Synthetic microbial community with effect of resisting panax notoginseng root rot and application of synthetic microbial community

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a synthetic microbial community with the effect of resisting panax notoginseng root rot and application of the synthetic microbial community. The invention relates to a synthetic microbial community. The synthetic microbial community is prepared from neosphingomonas sp. BW11, sphingomonas sp. BW12, sphingomyza sp. BW15, Borsella sp. BG233, variovorax sp. BJ219, pseudomonas sp. BG5 and stenotrophomonas sp. BJ8. The synthetic microbial community can effectively inhibit growth of panax notoginseng root rot fungus hyphae, reduce the morbidity of panax notoginseng root rot and achieve effective prevention and treatment of panax notoginseng root rot; the synthetic microbial community is a natural microbial combination, does not contain chemical pesticide components, is green, safe and environment-friendly, and meets the development requirements of green agriculture.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically relating to a synthetic microbial community with resistance to root rot of Panax notoginseng and its application. Background Technology

[0002] Panax notoginseng is a traditional, rare, and precious Chinese herbal medicine. It is a perennial herbaceous plant belonging to the genus Panax in the family Araliaceae. It is widely used in clinical drugs such as Yunnan Baiyao, Compound Danshen Dripping Pills, and Pien Tze Huang, as well as in functional foods, dietary supplements, and beauty products. It is recognized as a valuable medicinal and edible resource. However, root rot of Panax notoginseng is a serious soil-borne disease caused by Fusarium oxysporum (Fusarium oxysporum). Fusarium oxysporum Root rot is one of the main pathogens causing this disease. Once it occurs, it is highly contagious, has a high incidence rate, and is difficult to control, earning it the nickname "plant cancer." The incidence rate of root rot in medicinal plants is generally 10-30%, but can reach 70-80% in severe cases, sometimes even 100%, leading to complete crop failure. Current control measures for root rot include sanitation and avoiding continuous cropping to reduce pathogen residues in the planting area. These measures have a certain preventative and therapeutic effect. Chemical control methods, such as soil fumigation, have also shown good control effects. However, the use of highly toxic pesticides in chemical control processes damages the rhizosphere ecosystem of Panax notoginseng and causes problems such as excessive pesticide residues and heavy metals in Panax notoginseng, affecting the safety and efficacy of the medicinal material.

[0003] Selecting beneficial rhizosphere biocontrol bacteria for disease-resistant varieties enhances the disease resistance of the rhizosphere soil, preventing root infections caused by soil-borne pathogens and providing the first line of defense for plants. Control strategies based on plant rhizosphere microorganisms have become a research hotspot in the field of soil-borne disease control due to their environmental friendliness and sustainability. However, existing research largely focuses on the development and application of single-functional strains. Although some progress has been made, actual field trials have shown that single-strain biocontrol agents often exhibit unstable efficacy and short duration of action due to limited colonization ability in the host rhizosphere, poor environmental adaptability, and disadvantage in competing for complex ecological niches, making it difficult to meet actual production needs. To address this limitation, multi-strain combined strategies and the application of artificially synthesized microbial communities are gradually becoming research hotspots. These strategies can improve the stability and persistence of disease control effects through functional complementarity and synergistic effects among microbial communities.

[0004] The information disclosed in this background section is intended only to enhance the understanding of the overall background of the invention and should not be construed as an admission or in any way implying that the information constitutes prior art known to those skilled in the art. Summary of the Invention

[0005] The purpose of this invention is to provide a synthetic microbial community with resistance to root rot of Panax notoginseng and its application, so as to solve the problems existing in the prior art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: The first objective of this invention is to provide a synthetic microbial community comprising *Sphingosine monocytogenes* BW11, *Sphingosine monocytogenes* BW12, *Sphingosine monocytogenes* BW15, *Boswellia* BG233, *Gnaphalium* BJ219, *Pseudomonas* BG5, and *Oligotrophozoites* BJ8; wherein, The neosphingosine bacterium BW11 was purchased from the China General Microbiological Culture Collection Center. Novosphingobium kunmingense Accession number: CGMCC NO. 1.12274; The *Sphingomonas* BW12 strain was *Sphingomonas yunnanensis* purchased from the China General Microbiological Culture Collection Center. Sphingomonas yunnanensis Accession number: CGMCC NO. 1.15275; The sphingosine box bacteria BW15 was a ginseng-derived sphingosine box bacteria purchased from the China General Microbiological Culture Collection Center. Sphingopyxis ginsengisoli Accession number: CGMCC NO. 1.9098; The *Boswellia* BG233 was *Boswellia sulfurosae* purchased from the China General Microbiological Culture Collection Center. Bosea thiooxidans The accession number is CGMCC NO. 1.5383; The glutathione BJ219 is *Polyborophione*. Variovorax boronicumulans BRS2-19 was deposited at the China General Microbiological Culture Collection Center on May 28, 2025, with accession number CGMCC NO.34701; The Pseudomonas BG5 is a type of Pseudomonas marginalis. Pseudomonas marginalis BRS125 was deposited on May 28, 2025 at the China General Microbiological Culture Collection Center (CGMCC), accession number: CGMCC NO.34702; The oligotrophomonas BJ8 is a strain Stenotrophomonas humi BRS348 was deposited on May 28, 2025, at the China General Microbiological Culture Collection Center (CGMCC), accession number: CGMCC NO. 34703.

[0007] Furthermore, the viable counts of the aforementioned *Sphingosine monocytogenes* BW11, *Sphingosine monocytogenes* BW12, *Sphingosine monocytogenes* BW15, *Boswellia* BG233, *Burkholderia* BJ219, *Pseudomonas* BG5, and *Oligotrophomonas* BJ8 were all 1.0 × 10⁻⁶. 8 cfu / mL.

[0008] A second objective of this invention is to provide the application of the synthetic microbial community in the resistance to root rot of Panax notoginseng.

[0009] Furthermore, the pathogen causing root rot in Panax notoginseng includes Fusarium oxysporum.

[0010] Furthermore, in combating root rot of Panax notoginseng, the synthetic microbial community solution is applied to the substrate around the roots of Panax notoginseng at a rate of 5 mL per plant, and then applied again after an interval of 5 days.

[0011] A third objective of this invention is to provide the application of the synthetic microbial community in the prevention and control of plant diseases caused by Fusarium oxysporum infection.

[0012] Furthermore, the plant disease mentioned includes root rot of Panax notoginseng.

[0013] Compared with the prior art, the present invention has the following beneficial effects: The synthetic microbial community of the present invention can effectively inhibit the mycelial growth of Panax notoginseng root rot fungus, reduce the incidence of Panax notoginseng root rot, and achieve effective prevention and control of Panax notoginseng root rot; moreover, the synthetic microbial community of the present invention is a natural microbial combination, does not contain chemical pesticide components, is green and safe, environmentally friendly, and meets the requirements of green agriculture development.

[0014] Preservation Information Boron glutathione Variovorax boronicumulans BRS2-19 was deposited on May 28, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO. 34701. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, 100101, China. Marginal Pseudomonas Pseudomonas marginalis BRS125 was deposited on May 28, 2025 at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO. 34702. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, 100101, China. strain Stenotrophomonas humi BRS348 was deposited on May 28, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO. 34703. The depository address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, 100101, China. Attached Figure Description

[0015] Figure 1 This is the result of the potted plant experiment of the present invention. Detailed Implementation

[0016] The technical solution of this invention patent will be clearly and completely described below. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without inventive effort are within the scope of protection of this invention.

[0017] 1. Materials and Methods 1.1 Test strains 1.1.1 Purchasing strains The strains *Sphingospora* BW11, *Sphingomonas* BW12, *Sphingospora Box* BW15, and *Boswellia* BG233 in this application were purchased from the China General Microbiological Culture Collection Center. Their relevant information is as follows: Neosphingosine mononitrate BW11 was purchased from the China General Microbiological Culture Collection Center in Kunming. Novosphingobium kunmingense Accession number: CGMCC NO. 1.12274; Sphingosomalidone BW12 is *Sphingosomalidone yunnanensis* purchased from the China General Microbiological Culture Collection Center. Sphingomonas yunnanensis Accession number: CGMCC NO. 1.15275; Sphingosine Box BW15 is a type of ginseng sphingosine box purchased from the China General Microbiological Culture Collection Center. Sphingopyxis ginsengisoli Accession number: CGMCC NO. 1.9098; Boswellia BG233 is a sulfur-oxidizing Boswellia sulphurea purchased from the China General Microbiological Culture Collection Center. Bosea thiooxidans Collection number: CGMCC NO. 1.5383.

[0018] 1.1.2 Isolation of bacterial strains This application isolated *Pseudomonas hummi* BJ219, *Pseudomonas* BG5, and *Oligotrophozoa* BJ8 from the roots of *Panax notoginseng*. Molecular identification methods were used, employing universal primers (Table 1) for molecular amplification and sequencing. BLAST comparisons with data from the BNCI website were then performed to determine their phylogenetic relationships. A phylogenetic tree was constructed using MEGA software to identify *Pseudomonas hummi* BJ219, *Pseudomonas* BG5, and *Oligotrophozoa* BJ8.

[0019] Table 1 Primers and Sequences

[0020] The identification result is: BJ219 is *Polyborophanthus*. Variovorax boronicumulans ; Pseudomonas BG5 is a marginal Pseudomonas. Pseudomonas marginalis ; Oligotrophomonas BJ8 is a strainStenotrophomonas humi .

[0021] 1.2 Test pathogens Fusarium oxysporum isolated from roots of Panax notoginseng diseased root rot samples ( Fusarium oxysporum ).

[0022] 1.3 Test Culture Medium LB liquid medium: 10.0 g tryptone, 5.0 g yeast extract, 5.0 g sodium chloride, 1000 mL sterile water; PDA medium: 250g potato, 20g glucose, 10g agar powder, 1000mL sterile water.

[0023] 1.3 Construction of synthetic microbial communities The tested strains *Sphingosine monocytogenes* BW11, *Sphingosine monocytogenes* BW12, *Sphingosine box bacteria* BW15, *Boswellia* BG233, *Gnaphalium affine* BJ219, *Pseudomonas* BG5, and *Oligotrophomonas* BJ8 were inoculated into LB liquid medium and cultured at 28°C and 200 rpm for 12 h. The viable cell count was then adjusted to 1.0 × 10⁻⁶. 8 cfu / mL; mix the bacterial solutions of 7 strains in equal volume ratio to obtain the synthetic microbial community SynCom.

[0024] 1.4 Flat Plate Standoff Test The inhibitory activity of bacterial suspensions from seven bacterial strains and the SynCom microbial community against the mycelial growth of the tested pathogens was determined using the plate confrontation culture method. Specifically, the tested pathogens were inoculated onto PDA medium and activated at 30°C. Then, 8 mm diameter mycelial discs were punched along the edge of the colony using a puncher. These discs were transferred to the center of a 9 cm diameter PDA plate, and 5 μL of bacterial suspension was inoculated at four points 2.5 cm from the center of the plate. Each treatment was repeated four times. A plate inoculated only with mycelial discs served as a blank control. All plates were incubated at 30°C. When the colony diameter in the blank control reached two-thirds of the plate diameter, the colony diameter was measured, and the inhibition rate was calculated. The results are shown in Table 2.

[0025]

[0026] Table 2 Results of the plate standoff test

[0027] Note: Different lowercase letters in the table represent significant differences between treatments. P <0.05, ANOVA, Duncan's method) Table 2 shows that the inhibition rates of single-strain bacterial solutions against the mycelial growth of the tested pathogen *Fusarium oxysporum* varied considerably, and were significantly lower than those of the synthetic microbial community SynCom bacterial solution. This indicates that the synthetic microbial community SynCom of the present invention can improve the inhibitory effect on *Fusarium oxysporum*, the causal agent of root rot in Panax notoginseng.

[0028] 1.5 Pot Experiment Each pot contains 4 kg of sterilized soil, and 5-6 three-year-old Panax notoginseng seedlings are transplanted into each pot. After the transplanted Panax notoginseng has recovered from transplanting, SynCom bacterial culture solution is inoculated into the soil around the Panax notoginseng plants, 5 mL per plant. The treatment is repeated once after 5 days with the same inoculation amount. A control group treated with LB liquid medium is used. Each treatment is replicated 5 times. Five days after the second SynCom inoculation, the phloem of the Panax notoginseng root is punctured with a 20 mL syringe needle, and 1×10⁻⁶ cells are injected. 7 cfu / mL F.oxysporum Inoculate the rhizosphere of Panax notoginseng with a conidial suspension at a rate of 5 mL per plant. Samples were taken 50 days after inoculation to observe disease occurrence and the proportion of diseased plants was recorded. Results are shown below. Figure 1 .

[0029] Depend on Figure 1 It can be seen that, compared with the control group inoculated only with pathogenic bacteria, inoculation with the SynCom microbial community solution can effectively reduce the incidence of root rot in Panax notoginseng. This indicates that the SynCom microbial community of the present invention effectively reduces the incidence of root rot in Panax notoginseng, achieving effective prevention and control of root rot in Panax notoginseng.

[0030] In summary, the synthetic microbial community of the present invention can effectively inhibit the mycelial growth of Panax notoginseng root rot fungus, reduce the incidence of Panax notoginseng root rot, and achieve effective prevention and control of Panax notoginseng root rot; moreover, the synthetic microbial community of the present invention is a natural microbial combination, does not contain chemical pesticide components, is green and safe, environmentally friendly, and meets the requirements of green agriculture development.

[0031] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A synthetic microbial community, characterized in that, The synthetic microbial community consists of *Neosphingosine monocytogenes* BW11, *Sphingosine monocytogenes* BW12, *Sphingosine box bacteria* BW15, *Boswellia* BG233, *Gnaphalium* BJ219, *Pseudomonas* BG5, and *Oligotrophozoites* BJ8; among which... The neosphingosine bacterium BW11 was purchased from the China General Microbiological Culture Collection Center. Novosphingobium kunmingense Accession number: CGMCC NO. 1.12274; The *Sphingomonas* BW12 strain was *Sphingomonas yunnanensis* purchased from the China General Microbiological Culture Collection Center. Sphingomonas yunnanensis Accession number: CGMCC NO. 1.15275; The sphingosine box bacteria BW15 was a ginseng-derived sphingosine box bacteria purchased from the China General Microbiological Culture Collection Center. Sphingopyxis ginsengisoli Accession number: CGMCC NO. 1.9098; The *Boswellia* BG233 was *Boswellia sulfurosae* purchased from the China General Microbiological Culture Collection Center. Bosea thiooxidans Accession number: CGMCC NO. 1.5383; The glutathione BJ219 is *Polyborophione*. Variovorax boronicumulans BRS2-19 was deposited at the China General Microbiological Culture Collection Center on May 28, 2025, with accession number CGMCC NO.34701; The Pseudomonas BG5 is a type of Pseudomonas marginalis. Pseudomonas marginalis BRS125 was deposited on May 28, 2025 at the China General Microbiological Culture Collection Center (CGMCC), accession number: CGMCC NO.34702; The oligotrophomonas BJ8 is a strain Stenotrophomonas humi BRS348 was deposited on May 28, 2025, at the China General Microbiological Culture Collection Center (CGMCC), accession number: CGMCC NO. 34703.

2. The synthetic microbial community according to claim 1, characterized in that, The viable counts of the following bacteria were all 1.0 × 10⁸: *Sphingosine monocytogenes* BW11, *Sphingosine monocytogenes* BW12, *Sphingosine cascade* BW15, *Boswellia* BG233, *Gnaphalium affine* BJ219, *Pseudomonas* BG5, and *Oligotrophomonas* BJ8. 8 cfu / mL.

3. The application of the synthetic microbial community described in claim 1 in the resistance to root rot of Panax notoginseng.

4. The application according to claim 3, characterized in that, The pathogen causing root rot in Panax notoginseng includes Fusarium oxysporum.

5. The application according to claim 3, characterized in that, Specifically, the synthetic microbial community solution is applied to the substrate around the roots of Panax notoginseng at a rate of 5 mL per plant, and then applied again after an interval of 5 days.

6. The application of the synthetic microbial community according to claim 1 in the prevention and control of plant diseases caused by Fusarium oxysporum infection.

7. The application according to claim 6, characterized in that, The plant diseases mentioned include root rot of Panax notoginseng.

Images