Trichoderma reesei endoglucanase ii

Abstract

The present application relates to a strain of sphaerostilbella pini YNF2404 and application thereof, and belongs to the technical field of biology, the preservation number of the strain is CCTCC NO: M 2026290, and the strain has the functions of decomposing organic / inorganic phosphorus, nitrogen fixation, promoting crop growth and preventing and treating banana wilt.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, and more specifically, relates to a strain of pine-loving bacterium YNF2404 and its applications. Background Technology

[0002] banana( Musa Bananas (Fusarium oxysporum) are a staple food for approximately 500 million people worldwide. my country is the world's second-largest banana producer, with banana production accounting for over 30% of the country's total tropical crop output. This disease is caused by Fusarium oxysporum caudata (specific strain). Fusarium oxysporum f. sp. cubense, Foc Caused by infection, this is a devastating soil-borne fungal disease that causes yellowing and wilting of the plant from the bottom up, longitudinal cracking of the pseudostem, and browning of the vascular bundles, ultimately leading to the death of the entire plant. The pathogen is mainly spread through rainwater, irrigation, seedlings, soil, and agricultural machinery and tools. It can survive in the soil for several years and accumulates year by year with continuous banana cropping, making it extremely difficult to control.

[0003] Since its first report in 1874, banana wilt disease remains unresolved and is a major bottleneck for the global banana industry. The disease can cause banana yield reductions of over 20%, and in severely affected fields, even total crop failure. In the late 1990s, banana wilt spread rapidly in my country. Taking Hainan, a major banana-producing region, as an example, the local planting area plummeted from a peak of 970,000 mu to less than 300,000 mu. In recent years, banana wilt has continued to occur at moderate to severe levels, with incidence rates exceeding 20% ​​in several producing areas, seriously hindering the healthy development of the banana industry.

[0004] Currently, the main methods for controlling banana wilt include breeding resistant varieties, chemical control, agricultural cultivation management, and biological control. Breeding resistant varieties is widely recognized as the fundamental solution, and my country has successively bred high-quality, high-yield wilt-resistant varieties such as "Baodao Banana," "Guijiao No. 9," "Zhongjiao No. 8," and "Zhongre No. 1." However, the breeding cycle for resistant varieties is long, and some varieties still have limitations in agronomic traits, quality, or regional adaptability. While chemical pesticides, as the main means of controlling plant fungal diseases, have achieved certain effects, their widespread use has led to serious soil pollution and increasingly prominent problems of pathogen resistance. Cultivation management measures (such as crop rotation and soil conditioning) are difficult and costly to implement, and cannot fundamentally eliminate pathogens in the soil. Therefore, exploring and applying new biological control resources and developing efficient, safe, and green control methods has become a major strategic need for promoting the sustainable development of the banana industry.

[0005] Biocontrol using functional microorganisms is an important component of the integrated management system for banana wilt disease. Numerous studies have shown that a large number of microbial strains with antagonistic activity against the causal agent of banana wilt exist in nature, including bacteria, fungi, and actinomycetes, primarily Bacillus, Trichoderma, and Streptomyces. In recent years, researchers have screened and obtained several biocontrol strains with good control effects against banana wilt. For example, Burkholderia cepacia GD1-1 showed a 72.5% inhibition rate against the mycelial growth of Fusarium oxysporum (Foc TR4), with a potted plant control efficacy of 55.6%, and also possessed the ability to solubilize potassium and phosphorus, produce proteases, and act as siderophores. Resveratrol GD3-16 achieved a 59.6% control efficacy against banana wilt, exhibited broad-spectrum antibacterial activity, and had a significant growth-promoting effect on banana plants. The combined use of Bacillus berreatus LMSY3Y-8 and hymexazol achieved a 60% control efficacy against banana wilt disease and significantly improved banana seedling height and root quality. Furthermore, the novel Streptomyces KHY26T demonstrated significant inhibitory effects on Foc TR4 and growth-promoting effects in greenhouse trials.

[0006] However, currently reported biocontrol strains for banana wilt generally suffer from limited functionality: most strains only possess the ability to antagonize the pathogen, lacking highly efficient strains that simultaneously exhibit multiple growth-promoting functions such as phosphorus solubilization and nitrogen fixation. Phosphorus in banana-growing soils exists primarily as insoluble phosphates, which plants cannot directly absorb and utilize, while nitrogen is an essential macronutrient for plant growth. If a biocontrol strain could be obtained that combines antagonistic activity against banana wilt pathogens with multiple functions including organic and inorganic phosphorus solubilization, nitrogen fixation, and growth promotion, it would be possible to achieve a synergistic effect of disease prevention and growth promotion, reducing the use of chemical fertilizers and pesticides, and providing new microbial resources for the green control of banana wilt.

[0007] genus Bassilis ( Talaromyces Bassicrystic fungi (BASF) are a class of highly adaptable saprophytic fungi widely distributed in soil, plant rhizosphere, and other environments. Some BASF promote plant mineral absorption, disease resistance, stress tolerance, and growth. Among them, *Bassicrys pineophilus* (*Bassicrys pineophilus*) Talaromyces pinophilus*Bacillus pineophilus* has been reported to possess multiple plant growth-promoting functions: it can secrete highly efficient cellulase to hydrolyze plant lignocellulose to increase soil humus, and it can also secrete organic acids to dissolve potassium feldspar and promote plant growth. Studies have shown that *Bacillus pineophilus* strain CA5 has the ability to produce cellulase, β-1,3-glucanase, and chitinase, and simultaneously possesses multiple growth-promoting functions such as inorganic phosphorus solubilization, organic phosphorus solubilization, potassium solubilization, nitrogen fixation, siderophore production, and IAA production. Other studies have shown that *Bacillus pineophilus* strain VRB36 possesses the ability to produce protease and amylase, as well as phosphorus solubilization, nitrogen fixation, and siderophore production, achieving a control efficiency of 56.92% against broad bean root rot, and significantly promoting the plant height, root length, and fresh / dry weight of broad bean seedlings. However, current research on the application of *Bacillus pineophilus* in the control of banana wilt disease is still very limited, and no *Bacillus pineophilus* strain specifically targeting banana wilt disease and possessing multiple functions including antagonism, organic phosphorus solubilization, inorganic phosphorus solubilization, nitrogen fixation, and growth promotion has been systematically reported and applied.

[0008] Therefore, screening and developing a new strain of *Pseudomonas pineophilus* that combines highly efficient antagonistic activity against *Fusarium wilt* of banana with multiple functions such as desiccation of organic and inorganic phosphorus, nitrogen fixation, and growth promotion, and applying it to the biological control of *Fusarium wilt* of banana, is of great significance for enriching the biocontrol microbial resources for *Fusarium wilt* of banana, reducing the use of chemical pesticides and fertilizers, and promoting the green and sustainable development of the banana industry. Summary of the Invention

[0009] In view of the shortcomings of the prior art, the present invention provides a strain of pine-loving basketbacter YNF2404 and its applications.

[0010] To achieve the above-mentioned objectives, the present invention provides the following technical solution: The first aspect of this invention provides a strain of *Pinocybe pineophilus* (… Talaromyces pinophilus The strain YNF2404 has the accession number CCTCC NO: M 2026290.

[0011] A second aspect of the present invention provides a microbial agent, wherein the active ingredient of the microbial agent is the *Pseudomonas aeruginosa* described in the first aspect.

[0012] The third aspect of this invention provides the application of the strain of the first aspect and the inoculant of the second aspect in the decomposition of inorganic / organic phosphorus, nitrogen fixation, promotion of crop growth, and control of banana wilt disease.

[0013] Furthermore, the pathogen causing banana wilt is *Fusarium oxysporum* var. *cubicans*. Fusarium oxysporum f. sp. cubense ( Foc )]4th physiological race.

[0014] Beneficial effects: I. The *Pseudomonas aeruginosa* strain provided by this invention is isolated from banana roots and has a better colonization effect compared to engineered strains; II. The *Pine-loving Basilella* provided by this invention has highly efficient antagonistic activity against banana wilt pathogens and functions of detoxifying organic and inorganic phosphorus, fixing nitrogen, and promoting growth. Attached Figure Description

[0015] Figure 1 This is the phylogenetic tree of *Pseudomonas pineae* in this invention; Figure 2 This is a diagram illustrating the antibacterial effect of *Pseudomonas aeruginosa* on *Fusarium wilt* of banana in this invention. In the diagram: the left image shows the antibacterial effect of *Pseudomonas aeruginosa* on *Fusarium wilt* of banana, and the right image shows the control (CK). Figure 3 A photograph of a plate-mounted qualitative strain of *Pseudomonas aeruginosa* with functions of lysing / degrading inorganic phosphorus and fixing nitrogen, as described in this invention. Figure 4 The control effect of the fermentation broth of *Pseudomonas aeruginosa* on TR4 is shown in the figure. In the figure, a represents YNF2404+TR4 and b represents CK+TR4. Figure 5 This is a diagram showing the growth-promoting effect of *Bacillus pineophilus* (YNF24041) on banana plants in this invention. In the diagram, a represents YNF2404 and b represents CK.

[0016] Biological Preservation The present invention provides *Pseudomonas aeruginosa* ( Talaromyces pinophilus YNF2404 is categorized as follows: Talaromyces pinophilus YNF2404 was deposited on January 29, 2026, at the China Center for Type Culture Collection (CCTCC), accession number CCTCC NO: M 2026290, located at Wuhan University, China. Detailed Implementation

[0017] The present invention will be described in detail below with reference to the embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0018] 1. The present invention provides a strain of *Pinocybean pineophilus* (…). Talaromyces pinophilus YNF2404 was deposited on February 5, 2026 at the China Center for Type Culture Collection (CCTCC), with accession number CCTCC NO: M 2026290. The address of the collection center is Wuhan University, China.

[0019] The morphological characteristics are: rapid growth on PDA plate medium, dense white hyphae, vigorous growth, and pale yellow colony reverse side.

[0020] 2. Talarophila piniphilum ( Talaromyces pinophilus Acquisition, identification and cultivation of ) 2.1 Source of Materials The fungus *Pine-loving Bassula* with accession number CCTCC NO: M 2026290 was isolated from the root system of banana plants infected with banana wilt in Jinping County, Honghe Prefecture, Yunnan Province.

[0021] 2.2 Culture medium Bengal Red Medium: 5g peptone, 1g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 10g glucose, 0.1g chloramphenicol, 0.033g Bengal Red, 20g agar, 1000 mL distilled water, pH 7.0.

[0022] PDA medium: 15g glucose, 15g agar, 200g peeled potato, 1000mL distilled water, pH 7.0. The liquid medium (PDB medium) is prepared without agar.

[0023] 2.3 Separation Method Sample disinfection: After rinsing the banana plant roots with sterile water, disinfect the surface with 75% ethanol for 60 seconds, and finally rinse with sterile water and air dry. The percentages mentioned are mass fractions.

[0024] Isolation and Culture: After disinfection of banana plant roots, the roots were cut into small pieces under aseptic conditions using a sterile knife as isolation material. The small pieces of banana roots were ground into a homogenate in a sterilized mortar. The pathogens were isolated by streak plating and cultured on Bengal Red agar. The culture was kept at 28°C for 3 days. Single colonies were picked and purified three times on PDA plates. Single spores were cultured and stored for later use.

[0025] 2.4 Classification and Identification The culture characteristics and morphological features of the strain were observed, and ITS amplification and sequence analysis were performed.

[0026] The observation and identification of culture traits and morphological characteristics were carried out in accordance with the "Methods for Plant Disease Research".

[0027] Molecular identification: Antagonistic bacterial DNA was extracted using the TSINGKE Plant DNA Extraction Kit (Universal). Based on the ITS sequence, primers ITS4 / ITS5 (5'-TCCCGTAGGTGAACCTGCGG-3' / 5'-TCCTCCGCTTATTGATATGC-3') were selected for PCR amplification. The primer sequences were synthesized by Beijing Qingke Xinyue Biotechnology Co., Ltd. The PCR products were sent to Beijing Qingke Xinyue Biotechnology Co., Ltd. for sequencing, and the sequencing results of the PCR amplification products were compared with BLAST on the NCBI website.

[0028] Based on culture characteristics, morphological observation, and analysis of the ITS sequence (SEQ ID NO.1) and phylogenetic tree, this strain was identified as *Pinococcus pyogenes*. Talaromyces pinophilus Its code name is YNF2404.

[0029] SEQ ID NO.1: .

[0030] 2.5 Cultivation and Preservation Methods Short-term culture and preservation: Inoculate on PDA solid slant medium, and after the bacteria have grown sufficiently, wrap the cotton plug part with oil paper and store in a refrigerator at 2-8℃.

[0031] Long-term culture and preservation: The cells were preserved in an ultra-low temperature freezer using the glycerol preservation method.

[0032] 3. Talarophila piniphilum ( Talaromyces pinophilus Test on the antibacterial activity of YNF2404 against Fusarium wilt of banana Fusarium wilt of bananas, race 4 (abbreviated) Foc4) As indicator bacteria, conventional confrontation culture methods were used to culture *Pseudomonas aeruginosa* (…). Talaromyces pinophilus The antibacterial activity of strain YNF2404 was determined. The pathogen causing banana wilt is *Fusarium oxysporum* var. *cubicans*. Fusarium oxysporum f. sp. cubense ( Foc strain No. 4 Foc 15-1, isolated and preserved by the banana research team of the Institute of Agricultural Environment and Resources, Yunnan Academy of Agricultural Sciences. The culture medium for the pathogen of banana wilt disease was PDA medium.

[0033] The inhibition rate of the strains was determined using the plate confrontation method. The pathogen of banana wilt was transferred to PDA plates and incubated at 28℃ for 7 days. A 5mm diameter mycelial cake was then punched along the edge of the colony using a sterilized punch and inoculated into the center of the PDA plate. Finally, *Pseudomonas aeruginosa* (a type of fungus) was picked up using an inoculation needle. Talaromyces pinophilus The inoculation was performed at 25 mm from the center of the PDA plate, with 4 points symmetrically inoculated per plate. Each treatment was repeated 3 times. PDA plates inoculated only with the tested pathogen served as a control. The antibacterial effect was observed after 7 days of incubation at 28 ℃. The pathogen of banana wilt was determined using the cross-hatching method. Foc 15-1) diameter (e.g. Figure 2 (As shown).

[0034] Inhibition rate (%) = (Coronavirus colony diameter of control pathogen - Coronavirus colony diameter of treatment pathogen) / Control diameter × 100.

[0035] Experimental data: The colony diameters of the pathogens in the three treatments were 2.62 cm, 2.61 cm, and 2.63 cm, respectively; the colony diameters of the pathogens in the three control treatments were 9.00 cm, 9.00 cm, and 9.00 cm, respectively. The inhibition rates of the three replicates were 70.89%, 71.00%, and 70.78%, respectively. The average inhibition rate of *Pseudomonas aeruginosa* against the pathogen of banana wilt was 70.89%, indicating a strong inhibitory effect on the growth of the pathogen.

[0036] 4. Determination of nitrogen-fixing and phosphorus-solubilizing activities of antagonistic bacteria of *Pseudomonas aeruginosa* Phosphate solubilization capacity determination Organic phosphorus solid medium: The organic phosphorus solid medium is the same as the inorganic phosphorus solid medium, but 5.00 g of tricalcium phosphate is replaced with 0.30 g of lecithin.

[0037] Inorganic phosphorus solid culture medium: glucose 10.0 g, (NH4)2SO4 0.5 g, NaCl 0.3 g, KCl 0.3 g, MgSO4•7H2O 0.3 g, FeSO4•7H2O 0.03 g, MnSO4•4H2O 0.03 g, Ca3(PO4)2 10 g, agar 18 g, distilled water 1000 ml, pH 7.0-7.5.

[0038] One colony of *Pseudomonas pineophilus* was inoculated onto organic and inorganic phosphorus solid agar plates, with three replicates per plate. The plates were incubated at 30 °C for 7 days. After 7 days, the presence of a clear zone around the colony was observed. If a clear zone was observed, it indicated that the selected strain had phosphorus solubilizing ability.

[0039] Nitrogen fixation capacity determination Ashby medium: 0.2 g potassium dihydrogen phosphate, 0.2 g magnesium sulfate, 0.2 g sodium chloride, 5.0 g calcium carbonate, 10.0 g mannitol, 0.1 g calcium sulfate, 18.0 g agar, 1000 mL distilled water, pH 6.8-7.0.

[0040] The *Pseudomonas pineophilus* strain was inoculated onto Asbestos nitrogen-free medium plates, with one colony per plate and three replicates. The plates were incubated at 30 °C for 7 days. After 7 days, the presence of a clear zone around the colony was observed. If a clear zone was observed, it indicated that the selected strain had nitrogen-fixing ability.

[0041] Experimental data: The diameters (D) of the three phosphorus-solubilizing zones for inorganic phosphorus solubilization activity were 2.60, 2.55, and 2.60, respectively; the colony diameters (d) were 1.60, 1.65, and 1.70, respectively; and the D / d ratios were 1.63, 1.55, and 1.53, respectively, with an average of 1.57.

[0042] The diameters (D) of the three solubilization zones for organophosphate activity were 2.30, 2.20, and 2.30, respectively; the colony diameters (d) were 1.50, 1.50, and 1.55, respectively; and the D / d ratios were 1.53, 1.47, and 1.48, respectively, with an average of 1.49.

[0043] The diameters (D) of the three nitrogen-fixing zones were 1.45, 1.40, and 1.45, respectively; the colony diameters (d) were 1.25, 1.20, and 1.30, respectively; and the D / d ratios were 1.16, 1.17, and 1.12, respectively, with an average of 1.15.

[0044] Qualitative plate analysis showed that this strain of *Pseudomonas pineophilus* (YNF2404) possesses activities involving the solubilization of inorganic and organic phosphorus, as well as nitrogen fixation (see attached). Figure 3(As shown in the image). This can provide a certain reference for the application of highly efficient phosphorus-solubilizing and nitrogen-fixing microorganisms and their microbial fertilizers. 5. Determination of the control efficacy and growth-promoting effect of *Pseudomonas pinephila* on potted banana wilt disease. Preparation of experimental banana seedlings: In a plastic greenhouse, tissue-cultured Brazilian banana seedlings (tissue-cultured seedlings from the Banana Research Laboratory of the Institute of Agricultural Environment and Resources, Yunnan Academy of Agricultural Sciences) were washed of the culture medium from their roots and transplanted into seedling bags. After the seedlings had grown 3-4 leaves (approximately one month), they were transplanted into plastic pots with a diameter of 11cm and a height of 12cm using vermiculite as the substrate. After transplanting, the seedlings were frequently watered to maintain moisture. Fertilizer was applied weekly (2g of compound fertilizer per plant dissolved in water, with a mass fraction of N-P2O5-K2O of 15-15-15) 1-2 times. Once the seedlings had grown 4-5 leaves (approximately one month), they were ready for use.

[0045] Preparation of the fermentation broth of the tested strain *Pseudomonas pineophilus* in this invention: The cryopreserved strain YNF2404 was streaked onto a solid PDA medium plate and activated in a 28 ℃ incubator for 5 days. The activated antagonistic bacteria were then inoculated into liquid PDA medium and cultured at 28 ℃ and 250 r / min for 7 days. The culture broth was filtered through four layers of sterile gauze to obtain the fermentation broth. A 1×10⁻⁶ concentration was prepared using sterile water. 8 Prepare antagonistic bacterial fermentation broth at cfu / mL.

[0046] Preparation of spore suspension of *Fusarium wilt* spores: *Fusarium wilt* spores were streaked onto solid PDA agar plates and activated in a 28°C incubator for 5 days. The activated *Fusarium wilt* mycelial cakes were then inoculated into liquid PDA agar and cultured at 28°C and 250 rpm for 7 days. The culture medium was filtered through four layers of sterile gauze to obtain a spore suspension. This suspension was then diluted with sterile water to a concentration of 1×10⁻⁶. 6 Prepare a solution of banana wilt spores at cfu / mL.

[0047] Banana Fusarium wilt control experiment in pots Control test method: The fermentation broth of the strain of banana wilt resistant to the present invention (fermentation broth of *Pseudomonas aeruginosa*, concentration 1×10⁻⁶) was used. 6 The solution (50 mL CFU / mL) was applied to the roots of the potted banana plants with uniform growth, while the control plants were treated with 50 mL PDB liquid culture solution per plant. After 7 days, the roots of the potted banana plants were treated to prevent root damage, and a solution of 1×10⁻⁶ CFU / mL PDB liquid culture solution was applied to the roots. 6 Apply 50 mL of cfu / mL banana wilt spore solution to the roots of potted banana plants, with a control of applying PDB liquid culture solution.

[0048] Control and investigation methods: Disease index was investigated 45 days after TR4 inoculation, examining both external symptoms (leaves) and internal symptoms (coronavirus tubers) for each treatment. Disease severity was determined based on the degree of browning in the longitudinal sections of banana leaves and corms, and the disease index and control effect were calculated. The disease index grading criteria are shown in Figure 1.

[0049] Table 1 Grading Standards for Banana Fusarium Wilt Disease

[0050] The pot experiment included two treatments: A1: irrigation with PDB liquid culture medium + Fusarium wilt spore liquid of banana (CK+TR4); A2: irrigation with fermentation broth of *Pseudomonas aeruginosa* + Fusarium wilt spore liquid of banana (YNF2404+TR4). Disease incidence was investigated for each treatment, disease index was calculated, and control efficacy was determined. The treatment results are shown in Table 2.

[0051] Table 2 Treatments in pot experiments

[0052] Potted plant control effect Through pot experiment, 45 days after inoculation with TR4 spore solution, as... Figure 4 As shown, the leaves of banana plants treated with only TR4 withered and turned yellow, with fewer leaves and shorter plants; while the leaves of banana plants treated with *Bacillus pineophilus* fermentation broth remained mostly healthy, with more leaves and better plant growth. Furthermore, after dissecting the corms, it was observed that the corms inoculated with the antagonistic bacteria were almost completely unaffected by TR4 mycelia, with only a few corms showing slight browning, while the corms inoculated with only TR4 turned brown and showed more brown mycelia.

[0053] 45 days after inoculation with TR4, the disease index of the bulb treated with *Bacillus pineophilus* fermentation broth was investigated, and the control effect was calculated. The results are shown in Table 3. The bulb disease index of the treatment treated with *Bacillus pineophilus* fermentation broth was significantly different from that of the control. The bulb disease index of the treatment treated with *Bacillus pineophilus* fermentation broth (YNF2404+TR4) was 17.50, and the leaf disease index was 26.67. The bulb disease index of the control was 47.67, and the leaf disease index was 60.83.

[0054] The control efficacy of irrigating with *Bacillus pineophilus* fermentation broth (YNF2404+TR4) on bulbs was 62.61%, and the control efficacy on leaves was 56.26%. The strain *Bacillus pineophilus* (YNF2404) showed good control effect against banana wilt.

[0055] Table 3. Control efficacy of *Pseudomonas pineophilus* (YNF2404) against banana wilt disease.

[0056] *: P-value less than 0.05 indicates a significant difference; **: P-value less than 0.01 indicates a significant difference; ***: P-value less than 0.001 indicates a very significant difference.

[0057] Plant growth promotion experiment Plant growth promotion test method: The fermentation broth of the strain of the present invention resistant to banana wilt (Fungiella pinephila fermentation broth: concentration 1×10⁻⁶) was used. 6 The solution (cfu / mL) was applied to the roots of potted banana plants with uniform growth, with 50 mL applied to each plant. The control group was treated with 50 mL of PDB liquid culture solution per plant.

[0058] Plant growth promotion survey method: On the day of irrigation with *Bacillus pineophilus* fermentation liquid (0 dpi) and 45 days after irrigation with *Bacillus pineophilus* fermentation liquid (45 dpi), bioinformatics indicators such as plant height, pseudostem diameter, number of leaves, leaf length, and leaf width of all treated banana plants were measured and recorded. In addition, bioinformatics indicators such as fresh weight of aboveground and underground parts of the plants were measured 45 days after irrigation with *Bacillus pineophilus* fermentation liquid.

[0059] Measurement and recording methods: Plant height: Measure the distance from the ground to the intersection of the petioles of the two top leaves; Leaf length: Measure the length from the tip to the tip of the first leaf at the top of the plant; Leaf width: Measure the width at the widest point of the first leaf at the top of the plant; Pseudostem diameter: Measure the diameter of the base of the pseudostem about 1 cm above the ground using calipers; Number of leaves: Record the number of green leaves per plant; Above-ground fresh weight: The weight of the banana plant above the base of the pseudostem about 1 cm above the ground; Below-ground fresh weight: The weight of the banana plant below the base of the pseudostem about 1 cm above the ground.

[0060] Table 3 shows the bioinformatics indicators of banana plants treated with *Bacillus pineophilus* fermentation broth on the same day (0 dpi) (0 dpi CK and 0 dpi YNF2404). There were no significant differences in the five bioinformatics indicators between the two treatments.

[0061] The plant growth promotion experiment included two treatments: B1: irrigation with PDB liquid culture medium, serving as the control (CK); B2: irrigation with *Pseudomonas pineophilus* fermentation broth (YNF2404). The treatment details are shown in Table 2.

[0062] Forty-five days after irrigating with *Bacillus pineophilus* fermentation broth, bioinformatics indicators of the treated banana plants were measured. Specific results are shown in Table 4 and... Figure 5As shown, the plant height, leaf length, leaf width, stem diameter, and number of leaves treated with *Bacillus pineophilus* fermentation broth (45 dpi YNF2404) were significantly different from the control (45 dpi CK), and the aboveground and underground fresh weights were significantly higher than the control. The results of this study indicate that the *Bacillus pineophilus* fermentation broth (45 dpi YNF2404) treatment promoted plant height, leaf length, leaf width, number of leaves, stem diameter, and aboveground and underground fresh weights compared to the control, with some indicators showing significant increases. This suggests that *Bacillus pineophilus* fermentation broth (YNF2404) has a growth-promoting effect on banana plants.

[0063] Table 4. Growth-promoting effects of *Bacillus pineophilus* (YNF2404) on banana plants.

[0064] *: P-value less than 0.05 indicates a significant difference; **: P-value less than 0.01 indicates a significant difference; ***: P-value less than 0.001 indicates a very significant difference.

[0065] This strain is an endogenous antagonistic functional bacterium with disease prevention, growth promotion, phosphorus solubilization, and nitrogen fixation effects. It can provide excellent strain resources in the fields of biological growth promotion and disease resistance in agriculture, and can lay the foundation for the development of efficient phosphorus solubilizing and nitrogen-fixing microorganisms for banana wilt disease and their microbial fertilizers.

[0066] The above description is merely a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the concept described herein through the above teachings or related technologies or knowledge. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention should be within the protection scope of the appended claims.

Claims

1. A strain of Talaromyces piniphilum ( Talaromyces pinophilus YNF2404, characterized in that: The preservation number of the *Pinococcus pyogenes* is: CCTCC NO: M 2026290, and its classification name is: Talaromyces pinophilus YNF2404.

2. A microbial agent, wherein the active ingredient of the microbial agent is the *Pseudomonas aeruginosa* as described in claim 1.

3. The application of the *Pseudomonas aeruginosa* as described in claim 1 or the inoculum as described in claim 2 in the decomposition of inorganic / organic phosphorus, nitrogen fixation, promotion of crop growth, and control of banana wilt disease.

4. The application according to claim 3, characterized in that: The pathogen causing banana wilt is *Fusarium oxysporum* var. *cubicans*. Fusarium oxysporum f. sp. cubense ( Foc )]4th physiological race.

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