024 Editor invited by journal 31 Oct, 2024 Editor assigned by journal 30 Oct, 2024 First submitted to journal 21 Oct, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5308483","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":372607702,"identity":"e627deb2-fcae-4406-b191-d11137721621","order_by":0,"name":"ANKUR MUKHOPADHYAY","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABE0lEQVRIiWNgGAWjYFACNjYQAWIxA7GEHD+ImVBAvBYbY8kGkBYDgloYYFrSEjccALHxaNGdkZb24OMePjl+9rOHjXn3HDY2Pr868cMDAwZ5frEDWLWY3Ug7bjjjGZuxZE9ecjLPs8NyZjfebpYAOsxw5uwEHFrS26R5DrAB3ZNjfJjnwGFjsxtnN4C0JBjcxqPlzwG2+v3n34C1JG6ecXbzD/xa0o5JMxxgSzCQyDFO5jkA9D5/7zb8tpx5libZc4DNcMaNN8aGcw7YGEvc4N1mATQBt1+Op5lJ/DhwTJ6/P8dY4s0BYFT2n91880eFjTy/NHYtUHAMiS0BVimBTzkI1CCx+Q8QUj0KRsEoGAUjDAAA5GZi6AvlbRcAAAAASUVORK5CYII=","orcid":"https://orcid.org/0009-0008-5163-1415","institution":"Brainware University","correspondingAuthor":true,"prefix":"","firstName":"ANKUR","middleName":"","lastName":"MUKHOPADHYAY","suffix":""},{"id":372607703,"identity":"91740d4d-2f0f-4d1d-a44a-b7ce0f63296a","order_by":1,"name":"Soumik Mukherjee","email":"","orcid":"","institution":"Bidhan Chandra Krishi Viswavidyalaya: Bidhan Chandra Krishi Viswa Vidyalaya","correspondingAuthor":false,"prefix":"","firstName":"Soumik","middleName":"","lastName":"Mukherjee","suffix":""},{"id":372607704,"identity":"31ef1281-4c21-4500-b15f-d3ed28df46cd","order_by":2,"name":"Subham Dutta","email":"","orcid":"","institution":"Bidhan Chandra Krishi Viswavidyalaya: Bidhan Chandra Krishi Viswa Vidyalaya","correspondingAuthor":false,"prefix":"","firstName":"Subham","middleName":"","lastName":"Dutta","suffix":""},{"id":372607705,"identity":"af53be2e-2ae7-421f-8164-8bd154cb55a8","order_by":3,"name":"Sahely Kanthal","email":"","orcid":"","institution":"Brainware University","correspondingAuthor":false,"prefix":"","firstName":"Sahely","middleName":"","lastName":"Kanthal","suffix":""},{"id":372607706,"identity":"5d652daa-895f-4db8-bd8b-ac07edb0e3f5","order_by":4,"name":"Goutam Mondal","email":"","orcid":"","institution":"Bidhan Chandra Krishi Viswavidyalaya: Bidhan Chandra Krishi Viswa Vidyalaya","correspondingAuthor":false,"prefix":"","firstName":"Goutam","middleName":"","lastName":"Mondal","suffix":""}],"badges":[],"createdAt":"2024-10-22 05:12:27","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5308483/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5308483/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10658-025-03001-8","type":"published","date":"2025-01-18T15:57:45+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":68799062,"identity":"10c92c27-4221-4309-a4d7-3169ff94d1ea","added_by":"auto","created_at":"2024-11-12 07:04:54","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":9434864,"visible":true,"origin":"","legend":"\u003cp\u003eMorphology of \u003cem\u003eTrichoderma\u003c/em\u003e isolates cultured on PDA incubated at 27±1°C for 7 days.\u003c/p\u003e\n\u003cp\u003e(a-b) Isolate T\u003csub\u003e1\u003c/sub\u003e. (c-d) Isolate T\u003csub\u003e2\u003c/sub\u003e. (e-f) Isolate T\u003csub\u003e3\u003c/sub\u003e. (g-h) Isolate T\u003csub\u003e4\u003c/sub\u003e. (i-j) Isolate T\u003csub\u003e5\u003c/sub\u003e. (k-l) Isolate T\u003csub\u003e6\u003c/sub\u003e. (m-n) Isolate T\u003csub\u003e7\u003c/sub\u003e. (o-p) Isolate T\u003csub\u003e8\u003c/sub\u003e. (q-r) Isolate T\u003csub\u003e9\u003c/sub\u003e. (s-t) Isolate T\u003csub\u003e10\u003c/sub\u003e (Scale bar = 20 µm)\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5308483/v1/9bcb7a22f3e942ec3352b433.png"},{"id":68799060,"identity":"8e664608-51bd-4692-aa6a-4ade4abcee57","added_by":"auto","created_at":"2024-11-12 07:04:54","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":7177069,"visible":true,"origin":"","legend":"\u003cp\u003eTest of efficacy of \u003cem\u003eTrichoderma\u003c/em\u003e isolates on \u003cem\u003eColletotrichum siamense \u003c/em\u003ein dual culture on PDA at 27±1°C for 7 days\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5308483/v1/fbfbd20758a86d75211f2170.png"},{"id":68799059,"identity":"09e77b98-3aaf-46a8-98aa-b0671ae07f6a","added_by":"auto","created_at":"2024-11-12 07:04:54","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":77501,"visible":true,"origin":"","legend":"\u003cp\u003eSporulation efficiency of \u003cem\u003eTrichoderma\u003c/em\u003eisolates on PDA assessed after 7 days of incubation at 27±1°C\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5308483/v1/c30ddd9185660d288b46b747.png"},{"id":68799063,"identity":"3ed6c73e-3f81-43c4-94ac-04a99a41227a","added_by":"auto","created_at":"2024-11-12 07:04:54","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":13158453,"visible":true,"origin":"","legend":"\u003cp\u003e(a-e) Application of \u003cem\u003eTrichoderma\u003c/em\u003eisolates on healthy Aloe vera plants by seedling root dip method under pot culture experiment. (f-i) Effects on plant growth parameters and disease severity after 120 days\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5308483/v1/9766ec40d2481176b98429e6.png"},{"id":68799061,"identity":"02d5e53a-b388-49f6-9e70-3044dbde778b","added_by":"auto","created_at":"2024-11-12 07:04:54","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":8384343,"visible":true,"origin":"","legend":"\u003cp\u003eMorphological characteristics of \u003cem\u003eTrichoderma\u003c/em\u003e \u003cem\u003easperellum\u003c/em\u003e on PDA incubated at 27±1°C for 7 days. a Front view. b Reverse view. c Colony under stereomicroscope. d Conidiophores and phialides. e Conidial mass. f Conidia\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5308483/v1/3b11bd9df121aeee39afa4f9.png"},{"id":68799065,"identity":"c782c688-8cdc-41cc-87ce-4fbf2a63bf86","added_by":"auto","created_at":"2024-11-12 07:04:54","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1835317,"visible":true,"origin":"","legend":"\u003cp\u003eConstruction of a phylogenetic tree using the internal transcribed spacer (ITS) region sequences from 30 nucleotide sequences, employing the maximum likelihood method in MEGA software. The analysis utilized the Kimura 2-parameter model with a gamma distribution and complete deletion of gaps, along with 1000 bootstrap replicates to assess reliability. \u003cem\u003eTrichoderma asperellum\u003c/em\u003e Isolate Kalyani-1 was highlighted in bold italic with its branch indicated in red, while \u003cem\u003eFusarium oxysporum\u003c/em\u003estrain ML-5-2 served as out group to root the tree. The scale bar indicates the number of substitutions at each position\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-5308483/v1/59a112496f0c836ff58ec95e.png"},{"id":74285057,"identity":"e57a3c86-4d7f-451d-a969-7902c6d33d9c","added_by":"auto","created_at":"2025-01-20 16:13:52","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":69596265,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5308483/v1/28e64aea-2aed-4429-bd1a-f9113022f7ff.pdf"}],"financialInterests":"","formattedTitle":"Evaluating bioactivity of Trichoderma asperellum against Colletotrichum siamense and its growth-promoting effects on Aloe vera (Aloe barbadensis Mill.)","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eTrichoderma\u003c/em\u003e is predominantly soil-dwelling, cosmopolitan, asexual filamentous fungi (with teleomorphic forms known as \u003cem\u003eHypocrea\u003c/em\u003e) that commonly inhabit decaying wood and other decomposing plant materials (Howell, 2003). These fungi are noted for their fast growth, featuring vibrant green conidia and conidiophores with a repetitively branched structure. \u003cem\u003eTrichoderma\u003c/em\u003e belongs to the domain Eukaryota and is part of the kingdom Fungi, and categorised within the division Ascomycota, subdivision Pezizomycotina, class Sordariomycetes, order Hypocreales, and family Hypocreaceae. \u003cem\u003eHypocrea/Trichoderma\u003c/em\u003e encompasses over 300 species identified both molecularly and morphologically, with many still awaiting formal description (Bissett et al., 2015). \u003cem\u003eTrichoderma\u003c/em\u003e employs multiple mechanisms to control phytopathogenic organisms such as competing with plant pathogens for resources, parasitizing and killing other fungi, producing antimicrobial substances, generating lytic enzymes, releasing secondary metabolites as well as induction of plant resistance (Vinale et al., 2008; Saravanakumar et al., 2016; Moosavi \u0026amp; Zare, 2020). \u003cem\u003eTrichoderma\u003c/em\u003e strains are effective bio-pesticides and bio-fertilizers due to their direct competition with pathogens (Ghisalberti \u0026amp; Sivasithaparam, 1991). Additionally, applying \u003cem\u003eTrichoderma\u003c/em\u003e strains to the soil and plants has been found to enhance both the productivity and quality of a variety of crops like cucumbers, tomatoes, carrots, beans, corn, cotton, tobacco, millet, and ornamental grasses which is likely due to the interaction between growth hormones produced by the fungi and the defence hormones triggered in the plant (Grant \u0026amp; Jones, 2009). It is recognized as an opportunistic plant symbiont and a potent mycoparasite. \u003cem\u003eTrichoderma\u003c/em\u003e species compete with other pathogens for nutrients, growth factors, and space and also produce highly effective siderophores that sequester iron, thereby hindering the growth of other filamentous fungi (Benitez et al., 2004). Secondary metabolites produced by \u003cem\u003eTrichoderma\u003c/em\u003e can be classified as volatile antibiotics, water-soluble compounds, and peptaibols, which are linear oligopeptides (Ghisalberti \u0026amp; Sivasithamparam, 1991).\u003c/p\u003e \u003cp\u003e \u003cem\u003eTrichoderma asperellum\u003c/em\u003e is widely regarded as a safe and effective biological control agent and does not persist in the food chain (Harman, 2006). These fungi can thrive in highly competitive environments and can counteract fungistatic effects (Benitez et al., 2004). Many strains exhibit resistance to a wide range of toxic substances, including metabolites produced by soil microorganisms or plants, as well as fungicides, herbicides, and antibiotics. \u003cem\u003eT. asperellum\u003c/em\u003e has been shown to stimulate cucumber plants to produce indole-3-acetic acid (IAA), gibberellins (GA), and abscisic acid (ABA), which contribute to growth promotion (Liu et al., 2022). In tomato seedlings treated with \u003cem\u003eT. asperellum\u003c/em\u003e, there were significant increases in plant height, stem diameter, soluble sugar content, and the absorption rate of available nitrogen. Additionally, the expression levels of hormone signalling-related genes, including JAR1, MYC2, NPR1, PR1, and GH3, were significantly elevated (Rawal et al., 2022). Another study indicated that \u003cem\u003eT. asperellum\u003c/em\u003e can enhance the expression of xylanase genes in poplar, demonstrating a notable growth-promoting effect (Karuppiah et al., 2019).\u003c/p\u003e \u003cp\u003eThe genus \u003cem\u003eColletotrichum\u003c/em\u003e includes several significant plant pathogenic fungi responsible for anthracnose and postharvest rots affecting a variety of fruits, vegetables, and ornamental plants, particularly in tropical and subtropical areas (Hyde et al., 2009). In fact, \u003cem\u003eColletotrichum\u003c/em\u003e species are recognized among the top 10 fungal pathogens of both scientific and economic importance (Dean et al., 2012). \u003cem\u003eColletotrichum siamense\u003c/em\u003e has been reported from countries like China, the USA, Africa, Vietnam, and Thailand and is known to cause diseases in a wide range of plants (Prihastuti et al., 2009) including Aloe vera. By deploying \u003cem\u003eTrichoderma\u003c/em\u003e, we can potentially mitigate the damage caused by \u003cem\u003eC. siamense\u003c/em\u003e on Aloe vera, offering a sustainable alternative to chemical treatments while simultaneously enhancing various plant growth parameters. In this study, we assessed the effectiveness of \u003cem\u003eTrichoderma asperellum\u003c/em\u003e against \u003cem\u003eC. siamense\u003c/em\u003e and its impact on various plant growth parameters in Aloe vera.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e \u003cb\u003eIsolation of\u003c/b\u003e \u003cb\u003eColletotrichum siamense\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003eColletotrichum siamense\u003c/em\u003e, the pathogen causing leaf spot in Aloe vera, was isolated in February 2022 from infected leaves at the Medicinal Plants Garden (22\u0026deg;59'21\"N latitude and 88\u0026deg;27'18\"E longitude) under the All India Coordinated Research Project on Medicinal and Aromatic Plants and Betel Vine, Directorate of Research, Bidhan Chandra Krishi Viswavidyalaya, Nadia, West Bengal, India. The pathogen is present year-round, peaking in winter. To isolate it, infected leaf pieces underwent surface sterilization with 0.1% mercuric chloride, followed by rinsing with sterile water, and cultured on Potato Dextrose Agar (PDA) with streptomycin sulphate. After 4\u0026ndash;5 days of incubation at 27\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C, single hyphal tips were sub-cultured to establish a pure culture and stored at 4\u0026deg;C. Identification of \u003cem\u003eC. siamense\u003c/em\u003e was confirmed through pathogenicity tests and molecular analysis of ITS, β-tubulin, and glyceraldehyde-3-phosphate dehydrogenase regions, with accession numbers OQ423208, PQ285628, and PQ285629 respectively.\u003c/p\u003e \u003cp\u003e \u003cb\u003eIsolation of\u003c/b\u003e \u003cb\u003eTrichoderma spp.\u003c/b\u003e\u003c/p\u003e \u003cp\u003eDuring February 2022, soil samples were randomly collected from a depth of 5 to 15 cm around the rhizosphere of Aloe vera plants at the Medicinal Plants Garden (22\u0026deg;59'21\"N latitude and 88\u0026deg;27'18\"E longitude) under the All India Coordinated Research Project on Medicinal and Aromatic Plants and Betel Vine, Directorate of Research, Bidhan Chandra Krishi Viswavidyalaya, Nadia, West Bengal, India. \u003cem\u003eTrichoderma\u003c/em\u003e species were isolated using serial dilution technique. For this purpose, \u003cem\u003eTrichoderma\u003c/em\u003e-specific media was prepared using the following composition: MgSO₄\u0026middot;7H₂O (0.2 g), K₂HPO₄ (0.9 g), KCl (0.15 g), NH₄NO₃ (3.0 g), Glucose (3.0 g), Agar (15 g), Rose Bengal (0.15 g), Chloramphenicol (0.25 g), and distilled water to a final volume of 1000 ml, adjusted to a pH of 6.5 (Elad et al., 1981).\u003c/p\u003e \u003cp\u003eThe initial dilution was created by adding 1 g of the representative soil sample to 9 ml of sterilized water in the first test tube, mixed thoroughly, and labelled as 10\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. Following this, 1 ml of the suspension from the first dilution was transferred to a second tube containing 9 ml of sterilized water, resulting in the 10\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003e dilution. This process was repeated until the original sample was diluted to 10\u003csup\u003e\u0026minus;\u0026thinsp;6\u003c/sup\u003e. From this 10\u003csup\u003e\u0026minus;\u0026thinsp;6\u003c/sup\u003e dilution, 1 ml was spread onto Petri dishes containing the \u003cem\u003eTrichoderma\u003c/em\u003e-specific media, supplemented with streptomycin sulphate (0.01%) as an antibiotic. The inoculated plates were then incubated in a BOD incubator at 27\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C for 4\u0026ndash;5 days to monitor fungal colony growth. After the incubation period, individual colonies exhibiting yellowish-green and whitish-green pigmentation were identified and transferred to sterilized Potato Dextrose Agar plates using a sterile loop to obtain pure cultures, which were subsequently stored in a refrigerator at 4\u0026deg;C for further analysis (Arumugam et al., 2013).\u003c/p\u003e \u003cp\u003eIdentification of Trichoderma isolates was based on cultural and microscopic characteristics. Colony morphology was assessed on PDA, and the lactophenol cotton blue staining technique was used for microscopic examination of conidiophores, conidia, and phialides, facilitating accurate identification of the isolates.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eDual culture technique\u003c/h2\u003e \u003cp\u003eThe antagonistic potential of the isolated \u003cem\u003eTrichoderma\u003c/em\u003e spp. was assessed against \u003cem\u003eC. siamense\u003c/em\u003e using dual culture technique (Ruocco et al., 2009). In this method, a 5 mm mycelial disc from 7 days old cultures of both the pathogen and the biocontrol agents was placed at opposite peripheries of 9 cm Petri plates containing PDA and the dual culture plates were incubated at 27\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C. Plates inoculated with only the disc of test pathogen served as control. There were three replications for each treatment. Antagonistic activity was evaluated by measuring the radial growth of the pathogen and comparing it with the growth observed in the control plates. Measurements of colony diameter for both the pathogen and the biocontrol agents were recorded after 3 and 7 days of incubation allowing for a comprehensive assessment of the interaction between the biocontrol agent and the pathogen. The percentage inhibition of radial growth (PIRG) of the pathogen was estimated by the formula given below (Vincent, 1947)\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003ePIRG = (C - T / C) × 100\u003c/h3\u003e\n\u003cp\u003eWhere, PIRG\u0026thinsp;=\u0026thinsp;Percentage inhibition of radial growth, C\u0026thinsp;=\u0026thinsp;Radial growth of pathogen in control and T\u0026thinsp;=\u0026thinsp;Radial growth of pathogen in presence of antagonist.\u003c/p\u003e \u003cp\u003eThe degree of antagonism between the biocontrol agent and test pathogen in dual culture (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) was scored on a scale of 1\u0026ndash;5 (Bell et al., 1982).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eBell\u0026rsquo;s scale for estimating the degree of antagonism in dual culture test\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eScale\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDegree of antagonisms\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1 =\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eThe antagonist fully outgrew the pathogen, covering the entire surface of the medium.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2 =\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eThe antagonist covered at least two-thirds of the medium's surface.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3 =\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBoth the antagonist and the pathogen occupied roughly half of the medium's surface (more than one-third but less than two-thirds), with neither organism showing dominance over the other.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4 =\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eThe pathogen occupied at least two-thirds of the medium's surface and seemed to resist encroachment.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5 =\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eThe pathogen fully outgrew the antagonist, covering the entire surface of the medium.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eAssessing sporulation capacity of\u003c/b\u003e \u003cb\u003eTrichoderma\u003c/b\u003e \u003cb\u003evia haemocytometer for enhancing Aloe vera growth in Pot Culture\u003c/b\u003e\u003c/p\u003e \u003cp\u003eSelecting \u003cem\u003eTrichoderma\u003c/em\u003e isolates based on their sporulation capacity is essential for enhancing the growth and health of Aloe vera in pot culture, as this influences the development of effective bioformulations (Jeyarajan \u0026amp; Nakkeeran, 2000). Isolates are cultured on media like Potato Dextrose Agar (PDA) for 7 days, with sporulation assessed through visual inspection and spore quantification using a haemocytometer. Spores are collected by adding sterile distilled water to the cultures and agitating gently. If the concentration is high, serial dilutions may be performed to ensure an appropriate count. A small volume of the spore suspension (typically 10 \u0026micro;L) is loaded onto the haemocytometer, covered with a coverslip, and observed under a light microscope where spore concentration (spores/mL) is determined using a formula that accounts for the total number of spores counted, the dilution factor, and the volume counted (Kamaruzzaman et al., 2016). High sporulation rates are prioritized, as these isolates are more likely to establish robust populations and effectively colonize the root zone of Aloe vera. The most promising isolates are then applied in pot culture experiments to evaluate their effects on plant growth, disease resistance, and overall health.\u003c/p\u003e \u003cp\u003e \u003cb\u003eEvaluation of\u003c/b\u003e \u003cb\u003eTrichoderma\u003c/b\u003e \u003cb\u003eIsolates for efficacy on Aloe vera\u003c/b\u003e \u003cb\u003ein-vivo\u003c/b\u003e\u003c/p\u003e \u003cp\u003eSelected \u003cem\u003eTrichoderma\u003c/em\u003e isolates were assessed for their effectiveness on Aloe vera plants grown in pots within a shade net house. The potting medium consisted of garden soil mixed with farmyard manure (FYM) in a 50:50 v/v ratio, which was used to cultivate young Aloe vera plants at 4\u0026ndash;5 leaf stage. Each \u003cem\u003eTrichoderma\u003c/em\u003e isolate was cultured in potato dextrose broth at 27\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C for 7 days. A talc-based bioformulation containing approximately 1\u0026times;10\u003csup\u003e8\u003c/sup\u003e spores per gram was prepared using spores harvested from the mycelial mat of the broth culture (Boblina et al., 2020). Each bioformulation was incorporated into the potting soil at a rate of 5 grams per kilogram (Kumar et al., 2014). Subsequently, Aloe vera seedlings were transplanted into the pots after soaking their roots in a solution mixed with the bioformulation @ 5 grams per litre of water. Each treatment included four replicates, while a control group was maintained separately. Plants were watered as necessary to ensure optimal moisture levels. Over the course of 120 days, the plants were monitored, and data were collected on disease symptoms, plant height, shoot and root biomass, and the number of leaves.\u003c/p\u003e \u003cp\u003e \u003cb\u003eIdentification of\u003c/b\u003e \u003cb\u003eTrichoderma asperellum\u003c/b\u003e\u003c/p\u003e \u003cp\u003eIdentification of \u003cem\u003eTrichoderma asperellum\u003c/em\u003e as an effective bio-control agent against \u003cem\u003eColletotrichum siamense\u003c/em\u003e and a growth promoter for Aloe vera was achieved through a combination of morphological assessments and molecular techniques. Key morphological characteristics, including colony colour, texture, and the structure of conidiophores, conidia, and phialides, were examined under stereo microscopes [Nikon (Model- SMZ 25)] and light microscopes [Nikon Trinocular Research Microscope (Model: Ci-L)]. To confirm the molecular identity, genomic DNA was extracted using the CTAB method (Doyle \u0026amp; Doyle, 1987). The amplification of the Internal Transcribed Spacer (ITS) region was conducted with the ITS1 and ITS4 primers. (White et al., 1990). Polymerase Chain Reactions (PCR) were conducted in a 25 \u0026micro;L reaction volume using the Veriti\u0026trade; PCR system. Each reaction included 2 \u0026micro;L of DNA sample, 1 \u0026micro;L each of forward and reverse primers, 12.5 \u0026micro;L of 2x Taq PCR Mix, and 8.5 \u0026micro;L of PCR grade water (GCC Biotech, India). The PCR protocol involved an initial denaturation at 95\u0026deg;C for 5 minutes, followed by 35 cycles of denaturation at 95\u0026deg;C for 1 minute, annealing at 60\u0026deg;C for 1 minute, and extension at 72\u0026deg;C for 1.5 minutes, concluding with a final elongation at 72\u0026deg;C for 8 minutes (White et al., 1990). The amplified PCR products were examined through electrophoresis on a 1.5% agarose gel, using a 100 bp DNA ladder for size determination (GCC Biotech, India). The PCR products were purified and sequenced using the Sanger method, with the resulting sequences compared to the NCBI GenBank database (Altschul et al., 1990) to confirm species identification and assess phylogenetic relationships. Further evolutionary analysis using MEGA X (Kumar et al., 2018) supported the identification of \u003cem\u003eT. asperellum\u003c/em\u003e.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eIsolation of\u003c/strong\u003e \u003cstrong\u003eTrichoderma\u003c/strong\u003e \u003cstrong\u003eisolates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA total of ten \u003cem\u003eTrichoderma\u003c/em\u003e species were isolated from the rhizosphere of Aloe vera using \u003cem\u003eTrichoderma\u003c/em\u003e selective medium (TSM) and subsequently subcultured on Potato Dextrose Agar (PDA). The isolates were designated as T\u003csub\u003e1\u003c/sub\u003e, T\u003csub\u003e2\u003c/sub\u003e, T\u003csub\u003e3\u003c/sub\u003e, T\u003csub\u003e4\u003c/sub\u003e, T\u003csub\u003e5\u003c/sub\u003e, T\u003csub\u003e6\u003c/sub\u003e, T\u003csub\u003e7\u003c/sub\u003e, T\u003csub\u003e8\u003c/sub\u003e, T\u003csub\u003e9\u003c/sub\u003e, and T\u003csub\u003e10\u003c/sub\u003e based on their morphological characteristics (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). To assess their biocontrol potential, the ability of these \u003cem\u003eTrichoderma\u003c/em\u003e isolates to inhibit the mycelial growth of \u003cem\u003eColletotrichum siamense\u003c/em\u003e was evaluated through dual culture experiments on PDA medium.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffect of\u003c/strong\u003e \u003cstrong\u003eTrichoderma\u003c/strong\u003e \u003cstrong\u003especies on mycelial growth of\u003c/strong\u003e \u003cstrong\u003eColletotrichum siamense in vitro\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll ten \u003cem\u003eTrichoderma\u003c/em\u003e isolates demonstrated an antagonistic effect against \u003cem\u003eColletotrichum siamense\u003c/em\u003e, effectively inhibiting its mycelial growth. After three days of inoculation, isolate T\u003csub\u003e3\u003c/sub\u003e exhibited the strongest antagonism, achieving a maximum reduction in colony growth to 2.03 cm, which corresponds to a 61.85% reduction. This was closely followed by isolate T\u003csub\u003e2\u003c/sub\u003e, which reduced the growth to 2.13 cm (59.97%), with both isolates showing statistically similar effects. The least inhibitory effect was noted in isolate T\u003csub\u003e5\u003c/sub\u003e, which achieved a 54.97% reduction. By the seventh day post-inoculation, isolates T\u003csub\u003e3\u003c/sub\u003e and T\u003csub\u003e7\u003c/sub\u003e recorded the highest reduction at 68.05%, with isolate T\u003csub\u003e4\u003c/sub\u003e at 67.63%, and isolates T\u003csub\u003e1\u003c/sub\u003e, T\u003csub\u003e2\u003c/sub\u003e, and T\u003csub\u003e10\u003c/sub\u003e each at 67.21%, all of which were statistically comparable. Isolate T\u003csub\u003e6\u003c/sub\u003e showed the lowest inhibition percentage at 60.07% (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e; Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). Additionally, the presence of the \u003cem\u003eTrichoderma\u003c/em\u003e isolates resulted in observable changes in the morphological features of the pathogen (Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eTest of efficacy of different \u003cem\u003eTrichoderma\u003c/em\u003e isolates against \u003cem\u003eColletotrichum siamense\u003c/em\u003e in dual culture test on PDA incubated at 27\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"7\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003eRadial Growth of Pathogen (cm)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIsolates\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3rd Day*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eInhibition\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7th Day*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eInhibition\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBell\u0026rsquo;s Scale\u003c/p\u003e\n \u003cp\u003e(on 7th day)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSporulation (x10\u003csup\u003e6\u003c/sup\u003e spores/ sq.cm)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e58.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e67.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.20\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e59.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e67.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e61.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e68.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.80\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e58.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e67.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e54.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e66.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.30\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e58.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e60.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.80\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e58.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e68.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.60\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e58.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e62.59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.50\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e58.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e65.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.20\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e10\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e58.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e67.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eS. Em\u0026plusmn;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC. D. (5%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC.V. (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.65\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003e*Average of three replications. Inhibition (%) was calculated over control\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eSome \u003cem\u003eTrichoderma\u003c/em\u003e isolates induced noticeable morphological changes in \u003cem\u003eColletotrichum siamense\u003c/em\u003e, particularly concerning colony colour and topography, compared to the control plate. The pure culture of the pathogen typically displayed a cottony, pink-white aerial mycelium with a flat topography. In contrast, isolate T\u003csub\u003e2\u003c/sub\u003e produced a cottony mycelium that was yellow to yellow-brown with a fluffy, raised appearance. Both isolates T\u003csub\u003e7\u003c/sub\u003e and T\u003csub\u003e9\u003c/sub\u003e exhibited yellow to brownish cottony mycelium with a similarly fluffy, elevated topography. Isolate T\u003csub\u003e10\u003c/sub\u003e, on the other hand, formed a cottony, dirty white mycelium, characterized by yellow margins and a fluffy, raised structure. These morphological alterations highlight the impact of \u003cem\u003eTrichoderma\u003c/em\u003e isolates on the pathogen\u0026apos;s growth characteristics (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eInfluence of \u003cem\u003eTrichoderma\u003c/em\u003e isolates on the growth behaviour of \u003cem\u003eColletotrichum siamense\u003c/em\u003e evaluated in dual culture on PDA, incubated at 27\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C for 7 days.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eTrichoderma\u003c/em\u003e isolates\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eColony Colour\u003c/p\u003e\n \u003cp\u003eFront plate\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eColony Colour\u003c/p\u003e\n \u003cp\u003eReverse plate\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTopography\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, white to off-white mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, white to off-white mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlat\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, yellow to yellow-brown mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, yellow to yellow-brown mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFluffy, raised\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAerial, white mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAerial, white mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlat\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, aerial, white mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, aerial, white mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlat\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOff-white to grey aerial mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOff-white to grey aerial mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFluffy, raised\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, pink-white, aerial mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, pink-white, aerial mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlat\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYellow to brownish cottony mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYellow to brownish cottony mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFluffy, raised\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePink-white cottony aerial mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePink-white cottony aerial mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlat\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, yellow to brownish mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, yellow to brownish mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFluffy, raised\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e10\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, dirty white to yellowish colour at the margin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, dirty white to yellowish colour at the margin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFluffy, raised\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, Pink-white aerial mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCottony, Pink-white aerial mycelium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlat\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe results from the \u003cem\u003ein vitro\u003c/em\u003e dual culture test indicated that \u003cem\u003eTrichoderma\u003c/em\u003e isolate T\u003csub\u003e3\u003c/sub\u003e exhibited the highest inhibition of colony growth of \u003cem\u003eColletotrichum siamense\u003c/em\u003e, achieving a reduction of 61.85%. This finding underscores the potential of isolate T\u003csub\u003e3\u003c/sub\u003e as an effective biocontrol agent against this pathogen.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSpore producing capacity of\u003c/strong\u003e \u003cstrong\u003eTrichoderma\u003c/strong\u003e \u003cstrong\u003espp.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn this study, the spore production capacity of the ten \u003cem\u003eTrichoderma\u003c/em\u003e spp. after a 7-day incubation period on PDA at 27\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C was observed. It was found that isolate T\u003csub\u003e3\u003c/sub\u003e exhibited highest spore concentration, with 2.8x10\u003csup\u003e6\u003c/sup\u003e spores per square centimetre of Petri plate. This was followed closely by isolates T\u003csub\u003e7\u003c/sub\u003e, T\u003csub\u003e5\u003c/sub\u003e, T\u003csub\u003e1\u003c/sub\u003e, and T\u003csub\u003e4\u003c/sub\u003e, which produced 2.6x10\u003csup\u003e6\u003c/sup\u003e, 2.3x10\u003csup\u003e6\u003c/sup\u003e, 2.2x10\u003csup\u003e6\u003c/sup\u003e, and 2.1x10\u003csup\u003e6\u003c/sup\u003e spores per square centimetre, respectively. In contrast, isolate T\u003csub\u003e8\u003c/sub\u003e demonstrated the lowest spore concentration, with only 0.5x10\u003csup\u003e6\u003c/sup\u003e spores per square centimetre (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eIn dual culture tests, all ten \u003cem\u003eTrichoderma\u003c/em\u003e isolates demonstrated effectiveness in suppressing the growth of \u003cem\u003eColletotrichum siamense in vitro\u003c/em\u003e. Among these isolates, five- T\u003csub\u003e1\u003c/sub\u003e, T\u003csub\u003e3\u003c/sub\u003e, T\u003csub\u003e4\u003c/sub\u003e, T\u003csub\u003e5\u003c/sub\u003e, and T\u003csub\u003e7\u003c/sub\u003e exhibited significantly higher sporulation capacities, producing greater numbers of spores compared to the other five isolates. Due to their superior performance in sporulation, these five isolates were selected for further evaluation in pot tests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePerformance of selected\u003c/strong\u003e \u003cstrong\u003eTrichoderma\u003c/strong\u003e \u003cstrong\u003eisolates in reducing leaf spot disease and their effects on Aloe Vera\u003c/strong\u003e \u003cstrong\u003ein vivo\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter 120 days, \u003cem\u003eTrichoderma\u003c/em\u003e inoculation significantly enhanced all growth metrics compared to the control (un-inoculated plants). Additionally, the inoculated plants exhibited reduced disease severity, indicating the protective role of \u003cem\u003eTrichoderma\u003c/em\u003e against the target pathogen. Isolate T\u003csub\u003e3\u003c/sub\u003e exhibited the highest plant weight at 243.90 g, reflecting a 144.30% increase over control, while isolate T\u003csub\u003e4\u003c/sub\u003e showed only a 15.50% increase with 115.33 g. T\u003csub\u003e3\u003c/sub\u003e also produced the longest shoot length (32.70 cm, 42.40% increase) and the greatest shoot biomass (228.17 g, 144.40% increase). In terms of root growth, T\u003csub\u003e3\u003c/sub\u003e had the longest root length (25.09 cm, 200% increase) and highest root biomass (15.98 g, 146.20% increase). Additionally, T\u003csub\u003e3\u003c/sub\u003e showed a 20.80% increase in the number of leaves, while T\u003csub\u003e1\u003c/sub\u003e and T\u003csub\u003e5\u003c/sub\u003e had no significant effect. Overall, isolate T\u003csub\u003e3\u003c/sub\u003e demonstrated superior performance in enhancing growth parameters, indicating its potential as an effective inoculant for Aloe vera (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e; Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eImpact of \u003cem\u003eTrichoderma\u003c/em\u003e isolates on various growth parameters of Aloe vera after 120 days under pot culture experiment\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"13\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eTrichoderma i\u003c/em\u003esolates\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eNo. of leaves\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIncrease\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLength of Shoot (cm)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIncrease\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eBiomass of shoot (gm)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIncrease\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLength of root (cm)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIncrease\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eBiomass of root (gm)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIncrease\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePlant weight (gm)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIncrease\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e28.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e111.59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e49.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e120.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.50\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e32.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e42.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e228.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e144.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e200.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e146.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e243.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e144.30\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e26.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e108.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e23.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e115.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.50\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e29.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e163.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e74.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e82.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e42.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e172.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e72.50\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e31.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e38.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e212.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e128.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e97.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e111.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e226.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e127.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e93.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e99.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eS. Em\u0026plusmn;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC. D. (5%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC. V. (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"13\"\u003e*\u003csub\u003eData were calculated per plant with four replications, and the percentage increase was determined in comparison to the control\u003c/sub\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cbr\u003e\u003c/div\u003e\u0026nbsp;\u003ctable id=\"Tab5\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eEffect of \u003cem\u003eTrichoderma\u003c/em\u003e isolates on Percent Disease Index (PDI) and Disease Incidence (%) with corresponding decrease (%) after 120 Days in pot culture experiment\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"5\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cem\u003eTrichoderma\u003c/em\u003e isolates\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eLeaf spot caused by \u003cem\u003eColletotrichum siamense\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePDI\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDecrease\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIncidence\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDecrease\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.83\u003c/p\u003e\n \u003cp\u003e(11.28)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e73.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19.17\u003c/p\u003e\n \u003cp\u003e(25.95)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e73.10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.21\u003c/p\u003e\n \u003cp\u003e(10.32)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e77.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.07\u003c/p\u003e\n \u003cp\u003e(23.62)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e77.44\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.33\u003c/p\u003e\n \u003cp\u003e(10.52)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e76.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.67\u003c/p\u003e\n \u003cp\u003e(24.09)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e76.61\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.83\u003c/p\u003e\n \u003cp\u003e(11.28)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e73.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19.17\u003c/p\u003e\n \u003cp\u003e(25.95)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e73.10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.00\u003c/p\u003e\n \u003cp\u003e(11.54)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e71.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.00\u003c/p\u003e\n \u003cp\u003e(26.57)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e71.93\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.25\u003c/p\u003e\n \u003cp\u003e(22.16)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e71.25\u003c/p\u003e\n \u003cp\u003e(57.69)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eS. Em\u0026plusmn;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC. D. (5%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC. V. (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003csup\u003e*Data were calculated per plant with four replications. The percentage decrease was determined relative to the control, with values in parentheses indicating angular transformation (arc sin)\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eFor \u003cem\u003eColletotrichum siamense\u003c/em\u003e, isolate T\u003csub\u003e3\u003c/sub\u003e exhibited the highest reduction in both PDI and disease incidence, achieving a decrease of 77.44%. In contrast, isolate T\u003csub\u003e7\u003c/sub\u003e showed the least reduction at 71.93% (Table \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e). Overall, isolate T\u003csub\u003e3\u003c/sub\u003e had a beneficial effect on Aloe vera plants, significantly lowering the PDI and disease incidence for the target pathogen.\u003c/p\u003e\n\u003cp\u003eIn conclusion, the findings from both the \u003cem\u003ein vitro\u003c/em\u003e dual culture tests and the \u003cem\u003ein vivo\u003c/em\u003e pot tests indicate that \u003cem\u003eTrichoderma\u003c/em\u003e isolate T\u003csub\u003e3\u003c/sub\u003e effectively suppressed the growth of \u003cem\u003eColletotrichum siamense\u003c/em\u003e. Additionally, isolate T\u003csub\u003e3\u003c/sub\u003e demonstrated the highest capacity for sporulation and promoted plant growth while inducing resistance in Aloe vera plants.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of\u003c/strong\u003e \u003cstrong\u003eTrichoderma\u003c/strong\u003e \u003cstrong\u003eisolate T\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/sub\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eTrichoderma\u003c/em\u003e isolate T\u003csub\u003e3\u003c/sub\u003e completely covered the Potato Dextrose Agar (PDA) plate by the fourth day when incubated at 27\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C. Initially, the colonies appeared white, but the mycelium became pale green to dark green after a few days. The conidiophores were branched and produced in pustules, ending in either a single phialide or a cluster of two to three diverging phialides (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e). The conidia (N\u0026thinsp;=\u0026thinsp;50) were dark green, globose to sub-globose, measuring 3.30\u0026thinsp;\u0026plusmn;\u0026thinsp;1.20 \u0026micro;m in length (range: 2.50\u0026ndash;4.60 \u0026micro;m) and 3.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95 \u0026micro;m in breadth (range: 2.15\u0026ndash;3.90 \u0026micro;m).\u003c/p\u003e\n\u003cp\u003eThe internal transcribed spacer (ITS) region was amplified and sequenced to ensure accurate species identification. BLASTn searches conducted in the NCBI database indicated that the ITS sequence of the isolate exhibited a high degree of similarity to other \u003cem\u003eT. asperellum\u003c/em\u003e strains, with identity percentages ranging from 99.84\u0026ndash;100%. Following the submission of the nucleotide sequences to the NCBI GenBank database, the accession number assigned was PP565067. The phylogenetic relationship of the isolate was analyzed using the Jukes-Cantor model within the maximum likelihood method, implemented in MEGA X software (Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eNumerous studies have investigated the biological control of \u003cem\u003eColletotrichum\u003c/em\u003e species using various strains of \u003cem\u003eTrichoderma\u003c/em\u003e. In dual culture experiments, it was observed that hyphal inhibition commenced upon contact between the antagonist and the pathogen (Almeida et al., 2007). \u003cem\u003eTrichoderma\u003c/em\u003e spp. effectively suppressed the hyphal growth of \u003cem\u003eColletotrichum\u003c/em\u003e spp. through mechanisms such as hyphal coiling, the induction of hyphal abnormalities, reduction in acervuli production, and lysis of both hyphae and sclerotia (Malathi et al., 2002). For instance, \u003cem\u003eTrichoderma viride\u003c/em\u003e demonstrated significant effectiveness, achieving 70.42% inhibition of \u003cem\u003eColletotrichum gloeosporioides\u003c/em\u003e (Patil et al., 2009). \u003cem\u003eT. asperellum\u003c/em\u003e strain T8a led to 80% growth inhibition of \u003cem\u003eC. gloeosporioides\u003c/em\u003e and an 85.18% inhibition of \u003cem\u003eC. asianum\u003c/em\u003e (De los Santos-Villalobos et al., 2013). Additionally, \u003cem\u003eT. asperellum\u003c/em\u003e isolate 21 also exhibited effective results in dual culture against these pathogens (Sharma \u0026amp; Prasad, 2018). Similarly, \u003cem\u003eT. longibrachiatum\u003c/em\u003e and \u003cem\u003eT. harzianum\u003c/em\u003e showed inhibition rates of 55.82% and 53.26% against \u003cem\u003eC. gloeosporioides\u003c/em\u003e, respectively (Valenzuela et al., 2015). Notably, isolates of \u003cem\u003eT. harzianum\u003c/em\u003e were found to be particularly effective, reducing the growth of \u003cem\u003eC. gloeosporioides\u003c/em\u003e by 42% (Prabakar et al., 2008).\u003c/p\u003e \u003cp\u003eCertain strains of \u003cem\u003eTrichoderma\u003c/em\u003e are known for their ability to establish robust and long-lasting colonization on root surfaces by penetrating the root epidermis. This process enhances root growth and improves resistance to abiotic stresses through better mineral absorption (Harman, 2006). The observed improvements in various plant growth parameters in the present study align with findings from several researchers (Chang et al., 1986; Ahmad \u0026amp; Baker, 1987; Yaqub \u0026amp; Shahzad, 2011).\u003c/p\u003e \u003cp\u003eAdditionally, it has been reported that plants inoculated with \u003cem\u003eTrichoderma\u003c/em\u003e exhibit increased disease resistance in plants, attributed to higher production of defence-related enzymes, including peroxidase, phenylalanine ammonia-lyase, and β-1,3-glucanase. These inoculations also contribute to earlier emergence and enhanced plant vigour (Jogaiah et al., 2013). When \u003cem\u003eTrichoderma asperellum\u003c/em\u003e was applied to the aerial parts of strawberry plants infected with anthracnose caused by \u003cem\u003eColletotrichum\u003c/em\u003e, a significant reduction in both disease incidence and severity was observed compared to untreated plants under greenhouse conditions (Kaissoumi et al., 2022).\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u003c/strong\u003e This work was supported by the Indian Council of Agricultural Research - All India Coordinated Research Project (AICRP) on Medicinal and Aromatic Plants and Betel vine, Directorate of Research, Bidhan Chandra Krishi Viswavidyalaya, Kalyani-741235, West Bengal, India, and National Medicinal Plant Board, Ministry of AYUSH, Government of India.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval:\u0026nbsp;\u003c/strong\u003eEthical approval was not applicable for this study since it exclusively involved using plant and microflora samples, which typically do not require human or animal ethics review.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contribution:\u0026nbsp;\u003c/strong\u003eAll authors have contributed to the manuscript\u0026apos;s study, conception, and design. Material preparation, data collection, and analysis- Ankur Mukhopadhyay, Soumik Mukherjee, Subham Dutta, and Goutam Mondal. Conceptualization- Ankur Mukhopadhyay and Goutam Mondal; Methodology- Ankur Mukhopadhyay, Soumik Mukherjee, and Subham Dutta. Writing and original draft preparation- Ankur Mukhopadhyay; Formal analysis and investigation- Ankur Mukhopadhyay and Goutam Mondal. Review and editing- Soumik Mukherjee, Subham Dutta, and Sahely Kanthal; Supervision- Goutam Mondal.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest:\u0026nbsp;\u003c/strong\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFinancial interest:\u0026nbsp;\u003c/strong\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e: The sequence data generated for this study has been submitted to the NCBI GenBank database with accession number PP565067 for ITS.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAhmad, J. S., \u0026amp; Baker, R. (1987). Rhizosphere competence of \u003cem\u003eTrichoderma harzianum\u003c/em\u003e. \u003cem\u003ePhytopathology\u003c/em\u003e, \u003cem\u003e77\u003c/em\u003e(2), 182\u0026ndash;189. https://doi.org/10.1094/Phyto-77-182 \u003c/li\u003e\n\u003cli\u003eAlmeida, F. B. D. R., Cerqueira, F. M., Silva, R. D. N., Ulhoa, C. J., \u0026amp; Lima, A. L. (2007). Mycoparasitism studies of \u003cem\u003eTrichoderma harzianum\u003c/em\u003e strains against \u003cem\u003eRhizoctonia solani\u003c/em\u003e: evaluation of coiling and hydrolytic enzyme production. \u003cem\u003eBiotechnology letters\u003c/em\u003e, 29, 1189\u0026ndash;1193. https://doi.org/10.1007/s10529-007-9372-z \u003c/li\u003e\n\u003cli\u003eAltschul, S. F., Gish, W., Miller, W., Myers, E. W., \u0026amp; Lipman, D. J. (1990). Basic local alignment search tool. \u003cem\u003eJournal of molecular biology\u003c/em\u003e, 215(3), 403\u0026ndash;410. https://doi.org/10.1016/s0022-2836(05)80360-2\u003c/li\u003e\n\u003cli\u003eArumugam, K., Ramalingam, P., \u0026amp; Appu, M. (2013). Isolation of \u003cem\u003eTrichoderma viride\u003c/em\u003e and \u003cem\u003ePseudomonas fluorescens\u003c/em\u003e organism from soil and their treatment against rice pathogens. \u003cem\u003eJournal of Microbiology and Biotechnology Research\u003c/em\u003e, \u003cem\u003e3\u003c/em\u003e(6),77\u0026ndash;81. \u003c/li\u003e\n\u003cli\u003eAzad, R., Hamza, A., Khatun, M. M., Nasrin, S., Tanny, T., Das, K. C., \u0026amp; Alam, I. (2020). First report of \u003cem\u003eColletotrichum siamense\u003c/em\u003e causing leaf spot in Aloe vera in Bangladesh. \u003cem\u003ePlant Disease\u003c/em\u003e, \u003cem\u003e104\u003c/em\u003e(11), 3058. https://doi.org/10.1094/PDIS-04-20-0857-PDN \u003c/li\u003e\n\u003cli\u003eBell, D. K., Wells, H. D., \u0026amp; Markham, C. R. (1982). In vitro antagonism of \u003cem\u003eTrichoderma\u003c/em\u003e species against six fungal plant pathogens. \u003cem\u003ePhytopathology\u003c/em\u003e, \u003cem\u003e72\u003c/em\u003e, 379\u0026ndash;382. https://doi.org/10.1094/Phyto-72-379 \u003c/li\u003e\n\u003cli\u003eBen\u0026iacute;tez, T., Rinc\u0026oacute;n, A. M., Lim\u0026oacute;n, M. C., \u0026amp; Cod\u0026oacute;n, A. C. (2004). Biocontrol mechanisms of \u003cem\u003eTrichoderma\u003c/em\u003e strains. \u003cem\u003eInternational Microbiology\u003c/em\u003e, \u003cem\u003e7\u003c/em\u003e(4), 249\u0026ndash;260.\u003c/li\u003e\n\u003cli\u003eBissett, J., Gams, W., Jaklitsch, W., \u0026amp; Samuels, G. J. (2015). Accepted \u003cem\u003eTrichoderma\u003c/em\u003e names in the year 2015. \u003cem\u003eIMA fungus\u003c/em\u003e, \u003cem\u003e6\u003c/em\u003e, 263\u0026ndash;295. https://doi.org/10.5598/imafungus.2015.06.02.02 \u003c/li\u003e\n\u003cli\u003eBoblina, B., Beura, S. K., Panda, A. G., \u0026amp; Mishra, M. K. (2020). Evaluation and assessment of shelf life of liquid substrates and talc formulation for mass production of native \u003cem\u003eTrichoderma\u003c/em\u003e spp. \u003cem\u003eJournal of Pharmacognosy and Phytochemistry\u003c/em\u003e, 9(3), 911\u0026ndash;915.\u003c/li\u003e\n\u003cli\u003eChang, Y. C., Baker, R., Kleifeld, O., \u0026amp; Chet, I. (1986). Increased growth of plants in the presence of the biological control agent \u003cem\u003eTrichoderma harzianum\u003c/em\u003e. \u003cem\u003ePlant Disease\u003c/em\u003e, \u003cem\u003e70\u003c/em\u003e(2):145\u0026ndash;148. https://doi.org/10.1094/PD-70-145 \u003c/li\u003e\n\u003cli\u003eDe los Santos-Villalobos, S., Guzm\u0026aacute;n-Ortiz, D. A., G\u0026oacute;mez-Lim, M. A., D\u0026eacute;lano-Frier, J. P., de-Folter, S., S\u0026aacute;nchez-Garc\u0026iacute;a, P., \u0026amp; Pe\u0026ntilde;a-Cabriales, J. J. (2013). Potential use of \u003cem\u003eTrichoderma\u003c/em\u003e \u003cem\u003easperellum\u003c/em\u003e (Samuels, Liechfeldt et Nirenberg) T8a as a biological control agent against anthracnose in mango (\u003cem\u003eMangifera indica\u003c/em\u003e L.). \u003cem\u003eBiological control\u003c/em\u003e, \u003cem\u003e64\u003c/em\u003e(1), 37\u0026ndash;44. https://doi.org/10.1016/j.biocontrol.2012.10.006 \u003c/li\u003e\n\u003cli\u003eDean, R., Van Kan, J. A., Pretorius, Z. A., Hammond‐Kosack, K. E., Di Pietro, A., Spanu, P. D., \u0026amp; Foster, G. D. (2012). The Top 10 fungal pathogens in molecular plant pathology. \u003cem\u003eMolecular plant pathology\u003c/em\u003e, \u003cem\u003e13\u003c/em\u003e(4), 414\u0026ndash;430. https://doi.org/10.1111/j.1364-3703.2011.00783.x \u003c/li\u003e\n\u003cli\u003eDoyle, J. J., \u0026amp; Doyle, J. L. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. \u003cem\u003ePhytochemical bulletin\u003c/em\u003e, \u003cem\u003e19\u003c/em\u003e, 11\u0026ndash;15.\u003c/li\u003e\n\u003cli\u003eElad, Y., Chet, I., \u0026amp; Henis, Y. (1981). A selective medium for improving quantitative isolation of \u003cem\u003eTrichoderma\u003c/em\u003e spp. from soil. \u003cem\u003ePhytoparasitica\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e, 59\u0026ndash;67. https://doi.org/10.1007/BF03158330 \u003c/li\u003e\n\u003cli\u003eGhisalberti, E. L., \u0026amp; Sivasithamparam, K. (1991). Antifungal antibiotics produced by \u003cem\u003eTrichoderma\u003c/em\u003e spp. \u003cem\u003eSoil biology and Biochemistry\u003c/em\u003e, \u003cem\u003e23\u003c/em\u003e(11), 1011\u0026ndash;1020. https://doi.org/10.1016/0038-0717(91)90036-J \u003c/li\u003e\n\u003cli\u003eGrant, M. R., \u0026amp; Jones, J. D. (2009). Hormone (dis) harmony moulds plant health and disease. \u003cem\u003eScience\u003c/em\u003e, \u003cem\u003e324\u003c/em\u003e(5928), 750\u0026ndash;752. https://doi.org/10.1126/science.1173771 \u003c/li\u003e\n\u003cli\u003eHarman, G. E. (2006). Overview of Mechanisms and Uses of \u003cem\u003eTrichoderma\u003c/em\u003e spp. \u003cem\u003ePhytopathology\u003c/em\u003e, \u003cem\u003e96\u003c/em\u003e(2), 190\u0026ndash;194. https://doi.org/10.1094/PHYTO-96-0190 \u003c/li\u003e\n\u003cli\u003eHowell, C. R. (2003). Mechanisms employed by Trichoderma species in the biological control of plant diseases: the history and evolution of current concepts. \u003cem\u003ePlant disease\u003c/em\u003e, \u003cem\u003e87\u003c/em\u003e(1), 4\u0026ndash;10. https://doi.org/10.1094/PDIS.2003.87.1.4 \u003c/li\u003e\n\u003cli\u003eHyde, K. D., Cai, L., Cannon, P. F., Crouch, J. A., Crous, P. W., Damm, U., \u0026amp; Tan, Y. P. (2009). \u003cem\u003eColletotrichum\u003c/em\u003e- names in current use. \u003cem\u003eFungal Diversity\u003c/em\u003e, \u003cem\u003e39\u003c/em\u003e(1), 147\u0026ndash;182.\u003c/li\u003e\n\u003cli\u003eJeyarajan, R., \u0026amp; Nakkeeran, S. (2000). Exploitation of microorganisms and viruses as biocontrol agents for crop disease management. In \u003cem\u003eBiocontrol Potential and its Exploitation in Sustainable Agriculture: Crop Diseases, Weeds, and Nematodes\u003c/em\u003e (pp. 95-116). Boston, MA: Springer US. https://doi.org/10.1007/978-1-4615-4209-4_8\u003c/li\u003e\n\u003cli\u003eJogaiah, S., Abdelrahman, M., Tran, L. S. P., \u0026amp; Shin-Ichi, I. (2013). Characterization of rhizosphere fungi that mediate resistance in tomato against bacterial wilt disease. \u003cem\u003eJournal of experimental botany\u003c/em\u003e, \u003cem\u003e64\u003c/em\u003e(12), 3829\u0026ndash;3842. https://doi.org/10.1093/jxb/ert212 \u003c/li\u003e\n\u003cli\u003eKaissoumi, H., Berber, F., Mouden, N., Ouazzani Chahdi, A., Albatnan, A., Ouazzani Touhami, A., \u0026amp; Douira, A. (2022). Effect of \u003cem\u003eTrichoderma asperellum\u003c/em\u003e on the development of strawberry plants and biocontrol of anthracnose disease caused by \u003cem\u003eColletotrichum\u003c/em\u003e \u003cem\u003egloeosporioides\u003c/em\u003e. In \u003cem\u003eInternational Conference on Advanced Intelligent Systems for Sustainable Development\u003c/em\u003e (pp. 609-622). Cham: Springer Nature Switzerland. https://doi.org/10.1007/978-3-031-35248-5_55 \u003c/li\u003e\n\u003cli\u003eKamaruzzaman, M., Rahman, M. M., Islam, M. S., \u0026amp; Ahmad, M. U. (2016). Efficacy of four selective \u003cem\u003eTrichoderma\u003c/em\u003e isolates as plant growth promoters in two peanut varieties. \u003cem\u003eInternational Journal of Biological Research\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e(2), 152\u0026ndash;156. https://doi.org/10.14419/ijbr.v4i2.6468 \u003c/li\u003e\n\u003cli\u003eKaruppiah, V., Vallikkannu, M., Li, T., \u0026amp; Chen, J. (2019). Simultaneous and sequential based co-fermentations of \u003cem\u003eTrichoderma asperellum\u003c/em\u003e GDFS1009 and \u003cem\u003eBacillus amyloliquefaciens\u003c/em\u003e 1841: a strategy to enhance the gene expression and metabolites to improve the bio-control and plant growth promoting activity. \u003cem\u003eMicrobial Cell Factories\u003c/em\u003e, \u003cem\u003e18\u003c/em\u003e, 1\u0026ndash;16. https://doi.org/10.1186/s12934-019-1233-7\u003c/li\u003e\n\u003cli\u003eKimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. \u003cem\u003eJournal of molecular evolution\u003c/em\u003e, \u003cem\u003e16\u003c/em\u003e, 111\u0026ndash;120. https://doi.org/10.1007/BF01731581\u003c/li\u003e\n\u003cli\u003eKumar, S., Stecher, G., Li, M., Knyaz, C., \u0026amp; Tamura, K. (2018). MEGA X: molecular evolutionary genetics analysis across computing platforms. \u003cem\u003eMolecular biology and evolution\u003c/em\u003e, \u003cem\u003e35\u003c/em\u003e(6), 1547\u0026ndash;1549. https://doi.org/10.1093/molbev/msy096 \u003c/li\u003e\n\u003cli\u003eKumar, S., Thakur, M., \u0026amp; Rani, A. (2014). \u003cem\u003eTrichoderma\u003c/em\u003e: Mass production, formulation, quality control, delivery and its scope in commercialization in India for the management of plant diseases. \u003cem\u003eAfrican journal of agricultural research\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(53), 3838\u0026ndash;3852. https://doi.org/10.5897/AJAR2014. 9061 \u003c/li\u003e\n\u003cli\u003eLiu, Y., He, P., He, P., Munir, S., Ahmed, A., Wu, Y., \u0026amp; He, Y. (2022). Potential biocontrol efficiency of \u003cem\u003eTrichoderma\u003c/em\u003e species against oomycete pathogens. \u003cem\u003eFrontiers in Microbiology\u003c/em\u003e, \u003cem\u003e13\u003c/em\u003e, 974024. https://doi.org/10.3389/fmicb.2022.974024 \u003c/li\u003e\n\u003cli\u003eMalathi, P., Viswanathan, R., Padmanaban, P., Mohanraj, D., \u0026amp; Sunder, A. R. (2002). Compatibility of biocontrol agents with fungicides against red rot disease of sugarcane. \u003cem\u003eSugar Tech\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e, 131\u0026ndash;136. https://doi.org/10.1007/BF02942694 \u003c/li\u003e\n\u003cli\u003eMoosavi, M. R., \u0026amp; Zare, R. (2020). Fungi as biological control agents of plant-parasitic nematodes. \u003cem\u003ePlant defence: biological control\u003c/em\u003e, 333\u0026ndash;384. https://doi.org/10.1007/978-3-030-51034-3_14 \u003c/li\u003e\n\u003cli\u003ePatil, C. U., Zape, A. S., \u0026amp; Wathore, S. D. (2009). Efficacy of fungicides and bioagents against \u003cem\u003eColletotrichum gloeosporioides\u003c/em\u003e causing blight in \u003cem\u003ePiper longum\u003c/em\u003e. \u003cem\u003eInternational Journal of Plant Protection\u003c/em\u003e, \u003cem\u003e2\u003c/em\u003e(1), 63\u0026ndash;66. \u003c/li\u003e\n\u003cli\u003ePrabakar, K., Raguchander, T., Saravanakumar, D., Muthulakshmi, P., Parthiban, V. K., \u0026amp; Prakasam, V. (2008). Management of postharvest disease of mango anthracnose incited by \u003cem\u003eColletotrichum gleosporioides\u003c/em\u003e. \u003cem\u003eArchives of Phytopathology and Plant protection\u003c/em\u003e, \u003cem\u003e41\u003c/em\u003e(5), 333\u0026ndash;339. https://doi.org/10.1080/03235400600793502\u003c/li\u003e\n\u003cli\u003ePrihastuti, H., Cai, L., Chen, H., McKenzie, E. H. C., \u0026amp; Hyde, K. D. (2009). Characterization of \u003cem\u003eColletotrichum\u003c/em\u003e species associated with coffee berries in northern Thailand. \u003cem\u003eFungal Diversity\u003c/em\u003e, \u003cem\u003e39\u003c/em\u003e(1), 89\u0026ndash;109.\u003c/li\u003e\n\u003cli\u003eRawal, R., Scheerens, J. C., Fenstemaker, S. M., Francis, D. M., Miller, S. A., \u0026amp; Benitez, M. S. (2022). Novel \u003cem\u003eTrichoderma\u003c/em\u003e isolates alleviate water deficit stress in susceptible tomato genotypes. \u003cem\u003eFrontiers in plant science\u003c/em\u003e, \u003cem\u003e13\u003c/em\u003e, 869090. https://doi.org/10.3389/fpls.2022.869090 \u003c/li\u003e\n\u003cli\u003eRuocco, M., Lanzuise, S., Vinale, F., Marra, R., Turr\u0026agrave;, D., Woo, S. L., \u0026amp; Lorito, M. (2009). Identification of a new biocontrol gene in \u003cem\u003eTrichoderma atroviride\u003c/em\u003e: the role of an ABC transporter membrane pump in the interaction with different plant-pathogenic fungi. \u003cem\u003eMolecular plant-microbe interactions\u003c/em\u003e, \u003cem\u003e22\u003c/em\u003e(3), 291\u0026ndash;301. https://doi.org/10.1094/MPMI-22-3-0291 \u003c/li\u003e\n\u003cli\u003eSaravanakumar, K., Yu, C., Dou, K., Wang, M., Li, Y., \u0026amp; Chen, J. (2016). Synergistic effect of \u003cem\u003eTrichoderma\u003c/em\u003e-derived antifungal metabolites and cell wall degrading enzymes on enhanced biocontrol of \u003cem\u003eFusarium oxysporum\u003c/em\u003e f. sp. \u003cem\u003ecucumerinum\u003c/em\u003e. \u003cem\u003eBiological control\u003c/em\u003e, \u003cem\u003e94\u003c/em\u003e, 37\u0026ndash;46. https://doi.org/10.1016/j.biocontrol.2015.12.001\u003c/li\u003e\n\u003cli\u003eSharma, K. S., \u0026amp; Prasad, L. (2018). Bioactivity of \u003cem\u003eTrichoderma asperellum\u003c/em\u003e against \u003cem\u003eColletotrichum asianum\u003c/em\u003e and \u003cem\u003eSclerotinia sclerotiorum\u003c/em\u003e. \u003cem\u003ePesticide Research Journal\u003c/em\u003e, \u003cem\u003e30\u003c/em\u003e(2), 251\u0026ndash;255. \u003cu\u003ehttps://doi.org/\u003c/u\u003e10.5958/2249-524X.2018.00040.7 \u003c/li\u003e\n\u003cli\u003eValenzuela, N. L., Angel, D. N., Ortiz, D. T., Rosas, R. A., Garc\u0026iacute;a, C. F. O., \u0026amp; Santos, M. O. (2015). Biological control of anthracnose by postharvest application of \u003cem\u003eTrichoderma\u003c/em\u003e spp. on maradol papaya fruit. \u003cem\u003eBiological Control\u003c/em\u003e, \u003cem\u003e91\u003c/em\u003e, 88\u0026ndash;93. https://doi.org/10.1016/j.biocontrol.2015.08.002 \u003c/li\u003e\n\u003cli\u003eVinale, F., Marra, R., Scala, F., Ghisalberti, E. L., Lorito, M., \u0026amp; Sivasithamparam, K. (2006). Major secondary metabolites produced by two commercial \u003cem\u003eTrichoderma\u003c/em\u003e strains active against different phytopathogens. \u003cem\u003eLetters in applied microbiology\u003c/em\u003e, \u003cem\u003e43\u003c/em\u003e(2), 143\u0026ndash;148. https://doi.org/10.1111/j.1472-765X.2006.01939.x \u003c/li\u003e\n\u003cli\u003eVincent, J. M. (1947). Distortion of fungal hyphae in the presence of certain inhibitors. \u003cem\u003eNature\u003c/em\u003e, \u003cem\u003e159\u003c/em\u003e(4051), 850\u0026ndash;850. https://doi.org/10.1038/159850b0 \u003c/li\u003e\n\u003cli\u003eWhite, T. J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR protocols a guide to methods and applications, 315\u0026ndash;322.\u003c/li\u003e\n\u003cli\u003eYaqub, F., \u0026amp; Shahzad, S. (2011). Efficacy and persistence of micobial antagonists against \u003cem\u003eSclerotium rolfsii\u003c/em\u003e under field conditions. \u003cem\u003ePakistan Journal of Botany\u003c/em\u003e, \u003cem\u003e43\u003c/em\u003e(5), 2627\u0026ndash;2634.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"european-journal-of-plant-pathology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ejpp","sideBox":"Learn more about [European Journal of Plant Pathology](http://link.springer.com/journal/10658)","snPcode":"10658","submissionUrl":"https://www.editorialmanager.com/ejpp/default2.aspx","title":"European Journal of Plant Pathology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Colletotrichum siamense, Trichoderma asperellum, serial dilution, dual culture, pot test, ITS","lastPublishedDoi":"10.21203/rs.3.rs-5308483/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5308483/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eLeaf spot disease caused by \u003cem\u003eColletotrichum siamense\u003c/em\u003e is a significant fungal threat to various plants, including Aloe vera. This study explores the biocontrol potential of \u003cem\u003eTrichoderma\u003c/em\u003e \u003cem\u003easperellum\u003c/em\u003e against \u003cem\u003eC. siamense\u003c/em\u003e while simultaneously evaluating the effects on Aloe vera growth parameters. Ten \u003cem\u003eTrichoderma\u003c/em\u003e isolates (T\u003csub\u003e1\u003c/sub\u003e to T\u003csub\u003e10\u003c/sub\u003e) were obtained from the rhizosphere of Aloe vera through serial dilution and assessed for their antagonistic activity using a dual culture technique. Among these isolates, five- T\u003csub\u003e1\u003c/sub\u003e, T\u003csub\u003e3\u003c/sub\u003e, T\u003csub\u003e4\u003c/sub\u003e, T\u003csub\u003e5\u003c/sub\u003e, and T\u003csub\u003e7\u003c/sub\u003e demonstrated the greatest suppression of radial growth of \u003cem\u003eC. siamense\u003c/em\u003e, along with high sporulation rates. In pot tests, isolate T\u003csub\u003e3\u003c/sub\u003e emerged as particularly effective, enhancing plant weight by 144.30%, shoot length by 42.40%, shoot biomass by 144.40%, root length by 200%, root biomass by 146.20%, and leaf number by 20.80%. Additionally, T\u003csub\u003e3\u003c/sub\u003e significantly reduced the severity of leaf spot disease, achieving a 77.44% decrease in disease severity. Morphological and molecular characterization confirmed isolate T\u003csub\u003e3\u003c/sub\u003e as \u003cem\u003eTrichoderma asperellum\u003c/em\u003e, with its internal transcribed spacer (ITS) sequence submitted to the NCBI GenBank and obtaining an accession number PP565067. These findings underscore the potential of \u003cem\u003eT. asperellum\u003c/em\u003e as an effective biocontrol agent, promoting healthier growth in Aloe vera while simultaneously managing leaf spot disease, making it a promising solution for sustainable agriculture practices.\u003c/p\u003e","manuscriptTitle":"Evaluating bioactivity of Trichoderma asperellum against Colletotrichum siamense and its growth-promoting effects on Aloe vera (Aloe barbadensis Mill.)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-12 07:04:49","doi":"10.21203/rs.3.rs-5308483/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision","date":"2024-12-02T06:08:08+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-10-31T13:43:41+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-10-31T11:37:13+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"European Journal of Plant Pathology","date":"2024-10-31T05:33:25+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-10-30T12:27:04+00:00","index":"","fulltext":""},{"type":"submitted","content":"European Journal of Plant Pathology","date":"2024-10-22T01:12:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"european-journal-of-plant-pathology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ejpp","sideBox":"Learn more about [European Journal of Plant Pathology](http://link.springer.com/journal/10658)","snPcode":"10658","submissionUrl":"https://www.editorialmanager.com/ejpp/default2.aspx","title":"European Journal of Plant Pathology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"02a81ad9-c91b-4a6d-810c-7b323b4006fe","owner":[],"postedDate":"November 12th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-01-20T16:06:57+00:00","versionOfRecord":{"articleIdentity":"rs-5308483","link":"https://doi.org/10.1007/s10658-025-03001-8","journal":{"identity":"european-journal-of-plant-pathology","isVorOnly":false,"title":"European Journal of Plant Pathology"},"publishedOn":"2025-01-18 15:57:45","publishedOnDateReadable":"January 18th, 2025"},"versionCreatedAt":"2024-11-12 07:04:49","video":"","vorDoi":"10.1007/s10658-025-03001-8","vorDoiUrl":"https://doi.org/10.1007/s10658-025-03001-8","workflowStages":[]},"version":"v1","identity":"rs-5308483","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5308483","identity":"rs-5308483","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}