Soil-Free Origins, Root-Deep Impact: Unlocking the Power of Native, Non-rhizosphere TrichodermaIsolates in Plant Growth and Yield under Agrochemical Environments

Soil-Free Origins, Root-Deep Impact: Unlocking the Power of Native, Non-rhizosphere TrichodermaIsolates in Plant Growth and Yield under Agrochemical Environments | 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 Article Soil-Free Origins, Root-Deep Impact: Unlocking the Power of Native, Non-rhizosphere Trichoderma Isolates in Plant Growth and Yield under Agrochemical Environments Ajith C R This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6653952/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This study investigates the compatibility of native non-rhizosphere Trichoderma isolates with agrochemicals and their impact on nutrient uptake and plant growth in tomato. Native isolates, particularly PSV and GMV, exhibited higher compatibility with agrochemicals compared to a commercial isolate, potentially due to their tolerance to chemical toxicity or the specific nature of the agrochemicals. A greenhouse experiment was conducted to assess the effects of three Trichoderma treatments—seed treatment (T1), combined seed and soil application (T2), and soil application (T3)—on tomato growth and nutrient content. T2 was the most effective treatment, leading to significant improvements in growth and nutrient uptake, particularly in iron (Fe), manganese (Mn), copper (Cu), and zinc (Zn), with PSV and SMV isolates being the most beneficial. Among these, PSV notably enhanced growth and fruit yield. The results highlight the potential of native non-rhizosphere Trichoderma spp. to promote nutrient uptake, improve plant biomass, and mitigate the adverse effects of soil-borne pathogens, suggesting their role as a sustainable biocontrol agent. Furthermore, the study emphasizes the importance of Integrated Disease Management (IDM) in reducing agrochemical reliance, with native Trichoderma isolates, especially PSV, identified as promising candidates for future field trials focused on both biocontrol and growth enhancement. Biological sciences/Microbiology Biological sciences/Plant sciences Earth and environmental sciences/Environmental sciences Trichoderma native non-rhizosphere compatibility micro-nutrient growth and yield Figures Figure 1 Figure 2 Figure 3 INTRODUCTION Natural agricultural soil hosts various beneficial microorganisms like Trichoderma , Pseudomonas , Bacillus spp., and arbuscular mycorrhizal fungi, which support plant health and soil fertility. These microbes promote plant growth by suppressing phytopathogens, enhancing nutrient uptake and cycling, producing growth hormones, and acting as biocontrol agents (Jeffries et al. 2003; Glic 1995). Among them, Trichoderma is a key biocontrol agent widely used in agriculture. It not only combats plant pathogens but also boosts plant growth (Shoresh et al. 2010), improves poor-quality seed vigour (Mastouri et al. 2010; Shoresh et al. 2010), enhances nitrogen-use efficiency (Shoresh et al . 2010; Harman 2011), and solubilizes micronutrients (Altomare et al. 1999). Trichoderma spp. is a fungal genus found globally in diverse environments like soil, forests, wood, and paper (Harman et al ., 2004). These fungi form symbiotic relationships with plants, encouraging robust root growth. Widely studied for their ability to enhance plant growth, Trichoderma species act as biofertilizers and biological control agents by producing antibiotics, parasitizing harmful fungi, and outcompeting plant pathogens (Adams et al., 2007; Bais et al., 2006). Some species promote growth by solubilizing phosphates and micronutrients like Fe, Mn, and Mg, improving nutrient uptake, and controlling root pathogens (Hoyos et al., 2009). They also enhance plant defense against biotic and abiotic stress (Mastouri et al., 2010). Recent applications focus on their role as biostimulants for seedling establishment, growth enhancement, and defense induction (Shanmugaiah et al ., 2009). While once attributed solely to indirect effects, it's now recognized that certain strains directly impact plant development and crop productivity (Harman, 2006). Trichoderma spp. enhance plant growth by solubilizing essential nutrients like Fe, Cu, Mn, Zn, and natural fertilizers such as rock phosphate and pyrite (Altomare et al ., 1999). Their nutrient-solubilizing ability offers a sustainable alternative to chemical fertilizers and pesticides. Since many soil nutrients are poorly soluble, Trichoderma aids in their availability by secreting organic acids that dissolve minerals and activate nutrients, improving soil nutrient cycling. Their strong colonization expands rhizosphere-soil interaction and boosts secretion of enzymes like sucrase, urease, and phosphatase, enhancing enzyme activity and nutrient uptake. Trichoderma also decomposes nitrogen compounds into plant-available forms while reducing NO₂ emissions (Jeffries et al ., 2003), and improves nutrient efficiency, lowering the need for nitrogen fertilizers (Harman, 2006). Inoculation with Trichoderma increases effective nutrient content and soil enzyme activity, supporting soil restoration and plant growth (Glic, 1995). Nutrients are vital for crop productivity and are classified as primary, secondary, and micronutrients based on crop requirements. Nutrient use efficiency refers to yield per unit of nutrient supplied from soil or fertilizer (Sayaji and Prasun, 2015). Common limiting nutrients include N, P, K, S, and micronutrients. Their availability is influenced by soil and rhizosphere microorganisms. Microbial enhancement of nutrient use efficiency is increasingly important due to declining soil fertility from intensive agriculture (Sayaji and Prasun, 2015). Previously, the same isolates were evaluated for compatibility with different fungicides (Ajith et al., 2024) and influence on major nutrient uptake and growth promotion in tomato (Ajith et al., 2023). In both study we found a significant compatibility and promotion of nutrient uptake and growth promotion. In this study, Trichoderma spp. were isolated from various local, non-rhizosphere sources and screened against soil-borne plant pathogens using the dual culture technique. Five effective local isolates were identified. To evaluate their impact on plant growth and nutrient use efficiency in tomato, a pot culture experiment was conducted under greenhouse conditions, as discussed below. MATERIALS AND METHOD Compatibility with agrochemicals Five effective non-rhizosphere Trichoderma isolates were cultured on sterile PDA medium for 3-4 days. T. viride , the commercial product, was used as a standard for comparison. Compatibility of the isolates with various agrochemicals (insecticides, bactericides, and herbicides) commonly used in tomato cultivation was tested using the poisoned food technique at concentrations of 50, 100, 200, 400, 600, 1000, and 1500 ppm. The chemicals were mixed with sterilized PDA medium, and mycelial discs of Trichoderma were inoculated into the plates. The plates were incubated at 28±2°C, and mycelial growth was measured to calculate percent inhibition. Per cent inhibition of Trichoderma was calculated by formula as mentioned below given by Vincent (1927). I = (C-T/C) × 100 Where, I = Per cent inhibition C = Radial mycelial growth of Trichoderma isolates in control T = Radial mycelial growth of Trichoderma isolates in treatments Influence on nutrient use efficiency in tomato Five Trichoderma isolates and one commercial formulation were evaluated for their effect on nutrient use efficiency (NUE) in tomato under greenhouse conditions using pot culture during the summer of 2020 at the College of Agriculture, V.C. Farm, Mandya. The tomato cultivar 'Abhinav' was used, with treatment combinations detailed in the accompanying table. Mass multiplication of Trichoderma and quality test All the Trichoderma isolates were mass multiplied on PDB media and formulated into talc formulation by mixing in 1:2 ratio (Trichoderma culture: talc powder). Formulation is maintained with 2*10 6 cfu/ml in all the isolates (Plate 2). Seed treatment Tomato seeds of cultivar Abhinav was treated with jiggery syrup and then with talc formulation @ 4g/kg seeds. Later seeds were shade dried and sown in pro trays to raise seedlings. Soil application The talc formulation was mixed with potting mixture at 10g/kg of soil 10days before transplanting seedlings. Soil and Plant analysis Soil sample was collected before Trichoderma application and after Trichoderma application, just before transplanting tomato seedlings. Tomato plant sample was collected 30 days after transplanting for nutrient analysis. Above ground plant sample was collected and fresh weight was taken using weighing balance. Then the sample was dried in hot air oven at 70±2⁰C for 48 hours. Then plant sample was ground to fine powder using grinding mill and stored for analysis. Soil analysis DTPA – Extractable micronutrients Micronutrient cations (Fe, Mn, Zn and Cu) from soil were extracted with DTPA extractant (0.005 M Diethylene Triamine Penta Acetic acid + 0.01 M CaCl 2 + 0.1 M Triethanolamine buffered to pH 7.3) at 1:2 soil to extractant ratio as described by Lindsay and Norvell (1978). The concentration of these cations was determined by atomic absorption spectrophotometer under suitable measuring conditions (Page et al., 1982). Digestion of plant samples with Di-acid mixture Powdered plant sample of 1.0 g were pre-digested with conc. HNO 3 overnight and then digested with di-acid mixture containing HNO 3 and HClO 4 in the proportion of 9:4 till a snow white residue was obtained (Piper, 1966). The volume of the digest was made to 100 ml with distilled water and used for total elemental analysis. Plant analysis for micronutrients (Fe, Mn, Zn and Cu) Micronutrients ( Fe, Mn, Zn and Cu) concentration in the di-acid extract was determined using Atomic Absorption Spectrophotometer (AAS) under suitable measuring conditions (Lindsay and Norwell, 1978; Page et al., 1982). Plant growth and yield Observations were taken on growth parameters of tomato like plant height, plant fresh and dry weight, root length, root fresh and dry weight. Tomato fruit was harvested regularly starting from 75 DAT and yield from different treatments was compared with control. Experimental results Compatibility with insecticides: Among seven insecticides (Table 1), chlorpyrifos 50% EC caused minimal mycelial inhibition (Plate 1) (<10%) up to 600 ppm in native Trichoderma isolates, with GMV showing the least inhibition (0–8.25%). The commercial isolate showed the highest inhibition (3–49.33%) across concentrations. Similarly, thiamethoxam 25% WG caused <5% inhibition up to 400 ppm, with PSV showing the least (20.35%) and the commercial isolate the highest (51.24%) inhibition at 1500 ppm. Flubendiamide 20% WG showed no mycelial inhibition up to 200 ppm in SMV, SDKd, GMV, and CPV isolates. At 1500 ppm, inhibition ranged from 16.38% to 68.36%, with GMV showing the least and PSV the highest inhibition. Fipronil 5% SC was compatible with all Trichoderma isolates, showing no inhibition up to 400 ppm in SDKd, GMV, and PSV. At 1500 ppm, these isolates showed low inhibition (13.65–33.85%), while CPV and the commercial isolate showed moderate inhibition (62.33–67.88%). Chlorantraniliprole 20% SC also showed compatibility, with GMV showing least inhibition (0–11.54%) and the commercial isolate the highest (21.85–67.63%). At 1500 ppm, SDKd and GMV remained highly compatible. Emamectin benzoate 5% SG showed good compatibility, with GMV, SDKd, and PSV displaying 0% inhibition up to 200 ppm and <40% at 1500 ppm. Other isolates, including the commercial one, showed moderate incompatibility (<70% inhibition). Imidacloprid 20% SC caused the least inhibition in GMV (0–50.44%) and the highest in the commercial isolate (0–58.33%). At 1500 ppm, it was moderately incompatible with all Trichoderma isolates. Compatibility with bactericide and herbicide: To assess compatibility, one bactericide and one herbicide were tested on native Trichoderma isolates at seven concentrations (Table 2). Both were compatible. The herbicide butachlor showed no mycelial inhibition at 50 and 100 ppm in all isolates, with GMV showing 0% inhibition up to 600 ppm. At 1500 ppm, GMV showed the least inhibition (13.52%) and the commercial isolate the highest (58.31%). For the bactericide K-cyclin, SDKd and GMV showed 0% mycelial inhibition up to 200 ppm, while PSV and SMV showed no inhibition up to 100 ppm. The bactericide remained compatible up to 1000 ppm in most isolates, except PSV, CPV, and the commercial isolate, which showed moderate inhibition (51.24–63.37%). At 1500 ppm, SDKd had the least inhibition (41.23%) and PSV the highest (68.32%), with the commercial isolate showing 67.65%. The compatibility of seven insecticides, one herbicide, and one bactericide with native Trichoderma isolates and a commercial isolate was tested. All agrochemicals were compatible with varying mycelial inhibition. Chlorpyrifos 50% EC was the most compatible, followed by thiamethoxam 25% WG and flubendiamide 20% WG. Imidacloprid 20% SC and emamectin benzoate 5% SG were the least compatible, showing moderate incompatibility at 1500 ppm. Praful and Mane (2017) observed that chlorantraniliprole 20% SC, emamectin benzoate 5% SG, and chlorpyrifos 50% EC were highly compatible with T. pseudokoningii at 10-2000 ppm. Similarly, chlorantraniliprole 20% SC and imidacloprid 20% SC were compatible with T. harzianum , with 85mm colony growth, while thiamethoxam 25% WG showed 3.82% mycelial inhibition (Thiruchchelvan et al., 2013). Madhusudhan et al . (2010) found that thiamethoxam 25% WG and fipronil 5% SC were compatible with T. viride , with 12% and 14.3% inhibition. Theertha et al. (2017) noted that chlorpyrifos, fipronil, and flubendiamide were compatible with T. asperellum with less than 15% inhibition up to 800 ppm.When compared among all the agrochemicals, SDKd, GMV and PSV were the isolates which showed compatibility with most number of agrochemicals, followed by SMV and CPV. However, the commercial isolate showed compatibility with least number of agrochemicals (Supplementary table 1 & 2). The isolate SDKd showed high compatibility with insecticides chlorpyrifos, emamectin benzoate, imidacloprid, thiamethoxam, flubendiamide, chlorantraniliprole, fipronil, and the herbicide butachlor, and moderate compatibility with bactericide K-cycline. It showed moderate incompatibility with tebuconazole and difenconazole. The GMV isolate was highly compatible with chlorpyrifos, flubendiamide, butachlor, and bactericide K-cycline. It was also compatible with emamectin benzoate, fipronil, thiamethoxam, and chlorantraniliprole, with moderate compatibility with imidacloprid. PSV showed high compatibility with chlorpyrifos, fipronil, and butachlor, with moderate compatibility with thiamethoxam, emamectin benzoate, and chlorantraniliprole, while moderate incompatibility was seen with flubendiamide and K-cycline. CPV exhibited high compatibility with chlorpyrifos and butachlor, and moderate compatibility with thiamethoxam, emamectin benzoate, chlorantraniliprole, and imidacloprid. SMV showed high compatibility with thiamethoxam and butachlor, and compatibility with chlorantraniliprole, fipronil, flubendiamide, imidacloprid, chlorpyrifos, emamectin benzoate, and K-cycline. The commercial isolate was compatible with chlorpyrifos, thiamethoxam, flubendiamide, emamectin benzoate, imidacloprid, chlorantraniliprole, and butachlor, with moderate compatibility with fipronil and K-cycline. Influence on nutrient use efficiency in tomato Germination percentage: Tomato seeds (variety Abhinav) treated with different native Trichoderma isolates were sown in pro-trays and monitored for germination. All Trichoderma treatments significantly improved germination compared to the control (T4). Isolates GMV and PSV showed the highest germination rates at 92.85% and 90%, respectively, followed by SMV (90%). SDKd had the lowest among native isolates (88.57%), while the commercial isolate recorded the lowest overall (85.71%). Nutrient status of soil before Trichoderma treatment Totally four micro nutrients viz., DTPA exchangeable Zn, Cu, Mn and Fe were assessed in the laboratory using standard protocols (Table 3). The soil contained 3.10 mgkg -1 of Zn, 1.39 mgkg -1 of copper (Cu), 5.86 mgkg -1 of manganese (Mn) and 9.13 mgkg -1 of iron (Fe). Nutrient status of soil after Trichoderma treatment: Rhizosphere soil analysis 30 days post-transplanting showed increased nutrient content after Trichoderma treatment compared to pre-treatment levels (Table 4). Zinc (Zn) (mgkg -1 of soil): Soil Zn content varied significantly across isolates, treatments, and their interactions. In T1, SDKd and commercial isolates showed the highest Zn (2.90 mgkg⁻¹), while CPV recorded the lowest (2.50 mgkg⁻¹). In T2, SMV and GMV had the highest Zn (2.70 mgkg⁻¹), and CPV the lowest (2.30 mgkg⁻¹). In T3, SMV showed the highest Zn (2.90 mgkg⁻¹), followed by commercial (2.70 mgkg⁻¹), while CPV and GMV had the lowest (2.40 mgkg⁻¹). Copper (Cu) (mgkg -1 of soil): Cu content varied across treatments and isolates. In T1, commercial and SMV showed highest Cu (1.35 and 1.33 mgkg⁻¹), while GMV and CPV had the lowest (1.25 and 1.27 mgkg⁻¹). In T2, commercial and SMV again led (1.33 and 1.30 mgkg⁻¹), with GMV, CPV, and PSV lowest (1.22–1.25 mgkg⁻¹). In T3, SMV had highest Cu (1.31 mgkg⁻¹), followed by commercial (1.30 mgkg⁻¹), while GMV and CPV showed lowest (1.23–1.24 mgkg⁻¹). Manganese (mgkg -1 of soil): Mn content differed among isolates and treatments. In T1, commercial and SDKd showed highest Mn (5.71 and 5.68 mgkg⁻¹), while GMV had the lowest (5.61 mgkg⁻¹). PSV and CPV recorded 5.62 mgkg⁻¹. In T2, commercial and SDKd again led (5.68 and 5.65 mgkg⁻¹), with GMV lowest (5.57 mgkg⁻¹). SMV, PSV, and CPV followed at 5.60 mgkg⁻¹. In T3, commercial (5.65 mgkg⁻¹) and SMV (5.63 mgkg⁻¹) had highest Mn, while SDKd, GMV, and CPV recorded the lowest (5.60 mgkg⁻¹). Iron (Fe) (mgkg -1 of soil): Fe content varied across treatments. In T1, commercial and CPV recorded highest Fe (9.01 and 8.97 mgkg⁻¹), while GMV had the lowest (8.85 mgkg⁻¹). In T2, commercial (9.03 mgkg⁻¹) and CPV (8.95 mgkg⁻¹) again showed higher Fe levels, with SMV and GMV the lowest (8.87 mgkg⁻¹). In T3, CPV and commercial both recorded 8.97 mgkg⁻¹, while SMV and GMV had the lowest Fe (8.81 mgkg⁻¹). No significant nutrient differences were noted across treatments and isolates, likely due to enhanced availability and uptake following Trichoderma application. Nutrient status of tomato plants 30 days after transplanting Tomato plant analysis showed significant nutrient differences between Trichoderma treatments and control. Four micronutrients viz., Zn, Cu, Mn, and Fe were examined and discussed (Table 5; Supplementary table 3). Zinc (Zn): Zn content in tomato tissues (mgkg⁻¹) was highest in T2, with GMV recording 30.94 and CPV the lowest at 22.81. The commercial isolate showed 22.90. In T1, GMV and CPV recorded 24.57 and 21.46, respectively, while the control showed the lowest Zn content at 17.86. Copper (Cu) T2 showed highest Cu content, with PSV and commercial isolates recording 18.60 and 22.90 mgkg⁻¹, respectively. In T1 and T3, PSV had the highest Cu (12.63 and 15.51 mgkg⁻¹), while the control recorded 10.25 mgkg⁻¹. Manganese (Mn) T2 showed the highest Mn content, with SMV and commercial isolates recording 84.23 and 79.56 mgkg⁻¹, respectively. In T1 and T3, the highest Mn was in SDKd (68.35 mgkg⁻¹) and SMV (80.62 mgkg⁻¹). The control showed 57.74 mgkg⁻¹. Iron (Fe): In T2, GMV and commercial isolates recorded the highest Fe content (172.36 and 169.62 mgkg⁻¹). In T1 and T3, GMV and PSV had the highest Fe levels (131.23 and 170.25 mgkg⁻¹), while the control recorded 121.36 mgkg⁻¹. Similar findings were reported by Rudresh et al. (2005), who observed that native isolates of Trichoderma spp. were capable of solubilizing insoluble tricalcium phosphate to varying degrees. In pot culture experiments using rock phosphate, enhanced phosphorus uptake was recorded in plants treated with T. harzianum (PDBCTH 10), followed by T. virens (PDBCTVs 12) and T. viride (TV 97). Rasool et al. (2011) found that tomato plants grown in soil fortified with Trichoderma spp. T and T. harzianum T969 exhibited increased uptake of nutrients such as nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg), with soil application proving more effective than seed treatment. Bombiti et al . (2011) reported that pre-sowing application of T. harzianum alone led to improved shoot uptake of N, P, Ca, Mg, and sulfur (S). Under glasshouse conditions, Inbar et al . (1994) demonstrated that Trichoderma significantly increased phosphorus and potassium content in tomato leaves even without fertilizer application. Ying et al. (2018) noted that the T. asperellum CHF 78 isolate, which produces enzymes such as cellulases, chitinases, proteases, and siderophores along with indole acetic acid (IAA), substantially enhanced the uptake of P, K, Mg, and Zn. Similarly, Singh et al . (2014) observed that a combination of T. harzianum isolates (BHU51 + BHU105) increased the mineral content of tomato plants—N, P, K, Ca, Mg, S, Zn, Cu, Mn, and Fe—by 40–50% compared to the untreated control. Tomato growth promotion by Trichoderma treatment Tomato growth parameters viz., plant height, fresh and dry weights of plant and root, root length, and fruit yield were recorded at intervals to assess how Trichoderma isolates affect growth and nutrient uptake from soil (Plate 3 and 4). Plant growth parameters: Tomato growth parameters such as plant height (cm) at 15, 30, 45, and 90 DAT and plant fresh/dry weight at 90 DAT showed significant differences among isolates and treatments (Supplementary table 4). The combination treatment (T2) was most effective. At 15 DAT, isolate GMV gave the highest plant height (43.22 cm), followed by PSV (42.68 cm) and CPV (42.53 cm), with GMV and PSV maintaining superior heights at 90 DAT (173.36 cm and 172.32 cm) compared to the control (30 cm at 15 DAT, 111.67 cm at 90 DAT). The commercial isolate performed poorest. In T1 and T3, isolate SMV was best at 90 DAT. Trichoderma notably enhanced plant height via improved nutrient uptake. At crop maturity, plant fresh and dry weight data (Supplementary table 5; Fig. 1) showed that the combined seed and soil treatment (T2) was most effective. Isolate GMV yielded the highest fresh (263.68g) and dry weight (53.22g), followed by PSV (227.69g and 43.68g). The commercial isolate showed lower values (195.66g and 37.68g), and the control was significantly inferior (142.66g and 22.75g). In T1 and T3, GMV again outperformed others (196.56g and 245.68g fresh weight; 29.56g and 40.88g dry weight), while the commercial isolate recorded lower values (144.69g and 187.58g fresh; 23.15g and 28.22g dry). Root growth parameters In order to take observations on root length, root fresh and dry weight, the plants were uprooted at maturity of the crop (90DAT) and observed for various parameters (Supplementary table 6; Fig. 2; Plate 4). With respect to all parameters, T2 was significantly superior over other treatments and control. In T2, SMV was the top-performing isolate with 30.45 cm root length, 46.68 g fresh weight, and 14.26 g dry weight, followed by PSV (30.11 cm, 42.12 g, and 12.41 g). The commercial isolate recorded lower values (26.24 cm, 26.77 g, and 7.29 g), while the control had the least (17.88 cm, 15.65 g, and 3.33 g). In T1, GMV had the longest roots (23.13 cm), while PSV showed the highest root fresh and dry weights (21.10 g and 4.54 g). In T3, SMV performed best (28.11 cm, 36.14 g, and 9.78 g). Tomato fruit yield Trichoderma-enhanced nutrient uptake led to improved tomato growth and yield. Yield analysis (Table) showed that both treatments and isolates had significant effects. The combination treatment (T2) was most effective, with GMV yielding 521.62 g per plant, followed by PSV (512.45 g), outperforming the commercial isolate (481.23 g), while the control had the lowest yield (318.27 g). In T1, GMV gave the highest yield (367.63 g), and in T3, GMV and PSV performed similarly (484.65 g and 484.38 g, respectively). Similar findings were reported by Rasool et al. (2011), who observed significant increases in shoot length and shoot/root fresh and dry weights in Trichoderma-fortified soils. Khoshmanzar et al. (2020) found that T. asperellum B1092 enhanced tomato shoot and root growth over the control. Treatments with T. longibrachiatum KH (T1), MA (T2), and T. harzianum (T3) increased shoot dry weight by 97.2%, 17.2%, and 18.96% over the negative control, and by 32.81%, 19.41%, and 20.42% over the positive control. Ozbay and Newman (2004) tested T. harzianum strains (Plant Shield, T22, T95) on 18-day-old tomato seedlings. After six weeks, seedling emergence was 100%, 83.33%, 88.89%, and 83% for Plant Shield, T22, T95, and control, respectively. T95 showed the highest plant height (34.35 cm) and the greatest shoot and root weight (43.10 g, 4.68 g, 9.13 g, and 0.87 g). Greenhouse results showed that T. pseudokoningii Rifai enhanced nutrient availability in soil (K, Ca, P, Zn, Fe) and in tomato plant tissue. Soil levels were 0.750 cmol/kg K, 10.05 cmol/kg Ca, 41.5 ppm P, 22 ppm Zn, and 12 ppm Fe. Tomato tissues contained 2.195% N, 0.046% P, 2.290% Ca, 0.0075% Zn, and 0.035% Fe. These nutrients promoted higher dry matter production (14g in control vs. 9g in Trichoderma treatment), resulting in earlier flowering and better fruit yield (Cuevas, 2006). Abul et al . (2012) found that treating tomato seeds with native Trichoderma isolates (T85, M14, M2_14) significantly increased plant height (M2_14: 25.22 cm, T85: 28.28 cm vs. 22.8 cm control) and root length (M14: 10.5 cm, T85: 10.85 cm vs. 10.05 cm control). T85 also enhanced root fresh/dry weight (4.45g/2.93g vs. 1.91g/0.91g control). Vinale et al. (2008) found that rhizosphere-competent Trichoderma isolates produce diffusible metabolites that act as plant growth regulators (auxins or auxin-like compounds), enhancing the growth of colonized plants. Correlating nutrient use efficiency with plant growth and yield The correlation analysis revealed significant relationships between soil and plant nutrient concentrations and various growth and yield parameters (Table 7; Fig. 3). Notably, plant zinc concentration showed a strong positive correlation with fruit yield (r = 0.919), plant dry weight (r = 0.938), and plant fresh weight (r = 0.878), suggesting that higher zinc accumulation in plant tissue is closely associated with increased biomass production and fruit output. Similarly, plant height exhibited strong positive correlations with root length (r = 0.927) and root dry weight (r = 0.827), indicating that taller plants tend to develop more extensive root systems. On the other hand, soil copper, manganese, and iron levels displayed negative correlations with most growth traits and yield. Specifically, soil copper and manganese had strong negative correlations with fruit yield (r = -0.752 for both), while soil iron showed a pronounced negative association with plant height (r = -0.865), implying that elevated levels of these micronutrients in soil might hinder plant development and productivity. These findings highlight the importance of maintaining optimal micronutrient balance in the soil to ensure healthy plant growth and maximize yield potential. Correlation analysis showed that plant zinc levels were strongly linked to dry weight (r = 0.938), fresh weight (r = 0.878), and fruit yield (r = 0.919), reinforcing its role in enhancing biomass and productivity (Sharma et al ., 2022). Plant height correlated positively with root length (r = 0.927) and root dry weight (r = 0.827), as noted by Li et al . (2021). In contrast, soil copper and manganese negatively affected fruit yield (r = -0.752) and plant height (r = -0.656), aligning with reports of phytotoxicity (Wang et al ., 2022; Zhang et al ., 2021). Soil iron also showed a strong negative correlation with plant height (r = -0.865), supporting findings by Kumar et al . (2023). These patterns highlight the critical role of balanced micronutrient availability in supporting optimal plant growth and yield. The clustered correlation heat map highlights key relationships between soil nutrients, plant traits, and yield. Strong positive correlations, such as between Plant Zn and both Plant dry weight (0.938) and Fruit yield (0.919), suggest Zn's vital role in productivity. Traits like Plant and Root weights also show high interdependence. Negative correlations, like Soil Fe with Plant Height (-0.865) and Soil Mn with Fruit yield (-0.752), indicate potential toxicity or nutrient imbalance. Notably, low soil nutrient levels alongside high plant uptake may result from enhanced absorption promoted by Trichoderma , which improves nutrient availability. Clustering reveals distinct groups, aiding targeted nutrient and yield management. Conclusion The present study concludes that native Trichoderma isolates were more compatible with agrochemicals than the commercial isolate. Among them, PSV and GMV were the most compatible with the highest number of agrochemicals. The compatibility could be due to the isolates' tolerance to the chemicals, insufficient concentration, or the narrow spectrum of the chemicals, allowing Trichoderma to develop mechanisms to tolerate their toxicity. To study nutrient uptake in tomato, three Trichoderma treatments—T1 (seed treatment), T2 (combined seed and soil application), and T3 (soil application)—along with a control were evaluated under greenhouse conditions. T2 was the most effective, showing significant improvement in growth and nutrient content at 30 DAT. T1 was not significantly superior to T2 or T3 in terms of growth parameters. Among the isolates, SMV and PSV were most effective in enhancing nutrient uptake. SMV improved Fe, Mn, Cu, and Zn uptake, while PSV increased Zn, Cu, and Fe. PSV was the most effective overall in promoting growth and fruit yield, followed by SMV. In this study, Trichoderma spp. applied as both a seed treatment and soil application enhanced plant growth and yield. Known for improving nutrient uptake, especially zinc, Trichoderma promotes root development and increases plant biomass by releasing plant growth hormones like indole-3-acetic acid (IAA). These effects likely contributed to the positive correlations observed between root length, plant height, dry weight, and fruit yield. Additionally, Trichoderma helps mitigate the negative effects of soil-borne pathogens and nutrient imbalances, highlighting its potential as a sustainable biocontrol agent to enhance plant productivity and nutrient efficiency. Integrated Disease Management (IDM) is essential for reducing agrochemical hazards and ensuring healthy crop production. As biological control is a key IDM component, identifying and standardizing efficient BCAs is crucial. In this study, the native isolate PSV ( T. harzianum ) was found effective and will be further evaluated under field conditions for its biocontrol and growth-promoting mechanisms. Declarations Acknowledgement I hereby acknowledge that the data furnished here is the outcome of my research work and is not published or under publish anywhere. No AI tools were used for the preparation of article. Author contribution First author has carried out all the work under the guidance of remaining authors. Conflict of interest All the authors don’t have any conflict in publishing the data and manuscript. All have full interest in publishing the article. Data availability All the data mentioned in the manuscript is own, original and derived from our study, which we can provide to personal as and when required. The data sets analysed during present study are available from first and corresponding author (both same) on reasonable request. Funding declaration All the works were conducted using financial support from the department of Plant Pathology, College of Agriculture, V C Farm, Mandya provide for PG research work and no funding was received from any projects/ agencies/ person. References Abul, H. M., Manjurul, H. M., Amdadul, H. M. and Ilias, G. N. M. (2012). Trichoderma -enriched biofertilizer enhances production and nutritional quality of tomato ( Lycopersicon esculentum Mill.) and minimizes NPK fertilizer use. Agricultural Research, 1 (3): 265-272. Abul, H. M., Manjurul, H. M., Amdadul, H. M. and Ilias, G. N. M. (2012). Trichoderma -enriched biofertilizer enhances production and nutritional quality of tomato ( Lycopersicon esculentum Mill.) and minimizes NPK fertilizer use. Agricultural Research, 1 (3): 265-272. Adams, P., De-leij, F. A. A. M. and Lynch, J. M. (2007). Trichoderma harzianum Rifai 1295-22 mediates growth promotion of crack willow ( Salix fragilis ) saplings in both clean and metal-contaminated soil. Microbiol Ecology, 54: 306-313. Ajith, C. R., Pankaja N. S., Supriya S., Mahadeva J., Umashankar Kumar N., and Mummineni Sunitha. (2024) Compatibility Assessment of Native Non-Rhizospheric Trichoderma Isolates with Various Fungicides. Journal of Advances in Biology & Biotechnology, 27 (9):988-99. Ajith, C. R., Pankaja, N. S., Umashankar Kumar, N., and Mahadev, J., (2023), Non-rhizospheric Trichoderma - a boon for improving nutrient uptake, plant growth and yield in tomato. Biological forum- an International Journal, 15(7): 117-127. Altomare C, Norvell WA, Harman GE, (1999) Trichoderma nutrient solubilisation: Solubilization of phosphates and micronutrients by the plant-growth-promoting and biocontrol fungus Trichoderma harzianum Rifai. Applied Environmental Microbiol ogy, 23: 2926-2933. Bais, H. P., Tiffany, L. W., Laura, G. P., Simon, G. and Jorge, M. V. (2006). The role of root exudates in rhizosphere interactions with plants and other organism’s. Annual review of Plnat Biology, 57:233-266. Bombiti, N., Diana, M. and Puffy, S. (2011). Tomato ( Solanum lycopersicum L.) seedling growth and development as influenced by Trichoderma harzianum and arbuscular mycorrhizal fungi. African Journal of Microbiological Research, 5(4): 425-431. Cuevas, V. C., 2006, Soil inoculation with Trichoderma pseudokoningii Rifai enhances yield of rice. Philippine J. Sci. , 135(1): 31-37 Glic, B. R. (1995). The enhancement of plant growth by free-living bacteria. Canadian Journal of Microbiology, 41:109-117. Harman, G. E. (2006), Overview of mechanisms and uses of Trichoderma spp. Phytopathology, 96(2): 190-4. Harman, G. E. (2011). Multifunctional fungal plant symbionts: new tools to enhance plant growth and productivity. New Phytol ogy, 189: 647–649. Harman, G., Charles, R. H., Ada, V. and Ilan, C. (2004). Trichoderma species- opportunistic, a-virulent plant symbiont. National Reviews of Microbiology, 2(1): 43-56. Hoyos, C. L., Orduz, S., and Bissett, J. (2009). Growth stimulation in bean ( Phaseolus vulgaris L.) by Trichoderma. B iological Control . 51(3): 409–416. Inbar, J., Abramsky, M., Cohen, D. and Chet, I. (1994). Plant growth enhancement and disease control by Trichoderma harzianum in vegetable seedlings grown under commercial conditions. European Journal of Plant. Pathol ogy, 100: 337–346. Jeffries, P., Gianinazzi, S., Perotto, S., Turnau, K. and Barea, J. M. (2003). The contribution of arbuscular mycorrhizal fungi in sustainable maintenance of plant health and soil fertility. Biology and Fertility of Soils, 37: 1-16. Khoshmanzar, E., Aliasgharzad, N., Neyshabouri, M. R., Khoshru, B., Arzanlou, M. and Asgari, L. B. (2020). Effects of Native isolates of Trichoderma spp. on tomato growth and inducing its tolerance to water-deficit stress. International Journal of Environmental Science and Technology, 17: 869-878. Kumar, P., Sharma, D., Singh, R., and Gupta, N. (2023). Iron Deficiency and Its Impact on Plant Growth: A Review . Advances in Plant Nutrition , 18(2), 110-125. Li, X., Zhang, Y., Liu, H., and Wang, L. (2021) . Correlation Between Plant Height and Root Development: Implications for Crop Improvement . Plant Physiology , 29(3), 215-229. Lindsay, W. L. and Norvell, W. A. (1978). Development of DTPA soil test for Zn, Mn, Fe and Cu. Soil Science Society of America J ournal, 42: 421-428. Madhusudhan, P., Gopal, K., Haritha, V., Sangale, U. R. and Rao, S. V. R. K., 2010, Compatibility of Trichoderma viride with fungicides and efficiency against Fusarium solani. J. Pl. Dis. Sci., 5 : 23-26. Mastouri, F., Bjorkman, T. and Harman, G.E. (2010). Seed treatment with Trichoderma harzianum alleviates biotic, abiotic, and physiological stresses in germinating seeds and seedlings. Phytopathology , 100: 1213-1221. Ozbay, N. and Newman, S. E. (2004). Biological control with Trichoderma spp . with emphasis on T. harzianum. Pakistan Journal of Biological Science, 7 (8): 478-484. Page, A. L., Miller, R. H. and Keeney, D. R. (1982). Methods of soil analysis. Part 2- chemical and microbiological properties. 2 nd edition. Agronomy No. 9 Part 2. ASA, SSSA, Madison, Wisconsin, USA. Piper, C. S. (1966). Soil and Plant Analysis , Inter-science Publishers, Inc., New York, pp. 368. Praful, K. and Mane, S. S., 2017, Studies on the compatibility of biocontrol agents with certain fungicides. Int. J. Curr. Microbiol. App. Sci., 6 (3): 1639-1644. Rasool, A., Behzad, H. and Abolfazl, G. (2011). Effect of Native isolates of Trichoderma spp. on tomato seedling growth response and nutrient uptake. African Journal of Biotechnology, 10(31): 5850-5855. Rudresha, D. L., Shivaprakash, M. K. and Prasad, R. D. (2005). Effect of combined application of Rhizobium, phosphate solubilizing bacterium and Trichoderma spp. on growth, nutrient uptake and yield of chickpea ( Cicer aritenium L.). Applied Soil Ecology, 28: 139-146. Sayaji, T. and Prasun, K. (2015). A novel seed-dressing formulation based on an improved mutant strain of Trichoderma virens and its filed evaluation. Frontiers in Microbiology, 25(6): 56-62. Shanmugaiah, V., Balasubramanian, N., Gomathinayagam, S., Manoharan, P. T. and Rajendran A (2009). Effect of single application of Trichoderma viride and Pseudomonas fluorescens on growth promotion in cotton plants. African Journal of Agricultural Research, 4(11): 1220-1225. Sharma, R., Patel, S., Kumar, V., and Singh, A. (2022). Role of Zinc in Enhancing Plant Biomass and Fruit Yield . Journal of Agricultural Science , 14(5), 123-135. Shoresh, M., Harman, G. E. and Mastouri, F. (2010). Induced systemic resistance and plant response to fungal biocontrol agents. Annual Review of Phytopathology, 48:21-43. Singh, S. P., Singh, H. B., Singh, D. K., and Amitava, R. (2014). Trichoderma -mediated enhancement of nutrient uptake and reduction in incidence of Rhizoctonia solani in tomato. Egyptian Journal of Biology , 16: 29-38. Theertha. V. K., Veena, S.S. Karthikeyan, S. and Sreekumar, J., 2017, Compatibility of Trichoderma asperellum with fungicides, insecticides, inorganic fertilizers and bio-pesticides. J. Root Crops , 43(2): 68-75. Thiruchchelvan, N. G., Mikunthan, G. Thirukkumaran and Pakeerathan, K., 2013, Effect of insecticides on bio-agent Trichoderma harzianum rifai under in vitro condition. J. Agri ., 13(10): 1357–1360. Vinale, F., Krishnapillai, S., Emilio, L., Ghisalbertic, Roberta, M., Sheridan, L., Wooa, and Matteo, L., 2008, Trichoderma –plant–pathogen interactions. Soil Biol. Biochem., 40: 1–10. Vincent, J. M., 1927, Distortion of fungal hyphae in the presence of certain inhibitors. Nature., 59 : 850. Wang, Y., Zhang, J., Li, X., and Zhao, M. (2022). Phytotoxic Effects of Excessive Copper and Manganese on Plant Growth and Yield . Environmental Toxicology and Chemistry , 41(7), 2114-2125. Ying, T. L., San, G. H., Yuh, M. H. and Cheng, H. H. (2018). Effects of Trichoderma asperellum on nutrient uptake and Fusarium wilt of tomato, European Journal of Plant Pathol ogy, 129: 112-118. Zhang, J., Wang, H., Li, Y., and Chen, Z. (2021). Toxicity of Manganese and Copper on Plant Development: Mechanisms and Implications for Soil Management . Soil Science Society of America Journal , 85(4), 993-1004. Tables Tables 1 to 7 are available in the Supplementary Files section Plates Plates 1-4 are available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files NUEmicrotablessupplementary.docx Plates.docx Tables.docx Cite Share Download PDF Status: Posted Version 1 posted 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. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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16:47:56","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":104721,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eInfluence of native non-rhizosphere \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eTrichoderma \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eisolates on root growth parameters of tomato\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6653952/v1/5a2c22114d43be1a91c6cd44.png"},{"id":84012566,"identity":"54cc5954-d4c0-4486-aa7e-e05ee2a8ceb8","added_by":"auto","created_at":"2025-06-05 16:47:56","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":163650,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eClustered correlation heat map of micronutrient status v/s tomato plant growth and yield\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6653952/v1/aeb448aa03f8bad4dfb7fe0f.png"},{"id":87545407,"identity":"793375ba-deea-4cc7-9369-90a3e92fc008","added_by":"auto","created_at":"2025-07-25 04:46:54","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1305829,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6653952/v1/648cdff5-1ee3-4c34-9baa-919513932720.pdf"},{"id":84013297,"identity":"4009b5d2-41d7-4e7d-a9c5-3bc3f84875db","added_by":"auto","created_at":"2025-06-05 16:55:56","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":29579,"visible":true,"origin":"","legend":"","description":"","filename":"NUEmicrotablessupplementary.docx","url":"https://assets-eu.researchsquare.com/files/rs-6653952/v1/13e8dcb7f6eec3d170099144.docx"},{"id":84012568,"identity":"1ba286dd-f428-4a2c-bb13-283aa1f8ace6","added_by":"auto","created_at":"2025-06-05 16:47:56","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":4475419,"visible":true,"origin":"","legend":"","description":"","filename":"Plates.docx","url":"https://assets-eu.researchsquare.com/files/rs-6653952/v1/8e12c8421e7e74873f31ab92.docx"},{"id":84012562,"identity":"13ce0621-a687-4654-b08f-42fbc89473c7","added_by":"auto","created_at":"2025-06-05 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These microbes promote plant growth by suppressing phytopathogens, enhancing nutrient uptake and cycling, producing growth hormones, and acting as biocontrol agents (Jeffries \u003cem\u003eet al.\u003c/em\u003e 2003; Glic 1995). Among them, \u003cem\u003eTrichoderma\u003c/em\u003e is a key biocontrol agent widely used in agriculture. It not only combats plant pathogens but also boosts plant growth (Shoresh \u003cem\u003eet al.\u003c/em\u003e 2010), improves poor-quality seed vigour (Mastouri \u003cem\u003eet al.\u003c/em\u003e 2010; Shoresh \u003cem\u003eet al.\u003c/em\u003e 2010), enhances nitrogen-use efficiency (Shoresh \u003cem\u003eet al\u003c/em\u003e. 2010; Harman 2011), and solubilizes micronutrients (Altomare \u003cem\u003eet al.\u003c/em\u003e 1999).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eTrichoderma\u003c/em\u003e spp. is a fungal genus found globally in diverse environments like soil, forests, wood, and paper (Harman \u003cem\u003eet al\u003c/em\u003e., 2004). These fungi form symbiotic relationships with plants, encouraging robust root growth. Widely studied for their ability to enhance plant growth, \u003cem\u003eTrichoderma\u003c/em\u003e species act as biofertilizers and biological control agents by producing antibiotics, parasitizing harmful fungi, and outcompeting plant pathogens (Adams \u003cem\u003eet al.,\u003c/em\u003e 2007; Bais \u003cem\u003eet al.,\u003c/em\u003e 2006). Some species promote growth by solubilizing phosphates and micronutrients like Fe, Mn, and Mg, improving nutrient uptake, and controlling root pathogens (Hoyos \u003cem\u003eet al.,\u003c/em\u003e 2009). They also enhance plant defense against biotic and abiotic stress (Mastouri \u003cem\u003eet al.,\u003c/em\u003e 2010). Recent applications focus on their role as biostimulants for seedling establishment, growth enhancement, and defense induction (Shanmugaiah \u003cem\u003eet al\u003c/em\u003e., 2009). While once attributed solely to indirect effects, it\u0026apos;s now recognized that certain strains directly impact plant development and crop productivity (Harman, 2006).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eTrichoderma\u003c/em\u003e spp. enhance plant growth by solubilizing essential nutrients like Fe, Cu, Mn, Zn, and natural fertilizers such as rock phosphate and pyrite (Altomare \u003cem\u003eet al\u003c/em\u003e., 1999). Their nutrient-solubilizing ability offers a sustainable alternative to chemical fertilizers and pesticides. Since many soil nutrients are poorly soluble, \u003cem\u003eTrichoderma\u003c/em\u003e aids in their availability by secreting organic acids that dissolve minerals and activate nutrients, improving soil nutrient cycling. Their strong colonization expands rhizosphere-soil interaction and boosts secretion of enzymes like sucrase, urease, and phosphatase, enhancing enzyme activity and nutrient uptake. \u003cem\u003eTrichoderma\u003c/em\u003e also decomposes nitrogen compounds into plant-available forms while reducing NO₂ emissions (Jeffries \u003cem\u003eet al\u003c/em\u003e., 2003), and improves nutrient efficiency, lowering the need for nitrogen fertilizers (Harman, 2006). Inoculation with \u003cem\u003eTrichoderma\u003c/em\u003e increases effective nutrient content and soil enzyme activity, supporting soil restoration and plant growth (Glic, 1995).\u003c/p\u003e\n\u003cp\u003eNutrients are vital for crop productivity and are classified as primary, secondary, and micronutrients based on crop requirements. Nutrient use efficiency refers to yield per unit of nutrient supplied from soil or fertilizer (Sayaji and Prasun, 2015). Common limiting nutrients include N, P, K, S, and micronutrients. Their availability is influenced by soil and rhizosphere microorganisms. Microbial enhancement of nutrient use efficiency is increasingly important due to declining soil fertility from intensive agriculture (Sayaji and Prasun, 2015).\u003c/p\u003e\n\u003cp\u003ePreviously, the same isolates were evaluated for compatibility with different fungicides (Ajith \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e2024) and influence on major nutrient uptake and growth promotion in tomato (Ajith \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e2023). In both study we found a significant compatibility and promotion of nutrient uptake and growth promotion. \u0026nbsp;In this study, \u003cem\u003eTrichoderma\u003c/em\u003e spp. were isolated from various local, non-rhizosphere sources and screened against soil-borne plant pathogens using the dual culture technique. Five effective local isolates were identified. To evaluate their impact on plant growth and nutrient use efficiency in tomato, a pot culture experiment was conducted under greenhouse conditions, as discussed below.\u003c/p\u003e"},{"header":"MATERIALS AND METHOD","content":"\u003cp\u003e\u003cstrong\u003eCompatibility with agrochemicals\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFive effective non-rhizosphere \u003cem\u003eTrichoderma\u003c/em\u003e isolates were cultured on sterile PDA medium for 3-4 days. \u003cem\u003eT. viride\u003c/em\u003e, the commercial product, was used as a standard for comparison. Compatibility of the isolates with various agrochemicals (insecticides, bactericides, and herbicides) commonly used in tomato cultivation was tested using the poisoned food technique at concentrations of 50, 100, 200, 400, 600, 1000, and 1500 ppm. The chemicals were mixed with sterilized PDA medium, and mycelial discs of \u003cem\u003eTrichoderma\u003c/em\u003e were inoculated into the plates. The plates were incubated at 28±2°C, and mycelial growth was measured to calculate percent inhibition.\u003c/p\u003e\n\u003cp\u003ePer cent inhibition of \u003cem\u003eTrichoderma\u0026nbsp;\u003c/em\u003ewas calculated by formula as mentioned below given by Vincent (1927).\u003c/p\u003e\n\u003cp\u003eI = (C-T/C) × 100\u003c/p\u003e\n\u003cp\u003eWhere,\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eI = Per cent inhibition\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eC = Radial mycelial growth of \u003cem\u003eTrichoderma\u003c/em\u003e isolates in control\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eT = Radial mycelial growth of \u003cem\u003eTrichoderma\u003c/em\u003e isolates in treatments\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInfluence on nutrient use efficiency in tomato\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFive \u003cem\u003eTrichoderma\u003c/em\u003e isolates and one commercial formulation were evaluated for their effect on nutrient use efficiency (NUE) in tomato under greenhouse conditions using pot culture during the summer of 2020 at the College of Agriculture, V.C. Farm, Mandya. The tomato cultivar 'Abhinav' was used, with treatment combinations detailed in the accompanying table.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMass multiplication of \u003cem\u003eTrichoderma\u0026nbsp;\u003c/em\u003eand quality test\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the \u003cem\u003eTrichoderma\u0026nbsp;\u003c/em\u003eisolates were mass multiplied on PDB media and formulated into talc formulation by mixing in 1:2 ratio (Trichoderma culture: talc powder). Formulation is maintained with 2*10\u003csup\u003e6\u003c/sup\u003e cfu/ml in all the isolates (Plate 2).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSeed treatment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTomato seeds of cultivar Abhinav was treated with jiggery syrup and then with talc formulation @ 4g/kg seeds. Later seeds were shade dried and sown in pro trays to raise seedlings.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSoil application\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe talc formulation was mixed with potting mixture at 10g/kg of soil 10days before transplanting seedlings.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSoil and Plant analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSoil sample was collected before \u003cem\u003eTrichoderma\u0026nbsp;\u003c/em\u003eapplication and after \u003cem\u003eTrichoderma\u0026nbsp;\u003c/em\u003eapplication, just before transplanting tomato seedlings. Tomato plant sample was collected 30 days after transplanting for nutrient analysis. Above ground plant sample was collected and fresh weight was taken using weighing balance. Then the sample was dried in hot air oven at 70±2⁰C for 48 hours. Then plant sample was ground to fine powder using grinding mill and stored for analysis.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSoil analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDTPA – Extractable micronutrients\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMicronutrient cations (Fe, Mn, Zn and Cu) from soil were extracted with DTPA extractant (0.005 M Diethylene Triamine Penta Acetic acid + 0.01 M CaCl\u003csub\u003e2\u003c/sub\u003e + 0.1 M Triethanolamine buffered to pH 7.3) at 1:2 soil to extractant ratio as described by Lindsay and Norvell (1978). The concentration of these cations was determined by atomic absorption spectrophotometer under suitable measuring conditions (Page \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e1982).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDigestion of plant samples with Di-acid mixture\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePowdered plant sample of 1.0 g were pre-digested with conc. HNO\u003csub\u003e3\u003c/sub\u003e overnight and then digested with di-acid mixture containing HNO\u003csub\u003e3\u003c/sub\u003e and HClO\u003csub\u003e4\u003c/sub\u003e in the proportion of 9:4 till a snow white residue was obtained (Piper, 1966). The volume of the digest was made to 100 ml with distilled water and used for total elemental analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePlant analysis for micronutrients (Fe, Mn, Zn and Cu)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMicronutrients \u003cstrong\u003e(\u003c/strong\u003eFe, Mn, Zn and Cu) concentration in the di-acid extract was determined using Atomic Absorption Spectrophotometer (AAS) under suitable measuring conditions (Lindsay and Norwell, 1978; Page \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e1982).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePlant growth and yield\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eObservations\u0026nbsp;were\u0026nbsp;taken on\u0026nbsp;growth\u0026nbsp;parameters\u0026nbsp;of\u0026nbsp;tomato\u0026nbsp;like\u0026nbsp;plant\u0026nbsp;height,\u0026nbsp;plant\u0026nbsp;fresh\u0026nbsp;and\u0026nbsp;dry\u0026nbsp;weight, root length, root fresh and dry weight. Tomato fruit was harvested regularly starting\u0026nbsp;from\u0026nbsp;75 DAT\u0026nbsp;and\u0026nbsp;yield\u0026nbsp;from different\u0026nbsp;treatments\u0026nbsp;was compared\u0026nbsp;with control.\u003c/p\u003e"},{"header":"Experimental results","content":"\u003cp\u003e\u003cstrong\u003eCompatibility with insecticides:\u0026nbsp;\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAmong seven insecticides (Table 1), chlorpyrifos 50% EC caused minimal mycelial inhibition (Plate 1) (\u0026lt;10%) up to 600 ppm in native \u003cem\u003eTrichoderma\u003c/em\u003e isolates, with GMV showing the least inhibition (0\u0026ndash;8.25%). The commercial isolate showed the highest inhibition (3\u0026ndash;49.33%) across concentrations. Similarly, thiamethoxam 25% WG caused \u0026lt;5% inhibition up to 400 ppm, with PSV showing the least (20.35%) and the commercial isolate the highest (51.24%) inhibition at 1500 ppm. Flubendiamide 20% WG showed no mycelial inhibition up to 200 ppm in SMV, SDKd, GMV, and CPV isolates. At 1500 ppm, inhibition ranged from 16.38% to 68.36%, with GMV showing the least and PSV the highest inhibition.\u003cstrong\u003e\u003cbr\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFipronil 5% SC was compatible with all \u003cem\u003eTrichoderma\u003c/em\u003e isolates, showing no inhibition up to 400 ppm in SDKd, GMV, and PSV. At 1500 ppm, these isolates showed low inhibition (13.65\u0026ndash;33.85%), while CPV and the commercial isolate showed moderate inhibition (62.33\u0026ndash;67.88%). Chlorantraniliprole 20% SC also showed compatibility, with GMV showing least inhibition (0\u0026ndash;11.54%) and the commercial isolate the highest (21.85\u0026ndash;67.63%). At 1500 ppm, SDKd and GMV remained highly compatible.\u003c/p\u003e\n\u003cp\u003eEmamectin benzoate 5% SG showed good compatibility, with GMV, SDKd, and PSV displaying 0% inhibition up to 200 ppm and \u0026lt;40% at 1500 ppm. Other isolates, including the commercial one, showed moderate incompatibility (\u0026lt;70% inhibition). Imidacloprid 20% SC caused the least inhibition in GMV (0\u0026ndash;50.44%) and the highest in the commercial isolate (0\u0026ndash;58.33%). At 1500 ppm, it was moderately incompatible with all \u003cem\u003eTrichoderma\u003c/em\u003e isolates.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompatibility with bactericide and herbicide:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo assess compatibility, one bactericide and one herbicide were tested on native \u003cem\u003eTrichoderma\u003c/em\u003e isolates at seven concentrations (Table 2). Both were compatible. The herbicide butachlor showed no mycelial inhibition at 50 and 100 ppm in all isolates, with GMV showing 0% inhibition up to 600 ppm. At 1500 ppm, GMV showed the least inhibition (13.52%) and the commercial isolate the highest (58.31%).\u003c/p\u003e\n\u003cp\u003eFor the bactericide K-cyclin, SDKd and GMV showed 0% mycelial inhibition up to 200 ppm, while PSV and SMV showed no inhibition up to 100 ppm. The bactericide remained compatible up to 1000 ppm in most isolates, except PSV, CPV, and the commercial isolate, which showed moderate inhibition (51.24\u0026ndash;63.37%). At 1500 ppm, SDKd had the least inhibition (41.23%) and PSV the highest (68.32%), with the commercial isolate showing 67.65%.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe compatibility of seven insecticides, one herbicide, and one bactericide with native \u003cem\u003eTrichoderma\u003c/em\u003e isolates and a commercial isolate was tested. All agrochemicals were compatible with varying mycelial inhibition. Chlorpyrifos 50% EC was the most compatible, followed by thiamethoxam 25% WG and flubendiamide 20% WG. Imidacloprid 20% SC and emamectin benzoate 5% SG were the least compatible, showing moderate incompatibility at 1500 ppm.\u003c/p\u003e\n\u003cp\u003ePraful and Mane (2017) observed that chlorantraniliprole 20% SC, emamectin benzoate 5% SG, and chlorpyrifos 50% EC were highly compatible with \u003cem\u003eT. pseudokoningii\u003c/em\u003e at 10-2000 ppm. Similarly, chlorantraniliprole 20% SC and imidacloprid 20% SC were compatible with \u003cem\u003eT. harzianum\u003c/em\u003e, with 85mm colony growth, while thiamethoxam 25% WG showed 3.82% mycelial inhibition (Thiruchchelvan \u003cem\u003eet al.,\u003c/em\u003e 2013). Madhusudhan \u003cem\u003eet al\u003c/em\u003e. (2010) found that thiamethoxam 25% WG and fipronil 5% SC were compatible with \u003cem\u003eT. viride\u003c/em\u003e, with 12% and 14.3% inhibition. Theertha \u003cem\u003eet al.\u003c/em\u003e (2017) noted that chlorpyrifos, fipronil, and flubendiamide were compatible with \u003cem\u003eT. asperellum\u003c/em\u003e with less than 15% inhibition up to 800 ppm.When compared among all the agrochemicals, SDKd, GMV and PSV were the isolates which showed compatibility with most number of agrochemicals, followed by SMV and CPV. However, the commercial isolate showed compatibility with least number of agrochemicals (Supplementary table 1 \u0026amp; 2).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe isolate SDKd showed high compatibility with insecticides chlorpyrifos, emamectin benzoate, imidacloprid, thiamethoxam, flubendiamide, chlorantraniliprole, fipronil, and the herbicide butachlor, and moderate compatibility with bactericide K-cycline. It showed moderate incompatibility with tebuconazole and difenconazole. The GMV isolate was highly compatible with chlorpyrifos, flubendiamide, butachlor, and bactericide K-cycline. It was also compatible with emamectin benzoate, fipronil, thiamethoxam, and chlorantraniliprole, with moderate compatibility with imidacloprid. PSV showed high compatibility with chlorpyrifos, fipronil, and butachlor, with moderate compatibility with thiamethoxam, emamectin benzoate, and chlorantraniliprole, while moderate incompatibility was seen with flubendiamide and K-cycline. CPV exhibited high compatibility with chlorpyrifos and butachlor, and moderate compatibility with thiamethoxam, emamectin benzoate, chlorantraniliprole, and imidacloprid. SMV showed high compatibility with thiamethoxam and butachlor, and compatibility with chlorantraniliprole, fipronil, flubendiamide, imidacloprid, chlorpyrifos, emamectin benzoate, and K-cycline. The commercial isolate was compatible with chlorpyrifos, thiamethoxam, flubendiamide, emamectin benzoate, imidacloprid, chlorantraniliprole, and butachlor, with moderate compatibility with fipronil and K-cycline.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInfluence on nutrient use efficiency in tomato\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGermination percentage:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTomato seeds (variety Abhinav) treated with different native \u003cem\u003eTrichoderma\u003c/em\u003e isolates were sown in pro-trays and monitored for germination. All \u003cem\u003eTrichoderma\u003c/em\u003e treatments significantly improved germination compared to the control (T4). Isolates GMV and PSV showed the highest germination rates at 92.85% and 90%, respectively, followed by SMV (90%). SDKd had the lowest among native isolates (88.57%), while the commercial isolate recorded the lowest overall (85.71%).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNutrient status of soil before \u003cem\u003eTrichoderma\u0026nbsp;\u003c/em\u003etreatment\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTotally four micro nutrients \u003cem\u003eviz.,\u003c/em\u003e DTPA exchangeable Zn, Cu, Mn and Fe were assessed in the laboratory using standard protocols (Table 3). The soil contained 3.10 mgkg\u003csup\u003e-1\u0026nbsp;\u003c/sup\u003eof Zn, 1.39 mgkg\u003csup\u003e-1\u0026nbsp;\u003c/sup\u003eof copper (Cu), 5.86 mgkg\u003csup\u003e-1\u003c/sup\u003e of manganese (Mn) and 9.13 mgkg\u003csup\u003e-1\u003c/sup\u003e of iron (Fe).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNutrient status of soil after \u003cem\u003eTrichoderma\u0026nbsp;\u003c/em\u003etreatment:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eRhizosphere soil analysis 30 days post-transplanting showed increased nutrient content after \u003cem\u003eTrichoderma\u003c/em\u003e treatment compared to pre-treatment levels (Table 4).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eZinc (Zn) (mgkg\u003csup\u003e-1\u0026nbsp;\u003c/sup\u003eof soil):\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSoil Zn content varied significantly across isolates, treatments, and their interactions. In T1, SDKd and commercial isolates showed the highest Zn (2.90 mgkg⁻\u0026sup1;), while CPV recorded the lowest (2.50 mgkg⁻\u0026sup1;). In T2, SMV and GMV had the highest Zn (2.70 mgkg⁻\u0026sup1;), and CPV the lowest (2.30 mgkg⁻\u0026sup1;). In T3, SMV showed the highest Zn (2.90 mgkg⁻\u0026sup1;), followed by commercial (2.70 mgkg⁻\u0026sup1;), while CPV and GMV had the lowest (2.40 mgkg⁻\u0026sup1;).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCopper (Cu)\u003c/strong\u003e \u003cstrong\u003e(mgkg\u003csup\u003e-1\u0026nbsp;\u003c/sup\u003eof soil):\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCu content varied across treatments and isolates. In T1, commercial and SMV showed highest Cu (1.35 and 1.33 mgkg⁻\u0026sup1;), while GMV and CPV had the lowest (1.25 and 1.27 mgkg⁻\u0026sup1;). In T2, commercial and SMV again led (1.33 and 1.30 mgkg⁻\u0026sup1;), with GMV, CPV, and PSV lowest (1.22\u0026ndash;1.25 mgkg⁻\u0026sup1;). In T3, SMV had highest Cu (1.31 mgkg⁻\u0026sup1;), followed by commercial (1.30 mgkg⁻\u0026sup1;), while GMV and CPV showed lowest (1.23\u0026ndash;1.24 mgkg⁻\u0026sup1;).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eManganese (mgkg\u003csup\u003e-1\u0026nbsp;\u003c/sup\u003eof soil):\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMn content differed among isolates and treatments. In T1, commercial and SDKd showed highest Mn (5.71 and 5.68 mgkg⁻\u0026sup1;), while GMV had the lowest (5.61 mgkg⁻\u0026sup1;). PSV and CPV recorded 5.62 mgkg⁻\u0026sup1;. In T2, commercial and SDKd again led (5.68 and 5.65 mgkg⁻\u0026sup1;), with GMV lowest (5.57 mgkg⁻\u0026sup1;). SMV, PSV, and CPV followed at 5.60 mgkg⁻\u0026sup1;. In T3, commercial (5.65 mgkg⁻\u0026sup1;) and SMV (5.63 mgkg⁻\u0026sup1;) had highest Mn, while SDKd, GMV, and CPV recorded the lowest (5.60 mgkg⁻\u0026sup1;).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIron (Fe) (mgkg\u003csup\u003e-1\u0026nbsp;\u003c/sup\u003eof soil):\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFe content varied across treatments. In T1, commercial and CPV recorded highest Fe (9.01 and 8.97 mgkg⁻\u0026sup1;), while GMV had the lowest (8.85 mgkg⁻\u0026sup1;). In T2, commercial (9.03 mgkg⁻\u0026sup1;) and CPV (8.95 mgkg⁻\u0026sup1;) again showed higher Fe levels, with SMV and GMV the lowest (8.87 mgkg⁻\u0026sup1;). In T3, CPV and commercial both recorded 8.97 mgkg⁻\u0026sup1;, while SMV and GMV had the lowest Fe (8.81 mgkg⁻\u0026sup1;).\u003c/p\u003e\n\u003cp\u003eNo significant nutrient differences were noted across treatments and isolates, likely due to enhanced availability and uptake following \u003cem\u003eTrichoderma\u003c/em\u003e application.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNutrient status of tomato plants 30 days after transplanting\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTomato plant analysis showed significant nutrient differences between \u003cem\u003eTrichoderma\u003c/em\u003e treatments and control. Four micronutrients \u003cem\u003eviz.,\u0026nbsp;\u003c/em\u003eZn, Cu, Mn, and Fe were examined and discussed (Table 5; Supplementary table 3).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eZinc (Zn):\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eZn content in tomato tissues (mgkg⁻\u0026sup1;) was highest in T2, with GMV recording 30.94 and CPV the lowest at 22.81. The commercial isolate showed 22.90. In T1, GMV and CPV recorded 24.57 and 21.46, respectively, while the control showed the lowest Zn content at 17.86.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCopper (Cu)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eT2 showed highest Cu content, with PSV and commercial isolates recording 18.60 and 22.90 mgkg⁻\u0026sup1;, respectively. In T1 and T3, PSV had the highest Cu (12.63 and 15.51 mgkg⁻\u0026sup1;), while the control recorded 10.25 mgkg⁻\u0026sup1;.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eManganese (Mn)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eT2 showed the highest Mn content, with SMV and commercial isolates recording 84.23 and 79.56 mgkg⁻\u0026sup1;, respectively. In T1 and T3, the highest Mn was in SDKd (68.35 mgkg⁻\u0026sup1;) and SMV (80.62 mgkg⁻\u0026sup1;). The control showed 57.74 mgkg⁻\u0026sup1;.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIron (Fe):\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn T2, GMV and commercial isolates recorded the highest Fe content (172.36 and 169.62 mgkg⁻\u0026sup1;). In T1 and T3, GMV and PSV had the highest Fe levels (131.23 and 170.25 mgkg⁻\u0026sup1;), while the control recorded 121.36 mgkg⁻\u0026sup1;.\u003c/p\u003e\n\u003cp\u003eSimilar findings were reported by Rudresh \u003cem\u003eet al.\u003c/em\u003e (2005), who observed that native isolates of \u003cem\u003eTrichoderma\u003c/em\u003e spp. were capable of solubilizing insoluble tricalcium phosphate to varying degrees. In pot culture experiments using rock phosphate, enhanced phosphorus uptake was recorded in plants treated with \u003cem\u003eT. harzianum\u003c/em\u003e (PDBCTH 10), followed by \u003cem\u003eT. virens\u003c/em\u003e (PDBCTVs 12) and \u003cem\u003eT. viride\u003c/em\u003e (TV 97). Rasool \u003cem\u003eet al.\u003c/em\u003e (2011) found that tomato plants grown in soil fortified with \u003cem\u003eTrichoderma\u003c/em\u003e spp. T and \u003cem\u003eT. harzianum\u003c/em\u003e T969 exhibited increased uptake of nutrients such as nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg), with soil application proving more effective than seed treatment. Bombiti \u003cem\u003eet al\u003c/em\u003e. (2011) reported that pre-sowing application of \u003cem\u003eT. harzianum\u003c/em\u003e alone led to improved shoot uptake of N, P, Ca, Mg, and sulfur (S). Under glasshouse conditions, Inbar \u003cem\u003eet al\u003c/em\u003e. (1994) demonstrated that \u003cem\u003eTrichoderma\u003c/em\u003e significantly increased phosphorus and potassium content in tomato leaves even without fertilizer application. Ying \u003cem\u003eet al.\u003c/em\u003e (2018) noted that the \u003cem\u003eT. asperellum\u003c/em\u003e CHF 78 isolate, which produces enzymes such as cellulases, chitinases, proteases, and siderophores along with indole acetic acid (IAA), substantially enhanced the uptake of P, K, Mg, and Zn. Similarly, Singh \u003cem\u003eet al\u003c/em\u003e. (2014) observed that a combination of \u003cem\u003eT. harzianum\u003c/em\u003e isolates (BHU51 + BHU105) increased the mineral content of tomato plants\u0026mdash;N, P, K, Ca, Mg, S, Zn, Cu, Mn, and Fe\u0026mdash;by 40\u0026ndash;50% compared to the untreated control.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTomato\u0026nbsp;growth\u0026nbsp;promotion\u0026nbsp;by \u003cem\u003eTrichoderma\u0026nbsp;\u003c/em\u003etreatment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTomato growth parameters \u003cem\u003eviz.,\u0026nbsp;\u003c/em\u003eplant height, fresh and dry weights of plant and root, root length, and fruit yield were recorded at intervals to assess how Trichoderma isolates affect growth and nutrient uptake from soil (Plate 3 and 4).\u003c/p\u003e\n\u003cp\u003ePlant\u0026nbsp;growth\u0026nbsp;parameters:\u003c/p\u003e\n\u003cp\u003eTomato growth parameters such as plant height (cm) at 15, 30, 45, and 90 DAT and plant fresh/dry weight at 90 DAT showed significant differences among isolates and treatments (Supplementary table 4). The combination treatment (T2) was most effective. At 15 DAT, isolate GMV gave the highest plant height (43.22 cm), followed by PSV (42.68 cm) and CPV (42.53 cm), with GMV and PSV maintaining superior heights at 90 DAT (173.36 cm and 172.32 cm)\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003ecompared to the control (30 cm at 15 DAT, 111.67 cm at 90 DAT). The commercial isolate performed poorest. In T1 and T3, isolate SMV was best at 90 DAT. Trichoderma notably enhanced plant height via improved nutrient uptake.\u003c/p\u003e\n\u003cp\u003eAt crop maturity, plant fresh and dry weight data (Supplementary table 5; Fig. 1) showed that the combined seed and soil treatment (T2) was most effective. Isolate GMV yielded the highest fresh (263.68g) and dry weight (53.22g), followed by PSV (227.69g and 43.68g). The commercial isolate showed lower values (195.66g and 37.68g), and the control was significantly inferior (142.66g and 22.75g). In T1 and T3, GMV again outperformed others (196.56g and 245.68g fresh weight; 29.56g and 40.88g dry weight), while the commercial isolate recorded lower values (144.69g and 187.58g fresh; 23.15g and 28.22g dry).\u003c/p\u003e\n\u003cp\u003eRoot growth parameters\u003c/p\u003e\n\u003cp\u003eIn order to take observations on root length, root fresh and dry weight, the plants were\u0026nbsp;uprooted at maturity of the crop (90DAT) and observed for various\u0026nbsp;parameters (Supplementary table 6; Fig. 2; Plate 4). With respect to all parameters, T2\u0026nbsp;was significantly superior over other treatments and control.\u003c/p\u003e\n\u003cp\u003eIn T2, SMV was the top-performing isolate with 30.45 cm root length, 46.68 g fresh weight, and 14.26 g dry weight, followed by PSV (30.11 cm, 42.12 g, and 12.41 g). The commercial isolate recorded lower values (26.24 cm, 26.77 g, and 7.29 g), while the control had the least (17.88 cm, 15.65 g, and 3.33 g). In T1, GMV had the longest roots (23.13 cm), while PSV showed the highest root fresh and dry weights (21.10 g and 4.54 g). In T3, SMV performed best (28.11 cm, 36.14 g, and 9.78 g).\u003c/p\u003e\n\u003cp\u003eTomato fruit yield\u003c/p\u003e\n\u003cp\u003eTrichoderma-enhanced nutrient uptake led to improved tomato growth and yield. Yield analysis (Table) showed that both treatments and isolates had significant effects. The combination treatment (T2) was most effective, with GMV yielding 521.62 g per plant, followed by PSV (512.45 g), outperforming the commercial isolate (481.23 g), while the control had the lowest yield (318.27 g). In T1, GMV gave the highest yield (367.63 g), and in T3, GMV and PSV performed similarly (484.65 g and 484.38 g, respectively).\u003c/p\u003e\n\u003cp\u003eSimilar findings were reported by Rasool et al. (2011), who observed significant increases in shoot length and shoot/root fresh and dry weights in Trichoderma-fortified soils. Khoshmanzar et al. (2020) found that \u003cem\u003eT. asperellum\u003c/em\u003e B1092 enhanced tomato shoot and root growth over the control. Treatments with \u003cem\u003eT. longibrachiatum\u003c/em\u003e KH (T1), MA (T2), and \u003cem\u003eT. harzianum\u003c/em\u003e (T3) increased shoot dry weight by 97.2%, 17.2%, and 18.96% over the negative control, and by 32.81%, 19.41%, and 20.42% over the positive control.\u003c/p\u003e\n\u003cp\u003eOzbay and Newman (2004) tested \u003cem\u003eT. harzianum\u003c/em\u003e strains (Plant Shield, T22, T95) on 18-day-old tomato seedlings. After six weeks, seedling emergence was 100%, 83.33%, 88.89%, and 83% for Plant Shield, T22, T95, and control, respectively. T95 showed the highest plant height (34.35 cm) and the greatest shoot and root weight (43.10 g, 4.68 g, 9.13 g, and 0.87 g).\u003c/p\u003e\n\u003cp\u003eGreenhouse results showed that \u003cem\u003eT. pseudokoningii\u003c/em\u003e Rifai enhanced nutrient availability in soil (K, Ca, P, Zn, Fe) and in tomato plant tissue. Soil levels were 0.750 cmol/kg K, 10.05 cmol/kg Ca, 41.5 ppm P, 22 ppm Zn, and 12 ppm Fe. Tomato tissues contained 2.195% N, 0.046% P, 2.290% Ca, 0.0075% Zn, and 0.035% Fe. These nutrients promoted higher dry matter production (14g in control vs. 9g in Trichoderma treatment), resulting in earlier flowering and better fruit yield (Cuevas, 2006).\u003c/p\u003e\n\u003cp\u003eAbul \u003cem\u003eet al\u003c/em\u003e. (2012) found that treating tomato seeds with native \u003cem\u003eTrichoderma\u003c/em\u003e isolates (T85, M14, M2_14) significantly increased plant height (M2_14: 25.22 cm, T85: 28.28 cm vs. 22.8 cm control) and root length (M14: 10.5 cm, T85: 10.85 cm vs. 10.05 cm control). \u003cem\u003eT85\u003c/em\u003e also enhanced root fresh/dry weight (4.45g/2.93g vs. 1.91g/0.91g control). Vinale \u003cem\u003eet al.\u003c/em\u003e (2008) found that rhizosphere-competent \u003cem\u003eTrichoderma\u003c/em\u003e isolates produce diffusible metabolites that act as plant growth regulators (auxins or auxin-like compounds), enhancing the growth of colonized plants.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCorrelating nutrient use efficiency with plant growth and yield\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe correlation analysis revealed significant relationships between soil and plant nutrient concentrations and various growth and yield parameters (Table 7; Fig. 3). Notably, plant zinc concentration showed a strong positive correlation with fruit yield (r = 0.919), plant dry weight (r = 0.938), and plant fresh weight (r = 0.878), suggesting that higher zinc accumulation in plant tissue is closely associated with increased biomass production and fruit output. Similarly, plant height exhibited strong positive correlations with root length (r = 0.927) and root dry weight (r = 0.827), indicating that taller plants tend to develop more extensive root systems. On the other hand, soil copper, manganese, and iron levels displayed negative correlations with most growth traits and yield. Specifically, soil copper and manganese had strong negative correlations with fruit yield (r = -0.752 for both), while soil iron showed a pronounced negative association with plant height (r = -0.865), implying that elevated levels of these micronutrients in soil might hinder plant development and productivity. These findings highlight the importance of maintaining optimal micronutrient balance in the soil to ensure healthy plant growth and maximize yield potential.\u003c/p\u003e\n\u003cp\u003eCorrelation analysis showed that plant zinc levels were strongly linked to dry weight (r = 0.938), fresh weight (r = 0.878), and fruit yield (r = 0.919), reinforcing its role in enhancing biomass and productivity (Sharma \u003cem\u003eet al\u003c/em\u003e., 2022). Plant height correlated positively with root length (r = 0.927) and root dry weight (r = 0.827), as noted by Li \u003cem\u003eet al\u003c/em\u003e. (2021). In contrast, soil copper and manganese negatively affected fruit yield (r = -0.752) and plant height (r = -0.656), aligning with reports of phytotoxicity (Wang \u003cem\u003eet al\u003c/em\u003e., 2022; Zhang \u003cem\u003eet al\u003c/em\u003e., 2021). Soil iron also showed a strong negative correlation with plant height (r = -0.865), supporting findings by Kumar \u003cem\u003eet al\u003c/em\u003e. (2023). These patterns highlight the critical role of balanced micronutrient availability in supporting optimal plant growth and yield.\u003c/p\u003e\n\u003cp\u003eThe clustered correlation heat map highlights key relationships between soil nutrients, plant traits, and yield. Strong positive correlations, such as between Plant Zn and both Plant dry weight (0.938) and Fruit yield (0.919), suggest Zn\u0026apos;s vital role in productivity. Traits like Plant and Root weights also show high interdependence. Negative correlations, like Soil Fe with Plant Height (-0.865) and Soil Mn with Fruit yield (-0.752), indicate potential toxicity or nutrient imbalance. Notably, low soil nutrient levels alongside high plant uptake may result from enhanced absorption promoted by \u003cstrong\u003eTrichoderma\u003c/strong\u003e, which improves nutrient availability. Clustering reveals distinct groups, aiding targeted nutrient and yield management.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe present study concludes that native Trichoderma isolates were more compatible with agrochemicals than the commercial isolate. Among them, PSV and GMV were the most compatible with the highest number of agrochemicals. The compatibility could be due to the isolates' tolerance to the chemicals, insufficient concentration, or the narrow spectrum of the chemicals, allowing Trichoderma to develop mechanisms to tolerate their toxicity.\u003c/p\u003e \u003cp\u003eTo study nutrient uptake in tomato, three \u003cem\u003eTrichoderma\u003c/em\u003e treatments\u0026mdash;T1 (seed treatment), T2 (combined seed and soil application), and T3 (soil application)\u0026mdash;along with a control were evaluated under greenhouse conditions. T2 was the most effective, showing significant improvement in growth and nutrient content at 30 DAT. T1 was not significantly superior to T2 or T3 in terms of growth parameters. Among the isolates, SMV and PSV were most effective in enhancing nutrient uptake. SMV improved Fe, Mn, Cu, and Zn uptake, while PSV increased Zn, Cu, and Fe. PSV was the most effective overall in promoting growth and fruit yield, followed by SMV.\u003c/p\u003e \u003cp\u003eIn this study, \u003cem\u003eTrichoderma\u003c/em\u003e spp. applied as both a seed treatment and soil application enhanced plant growth and yield. Known for improving nutrient uptake, especially zinc, \u003cem\u003eTrichoderma\u003c/em\u003e promotes root development and increases plant biomass by releasing plant growth hormones like indole-3-acetic acid (IAA). These effects likely contributed to the positive correlations observed between root length, plant height, dry weight, and fruit yield. Additionally, \u003cem\u003eTrichoderma\u003c/em\u003e helps mitigate the negative effects of soil-borne pathogens and nutrient imbalances, highlighting its potential as a sustainable biocontrol agent to enhance plant productivity and nutrient efficiency.\u003c/p\u003e \u003cp\u003eIntegrated Disease Management (IDM) is essential for reducing agrochemical hazards and ensuring healthy crop production. As biological control is a key IDM component, identifying and standardizing efficient BCAs is crucial. In this study, the native isolate PSV (\u003cem\u003eT. harzianum\u003c/em\u003e) was found effective and will be further evaluated under field conditions for its biocontrol and growth-promoting mechanisms.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eI hereby acknowledge that the data furnished here is the outcome of my research work and is not published or under publish anywhere. No AI tools were used for the preparation of article.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFirst author has carried out all the work under the guidance of remaining authors.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the authors don’t have any conflict in publishing the data and manuscript. All have full interest in publishing the article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the data mentioned in the manuscript is own, original and derived from our study, which we can provide to personal as and when required. The data sets analysed during present study are available from first and corresponding author (both same) on reasonable request.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the works were conducted using financial support from the department of Plant Pathology, College of Agriculture, V C Farm, Mandya provide for PG research work and no funding was received from any projects/ agencies/ person.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbul, H. M., Manjurul, H. M., Amdadul, H. M. and Ilias, G. N. M. 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Agri\u003c/em\u003e., 13(10): 1357\u0026ndash;1360.\u003c/li\u003e\n\u003cli\u003eVinale, F., Krishnapillai, S., Emilio, L., Ghisalbertic, Roberta, M., Sheridan, L., Wooa, and Matteo, L., 2008, \u003cem\u003eTrichoderma\u003c/em\u003e\u0026ndash;plant\u0026ndash;pathogen interactions. \u003cem\u003eSoil Biol. Biochem.,\u003c/em\u003e 40: 1\u0026ndash;10.\u003c/li\u003e\n\u003cli\u003eVincent, J. M., 1927, Distortion of fungal hyphae in the presence of certain inhibitors. \u003cem\u003eNature., \u003c/em\u003e59\u003cstrong\u003e: \u003c/strong\u003e850.\u003c/li\u003e\n\u003cli\u003eWang, Y., Zhang, J., Li, X., and Zhao, M. (2022). \u003cem\u003ePhytotoxic Effects of Excessive Copper and Manganese on Plant Growth and Yield\u003c/em\u003e\u003cem\u003e.\u003c/em\u003e \u003cem\u003eEnvironmental Toxicology and Chemistry\u003c/em\u003e, 41(7), 2114-2125.\u003c/li\u003e\n\u003cli\u003eYing, T. L., San, G. H., Yuh, M. H. and Cheng, H. H. (2018). Effects of \u003cem\u003eTrichoderma asperellum\u003c/em\u003e on nutrient uptake and Fusarium wilt of tomato, \u003cem\u003eEuropean Journal of Plant Pathol\u003c/em\u003eogy, 129: 112-118.\u003c/li\u003e\n\u003cli\u003eZhang, J., Wang, H., Li, Y., and Chen, Z. (2021). \u003cem\u003eToxicity of Manganese and Copper on Plant Development: Mechanisms and Implications for Soil Management\u003c/em\u003e. \u003cem\u003eSoil Science Society of America Journal\u003c/em\u003e, 85(4), 993-1004.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTables 1 to 7 are available in the Supplementary Files section\u003c/p\u003e"},{"header":"Plates","content":"\u003cp\u003ePlates 1-4 are available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Trichoderma, native, non-rhizosphere, compatibility, micro-nutrient, growth and yield","lastPublishedDoi":"10.21203/rs.3.rs-6653952/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6653952/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study investigates the compatibility of native non-rhizosphere Trichoderma isolates with agrochemicals and their impact on nutrient uptake and plant growth in tomato. Native isolates, particularly PSV and GMV, exhibited higher compatibility with agrochemicals compared to a commercial isolate, potentially due to their tolerance to chemical toxicity or the specific nature of the agrochemicals. A greenhouse experiment was conducted to assess the effects of three Trichoderma treatments\u0026mdash;seed treatment (T1), combined seed and soil application (T2), and soil application (T3)\u0026mdash;on tomato growth and nutrient content. T2 was the most effective treatment, leading to significant improvements in growth and nutrient uptake, particularly in iron (Fe), manganese (Mn), copper (Cu), and zinc (Zn), with PSV and SMV isolates being the most beneficial. Among these, PSV notably enhanced growth and fruit yield. The results highlight the potential of native non-rhizosphere Trichoderma spp. to promote nutrient uptake, improve plant biomass, and mitigate the adverse effects of soil-borne pathogens, suggesting their role as a sustainable biocontrol agent. Furthermore, the study emphasizes the importance of Integrated Disease Management (IDM) in reducing agrochemical reliance, with native Trichoderma isolates, especially PSV, identified as promising candidates for future field trials focused on both biocontrol and growth enhancement.\u003c/p\u003e","manuscriptTitle":"Soil-Free Origins, Root-Deep Impact: Unlocking the Power of Native, Non-rhizosphere TrichodermaIsolates in Plant Growth and Yield under Agrochemical Environments","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-05 16:47:51","doi":"10.21203/rs.3.rs-6653952/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4c6f3755-bf06-4862-9868-53150e708f14","owner":[],"postedDate":"June 5th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":49599240,"name":"Biological sciences/Microbiology"},{"id":49599241,"name":"Biological sciences/Plant sciences"},{"id":49599242,"name":"Earth and environmental sciences/Environmental sciences"}],"tags":[],"updatedAt":"2025-07-25T04:38:47+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-05 16:47:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6653952","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6653952","identity":"rs-6653952","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}