Paenibacillus mucilaginosus FangSG101 and application thereof

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to paenibacillus mucilaginosus FangSG101 and application thereof. The preservation number of the paenibacillus mucilaginosus FangSG101 is CCTCC NO: M 20232558, and the paenibacillus mucilaginosus FangSG101 can convert insoluble mineral potassium and mineral phosphorus in soil into quick-acting potassium which can be directly absorbed and utilized by plants, has the capability of generating pathogenic bacterium cell wall hydrolase, has a remarkable promotion effect on seed germination and plant growth of tomatoes and cucumbers, and can be applied to actual agricultural production. Through optimization of a culture medium formula and adjustment of a fermentation process, a bacterial powder product with high viable count and high spore rate is obtained.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and in particular relates to a jelly-like Paenibacillus FangSG101 and an application thereof. Background Art

[0002] Paenibacillus mucilaginosus is an important plant rhizosphere bacterium widely distributed in the soil. It was first isolated and purified by scholar Aleksandrov. It is a chemoheterotrophic bacterium that can produce spores. The bacterium can improve plant nutrition and directly promote crop growth through its own metabolism to produce cytokinins, gibberellins, indoleacetic acid and enzymes. It can also indirectly promote crop growth by limiting many diseases caused by various pathogens such as pathogenic Fusarium, Pythium and Rhizoctonia solani and improving soil structure.

[0003] Some jelly-like Bacillus have the functions of solubilizing potassium, dissolving phosphorus and fixing nitrogen. Phosphorus and potassium are the main macronutrients necessary for plant growth and development. They not only participate in the transportation and absorption of nutrients, but also give plants the ability to resist abiotic and biotic stresses. However, more than 90% of potassium exists in insoluble rocks and silicate minerals in the soil in a form that is not directly usable or available to plants, resulting in "potassium-rich but potassium-deficient" soil. Some soil microorganisms can dissolve soil minerals under appropriate conditions, releasing soluble potassium for use by plants. Compared with inorganic chemical fertilizers, using potassium-solubilizing bacteria to decompose soil minerals and release potassium is the method with the broadest application prospects.

[0004] Because it can decompose minerals composed of silicates and aluminosilicates and release soluble elements, it is also called silicate bacteria. It has the ability to decompose and transform silicate minerals and biologically fix nitrogen. In agriculture, jelly-like Bacillus can convert potassium and phosphorus in the soil that are difficult for plants to use into effective potassium and phosphorus for plant use. Under certain conditions, the bacteria form a thick capsule outside the cell, which is sometimes more than 10 times larger than the bacterial body and can form bacterial micelles; at the same time, the viscous extracellular polysaccharides secreted by the bacteria during growth are closely related to its decomposition and transformation of soil minerals; in addition, some organic acids such as citric acid, malic acid, oxalic acid, tartaric acid, etc. are secreted during growth. These acidic substances can dissolve mineral elements when in contact with minerals.

[0005] In agricultural production, Paenibacillus jelly has shown many effects, such as increasing the content of available potassium and available phosphorus in the soil, promoting crop growth, and improving crop yield and quality. It is one of the key and hot spots in the research and development of microbial fertilizers. In addition, the bacteria also have broad applications in mining, metallurgy, sewage flocculation, feed industry and other fields. The development of a strain of Paenibacillus jelly that can release potassium and phosphorus is of great significance for protecting land resources and promoting plant growth. Summary of the invention

[0006] One of the purposes of the present invention is to provide a jelly-like Paenibacillus with potassium-solubilizing and phosphorus-dissolving functions. The jelly-like Paenibacillus FangSG101 involved in the present invention was deposited in the China Center for Type Culture Collection on December 14, 2023, with a deposit number of CCTCC NO: M 20232558, and the deposit address is: Wuhan University, Wuhan, China.

[0007] The jelly-like Bacillus FangSG101 can convert the insoluble mineral potassium and mineral phosphorus in the soil into fast-acting potassium that can be directly absorbed and utilized by plants. It also has the ability to produce pathogen cell wall hydrolases, and has a significant promoting effect on tomato and cucumber seed germination and plant growth, and can be used in actual agricultural production.

[0008] Another object of the present invention is to obtain a bacterial powder product with a high number of viable bacteria and a high spore rate by optimizing the culture medium formula and adjusting the fermentation process. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 This is a picture of the jelly-like Paenibacillus FangSG101 spores under a microscope;

[0010] Figure 2 This is a picture of the colony morphology of the jelly-like Paenibacillus FangSG101 activated by the dilution coating method on a nitrogen-free solid medium after culturing for 48 hours;

[0011] Figure 3 This is a picture of the jelly-like Paenibacillus FangSG101 bacteria under a microscope;

[0012] Figure 4 The graph shows the spore counts of the jelly-like Paenibacillus FangSG101 in Tween dilution solutions of different concentrations;

[0013] Figure 5 The qualitative results of the plate of protease (a), amylase (b) and cellulase (c) produced by Paenibacillus jelly-like FangSG101 are shown;

[0014] Figure 6 (a) is the standard curve of soluble potassium content, and (b) is the comparison chart of potassium solubilization of FangSG101 strain;

[0015] Figure 7 (a) is the standard curve of soluble phosphorus content, and (b) is the comparison chart of phosphorus solubilization of FangSG101 strain;

[0016] Figure 8 The experimental results of FangSG101 fermentation liquid promoting the germination of (a) cucumber seeds and (b) tomato seeds. DETAILED DESCRIPTION

[0017] The present invention is further described in detail below with reference to examples, but the embodiments of the present invention are not limited thereto.

[0018] Example 1 Isolation, Identification and Preservation of Jelly-like Paenibacillus

[0019] 1. Sampling Location

[0020] The sampling locations in this embodiment are soils from farm gardens in Wuhan City, Hubei Province and Luzhou City, Sichuan Province.

[0021] The specific separation and purification steps are as follows:

[0022] The culture medium used for separation was:

[0023] Nitrogen-free culture medium: glucose 10g, K2HPO4 0.2g, MgSO4·7H2O 0.5g, CaCO3 5g, NaCl 0.2g, K2SO4 0.2g, distilled water 1L, pH 7.0-7.2.

[0024] The culture medium used was:

[0025] Nitrogen culture medium: glucose 10g, K2HPO4 0.2g, MgSO4·7H2O 0.5g, CaCO3 5g, NaCl 0.2g, K2SO4 0.2g, yeast powder 0.5g, ammonium sulfate 0.5g, distilled water 1L, pH 7.0-7.2.

[0026] Add 18 g of agar to the above solid culture medium, sterilize under high pressure at 121° C. for 20 min, and culture at 30° C. for 32 h to 48 h.

[0027] 2. Isolation and purification of strains

[0028] (1) Weigh 5 g of soil sample and add it to 45 ml of sterile water. Incubate it in a constant temperature shaker at 30 °C and 200 rpm for 30 min. After standing for 20 min, obtain a sample suspension.

[0029] (2) 1.8 mL of the soil suspension supernatant obtained in step (1) was transferred to a 2 mL cryovial and treated in an 80°C water bath for 10 min;

[0030] (3) Take 100 μL of the sample suspension obtained in step (2) after water bath, and dilute it in a 10-fold gradient to obtain 10 -1 , 10 -2 , 10 -3 times the sample dilution;

[0031] (4) Take the 10 obtained in step (3) respectively -2 , 10 -3 100 μL of the sample dilution was spread on a nitrogen-free solid culture medium plate, and the spread nitrogen-free solid culture medium plate was inverted and cultured in a 30° C. incubator for 48 h; after the culture, colonies grew on the nitrogen-free solid culture medium plate.

[0032] (5) Pick a single colony grown on the nitrogen-free solid culture medium plate of step (4), streak it onto a new nitrogen-free solid culture medium plate, and place the streaked nitrogen-free solid culture medium plate in a constant temperature incubator at 30° C. for 48 h; observe that there is only one colony morphology on the nitrogen-free solid culture medium plate after culturing, and obtain pure colonies, indicating that the separation and purification is complete.

[0033] 3. Strain Identification and Preservation

[0034] The monoclonal culture strain purified in step 2 (5) was cultured in liquid, the cells were collected, the genomic DNA was extracted, and the 16SrDNA fragment was amplified using the universal primers 27F and 1492R described in paragraph 58 of Chinese invention patent ZL202210478937.4. The PCR amplification product was detected by agarose gel electrophoresis, and the PCR product was sequenced. The PCR reaction system: 10× buffer 10μL, 10mM dNTP 2μL, 1μL of each upper and lower primer, 2μL of DNA template, 0.5μL of Taq enzyme, 34μL of ddH2O. Pre-denaturation at 95℃ for 10min; then 94℃ for 30s, 60℃ for 30s, and 72℃ for 1min for a total of 35 cycles, and then extended at 72℃ for 5min. The PCR product was detected by gel electrophoresis and sent to Wuhan Jinkairui Biotechnology Co., Ltd. for sequencing. The identified gene sequence was compared in the NCBI database using the BLAST tool.

[0035] After PCR molecular identification, more than 150 strains of Paenibacillus mucilaginosus were obtained, and the strains were preserved in ultra-low temperature glycerol tubes in the Micro-Kang Probiotics Strain Resource Bank.

[0036] Example 2 Comparative experiment on potassium and phosphorus solubilization, growth and spore production rate of jelly-like Paenibacillus strains

[0037] (1) Starting from the glycerol tube, more than 150 strains of jelly-like Bacillus were activated on solid culture medium, and single colonies were picked and inoculated on solid potassium and phosphate solubilizing plates. The strains were screened by observing the size of the dissolution circle of the culture medium, and 10 potassium and phosphate solubilizing functional strains were selected for re-screening.

[0038] (2) The 10 strains initially screened in step (1) were inoculated into a solid culture medium and placed in a constant temperature incubator at 30°C for 48 hours for activation, and the activated single colonies were picked out and inoculated into a shake flask filled with a seed culture medium for cultivation. The shake flask was filled with 30 mL / 250 mL of liquid and cultured at 30°C and 220 r / min for 14 hours to obtain a seed solution; the seed solution was transferred to a shake flask filled with a fermentation culture medium at a 5% inoculum amount and cultured at 30°C and 220 r / min, and the growth curve was measured, and spore formation was observed by microscopic examination of fixed-point sampling.

[0039] The seed (fermentation) culture medium formula: glucose 10g, K2HPO4 0.2g, MgSO4·7H2O 0.5g, CaCO3 5g, NaCl 0.2g, K2SO4 0.2g, yeast powder 0.5g, ammonium sulfate 0.5g, distilled water 1L, pH 7.0-7.2, sterilized at 121°C for 20min for use.

[0040] More than 150 strains were screened for potassium and phosphorus solubilization functions, and compared with growth rate and spore production. After repeated screening and comprehensive comparison, a jelly-like Paenibacillus strain was selected and named as Paenibacillus jelly-like FangSG101. The strain was derived from the soil of a vegetable garden around Wuhan City, Hubei Province. The number of viable bacteria can reach 2.5x10 after 24 hours of culture. 8 CFU / mL, the spore formation rate can reach more than 80%, and its spore rate and spore production speed are significantly higher than those of other strains. The spore formation rate curve of the top five strains of jelly-like Bacillus with faster spore formation speed is shown in Figure 1 (a) Microscopic examination of spores of FangSG101 strain after 28h of shaking flask culture. Figure 1 (b).

[0041] Morphological comparison of jelly-like Paenibacillus FangSG101 strains:

[0042] (1) The strain FangSG101 was inoculated onto a nitrogen-free solid medium and cultured at 30°C for 48 h. The morphology of the single colonies was observed. Figure 2 As shown, the colony is drop-shaped, colorless and transparent, with a bulge in the middle, neat and smooth edges, a moist and sticky surface, elastic and easy to pull into threads.

[0043] (2) The morphology of strain FangSG101 was observed using an optical microscope. Figure 3 As shown, the bacteria are rod-shaped with a capsule on the surface, and several bacteria can form a floc.

[0044] The jelly-like Bacillus FangSG101 was deposited in the China Center for Type Culture Collection on December 14, 2023, with the deposit number CCTCC NO: M 20232558, and the deposit address: Wuhan University, Wuhan, China.

[0045] Example 3 Preparation of jelly-like Paenibacillus FangSG101 powder and optimization of counting conditions

[0046] 1. Preparation of jelly-like Paenibacillus FangSG101 bacterial powder

[0047] (1) Seed culture medium formula: glucose 10 g, K2HPO4 0.2 g, MgSO4·7H2O 0.5 g, CaCO3 5 g, NaCl 0.2 g, K2SO4 0.2 g, yeast powder 0.5 g, ammonium sulfate 0.5 g, distilled water 1 L, pH 7.0-7.2, sterilized at 121 °C for 20 min.

[0048] (2) Fermentation medium formula: glucose 8 g, corn syrup powder 6 g, K2HPO4 0.2 g, MgSO4·7H2O 0.5 g, CaCO3 3 g, NaCl 0.2 g, K2SO4 0.2 g, yeast powder 1 g, ammonium sulfate 0.5 g, distilled water 1 L, pH 7.0-7.2, sterilized at 121°C for 20 min for use.

[0049] (3) According to step (2) of Example 2 above, a seed solution of Bacillus jelly FangSG101 was prepared, and then transferred to a fermentation tank filled with fermentation medium at a 5% inoculation rate for fermentation culture at 30° C. for about 30 h. When the spore rate reached more than 90%, the fermentation was stopped to obtain a fermentation liquid.

[0050] (4) The fermentation liquid obtained in (3) is centrifuged at 12000 rpm and then the bacterial sludge is collected, and the bacterial powder is obtained by spray drying. The inlet air temperature of the spray drying tower is 158-162°C, the outlet air temperature is 80-84°C, and the vacuum degree in the spray drying tower is -0.01Mpa to -0.02MPa.

[0051] 2. Effects of different Tween concentration dilutions on bacterial powder count results

[0052] Weigh 0.1g of the jelly-like Bacillus FangSG101 powder sample and add it to 9.9g of sterile saline with glass beads containing different concentrations of Tween 80 (0%, 0.05%, 0.1%, 0.5%) diluent, and shake on a shaker at 250rpm for 30 minutes at 25°C to obtain the initial bacterial suspension. Take 2mL of the homogenized initial bacterial suspension into a centrifuge tube and immerse it in a water bath for 80°C water bath treatment for 10 minutes. Take out the bacterial solution after the water bath and cool it to room temperature with ice water to obtain the sample test solution. The test solution is diluted tenfold with sterile saline, and the appropriate dilution is selected for counting to calculate the number of spores in the bacterial powder. The results are as follows Figure 4 As shown in Figure 2, among the four Tween dilutions with different concentrations, the 0% count result was the lowest, which was 3.3×10 10 CFU / g; the highest result was 4.9×10 10 CFU / g, and the spore rate can reach 90%.

[0053] Example 4 Determination of enzyme production capacity of jelly-like Paenibacillus FangSG101

[0054] 1. Detection of protease production ability of jelly-like Paenibacillus FangSG101

[0055] Protease culture medium: 15g skim milk powder, 2g yeast powder, 10g NaCl, 20g agar, sterilize at 115℃ for 20min, pH 7.0.

[0056] Use a toothpick to pick up a small amount of jelly-like Bacillus FangSG101 colony and spot-inoculate it on the protease culture medium. Set up three replicates and culture at 30℃ for 4 days to observe whether a transparent circle appears. The larger the transparent circle, the stronger the ability to produce protease.

[0057] The results are as follows Figure 5 As shown in (a), the jelly-like Bacillus FangSG101 can produce an obvious transparent zone with a diameter of up to 35 mm, and has a strong ability to produce protease.

[0058] 2. Detection of the ability of Paenibacillus jellylike strain FangSG101 to produce amylase

[0059] Amylase culture medium: 5g yeast powder, 10g peptone, 5g NaCl, 18g agar, 10g starch, sterilized at 121℃ for 20min, pH 7.0.

[0060] Use a toothpick to pick up a small amount of jelly-like Bacillus FangSG101 colonies and inoculate them on the amylase culture medium, set up three replicates, culture at 30℃ for 4 days, then cover and stain with dilute iodine solution, rinse with water, and observe whether a transparent circle appears. The larger the transparent circle, the stronger the ability to produce amylase.

[0061] The results are as follows Figure 5 As shown in (b), the gelatinous Bacillus FangSG101 can produce an obvious transparent zone with a diameter of up to 26 mm, and has a good ability to produce amylase.

[0062] 3. Detection of the ability of Bacillus jelly-like FangSG101 to produce cellulase

[0063] Cellulase detection plate: 2-10 g sodium carboxymethyl cellulose, 1.5 g potassium dihydrogen phosphate, 2.5 g peptone, 0.5 g yeast powder, 2.5 g disodium hydrogen phosphate, 1 L water, 15 g agar, pH 7.0, sterilize at 121°C for 20 min.

[0064] Use a toothpick to pick up a small amount of jelly-like Bacillus FangSG101 colony and spot it on the cellulase culture medium. Set up three replicates and culture at 30℃ for 4 days. Add 1g / L Congo red solution to the cultured plate and stain it for 1h. Then soak and wash it twice with 1M NaCl solution for decolorization, each time for 30min. Observe whether there is a cellulose hydrolysis circle around the colony. The larger the hydrolysis circle, the stronger the ability to produce cellulase.

[0065] The results are as follows Figure 5 As shown in (c), the gelatinous Bacillus FangSG101 can produce an obvious transparent zone with a diameter of up to 28 mm, and has a good ability to produce cellulase.

[0066] Hydrolases have the function of hydrolyzing pathogen cell walls and poisoning pathogens. The main components of fungal cell walls are cellulose, glucan, chitin, followed by proteins and lipids. The ability to produce degradative enzymes is an important characteristic of biocontrol and growth-promoting bacteria to inhibit plant pathogens. The results of qualitative enzyme production experiments show that FangSG101 strain has the potential to be developed as a biocontrol strain.

[0067] Example 5 Determination of potassium-solubilizing ability of jelly-like Paenibacillus FangSG101

[0068] (1) A single colony of the jelly-like Paenibacillus sp. FangSG101 was picked from a solid plate and inoculated into a medium containing 10 g of glucose, 0.2 g of K2HPO4, 0.5 g of MgSO4·7H2O, 5 g of CaCO3, 0.2 g of NaCl, 0.2 g of K2SO4, 0.5 g of yeast powder, 0.5 g of ammonium sulfate, and 1 L of distilled water at pH 7.2. The culture was carried out at 30°C and 220 r / min for 14 h to obtain a fermentation broth in the logarithmic growth phase.

[0069] (2) Add 50 mL potassium-dissolving culture medium to a 250 mL conical flask and sterilize at 121°C for 20 min. The culture medium components are 5 g glucose, 0.5 g MgSO4·7H2O, 0.005 g FeCl3, 2 g disodium hydrogen phosphate, 0.1 g CaCO3, 3 g potassium feldspar powder, 1 L distilled water, pH 7.0-7.2. Add 1 mL of the FangSG101 logarithmic growth phase fermentation broth prepared in step (1) of Example 5, and use the inoculated inactivated fermentation broth as a control. Each treatment is repeated 3 times. After culturing at 30°C and 220 r / min for 14 days, the culture is terminated. The culture broth is centrifuged at 10,000 r / min for 5 min, the supernatant is collected, 1 mL 6% H2O2 is added for digestion for 1 h, and then centrifuged again to obtain the supernatant. With reference to the relevant contents of NYT 889-2004, the industry standard for the determination of soil fast-acting potassium and slow-acting potassium content, the water-soluble potassium content in the supernatant was determined by flame spectrophotometry, and compared with the control treatment, the potassium-solubilizing capacity of FangSG101 strain was calculated.

[0070] Potassium release amount = water-soluble potassium ion content of the experimental group - water-soluble potassium ion content of the control group

[0071] like Figure 6 (a) shows the standard curve of soluble potassium content determined by flame photometer; Figure 6 As shown in (b), the results showed that after 14 days of cultivation, the average effective potassium content of the FangSG101 treatment group was 56.01±3.71 mg / L, and the average effective potassium content of the CK group was 22.41±0.69 mg / L. The FangSG101 treatment group increased by 33.6 mg / L compared with the control, indicating that the FangSG101 strain has a certain potassium-solubilizing ability.

[0072] Example 6 Determination of the phosphate solubilization ability of the jelly-like Paenibacillus FangSG101

[0073] The preparation process of the jelly-like Paenibacillus FangSG101 bacterial suspension is the same as step (1) of Example 5.

[0074] 50 mL of phosphate solubilization medium was added to a 250 mL conical flask. The medium components were 10 g glucose, 0.5 g ammonium sulfate, 0.2 g NaCl, 0.2 g KCl, 0.1 g MgSO4·7H2O, 0.005 g MnSO4·7H2O, 0.005 g FeSO4·7H2O, and 0.5 g yeast powder, with a pH of 7.0-7.2. 1 mL of bacterial suspension was inoculated as the experimental group, and the inactivated bacterial suspension was inoculated as the control. Each treatment was repeated 3 times. After 14 days of cultivation at 30°C and 220 r / min, the culture was terminated, the culture solution was centrifuged at 10000 r / min for 5 min, and the supernatant was collected. Referring to the relevant content of the environmental protection standard HJ 632-2011 for the determination of total phosphorus in soil, the content of soluble phosphorus in the fermentation broth of the strain was detected by molybdenum antimony anti-spectrophotometry, and compared with the control treatment to calculate the amount of phosphorus solubilization.

[0075] Phosphorus solubilization amount = effective phosphorus content of experimental group - effective phosphorus content of control group

[0076] like Figure 7 (a) shows the standard curve of soluble phosphorus content determined by molybdenum antimony anti-spectrophotometry; Figure 7 (b) The results showed that after culturing for 14 days, the average available phosphorus content in the FangSG101 treatment group was 130.23±2.95 mg / L, and the average available phosphorus content in the CK group was 66.13±4.88 mg / L. The FangSG101 treatment group increased by 63.87 mg / L compared with the control, indicating that the FangSG101 strain has a certain ability to solubilize phosphorus.

[0077] Example 7 Determination of the ability of Paenibacillus jellylike FangSG101 to promote seed germination

[0078] Refer to GB / T 3543.4-1995 Crop Seed Inspection Procedure (Germination Test), select full cucumber and tomato seeds, soak them in 75% alcohol for 30 seconds, then rinse them with sterile water until there is no alcohol residue; then soak them in sodium hypochlorite solution for 20 minutes, and then rinse them with sterile water several times to complete seed disinfection. Place a sterile filter paper in a culture dish (9 cm in diameter), and evenly sprinkle the sterilized seeds (50 tomato seeds / dish, 25 cucumber seeds / dish) on the filter paper in an ultra-clean workbench, add two different treatment solutions, sterile water (control group) and FangSG101 fermentation solution that has been cultured for 48 hours (treatment group), 100 seeds for each treatment, and repeat 3 times. Place them in a 30℃ constant temperature incubator to moisturize and germinate, and count the seed germination after 7 days, and measure the germination length with a ruler.

[0079] Seed germination rate (%) = number of germinated seeds / number of sown seeds × 100

[0080] Table 1 Effects of FangSG101 fermentation liquid on cucumber and tomato seed germination

[0081]

[0082] Note: The data in the table are mean ± standard error. Lowercase letters indicate significant differences in germination rate between the control group and the treatment group (P<0.05).

[0083] The cucumber and tomato seeds were treated with the 48h fermentation liquid of FangSG101 strain, and the germination rates of the seeds in the treatment group and the sterile water control group are shown in Table 1. Among them, the germination rate of cucumber seeds in the FangSG101 fermentation liquid treatment group was 91.33±1.70%, and that in the sterile water control group was 84.33±1.25%; the germination rate of tomato seeds in the FangSG101 fermentation liquid treatment group was 87.67±0.47%, and that in the sterile water control group was 78.33±2.05%; the results showed that there were significant differences between the FangSG101 fermentation liquid treatment group and the sterile water control group, and the strain FangSG101 could promote seed germination.

[0084] In addition, the growth potential of cucumber seeds in the FangSG101 strain treatment group was generally higher than that in the sterile water control group. The example results are as follows Figure 8 (a) shows that the growth potential of tomato seeds in the FangSG101 strain treatment group is generally higher than that in the sterile water control group. The example results are shown in Figure 8 (b) as shown.

[0085] Example 8 Determination of the ability of Paenibacillus jelly-like FangSG101 to promote the growth of cucumber seedlings

[0086] The cucumber seeds that germinated and grew uniformly in Example 7 were selected and transferred to sterilized nutrient soil (containing a certain proportion of peat soil, vermiculite, perlite, etc.) potting pots (diameter 18.5 cm, height 16.5 cm, bottom diameter 13 cm), and three seedlings were inoculated in each pot. 20 mL of 1.0×10 8 The fermentation broth with CFU / mL cultured for 48 hours was used as the treatment group, and sterile water was inoculated as the blank control group, and then the surface was covered with 1-2 cm of sterilized nutrient soil.

[0087] The sterile water control group and FangSG101 fermentation liquid treatment group used 6 seedling pots, each with 18 cucumber seedlings. After the cucumber seedlings were cultured in a greenhouse with a temperature of 25±1℃, a relative humidity of 55%±5%, and a photoperiod of 14 hours of light / 10 hours of darkness for 28 days, the root length, plant height, plant stem diameter, plant fresh weight and plant dry weight of the plant were measured to evaluate the growth-promoting effect of FangSG101 strain on cucumber.

[0088] Table 2 The promoting effect of FangSG101 fermentation liquid on cucumber growth (28 days)

[0089]

[0090] Note: The data in the table are mean ± standard error. Lowercase letters in the same column indicate significant differences in seedling growth between the control group and the treatment group (P<0.05).

[0091] The results of the greenhouse potted test data are shown in Table 2. The root length, stem diameter, plant height, fresh weight and dry weight of cucumber seedlings in the soil of the group treated with the fermentation liquid of Bacillus jelly FangSG101 increased by 44.95%, 49.36%, 43.48%, 68.45% and 60% respectively compared with the sterile water control group, indicating that Bacillus jelly FangSG101 can significantly promote the growth of cucumber plants.

[0092] Example 9 Determination of the ability of Paenibacillus jelly-like FangSG101 to promote the growth of tomato seedlings

[0093] The tomato seeds that germinated and grew evenly in Example 7 were selected and transferred to sterilized nutrient soil (containing a certain proportion of peat soil, vermiculite, perlite, etc.) potting pots (diameter 18.5 cm, height 16.5 cm, bottom diameter 13 cm), and three seedlings were inoculated in each pot. 20 mL of 1.0×10 8 The fermentation broth with CFU / mL cultured for 48 hours was used as the treatment group, and sterile water was inoculated as the blank control group, and then the surface was covered with 1-2 cm of sterilized nutrient soil.

[0094] Six seedling pots were used in the sterile water control group and the FangSG101 fermentation liquid treatment group, with 18 tomato seedlings in each group. After the tomato seedlings were cultured in a greenhouse with a temperature of 25±1℃, a relative humidity of 55%±5%, and a photoperiod of 14 hours of light / 10 hours of darkness for 28 days, the root length, plant height, plant stem diameter, plant fresh weight and plant dry weight of the plant were measured to evaluate the growth-promoting effect of FangSG101 strain on tomato.

[0095] Table 3 The promoting effect of FangSG101 fermentation liquid on tomato growth (28 days)

[0096]

[0097] Note: The data in the table are mean ± standard error. Lowercase letters in the same column indicate significant differences in seedling growth between the control group and the treatment group (P<0.05).

[0098] The results of the greenhouse potted test data are shown in Table 3. The root length, stem diameter, plant height, fresh weight and dry weight of tomato seedlings in the soil of the group treated with the fermentation liquid of Bacillus jelly FangSG101 increased by 30.32%, 26.17%, 15.19%, 26.83% and 22.73% respectively compared with the sterile water control group, indicating that Bacillus jelly FangSG101 can significantly promote the growth of tomato plants.

Claims

1. Paenibacillus jelly-like strain FangSG101 is deposited in China Center for Type Culture Collection with the deposit number of CCTCCNO: M 20232558.

2. The jelly-like Paenibacillus FangSG101 described in claim 1 is used as a biocontrol bacterium.

3. The use according to claim 2, characterized in that: The jelly-like Paenibacillus FangSG101 has the ability to produce protease.

4. The use according to claim 2, characterized in that: The jelly-like Paenibacillus FangSG101 has the ability to produce cellulase.

5. The use according to claim 2, characterized in that: The gelatinous Paenibacillus FangSG101 has the ability to produce amylase.

6. The jelly-like Paenibacillus FangSG101 of claim 1 is used for dissolving potassium and phosphorus in soil.

7. The method for culturing the jelly-like Paenibacillus FangSG101 according to claim 1, characterized in that: (1) Prepare the seed culture medium formula: glucose 10 g, K2HPO4 0.2 g, MgSO4·7H2O 0.5 g, CaCO3 5 g, NaCl 0.2 g, K2SO4 0.2 g, yeast powder 0.5 g, ammonium sulfate 0.5 g, distilled water 1 L, pH 7.0-7.2, sterilize at 121°C for 20 min for later use; (2) Prepare the fermentation medium formula: glucose 8 g, corn syrup powder 6 g, K2HPO4 0.2 g, MgSO4·7H2O 0.5 g, CaCO3 3 g, NaCl 0.2 g, K2SO4 0.2 g, yeast powder 1 g, ammonium sulfate 0.5 g, distilled water 1 L, pH 7.0-7.2, sterilize at 121°C for 20 min for use; (3) A seed solution of Paenibacillus jelly-like FangSG101 was prepared, and then transferred to a fermentation tank filled with fermentation medium at a 5wt% inoculation rate for fermentation culture at 30°C for about 30 hours. When the spore rate reached more than 90%, the fermentation was stopped to obtain a fermentation liquid.

8. The jelly-like Paenibacillus FangSG101 of claim 1 is used as a plant growth additive or used for preparing a plant growth additive.

9. The use according to claim 8, characterized in that: The plant is cucumber or tomato.

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