Pseudomonas R10 and its use

Abstract

This invention belongs to the field of agricultural biotechnology, specifically relating to a strain of Pseudomonas sp. R10 and its applications. The Pseudomonas sp. R10 provided by this invention is deposited at the China Center for Type Culture Collection (CCTCC), accession number CCTCC M 2024850. This strain exhibits significant control effects against clubroot disease, particularly in rapeseed. Verification through examples shows that Pseudomonas sp. R10 inhibits the germination rate of dormant spores of Pseudomonas by 35.14%. In pot experiments, it reduced the clubroot disease severity index by 53.06%. In field trials, treatment of rapeseed with R10 solution reduced the clubroot disease severity index by 39.45%, and significantly increased plant height, rootstock diameter, fresh weight, number of effective branches, and number of effective siliques in the main inflorescence compared to the control plot. R10 solution treatment also increased rapeseed yield by 59.86% and thousand-grain weight by 7.25%.

Description

Technical Field

[0001] This invention belongs to the field of agricultural biotechnology, specifically relating to a strain of Pseudomonas R10 and its applications. Background Technology

[0002] Clubroot disease, a soil-borne disease caused by the protozoan *Platycorrhizal* fungus, affects cruciferous vegetables. Infected crops develop abnormally swollen roots, leading to reduced yield and quality, causing significant economic losses. Controlling clubroot is extremely difficult. Chemical control is currently the most common method, primarily through soil disinfection and the use of chemical agents. However, the extensive use of chemical agents has also caused various problems, including environmental impacts, food safety concerns, pesticide damage, and pesticide residues. Biological control, on the other hand, is a scientific method that uses beneficial organisms to control harmful organisms. Due to its advantages such as being pollution-free, less prone to developing resistance, and safer, it has a promising future and is of great significance to the further development of green agriculture in my country. However, there are currently very few reports on the exploration of microbial resources and the development of inoculants for clubroot control. Summary of the Invention

[0003] The purpose of this invention is to provide a strain of Pseudomonas R10 and its application to achieve green control of clubroot disease and enrich the microbial resources for the prevention and control of clubroot disease.

[0004] This invention provides a strain of Pseudomonas sp. R10, which is deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC M 2024850.

[0005] The present invention also provides a microbial agent comprising Pseudomonas R10 as described in the above technical solution.

[0006] Preferably, the OD of the bacterial agent 600 The value ranges from 0.3 to 1.5.

[0007] The present invention also provides the application of the above-described Pseudomonas R10 or the above-described inoculum in the prevention and control of plant clubroot disease and / or the inhibition of clubroot bacteria.

[0008] Preferably, the plant includes rapeseed.

[0009] This invention also provides a method for preventing and controlling clubroot disease in rapeseed, comprising the following steps:

[0010] Rapeseeds were first treated with the microbial agent described in the above technical solution to obtain first-treated rapeseeds.

[0011] The rapeseed seeds treated with the first treatment were sown, and the rapeseed seeds were then treated with the microbial agent described in the above technical solution.

[0012] Preferably, the first treatment method includes soaking; the soaking time is 4 to 6 hours.

[0013] Preferably, the second treatment includes root irrigation;

[0014] The root irrigation is performed 3 times, with an interval of 20 to 30 days between two consecutive root irrigations.

[0015] Preferably, the first root irrigation is performed on the first day after sowing.

[0016] Preferably, the dosage of the microbial agent is 2.5–3 L / 10m² for each root drenching. 2 .

[0017] Beneficial effects:

[0018] This invention provides a strain of *Pseudomonas* R10, deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC M 2024850. This strain exhibits significant control effects against clubroot disease, particularly in rapeseed. Verification through examples shows that *Pseudomonas* R10 inhibits the germination of dormant spores of *P. clubroot* by 35.14%. In pot experiments, it reduced the clubroot disease disease index by 53.06%. In field trials, 63 days after sowing, the average disease index of the three control plots was 78.03, while that treated with strain R10 was 47.25, representing a 39.45% reduction in the disease index. 100 days after sowing, agronomic traits of rapeseed seedlings were statistically analyzed. Rapeseed treated with R10 showed significantly higher plant height, rootstock diameter, and fresh weight compared to the control plots. 207 days after field sowing, rapeseed was tested at the pod-setting stage. Treatment with R10 bacterial solution significantly increased plant height, number of effective branches, and number of effective pods in the main inflorescence compared to the control plot. 224 days after field sowing, actual rapeseed yield was measured. R10 bacterial solution treatment increased yield by 59.86% and thousand-grain weight by 7.25%.

[0019] Biological Preservation Information

[0020] Pseudomonas R10, biologically classified as Pseudomonas sp., was deposited on April 30, 2024, at the China Center for Type Culture Collection, No. 299 Bayi Road, Wuchang District, Wuhan, Hubei Province, China, with accession number CCTCC M 2024850. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0022] Figure 1The phylogenetic tree and colony morphology of strain R10;

[0023] Figure 2 Statistical results of the germination rate of dormant spores of *Pseudomonas* R10 after 3 days of treatment with fermentation broth;

[0024] Figure 3 The root swelling phenotypes of rapeseed in the control group (Pb) and treatment group (Pb+R10) 30 days after inoculation with clubroot fungus in Example 3;

[0025] Figure 4 The disease index of rapeseed in the control group (Pb) and treatment group (Pb+R10) after inoculation with clubroot fungus for 30 days in Example 3 was statistically analyzed, n=32;

[0026] Figure 5 The root diameter of rapeseed in the control group (Pb) and treatment group (Pb+R10) after 30 days of inoculation with clubroot fungus in Example 3, n=32;

[0027] Figure 6 The root weight of rapeseed in the control group (Pb) and treatment group (Pb+R10) 30 days after inoculation with clubroot fungus in Example 3, n=32;

[0028] Figure 7 The results of qRT-PCR detection of the content of Pb+R10 root phytohedrosis bacteria in rapeseed roots in Example 3;

[0029] Figure 8 The phenotypes of clubroot disease in rapeseed seedlings were observed 63 days after sowing in the control group (Pb) and treatment group (Pb+R10) in Example 4.

[0030] Figure 9 The disease index of clubroot disease in rapeseed seedlings was measured 63 days after sowing in the control group (Pb) and treatment group (Pb+R10) in Example 4.

[0031] Figure 10 The plant height of rapeseed in the control group (Pb) and treatment group (Pb+R10) 100 days after sowing in Example 4;

[0032] Figure 11 The rootstock diameter of the control group (Pb) and the treatment group (Pb+R10) rapeseed 100 days after sowing in Example 4 was measured.

[0033] Figure 12 The fresh weight of rapeseed in the control group (Pb) and treatment group (Pb+R10) 100 days after sowing in Example 4;

[0034] Figure 13 The plant height of rapeseed in the control group (Pb) and treatment group (Pb+R10) 207 days after sowing in Example 4.

[0035] Figure 14 The effective branching height of rapeseed 207 days after sowing in the control group (Pb) and treatment group (Pb+R10) in Example 4;

[0036] Figure 15 The effective number of rapeseed branches in the control group (Pb) and treatment group (Pb+R10) 207 days after sowing in Example 4;

[0037] Figure 16 The number of effective siliques in the main inflorescence of rapeseed in the control group (Pb) and treatment group (Pb+R10) 207 days after sowing in Example 4;

[0038] Figure 17 The yields of rapeseed in the control group (Pb) and treatment group (Pb+R10) 224 days after sowing in Example 4;

[0039] Figure 18 The weight of 1,000 seeds in the control group (Pb) and treatment group (Pb+R10) of rapeseed 224 days after sowing in Example 4 is shown. Detailed Implementation

[0040] This invention provides a strain of Pseudomonas sp. R10, which is deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC M 2024850.

[0041] The present invention also provides a microbial agent comprising Pseudomonas R10 as described in the above technical solution.

[0042] In this invention, the OD of the bacterial agent 600 The value is preferably 0.3 to 1.5, more preferably 0.3, 0.4 or 1.5, and even more preferably 1.5; the preparation method of the bacterial agent preferably includes: culturing Pseudomonas R10 in LB liquid medium; the culturing time is preferably 3 days; the culturing temperature is preferably 28°C.

[0043] The present invention also provides the application of the above-described Pseudomonas R10 or the above-described inoculum in the prevention and control of plant clubroot disease and / or the inhibition of clubroot bacteria.

[0044] In this invention, the plant preferably includes rapeseed; the inhibition of clubroot fungus preferably includes the inhibition of clubroot fungal spore germination.

[0045] This invention also provides a method for preventing and controlling clubroot disease in rapeseed, comprising the following steps:

[0046] Rapeseeds were first treated with the microbial agent described in the above technical solution to obtain first-treated rapeseeds.

[0047] The rapeseed seeds treated in the first treatment were sown, and then subjected to a second treatment.

[0048] This invention utilizes the microbial agent described in the above-mentioned technical solution to perform a first treatment on rapeseed seeds, obtaining first-treated rapeseed seeds. In this invention, the first treatment preferably includes soaking; the OD of the microbial agent used for soaking... 600 The value is preferably 0.3; the soaking time is preferably 4 to 6 hours, more preferably 5 hours; the soaking temperature is preferably 28°C.

[0049] After obtaining the first-treated rapeseed seeds, the present invention sows the first-treated rapeseed seeds and uses the microbial agent described in the above technical solution to perform a second treatment on the sown rapeseed seeds.

[0050] In this invention, the second treatment preferably includes root irrigation; the OD of the fungal agent used for root irrigation is... 600 The preferred value is 1.5; the preferred number of root drenching sessions is 3; the preferred time for the first root drenching is the first day after sowing; the preferred time interval between two consecutive root drenching sessions is 20-30 days; the preferred time interval between the first and second root drenching sessions is 20 days, and the preferred time interval between the second and third root drenching sessions is 30 days; the preferred dosage of the microbial agent for each root drenching session is 2.5-3 L / 10m². 2 More preferably 2.5L / 10m 2 The method provided by this invention can effectively reduce the disease index of clubroot disease in rapeseed, significantly increase the height and fresh weight of rapeseed seedlings, and effectively increase yield and thousand-grain weight.

[0051] To further illustrate the present invention, the following detailed description of a Pseudomonas R10 strain and its applications, in conjunction with the accompanying drawings and embodiments, is provided but should not be construed as limiting the scope of protection of the present invention.

[0052] The data in this embodiment of the invention were determined to be significantly different by t-test, with the error bar being the mean ± standard deviation; "*" indicates p < 0.05, "**" indicates p < 0.01, "***" indicates p < 0.001, and "****" indicates p < 0.0001.

[0053] Example 1

[0054] A bacterial strain R10 was isolated from the roots of *Rhizopus chinensis* 62R, a plant resistant to clubroot, using the dilution plating method. The bacteria were streaked onto LB agar plates and incubated at 28°C for 12 hours. Colony morphology was then observed. Figure 1 As shown in B. From Figure 1 As can be seen from B, the colony morphology of strain R10 is: milky white, viscous, with a smooth and opaque surface.

[0055] R10 was cultured overnight in liquid LB medium with shaking (LB liquid medium formulation: 10g peptone, 5g yeast extract, 5g sodium chloride, dissolved in 1000mL distilled water by heating, sterilized at 121℃ for 20min); bacterial DNA was extracted using the alkaline lysis method: 10μL of bacterial culture was added to 17μL of alkaline lysis buffer I (25mM NaOH, 0.2mM EDTA, pH 12) solution, treated at 95℃ for 30min, then 17μL of neutralization lysis buffer II (40mM Tris-HCl, pH 7.5) was added, mixed and centrifuged to obtain the DNA of strain R10.

[0056] PCR amplification of the full-length 16S rRNA from strain R10 DNA was performed using primers 27F and 1492R, followed by amplification using 2×Rapid Taq Master Mix to obtain the amplified products. The reaction system consisted of: 2 μL template DNA, 1 μL primer 27F, 1 μL primer 1492R, 20 μL mix, 16 μL DNase-free and RNase-free water, for a total volume of 40 μL. The PCR program was as follows: 95℃ pre-denaturation for 2 min; 95℃ denaturation for 20 s, 55℃ annealing for 20 s, 72℃ extension for 30 s, 35 cycles; 72℃ extension for 5 min. The PCR products were analyzed by electrophoresis on a 1% agarose gel to detect the size of the amplified target band. The primers used for amplification are as follows:

[0057] 27F: 5'-TACGGYTACCTTGTTACGACTT-3' (SEQ ID No. 1);

[0058] 1492R: 5'-AGAGTTTGATCMTGGCTCAG-3' (SEQ ID No. 2).

[0059]

[0060] Based on the results of 16S rDNA sequencing, a phylogenetic tree was constructed using the neighbor-joining method in Mega 7 software, as follows: Figure 1 As shown in Figure A.

[0061] according to Figure 1 China A, combined Figure 1 According to the results of the study, strain R10 was identified as Pseudomonas sp. and named Pseudomonas sp.R10. It was deposited on April 30, 2024, at the China Center for Type Culture Collection, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province, with accession number CCTCC M 2024850.

[0062] Example 2

[0063] Pseudomonas sp. R10 inhibits the germination of dormant spores of Pseudomonas.

[0064] 1. Dormant spores of *Plasmodiophora stylosa* were extracted and sterilized according to the method described in the literature (Bi Kai. Multi-omics study reveals the molecular mechanism of growth, development and pathogenesis of *Plasmodiophora stylosa* [D]. Huazhong Agricultural University, 2017.).

[0065] 2. Place a layer of filter paper in a 20cm diameter petri dish. After high-temperature sterilization, disinfect the rapeseed seeds with 50% (V / V) 84 disinfectant for 10 minutes. Rinse with sterile water 6-7 times and place 100 seeds in each petri dish. Add 10mL of sterile water, seal the dish, and place it in a plant culture room for cultivation. The cultivation conditions are: temperature 22℃, humidity 70%, light-dark ratio 12h light / 12h darkness. Add sterile water as needed. After 5-10 days of cultivation, collect the culture solution and filter it through a 0.22μm bacterial filter to obtain the rapeseed root exudate solution.

[0066] 3. Preparation of bacterial fermentation broth: Add 30 mL of LB liquid medium to a 50 mL sterile conical flask. Inoculate a small amount of fresh Pseudomonas sp. R10 cells from an LB culture dish using an inoculation loop. Incubate at 28 °C with shaking for 3 days. Centrifuge, collect the supernatant, and filter through a 0.22 μm bacterial filter to obtain the R10 bacterial fermentation broth.

[0067] 4. DAPI staining to observe the germination of dormant spores of *Plasmodiophora*

[0068] Dormant spores of *Plasmodiophora* were divided into a treatment group and a control group, with six 1 mL centrifuge tubes in each group. 500 μL of a 3 × 10⁻⁶ concentration was added to each tube. 7 Centrifuge clean, dormant *Plasmodiophora* spores / mL at 5000 rpm for 5 min, discard the supernatant, and then proceed as follows:

[0069] Treatment group: Add 500 μL of R10 bacterial fermentation broth and vortex mix.

[0070] Control group: Add 500 μL of root exudate and vortex mix.

[0071] Then, the mixed systems were placed in a rapeseed culture room and cultured in the dark for 3 days. Microscopic observation: 20 μL of each tube was mixed with 2 μL of DAPI dye, and slides were prepared. The slides were observed and photographed using a fully motorized upright fluorescence microscope (BX63). Six images were collected from each group. After spore counting, the inhibition rate of the strain on the germination of dormant spores of *Plasmodiophora stylosa* was calculated using the following formula. The results are as follows: Figure 2 As shown.

[0072] Spore germination inhibition rate (100%) = (germination rate of dormant spores in control group - germination rate of dormant spores in treatment group) × 100% / germination rate of dormant spores in control group.

[0073] Depend on Figure 2 It can be seen that the germination rate of the control group was 80.86%, while the germination rate of the treatment group was 52.45%, meaning that R10 inhibited the germination of dormant spores of *Plasmodiophora* by 35.14%.

[0074] Example 3

[0075] Potted experiment with Pseudomonas sp.R10

[0076] After centrifuging the R10 bacterial fermentation broth prepared in step 3 of Example 2, the bacterial cells were collected, and sterile water was added to adjust its OD. 600 Values ​​were prepared to obtain OD values. 600 =0.4 R10 bacterial culture and OD 600 Prepare R10 bacterial culture with a concentration of 1.5 for later use.

[0077] Rapeseed seeds were disinfected with 50% (v / v) 84 disinfectant for 15 minutes and rinsed 6-7 times with sterile water. They were then divided into a control group (Pb) and a treatment group (Pb+R10), with 32 seeds in each group, and treated as follows:

[0078] Treatment group (Pb+R10): Rapeseed seeds were subjected to OD... 600 After soaking in R10 bacterial solution at 0.4 for 1 hour, the rapeseed was sown. Nine days after sowing, each rapeseed plant was treated with 5 ml of R10 bacterial solution (OD). 600 =1.5) Root irrigation; On the 10th day after sowing, inoculate each rapeseed plant with 1 mL of 1×10⁻⁶ solution. 7 A solution of dormant spores of *Plasmodiophora* at spores / mL was used, followed by R10 bacterial suspension (OD) every week. 600=1.5) Drench the roots once, with each rapeseed plant receiving 5 mL of water each time, for a total of 3 weeks.

[0079] Control group (Pb): Rapeseed seeds were soaked in water for 1 hour before sowing. Nine days after sowing, each rapeseed plant was irrigated with 5 mL of water. On the 10th day after sowing, each rapeseed plant was inoculated with 1 mL of a 1×10⁻⁶ solution. 7 The plant was treated with a solution of dormant spores of *Plasmodiophora stearothermiae* at spores / mL, followed by root irrigation with water once a week, with 5mL of solution applied to each rapeseed plant each time, for a total of 3 weeks.

[0080] (1) Thirty days after rapeseed was inoculated with clubroot fungus, the disease incidence was statistically analyzed, and the disease index and control effect were calculated according to the following formula. The results are as follows: Figures 3-4 As shown.

[0081] Disease Index (DI) = (1n1 + 3n3 + 5n5) × 100 / 5N t

[0082] Prevention and control effect (%) = (disease index of control - disease index of treatment) × 100% / disease index of control;

[0083] Where n1, n3, and n5 are the number of rapeseed plants with disease severity levels of 1, 3, and 5, respectively, and N t This refers to the total number of rapeseed plants. The grading standard for clubroot disease severity used in this study is as follows: Grade 0: No tumors on the roots; Grade 1: Small tumors on the lateral roots; Grade 3: Enlarged taproot with a diameter less than twice that of the stem base; Grade 5: Enlarged taproot with a diameter 2 to 3 times that of the stem base or rotten.

[0084] Depend on Figure 3 It can be seen that the root disease incidence in the treatment group was milder than that in the control group. Figure 4 It can be seen that the disease index of the control group was 62, while that of the treatment group was 29.1, and the prevention and control effect was 53.06%; the disease index of rapeseed in the treatment group was also significantly lower than that of the control group.

[0085] (2) Root samples were taken from both the treatment group (Pb+R10) and the control group (Pb) of rapeseed. Root diameter and root weight were recorded. The results are as follows: Figure 5 and Figure 6 As shown.

[0086] Depend on Figure 5 and Figure 6 It can be seen that the root diameter and root weight of the rapeseed in the treatment group were significantly different from those in the control group.

[0087] (3) After the roots of the rapeseed in the treatment group (Pb+R10) were ground with liquid nitrogen, DNA was extracted using the CTAB method, and the content of clubroot bacteria was detected by qRT-PCR. The results are as follows: Figure 7As shown.

[0088] Depend on Figure 7 It can be seen that the content of clubroot bacteria in the roots of rapeseed in the treatment group was significantly lower than that in the control group, showing a better biocontrol effect.

[0089] Example 4

[0090] Pseudomonas sp.R10 field trial

[0091] 1. Field trial sowing and bacterial solution treatment

[0092] On September 29, 2023, 12 plots were set up in fields with severe clubroot disease in Zhijiang City, Hubei Province. Each plot was 5m long and 2m wide, with 10 rows of rapeseed planted parallel to the length, with about 20 plants per row. The plots were divided into a control group (Pb) and a bacterial solution treatment group (Pb+R10), with 6 plots in each group. Three plots in each group were used for disease index statistics during the seedling and flowering stages, and three plots were used for yield statistics.

[0093] The concentration of 3×10 was prepared according to the method described in Example 3. 9 cfu / mL (OD) 600 =1.5) of R10 bacterial culture.

[0094] Bacterial suspension treatment group (Pb+R10): Concentration used was 3×10⁻⁶. 9 cfu / mL (OD) 600 Rapeseed seeds of variety Shuang 13 were soaked in R10 bacterial solution (1.5%) and incubated at 28℃ for 5 hours, then dried for later use. Sowing began the day after seed treatment. On the day of sowing, the seeds were irrigated once with bacterial solution, 2.5L of a 3×10⁻⁶ concentration per plot. 9 cfu / mL (OD) 600 =1.5) bacterial solution, irrigate each plot once on the 20th and 50th day after sowing, with 2.5L of bacterial solution per plot each time at a concentration of 3×10 9 cfu / mL (OD) 600 =1.5) bacterial solution.

[0095] Control group (Pb): Rapeseed seeds of variety Zhongshuang 13 were soaked in sterile water, cultured at 28℃ for 5 hours, and then dried for later use. Sowing began the day after seed treatment. On the day of sowing, each plot was irrigated with tap water once, 2.5L of which was used for each plot. On the 20th and 50th days after sowing, each plot was irrigated with tap water once more, 2.5L of which was used for each plot.

[0096] 2. Statistics on rapeseed disease in field trials

[0097] Sixty-three days after sowing, 50 rapeseed plants were randomly selected from each group in a plot used to collect disease index data. The incidence and disease index of clubroot disease in rapeseed seedlings were statistically analyzed using the following methods. The results are as follows: Figure 8 and Figure 9 .

[0098] Disease Index (DI) = (1n1 + 2n2 + 3n3 + 4n4) × 100 / 4N t

[0099] Where n1-n4 represent the number of rapeseed plants with disease severity levels 1 to 4, and N t This refers to the total number of rapeseed plants. Based on the severity of the disease, the condition is divided into five levels: 0-4. Level 0: No tumors on the roots, normal root development; Level 1: No disease on the main root, small tumors on some lateral roots and fibrous roots; Level 2: Mild disease on the main root, slightly swollen, obvious tumors on some lateral roots and fibrous roots; Level 3: Severe disease on the main root, abnormally swollen and cracked, obvious tumors on most lateral roots and fibrous roots, but the plant grows normally; Level 4: Almost no fibrous roots, the main root is rotten or dead, the plant is wilted or dead.

[0100] Depend on Figure 8 and Figure 9 It can be seen that the disease index of rapeseed in the control group (Pb) was 78.03, while the disease index of the bacterial suspension treatment group (Pb+R10) was 47.25. The biocontrol effect of R10 strain on rapeseed clubroot disease can reach 39.45%.

[0101] 3. Statistical analysis of agronomic traits of rapeseed in field trials

[0102] One hundred days after sowing, the agronomic traits of rapeseed seedlings were statistically analyzed. Eight rapeseed plants were randomly selected from each plot used for seedling disease index calculation in each group, totaling 24 plants from three plots in each group. Plant height, rootstock diameter, and fresh weight were then statistically analyzed. The results are as follows: Figures 10-12 As shown.

[0103] Depend on Figures 10-12 It can be seen that, compared with the control group (Pb), the rapeseed treated with the bacterial solution (Pb+R10) showed a significant increase in plant height, root and stem diameter and fresh weight.

[0104] 4. Field Trial Rapeseed Yield Statistics

[0105] (1) 207 days after sowing, during the rapeseed pod-bearing stage, 8 rapeseed plants were randomly selected from each plot used to count the disease index during the seedling and flowering stages in each group. A total of 24 rapeseed plants were selected from 3 plots in each group. The plant height, effective branch height, number of effective branches, and number of effective pods in the main inflorescence were counted. Figures 13-16 As shown.

[0106] Among them, the effective branching height is the length from the uppermost effective silique at the top of the main inflorescence to the effective silique at the base of the main inflorescence, and the effective silique number of the main inflorescence refers to the number of siliques with more than one normal seed on the main inflorescence.

[0107] Depend on Figure 13 , 15 As shown in Figure 16, compared with the control group (Pb), the rapeseed treated with the bacterial solution (Pb+R10) showed significantly increased plant height, number of effective branches, and number of effective siliques in the main inflorescence after treatment with R10 bacterial solution. Figure 14 It can be seen that there was no significant difference in the effective branching height between the bacterial culture treatment group (Pb+R10) and the control group (Pb).

[0108] (2) 224 days after sowing, the actual yield of rapeseed was measured, and data were collected from each plot (10m²) used for yield statistics. 2 The rapeseed was dried, and the total yield and thousand-grain weight of each plot were measured. The results are as follows: Figure 17 and Figure 18 As shown.

[0109] Depend on Figure 17 and Figure 18 It can be seen that the yield of the control group (Pb) was 93 g / m³. 2 The yield of the bacterial culture treatment group (Pb+R10) was 148.67 g / m³. 2 Compared to the control group (Pb), the concentration of Pb increased by 59.86%; the thousand-grain weight of the control group (Pb) was 4.69 g, and the thousand-grain weight of the bacterial culture treatment group (Pb+R10) was 5.03 g, which was 7.25% higher than that of the control group (Pb).

[0110] As can be seen from the above embodiments, the Pseudomonas R10 provided by the present invention has the potential to prevent clubroot disease in rapeseed.

[0111] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A strain of Pseudomonas sp. R10, deposited at the China Center for Type Culture Collection, accession number CCTCC M 2024850.

2. A microbial agent, characterized in that, The bacterial agent includes Pseudomonas R10 as described in claim 1.

3. The microbial agent according to claim 2, characterized in that, The OD of the bacterial agent 600 The value is 0.3 to 1.

5.

4. The application of the Pseudomonas R10 of claim 1 or the inoculum of claim 2 or 3 in the prevention and control of clubroot disease and / or inhibition of clubroot bacteria; wherein the plant is rapeseed.

5. A method for controlling clubroot disease in rapeseed, characterized in that, Includes the following steps: Rapeseeds are first treated with the microbial agent described in claim 2 or 3 to obtain first-treated rapeseeds. The rapeseed seeds treated with the first treatment were sown, and the sown rapeseed seeds were then subjected to a second treatment using the microbial agent described in claim 2 or 3.

6. The method according to claim 5, characterized in that, The first treatment method includes soaking; the soaking time is 4 to 6 hours.

7. The method according to claim 5, characterized in that, The second treatment method includes root irrigation; The root irrigation is performed 3 times, with an interval of 20-30 days between two consecutive root irrigations.

8. The method according to claim 7, characterized in that, The first root irrigation should be done on the first day after sowing.

9. The method according to claim 8, characterized in that, Each time the roots are irrigated, the dosage of the microbial agent is 2.5~3L / 10m². 2 .

Images