A Trichoderma XH5 hybrid system, its construction method and application

Abstract

This invention relates to the field of water remediation technology, specifically disclosing a Trichoderma XH5 fungus, a hybrid system, its construction method, and its applications. The Trichoderma XH5 fungus was deposited at the China Center for Type Culture Collection on September 29, 2025, with accession number CCTCC: M 20252142, and is classified as follows: Eastern Trichoderma XH5. The Trichoderma XH5 provided by this invention has the ability to denitrify, specifically by improving the removal rates of nitrates, total nitrogen, and nitrites.

Description

Technical Field

[0001] This invention relates to the field of water remediation technology, specifically to a Trichoderma XH5, a hybrid system, its construction method, and its application. Background Technology

[0002] Groundwater is widely used as a drinking water source due to its stable reserves and excellent quality. However, due to excessive application of nitrogen fertilizers in agriculture, sewage leakage, and industrial emissions, nitrates have become one of the most widely distributed and highest-concentrated inorganic pollutants in groundwater. The high water solubility and low soil adsorption of nitrates make them easily diffused through seepage, leading to widespread nitrate exceedances in groundwater. Long-term intake of high nitrate levels can cause methemoglobinemia in infants, thyroid dysfunction, and potential carcinogenic risks, seriously threatening public health. Current main technologies for removing nitrates from water include physicochemical and biological methods. Physicochemical methods, including ion exchange, reverse osmosis, and electrodialysis, have drawbacks such as high energy consumption and expensive operating costs. In contrast, biological denitrification is preferred due to its low energy consumption, thorough denitrification, and lack of secondary pollution.

[0003] Therefore, it is necessary to develop a microorganism for denitrification. Summary of the Invention

[0004] To develop a microorganism for denitrification, this invention provides a Trichoderma XH5, a hybrid system, its construction method, and its applications. The Trichoderma XH5 provided by this invention possesses denitrification capabilities, specifically improving the removal rates of nitrates, total nitrogen, and nitrites.

[0005] This invention provides a Trichoderma ( Trichoderma orientale Trichoderma XH5, was deposited at the China Center for Type Culture Collection (CCTCC) on September 29, 2025, with accession number CCTCC: M 20252142, and is classified as follows: Trichoderma orientale XH5.

[0006] The Trichoderma XH5 provided by this invention has the ability to denitrify, specifically by improving the removal rates of nitrate, total nitrogen and nitrite.

[0007] The present invention also provides a hybrid system obtained by coupling the aforementioned Trichoderma XH5 with nanomaterials.

[0008] Furthermore, the nanomaterial is a multi-walled carbon nanotube material.

[0009] The method for constructing the hybrid system includes the following steps:

[0010] Trichoderma XH5 was inoculated into the culture medium and cultured until OD. 600The concentration was 0.1–0.15 to obtain XH5 bacterial solution; nanomaterials were added to XH5 bacterial solution and loaded at 300 r / min–400 r / min for 12 h–60 h to obtain hybrid system.

[0011] Furthermore, the final concentration of the multi-walled carbon nanotubes added to the XH5 bacterial solution is 1 g / L to 7 g / L.

[0012] The final concentration of the nanomaterial added to the XH5 bacterial solution was 5 g / L.

[0013] The present invention also provides an application of the aforementioned Trichoderma XH5 or the aforementioned hybrid system in water denitrification.

[0014] Furthermore, the application is to reduce the content of nitrate, total nitrogen and nitrite in water.

[0015] The present invention also provides a method for enhancing denitrification in water bodies, the method being based on the aforementioned hybrid system and comprising the following steps:

[0016] Multi-walled carbon nanotubes with a final concentration of 1 g / L to 7 g / L were combined with aerobic denitrifying fungi in the logarithmic growth phase. T. orientale XH5 bacterial culture was co-incubated on a magnetic stirrer to obtain... T. orientale XH5-multi-walled carbon nanotube hybrid system;

[0017] The hybrid system is then transferred to a water body to carry out a denitrification reaction, thereby reducing the levels of nitrate, total nitrogen, and nitrite in the water.

[0018] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0019] The Trichoderma XH5 isolated by this invention has the ability to denitrify, specifically by improving the removal rates of nitrate, total nitrogen and nitrite.

[0020] This invention combines multi-walled carbon nanotubes with aerobic denitrifying fungi. T. orientale XH5 physical coupling, forming T. orientale The XH5-MWCNTs hybrid system increases nitrate removal rate to 79%, fully utilizing the high conductivity and biocompatibility of MWCNTs to overcome the metabolic bottleneck of single strains and achieve rapid and efficient nitrogen removal. T. orientale The construction method of the XH5-MWCNTs hybrid system does not require the addition of an additional carbon source or complex processes, which can significantly reduce energy consumption and operating costs, and provides a feasible technical path for in-situ, efficient and green remediation of nitrate pollution in groundwater and landscape water bodies.

[0021] This invention discloses a nanomaterial-based aerobic denitrifying fungus. T. orientaleXH5 hybrid system for enhancing nitrogen removal in water bodies: Multi-walled carbon nanotubes at a final concentration of 5 g / L are combined with aerobic denitrifying fungi in the logarithmic growth phase. T. orientale XH5 bacterial culture was incubated on a magnetic stirrer at 350 rpm for 48 hours to obtain... T. orientale The XH5-multi-walled carbon nanotube composite hybrid system was centrifuged and transferred to a denitrification medium containing C / N=2 (N=10 mg / L) for denitrification. Compared with a single-strain system without multi-walled carbon nanotubes, the composite hybrid system of this invention improved nitrate removal rate from 62% to 79%, total nitrogen removal rate from 63% to 77%, and nitrite was completely removed within 30 hours, with no secondary accumulation of ammonia nitrogen. This invention utilizes the conductive network of multi-walled carbon nanotubes to promote direct electron transfer between bacteria and electron acceptors, effectively overcoming the metabolic bottleneck of single microorganisms. It eliminates the need for additional carbon sources and complex processes, offering advantages such as simple operation, low energy consumption, high stability, and no secondary pollution. It is suitable for in-situ efficient remediation of nitrate pollution in groundwater and landscape water bodies.

[0022] Information on the Preservation of Biological Materials

[0023] XH5, referred to as Trichoderma XH5 in this application, was deposited on September 29, 2025, at the China Center for Type Culture Collection (CCTCC), accession number CCTCC: M 20252142. The address of the depository is Wuhan University, No. 299 Bayi Road, Wuchang District, Wuhan, 430072, China. It is classified as follows: Trichoderma orientale XH5. Attached Figure Description

[0024] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0025] Figure 1 For the present invention T. orientale Phylogenetic tree and colony morphology of strain XH5;

[0026] In the figure, (a) is T. orientale Phylogenetic tree of strain XH5;

[0027] (b) is T. orientale Colony morphology of strain XH5.

[0028] Figure 2 For the present invention T. orientale XH5 and T. orientaleComparison of denitrification performance of XH5-MWCNTs hybrid systems;

[0029] In the figure, (a) is T. orientale The effect of strain XH5 on nitrogen concentration;

[0030] (b) is T. orientale The effect of strain XH5 on total nitrogen and nitrate nitrogen removal rates;

[0031] (c) is T. orientale The effect of XH5-MWCNTs hybrid system on nitrogen concentration;

[0032] (d) is T. orientale The effect of the XH5-MWCNTs hybrid system on the removal rates of total nitrogen and nitrate nitrogen.

[0033] Figure 3 The effect of different MWCNT concentrations on the present invention T. orientale The effect of XH5-MWCNTs hybrid system on denitrification performance;

[0034] In the figure, (a) shows the preparation when the final concentration of MWCNTs is 1 g / L. T. orientale Denitrification performance of the XH5-MWCNTs hybrid system;

[0035] (b) When the final concentration of MWCNTs is 3 g / L, the prepared T. orientale Denitrification performance of the XH5-MWCNTs hybrid system;

[0036] (c) When the final concentration of MWCNTs is 5 g / L, the prepared T. orientale Denitrification performance of the XH5-MWCNTs hybrid system;

[0037] (d) When the final concentration of MWCNTs is 7 g / L, the prepared T. orientale Denitrification performance of the XH5-MWCNTs hybrid system.

[0038] Figure 4 For the optimal load conditions of this invention T. orientale XH5-MWCNTs hybrid system and T. orientale The effects of XH5 on cell growth and DOC concentration;

[0039] In the figure, (a) is T. orientale The effects of XH5 on cell growth and DOC concentration;

[0040] (b) is T. orientale Effects of the XH5-MWCNTs hybrid system on cell growth and DOC concentration.

[0041] Figure 5 For the optimal load conditions of this invention T. orientaleXH5-MWCNTs hybrid system and T. orientale XH5 electrochemical impedance spectroscopy;

[0042] In the figure, (a) is T. orientale XH5 electrochemical impedance spectroscopy;

[0043] (b) is T. orientale Electrochemical impedance spectroscopy of the XH5-MWCNTs hybrid system.

[0044] Figure 6 For the optimal load conditions of this invention T. orientale XH5-MWCNTs hybrid system and T. orientale XH5 raw water denitrification performance;

[0045] In the figure, (a) is T. orientale XH5-MWCNTs hybrid system and T. orientale The effect of XH5 on the total nitrogen concentration in raw water;

[0046] (b) is T. orientale XH5-MWCNTs hybrid system and T. orientale The effect of XH5 on the nitrate nitrogen concentration in raw water;

[0047] (c) is T. orientale XH5-MWCNTs hybrid system and T. orientale The effect of XH5 on the concentration of nitrite nitrogen in raw water;

[0048] (d) is T. orientale XH5-MWCNTs hybrid system and T. orientale The effect of XH5 on the ammonia nitrogen concentration in raw water. Detailed Implementation

[0049] The specific embodiments of the present invention are described in detail below, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Unless otherwise specified, the experimental methods described in the embodiments of the present invention are conventional methods, and the materials and reagents used in the following embodiments are commercially available unless otherwise specified.

[0050] The products and formulas used in this invention are as follows:

[0051] Fungal solid culture medium: Weigh 31.6g of DRBC medium, add 1L of ultrapure water, and stir until completely dissolved. Autoclave at 121℃ for 30 minutes, and after the medium has cooled slightly to 50℃, pour the medium into sterile petri dishes on a sterile operating table for later use.

[0052] Denitrification liquid culture medium: KNO3 0.05 g / L, KH2PO4 1.5 g / L, glucose 0.072 g / L, MgSO4·7H2O 0.1 g / L, Na2HPO4·12H2O 5.0 g / L, trace element stock solution 2 mL; add the above components to ultrapure water and bring the volume to 1 L, stir until completely dissolved, then adjust the pH to 7.0, and autoclave at 121℃ for 30 minutes for later use.

[0053] The formula and preparation method of the trace element mother liquor are as follows: 4.4 mg ZnSO4, 100 mg ethylenediaminetetraacetic acid, 10.2 mg MnCl2·4H2O, 11 mg CaCl2, 10 mg FeSO4·7H2O, 3.2 mg CuSO4·5H2O, and 2.2 mg (NH4)6Mo7O 24 ·4H2O, 3.2 mg of CoCl2·6H2O; add the above components to ultrapure water and bring the volume to 1L, stir until completely dissolved, adjust the pH to 7.0, and autoclave at 121℃ for 30 minutes for later use.

[0054] PBS buffer: 2.865g Na2HPO4·12H2O, 0.312g NaH2PO4·2H2O; add the above components to ultrapure water to a final volume of 1L, stir until completely dissolved, then adjust the pH to 7.2, and autoclave at 121℃ for 30 minutes for later use.

[0055] 0.9% NaCl solution: Weigh 0.9g NaCl, add 100mL of ultrapure water, stir until completely dissolved, and autoclave at 121℃ for 30 minutes for later use.

[0056] The nanomaterial used in this invention is multi-walled carbon nanotubes, purchased from Shenzhen Suiheng Technology Co., Ltd., catalog number SH-GS-103. In this invention, it is referred to as MWCNTs, with a tube diameter of 3nm–15nm, a length of 15μm–30μm, and a purity >98%.

[0057] Example 1: A Trichoderma XH5 hybrid system, its construction method and application.

[0058] I. Experimental Methods

[0059] 1. Isolation and purification of bacterial strains

[0060] Fresh sediment and overlying water samples were collected at Fengqing Park in Xi'an City at a depth of 15 cm. The samples were collected using sterile sampling bottles that had undergone autoclaving. The sampling bottles were then immediately stored at low temperature after collection.

[0061] The collected water samples were mixed with denitrification liquid culture medium and placed in a 1000 mL Erlenmeyer flask. The mixture was then enriched and cultured in a shaking incubator at 30℃ and 130 r / min for 72 h.

[0062] The enriched bacterial suspension was serially diluted using a 10-fold dilution method and evenly spread onto fungal solid culture plates. The plates were inverted and placed in a 30°C dark incubator for 7 days until clearly morphologically distinct fungal colonies with well-defined edges were visible. After colony formation, single, well-grown, contamination-free colonies were picked using a flame-sterilized and cooled inoculation needle in a laminar flow hood and continuously streaked onto fungal solid culture plates for purification. This process was repeated until a purified strain was obtained. The selected strain was named XH5. DNA was extracted from the XH5 strain, and ITS sequencing technology was used to obtain genetic information and perform molecular identification. This strain was identified as... Trichoderma orientale abbreviated as T. orientale The colonies are yellow and diffuse. The phylogenetic tree and colony morphology of strain XH5 are as follows: Figure 1 As shown.

[0063] 2. T. orientale Construction of XH5-MWCNT hybrid system

[0064] Using a sterilized inoculation needle, pick the isolated and purified XH5 single strain and inoculate it into sterilized denitrification liquid medium. Incubate in a biochemical incubator shaker at 30°C and 120 rpm until OD500 is reached. 600 =0.15 to obtain bacterial solution.

[0065] In OD 600 Multi-walled carbon nanotubes were added to a bacterial culture with a concentration of 0.15 to achieve a final concentration of 5 g / L. A bacterial culture without added materials was used as a control group. Both cultures were placed on a magnetic stirrer and loaded at 350 rpm for 48 h to construct the desired bacterial culture. T. orientale XH5-MWCNTs hybrid system.

[0066] 3. Hybrid system and its effect on the denitrification performance of fungus XH5

[0067] Each T. orientale XH5 bacterial solution and T. orientale The XH5-MWCNTs hybrid system was centrifuged at 10000 r / min for 10 min, washed with PBS, and the supernatant was discarded. The resuspended components were then resuspended in 25 mL of 0.9% sodium chloride solution. Each resuspended component was transferred at a 10% (v / v) ratio to sterilized denitrification medium with a nitrogen concentration of 10 mg / L. The mixture was incubated in a biochemical shaking incubator at 30 °C and 120 rpm for 48 h. Samples were taken every 3 h, filtered through a pre-flamed 0.45 μm GF / F glass fiber filter, and the nitrate nitrogen (NO3) of each system was determined. --N, nitrite NO2 - -N, ammonia nitrogen NH4 + -N and total nitrogen (TN) concentrations.

[0068] like Figure 2 As shown, a single strain T. orientale XH5 removed approximately 62% of nitrates in the culture medium, and significant nitrite accumulation was observed between 12 and 30 hours, indicating the occurrence of aerobic denitrification. This was achieved with a final loading concentration of 5 g / L MWCNTs. T. orientale In the XH5-MWCNTs hybrid system, the removal rate of nitrate was increased to approximately 79%, and the total nitrogen removal rate was increased to approximately 77%. Furthermore, nitrite removal was rapidly completed within 30 hours, and no ammonia nitrogen accumulation was observed in the later stages. These results indicate that multi-walled carbon nanotubes can form low-resistance electron channels between the bacterial cells and electron acceptors, promoting bacterial attachment and interfacial electron transfer, achieving direct electron transfer and reducing energy loss, thereby improving aerobic denitrification efficiency.

[0069] 4. Fungi under multi-walled carbon nanotube loading conditions T. orientale Denitrification performance of XH5 at different material concentrations

[0070] To study the effects of different concentrations of MWCNTs on T. orientale To investigate the effect of the XH5-MWCNTs hybrid system on the nitrogen removal performance in water bodies, four final material concentrations were set at 1 g / L, 3 g / L, 5 g / L, and 7 g / L. The selected strains were first... T. orientale XH5 was cultured in a biochemical incubator shaker at 30℃ and 120 r / min until OD reached. 600 =0.15, obtained bacterial suspension, divided into four groups, and MWCNTs were added to make their final concentrations 1 g / L, 3 g / L, 5 g / L and 7 g / L respectively. The suspensions were loaded with a magnetic stirrer at 350 r / min for 48 h to obtain mixed suspensions of bacterial materials containing different final concentrations of MWCNTs. Each group of mixed suspensions was centrifuged, washed, and resuspended in 25 mL of 0.9% NaCl solution. The resuspended solution was added at a volume ratio of 10% to sterilized denitrification liquid medium with a nitrogen concentration of 10 mg / L. The mixture was incubated at 30℃ in a biochemical shaking incubator. Samples were taken every 3 h, filtered through a pre-flamed 0.45 μm GF / F glass fiber filter, and the total nitrogen (TN) and nitrate nitrogen (NO3) were measured. - -N.

[0071] Figure 3 show T. orientale The denitrification performance of XH5 was investigated at final MWCNT concentrations of 1 g / L, 3 g / L, 5 g / L, and 7 g / L, respectively. The effects of different final MWCNT concentrations on the nitrogen removal performance of the material were examined. T. orientaleThe effect of XH5 nitrogen reduction. When the final concentration of the material increased from 1 g / L to 7 g / L, the strain... T. orientale The nitrate removal rate of XH5 increased from 69.7% to 72.4%, reaching a maximum removal rate of 79% at 5 g / L. This indicates that the denitrification effect of the fungal-substrate mixture is enhanced with the increase of multi-walled carbon nanotube material concentration. However, when the concentration of multi-walled carbon nanotube material is too high, it may cause damage to the cell wall of the multi-walled carbon nanotube-perforated strain, thereby reducing the denitrification efficiency.

[0072] Taking into account both nitrate removal efficiency and nitrite accumulation, this invention determines 5 g / L as the optimal MWCNT loading concentration.

[0073] 5. Under optimal load conditions T. orientale XH5 and T. orientale Cell growth and DOC removal in the XH5-MWCNT hybrid system

[0074] fungi T. orientaele XH5 was inoculated into sterilized denitrification liquid medium, and multi-walled carbon nanotubes were added to a final concentration of 5 g / L. The mixture was then loaded for 48 h at 350 rpm using a magnetic stirrer. After centrifugation and washing, the mixture was resuspended in 25 mL of 0.9% NaCl solution. The resuspended solution was added at a 10% (v / v) ratio to sterilized denitrification liquid medium with a nitrogen concentration of 10 mg / L and a C / N ratio of 2. The mixture was incubated in a biochemical shaking incubator at 30 °C and 120 rpm. Samples were taken every 3 h, filtered through a pre-flamed 0.45 μm GF / F glass fiber filter, and the DOC of each system was measured. The OD of the unfiltered culture medium was measured. 600 .

[0075] like Figure 4 As shown, under the condition of no MWCNT loading, the adaptation period was 0h to 9h, during which cell growth was slow and DOC removal was also slow, decreasing from 20mg / L to 18.12mg / L; the logarithmic growth phase was 12h to 30h, during which the DOC content decreased from 17.89mg / L to 8.05mg / L, and decreased to the lowest value of 3.54mg / L by 48h.

[0076] exist T. orientale In the XH5-MWCNTs hybrid system, the adaptation period is from 0h to 9h, during which cell growth is slow and OD... 600 The concentration of DOC decreased from 0 to 0.026 mg / L, while DOC removal was also relatively slow, decreasing from 20 mg / L to 18.73 mg / L; the logarithmic growth phase of cells occurred from 12 to 33 hours. T. orientaleIn the XH5-MWCNTs hybrid system, the DOC concentration decreased from 17.07 mg / L to 6.04 mg / L, reaching a minimum of 2.24 mg / L at 48 h. The DOC content gradually decreased as denitrification proceeded, indicating that cell self-replication requires DOC, while simultaneously providing the necessary electron donors for denitrification.

[0077] 6. The effect of loading multi-walled carbon nanotubes on the electron transfer efficiency of hybrid systems

[0078] Will T. orientale XH5 was inoculated into sterilized denitrification liquid medium, and multi-walled carbon nanotubes were added to a final concentration of 5 g / L. The mixture was then loaded for 48 h at 350 rpm using a magnetic stirrer. After centrifugation, washing, and resuspending in 25 mL of 0.9% NaCl solution, a culture without multi-walled carbon nanotubes was also included. T orientale XH5 bacterial culture was used as the control group.

[0079] The resuspended solution was added at a volume ratio of 10% to sterilized denitrification liquid culture medium with a nitrogen concentration of 10 mg / L. A three-way bottle was used as the reactor. The reference electrode (saturated calomel electrode), the counter electrode (platinum sheet electrode), and the working electrode (glassy carbon electrode) were assembled in the three-way bottle, ensuring that the electrodes did not contact each other. The reaction apparatus was then constructed. The three-way bottle was placed on a magnetic stirrer and incubated at 200 rpm for 48 h. Electrochemical impedance spectroscopy was performed at 16 h, 32 h, and 48 h.

[0080] Electrochemical impedance spectroscopy: Connect the device to the electrochemical workstation, open the software, and after the open-circuit voltage stabilizes, select IMP-ACImpedance and set the parameters as follows: Init E: Open-circuit voltage value in V; High Frequency: 10 6 Hz; LowFrequency: 0.1Hz; Amplitude: 0.1V.

[0081] The impedance spectrum consists of two characteristic regions: the first is a semi-circular structure in the high-frequency region, which mainly corresponds to the charge transfer process at the electrode / electrolyte interface. The diameter of the semi-circle is directly related to the charge transfer resistance; the magnitude of the charge transfer resistance is mainly affected by the electrode reaction kinetics, the bioelectrochemical reaction rate within the system, and the charge transfer efficiency between the biomembrane and the electrode interface. The second is a linear structure in the low-frequency region, which corresponds to the diffusion-controlled process of reactants or products in the electrode reaction system. The charge transfer resistance is denoted as Rct.

[0082] like Figure 5 As shown in (a), in T. orientale In the XH5 system, the Rct values ​​in the early, middle, and late stages of the reaction showed a significant decreasing trend. This is because, as the culture time progressed, the fungi... T. orientaleThe metabolic activity of XH5 is significantly enhanced, and the fungus secretes a large amount of extracellular electron transport mediators, promoting electron transfer to the electrode. For example... Figure 5 (b), relative to T. orientale XH5 system, T. orientale The Rct of the XH5-MWCNTs hybrid system is relatively small, which indicates that the loading of multi-walled carbon nanotubes enables the redox-active molecules such as riboflavin and quinones secreted by the strain to synergistically form a conductive network with MWCNTs, thereby accelerating the transfer of electrons from the bacterial cell to the electrode and improving the overall electron transfer efficiency.

[0083] 7. Raw water verification

[0084] Water samples were collected from Qujiang Park, Xingqing Palace, and Changle Park in Xi'an City. The samples were designated as QJ, XQ, and CL, respectively. The sampling depth was 15 cm. The samples were collected using sterile sampling bottles that had undergone autoclaving. The sampling bottles were immediately stored at low temperature after collection.

[0085] Will T. orientale XH5 was inoculated into sterilized denitrification liquid medium, and multi-walled carbon nanotubes were added to a final concentration of 5 g / L. The mixture was then loaded for 48 h at 350 rpm using a magnetic stirrer. After centrifugation, washing, and resuspending in 25 mL of 0.9% NaCl solution, a culture without multi-walled carbon nanotubes was also included. T orientale XH5 bacterial culture was used as the control group.

[0086] The resuspension was added to 950 mL of raw water sample at a 5% (v / v) ratio, and cultured in a 1 L reactor for 5 days. An oxygenation pump was used to maintain the aerobic environment of the water. Samples were taken every day, filtered through a pre-burned 0.45 μm GF / F glass fiber filter, and the nitrate nitrogen and NO3 of each system were measured. - -N, nitrite NO2 - -N, ammonia nitrogen NH4 + -N and total nitrogen (TN) concentrations. QJ, XQ, and CL represent water bodies with low, medium, and high pollution levels, respectively, with TN concentrations of 2.09, 3.098, and 4.07 mg / L.

[0087] The results are as follows Figure 6 As shown, in the QJ raw water reaction system, T. orientale In the XH5 system, the TN concentration decreased from 2.09 mg / L to 1.02 mg / L; T. orientale In the XH5-MWCNTs hybrid system, the TN concentration decreased from 2.09 mg / L to 0.19 mg / L, essentially achieving TN removal. In the XQ raw water reaction system, T. orientale In the XH5 system, the TN removal rate was 64.26%; T. orientaleIn the XH5-MWCNTs hybrid system, the TN removal rate increased to 95.47%. In the CL raw water reaction system, it... T. orientale Compared to the XH5 system T. orientale The TN removal rate in the XH5-MWCNTs hybrid system increased from 54.29% to 91.66%. Furthermore, compared to... T. orientale XH5 system, T. orientale NO3 in the XH5-MWCNTs hybrid system - -N, NO2 - -N, NH4 + -N all decreased to varying degrees. These phenomena indicate that... T. orientale The XH5-MWCNTs hybrid system has the potential to purify water with varying degrees of micro-pollution.

[0088] Although preferred embodiments of the invention have been described, those skilled in the art, once they have learned the basic inventive concept, can make other changes and modifications to these embodiments.

[0089] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A type of Trichoderma Trichoderma orientale XH5, characterized in that, Trichoderma XH5 was deposited at the China Center for Type Culture Collection on September 29, 2025, with accession number CCTCC: M 20252142.

2. A hybrid system, characterized in that, The hybrid system is obtained by coupling Trichoderma XH5 as described in claim 1 with nanomaterials; the nanomaterials are multi-walled carbon nanotubes.

3. A method for constructing the hybrid system according to claim 2, characterized in that, Includes the following steps: Trichoderma XH5 was inoculated into the culture medium and cultured until OD. 600 The concentration was 0.1–0.15 to obtain XH5 bacterial solution; nanomaterials were added to XH5 bacterial solution and loaded at 300 r / min–400 r / min for 12 h–60 h to obtain hybrid system.

4. The method for constructing a hybrid system according to claim 3, characterized in that, The final concentration of the nanomaterial added to the XH5 bacterial solution is 1 g / L to 7 g / L.

5. The method for constructing a hybrid system according to claim 4, characterized in that, The final concentration of the nanomaterial added to the XH5 bacterial solution was 5 g / L.

6. The application of Trichoderma XH5 as described in claim 1 or the hybrid system as described in claim 2 in water denitrification.

7. The application according to claim 6, characterized in that, The application is to reduce the content of nitrate, total nitrogen and nitrite in water.

8. A method for enhancing nitrogen removal from water bodies, characterized in that, The method is based on the hybrid system described in claim 2 and includes the following steps: The hybrid system is then transferred to a water body to carry out a denitrification reaction, thereby reducing the levels of nitrate, total nitrogen, and nitrite in the water.

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