Pseudomonas ginseng and application thereof

Abstract

The invention discloses Pseudomonas ginseng and an application thereof. The invention provides Pseudomonas ginsengi, the strain number of the Pseudomonas ginsengi is L1012, and the preservation number of the Pseudomonas ginsengi in the General Microbiological Culture Collection Center of the China Committee for Culture Collection of Microorganisms is CGMCC No.32197. The strain L1012 has inhibitory activity on root rot pathogenic bacteria Fusarium equiseti, has good potential in biological prevention and control of root rot of medicinal plants (such as pseudo-ginseng, ginseng and bighead atractylodes rhizome), and can also be used for soil remediation, soil improvement and the like of planting fields of pseudo-ginseng, ginseng and bighead atractylodes rhizome.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to Pseudomonas ginseng and its applications. Background Technology

[0002] The genus *Pseudomonas* belongs to the domain Bacteria, phylum Pseudomonadota, class Gammaproteobacteria, order Pseudomonadales, and family Pseudomonadaceae. *Pseudomonas* exhibits extremely high species diversity and a wide ecological distribution; strains have been isolated from soil, water bodies, plant surfaces, and internal tissues. The type species of this genus is *Pseudomonas fluorescens*, with the type strain ATCC13525. T (https: / / lpsn.dsmz.de / genus / pseudomonas).

[0003] Currently, over 200 effective species of the genus *Pseudomonas* have been described, and they are widely used in agriculture, industry, and environmental remediation. For example, *Pseudomonas putida* is an important industrial microorganism (Qi X, Gao X, Wang X, Xu P. Harnessing *Pseudomonas putida* in bioelectrochemical systems. Trends Biotechnol. 2024 Jul; 42(7):877-894. doi:10.1016 / j.tibtech.2023.12.002. Epub 2024 Jan 5. PMID:38184440.), and has been extensively studied due to its metabolic diversity and ability to degrade toxic compounds; *Pseudomonas aeruginosa* has attracted much attention due to its multidrug resistance and its importance in hospital-acquired infections (Mielko KA, SJ, Milczewska J, Sands D M, P. Metabolomics studies of Pseudomonas aeruginosa. World J Microbiol Biotechnol. 2019 Nov 7; 35(11): 178. doi: 10.1007 / s11274-019-2739-1. PMID: 31701321; PMCID: PMC6838043.).

[0004] Pseudomonas strains possess a variety of biological activities and ecological functions. For example, Pseudomonas fluorescens can produce various antibiotics and siderophores, inhibiting the growth of plant pathogens, thus having important application value in biological control. J, Chauhan A, Ripp S, Layton AC, Kuncová G, Sayler GS. Pseudomonas fluorescens HK44: lessons learned from a model whole-cell bioreporter with a broad application history. Sensors (Basel). 2012; 12(2):1544-71. doi:10.3390 / s120201544. Epub 2012 Feb 6. PMID:22438725;PMCID:PMC3304127.). Pseudomonas stutzeri has been studied in agriculture for its nitrogen-fixing ability to improve soil fertility (Sun X, Xu Z, Xie J, Hesselberg-Thomsen V, Tan T, Zheng D, Strube ML, A, Shen Q, Zhang R, Kovács Baci l lus velezens i sstimulates resident rhizosphere Pseudomonasstutzeri for plant health through metabolic interactions.ISME J.2022Mar;16(3):774-787.doi:10.1038 / s41396-021-01125-3.Epub 2021Sep 30.PMID:34593997; PMCID:PMC8483172.). Studies have shown that *Pseudomonas* strains occupy an important position in the microbial communities of polluted soil and water bodies and play a key role in the degradation of pollutants (Palleroni NJ. The Pseudomonas story. Environ Microbiol. 2010 Jun; 12(6):1377-83. doi:10.1111 / j.1462-2920.2009.02041.x.PMID:20553550.). Therefore, *Pseudomonas* strains are a diverse and widely used group of microorganisms. Their importance in ecosystems and their potential applications in biotechnology make them a hot topic in microbiology research. Therefore, exploring and discovering new *Pseudomonas* resources using new microbiological research techniques has significant scientific value and application prospects. Summary of the Invention

[0005] This invention claims protection for Pseudomonas ginseng and its applications.

[0006] In a first aspect, the present invention claims protection for Pseudomonas ginsengi.

[0007] The present invention claims protection for *Pseudomonas ginsengi*, strain number L1012, which has accession number CGMCC No. 32197 at the China General Microbiological Culture Collection Center.

[0008] The claimed strain of *Pseudomonas ginsengi*, L1012, is a Gram-negative aerobic heterotrophic bacterium. On PYG solid medium, its colonies are pale yellow, moist, smooth, and have intact edges. The cells are motile, rod-shaped, and measure (0.6-0.8) × (1.1-1.2) μm. The growth tolerance range of strain L1012 is 10-37℃, 0-5% NaCl, and pH 6.0-8.0, with optimal growth conditions of 30℃, 0% NaCl, and pH 7.0. The strain is positive for oxidase and catalase tests, but negative for nitrate reduction and gelatin liquefaction tests. It can assimilate D-glucose, D-maltose, D-mannose, and D-fructose as its sole carbon source and produce acid.

[0009] Secondly, this invention claims protection for a culture.

[0010] The culture claimed in this invention is the culture of Pseudomonas ginsengi described in the first aspect above, which is a substance obtained by culturing Pseudomonas ginsengi described in the first aspect above in a bacterial culture medium.

[0011] The aforementioned culture contains the substance comprising the *Pseudomonas ginsengi* (the bacterial cell itself) and the fermentation broth of the *Pseudomonas ginsengi* or a dilution thereof.

[0012] In the above-mentioned cultures, the bacterial culture medium can be a solid culture medium or a liquid culture medium.

[0013] The term "culture" refers to any liquid or solid culture medium that has been artificially inoculated and cultivated, resulting in the growth of a microbial community. It is the product obtained through the growth and / or amplification of microorganisms; it can be a biologically pure culture of microorganisms, or it can contain a certain amount of culture medium, metabolites, or other components produced during the cultivation process. The term "culture" also includes passaged cultures obtained by subculturing microorganisms; these can be cultures of a single generation or mixtures of several generations. The term "metabolite" refers to the primary and / or secondary metabolites produced during the metabolism of microorganisms. Primary metabolism refers to the process by which microorganisms absorb various nutrients from the external environment and, through catabolism and anabolism, generate substances and energy to sustain life activities. The products of primary metabolism are called primary metabolites, such as monosaccharides or monosaccharide derivatives, nucleotides, vitamins, amino acids, fatty acids, and other monomers, as well as various macromolecular polymers composed of them, such as proteins, nucleic acids, polysaccharides, and lipids. Secondary metabolism refers to the process by which microorganisms, at a certain growth stage, use primary metabolites as precursors to synthesize substances that do not have a clearly defined function in the life activities of microorganisms. The products of secondary metabolism are mostly compounds with relatively complex molecular structures. Based on their functions, they can be classified into types such as antibiotics, hormones, alkaloids, and toxins.

[0014] In a specific embodiment of the present invention, the bacterial culture medium is specifically PYG culture medium or fermentation culture medium.

[0015] In one embodiment of the present invention, the solvent of the fermentation medium is water, and the solvent and concentration are as follows: glucose 5g / L, malt extract 10g / L, yeast extract 5g / L, cottonseed cake 10g / L, soluble starch 20g / L, potassium dihydrogen phosphate 0.5g / L, ammonium sulfate 5g / L, calcium carbonate 3g / L, sodium chloride 1g / L, pH 7.2.

[0016] Thirdly, the present invention claims protection for a fermentation broth or a dilution thereof obtained by inoculating the *Pseudomonas ginsengi* described in the first aspect above into a fermentation medium.

[0017] The term "fermentation broth" refers to a liquid fermentation system that has grown a microbial community after artificial inoculation and fermentation culture.

[0018] In one embodiment of the present invention, the solvent of the fermentation medium is water, and the solvent and concentration are as follows: glucose 5g / L, malt extract 10g / L, yeast extract 5g / L, cottonseed cake 10g / L, soluble starch 20g / L, potassium dihydrogen phosphate 0.5g / L, ammonium sulfate 5g / L, calcium carbonate 3g / L, sodium chloride 1g / L, pH 7.2.

[0019] In one embodiment of the present invention, the diluent is obtained by diluting the fermentation broth with sterile distilled water.

[0020] Fourthly, this invention claims protection for a microbial agent.

[0021] The microbial agent claimed in this invention contains *Pseudomonas ginsengi* as described in the first aspect above, the culture as described in the second aspect above, and / or the fermentation broth or its dilution as described in the third aspect above.

[0022] Furthermore, the fungal agent may be a fungal agent for preventing and controlling root rot and / or a fungal agent for inhibiting Fusarium equisetifolium.

[0023] In the microbial agent, the active ingredient includes or is the *Pseudomonas ginsengi* described in the first aspect above, the culture described in the second aspect above, and / or the fermentation broth or its dilution described in the third aspect above.

[0024] In the aforementioned microbial agent, in addition to the active ingredient, a carrier is also included. The carrier can be a commonly used and biologically inert carrier in the pesticide field. The carrier can be a solid or liquid carrier; the solid carrier can be a mineral material, plant material, or polymer compound; the mineral material can be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica, and diatomaceous earth; the plant material can be at least one of corn flour, soybean flour, and starch; the polymer compound can be polyvinyl alcohol and / or polyethylene glycol; the liquid carrier can be an organic solvent, vegetable oil, mineral oil, or water; the organic solvent can be decane and / or dodecane.

[0025] The above-mentioned microbial agents can be in various formulations, such as liquid, emulsion, suspension, powder, granules, wettable powder or water-dispersible granules.

[0026] Depending on the requirements, surfactants (such as Tween 20, Tween 80, etc.), binders, stabilizers (such as antioxidants), pH adjusters, etc. may also be added to the bacterial agent.

[0027] Fifthly, the present invention claims protection for the use of *Pseudomonas ginsengi* as described in the first aspect above, or the culture as described in the second aspect above, or the fermentation broth or its dilution as described in the third aspect above, or the inoculant as described in the fourth aspect above, in any of the following:

[0028] (A1) Inhibits Fusarium equisetifolium;

[0029] (A2) Prepare products for inhibiting Fusarium equisetifolium;

[0030] (A3) Prevention and control of root rot;

[0031] (A4) Prepare products for the prevention and control of root rot.

[0032] Furthermore, the root rot may be caused by Fusarium equisetifolium.

[0033] Furthermore, the root rot refers to root rot of medicinal plants. These medicinal plants may include Panax notoginseng, ginseng, or Atractylodes macrocephala, among others.

[0034] Sixthly, the present invention claims protection for the use of *Pseudomonas ginsengi* as described in the first aspect above, or the culture as described in the second aspect above, or the fermentation broth or its dilution as described in the third aspect above, or the inoculant as described in the fourth aspect above, in any of the following:

[0035] (B1) Soil remediation of medicinal plant cultivation sites;

[0036] (B2) Prepare products for soil remediation in medicinal plant cultivation sites;

[0037] (B3) Soil improvement in medicinal plant cultivation sites;

[0038] (B4) Prepare products for soil improvement in medicinal plant cultivation sites.

[0039] The soil remediation or soil improvement refers to inhibiting the activity of root rot pathogen (Fusarium equisetifolium) in the soil of medicinal plant planting sites.

[0040] Furthermore, the medicinal plant may be Panax notoginseng, ginseng, or Atractylodes macrocephala, etc.

[0041] Seventhly, the present invention claims protection for a product for inhibiting Fusarium equisetifolium.

[0042] The product for inhibiting Fusarium equisetifolium claimed in this invention contains, or is comprised of, the Pseudomonas ginsengi described in the first aspect above, the culture described in the second aspect above, the fermentation broth described in the third aspect above, or a diluted solution thereof, or the microbial agent described in the fourth aspect above.

[0043] The product may be an antibacterial agent.

[0044] Eighthly, the present invention claims protection for a product used to prevent and control root rot.

[0045] The product for preventing and controlling root rot claimed in this invention contains, or is comprised of, the *Pseudomonas ginsengi* as described in the first aspect above, the culture as described in the second aspect above, the fermentation broth or its dilution as described in the third aspect above, or the microbial agent as described in the fourth aspect above.

[0046] The product may be a biological agent.

[0047] Ninthly, the present invention claims a method for inhibiting Fusarium equisetifolium.

[0048] The method for inhibiting Fusarium equisetifolium claimed in this invention may include the following steps: treating the Fusarium equisetifolium or its host with Pseudomonas ginsengi as described in the first aspect above, or the culture as described in the second aspect above, or the fermentation broth or its dilution as described in the third aspect above, or the inoculant as described in the fourth aspect above.

[0049] In a tenth aspect, the present invention claims a method for preventing and controlling root rot.

[0050] The method for preventing and controlling root rot claimed in this invention may include the following steps: treating the plant or its growth substrate to be controlled with Pseudomonas ginsengi as described in the first aspect above, or the culture as described in the second aspect above, or the fermentation broth or its dilution as described in the third aspect above, or the inoculant as described in the fourth aspect above.

[0051] Eleventhly, the present invention claims the use of Pseudomonas ginsengi as described in the first aspect above in the preparation of the culture described in the second aspect above, the fermentation broth or its dilution described in the third aspect above, or the inoculum described in the fourth aspect above.

[0052] In one embodiment of the present invention, the *Fusarium equiseti* is specifically *Fusarium equiseti* CGMCC 3.15353.

[0053] Experiments have demonstrated that strain L1012 of this invention represents a novel species of the genus Pseudomonas, based on phenotypic and genotypic comparisons with existing Pseudomonas strains. According to its isolation from ginseng rhizosphere soil, this new species is named *Pseudomonas ginsengi*. Activity testing confirmed that strain L1012 exhibits inhibitory activity against *Fusarium equiseti*, the pathogen causing root rot. These results suggest that strain L1012 has good potential for the biocontrol of root rot in medicinal plants (such as *Panax notoginseng*, ginseng, and *Atractylodes macrocephala*), and can also be used for soil remediation and improvement in *Panax notoginseng*, ginseng, and *Atractylodes macrocephala* cultivation areas.

[0054] Preservation Instructions

[0055] Classification and nomenclature: Pseudomonas ginsengi;

[0056] Biological material of ginseng: L1012;

[0057] Preservation institution: China General Microbiological Culture Collection Center, China Committee on the Preservation and Management of Microbial Cultures;

[0058] The abbreviation for the depository institution is CGMCC.

[0059] Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing;

[0060] Deposit date: October 14, 2024;

[0061] Registered with the China National Collection Center (CGMCC) No. 32197. Attached Figure Description

[0062] Figure 1 The image shows the inhibitory activity of strain L1012 against Fusarium equiseti CGMCC 3.15353. In the image, a represents the original fermentation broth of strain L1012; b represents a 10-fold dilution of the fermentation broth of strain L1012; and c represents a 100-fold dilution of the fermentation broth of strain L1012.

[0063] Figure 2 The NJ phylogenetic tree is constructed based on the 16S rRNA gene sequences of L1012 and closely related strains of the genus Pseudomonas. Detailed Implementation

[0064] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0065] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0066] Example 1: Isolation and Identification of Strains L1012

[0067] I. Isolation of strain L1012

[0068] The strain L1012 of this invention was isolated from ginseng rhizosphere soil, and the sample was collected from a ginseng planting base in Chuxiong Yi Autonomous Prefecture, Yunnan Province. The specific isolation procedures are as follows:

[0069] Soil suspension preparation: 2g of soil sample was added to 18mL of sterile mixed solution containing 0.1% sodium pyrophosphate and 0.85% sodium chloride, and placed in a shaker at 28℃ and 180rpm for 40min to fully suspend the soil particles. The resulting soil suspension was then serially diluted to 10-1. -4 spare.

[0070] Isolation medium: 10 g / L soluble starch -1 dipotassium hydrogen phosphate 1 g·L -1 Magnesium sulfate heptahydrate 1 g·L -1 ammonium sulfate 2 g·L -1 Calcium carbonate 2g·L -1 Ferrous sulfate heptahydrate 0.001 g·L -1 0.001 g·L⁻¹ manganese chloride -1 Zinc sulfate 0.001 g·L -1 Sodium chloride 1 g·L -1 12.5 mL / L glycerol -1 Arginine 2g·L -1 15g / L agar -1 pH 7.5.

[0071] Isolation method: Spread the diluted soil suspension onto isolation medium plates and incubate at 30°C for 3 weeks. Select healthy single colonies and inoculate them onto PYG slant medium (formulation: 3 g / L peptone). -1 5g / L of yeast extract -1 10 g / L of glycerin -1 betaine 1.25 g·L-1 Sodium pyruvate 1.25 g·L -1 15g / L agar -1 The culture was cultured at pH 7 to obtain a pure culture, from which a strain numbered L1012 was isolated and purified. For subsequent research, the pure strain was preserved in liquid nitrogen and frozen at -80°C using 20% ​​(v / v) glycerol as a protectant.

[0072] II. Detection of physiological and biochemical characteristics of strain L1012

[0073] Strain L1012 was inoculated onto tryptone soybean broth medium (Soluble) at 28°C, and its morphology, physiological and biochemical properties, cellular fatty acid composition, and gene levels were investigated. The physiological and biochemical characteristics of the strain were tested using API 50CH and API ZYM assay kits and the corresponding procedures. Other physiological and biochemical characteristics of the strain, including Gram staining properties, oxygen requirement, catalase activity, and oxidase activity, were determined according to the *Handbook of Systematic Identification of Common Bacteria* (Dong Xiuzhu & Cai Miaoying. 2001. *Handbook of Systematic Identification of Common Bacteria*. Beijing: Science Press).

[0074] The identification results showed that strain L1012 was a Gram-negative aerobic heterotrophic bacterium. Colonies formed on PYG solid medium were pale yellow, moist, smooth, and with intact edges. The cells were motile, rod-shaped, and measured (0.6-0.8) × (1.1-1.2) μm. The growth tolerance range of strain L1012 was 10-37℃, 0-5% NaCl, and pH 6.0-8.0, with the optimal growth conditions being 30℃, 0% NaCl, and pH 7.0. Strain L1012 was positive for oxidase and catalase tests, but negative for nitrate reduction and gelatin liquefaction tests. It could assimilate D-glucose, D-maltose, D-mannose, and D-fructose as its sole carbon source and produce acid (Table 1).

[0075] Table 1. Physiological and biochemical test results of strain L1012

[0076]

[0077]

[0078]

[0079] Note: + indicates positive; - indicates negative.

[0080] III. Detection of cellular fatty acid components in strain L1012

[0081] The cellular fatty acid chemical composition of strain L1012 was detected by gas chromatography (Sasser M. Identification of bacteria by gas ghromatography of cellular fatty acids, MIDI Technical Note 101. Newark, DE:MIDI inc; 1990.).

[0082] The fatty acid composition of strain L1012 of this invention is shown in Table 2. The results show that the main fatty acid of strain L1012 is Sum Feature 3 (C... 16:1 ω7c / C 16:1 ω6c) and C 16:0 These components accounted for 47.7% and 31.9% of the total content, respectively, and these main components are consistent with the main fatty acid components of Pseudomonas strains.

[0083] Table 2. Fatty acid composition of strain L1012 cells

[0084]

[0085]

[0086] IV. Inhibitory activity test of the strain against Fusarium equiseti CGMCC 3.15353, the pathogen of root rot.

[0087] The inhibition zone method was used to test the inhibitory activity of strain L1012 against Fusarium equiseti CGMCC 3.15353. Specific test steps:

[0088] (1) Preparation of fermentation broth of strain L1012: strain L1012 was inoculated into fermentation medium (formula: glucose 5g, malt extract 10g, yeast extract 5g, cottonseed cake 10g, soluble starch 20g, potassium dihydrogen phosphate 0.5g, ammonium sulfate 5g, calcium carbonate 3g, sodium chloride 1g, deionized water 1L, pH 7.2), placed in a shaker, and cultured at 30℃ for 4 days at a speed of 18rpm. The resulting culture broth was recorded as the original fermentation broth.

[0089] (2) Preparation of 10-fold and 100-fold dilution gradient fermentation broth: Take 1 mL of the original fermentation broth and add it to 9 mL of sterile distilled water to obtain 10-fold dilution fermentation broth; take 1 mL of 10-fold dilution fermentation broth and add it to 9 mL of sterile distilled water to obtain 100-fold dilution fermentation broth.

[0090] (3) Preparation of assay plates: Using Fusarium equiseti CGMCC3.15353 grown to the logarithmic stage, a assay plate containing 10 8 PDA culture medium plates with a concentration of CFU / mL.

[0091] (4) Antibacterial effect test: Take 2 mL each of the original fermentation broth, 10-fold diluted fermentation broth, and 100-fold diluted fermentation broth of the prepared strain L1012, and soak a 6 mm diameter filter paper disc in each broth. Place the filter paper discs soaked in the three different concentrations of fermentation broth onto a PDA medium plate containing Fusarium equiseti CGMCC 3.15353 prepared in the previous step. Incubate the medium plate with the filter paper discs at 30℃ for 24 hours and observe the inhibition zone.

[0092] Results: The original fermentation broth, 10-fold diluted fermentation broth, and 100-fold diluted fermentation broth of strain L1012 all showed inhibitory activity against the root rot pathogen Fusarium equiseti CGMCC 3.15353. Figure 1 ).

[0093] V. Determination of the evolutionary position of the strain

[0094] Genomic DNA was extracted from strain L1012 of this invention and sequenced. The 16S rRNA gene sequence (SEQ ID No. 1) was then compared online in an internationally authoritative bacterial taxonomy database (http: / / www.ezbiocloud.net / ) (Kim OS, Cho YJ, Lee K, et al. 2012, Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol, 62:716-721).

[0095] The results showed that the 16S rRNA gene sequence of strain L1012 of this invention had the highest similarity to species of the genus *Pseudomonas* (Table 3). The highest similarity value of the 16S rRNA gene sequence between strain L1012 and species of *Pseudomonas* was 97.3%. This value is significantly lower than the threshold of 98.7% for distinguishing different bacterial species, and it is preliminarily inferred that this bacterium is a new species of *Pseudomonas*.

[0096] Table 3. Information on closely related bacteria of strain L1012

[0097] Scientific name of the strain strain number 16S rRNA gene similarity (%) Pseudomonas quercus hsmgli-8 97.3 Pseudomonas typographi CA3A 97.2 Pseudomonas hunanensis LV 97.2 Pseudomonas alloputida Kh7 97.1 Pseudomonas guariconensis LMG 27394 97.0 Pseudomonas ficuserectae JCM 2400 96.9 Pseudomonas oryzicola RD9SR1 96.8 Pseudomonas anuradhapurensis RD8MR3 96.8 Pseudomonas bananamidigenes BW11P2 96.8 Pseudomonas pharyngis BML-PP036 96.8 Pseudomonas soli F-279_208 96.8 Pseudomonas mosselii CIP 105259 96.8 Pseudomonas vlassakiae RW4S2 96.8 Pseudomonas caricapapayae ATCC 33615 96.7 Pseudomonas paraglycinae BML-PP023 96.7

[0098] To further clarify the phylogenetic position of the strains, a phylogenetic tree was constructed using the 16S rRNA gene sequences of type strains from closely related species in the genus *Pseudomonas*. Figure 2 To further clarify the phylogenetic position of the strain, the average nucleotide similarity (ANI) between the whole genome sequence of strain L1012 and the whole genome sequence of the closely related control strain was compared and calculated on EZbiocloud. The ANI value of strain L1012 and the closely related control strain Pseudomonas quercushsmgli-8 was calculated to be 80.7%, which is lower than the ANI value (95%) used to classify two genera (Yoon SH, Ha SM, Lim J, Kwon S, Chun J. A large-scale evaluation of algorithms to calculate average nucleotide identity. Antonie van Leeuwenhoek 2017; 110:1281-1286.). Based on the combined physiological, biochemical, and chemical characteristics, strain L1012 of this invention was identified as a new species of the genus Pseudomonas.

[0099] In summary, by identifying and comparing the phenotypic and genotypic characteristics of strain L1012 of this invention with existing Pseudomonas strains, it is fully demonstrated that strain L1012 of this invention represents a new species of the Pseudomonas genus. Based on its isolation from ginseng rhizosphere soil, this new species is named *Pseudomonas ginsengi*. Activity testing confirmed that the strain of this invention has inhibitory activity against *Fusarium equiseti*, the pathogen of root rot. This result suggests that strain L1012 of this invention has good potential for the biocontrol of root rot in medicinal plants (such as *Panax notoginseng*, ginseng, and *Atractylodes macrocephala*), and can also be used for soil remediation and soil improvement in *Panax notoginseng*, ginseng, and *Atractylodes macrocephala* planting areas.

[0100] Pseudomonas ginsengi L1012 was deposited at the China General Microbiological Culture Collection Center on October 14, 2024, with the accession number CGMCC No. 32197.

[0101] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein.

Claims

1. Pseudomonas ginsengi, whose strain number is L1012, and whose deposit number in the General Microbiological Center of China Microbiological Culture Collection is CGMCC No.32197.

2. The culture of Pseudomonas ginsengi according to claim 1, which is obtained by culturing the Pseudomonas ginsengi according to claim 1 in a bacterial culture medium.

3. A fermentation liquid or a dilution thereof obtained by inoculating the Pseudomonas ginsengi according to claim 1 into a fermentation medium and culturing the fermentation liquid.

4. A bacterial agent, characterized in that: The bacterial agent contains the Pseudomonas ginsengi according to claim 1, the culture according to claim 2 and / or the fermentation liquid according to claim 3 or a dilution thereof.

5. The bacterial agent according to claim 4, characterized in that: The microbial agent is a microbial agent for preventing and controlling root rot and / or a microbial agent for inhibiting Fusarium equisetum.

6. Use of the Pseudomonas ginsengi according to claim 1, the culture according to claim 2, the fermentation broth or its dilution according to claim 3, or the bacterial agent according to claim 4 or 5 in any of the following: (A1) inhibits Fusarium equisetum; (A2) preparing a product for inhibiting Fusarium equisetum; (A3) Prevention and treatment of root rot; (A4) Preparing a product for controlling root rot.

7. Use of the Pseudomonas ginsengi according to claim 1, the culture according to claim 2, the fermentation broth or its dilution according to claim 3, or the bacterial agent according to claim 4 or 5 in any of the following: (B1) Soil remediation in medicinal plant cultivation areas; (B2) preparing products for soil remediation in medicinal plant cultivation areas; (B3) Soil improvement in medicinal plant cultivation areas; (B4) Preparing products for improving the soil quality of medicinal plant cultivation areas.

8. A product for inhibiting Fusarium equiseti, wherein the active ingredient comprises or is the Pseudomonas ginsengi according to claim 1, or the culture according to claim 2, or the fermentation broth or its dilution according to claim 3, or the bacterial agent according to claim 4 or 5; or A product for preventing and treating root rot, wherein the active ingredient comprises or is the Pseudomonas ginsengi according to claim 1, the culture according to claim 2, the fermentation broth or its dilution according to claim 3, or the bacterial agent according to claim 4 or 5.

9. A method for inhibiting Fusarium equiseti, comprising the following steps: treating the Fusarium equiseti to be treated or its host with the Pseudomonas ginsengi according to claim 1, the culture according to claim 2, the fermentation liquid or its dilution according to claim 3, or the bacterial agent according to claim 4 or 5; or A method for preventing and controlling root rot, comprising the following steps: treating a plant to be prevented or its growth substrate with the Pseudomonas ginsengi according to claim 1, the culture according to claim 2, the fermentation liquid or its dilution according to claim 3, or the bacterial agent according to claim 4 or 5.

10. Use of the Pseudomonas ginsengi according to claim 1 in preparing the culture according to claim 2, the fermentation broth or its dilution according to claim 3, or the bacterial agent according to claim 4 or 5.

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