Exploring the potential of Trichoderma asperellum TCS007 on growth promotion of pecan seedlings as well as rhizosphere soil nutrients and microbial community

Exploring the potential of Trichoderma asperellum TCS007 on growth promotion of pecan seedlings as well as rhizosphere soil nutrients and microbial community | 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 Exploring the potential of Trichoderma asperellum TCS007 on growth promotion of pecan seedlings as well as rhizosphere soil nutrients and microbial community Hao Cao, Xuesong Li, Hao Han, Sai Chen, Jing Jin, Jing Yuan, Chizhou Liang, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5756661/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 14 May, 2025 Read the published version in Plant and Soil → Version 1 posted 5 You are reading this latest preprint version Abstract Aims Pecan ( Carya cathayensis Sarg.) is an important forest trees in China, the application of chemical pesticides for disease control has caused severe damage to the soil, including reduced fertility and disruption of microbial communities. Although Trichoderma treatment has been shown to promote plant growth and improve soil quality, its effects on the growth promotion of pecan and the impact on soil microbial communities and physicochemical properties remained unclear. Methods In this study, we investigated the impact of T. asperellum TCS007 spore suspension and its fermented crude extract on the growth and development of pecan seedlings. We also explored the effects of TCS007 treatment on the nutrients, enzyme activities, and microbial diversity in the rhizosphere soil of pecan seedlings during their three main growth stages. Results Treatment with TCS007 spore suspension or crude extract promoted the growth of pecan seedlings, with significantly higher levels of leaf hormones and defense enzyme activity compared to the control (CK). Moreover, the content of soil organic matter and ammonium nitrogen, as well as the activity of soil enzymes such as catalase and urease, were all significantly higher than CK after treatment, and the soil pH shifted from slightly acidic to slightly alkaline. The results indicated that TCS007 treatment significantly increased the richness of beneficial fungi and bacteria in the soil. Conclusion The results demonstrated that TCS007 treatment significantly promoted the growth of pecan plants, increased enzyme activity and nutrient content in the soil, and improved the soil micro-ecological environment. Biofertilizers Forest trees Soil physicochemical properties soil enzyme activities microbial diversity Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Introduction Trichoderma spp., ubiquitously present in nature, are a group of fungi that significantly promote plant growth (Chacón et al. 2007 ; Tyśkiewicz et al. 2022 ; Chen et al. 2023 ). These fungi colonize the rhizosphere of plants, enhancing their tolerance to biotic and abiotic stresses (Vargas et al. 2009 ). They produce a variety of bioactive substances, including antibiotics and enzymes, which inhibit plant pathogens. By competing for nutrients and occupying space, they effectively suppress the growth of soil-borne pathogens, thereby protecting plants from disease (Garnica-Vergara et al. 2015 ; Shi et al. 2016 ). Trichoderma harzianum has been shown to grow 2.0 to 4.2 times faster than Botrytis cinerea , thereby effectively inhibiting the latter's growth. This competitive exclusion not only deprives pathogens of essential resources but also physically blocks their access to plant tissues (Yao et al. 2023 ). Moreover, Trichoderma induces systemic resistance in plants, strengthening their resilience to adverse conditions (Kashyap et al. 2017 ; Gupta and Maya 2020 ). These functions make Trichoderma an important biocontrol agent, widely used in agriculture. Trichoderma spp., have a significant positive impact on soil health. They promote the decomposition of organic matter in the soil, enhancing soil fertility and structure (Khan et al. 2017 ; Halifu et al. 2019 ). The metabolic activities of Trichoderma increase the availability of nutrients such as nitrogen and phosphorus to plants (Abdenaceur et al. 2022 ; Li et al. 2015 ). Furthermore, Trichoderma forms symbiotic relationships with plant roots, improving the efficiency of water and nutrient absorption by plants (Mehetre et al. 2015). They also improve the diversity of soil microbial communities, enhancing the ecological purification capabilities of soil (Mao and Jiang 2021 ). These functions of Trichoderma help maintain the balance of the soil ecosystem and improve its sustainable production capacity (Woo et al. 2023 ; Pinto et al. 2024 ). Studies had shown that the application of Trichoderma biofertilizers to the rhizosphere of corn could effectively increase the content of organic matter, nitrogen, phosphorus, and potassium in the soil. After spraying the suspension of Trichoderma atroviride on corn at 15 and 25 days after germination, the soil ammonium nitrogen content increased by 15% (Fu et al. 2019 ). Liu et al. ( 2020 ) found that after applying a microbial fertilizer containing Trichoderma guizhouense during the growth process of chili peppers, the soil urease activity significantly increased, and the transformation of soil organic nitrogen accelerated, increasing the available nitrogen content in the rhizosphere soil of chili peppers, promoting their nutrient absorption, and thus increasing the yield of chili peppers. Pecan was primarily found in the Tianmu Mountain area, located at the border of Zhejiang and Anhui provinces. Thriving in the understory of mountain slopes or valleys that were rich in humus, it was an economically important forest tree in China (Guo et al. 2004 ). In recent years, the excessive use of chemical fertilizers and pesticides, coupled with the long-term overexploitation of pecan forests, led to soil fertility degradation, frequent soil-borne diseases, and reduced yields, causing economic losses for farmers (Fang et al. 2020 ). There was an urgent need for a biofertilizer product that could improve the fertility of pecan forest land and was also friendly to the micro-ecological environment. Trichoderma spp., as a green biological pesticide, had potential in environmental remediation and plant health, but their application in the pecan industry had been relatively understudied. This study explored the effects of the Trichoderma asperellum TCS007 (isolated from Antarctic marine sediments) on the physiological and biochemical indicators of pecan plants, the physicochemical properties of rhizosphere soil, and soil enzymatic activity. High-throughput sequencing was used to analyze the diversity and structural composition of the microbial community in the rhizosphere soil of pecan, providing a theoretical basis for the development of TCS007 as a biofertilizer product, thereby contributing to the stable development of the pecan industry. Materials and Methods Experimental materials and equipment Tested microbial strains and plants The T. asperellum TCS007 was isolated from Antarctic marine sediments by our laboratory. The strain was preserved at the China General Microbiological Culture Collection Center (CGMCC), with the accession number CGMCC No.15677. One-year-old pecan seedlings, provided by the Panmugang Modern Forestry Demonstration Base, located in Lin'an District, Hangzhou City, Zhejiang Province, were transplanted into plastic pots filled with a mixture of peat soil/vermiculite/sand (2:1:1, v/v/v) as the growth substrate. After transplanting, the seedlings were placed in a greenhouse under 12 h photoperiod at 26°C and allowed to acclimatize for 30 days before use. Reagents and Culture Media Plant hormone indole-3-acetic acid (IAA) and gibberellin (GA) assay kits were purchased from Nanjing Camilo Bioengineering Co., Ltd.; enzyme activity assay kits for soil catalase, urease, and β-glucosidase were purchased from Beijing Boxbio Science & Technology Co., Ltd., and soil genomic DNA extraction kits were purchased from Tiangen Biotech Co., Ltd. PDA and PDB media (Li and He 2005 ) were used for cultivating the TCS007. Tryptone Beef Broth, Martin medium, and Modified Gause's No. 1 medium (Li et al. 1996 ) were used for the cultivation of bacteria, fungi, and actinomycetes from the soil samples. Experimental Methods Preparation of TCS007 spore suspension and liquid fermentation crude extract solution For spore suspension preparation; two agar plugs were punched from the edge of a revived TCS007 colony on PDA using a 9 mm diameter borer and inoculated into 100 mL PDB medium in a 250 mL flask for cultivation at 28°C at 180 rpm for 7 days. The liquid culture was then filtered through sterile gauze three times to remove the mycelia and debrises, obtained the fermentation filtrate of TCS007. The concentration of the spore suspension was determined using a hemocytometer and adjusted to 1.0 × 10 6 and 1.0 × 10 8 spores/mL for subsequent use. Preparation of TCS007 liquid fermentation crude extract solution: The TCS007 fermentation filtrate was acquired as aforementioned. According to the method described by Liu et al. ( 2019 ), the fermentation filtrate of TCS007 was extracted with an equal volume of ethyl acetate three times. The upper organic phase was collected and dried using a saturated sodium chloride solution and anhydrous sodium sulfate. After that, the extract was concentrated under reduced pressure using a rotary evaporator, redissolved in acetone, and collected in a centrifuge tube. Once the solvent had evaporated, the TCS007 organic phase extract was obtained. It was then diluted with sterile water to a concentration of 50 mg/mL for subsequent use. Experimental Design and Seedlings Inoculation with T. asperellum TCS007 The pot experiment was conducted using a single-factor completely randomized block design with a total of five treatments (Table 1 ). Each treatment consisted of 20 seedlings as replicates, and the treatments were initiated subsequent to the seedlings that had been acclimatized over a period of 30 days. Each treatment was applied three times, with a 30-day interval between successive applications. Table 1 Treatments used in this study Treatment Test agent Treatment concentration Treatment method A sterile water - Root irrigation was performed with a volume of 50 mL B sterilized PDB - C TCS007 spore suspension 1.0 × 10 8 spores/mL D TCS007 spore suspension 1.0 × 10 6 spores/mL E TCS007 liquid fermentation crude extract 50 mg/mL Determination of Physiological and Biochemical Indices of Seedlings Three months after treatment, ten seedlings were randomly selected from each treatment for further analysis. Plant height and ground diameter were measured using a millimeter scale ruler. Chlorophyll content was determined by UV-2801 spectrophotometry (Hitachi, Ltd., Japan.) (Pan et al. 2017). Leaf IAA and GA content were quantified using enzyme-linked immunosorbent assay kits, employing a double-antibody sandwich method (Chen et al. 2011 ). The activities of Superoxide dismutase (SOD) and Peroxidase (POD) enzymes were determined using the nitroblue tetrazolium method and the guaiacol method, respectively (Shahzad et al. 2018 ). With each treatment group replicated ten times. Determination of soil physicochemical properties, enzyme activities, and microbial counts During the leaf expansion stage (mid-April), the new shoot growth stage (mid-July), and the defoliation stage (early October) of the pecan seedlings, rhizosphere soil samples from each treatment group were collected. The 20 pecan seedlings in a treatment were randomly divided into 5 groups. For each seedling, 12.5 g of soil sample was taken from the region rich in fine roots approximately 1 mm in diameter in the soil layer at a depth of 10–30 cm (Zhang et al. 2021 ). The soil samples taken from the four seedlings assigned in the same group in one treatment were mixed thoroughly to form a single soil sample. In total, five soil samples as five replicates were collected for each treatment for subsequent processing. The soil samples were air-dried at 25°C, passed through a 1 mm sieve, collected in sterile sample bags, and then stored at 4°C for future use. The content of organic matter was determined using the dichromate volumetric method (Zhang and Fen 2023). The ammonium nitrogen content was determined using the titration method (Li et al. 2021b ). The readily available phosphorus content was determined using the extraction-antimony molybdate colorimetric method (Li et al. 2021a ). The content of ammonium nitrogen was determined using the titration method (Cheng, 2014 ). The available potassium content was determined using the atomic absorption spectrophotometer with a flame photometer method (Guo et al. 2012 ). The soil pH value was determined using the electrode method (Fu et al. 2012 ). With each treatment group replicated five times. Enzyme activities of soil catalase, urease, and β-glucosidase were measured using assay kits from Beijing Boxbio Science & Technology Co., Ltd., following the manufacturer’s instructions. Catalase activity was assessed by monitoring the decomposition of H₂O₂, which reduced absorbance at 240 nm. One unit of catalase activity was defined as the amount of enzyme degrading 1 mmol H₂O₂ per gram of air-dried soil. Urease activity was determined by the formation of indophenol blue (absorbance at 630 nm) from NH₃-N generated by urea hydrolysis. One unit of urease activity corresponded to the production of 1 µg NH₃-N per gram of soil. β-Glucosidase activity was measured by the hydrolysis of p-nitrophenyl-β-D-glucopyranoside to p-nitrophenol, with absorbance at 400 nm. One unit of β-glucosidase activity was defined as the amount of enzyme generating 1 µmol p-nitrophenol per gram of soil. With each treatment group replicated five times. The numbers of bacteria, fungi, and actinomycetes in the soil samples from each treatment group were determined using the dilution plating method. Briefly, in a 250 mL Erlenmeyer flask, 50 mL of sterile water was added. Then, 5 g of fresh soil sample was introduced into the flask and agitated at room temperature for 10 min. Subsequently, 1 mL of the soil suspension was transferred to 9 mL of sterile water, and the resulting suspension was serially diluted by a factor of 10 up to 10 − 6 . Based on preliminary experiments, for this trial, fungi were diluted between 10 − 3 and 10 − 1 , while bacteria and actinomycetes were diluted between 10 − 5 and 10 − 3 . An aliquot of 50 µL of the diluted solution was spread evenly onto the culture medium with each dilution repeated three times (Li and He 2005 ). Tryptone Soya Broth agar, Martin medium, and Modified Gause's No. 1 medium (Li et al. 1996 ) were utilized for the isolation and enumeration of bacteria, fungi, and actinomycetes, respectively. With each treatment group replicated five times. Construction of Internal Transcribed Spacer (ITS) and 16S rRNA Gene Libraries and Subsequent High-Throughput Sequencing Analysis During the leaf expansion stage of the pecan seedlings, soil microbial DNA was extracted from the five soil samples in each treatment group. The quality of the extracted DNA was assessed using 1.5% agarose gel electrophoresis and a spectrophotometer. The V3-V4 region of the bacterial 16S rRNA gene and the ITS1 region of the fungal ITS sequence were amplified using the universal primers 338F (5'-ACTCCTACGGGAGGCAGCAG-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3'), as well as ITS1F (5'-CTTGGTCATTTAGAGGAAGTAA-3') and ITS2R (5'-GCTGCGTTCTTCATCGATGC-3'). PCR amplification was conducted in a 20 µL reaction system using TransStart FastPfu DNA Polymerase and TaKaRa rTaq DNA Polymerase. After amplification, the PCR products were recovered through purification, detection, and quantification steps. Subsequently, the amplified products were mixed in equimolar concentrations to form a single sequencing library. The constructed library was quality checked, and qualified libraries were sequenced using the Illumina MiSeq sequencing platform (PE300). FLASH v1.2.7 software was used to assemble reads from each sample based on overlaps, yielding raw Tag data (Magoč et al. 2011). Trimmomatic v0.33 software was employed to filter the raw Tag data, resulting in high-quality Tag data (Bolger et al. 2014 ). UCHIME v4.2 software was utilized to identify and remove chimeric sequences, obtaining the final data. For soil microbial genomic data, Operational Taxonomic Units (OTUs) clustering analysis was conducted using USEARCH software. Biostatistical analysis on OTUs at 97% similarity level was performed (Edgar et al. 2013). The clustered OTUs were then used for various analyses: alpha diversity analysis (ACE index, Chao1 index, Shannon index, Simpson index, and Coverage index), beta diversity analysis, and species composition and difference analysis. Alpha diversity indices were calculated under different random samplings using Mothur software, while beta diversity analysis was conducted with QIIME software. Non-metric multidimensional scaling (NMDS) analysis and heatmap analysis were executed using R software, and the ANOSIM test was applied for statistical comparisons. All tables were prepared with Excel 2021 software, and the analyses were performed on the Majorbio Cloud Platform ( www.majorbio.com ) in Shanghai. Statistical Analysis and Data Interpretation Physiological and biochemical data of plants and soil were statistically analyzed using Excel 2021 software. To determine differences in physiological and biochemical indices of pecan seedlings, soil physicochemical properties, and soil enzymatic activities, one-way analysis of variance (ANOVA) and Duncan's test within IBM SPSS 22.0 software (IBM Corporation, New York, NY, USA) were employed. Results The Impact of TCS007 on the Physiological and Biochemical Indicators of Pecan Plant Plant height, chlorophyll content, and ground diameter were measured for each treatment group. Compared to the control group A, treatment group C showed a height increase of 31.8%, a chlorophyll content increase of 225.8%, and a ground diameter increase of 329.6%. Treatment group E exhibited increases of 19.3%, 56.3%, and 29.2%, respectively, in height, chlorophyll content, and groun d diameter, while treatment group B showed no significant differences in all three indicators. The experimental results indicated that both the TCS007 spore suspension and crude extract promoted the growth of pecan plants, with the optimal growth-promoting effect observed at a concentration of 1.0 × 10 8 spores/mL for the TCS007 spore suspension (Table 2 ). Table 2 The impact of Trichoderma inoculation on seedling biomass Treatment groups Plant Height (cm) Ground diameter (cm) Chlorophyll content (mg·g -1 ) A 6.99 ± 0.19 c 0.27 ± 0.36 b 6.28 ± 0.19 d B 7.28 ± 0.24 c 0.35 ± 0.82 b 6.94 ± 0.07 d C 9.21 ± 0.23 a 1.16 ± 0.84 a 20.46 ± 0.96 a D 8.71 ± 0.40 ab 1.02 ± 0.89 a 17.38 ± 1.03 b E 8.34 ± 0.29 b 0.35 ± 0.40 e 9.82 ± 0.64 c Note: Different letters in the columns indicate significant differences ( p < 0.05), according to Duncan’s new multiple range test. In terms of phytohormone content, the C treatment group showed an increase of 60.0% in IAA and 60.8% in GA in the leaf tissues, compared to the A treatment group. The E treatment group exhibited increases of 26.2% in IAA and 31.4% in GA, respectively (Fig. 1 a-b). Regarding defense enzymes, the C treatment group had a 53.5% increase in superoxide dismutase (SOD) and a 140.0% increase in peroxidase (POD) enzyme activities in the leaf tissues. The treatment of group D showed even higher increases by 69.5% and 166.1% for SOD and POD, respectively (Fig. 1 c-d). The B treatment group did not exhibit significant differences in either phytohormone content or defense enzyme activities. The experimental results suggested that both the TCS007 spore suspension and crude extract could enhance hormone content and defense enzyme activities in the leaf tissues of pecan plants, with the most pronounced effects observed at a TCS007 spore suspension concentration of 1.0 × 10 8 spores/mL. The Impact of TCS007 on the Physicochemical Properties of Soil in the Rhizosphere of pecan During the leaf expansion, shoot growth, and defoliation stages of pecan seedlings, the organic matter content in the treatment of group C increased by 19.4%, 17.0%, and 10.9%, respectively, the ammonium nitrogen content increased by 20.0%, 21.3%, and 10.3%, respectively, and the available phosphorus content increased by 23.7%, 16.7%, and 17.7%, respectively. The soil pH values were elevated to 7.53, 6.96, and 6.76, respectively. In the treatment of group D, the organic matter content increased by 12.7%, 13.2%, and 6.3%, the ammonium nitrogen content increased by 17.3%, 15.3%, and 8.0%, and the available phosphorus content increased by 13.7%, 14.4%, and 15.2%, with soil pH values elevated to 7.32, 6.84, and 6.62, respectively. However, there were no significant differences in available potassium (Table 3 ). The experimental results indicated that the TCS007 spore suspension significantly increased the organic matter content, ammonium nitrogen content, available phosphorus content, and soil pH in the rhizosphere soil of pecan, while the crude extract treatment group showed no significant differences. Table 3 The influence of different treatments on the physicochemical properties of the rhizosphere soil during various growth stages of pecan Pecan growth stages Treatment groups Organic matter content/% Ammonium Nitrogen /mg·kg -1 Available Phosphorus /mg·k -1 Available Potassium /mg·kg -1 pH value Leaf expansion stage A 2.48 ± 0.73 c 44.57 ± 2.38 b 22.92 ± 2.19 c 54.63 ± 3.25 a 6.32 ± 5.36 b B 2.63 ± 1.83 b 46.76 ± 2.03 b 23.33 ± 0.97 c 53.96 ± 1.64 a 6.35 ± 0.89 b C 2.96 ± 0.82 a 53.49 ± 1.97 a 28.36 ± 1.87 a 54.38 ± 2.33 a 7.53 ± 3.28 a D 2.84 ± 0.62 a 52.28 ± 1.23 a 26.06 ± 0.93 b 54.20 ± 1.47 a 7.32 ± 2.08 a E 2.52 ± 1.32 c 43.66 ± 1.28 b 22.67 ± 1.65 c 54.32 ± 1.48 a 6.49 ± 1.38 b Shoot growth stage A 2.42 ± 1.48 b 45.23 ± 0.77 b 23.64 ± 1.93 b 55.49 ± 2.39 a 6.44 ± 2.01 b B 2.45 ± 0.56 b 46.18 ± 1.63 b 24.11 ± 0.78 b 55.29 ± 1.47 a 6.37 ± 1.84 b C 2.83 ± 1.04 a 54.88 ± 2.31 a 27.58 ± 1.27 a 54.18 ± 0.48 a 6.96 ± 0.49 a D 2.74 ± 1.23 a 52.16 ± 1.71 a 27.05 ± 0.85 a 53.26 ± 1.39 a 6.84 ± 1.63 a E 2.43 ± 0.73 b 45.87 ± 1.02 b 23.17 ± 1.38 b 52.93 ± 3.19 a 6.53 ± 0.76 b Defoliation stage A 2.39 ± 1.76 b 42.33 ± 1.74 b 21.95 ± 1.47 b 50.88 ± 2.38 a 6.27 ± 1.68 b B 2.41 ± 1.27 b 42.89 ± 2.18 b 22.16 ± 1.29 b 48.87 ± 0.38 a 6.33 ± 3.94 b C 2.65 ± 0.93 a 46.71 ± 0.87 a 25.83 ± 0.48 a 49.73 ± 1.20 a 6.76 ± 2.06 a D 2.54 ± 1.32 a 45.71 ± 0.97 a 25.29 ± 1.94 a 49.15 ± 0.68 a 6.62 ± 1.83 a E 2.31 ± 1.76 b 41.97 ± 1.22 b 21.37 ± 0.99 b 51.28 ± 0.77 a 6.19 ± 6.93 b Note: Different letters in the columns indicate significant differences ( p < 0.05), according to Duncan’s new multiple range test. The Impact of TCS007 on the Enzymatic Activity of Rhizosphere Soil in Pecan During the leaf expansion stage of the pecan, the activities of soil catalase, urease, and β-glucosidase in the C treatment group significantly increased by 55.2%, 78.4%, and 62.0%, respectively, compared to the A treatment group. The E treatment group also showed increases in soil enzyme activities of 32.8%, 22.4%, and 96.3%. During the shoot growth stage, the C treatment group showed an increase in soil enzyme activities of 46.1%, 37.6%, and 44.1%, while the E treatment group's increases were 36.5%, 9.0%, and 65.8%. By the time of defoliation stage, the C treatment group's soil enzyme activities had further increased by 30.5%, 20.8%, and 23.2%, the E treatment group's activities increased the soil enzymatic activity by 25.5%, 15.9%, and 22.7%, and the B treatment group showed no significant difference. The experimental results indicated that the spore suspension and crude extract of TCS007 could effectively enhance soil enzyme activity, with the crude extract showing the most pronounced effect (Fig. 2 ). The Influence of TCS007 on the Culturable Microbial Population in the Rhizosphere Soil of Pecan During the leaf expansion stage, compared to the A treatment group, the microbial population in the rhizosphere soil of the C treatment group significantly increased, while in the E treatment group, the number of fungi decreased by 69.2%, bacteria by 57.8%, and actinomycetes by 66.7%. During the shoot growth stage, compared to the A treatment group, the microbial population in the rhizosphere soil of the C treatment group significantly increased again, with a 49.6% reduction in fungal numbers, a 21.1% increase in bacterial numbers, and a significant 79.3% decrease in actinomycetes in the E treatment group. In the defoliation stage, compared to the A treatment group, the number of fungi and bacteria in the rhizosphere soil of the C treatment group significantly increased, while the actinomycetes decreased by 7.5%. In the E treatment group, the numbers of fungi and actinomycetes decreased by 39.2% and 16.4%. Respectively. In the B treatment group, the numbers of fungi and bacteria only significantly increased during the leaf expansion stage, with no significant changes in the microbial population in the rhizosphere soil during other stages. The experimental results indicated that during the growth process of pecan plants, the number of fungi in the rhizosphere soil first decreased and then increased after treatment with TCS007 spore suspension (Fig. 3 a), bacteria decreased and then stabilized (Fig. 3 b), and actinomycetes significantly decreased (Fig. 3 c). After treatment with TCS007 crude extract, the number of fungi in the rhizosphere soil significantly decreased (Fig. 3 a), the number of bacteria significantly increased after the leaf expansion stage (Fig. 3 b), and the number of actinomycetes did not significantly change (Fig. 3 c). The spore suspension and crude extract of TCS007 both significantly affected the number of soil microorganisms, with the spore suspension of TCS007 having the most pronounced effect (Fig. 3 ). The Influence of TCS007 on the Microbial Composition of the Rhizosphere Soil in Pecan Microbial Sequencing Data Analysis The statistics of the sequencing data after quality control, filtering, and assembly for the five treatment groups are presented in Table 4 . The effective bacterial sequences were 28089, 21237, 26931, 25779, and 24914; and the effective fungal sequences were 49472, 49101, 45903, 39691, and 42696. The sequencing quality, measured by the Q30 score, ranged from 97–99%, while the Q20 score ranged from 99–99.5% (Table 4 ). Table 4 Summary of basic sequencing data for each sample Microorganisms Treatment Shortest tags Longest tags Mean tags Effective tags Total bases Bacteria A 301 504 419 28089 21438243 B 317 527 417 21237 21312593 C 317 497 418 26931 21395076 D 319 456 419 25779 19405297 E 270 469 418 24914 19781631 Fungi A 141 526 237 49472 12587811 B 141 533 232 49101 12441487 C 142 384 245 45903 12569760 D 143 522 237 39691 10209814 E 142 514 236 42696 11074245 Microbial Community Diversity Analysis Alpha diversity indices analysis revealed that the coverage indices for all treatment groups were close to 1, indicating that the sequencing results could accurately reflect the actual conditions of the soil samples tested. The ACE and Chao indices were indicative of the richness of soil microbial communities, while the Simpson and Shannon indices reflected the diversity of these communities. Among bacterial samples, the analysis of Chao1 and ACE indices across treatment groups revealed that the bacterial community richness in treatment groups C and D was significantly higher than in groups A and B ( p < 0.05), with the bacterial community richness in pecan rhizosphere soil increasing as the concentration of TCS007 spores increased. Analysis of the Simpson and Shannon indices showed that the bacterial community diversity in treatment group C was significantly higher than in other groups ( p < 0.05), with no significant difference in diversity between groups B and D compared to group A. After treatment with group E, the bacterial community richness and diversity in the pecan rhizosphere were significantly reduced compared to other treatment groups ( p < 0.05). In the case of fungal samples, the analysis of Chao1 and ACE indices demonstrated that the richness of fungal communities in treatment groups C and D was significantly higher than in other groups ( p < 0.05). Analysis of the Shannon and Simpson indices indicated that the fungal community diversity in groups C and D was significantly higher than in group A ( p < 0.05). However, the richness and diversity of fungal communities in group E were significantly lower than in other treatment groups ( p < 0.05) (Table 5 ). Table 5 The impact of various treatments on the diversity indices of the microbial community Microorganisms Treatment Shannon Index Simpson Index Chao Index ACE Index Coverage % Bacteria A 6.79 ± 0.02a 0.23 ± 0.01b 1579.79 ± 0.40b 1591.28 ± 0.34c 0.99 ± 0.01a B 6.79 ± 0.01a 0.24 ± 0.15b 1634.00 ± 0.14b 1666.65 ± 0.36b 0.99 ± 0.01a C 6.83 ± 0.02a 0.59 ± 0.03a 1708.34 ± 0.36a 1725.41 ± 0.56a 1.00 ± 0.00a D 6.80 ± 0.11a 0.24 ± 0.01b 1699.44 ± 0.28a 1714.37 ± 0.19a 0.99 ± 0.01a E 6.49 ± 0.19b 0.20 ± 0.01c 1315.56 ± 0.60c 1337.70 ± 0.20d 0.99 ± 0.01a Fungi A 4.35 ± 0.01c 0.07 ± 0.01c 515.00 ± 0.48d 515.00 ± 0.18b 1.00 ± 0.00a B 4.42 ± 0.02b 0.53 ± 0.03a 551.67 ± 0.44c 553.87 ± 0.43b 1.00 ± 0.00a C 4.75 ± 0.05a 0.12 ± 0.01b 630.91 ± 0.08a 632.87 ± 0.19a 1.00 ± 0.00a D 4.73 ± 0.03a 0.12 ± 0.01b 568.41 ± 0.18b 569.46 ± 0.57a 1.00 ± 0.00a E 3.14 ± 0.03d 0.05 ± 0.02c 414.17e 416.38 ± 0.35c 1.00 ± 0.00a Note: Different letters in the columns indicate significant differences ( p < 0.05), according to Duncan’s new multiple range test. Microbial Community Compositional Differences Analysis Beta diversity analysis revealed that compared to the treatment group A, the soil bacterial community compositions of the other treatment groups showed significant differences, with the most pronounced difference observed between treatment group C and group A, while treatment groups D and E exhibited a higher similarity in their bacterial community compositions (Fig. 4 a). In comparison to treatment group A, the soil fungal community compositions of treatment groups C and D were significantly different, whereas the fungal community composition of treatment group E showed a higher similarity to that of treatment group A (Fig. 4 b). The analysis of soil microbial community compositional differences revealed that the structure of the community in different samples was known at various taxonomic levels, such as domain, kingdom, phylum, class, order, family, genus, and species. The bacterial community in the rhizosphere soil of pecan was primarily composed of the phyla Actinobacteria , Proteobacteria , Acidobacteria , and Chloroflexi , with Actinobacteria , Proteobacteria , and Acidobacteria being the dominant. Compared to treatment A, treatment C increased the abundance of Actinobacteria by 36.19% and significantly increased the abundance of Firmicutes by 301.57%. In contrast, the abundance of Acidobacteria and Chloroflexi in the rhizosphere soil bacterial community decreased by 45.49% and 34.92%, respectively (Fig. 5 a). Additionally, treatment C reduced the abundance of the Acidobacteriaceae and RB41 genera in the bacterial community by 46.25% and 61.03%, respectively, while the abundance of the genera Nocardioides , Bacillus , Micrococcaceae , and Agromyces increased, with Bacillus showing a significant increase of 377.65%. However, treatments B, D, and E did not show significant differences in the abundance of the bacterial community in the rhizosphere soil compared to treatment A (Fig. 5 b, Fig. 6 ). The fungal community in the rhizosphere soil of pecan was predominantly composed of the phyla Ascomycota , Mortierellomycota , and Basidiomycota , with Ascomycota and Mortierellomycota being the dominant. Compared to the A treatment group, the abundance of Ascomycota in the C treatment group increased by 23.72%, while the abundance of Mortierellomycota and Basidiomycota decreased by 41.18% and 21.87%, respectively (Fig. 7 a). Furthermore, the abundance of the genera Trichoderma and Apiotrichum in the C treatment group significantly increased, while other fungi were generally inhibited. Notably, the genera Mortierella , Neocosmospora , Fusarium , Chaetomium , Talaromyces , Aspergillus , Cladosporium , and Acremonium showed the most pronounced inhibition. Specifically, the abundance of Mortierella , Neocosmospora , Fusarium , and Cladosporium decreased by 41.10%, 45.77%, 55.71%, and 46.15%, respectively. However, the B, D, and E treatment groups showed no significant differences (Figs. 7 b and 8 ). Discussion As a commonly used biocontrol agent, Trichoderma species exhibit a range of efficacy. Upon application, Trichoderma played a crucial role in the rhizosphere by competing against other microbes, particularly plant pathogens (Ferreira and Musumeci 2021 ). This competition was multifaceted and included direct antagonism, nutrient and space competition, and the production of antifungal compounds (Manzar et al 2022 ). Additionally, Trichoderma could outcompete pathogens for nutrients and space by rapidly colonizing plant roots and the surrounding soil (Tyśkiewicz et al. 2022 ). This competition not only suppressed the growth of pathogens but also promoted plant health by enhancing nutrient uptake and inducing systemic resistance in plants.(Dutta et al. 2022 ). Then, it produces plant growth hormones that enhance the solubility of nutrients in the soil, improves rhizosphere microecology, and thereby facilitates nutrient uptake by plants, promoting growth and increasing yield (Contreras-Cornejo et al. 2024 ; Li et al. 2024 ; Wei et al. 2024). Additionally, Trichoderma induces the production of defense enzymes in plants, such as peroxidase (POD) and superoxide dismutase (SOD), which play a crucial role in the plant resistance to stress and diseases (Ahmad et al. 2015 ; Zhang et al. 2016 ; Pacheco-Trejo et al. 2022 ). Trichoderma elicits plant immune responses through multiple mechanisms. It secretes microbe-associated molecular patterns (MAMPs) recognized by plant cell surface pattern recognition receptors (PRRs), triggering basal immunity (Alfiky and Weisskopf 2021 ). Our study revealed that the TCS007 spore suspension and crude extracts significantly promoted the growth of pecan seedlings and enhanced chlorophyll content in leaves. Concurrently, there was a notable increase in the levels of plant hormones (IAA, GA) and the activity of defense enzymes (POD, SOD) in the leaves, with the optimal effect observed at a spore suspension concentration of 1×10 8 spores/mL. Studies showed that Trichoderma strains isolated from various regions could secrete IAA, enhancing the photosynthetic rate and promoting the growth of plants such as cucumber and corn (Vinale et al. 2008a ; Vargas et al. 2009 ). Zhang et al. ( 2021 ) found that Trichoderma harzianum significantly increased the plant height, basal diameter, and fresh weight of Malus hupehensis Rehd. Moreover, Trichoderma species were capable of producing a variety of secondary metabolites, including 6-pentyl-2H-pyran-2-one (6PP), Koninginin A5, gliotoxin, and viridin, which played significant signaling roles in plant-microbe interactions (Forde et al. 2014; Garnica-Vergara et al. 2015 ; Moisan et al. 2021 ). It was demonstrated that Trichoderma atroviride could produce at least 25 types of volatile organic compounds, encompassing alcohols, ketones, alkanes, ethylene, monoterpenes, and sesquiterpenes, among which gaseous ethylene acted as a core signaling molecule in Trichoderma -plant interactions (Lee et al. 2016 ; Estrada-Rivera et al. 2019 ). Vinale et al. ( 2008b ) observed growth promotion in etiolated pea stems treated with the main secondary metabolites produced by different Trichoderma strains, harzianic acid, and 6PP. Soil nutrients, often existing in sparingly soluble or insoluble forms, impeded nutrient cycling, highlighting the importance of enhancing nutrient availability. Trichoderma species, through the secretion of organic acids, activated soil nutrients and promoted plant absorption, playing a crucial role in rehabilitating degraded soils (Sindhu et al. 2022 ; Singh et al. 2024 ). Our study demonstrated that TCS007 spore suspension significantly increased organic matter, ammonium nitrogen, available phosphorus content, and pH value in the rhizosphere soil of pecan, with the optimal effect at a concentration of 1×10 8 spores/mL. Asghar and Kataoka. (2021) found that soil phosphatase activity had increased by more than 10% after inoculation with Trichoderma RW309 for two months, accelerating the transformation of organic phosphorus compounds or inorganic phosphates in the soil. The physiological metabolism of different microbial communities was closely related to the production of soil enzymes, which could reflect the presence and activity of corresponding functional microbes (Mao and Jiang. 2021; Jia et al. 2024 ; Aljeddani et al. 2024 ). Trichoderma species rapid colonization capability enhanced rhizosphere-soil contact, promoting the secretion of extracellular enzymes such as catalase, urease, and β-glucosidase, as well as organic acids (Solomon et al. 2024 ). Catalase served as an indicator of soil microecology, urease reflected the soil's nitrogen supply capacity, and β-glucosidase provided a carbon source for soil microbes (Woese et al. 1987; Stott et al. 2010 ; Su et al. 2016 ). Our results indicated that TCS007 spore suspension and crude extracts effectively enhanced soil enzyme activity, with the most significant effect observed at a concentration of 50 mg/L for the crude extracts. In a cabbage pot experiment, the treatment with Trichoderma increased soil transformation enzyme activity by 45%, effectively promoting nutrient transformation and improving the soil ecological environment (Shi et al. 2021 ). Forest soil microbes played a pivotal role in ecosystems, encompassing soil formation, development, and the maintenance of ecological balance. The ratio of soil bacteria, fungi, and actinomycetes served as a vital indicator of soil fertility, with their quantities showing a significant positive correlation with soil nutrients and crop yields (Hang et al. 2022 ). Increasing the populations of rhizosphere bacteria and actinomycetes aided in enhancing soil nitrogen and phosphorus content, promoting plant root development and nutrient absorption (Mäkipää et al. 2017 ; Xiong et al. 2017 ; Harsonowati et al. 2020 ; Chen et al. 2021 ). Soil microbial diversity and richness are considered critical for the integrity, function, and long-term sustainability of soil ecosystems. The latter are usually reduced by agricultural perturbations (Fu et al. 2019 ). Our study found that TCS007 spore suspension significantly increased the number of culturable bacteria and actinomycetes in the rhizosphere soil of pecan during the leaf expansion and shoot growth stage, particularly the beneficial microbes Nocardia and Bacillus , enhanced the abundance of bacterial community richness and bacterial community diversity in the soil. Concurrently, TCS007 spore suspension increased the abundance of Trichoderma and Apiotrichum fungi, suppressing the growth of plant pathogens such as Fusarium , Colletotrichum , and Magnaporthe oryzae , thus promoting the growth of beneficial fungal communities and inhibiting pathogenic fungi. Consequently treating soil with TCS007 spore suspension would altering the uniformity and richness of soil species, and altering the soil’s microbial diversity. Fu et al. ( 2019 ) study indicated that Trichoderma treatment impacts bacterial diversity in the rhizosphere soil of maize. The diversity of the bacterial community in soil following Trichoderma treatment was lower than that in the control, untreated, soil. This might be associated with the fact that Trichoderma promotes plant growth and plant nutrient absorption from the soil. Hou et al. ( 2021 ) demonstrated that Trichoderma harzianum TH62 effectively inhibited soil-borne pathogens such as Fusarium oxysporum and Alternaria alternata . In addition, T. harzianum ESALQ-1306 and T. asperellum BRM-29104 had a minor impact on soil fungal abundance, they shifted the soil microbial community from fungi to bacteria, maintaining microbial balance and reducing crop diseases and pests (Cordier and Alabouvette. 2009; Zhang et al. 2019 ; Silva et al. 2021 ). These results indicated that TCS007 spore suspension could regulate soil microbial communities, enhancing soil fertility and crop health. Future research should focus on elucidating the specific mechanisms underlying the beneficial effects of T. asperellum TCS007 on pecan plants and their rhizosphere environment. Long-term field trials are needed to assess the sustained impact of TCS007 on soil health and plant growth under varying environmental conditions. Additionally, studies should explore the potential synergistic effects of TCS007 with other soil amendments or agricultural practices. Investigating the genetic and biochemical pathways involved in the interactions between TCS007 and pecan plants could provide valuable insights into optimizing its application as a biological fungal fertilizer. Conclusions In a pioneering effort, our research team has introduced the marine-derived strain T. asperellum TCS007 to the economically important tree species pecan. This study demonstrated that treatment with TCS007 spore suspension or crude extract promoted the growth of pecan seedlings, with significant increases in plant height, ground diameter, and chlorophyll content compared to the CK. Additionally, the contents of IAA and GA in the leaves, as well as the activities of SOD and POD enzymes, were significantly enhanced. The best growth-promoting effect was observed when the TCS007 spore suspension was applied at a concentration of 1.0 × 10 8 spores/mL. In addition, the levels of organic matter, ammonium nitrogen, and available phosphorus content in the rhizosphere soil, as well as the activities of soil catalase, urease, and β-glucosidase, were significantly higher in the treatment groups than those in the CK, and the soil pH shifted from slightly acidic to slightly alkaline after treatment. The soil microbial communities of pecan seedlings treated with TCS007 were analyzed during the leaf expansion stage, high-throughput sequencing revealed that the abundance of beneficial microbes, such as Nocardia , Bacillus , Micrococcus , and Mycobacterium in the bacterial community and Trichoderma and Penicillium in the fungal community, significantly increased, while the abundance of Acidobacterium and RB41 in bacterial community and other fungal genera decreased in the group treated with TCS007 spore suspension. In summary, the TCS007 spore suspension could effectively promote the growth and development of pecan plants, improved the stability of the rhizosphere soil microecology of pecan, and increased the content of soil nutrients. It had a positive effect on the prevention and control of soil-borne diseases in pecan and the regulation of soil pH. These results indicated that T. asperellum TCS007 had great potential in being developed into a biological fungal fertilizer product for pecan. Declarations Declarations The authors have no relevant financial or non-financial interests to disclose. Founding This study was supported by the Science and Technology Collaboration Program of Zhejiang Province (2023SNJF035), the National Key Research and Development Program of China (2022YFD1700400). References Abdenaceur, R., Farida, Bt., Mourad, D., Rima, H., Zahia, O., Fatma, S. Effective biofertilizer Trichoderma spp. isolates with enzymatic activity and metabolites enhancing plant growth. 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Soil Science Society of America Journal. 2010, 74, 107-119. https://doi.org/10.2136/sssaj2009.0029. Su, C.L.; Wang, H.W.; Xie, X.G.; Zhang, W.; Li, X.G.; Wang, X.X.; Dai, C.C. Effects of endophytic fungi and Atractylodes lancea powder on rhizosphere microflora and trace elements during continuous peanut cropping. ACAT Ecologica Sinica. 2016, 36(7), 2052-2065. DOI: 10.5846/stxb201409171842. Tyśkiewicz, R.; Nowak, A.; Ozimek, E.; Jaroszuk-Ściseł, J. Trichoderma : The Current Status of Its Application in Agriculture for the Biocontrol of Fungal Phytopathogens and Stimulation of Plant Growth. Int. J. Mol. Sci. 2022, 23, 2329. https://doi.org/10.3390/ijms23042329. Vargas, W.A.; Mandawe, J.C.; Kenerley, C.M. Plant-derived sucrose is a key element in the symbiotic association between Trichoderma virens and maize plants. Plant physiology. 2009, 151(2), 792-808. https://doi.org/10.1104/pp.109.141291. 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Trichoderma : a multipurpose, plant-beneficial microorganism for eco-sustainable agriculture. Nature Reviews Microbiology. 2023, 21, 5, 312-326. Xiong, W.; Guo, S.; Jousset, A.; Zhao, Q.Y.; Wu, H.S.; Li, R.; Kowalchuk, G.A.; Shen, Q.R. Bio-fertilizer application induces soil suppressiveness against Fusarium wilt disease by reshaping the soil microbiome. Soil Biology and Biochemistry, 2017, 114: 238-247. https://doi.org/10.1016/j.soilbio.2017.07.016. Yang, Y.Y.; Zheng, K.B.; Chen, S.; Chen, J. Plant growth-promoting and stress tolerance inductive effects of Trichoderma asperellum TCS007 from marine habitat. Chinese Journal of Pesticide Science. 2023, 25(01), 132-139. doi: 10.16801/j.issn.1008-7303.2022.0147. Yao, X., Guo, H., Zhang, K., Zhao, M., Ruan, J., Chen, J. Trichoderma and its role in biological control of plant fungal and nematode disease. Frontiers in microbiology, 2023, 14, 1160551. https://doi.org/10.3389/fmicb.2023.1160551 Zhang, F.G.; Xu, X.X.; Huo, Y.Q.; Xiao, Y. Trichoderma -Inoculation and Mowing Synergistically Altered Soil Available Nutrients, Rhizosphere Chemical Compounds and Soil Microbial Community, Potentially Driving Alfalfa Growth. Front Microbiol. 2019, 9. https://doi.org/10.3389/fmicb.2018.03241. Zhang, R., Yan, Z., Wang, Y., Chen, X., Yin, C., Mao, Z. Effects of Trichoderma harzianum Fertilizer on the Soil Environment of Malus hupehensis Rehd. Seedlings under Replant Conditions. HortScience horts. 2021. 56(9), 1073-1079.https://doi.org/10.21273/HORTSCI15970-21. Zhang, S.B.; Feng, Z.Z. Study on determination of organic matter content in soil in some areas of Guangdong Province by potassium dichromate volumetric method. Guangdong Chemical Industry. 2023, 50(04), 189-191+213. Zhang, S.W., Gan, Y.T., Xu, B.L. Application of plant-growth-promoting fungi Trichoderma longibrachiatum T6 enhances tolerance of wheat to salt stress through improvement of antioxidative defense system and gene expression. Frontiers in plant science. 2016, 7, 1405. https://doi.org/10.3389/fpls.2016.01405. Zhang, Y. Z., Xu, J.; Nadia, R.; Wang, N. A Protocol for Citrus Rhizosphere and Rhizoplane Microbiome Sample Collection and Nucleic Acid Extraction. Bio-101 e2003680. 2021. Doi: 10.21769/BioProtoc.2003680. Zheng, K.B.; Lin, H.; Zhou, S.; Yuan, J.; Chen, J. Identification and antifungal activity of marine Trichoderma asperellum TCS007. Chinese Journal of Pesticide Science . 2020, 22(05), 801-807. doi: 10.16801/j.issn.1008-7303.2020.0075. Cite Share Download PDF Status: Published Journal Publication published 14 May, 2025 Read the published version in Plant and Soil → Version 1 posted Editorial decision: Accept 22 Apr, 2025 Reviewers agreed at journal 10 Apr, 2025 Reviewers invited by journal 10 Apr, 2025 Editor assigned by journal 08 Apr, 2025 First submitted to journal 07 Apr, 2025 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. 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-5756661","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":441055968,"identity":"cc8451e5-4636-48f2-b9cd-880c47d5991e","order_by":0,"name":"Hao Cao","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3ElEQVRIiWNgGAWjYDACCeaDDxIqJOoNGBgYgYwaYrSwJRt8OGOTANTCbPDgzDFitPCYSc5sSwNpYZN82MJMWAf/7B4zaZ4zh/PM2Q8fq0hsYGPgb+9OwG/JnWPF1jwVh4ste9LSbiTukGGQOHN2A14tBhLJG28DbWHccCDH7EbiGTagSC4hLQkG0rxtQC3n35gVJLYxE6MlxQjk/cQNN3LMGIjSInEjDRzIxgY3niVLJJw5xkPQL/wzksFRKWdwPvngxx8VNXL87b34tWAAHtKUj4JRMApGwSjACgAMPU+9ybv9xwAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0009-0004-9773-1180","institution":"Zhejiang Agriculture and Forestry University: Zhejiang A and F University","correspondingAuthor":true,"prefix":"","firstName":"Hao","middleName":"","lastName":"Cao","suffix":""},{"id":441055969,"identity":"09f056ca-9b10-4532-b64c-1ae4f8b27d7d","order_by":1,"name":"Xuesong Li","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Xuesong","middleName":"","lastName":"Li","suffix":""},{"id":441055970,"identity":"f1fd61fc-5fd8-40eb-b181-b15e8a56e332","order_by":2,"name":"Hao Han","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Hao","middleName":"","lastName":"Han","suffix":""},{"id":441055971,"identity":"34802664-47ed-4113-b449-1e9cdf2afaf3","order_by":3,"name":"Sai Chen","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Sai","middleName":"","lastName":"Chen","suffix":""},{"id":441055972,"identity":"33a3d157-3104-4225-8bc4-1b67f723acf4","order_by":4,"name":"Jing Jin","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Jing","middleName":"","lastName":"Jin","suffix":""},{"id":441055973,"identity":"6968dc9b-9ddb-4b0a-8579-b592c31777f6","order_by":5,"name":"Jing Yuan","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Jing","middleName":"","lastName":"Yuan","suffix":""},{"id":441055974,"identity":"0858b49a-fdfd-411c-9329-f2a2da52357c","order_by":6,"name":"Chizhou Liang","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Chizhou","middleName":"","lastName":"Liang","suffix":""},{"id":441055975,"identity":"649467ae-0b40-4d14-9b0b-607b8cdf54b1","order_by":7,"name":"Jianfei Lu","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Jianfei","middleName":"","lastName":"Lu","suffix":""},{"id":441055976,"identity":"c20f347c-aab2-4a06-a329-d0c1afaf20f0","order_by":8,"name":"Feng Cui","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Feng","middleName":"","lastName":"Cui","suffix":""},{"id":441055977,"identity":"515db542-971d-4319-8d74-3c71aadf9921","order_by":9,"name":"Jie Chen","email":"","orcid":"https://orcid.org/0009-0002-0956-1042","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Jie","middleName":"","lastName":"Chen","suffix":""}],"badges":[],"createdAt":"2025-01-03 08:35:52","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5756661/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5756661/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11104-025-07500-9","type":"published","date":"2025-05-14T15:58:01+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":80364707,"identity":"ecba8696-fc83-472e-adc5-19842e5312e8","added_by":"auto","created_at":"2025-04-11 05:01:05","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":32693,"visible":true,"origin":"","legend":"\u003cp\u003eThe impact of different treatments on hormone levels and defensive enzyme activities in pecan leaf tissues. (a): IAA content, (b): GA content, (c): SOD activity, (d): POD activity. Note: Different letters in the columns indicate significant differences (\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05), according to Duncan’s new multiple range test.\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5756661/v1/7513be8f1dfe8c37961e0778.jpg"},{"id":80363946,"identity":"a07954e9-af30-409b-9ae0-8ed2af75f345","added_by":"auto","created_at":"2025-04-11 04:53:04","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":43615,"visible":true,"origin":"","legend":"\u003cp\u003eThe effect of different treatments on the enzymatic activity in the rhizosphere soil of pecan. (a): Catalase activity, (b): Urease activity, (c): β-glucosidase activity. Note: Different letters in the columns indicate significant differences (\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05), according to Duncan’s new multiple range test.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5756661/v1/b59bc608ddc797bd2f0da820.jpg"},{"id":80364716,"identity":"7a2d6960-5afc-4d51-abf3-67c40b5479dd","added_by":"auto","created_at":"2025-04-11 05:01:05","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":38740,"visible":true,"origin":"","legend":"\u003cp\u003eThe impact of different treatments on the microbial abundance in the rhizosphere soil of pecan. (a): Number of fungi, (b): Number of bacteria, (c): Number of actinomycetes. Note: Different letters in the columns for each plant growth stage indicate significant differences (\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05), according to Duncan’s new multiple range test.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5756661/v1/ce803ee1666849c0f27f129d.jpg"},{"id":80364709,"identity":"93e684a8-be0e-4b1a-9814-a5bb8c88b0c9","added_by":"auto","created_at":"2025-04-11 05:01:05","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":36633,"visible":true,"origin":"","legend":"\u003cp\u003ePrincipal coordinate analysis (PCoA) of soil bacterial (a) and fungal (b) communities in the rhizosphere soil samples of pecan from different treatment groups.\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5756661/v1/d8e2dcce403e0678239d1309.jpg"},{"id":80363947,"identity":"a15618d2-71a0-4e21-9ce8-75229783a46b","added_by":"auto","created_at":"2025-04-11 04:53:04","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":64206,"visible":true,"origin":"","legend":"\u003cp\u003eDistribution of bacterial community abundance at the phylum level (a) and genus level (b) in rhizosphere soil of different treatment groups.\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5756661/v1/33a78834417dbf2e47105a0d.jpg"},{"id":80363949,"identity":"9720a1e4-2ab5-4068-843a-71120a3df87b","added_by":"auto","created_at":"2025-04-11 04:53:05","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":66676,"visible":true,"origin":"","legend":"\u003cp\u003eCluster heatmap analysis of bacterial community abundance at the genus level in the rhizosphere soil of different treatment groups.\u003c/p\u003e","description":"","filename":"6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5756661/v1/8c1ba7ca8db0ffc2a55f1685.jpg"},{"id":80363951,"identity":"1d46d25d-10c2-44ec-b152-8f7f8d8cbaf9","added_by":"auto","created_at":"2025-04-11 04:53:05","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":51959,"visible":true,"origin":"","legend":"\u003cp\u003eDistribution of fungal community abundance at the phylum level (A) and genus level (B) in the rhizosphere soil of different treatment groups.\u003c/p\u003e","description":"","filename":"7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5756661/v1/057c4ef11005fad49ed66e14.jpg"},{"id":80363954,"identity":"1018df04-9ba7-41de-95da-d0203b30b81a","added_by":"auto","created_at":"2025-04-11 04:53:05","extension":"jpg","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":50939,"visible":true,"origin":"","legend":"\u003cp\u003eCluster heatmap analysis of fungal community abundance at the genus level in the rhizosphere soil of different treatment groups.\u003c/p\u003e","description":"","filename":"8.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5756661/v1/0bafcda62acbaf9309e06a0c.jpg"},{"id":83067867,"identity":"f479d6cb-f22b-4d30-9757-48123065c9a6","added_by":"auto","created_at":"2025-05-19 16:07:16","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1454525,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5756661/v1/42de7313-a671-43e3-b507-138b17ed4bcc.pdf"}],"financialInterests":"","formattedTitle":"Exploring the potential of Trichoderma asperellum TCS007 on growth promotion of pecan seedlings as well as rhizosphere soil nutrients and microbial community","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eTrichoderma\u003c/em\u003e spp., ubiquitously present in nature, are a group of fungi that significantly promote plant growth (Chac\u0026oacute;n et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Tyśkiewicz et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Chen et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). These fungi colonize the rhizosphere of plants, enhancing their tolerance to biotic and abiotic stresses (Vargas et al. \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). They produce a variety of bioactive substances, including antibiotics and enzymes, which inhibit plant pathogens. By competing for nutrients and occupying space, they effectively suppress the growth of soil-borne pathogens, thereby protecting plants from disease (Garnica-Vergara et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Shi et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). \u003cem\u003eTrichoderma harzianum\u003c/em\u003e has been shown to grow 2.0 to 4.2 times faster than \u003cem\u003eBotrytis cinerea\u003c/em\u003e, thereby effectively inhibiting the latter's growth. This competitive exclusion not only deprives pathogens of essential resources but also physically blocks their access to plant tissues (Yao et al. \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Moreover, \u003cem\u003eTrichoderma\u003c/em\u003e induces systemic resistance in plants, strengthening their resilience to adverse conditions (Kashyap et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Gupta and Maya \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These functions make \u003cem\u003eTrichoderma\u003c/em\u003e an important biocontrol agent, widely used in agriculture.\u003c/p\u003e \u003cp\u003e \u003cem\u003eTrichoderma\u003c/em\u003e spp., have a significant positive impact on soil health. They promote the decomposition of organic matter in the soil, enhancing soil fertility and structure (Khan et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Halifu et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The metabolic activities of \u003cem\u003eTrichoderma\u003c/em\u003e increase the availability of nutrients such as nitrogen and phosphorus to plants (Abdenaceur et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Furthermore, \u003cem\u003eTrichoderma\u003c/em\u003e forms symbiotic relationships with plant roots, improving the efficiency of water and nutrient absorption by plants (Mehetre et al. 2015). They also improve the diversity of soil microbial communities, enhancing the ecological purification capabilities of soil (Mao and Jiang \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). These functions of \u003cem\u003eTrichoderma\u003c/em\u003e help maintain the balance of the soil ecosystem and improve its sustainable production capacity (Woo et al. \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Pinto et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Studies had shown that the application of \u003cem\u003eTrichoderma\u003c/em\u003e biofertilizers to the rhizosphere of corn could effectively increase the content of organic matter, nitrogen, phosphorus, and potassium in the soil. After spraying the suspension of \u003cem\u003eTrichoderma atroviride\u003c/em\u003e on corn at 15 and 25 days after germination, the soil ammonium nitrogen content increased by 15% (Fu et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Liu et al. (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) found that after applying a microbial fertilizer containing \u003cem\u003eTrichoderma guizhouense\u003c/em\u003e during the growth process of chili peppers, the soil urease activity significantly increased, and the transformation of soil organic nitrogen accelerated, increasing the available nitrogen content in the rhizosphere soil of chili peppers, promoting their nutrient absorption, and thus increasing the yield of chili peppers.\u003c/p\u003e \u003cp\u003ePecan was primarily found in the Tianmu Mountain area, located at the border of Zhejiang and Anhui provinces. Thriving in the understory of mountain slopes or valleys that were rich in humus, it was an economically important forest tree in China (Guo et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). In recent years, the excessive use of chemical fertilizers and pesticides, coupled with the long-term overexploitation of pecan forests, led to soil fertility degradation, frequent soil-borne diseases, and reduced yields, causing economic losses for farmers (Fang et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). There was an urgent need for a biofertilizer product that could improve the fertility of pecan forest land and was also friendly to the micro-ecological environment. \u003cem\u003eTrichoderma\u003c/em\u003e spp., as a green biological pesticide, had potential in environmental remediation and plant health, but their application in the pecan industry had been relatively understudied.\u003c/p\u003e \u003cp\u003eThis study explored the effects of the \u003cem\u003eTrichoderma asperellum\u003c/em\u003e TCS007 (isolated from Antarctic marine sediments) on the physiological and biochemical indicators of pecan plants, the physicochemical properties of rhizosphere soil, and soil enzymatic activity. High-throughput sequencing was used to analyze the diversity and structural composition of the microbial community in the rhizosphere soil of pecan, providing a theoretical basis for the development of TCS007 as a biofertilizer product, thereby contributing to the stable development of the pecan industry.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eExperimental materials and equipment\u003c/p\u003e \u003cp\u003eTested microbial strains and plants\u003c/p\u003e \u003cp\u003eThe \u003cem\u003eT. asperellum\u003c/em\u003e TCS007 was isolated from Antarctic marine sediments by our laboratory. The strain was preserved at the China General Microbiological Culture Collection Center (CGMCC), with the accession number CGMCC No.15677.\u003c/p\u003e \u003cp\u003eOne-year-old pecan seedlings, provided by the Panmugang Modern Forestry Demonstration Base, located in Lin'an District, Hangzhou City, Zhejiang Province, were transplanted into plastic pots filled with a mixture of peat soil/vermiculite/sand (2:1:1, v/v/v) as the growth substrate. After transplanting, the seedlings were placed in a greenhouse under 12 h photoperiod at 26\u0026deg;C and allowed to acclimatize for 30 days before use.\u003c/p\u003e \u003cp\u003eReagents and Culture Media\u003c/p\u003e \u003cp\u003ePlant hormone indole-3-acetic acid (IAA) and gibberellin (GA) assay kits were purchased from Nanjing Camilo Bioengineering Co., Ltd.; enzyme activity assay kits for soil catalase, urease, and β-glucosidase were purchased from Beijing Boxbio Science \u0026amp; Technology Co., Ltd., and soil genomic DNA extraction kits were purchased from Tiangen Biotech Co., Ltd.\u003c/p\u003e \u003cp\u003ePDA and PDB media (Li and He \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) were used for cultivating the TCS007. Tryptone Beef Broth, Martin medium, and Modified Gause's No. 1 medium (Li et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e1996\u003c/span\u003e) were used for the cultivation of bacteria, fungi, and actinomycetes from the soil samples.\u003c/p\u003e \u003cp\u003eExperimental Methods\u003c/p\u003e \u003cp\u003ePreparation of TCS007 spore suspension and liquid fermentation crude extract solution\u003c/p\u003e \u003cp\u003eFor spore suspension preparation; two agar plugs were punched from the edge of a revived TCS007 colony on PDA using a 9 mm diameter borer and inoculated into 100 mL PDB medium in a 250 mL flask for cultivation at 28\u0026deg;C at 180 rpm for 7 days. The liquid culture was then filtered through sterile gauze three times to remove the mycelia and debrises, obtained the fermentation filtrate of TCS007. The concentration of the spore suspension was determined using a hemocytometer and adjusted to 1.0 \u0026times; 10\u003csup\u003e6\u003c/sup\u003e and 1.0 \u0026times; 10\u003csup\u003e8\u003c/sup\u003e spores/mL for subsequent use.\u003c/p\u003e \u003cp\u003ePreparation of TCS007 liquid fermentation crude extract solution: The TCS007 fermentation filtrate was acquired as aforementioned. According to the method described by Liu et al. (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), the fermentation filtrate of TCS007 was extracted with an equal volume of ethyl acetate three times. The upper organic phase was collected and dried using a saturated sodium chloride solution and anhydrous sodium sulfate. After that, the extract was concentrated under reduced pressure using a rotary evaporator, redissolved in acetone, and collected in a centrifuge tube. Once the solvent had evaporated, the TCS007 organic phase extract was obtained. It was then diluted with sterile water to a concentration of 50 mg/mL for subsequent use.\u003c/p\u003e \u003cp\u003eExperimental Design and Seedlings Inoculation with \u003cem\u003eT. asperellum\u003c/em\u003e TCS007\u003c/p\u003e \u003cp\u003eThe pot experiment was conducted using a single-factor completely randomized block design with a total of five treatments (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Each treatment consisted of 20 seedlings as replicates, and the treatments were initiated subsequent to the seedlings that had been acclimatized over a period of 30 days. Each treatment was applied three times, with a 30-day interval between successive applications.\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\u003eTreatments used in this study\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\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 \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTest agent\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTreatment concentration\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTreatment method\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003esterile water\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eRoot irrigation was performed with a volume of 50 mL\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003esterilized PDB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTCS007 spore suspension\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.0 \u0026times; 10\u003csup\u003e8\u003c/sup\u003e spores/mL\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTCS007 spore suspension\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.0 \u0026times; 10\u003csup\u003e6\u003c/sup\u003e spores/mL\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTCS007 liquid fermentation crude extract\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e50 mg/mL\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\u003eDetermination of Physiological and Biochemical Indices of Seedlings\u003c/p\u003e \u003cp\u003eThree months after treatment, ten seedlings were randomly selected from each treatment for further analysis. Plant height and ground diameter were measured using a millimeter scale ruler. Chlorophyll content was determined by UV-2801 spectrophotometry (Hitachi, Ltd., Japan.) (Pan et al. 2017). Leaf IAA and GA content were quantified using enzyme-linked immunosorbent assay kits, employing a double-antibody sandwich method (Chen et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The activities of Superoxide dismutase (SOD) and Peroxidase (POD) enzymes were determined using the nitroblue tetrazolium method and the guaiacol method, respectively (Shahzad et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). With each treatment group replicated ten times.\u003c/p\u003e \u003cp\u003eDetermination of soil physicochemical properties, enzyme activities, and microbial counts\u003c/p\u003e \u003cp\u003eDuring the leaf expansion stage (mid-April), the new shoot growth stage (mid-July), and the defoliation stage (early October) of the pecan seedlings, rhizosphere soil samples from each treatment group were collected. The 20 pecan seedlings in a treatment were randomly divided into 5 groups. For each seedling, 12.5 g of soil sample was taken from the region rich in fine roots approximately 1 mm in diameter in the soil layer at a depth of 10\u0026ndash;30 cm (Zhang et al. \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The soil samples taken from the four seedlings assigned in the same group in one treatment were mixed thoroughly to form a single soil sample. In total, five soil samples as five replicates were collected for each treatment for subsequent processing. The soil samples were air-dried at 25\u0026deg;C, passed through a 1 mm sieve, collected in sterile sample bags, and then stored at 4\u0026deg;C for future use.\u003c/p\u003e \u003cp\u003eThe content of organic matter was determined using the dichromate volumetric method (Zhang and Fen 2023). The ammonium nitrogen content was determined using the titration method (Li et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2021b\u003c/span\u003e). The readily available phosphorus content was determined using the extraction-antimony molybdate colorimetric method (Li et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2021a\u003c/span\u003e). The content of ammonium nitrogen was determined using the titration method (Cheng, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The available potassium content was determined using the atomic absorption spectrophotometer with a flame photometer method (Guo et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The soil pH value was determined using the electrode method (Fu et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). With each treatment group replicated five times.\u003c/p\u003e \u003cp\u003eEnzyme activities of soil catalase, urease, and β-glucosidase were measured using assay kits from Beijing Boxbio Science \u0026amp; Technology Co., Ltd., following the manufacturer\u0026rsquo;s instructions. Catalase activity was assessed by monitoring the decomposition of H₂O₂, which reduced absorbance at 240 nm. One unit of catalase activity was defined as the amount of enzyme degrading 1 mmol H₂O₂ per gram of air-dried soil. Urease activity was determined by the formation of indophenol blue (absorbance at 630 nm) from NH₃-N generated by urea hydrolysis. One unit of urease activity corresponded to the production of 1 \u0026micro;g NH₃-N per gram of soil. β-Glucosidase activity was measured by the hydrolysis of p-nitrophenyl-β-D-glucopyranoside to p-nitrophenol, with absorbance at 400 nm. One unit of β-glucosidase activity was defined as the amount of enzyme generating 1 \u0026micro;mol p-nitrophenol per gram of soil. With each treatment group replicated five times.\u003c/p\u003e \u003cp\u003eThe numbers of bacteria, fungi, and actinomycetes in the soil samples from each treatment group were determined using the dilution plating method. Briefly, in a 250 mL Erlenmeyer flask, 50 mL of sterile water was added. Then, 5 g of fresh soil sample was introduced into the flask and agitated at room temperature for 10 min. Subsequently, 1 mL of the soil suspension was transferred to 9 mL of sterile water, and the resulting suspension was serially diluted by a factor of 10 up to 10\u003csup\u003e\u0026minus;\u0026thinsp;6\u003c/sup\u003e. Based on preliminary experiments, for this trial, fungi were diluted between 10\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e and 10\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, while bacteria and actinomycetes were diluted between 10\u003csup\u003e\u0026minus;\u0026thinsp;5\u003c/sup\u003e and 10\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e. An aliquot of 50 \u0026micro;L of the diluted solution was spread evenly onto the culture medium with each dilution repeated three times (Li and He \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Tryptone Soya Broth agar, Martin medium, and Modified Gause's No. 1 medium (Li et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e1996\u003c/span\u003e) were utilized for the isolation and enumeration of bacteria, fungi, and actinomycetes, respectively. With each treatment group replicated five times.\u003c/p\u003e \u003cp\u003eConstruction of Internal Transcribed Spacer (ITS) and 16S rRNA Gene Libraries and Subsequent High-Throughput Sequencing Analysis\u003c/p\u003e \u003cp\u003eDuring the leaf expansion stage of the pecan seedlings, soil microbial DNA was extracted from the five soil samples in each treatment group. The quality of the extracted DNA was assessed using 1.5% agarose gel electrophoresis and a spectrophotometer. The V3-V4 region of the bacterial 16S rRNA gene and the ITS1 region of the fungal ITS sequence were amplified using the universal primers 338F (5'-ACTCCTACGGGAGGCAGCAG-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3'), as well as ITS1F (5'-CTTGGTCATTTAGAGGAAGTAA-3') and ITS2R (5'-GCTGCGTTCTTCATCGATGC-3'). PCR amplification was conducted in a 20 \u0026micro;L reaction system using TransStart FastPfu DNA Polymerase and TaKaRa rTaq DNA Polymerase. After amplification, the PCR products were recovered through purification, detection, and quantification steps. Subsequently, the amplified products were mixed in equimolar concentrations to form a single sequencing library. The constructed library was quality checked, and qualified libraries were sequenced using the Illumina MiSeq sequencing platform (PE300).\u003c/p\u003e \u003cp\u003eFLASH v1.2.7 software was used to assemble reads from each sample based on overlaps, yielding raw Tag data (Magoč et al. 2011). Trimmomatic v0.33 software was employed to filter the raw Tag data, resulting in high-quality Tag data (Bolger et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). UCHIME v4.2 software was utilized to identify and remove chimeric sequences, obtaining the final data.\u003c/p\u003e \u003cp\u003eFor soil microbial genomic data, Operational Taxonomic Units (OTUs) clustering analysis was conducted using USEARCH software. Biostatistical analysis on OTUs at 97% similarity level was performed (Edgar et al. 2013). The clustered OTUs were then used for various analyses: alpha diversity analysis (ACE index, Chao1 index, Shannon index, Simpson index, and Coverage index), beta diversity analysis, and species composition and difference analysis. Alpha diversity indices were calculated under different random samplings using Mothur software, while beta diversity analysis was conducted with QIIME software. Non-metric multidimensional scaling (NMDS) analysis and heatmap analysis were executed using R software, and the ANOSIM test was applied for statistical comparisons. All tables were prepared with Excel 2021 software, and the analyses were performed on the Majorbio Cloud Platform (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ewww.majorbio.com\u003c/span\u003e\u003cspan address=\"http://www.majorbio.com\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) in Shanghai.\u003c/p\u003e \u003cp\u003eStatistical Analysis and Data Interpretation\u003c/p\u003e \u003cp\u003ePhysiological and biochemical data of plants and soil were statistically analyzed using Excel 2021 software. To determine differences in physiological and biochemical indices of pecan seedlings, soil physicochemical properties, and soil enzymatic activities, one-way analysis of variance (ANOVA) and Duncan's test within IBM SPSS 22.0 software (IBM Corporation, New York, NY, USA) were employed.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe Impact of TCS007 on the Physiological and Biochemical Indicators of Pecan Plant\u003c/p\u003e \u003cp\u003ePlant height, chlorophyll content, and ground diameter were measured for each treatment group. Compared to the control group A, treatment group C showed a height increase of 31.8%, a chlorophyll content increase of 225.8%, and a ground diameter increase of 329.6%. Treatment group E exhibited increases of 19.3%, 56.3%, and 29.2%, respectively, in height, chlorophyll content, and groun d diameter, while treatment group B showed no significant differences in all three indicators. The experimental results indicated that both the TCS007 spore suspension and crude extract promoted the growth of pecan plants, with the optimal growth-promoting effect observed at a concentration of 1.0 \u0026times; 10\u003csup\u003e8\u003c/sup\u003e spores/mL for the TCS007 spore suspension (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe impact of \u003cem\u003eTrichoderma\u003c/em\u003e inoculation on seedling biomass\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\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 \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment groups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePlant Height (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGround diameter (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChlorophyll content (mg\u0026middot;g\u003csup\u003e-1\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19 d\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.82 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07 d\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.84 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.96 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17.38\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.64 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eNote: Different letters in the columns indicate significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), according to Duncan\u0026rsquo;s new multiple range test.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eIn terms of phytohormone content, the C treatment group showed an increase of 60.0% in IAA and 60.8% in GA in the leaf tissues, compared to the A treatment group. The E treatment group exhibited increases of 26.2% in IAA and 31.4% in GA, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea-b). Regarding defense enzymes, the C treatment group had a 53.5% increase in superoxide dismutase (SOD) and a 140.0% increase in peroxidase (POD) enzyme activities in the leaf tissues. The treatment of group D showed even higher increases by 69.5% and 166.1% for SOD and POD, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec-d). The B treatment group did not exhibit significant differences in either phytohormone content or defense enzyme activities. The experimental results suggested that both the TCS007 spore suspension and crude extract could enhance hormone content and defense enzyme activities in the leaf tissues of pecan plants, with the most pronounced effects observed at a TCS007 spore suspension concentration of 1.0 \u0026times; 10\u003csup\u003e8\u003c/sup\u003e spores/mL.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe Impact of TCS007 on the Physicochemical Properties of Soil in the Rhizosphere of pecan\u003c/p\u003e \u003cp\u003eDuring the leaf expansion, shoot growth, and defoliation stages of pecan seedlings, the organic matter content in the treatment of group C increased by 19.4%, 17.0%, and 10.9%, respectively, the ammonium nitrogen content increased by 20.0%, 21.3%, and 10.3%, respectively, and the available phosphorus content increased by 23.7%, 16.7%, and 17.7%, respectively. The soil pH values were elevated to 7.53, 6.96, and 6.76, respectively. In the treatment of group D, the organic matter content increased by 12.7%, 13.2%, and 6.3%, the ammonium nitrogen content increased by 17.3%, 15.3%, and 8.0%, and the available phosphorus content increased by 13.7%, 14.4%, and 15.2%, with soil pH values elevated to 7.32, 6.84, and 6.62, respectively. However, there were no significant differences in available potassium (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The experimental results indicated that the TCS007 spore suspension significantly increased the organic matter content, ammonium nitrogen content, available phosphorus content, and soil pH in the rhizosphere soil of pecan, while the crude extract treatment group showed no significant differences.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe influence of different treatments on the physicochemical properties of the rhizosphere soil during various growth stages of pecan\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\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 \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePecan growth stages\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreatment groups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOrganic matter content/%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAmmonium Nitrogen /mg\u0026middot;kg\u003csup\u003e-1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAvailable Phosphorus /mg\u0026middot;k\u003csup\u003e-1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAvailable Potassium /mg\u0026middot;kg\u003csup\u003e-1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003epH value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eLeaf expansion stage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.73 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e44.57\u0026thinsp;\u0026plusmn;\u0026thinsp;2.38 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.92\u0026thinsp;\u0026plusmn;\u0026thinsp;2.19 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e54.63\u0026thinsp;\u0026plusmn;\u0026thinsp;3.25 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.32\u0026thinsp;\u0026plusmn;\u0026thinsp;5.36 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.63\u0026thinsp;\u0026plusmn;\u0026thinsp;1.83 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e46.76\u0026thinsp;\u0026plusmn;\u0026thinsp;2.03 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e23.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.97 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e53.96\u0026thinsp;\u0026plusmn;\u0026thinsp;1.64 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.82 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e53.49\u0026thinsp;\u0026plusmn;\u0026thinsp;1.97 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e28.36\u0026thinsp;\u0026plusmn;\u0026thinsp;1.87 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e54.38\u0026thinsp;\u0026plusmn;\u0026thinsp;2.33 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7.53\u0026thinsp;\u0026plusmn;\u0026thinsp;3.28 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.62 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e52.28\u0026thinsp;\u0026plusmn;\u0026thinsp;1.23 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e26.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e54.20\u0026thinsp;\u0026plusmn;\u0026thinsp;1.47 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7.32\u0026thinsp;\u0026plusmn;\u0026thinsp;2.08 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.52\u0026thinsp;\u0026plusmn;\u0026thinsp;1.32 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e43.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.28 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.65 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e54.32\u0026thinsp;\u0026plusmn;\u0026thinsp;1.48 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.49\u0026thinsp;\u0026plusmn;\u0026thinsp;1.38 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eShoot growth stage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.42\u0026thinsp;\u0026plusmn;\u0026thinsp;1.48 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e45.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e23.64\u0026thinsp;\u0026plusmn;\u0026thinsp;1.93 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e55.49\u0026thinsp;\u0026plusmn;\u0026thinsp;2.39 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.44\u0026thinsp;\u0026plusmn;\u0026thinsp;2.01 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e46.18\u0026thinsp;\u0026plusmn;\u0026thinsp;1.63 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.78 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e55.29\u0026thinsp;\u0026plusmn;\u0026thinsp;1.47 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.37\u0026thinsp;\u0026plusmn;\u0026thinsp;1.84 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.83\u0026thinsp;\u0026plusmn;\u0026thinsp;1.04 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e54.88\u0026thinsp;\u0026plusmn;\u0026thinsp;2.31 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e27.58\u0026thinsp;\u0026plusmn;\u0026thinsp;1.27 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e54.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.49 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.74\u0026thinsp;\u0026plusmn;\u0026thinsp;1.23 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e52.16\u0026thinsp;\u0026plusmn;\u0026thinsp;1.71 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e27.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.85 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e53.26\u0026thinsp;\u0026plusmn;\u0026thinsp;1.39 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.84\u0026thinsp;\u0026plusmn;\u0026thinsp;1.63 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.73 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e45.87\u0026thinsp;\u0026plusmn;\u0026thinsp;1.02 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e23.17\u0026thinsp;\u0026plusmn;\u0026thinsp;1.38 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e52.93\u0026thinsp;\u0026plusmn;\u0026thinsp;3.19 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.76 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eDefoliation stage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.39\u0026thinsp;\u0026plusmn;\u0026thinsp;1.76 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e42.33\u0026thinsp;\u0026plusmn;\u0026thinsp;1.74 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.95\u0026thinsp;\u0026plusmn;\u0026thinsp;1.47 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e50.88\u0026thinsp;\u0026plusmn;\u0026thinsp;2.38 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.27\u0026thinsp;\u0026plusmn;\u0026thinsp;1.68 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.41\u0026thinsp;\u0026plusmn;\u0026thinsp;1.27 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e42.89\u0026thinsp;\u0026plusmn;\u0026thinsp;2.18 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.16\u0026thinsp;\u0026plusmn;\u0026thinsp;1.29 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e48.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.33\u0026thinsp;\u0026plusmn;\u0026thinsp;3.94 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e46.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.87 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e49.73\u0026thinsp;\u0026plusmn;\u0026thinsp;1.20 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.76\u0026thinsp;\u0026plusmn;\u0026thinsp;2.06 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.54\u0026thinsp;\u0026plusmn;\u0026thinsp;1.32 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e45.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.97 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25.29\u0026thinsp;\u0026plusmn;\u0026thinsp;1.94 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e49.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.68 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.62\u0026thinsp;\u0026plusmn;\u0026thinsp;1.83 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.31\u0026thinsp;\u0026plusmn;\u0026thinsp;1.76 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e41.97\u0026thinsp;\u0026plusmn;\u0026thinsp;1.22 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.99 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e51.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.19\u0026thinsp;\u0026plusmn;\u0026thinsp;6.93 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003eNote: Different letters in the columns indicate significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), according to Duncan\u0026rsquo;s new multiple range test.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe Impact of TCS007 on the Enzymatic Activity of Rhizosphere Soil in Pecan\u003c/p\u003e \u003cp\u003eDuring the leaf expansion stage of the pecan, the activities of soil catalase, urease, and β-glucosidase in the C treatment group significantly increased by 55.2%, 78.4%, and 62.0%, respectively, compared to the A treatment group. The E treatment group also showed increases in soil enzyme activities of 32.8%, 22.4%, and 96.3%. During the shoot growth stage, the C treatment group showed an increase in soil enzyme activities of 46.1%, 37.6%, and 44.1%, while the E treatment group's increases were 36.5%, 9.0%, and 65.8%. By the time of defoliation stage, the C treatment group's soil enzyme activities had further increased by 30.5%, 20.8%, and 23.2%, the E treatment group's activities increased the soil enzymatic activity by 25.5%, 15.9%, and 22.7%, and the B treatment group showed no significant difference. The experimental results indicated that the spore suspension and crude extract of TCS007 could effectively enhance soil enzyme activity, with the crude extract showing the most pronounced effect (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe Influence of TCS007 on the Culturable Microbial Population in the Rhizosphere Soil of Pecan\u003c/p\u003e \u003cp\u003eDuring the leaf expansion stage, compared to the A treatment group, the microbial population in the rhizosphere soil of the C treatment group significantly increased, while in the E treatment group, the number of fungi decreased by 69.2%, bacteria by 57.8%, and actinomycetes by 66.7%. During the shoot growth stage, compared to the A treatment group, the microbial population in the rhizosphere soil of the C treatment group significantly increased again, with a 49.6% reduction in fungal numbers, a 21.1% increase in bacterial numbers, and a significant 79.3% decrease in actinomycetes in the E treatment group. In the defoliation stage, compared to the A treatment group, the number of fungi and bacteria in the rhizosphere soil of the C treatment group significantly increased, while the actinomycetes decreased by 7.5%. In the E treatment group, the numbers of fungi and actinomycetes decreased by 39.2% and 16.4%. Respectively. In the B treatment group, the numbers of fungi and bacteria only significantly increased during the leaf expansion stage, with no significant changes in the microbial population in the rhizosphere soil during other stages. The experimental results indicated that during the growth process of pecan plants, the number of fungi in the rhizosphere soil first decreased and then increased after treatment with TCS007 spore suspension (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea), bacteria decreased and then stabilized (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb), and actinomycetes significantly decreased (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec). After treatment with TCS007 crude extract, the number of fungi in the rhizosphere soil significantly decreased (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea), the number of bacteria significantly increased after the leaf expansion stage (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb), and the number of actinomycetes did not significantly change (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec). The spore suspension and crude extract of TCS007 both significantly affected the number of soil microorganisms, with the spore suspension of TCS007 having the most pronounced effect (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe Influence of TCS007 on the Microbial Composition of the Rhizosphere Soil in Pecan\u003c/p\u003e \u003cp\u003eMicrobial Sequencing Data Analysis\u003c/p\u003e \u003cp\u003eThe statistics of the sequencing data after quality control, filtering, and assembly for the five treatment groups are presented in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. The effective bacterial sequences were 28089, 21237, 26931, 25779, and 24914; and the effective fungal sequences were 49472, 49101, 45903, 39691, and 42696. The sequencing quality, measured by the Q30 score, ranged from 97\u0026ndash;99%, while the Q20 score ranged from 99\u0026ndash;99.5% (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSummary of basic sequencing data for each sample\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\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 \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMicroorganisms\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eShortest tags\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLongest tags\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMean tags\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eEffective tags\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eTotal bases\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eBacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e504\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e419\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e28089\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e21438243\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e317\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e527\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e417\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e21237\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e21312593\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e317\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e497\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e418\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e26931\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e21395076\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e319\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e456\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e419\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e25779\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e19405297\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e270\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e469\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e418\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e24914\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e19781631\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eFungi\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e141\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e526\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e237\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e49472\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12587811\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e141\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e533\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e232\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e49101\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12441487\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e142\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e384\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e245\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e45903\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12569760\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e143\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e522\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e237\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e39691\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e10209814\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e142\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e514\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e236\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e42696\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e11074245\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\u003eMicrobial Community Diversity Analysis\u003c/p\u003e \u003cp\u003eAlpha diversity indices analysis revealed that the coverage indices for all treatment groups were close to 1, indicating that the sequencing results could accurately reflect the actual conditions of the soil samples tested. The ACE and Chao indices were indicative of the richness of soil microbial communities, while the Simpson and Shannon indices reflected the diversity of these communities. Among bacterial samples, the analysis of Chao1 and ACE indices across treatment groups revealed that the bacterial community richness in treatment groups C and D was significantly higher than in groups A and B (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), with the bacterial community richness in pecan rhizosphere soil increasing as the concentration of TCS007 spores increased. Analysis of the Simpson and Shannon indices showed that the bacterial community diversity in treatment group C was significantly higher than in other groups (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), with no significant difference in diversity between groups B and D compared to group A. After treatment with group E, the bacterial community richness and diversity in the pecan rhizosphere were significantly reduced compared to other treatment groups (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). In the case of fungal samples, the analysis of Chao1 and ACE indices demonstrated that the richness of fungal communities in treatment groups C and D was significantly higher than in other groups (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Analysis of the Shannon and Simpson indices indicated that the fungal community diversity in groups C and D was significantly higher than in group A (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). However, the richness and diversity of fungal communities in group E were significantly lower than in other treatment groups (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe impact of various treatments on the diversity indices of the microbial community\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\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 \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMicroorganisms\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eShannon Index\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSimpson Index\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eChao Index\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eACE Index\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCoverage %\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eBacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1579.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1591.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1634.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1666.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1708.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1725.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1699.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1714.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1315.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1337.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eFungi\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e515.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e515.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e551.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e553.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e630.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e632.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e568.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e569.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e414.17e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e416.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003eNote: Different letters in the columns indicate significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), according to Duncan\u0026rsquo;s new multiple range test.\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003eMicrobial Community Compositional Differences Analysis\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eBeta diversity analysis revealed that compared to the treatment group A, the soil bacterial community compositions of the other treatment groups showed significant differences, with the most pronounced difference observed between treatment group C and group A, while treatment groups D and E exhibited a higher similarity in their bacterial community compositions (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ea). In comparison to treatment group A, the soil fungal community compositions of treatment groups C and D were significantly different, whereas the fungal community composition of treatment group E showed a higher similarity to that of treatment group A (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe analysis of soil microbial community compositional differences revealed that the structure of the community in different samples was known at various taxonomic levels, such as domain, kingdom, phylum, class, order, family, genus, and species. The bacterial community in the rhizosphere soil of pecan was primarily composed of the phyla \u003cem\u003eActinobacteria\u003c/em\u003e, \u003cem\u003eProteobacteria\u003c/em\u003e, \u003cem\u003eAcidobacteria\u003c/em\u003e, and \u003cem\u003eChloroflexi\u003c/em\u003e, with \u003cem\u003eActinobacteria\u003c/em\u003e, \u003cem\u003eProteobacteria\u003c/em\u003e, and \u003cem\u003eAcidobacteria\u003c/em\u003e being the dominant. Compared to treatment A, treatment C increased the abundance of \u003cem\u003eActinobacteria\u003c/em\u003e by 36.19% and significantly increased the abundance of \u003cem\u003eFirmicutes\u003c/em\u003e by 301.57%. In contrast, the abundance of \u003cem\u003eAcidobacteria\u003c/em\u003e and \u003cem\u003eChloroflexi\u003c/em\u003e in the rhizosphere soil bacterial community decreased by 45.49% and 34.92%, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ea). Additionally, treatment C reduced the abundance of the \u003cem\u003eAcidobacteriaceae\u003c/em\u003e and \u003cem\u003eRB41\u003c/em\u003e genera in the bacterial community by 46.25% and 61.03%, respectively, while the abundance of the genera \u003cem\u003eNocardioides\u003c/em\u003e, \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003eMicrococcaceae\u003c/em\u003e, and \u003cem\u003eAgromyces\u003c/em\u003e increased, with \u003cem\u003eBacillus\u003c/em\u003e showing a significant increase of 377.65%. However, treatments B, D, and E did not show significant differences in the abundance of the bacterial community in the rhizosphere soil compared to treatment A (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eb, Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe fungal community in the rhizosphere soil of pecan was predominantly composed of the phyla \u003cem\u003eAscomycota\u003c/em\u003e, \u003cem\u003eMortierellomycota\u003c/em\u003e, and \u003cem\u003eBasidiomycota\u003c/em\u003e, with \u003cem\u003eAscomycota\u003c/em\u003e and \u003cem\u003eMortierellomycota\u003c/em\u003e being the dominant. Compared to the A treatment group, the abundance of \u003cem\u003eAscomycota\u003c/em\u003e in the C treatment group increased by 23.72%, while the abundance of \u003cem\u003eMortierellomycota\u003c/em\u003e and \u003cem\u003eBasidiomycota\u003c/em\u003e decreased by 41.18% and 21.87%, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003ea). Furthermore, the abundance of the genera \u003cem\u003eTrichoderma\u003c/em\u003e and \u003cem\u003eApiotrichum\u003c/em\u003e in the C treatment group significantly increased, while other fungi were generally inhibited. Notably, the genera \u003cem\u003eMortierella\u003c/em\u003e, \u003cem\u003eNeocosmospora\u003c/em\u003e, \u003cem\u003eFusarium\u003c/em\u003e, \u003cem\u003eChaetomium\u003c/em\u003e, \u003cem\u003eTalaromyces\u003c/em\u003e, \u003cem\u003eAspergillus\u003c/em\u003e, \u003cem\u003eCladosporium\u003c/em\u003e, and \u003cem\u003eAcremonium\u003c/em\u003e showed the most pronounced inhibition. Specifically, the abundance of \u003cem\u003eMortierella\u003c/em\u003e, \u003cem\u003eNeocosmospora\u003c/em\u003e, \u003cem\u003eFusarium\u003c/em\u003e, and \u003cem\u003eCladosporium\u003c/em\u003e decreased by 41.10%, 45.77%, 55.71%, and 46.15%, respectively. However, the B, D, and E treatment groups showed no significant differences (Figs.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eb and \u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eAs a commonly used biocontrol agent, \u003cem\u003eTrichoderma\u003c/em\u003e species exhibit a range of efficacy. Upon application, \u003cem\u003eTrichoderma\u003c/em\u003e played a crucial role in the rhizosphere by competing against other microbes, particularly plant pathogens (Ferreira and Musumeci \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). This competition was multifaceted and included direct antagonism, nutrient and space competition, and the production of antifungal compounds (Manzar et al \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Additionally, \u003cem\u003eTrichoderma\u003c/em\u003e could outcompete pathogens for nutrients and space by rapidly colonizing plant roots and the surrounding soil (Tyśkiewicz et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). This competition not only suppressed the growth of pathogens but also promoted plant health by enhancing nutrient uptake and inducing systemic resistance in plants.(Dutta et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Then, it produces plant growth hormones that enhance the solubility of nutrients in the soil, improves rhizosphere microecology, and thereby facilitates nutrient uptake by plants, promoting growth and increasing yield (Contreras-Cornejo et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Wei et al. 2024). Additionally, \u003cem\u003eTrichoderma\u003c/em\u003e induces the production of defense enzymes in plants, such as peroxidase (POD) and superoxide dismutase (SOD), which play a crucial role in the plant resistance to stress and diseases (Ahmad et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Zhang et al. \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Pacheco-Trejo et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). \u003cem\u003eTrichoderma\u003c/em\u003e elicits plant immune responses through multiple mechanisms. It secretes microbe-associated molecular patterns (MAMPs) recognized by plant cell surface pattern recognition receptors (PRRs), triggering basal immunity (Alfiky and Weisskopf \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Our study revealed that the TCS007 spore suspension and crude extracts significantly promoted the growth of pecan seedlings and enhanced chlorophyll content in leaves. Concurrently, there was a notable increase in the levels of plant hormones (IAA, GA) and the activity of defense enzymes (POD, SOD) in the leaves, with the optimal effect observed at a spore suspension concentration of 1\u0026times;10\u003csup\u003e8\u003c/sup\u003e spores/mL. Studies showed that \u003cem\u003eTrichoderma\u003c/em\u003e strains isolated from various regions could secrete IAA, enhancing the photosynthetic rate and promoting the growth of plants such as cucumber and corn (Vinale et al. \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2008a\u003c/span\u003e; Vargas et al. \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Zhang et al. (\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) found that \u003cem\u003eTrichoderma harzianum\u003c/em\u003e significantly increased the plant height, basal diameter, and fresh weight of \u003cem\u003eMalus hupehensis\u003c/em\u003e Rehd. Moreover, \u003cem\u003eTrichoderma\u003c/em\u003e species were capable of producing a variety of secondary metabolites, including 6-pentyl-2H-pyran-2-one (6PP), Koninginin A5, gliotoxin, and viridin, which played significant signaling roles in plant-microbe interactions (Forde et al. 2014; Garnica-Vergara et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Moisan et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). It was demonstrated that \u003cem\u003eTrichoderma atroviride\u003c/em\u003e could produce at least 25 types of volatile organic compounds, encompassing alcohols, ketones, alkanes, ethylene, monoterpenes, and sesquiterpenes, among which gaseous ethylene acted as a core signaling molecule in \u003cem\u003eTrichoderma\u003c/em\u003e-plant interactions (Lee et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Estrada-Rivera et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Vinale et al. (\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2008b\u003c/span\u003e) observed growth promotion in etiolated pea stems treated with the main secondary metabolites produced by different \u003cem\u003eTrichoderma\u003c/em\u003e strains, harzianic acid, and 6PP.\u003c/p\u003e \u003cp\u003eSoil nutrients, often existing in sparingly soluble or insoluble forms, impeded nutrient cycling, highlighting the importance of enhancing nutrient availability. \u003cem\u003eTrichoderma\u003c/em\u003e species, through the secretion of organic acids, activated soil nutrients and promoted plant absorption, playing a crucial role in rehabilitating degraded soils (Sindhu et al. \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Singh et al. \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Our study demonstrated that TCS007 spore suspension significantly increased organic matter, ammonium nitrogen, available phosphorus content, and pH value in the rhizosphere soil of pecan, with the optimal effect at a concentration of 1\u0026times;10\u003csup\u003e8\u003c/sup\u003e spores/mL. Asghar and Kataoka. (2021) found that soil phosphatase activity had increased by more than 10% after inoculation with \u003cem\u003eTrichoderma\u003c/em\u003e RW309 for two months, accelerating the transformation of organic phosphorus compounds or inorganic phosphates in the soil. The physiological metabolism of different microbial communities was closely related to the production of soil enzymes, which could reflect the presence and activity of corresponding functional microbes (Mao and Jiang. 2021; Jia et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Aljeddani et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). \u003cem\u003eTrichoderma\u003c/em\u003e species rapid colonization capability enhanced rhizosphere-soil contact, promoting the secretion of extracellular enzymes such as catalase, urease, and β-glucosidase, as well as organic acids (Solomon et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Catalase served as an indicator of soil microecology, urease reflected the soil's nitrogen supply capacity, and β-glucosidase provided a carbon source for soil microbes (Woese et al. 1987; Stott et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Su et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Our results indicated that TCS007 spore suspension and crude extracts effectively enhanced soil enzyme activity, with the most significant effect observed at a concentration of 50 mg/L for the crude extracts. In a cabbage pot experiment, the treatment with \u003cem\u003eTrichoderma\u003c/em\u003e increased soil transformation enzyme activity by 45%, effectively promoting nutrient transformation and improving the soil ecological environment (Shi et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eForest soil microbes played a pivotal role in ecosystems, encompassing soil formation, development, and the maintenance of ecological balance. The ratio of soil bacteria, fungi, and actinomycetes served as a vital indicator of soil fertility, with their quantities showing a significant positive correlation with soil nutrients and crop yields (Hang et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Increasing the populations of rhizosphere bacteria and actinomycetes aided in enhancing soil nitrogen and phosphorus content, promoting plant root development and nutrient absorption (M\u0026auml;kip\u0026auml;\u0026auml; et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Xiong et al. \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Harsonowati et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Chen et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Soil microbial diversity and richness are considered critical for the integrity, function, and long-term sustainability of soil ecosystems. The latter are usually reduced by agricultural perturbations (Fu et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Our study found that TCS007 spore suspension significantly increased the number of culturable bacteria and actinomycetes in the rhizosphere soil of pecan during the leaf expansion and shoot growth stage, particularly the beneficial microbes \u003cem\u003eNocardia\u003c/em\u003e and \u003cem\u003eBacillus\u003c/em\u003e, enhanced the abundance of bacterial community richness and bacterial community diversity in the soil. Concurrently, TCS007 spore suspension increased the abundance of \u003cem\u003eTrichoderma\u003c/em\u003e and \u003cem\u003eApiotrichum\u003c/em\u003e fungi, suppressing the growth of plant pathogens such as \u003cem\u003eFusarium\u003c/em\u003e, \u003cem\u003eColletotrichum\u003c/em\u003e, and \u003cem\u003eMagnaporthe oryzae\u003c/em\u003e, thus promoting the growth of beneficial fungal communities and inhibiting pathogenic fungi. Consequently treating soil with TCS007 spore suspension would altering the uniformity and richness of soil species, and altering the soil\u0026rsquo;s microbial diversity. Fu et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) study indicated that \u003cem\u003eTrichoderma\u003c/em\u003e treatment impacts bacterial diversity in the rhizosphere soil of maize. The diversity of the bacterial community in soil following \u003cem\u003eTrichoderma\u003c/em\u003e treatment was lower than that in the control, untreated, soil. This might be associated with the fact that Trichoderma promotes plant growth and plant nutrient absorption from the soil. Hou et al. (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) demonstrated that \u003cem\u003eTrichoderma harzianum\u003c/em\u003e TH62 effectively inhibited soil-borne pathogens such as \u003cem\u003eFusarium oxysporum\u003c/em\u003e and \u003cem\u003eAlternaria alternata\u003c/em\u003e. In addition, \u003cem\u003eT. harzianum\u003c/em\u003e ESALQ-1306 and \u003cem\u003eT. asperellum\u003c/em\u003e BRM-29104 had a minor impact on soil fungal abundance, they shifted the soil microbial community from fungi to bacteria, maintaining microbial balance and reducing crop diseases and pests (Cordier and Alabouvette. 2009; Zhang et al. \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Silva et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). These results indicated that TCS007 spore suspension could regulate soil microbial communities, enhancing soil fertility and crop health. Future research should focus on elucidating the specific mechanisms underlying the beneficial effects of \u003cem\u003eT. asperellum\u003c/em\u003e TCS007 on pecan plants and their rhizosphere environment. Long-term field trials are needed to assess the sustained impact of TCS007 on soil health and plant growth under varying environmental conditions. Additionally, studies should explore the potential synergistic effects of TCS007 with other soil amendments or agricultural practices. Investigating the genetic and biochemical pathways involved in the interactions between TCS007 and pecan plants could provide valuable insights into optimizing its application as a biological fungal fertilizer.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eIn a pioneering effort, our research team has introduced the marine-derived strain \u003cem\u003eT. asperellum\u003c/em\u003e TCS007 to the economically important tree species pecan. This study demonstrated that treatment with TCS007 spore suspension or crude extract promoted the growth of pecan seedlings, with significant increases in plant height, ground diameter, and chlorophyll content compared to the CK. Additionally, the contents of IAA and GA in the leaves, as well as the activities of SOD and POD enzymes, were significantly enhanced. The best growth-promoting effect was observed when the TCS007 spore suspension was applied at a concentration of 1.0 \u0026times; 10\u003csup\u003e8\u003c/sup\u003e spores/mL.\u003c/p\u003e \u003cp\u003eIn addition, the levels of organic matter, ammonium nitrogen, and available phosphorus content in the rhizosphere soil, as well as the activities of soil catalase, urease, and β-glucosidase, were significantly higher in the treatment groups than those in the CK, and the soil pH shifted from slightly acidic to slightly alkaline after treatment. The soil microbial communities of pecan seedlings treated with TCS007 were analyzed during the leaf expansion stage, high-throughput sequencing revealed that the abundance of beneficial microbes, such as \u003cem\u003eNocardia\u003c/em\u003e, \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003eMicrococcus\u003c/em\u003e, and \u003cem\u003eMycobacterium\u003c/em\u003e in the bacterial community and \u003cem\u003eTrichoderma\u003c/em\u003e and \u003cem\u003ePenicillium\u003c/em\u003e in the fungal community, significantly increased, while the abundance of \u003cem\u003eAcidobacterium\u003c/em\u003e and \u003cem\u003eRB41\u003c/em\u003e in bacterial community and other fungal genera decreased in the group treated with TCS007 spore suspension.\u003c/p\u003e \u003cp\u003eIn summary, the TCS007 spore suspension could effectively promote the growth and development of pecan plants, improved the stability of the rhizosphere soil microecology of pecan, and increased the content of soil nutrients. It had a positive effect on the prevention and control of soil-borne diseases in pecan and the regulation of soil pH. These results indicated that \u003cem\u003eT. asperellum\u003c/em\u003e TCS007 had great potential in being developed into a biological fungal fertilizer product for pecan.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eDeclarations\u003c/h2\u003e \u003cp\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e\n\u003ch3\u003eFounding\u003c/h3\u003e\n\u003cp\u003eThis study was supported by the Science and Technology Collaboration Program of Zhejiang Province (2023SNJF035), the National Key Research and Development Program of China (2022YFD1700400).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbdenaceur, R., Farida, Bt., Mourad, D., Rima, H., Zahia, O., Fatma, S. 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Chinese Journal of Pesticide Science\u003cem\u003e.\u003c/em\u003e 2020, 22(05), 801-807. doi: 10.16801/j.issn.1008-7303.2020.0075.\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":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"plant-and-soil","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"plso","sideBox":"Learn more about [Plant and Soil](https://www.springer.com/journal/11104)","snPcode":"11104","submissionUrl":"https://submission.nature.com/new-submission/11104/3","title":"Plant and Soil","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Biofertilizers, Forest trees, Soil physicochemical properties, soil enzyme activities, microbial diversity","lastPublishedDoi":"10.21203/rs.3.rs-5756661/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5756661/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eAims\u003c/h2\u003e \u003cp\u003ePecan (\u003cem\u003eCarya cathayensis\u003c/em\u003e Sarg.) is an important forest trees in China, the application of chemical pesticides for disease control has caused severe damage to the soil, including reduced fertility and disruption of microbial communities. Although \u003cem\u003eTrichoderma\u003c/em\u003e treatment has been shown to promote plant growth and improve soil quality, its effects on the growth promotion of pecan and the impact on soil microbial communities and physicochemical properties remained unclear.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eIn this study, we investigated the impact of \u003cem\u003eT. asperellum\u003c/em\u003e TCS007 spore suspension and its fermented crude extract on the growth and development of pecan seedlings. We also explored the effects of TCS007 treatment on the nutrients, enzyme activities, and microbial diversity in the rhizosphere soil of pecan seedlings during their three main growth stages.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eTreatment with TCS007 spore suspension or crude extract promoted the growth of pecan seedlings, with significantly higher levels of leaf hormones and defense enzyme activity compared to the control (CK). Moreover, the content of soil organic matter and ammonium nitrogen, as well as the activity of soil enzymes such as catalase and urease, were all significantly higher than CK after treatment, and the soil pH shifted from slightly acidic to slightly alkaline. The results indicated that TCS007 treatment significantly increased the richness of beneficial fungi and bacteria in the soil.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThe results demonstrated that TCS007 treatment significantly promoted the growth of pecan plants, increased enzyme activity and nutrient content in the soil, and improved the soil micro-ecological environment.\u003c/p\u003e","manuscriptTitle":"Exploring the potential of Trichoderma asperellum TCS007 on growth promotion of pecan seedlings as well as rhizosphere soil nutrients and microbial community","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-11 04:53:00","doi":"10.21203/rs.3.rs-5756661/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Accept","date":"2025-04-22T06:37:18+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2025-04-10T13:58:17+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-04-10T08:59:49+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-04-08T10:56:50+00:00","index":"","fulltext":""},{"type":"submitted","content":"Plant and Soil","date":"2025-04-07T22:10:10+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"plant-and-soil","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"plso","sideBox":"Learn more about [Plant and Soil](https://www.springer.com/journal/11104)","snPcode":"11104","submissionUrl":"https://submission.nature.com/new-submission/11104/3","title":"Plant and Soil","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"768d80ab-02d6-41e5-807e-fbd407529d49","owner":[],"postedDate":"April 11th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-05-19T16:02:29+00:00","versionOfRecord":{"articleIdentity":"rs-5756661","link":"https://doi.org/10.1007/s11104-025-07500-9","journal":{"identity":"plant-and-soil","isVorOnly":false,"title":"Plant and Soil"},"publishedOn":"2025-05-14 15:58:01","publishedOnDateReadable":"May 14th, 2025"},"versionCreatedAt":"2025-04-11 04:53:00","video":"","vorDoi":"10.1007/s11104-025-07500-9","vorDoiUrl":"https://doi.org/10.1007/s11104-025-07500-9","workflowStages":[]},"version":"v1","identity":"rs-5756661","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5756661","identity":"rs-5756661","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}