Pseudomonas bacterial composition, composite bacterial agent and application of pseudomonas bacterial composition and composite bacterial agent

Abstract

The invention provides a pseudomonas bacterium composition, a complex microbial inoculant and application of the pseudomonas bacterium composition and the complex microbial inoculant, and belongs to the technical field of microorganisms. The invention provides a Pseudomonas bacteria combination, which comprises Pseudomonas bacteria JF1 and Pseudomonas hutmensis JF2, the Pseudomonas bacteria JF1 and the Pseudomonas hutmensis JF2 are growth-promoting bacteria screened from rhizosphere soil of rhizoma corydalis, and the Pseudomonas bacteria JF1 and the Pseudomonas bacteria JF2 have the capabilities of dissolving phosphorus and producing siderophores, can be used as growth promoting agents in a synergistic manner to promote the growth of rhizoma corydalis seedlings, and can be used for promoting the growth of rhizoma corydalis seedlings. The method has a remarkable effect of promoting accumulation of the total alkaloid content in rhizoma corydalis tubers, can be used for artificial cultivation of rhizoma corydalis, provides a theoretical basis for improving the yield and quality of rhizoma corydalis, and has important guiding significance for reasonably and effectively utilizing microorganisms to improve the quality and yield of Qinshi medicine-rhizoma corydalis.

Description

Technical Field

[0001] The present invention belongs to the technical field of microorganisms, and in particular relates to a Pseudomonas bacterial combination, a composite bacterial agent and applications thereof. Background Art

[0002] Corydalis yanhusuo ( Corydalis yanhusuo WTWang) is a wonderful product for promoting blood circulation, removing blood stasis, promoting qi circulation, and relieving pain, and is particularly renowned for its analgesic properties. Corydalis yanhusuo (Rhizoma Corydalis) is a plant that prefers warm, humid, and cool climates and is best cultivated in humus-rich, loose, well-drained sandy loam. The size of Corydalis yanhusuo tubers is an important indicator for evaluating their quality; larger tubers indicate better quality and a higher market price. Corydalis yanhusuo tubers contain alkaloids, which have strong analgesic, sedative, and hypnotic effects. The higher the alkaloid content in the tubers, the better the quality. Rhizosphere microorganisms are the second genome of plants and play a vital role in their growth and development. How to utilize rhizosphere growth-promoting bacteria to promote the growth of Corydalis yanhusuo and increase the content of its key active ingredient, total alkaloids, is a technical issue that currently needs to be addressed. Summary of the Invention

[0003] The purpose of the present invention is to provide a Pseudomonas bacteria combination, a composite bacterial agent and their application. The Pseudomonas bacteria combination and the composite bacterial agent of the present invention can not only promote the growth of Corydalis yanhusuo, but also increase the content of total alkaloids, the key active ingredient of Corydalis yanhusuo.

[0004] The present invention provides a Pseudomonas bacteria combination, including Pseudomonas bacteria deposited in CGMCC on June 11, 2025 ( Pseudomonas arcuscaelestis ) JF1 and Pseudomonas bacteria deposited in CGMCC on June 11, 2025 ( Pseudomonas hutmensis ) JF2; the preservation number of the Pseudomonas bacterium JF1 is CGMCC NO.34846; the preservation number of the Pseudomonas bacterium JF2 is CGMCC NO.34847.

[0005] Preferably, the ratio of the effective viable counts of the Pseudomonas bacteria JF1 to the Pseudomonas bacteria JF2 is 1:1.

[0006] The present invention also provides a composite bacterial agent, comprising the Pseudomonas bacteria combination described in the above scheme.

[0007] Preferably, the composite bacterial agent comprises a bacterial suspension of Pseudomonas bacteria JF1 and a bacterial suspension of Pseudomonas bacteria JF2; the volume ratio of the bacterial suspension of Pseudomonas bacteria JF1 and the bacterial suspension of Pseudomonas bacteria JF2 is 1:1; the OD of the bacterial suspension of Pseudomonas bacteria JF1 and the bacterial suspension of Pseudomonas bacteria JF2 is 600 are 0.8 respectively.

[0008] The present invention also provides the use of the Pseudomonas bacteria combination or the composite bacterial agent described in the above scheme in promoting the growth of Corydalis yanhusuo and / or increasing the total alkaloid content in Corydalis yanhusuo tubers.

[0009] Preferably, the promoting the growth of Corydalis yanhusuo includes promoting the growth of Corydalis yanhusuo seedlings and / or promoting the growth of Corydalis yanhusuo tubers; and the plant includes Corydalis yanhusuo.

[0010] Preferably, the method of promoting the growth of Corydalis yanhusuo seedlings includes at least one of 1) and 2): 1) Promote the growth of the above-ground and / or underground parts of Corydalis yanhusuo seedlings; 2) Increase the fresh weight and / or dry weight of the entire Corydalis yanhusuo seedling; The method for promoting the growth of Corydalis tubers includes at least one of (1) and (2): (1) Increase the transverse and / or longitudinal diameter of the tuber of Corydalis yanhusuo; (2) Increase the fresh weight and / or dry weight of Corydalis tubers.

[0011] Preferably, the promotion of the growth of Corydalis yanhusuo is achieved by dissolving inorganic phosphorus and / or secreting siderophores through a combination of Pseudomonas bacteria.

[0012] The present invention also provides a method for promoting the growth of Corydalis yanhusuo and / or increasing the total alkaloid content in Corydalis yanhusuo tubers, comprising the following steps: applying the Pseudomonas bacterial combination or the composite bacterial agent described in the above scheme to Corydalis yanhusuo seedlings.

[0013] Preferably, the application method includes root irrigation.

[0014] The present invention provides a Pseudomonas bacteria combination, comprising Pseudomonas bacteria ( Pseudomonas arcuscaelestis )JF1 and ( Pseudomonas hutmensis ) JF2; the Pseudomonas bacteria JF1 has a deposit number of CGMCC NO.34846; the Pseudomonas bacteria JF2 has a deposit number of CGMCC NO.34847. In the present invention, the Pseudomonas bacteria JF1 and the Pseudomonas bacteria JF2 are growth-promoting bacteria screened from the rhizosphere soil of Corydalis yanhusuo. Pseudomonas bacteria JF1 and Pseudomonas bacteria JF2 have the ability to solubilize phosphate and produce siderophores. They can be used synergistically as growth promoters to promote the growth of Corydalis yanhusuo seedlings, especially the accumulation of total alkaloids in Corydalis yanhusuo tubers. They can be used in the artificial cultivation of Corydalis yanhusuo, provide a theoretical basis for improving the yield and quality of Corydalis yanhusuo, and have important guiding significance for the rational and effective use of microorganisms to improve the quality and yield of "Qin medicine" - Corydalis yanhusuo.

[0015] Biological Deposit Description Pseudomonas bacteria (Pseudomonas sp.) Pseudomonas arcuscaelestis ) JF1, deposited on June 11, 2025 at the China General Microbiological Culture Collection Center (CGMCC), located at No. 1, Huayuancun, Beijing, China, and assigned accession number CGMCC No. 34846.

[0016] Pseudomonas bacteria (Pseudomonas sp.) Pseudomonas hutmensis ) JF2, deposited on June 11, 2025 at the China General Microbiological Culture Collection Center (CGMCC), located at No. 1, Huayuancun, Beijing, China, and assigned accession number CGMCC No. 34847. BRIEF DESCRIPTION OF DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiments will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0018] Figure 1 Colony characteristics of JF1 and JF2 bacteria; wherein A is JF1 ( Pseudomonas arcuscaelestis ), B is JF2 ( Pseudomonas hutmensis ); Figure 2 Gram staining results of JF1 and JF2 bacteria; wherein A is JF1 ( Pseudomonas arcuscaelestis ), B is JF2 ( Pseudomonas hutmensis ); Figure 3 Phosphorus solubilizing properties of JF1 and JF2; wherein A is JF1, B is JF2; Figure 4 Iron carrier production properties of JF1 and JF2; wherein A is JF1, B is JF2; Figure 5 IAA production properties of JF1 and JF2; wherein A is JF1, B is JF2; Figure 6 Verification of growth promotion effect of JF1 and JF2; wherein A is the growth state of seedlings before irrigation; B is a comparison chart of growth states of multiple pots, the left 6 pots are the growth state of seedlings of the JF1+JF2 treatment group, and the right 6 pots are the growth state of seedlings of the control TSA treatment group; C is a comparison chart of growth states of individual pots, the left side is the growth state of seedlings of the JF1+JF2 treatment group after irrigation for 30 days, and the right side is the growth state of seedlings of the control TSA treatment group after irrigation for 30 days; Figure 7This is a comparison of the whole-plant Corydalis yanhusuo seedlings in the JF1+JF2 treatment group and the TSA control group 30 days after root irrigation treatment; A is the TSA control group and B is the JF1+JF2 treatment group; Figure 8 This is a phenotypic comparison of Corydalis yanhusuo seedlings and tubers in the JF1+JF2 bacteria treatment group and the TSA control group 30 days after root irrigation. Figure 9 This is a comparison of the total alkaloid content in the tubers of Corydalis yanhusuo in the JF1+JF2 bacteria treatment group and the TSA control group 30 days after root irrigation. DETAILED DESCRIPTION

[0019] The present invention provides a Pseudomonas bacteria combination, comprising Pseudomonas bacteria ( Pseudomonas arcuscaelestis )JF1 and ( Pseudomonas hutmensis ) JF2; the preservation number of the Pseudomonas bacteria JF1 is CGMCC NO.34846; the preservation number of the Pseudomonas bacteria JF2 is CGMCC NO.34847.

[0020] In the present invention, the Pseudomonas bacteria JF1 and Pseudomonas bacteria JF2 are screened from the rhizosphere soil of Corydalis yanhusuo experimental planting base in Shangyuanguan Town, Chenggu County, Hanzhong City, Shaanxi Province.

[0021] In the present invention, the Pseudomonas bacteria JF1 and the Pseudomonas bacteria JF2 have strong phosphate solubilization ability and siderophore production ability; the Pseudomonas bacteria combination has a significant promoting effect on the growth of the aboveground parts and tubers of Corydalis seedlings, can effectively promote the growth of Corydalis seedlings, and has a significant promoting effect on the accumulation of total alkaloid content in Corydalis tubers.

[0022] In the present invention, the nucleotide sequence of 16S rDNA of the Pseudomonas bacteria JF1 is shown as SEQ ID NO.1.

[0023] In the present invention, the nucleotide sequence of 16S rDNA of the Pseudomonas bacteria JF2 is shown as SEQ ID NO.2.

[0024] In the present invention, the Pseudomonas bacteria JF1 and JF2 colonies are bright yellow, smooth, and slightly convex in the center. Gram staining reveals them to be Gram-negative bacteria. Oil immersion microscopy (100x magnification) reveals that JF1 and JF2 strains are rod-shaped.

[0025] In the present invention, the Pseudomonas bacteria JF1 and JF2 do not fix nitrogen and do not secrete auxin.

[0026] As an implementation form, the ratio of the effective viable cell number of the Pseudomonas bacteria JF1 to the effective viable cell number of the Pseudomonas bacteria JF2 is 1:1.

[0027] The application further provides a composite microbial agent, comprising the Pseudomonas bacteria combination according to the above-mentioned scheme.

[0028] As an implementation form, the composite microbial agent comprises a bacterial suspension of the Pseudomonas bacteria JF1 and a bacterial suspension of the Pseudomonas bacteria JF2; the volume ratio of the bacterial suspension of the Pseudomonas bacteria JF1 to the bacterial suspension of the Pseudomonas bacteria JF2 is 1:1; the OD600 of the bacterial suspension of the Pseudomonas bacteria JF1 is 0.8, and the OD600 of the bacterial suspension of the Pseudomonas bacteria JF2 is 0.8. 600

[0029] As an implementation form, the preparation method of the bacterial suspension of the Pseudomonas bacteria JF1 and the bacterial suspension of the Pseudomonas bacteria JF2 comprises the following steps: The Pseudomonas bacteria JF1 and the Pseudomonas bacteria JF2 are inoculated into TSA liquid culture medium respectively, and are subjected to shock culture to obtain bacterial suspensions, and the OD600 of the bacterial suspensions is adjusted to 0.8; the shock culture is carried out at a temperature of 25℃, at a rotation speed of 200 rpm, and for 1 day. 600

[0030] The application further provides an application of the Pseudomonas bacteria combination according to the above-mentioned scheme or the composite microbial agent in promoting the growth of Corydalis and / or increasing the total alkaloid content in the tuber of Corydalis.

[0031] As an implementation form, the promotion of the growth of Corydalis comprises promotion of the growth of Corydalis seedlings and / or promotion of the growth of the tuber of Corydalis; the plant comprises Corydalis.

[0032] As an implementation form, the promotion of the growth of Corydalis seedlings comprises at least one of the following 1) and 2): 1) promotion of the growth of the aboveground part and / or the underground part of the Corydalis seedling; 2) increase of the fresh weight and / or the dry weight of the whole Corydalis seedling; The promotion of the growth of the tuber of Corydalis comprises at least one of the following 1) and 2): 1) increase of the transverse diameter and / or the longitudinal diameter of the tuber of Corydalis; 2) increase of the fresh weight and / or the dry weight of the tuber of Corydalis.

[0033] As an implementation form, the promotion of the growth of Corydalis is achieved by the Pseudomonas bacteria combination dissolving inorganic phosphorus and / or secreting iron carriers.

[0034] ​​The application further provides a method for promoting the growth of Corydalis van- houttei and / or increasing the total alkaloid content in the tubers of Corydalis vanhouttei, comprising the following steps: Applying the Pseudomonas bacterial combination or the complex microbial agent described in the above scheme to the seedlings of Corydalis vanhouttei.

[0035] As an implementation form, the application mode comprises root irrigation.

[0036] As an implementation form, the application amount of the root irrigation is 30 mL / plant in total, in terms of the bacterial suspension of Pseudomonas bacterial JF1 and the bacterial suspension of Pseudomonas bacterial JF2; the OD600 of the bacterial suspension of Pseudomonas bacterial JF1 and the bacterial suspension of Pseudomonas bacterial JF2 is 0.8 respectively. 600 0.8 respectively.

[0037] In order to further illustrate the application, the Pseudomonas bacterial combination, the complex microbial agent and the application thereof provided by the application are described in detail below in combination with the drawings and examples, but they should not be understood as limiting the protection scope of the application.

[0038] Example 1 I. Strain isolation and screening TSA solid culture medium preparation: weigh 15 g of tryptone, 5 g of soybean peptone, 5 g of sodium chloride and 15 g of agar, add water to 1 L, adjust the pH to 7.3±0.2, and sterilize at 121℃ for 15 min. The liquid medium used in the experiment is the formula of the solid culture medium without agar.

[0039] In the Corydalis vanhouttei experimental planting base of Shangyuan Town, Chenggu County, Hanzhong City, Shaanxi Province, the five-point sampling method was used, 20 plants of Corydalis vanhouttei were selected at each point, the Corydalis vanhouttei plants were dug out together with the soil, the large soil around the tubers was shaken off, about 1 mm of soil adhering to the surface of the tubers was left, then a proper amount of undamaged tubers were selected and placed in a sterilized plastic bottle, about 100 mL of PBS buffer was used to wash the soil adsorbed on the surface of the tubers, and the soil suspension was placed in a 250 mL conical flask and shaken (25℃, 120 r / min) for 1 h, and then treated with 140 kHZ ultrasonic for 40 s.

[0040] 1 mL of the soil suspension was diluted to 10 -4 times by stages, and 200 μL of the diluted 10 -4The soil suspension with a certain concentration was coated on TSA medium, and after the surface of the medium was air-dried, it was inverted and cultured in a thermostat at 25°C for 16-18 h. New single colonies were picked, and the process was repeated until no new colonies were grown. The grown bacterial colonies were transferred to corresponding 1 mL liquid medium and placed in a thermostat shaker for overnight culture (200 rpm, 25°C). After three-zone streaking and multiple purification, bacterial pure culture was obtained. The morphology and characteristics of the bacteria on the medium were used for preliminary classification and removal of duplicates. The purified bacteria were named JF1 and JF2.

[0041] II. Strain identification 1. Identification and preservation of JF1 and JF2 strains Primers 27F (5'-agagtttgatcctggctcag-3', SEQ ID NO. 3) and 1492R (5'- tacggttaccttgttacgactt-3', SEQ ID NO. 4) were used for PCR amplification of 10 replicates of JF1 and JF2 bacterial liquid cultured overnight on a shaker, and then sent for identification. The PCR reaction system (25 μL) was as follows: 1 μL of each primer, 1 μL of JF1 and JF2 bacterial liquid, respectively, 2×Taq PCR Mix 12.5 μL, and ddH2O to make up to 25 μL. The PCR reaction conditions were as follows: first denaturation at 94°C for 3 min, then 35 cycles of denaturation at 94°C for 25 s, annealing at 55°C for 25 s, and extension at 72°C for 1.5 min, and finally extension at 72°C for 5 min. 5 μL of the removed PCR product was electrophoresed on a 1% agarose gel, and the PCR product with a bright band at 1500 bp was sent for sequencing. The sequencing results were compared with the EzBioCloud database (https: / / www.ezbiocloud.net / ). 800 μL of JF1 and JF2 bacterial liquid was added with 800 μL of 50% sterile glycerol and mixed, and then preserved at -80°C. The comparison results showed that JF1 was Pseudomonas arcuscaelestispseudomonas hutmensis

[0042] 2. Morphological identification of the strains According to the colony characteristics of the purified culture of the isolated strains, such as Figure 1 , as shown in Table 1: the colonies of the JF1 and JF2 bacteria were bright yellow, smooth on the surface, and slightly convex round. After Gram staining, they were Gram-negative bacteria, and under oil lens observation (100 times), the JF1 and JF2 strains were bacilli (as shown in Table 1). Figure 2

[0043] Table 1 Colony morphology of JF1 and JF2

[0044] 3. Physiological and biochemical identification Referring to the Common Bacteria System Identification Manual, the physiological and biochemical characteristics of the JF1 and JF2 bacteria were analyzed and determined: the results of Gram staining, sugar decomposition, V-P determination, methyl red, citrate utilization, starch hydrolysis, indole, contact enzyme, oxidase, hydrogen sulfide, and gelatin liquefaction experiments are shown in Table 2.

[0045] Table 2 Physiological and biochemical characteristics of JF1 and JF2

[0046] III. Functional identification of the strains 1. Nitrogen fixation The Ashby nitrogen-free culture medium was used to analyze the nitrogen fixation of the JF1 and JF2 bacteria. Based on the characteristics of the Ashby culture medium, only the strains with nitrogen fixation can grow normally on the medium. The activated bacterial suspension (OD 600 = 0.5) of the JF1 and JF2 bacteria was added into the Ashby liquid culture medium (1:50 / v:v), and cultured at 25°C, 200 rpm in a constant temperature shaker for 1d. Then the bacterial liquid was plated and streaked on the Ashby solid culture medium for continuous subculture for 3 times. The strains could grow normally, which was considered to have nitrogen fixation. (5 replicates) The results showed that the JF1 and JF2 bacteria could not grow normally, indicating that the two strains had no nitrogen fixation.

[0047] 2. Phosphorus solubilization ​The PVK phosphorus-solubilizing medium (glucose 10 g, (NH4)2SO40.5 g, manganese sulfate 0.03 g, FeSO4·7H2O 0.03 g, sodium chloride 0.3 g, magnesium sulfate heptahydrate 0.3 g, KCl 0.3 g, Ca3(PO4)22.5 g, agar 15 g, add water to 1 L (pH = 7.4 ± 0.2), 121 ℃ high-pressure sterilization for 15 min) was used for JF1 and JF2 bacteria to analyze the phosphorus-solubilizing effect. 2 μL of activated JF1 and JF2 bacterial suspensions (OD 600 = 0.5) were respectively inoculated on the PVK phosphorus-solubilizing detection medium, and then placed in darkness at 25 ℃ for 7 d. Whether transparent phosphorus-solubilizing rings were formed around the colonies was observed to determine whether the strains had phosphorus-solubilizing effect. The ratio of the transparent ring diameter (D) to the colony diameter (d) indicated the strength of the strain. The blank LB solid medium (tryptone 10 g, yeast powder 5 g, sodium chloride 10 g, agar 15 g, add water to 1 L, 121 ℃ high-pressure sterilization for 15 min) was used as a control group, and 5 replicates were performed. The results showed that transparent rings were formed around JF1 and JF2 bacteria, and the average values of the phosphorus-solubilizing ring ratio (D / d) were 2.6 and 3.2, respectively, indicating that the two strains had strong phosphorus-solubilizing effect. As Figure 3 , shown in Table 3.

[0048] Table 3 JF1 and JF2 strain phosphorus-solubilizing diameter determination

[0049] 3. Iron carrier production effect The CAS (chromazurol S) medium (purchased from Qingdao Haibo Biotechnology Co., Ltd., product number HB9132, 10.87 g was weighed into 1000 mL of sterile water, boiled to completely dissolve, 121 ℃ high-pressure sterilization for 15 min, mixed, and then poured into a flat plate) was used for JF1 and JF2 bacteria to analyze the iron carrier production effect. 2 μL of activated JF1 and JF2 bacterial suspensions (OD 600 = 0.5) were respectively inoculated on the CAS solid medium, and then placed in darkness at 25 ℃ for 7 d. Whether yellow halos were formed around the colonies was observed to determine whether the strains had iron carrier production effect. The ratio of the halo diameter (D) to the colony diameter (d) indicated the strength of the strain. The blank LB solid medium (tryptone 10 g, yeast powder 5 g, sodium chloride 10 g, agar 15 g, add water to 1 L, 121 ℃ high-pressure sterilization for 15 min) was used as a control group, and 5 replicates were performed. The results showed that yellow halos were formed around JF1 and JF2 bacteria, and the average values of the halo ratio (D / d) were 4.04 and 4.38, respectively, indicating that JF1 and JF2 bacteria had strong iron carrier production effect. As Figure 4, as shown in Table 4.

[0050] Table 4 Determination of iron-chelate yellow halo diameter of JF1 and JF2 strains

[0051] 4. Auxin (IAA, indole-3-acetic acid) production detection Salkowski colorimetric method was used for qualitative analysis of IAA production by J-2 bacteria. The activated single colonies of JF1 and JF2 were inoculated into LB liquid medium without L-tryptophan (purchased from Shanghai Yuan Ye Biotechnology Co., Ltd., item number S20082) and LB liquid medium containing L-tryptophan (the content of L-tryptophan was 1.0 g / L), and cultured at 25°C, 200 rpm for 1 day. The LB liquid fermentation medium without L-tryptophan was used as a control. The fermentation broth was centrifuged at 8000 rpm for 10 min, and 2 mL of the supernatant was mixed with 2 mL of Salkowski colorimetric solution (purchased from Feijing Biotechnology Co., Ltd., item number PH1941) and incubated in the dark for 30 min. The color change of the solution was observed to determine whether the strain had the ability to produce IAA. The results showed that the color of the solution did not change red before and after treatment of JF1 and JF2, indicating that JF1 and JF2 did not produce auxin, so no further quantitative analysis was performed. For example Figure 5 , as shown in Table 5.

[0052] Table 5 Verification of IAA production characteristics of JF1 and JF2

[0053] Note: + represents a positive reaction; - represents a negative reaction.

[0054] Example 2 Evaluation of the growth-promoting effect of the strains by potting Corydalis fumarioides seedlings 1. Root drenching with bacterial solution The identified JF1 ( Pseudomonas arcuscaelestis ) and JF2 ( Pseudomonas hutmensis ) were inoculated into TSA liquid medium and cultured at 25°C, 200 rpm for 1 day. The obtained bacterial suspension was adjusted to OD 600 =0.8 for subsequent root drenching with bacterial solution. The control treatment was TSA liquid medium treated in the same way.

[0055] 2. Sample selection and treatment Select about 30 fumitory seeds with appropriate size and full granules, and carry out low-temperature germination treatment. After germination, fumitory pot planting is carried out (all fumitory seeds are subjected to 75% ethanol disinfection treatment before planting, and the soil used is subjected to 121°C high-pressure sterilization for 15 min, one fumitory is planted in each pot, and the same weight of sterilized soil is added), and after all germination, 15 fumitory seedlings with consistent growth and health conditions are selected from each group, and are labeled as JF1, JF2, JF1+JF2, and TSA control.

[0056] 3. Verification of growth promotion effect On the 8th day after the germination of fumitory seedlings in the pot (the growth state is shown in A of Figure 6 ), 30 mL of root irrigation liquid is used to irrigate the roots of fumitory seedlings, and the same amount of sterile TSA liquid medium is used for the same treatment as a control group (TSA control group). After the root irrigation treatment, the fumitory seedlings are placed in a sterile light incubator for natural growth, and sterile water is poured once every 3 days during the growth period, and the day and night temperatures are 22°C and 18°C respectively, the light duration is 14 h, and the dark duration is 10 h. After 30 days of root irrigation treatment, the length of the aboveground part and the underground part of the fumitory plant, the fresh weight and dry weight of the whole plant, the transverse diameter and longitudinal diameter of the tuber, the fresh weight and dry weight of the tuber, and the total alkaloid content of the fumitory tuber after treatment are measured (the growth state after treatment is shown in B and C of Figure 6 ). (The aboveground part: the height of the top of each fumitory plant to the soil surface of the flowerpot is measured by a ruler; the underground part: after the aboveground part of the fumitory plant is measured, the aboveground part is cut off with scissors, and the underground part is dug out, and the length of the underground part is measured by a ruler; dry weight determination: after washing the soil, freeze-drying is carried out, and the dry weight is measured after the end; total alkaloid content (3 repeats): the total alkaloid content determination kit is purchased from BaiShi Biological Technology Co., Ltd., and the item number is BL1762B).

[0057] 4. Result analysis Figure 7 The whole plant comparison chart of fumitory seedlings in the JF1+JF2 treatment group and the TSA control group after 30 days of root irrigation treatment is shown in . It can be seen from the figure that after the root irrigation treatment with the JF1+JF2 liquid, the overall growth state of the fumitory seedlings is better than that of the fumitory seedlings treated only with the TSA medium. Through observation, the aboveground part of the fumitory seedlings treated with the liquid is longer than that of the TSA medium treatment group; Figure 8and Table 6, Table 7 shows the JF1+JF2 experimental group and TSA control group after 30 days of root irrigation processing rhizome seedling aboveground part, underground part length, whole plant fresh weight, dry weight, tuber diameter, longitudinal diameter, fresh weight, dry weight comparison. From the chart, after using JF1+JF2 bacterial solution to root irrigation, the aboveground part, underground part length, whole plant fresh weight, dry weight, tuber diameter, longitudinal diameter, fresh weight, dry weight of rhizome seedling is better than that of rhizome treated with TSA medium, especially the growth promoting effect of aboveground part is significant, which is increased by 31.95% (p<0.05) compared with TSA control group; Figure 9 and Table 8 shows the total alkaloid content of JF1+JF2 bacteria treated group and TSA control group after 30 days of root irrigation processing rhizome tuber, the results show that the total alkaloid content of rhizome tuber treated by JF1+JF2 bacteria is significantly improved (p<0.05). Compared with TSA control group, it is increased by 163.73%. The results of Table 6, Table 7, Table 8 also further show that the fresh weight and dry weight of rhizome seedling, the diameter, longitudinal diameter, fresh weight, dry weight, total alkaloid of rhizome tuber are significantly improved after JF1+JF2 bacteria combined application. The above results show that JF1+JF2 bacterial strain combined treatment can effectively promote the growth of rhizome seedling.

[0058] Table 6 Phenotype comparison of JF1+JF2 bacteria treated group and TSA control group after 30 days of root irrigation processing rhizome seedling

[0059] Table 7 Phenotype comparison of JF1+JF2 bacteria treated group and TSA control group after 30 days of root irrigation processing rhizome tuber

[0060] Table 8 Comparison of total alkaloid content after JF1+JF2 treatment

[0061] Although the above embodiment makes a detailed description of the present application, it is only a part of the embodiment of the present application, not all the embodiments, and other embodiments can be obtained according to the present embodiment without creativity, which belongs to the protection scope of the present application.

Claims

1. A Pseudomonas bacterial combination, characterized in that Including Pseudomonas bacteria deposited in CGMCC on June 11, 2025 ( Pseudomonas arcuscaelestis ) JF1 and Pseudomonas bacteria deposited in CGMCC on June 11, 2025 ( Pseudomonas hutmensis )JF2; The deposit number of the Pseudomonas bacteria JF1 is CGMCC NO.34846; the deposit number of the Pseudomonas bacteria JF2 is CGMCC NO.34847.

2. The Pseudomonas bacterial combination according to claim 1, characterized in that The ratio of the effective viable bacteria count of the Pseudomonas bacteria JF1 to that of the Pseudomonas bacteria JF2 is 1:

1.

3. A composite bacterial agent, characterized in that: The present invention comprises the Pseudomonas bacteria combination according to claim 1 or 2.

4. The composite bacterial agent according to claim 3, characterized in that The composite bacterial agent includes a bacterial suspension of Pseudomonas bacteria JF1 and a bacterial suspension of Pseudomonas bacteria JF2; the volume ratio of the bacterial suspension of Pseudomonas bacteria JF1 and the bacterial suspension of Pseudomonas bacteria JF2 is 1:1; the OD of the bacterial suspension of Pseudomonas bacteria JF1 and the bacterial suspension of Pseudomonas bacteria JF2 is 600 are 0.8 respectively.

5. Use of the Pseudomonas bacterial combination according to claim 1 or 2 or the composite bacterial agent according to claim 3 or 4 in promoting the growth of Corydalis yanhusuo and / or increasing the total alkaloid content in Corydalis yanhusuo tubers.

6. The use according to claim 5, characterized in that The promoting the growth of Corydalis yanhusuo includes promoting the growth of Corydalis yanhusuo seedlings and / or promoting the growth of Corydalis yanhusuo tubers.

7. The use according to claim 6, characterized in that The method of promoting the growth of Corydalis yanhusuo seedlings includes at least one of 1) and 2): 1) Promote the growth of the above-ground and / or underground parts of Corydalis yanhusuo seedlings; 2) Increase the fresh weight and / or dry weight of the entire Corydalis yanhusuo seedling; The method for promoting the growth of Corydalis tubers includes at least one of (1) and (2): (1) Increase the transverse and / or longitudinal diameter of the tuber of Corydalis yanhusuo; (2) Increase the fresh weight and / or dry weight of Corydalis tubers.

8. The use according to claim 5, characterized in that The promotion of the growth of Corydalis yanhusuo is achieved by the combination of Pseudomonas bacteria to solubilize inorganic phosphorus and / or secrete siderophores.

9. A method for promoting the growth of Corydalis yanhusuo and / or increasing the total alkaloid content in Corydalis yanhusuo tubers, characterized in that: The following steps are involved: The Pseudomonas bacteria combination according to claim 1 or 2 or the composite bacterial agent according to claim 3 or 4 is applied to the Corydalis yanhusuo seedlings.

10. The method according to claim 9, characterized in that The application method includes root irrigation.

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