A strain of Bacillus zanthoxylum HN90 and its synergist and its application in preparing biological organic fertilizer

Abstract

The present invention relates to the field of microbial technology, and in particular to a strain of Bacillus zanthoxylum HN90 and a synergist thereof and an application thereof in preparing a biological organic fertilizer. The Bacillus zanthoxylum HN90 of the present invention is a probiotic selected by an applicant. After testing, the strain has multiple growth-promoting characteristics, specifically, it can have a higher IAA production capacity, phosphorus dissolving, phosphorus solubilizing and nitrogen fixing functions. After seedling raising experiments, it is also found that the strain prepared into a hydroponic solution can effectively promote the growth of rice, indicating that the Bacillus zanthoxylum HN90 of the present invention can still promote the growth of rice under flooding conditions. In addition, in the growth-promoting experiment of biostimulants, the applicant also found that gamma-aminobutyric acid (GABA) has a good synergistic effect on Bacillus zanthoxylum HN90; in the seedling raising experiment, it is further found that when HN90 and GABA are used simultaneously, the seedlings grow more robustly, without yellow leaves, and the amount of nitrogen fertilizer can be reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of microorganisms, and in particular to a Bacillus zanthoxylum HN90 and a synergist thereof, and application of the same in preparing biological organic fertilizers. Background Art

[0002] As one of the three major staple foods, rice is of great significance to ensuring food security. At present, agricultural production increases soil nutrients by increasing the use of chemical fertilizers. However, the large-scale application of chemical fertilizers will lead to soil acidification and compaction, and imbalance of soil structure and microbial structure. Long-term and unreasonable application will cause irreversible damage to the ecological environment.

[0003] Bacillus is a common genus of beneficial bacteria that promotes plant growth in the rhizosphere. It possesses strong stress resistance, a broad antibacterial spectrum, is easy to survive, and has low propagation costs. It promotes plant growth directly and indirectly through secretion of phytohormones, phosphate and potassium solubilization, siderophore secretion, plant resistance induction, and pathogen inhibition. Bacillus species are numerous, highly resilient, and have minimal impact on humans and the environment, making it a key bacterial species in the study of plant beneficial bacteria. However, common Bacillus products on the market often exhibit reduced growth-promoting properties under flooding conditions, likely due to the ability of the functional strains to colonize in such conditions. Biostimulants are substances that, when applied to seeds, crops, or the rhizosphere, stimulate their natural growth processes, enhancing nutrient absorption, fertilizer utilization efficiency, and tolerance to abiotic stresses, thereby improving crop quality and yield. They are categorized into microbial and non-microbial biostimulants. Combining functional microorganisms with non-microbial biostimulants to form composite biostimulants significantly enhances their effectiveness. However, different non-microbial biostimulants have different growth-promoting effects on plants and microorganisms. Therefore, it is necessary to screen out special rhizospheric bacteria that can be used for rice growth according to the special growth environment of rice. In addition to having a good growth-promoting effect, the rhizospheric bacteria should also have a good colonization ability in the rice rhizosphere under rice hydroponic conditions, thereby playing a significant role in promoting rice growth under flooding conditions. In addition, synergists for specific single strains of functional bacteria should be developed to achieve the effect of simultaneously improving rice yield and quality. Summary of the Invention

[0004] In view of the above, it is necessary to screen out special rhizospheric bacteria that can be used for rice growth according to the special growth environment of rice. In addition to having a good growth-promoting effect, the rhizospheric bacteria should also have a good colonization ability in the rice rhizosphere under rice hydroponic conditions, thereby playing a significant role in promoting rice growth under flooding conditions. In addition, synergists for the functional bacteria should be developed for specific single strains, which can simultaneously improve rice yield and quality.

[0005] In order to achieve the above purpose, the present invention screened out a new strain: Bacillus zanthoxylum (Bacillus zanthoxyli ) strain HN90, which is classified as: Bacillus zanthoxyli HN90, named in Chinese as Bacillus zanthoxylum HN90, with a deposit number of CCTCC NO: M20241255, is deposited in the China Center for Type Culture Collection, Wuhan University, Wuhan, China. The detailed address is: No. 299, Bayi Road, Wuchang District, Wuhan University, opposite the First Affiliated Primary School, on the campus of Wuhan University. The deposit date is June 17, 2024.

[0006] The present invention also includes the Zanthoxylum bungeanum Bacillus ( Bacillus zanthoxyli ) strain HN90.

[0007] The present invention also includes the Zanthoxylum bungeanum Bacillus ( Bacillus zanthoxyli ) Application of strain HN90 or the bacterial agent in the preparation of bio-organic fertilizer and / or water-based fertilizer.

[0008] The present invention also includes the Zanthoxylum bungeanum Bacillus ( Bacillus zanthoxyli ) Application of strain HN90 or the bacterial agent in promoting rice growth.

[0009] The present invention also includes the Zanthoxylum bungeanum Bacillus ( Bacillus zanthoxyli ) Application of strain HN90 or the bacterial agent in producing IAA, solubilizing phosphate, dissolving phosphate and / or fixing nitrogen.

[0010] The present invention also includes a method for preparing water fertilizer using the Bacillus zanthoxyli strain HN90 or the bacterial agent, the method comprising: preparing the Bacillus zanthoxyli HN90 into a suspension and then diluting the suspension with sterile water to an OD of 600 =1.

[0011] The present invention also includes a synergist for the growth of the Bacillus zanthoxyli strain HN90, wherein the synergist is gamma-aminobutyric acid (GABA); and the concentration of the GABA is 1 g / L-3 g / L.

[0012] Furthermore, the method comprises inoculating a bacterial liquid of Bacillus zanthoxylum HN90 into ordinary organic fertilizer for fermentation to obtain a bio-organic fertilizer. The commercial organic fertilizers in this application are: organic fertilizer (Organic Fertilizer 1) produced by Jiangsu Hexi Biotechnology Co., Ltd. using traditional Chinese medicine residue and bean dregs as primary raw materials; organic fertilizer (Organic Fertilizer 2) produced by Changzhou Hongbao Biotechnology Co., Ltd. using traditional Chinese medicine residue and straw as primary raw materials; or organic fertilizer (Organic Fertilizer 3) produced by Nanjing Ningliang Bioengineering Co., Ltd. using mushroom residue as primary raw materials.

[0013] The present invention also includes a method for using the Bacillus zanthoxyli strain HN90 or the bacterial agent to reduce the amount of chemical fertilizer used in rice cultivation. The method comprises: when planting rice, reducing the amount of nitrogen used in chemical fertilizer by 20% compared with the amount of nitrogen used in conventional chemical fertilizers; and then applying 10 L / mu to 20 L / mu of a compound solution of Bacillus zanthoxyli HN90 and GABA in the field during the tillering period; the amount of nitrogen used in conventional chemical fertilizers is 300 kg / ha.

[0014] Furthermore, the content of Bacillus zanthoxylum HN90 is 1×10 9 CFU / mL, and the GABA content is 1 g / L.

[0015] The present invention has the following beneficial effects: The Zanthoxylum Bacillus HN90 of the present invention is a probiotic strain independently selected by the applicant. Testing has shown that this strain has multiple growth-promoting properties, specifically a high IAA production capacity, as well as excellent phosphate solubilization, phosphate dissolution, and nitrogen fixation capabilities. Seedling cultivation experiments also revealed that the strain, when prepared into a hydroponic solution, effectively promoted rice growth, whereas the hydroponic solution of the same genus strain SQR9 did not promote rice growth. This demonstrates that the Zanthoxylum Bacillus HN90 of the present invention can still promote rice growth under flooding conditions. Furthermore, in biostimulant growth-promoting experiments, the applicant also found that GABA promotes the growth of both Zanthoxylum Bacillus HN90 and rice. Field trials have shown that the application of HN90 and GABA as a whole with organic fertilizer can reduce nitrogen fertilizer usage without reducing yield. This demonstrates that the Zanthoxylum Bacillus HN90 of the present invention and GABA are a good fertilizer-reducing agent and a good hydroponic fertilizer for rice. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a plate image of strain HN90.

[0017] Figure 2 This is the phylogenetic tree of strain HN90.

[0018] Figure 3 This is the phosphate solubilization circle diagram of strain HN90.

[0019] Figure 4 This is the phosphate circle diagram of strain HN90.

[0020] Figure 5 This is the nitrogen fixation circle diagram of strain HN90.

[0021] Figure 6 This is the pH effect diagram of strain HN90.

[0022] Figure 7 This figure shows the growth-promoting effect of strain HN90 on rice.

[0023] Figure 8 This diagram shows the effect of strain HN90 on rice roots.

[0024] Figure 9 This figure shows the effect of GABA on the growth of strain HN90.

[0025] Figure 10 This figure shows the effect of AOS on the growth of strain HN90.

[0026] Figure 11 This figure shows the effect of PGA on the growth of strain HN90.

[0027] Figure 12 This figure shows the effect of different concentrations of GABA on the growth of rice seedlings.

[0028] Figure 13 Schematic diagram of rice seedling cultivation in the experimental group and CK group treated with the compound solution of GABA and HN90.

[0029] Figure 14 Schematic diagram of the rice root conditions in the experimental group and CK group treated with the GABA and HN90 compound solution. DETAILED DESCRIPTION

[0030] All features disclosed in this specification, or all steps in the disclosed methods or processes, except mutually exclusive features and / or steps, can be combined in any manner.

[0031] Any feature disclosed in this specification (including any accompanying claims and abstract), unless otherwise stated, is merely an example of a series of equivalent or similar features. Example 1

[0032] The probiotics used in this example are Bacillus zanthoxylum ( Bacillus zanthoxyli ) strain HN90, which is classified as: Bacillus zanthoxyli HN90, the Chinese classification name is: Bacillus zanthoxylum HN90, the preservation number is CCTCC NO: M20241255; the strain is deposited in the China Center for Type Culture Collection, address: Wuhan University, Wuhan, China, and the preservation date is June 17, 2024.

[0033] 1. Molecular identification of strains: Based on 16S sequence comparison and phylogenetic tree analysis, the species of strain HN90 was analyzed. The phylogenetic tree is shown in Figure 2 , as can be seen from the figure: HN90 strain and Bacillus Belong to one category, Bacillus zanthoxyli species have a high homology; therefore, the HN90 strain was identified as Bacillus zanthoxyli kind.

[0034] 2. Morphological identification Morphological identification such as Figure 1 As shown: The colony morphology of HN90 strain inoculated in LB medium is characterized by white colonies, rough edges, round colonies, and wrinkles in the middle. Bacillus zanthoxyli ). Comprehensive molecular identification classified the strain as Bacillus zanthoxylum. Example 2

[0035] The growth promotion test of Zanthoxylum bungeanum strain HN90 is as follows.

[0036] 1. Determination of IAA production capacity: IAA production was determined using the Salkowski colorimetric method (Glickmann and Dessaux, 1995). 1% inoculum of Trichoderma brevis TB2 spores was added to a 200 mg·L -1 The culture medium was TSB with tryptophan, and the culture was shaken at 170 rpm at 28°C in the dark for 5 days. -1 Centrifuge at 4°C for 10 minutes. Take 100 μL of the fermentation supernatant and add Salkowski colorimetric solution at a 1:1 ratio. Incubate in the dark for 30 minutes. If the reaction solution turns red, it indicates the presence of IAA analogs. Darker colors indicate higher IAA content. Determine the OD530 value using a microplate reader and calculate the IAA production of the strain using the corresponding standard curve.

[0037] The standard curve was prepared using IAA standard (Sigma). The concentrations of the prepared IAA solutions were 0, 10, 20, 30, 40, and 50 mg / L, respectively. 100 μL of each gradient solution was aspirated, 100 μL of colorimetric solution was added, and the reaction was carried out in the dark for 30 minutes before measuring OD530 nm. TSB blank culture medium with colorimetric solution added was used as a control, and three replicates were set up to draw a standard curve. The ability of the new strain HN90 of the present application to produce IAA on the second day was determined by the above method. At the same time, the applicant also determined the IAA production of Bacillus Velezii SQR9 (CGMCC No. 5808), a strain of the same genus with strong disease resistance and growth promotion ability, which was isolated from the cucumber rhizosphere by the applicant's team. The test results of the two strains are shown in Table 1.

[0038]

[0039] As shown in Table 1, the IAA production of Bacillus zanthoxylum HN90 reached 34.9 mg / L on the second day, indicating that the new strain HN90 has a higher IAA production ability, while the IAA production of strain SQR9 reached 37.0 mg / L on the second day, indicating that the IAA production of the same genus strain SQR9 is slightly higher than that of the new strain Bacillus zanthoxylum HN90. In theory, both have the potential to promote plant growth.

[0040] 2. Determination of phosphate solubilization, phosphate dissolving and nitrogen fixation ability: 10 μL of HN90 seed solution was inoculated on a modified Ashby (nitrogen-free) agar plate, an inorganic phosphorus bacterial culture plate and an organic phosphorus bacterial culture plate, respectively. The plates were incubated at 30°C for 7 days to observe whether hydrolysis circles were generated. Figure 3 As shown in the figure: It can be seen that strain HN90 can produce a dissolution circle and dissolve phosphorus. Figure 4 As shown in the figure: It can be seen that the strain HN90 can produce a dissolution zone, and the nitrogen fixation ability effect is as follows Figure 5 As shown: the lower row in the figure is other strains in the screening process, and the upper row is the new strain HN90 of this application; it can be seen from the figure that strain HN90 can produce a dissolution circle, which shows that strain HN90 has strong phosphate solubilization, phosphate dissolving and nitrogen fixation functions, and can be used as a raw material for crop fertilizer. Example 3

[0041] This example studies the pH tolerance of Bacillus zanthoxylum HN90. The specific method is as follows: 1% inoculum of Bacillus zanthoxylum HN90 was inoculated into TSB medium with pH values ​​of 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, and 9.0. The effect of different initial pH on the growth of Bacillus zanthoxylum HN90 was determined. The test results are shown in Figure 2. Figure 6 As shown in the figure, it can be seen that Bacillus zanthoxylum HN90 can grow in TSB culture medium at 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, and 9.0, indicating that Bacillus zanthoxylum HN90 has the ability to grow in a wide range of pH. Example 4

[0042] This example studies the growth-promoting effect of Bacillus zanthoxylum HN90 on rice in a hydroponic environment.

[0043] 1. Test materials: Bacillus zanthoxylum HN90, Bacillus velezensis SQR9, and rice seeds (Yliangyou-689).

[0044] 2. Preparation of bacterial hydroponic solution: Inoculate Bacillus zanthoxylum HN90 strain or Bacillus velezensis SQR9 on TSB medium, culture at 28℃ for 24 h, and adjust the OD value to 0. 600 =1, to prepare the hydroponic solution.

[0045] 3. Experimental Treatments: Rice seeds were pre-soaked in sterile deionized water for one day, then soaked in 75% alcohol for 1 minute, rinsed 3–5 times with sterile water, and then soaked in 2% sodium hypochlorite solution for 30 minutes, followed by 5 times with sterile water. Sterilized rice seeds were evenly spread on sterilized filter paper in the dark and germinated until white. White seeds were selected for hydroponics, with one leaf and one heart. The seeds were then placed in hydroponic culture solutions for different groups and inoculated with bacteria. Group CK served as a blank control group, treated only with sterile water and not inoculated. The experimental results are shown in Table 2.

[0046]

[0047] Note: The same lowercase letters in the same column indicate no significant difference ( P>0.05 ), different lowercase letters indicate significant differences ( P<0.05 ).

[0048] As can be seen from Table 2, Bacillus zanthoxylum HN90 can still promote rice growth after being prepared into hydroponic solution, and the fresh weight and dry weight of rice can be significantly improved compared with the control group and SQR9 experimental group. The experimental results also show that although the same genus strain SQR9 has a high IAA production ability and has been proven to have strong disease resistance and growth-promoting ability, it cannot significantly promote rice growth after being prepared into hydroponic solution. Its fresh weight and dry weight are not significantly different from those of the control group, and even the fresh weight of the aboveground part is significantly lower than that of the CK group.

[0049] In addition, from Figure 7 and Figure 8 It can also be seen that the growth and root growth of rice added with the hydroponic solution of Bacillus zanthoxylum HN90 are better than those of the blank control group (CK) and the same genus Bacillus group (SQR9); the above experimental results show that not all reported growth-promoting strains can effectively promote the growth of all plants. The experiments of this application have proved that the control Bacillus SQR9 cannot promote the growth of rice. After Bacillus zanthoxylum HN90 is prepared into a hydroponic solution, it has a good growth-promoting ability for rice and is an excellent rice hydroponic agent. Example 5

[0050] This example studies the effects of stimulants on the growth of Bacillus zanthoxylum HN90 and rice: In a large number of experiments conducted by the applicant, it was found that some biostimulants have a specific growth-stimulating effect on the strain. In order to improve the growth and adaptability of the strain, this example studies the specific effects of the biostimulants brown algal oligosaccharides (AOS), γ-aminobutyric acid (GABA) and polyglutamic acid (PGA) on the growth of Bacillus zanthoxylum HN90. The specific scheme is as follows.

[0051] 1. Effect of biostimulants on the biomass of Bacillus zanthoxylum HN90: Three biostimulants AOS, GABA and PGA were prepared with deionized water to 11 gradient concentration solutions of 0.3 g / L, 0.6 g / L to 3 g / L. 10% of the solution was added to 10% TSB and cultured for 12 h to measure the OD 600 , to explore the effects of different concentrations of different biostimulants on the biomass of rice rhizosphere growth-promoting bacteria. Figure 9 、 Figure 10 and Figure 11 As shown, of the three stimulants, only GABA significantly promoted the growth of Bacillus zanthoxylum HN90, while AOS and PGA showed no growth-promoting effect. The results also showed that higher GABA concentrations increased the biomass of Bacillus zanthoxylum HN90 and exhibited a more pronounced growth-promoting effect. This suggests that GABA is feasible as a synergist for Bacillus zanthoxylum HN90.

[0052] 2. Effect of biostimulant GABA on rice growth: Biostimulant GABA was prepared with deionized water to 0.025 g / L, 0.05 g / L, 0.1 g / L, 0.2 g / L, and 0.4 g / L solutions (as shown in Table 3). 50 mL of solution was inoculated into each pot after germination and whitening. 50 mL of solution was inoculated into each pot again after 14 days. The rice was harvested after 30 days. The results are shown in the figure below. Figure 12 shown.

[0053]

[0054] The results of root irrigation of rice with the above concentration of GABA are as follows: Figure 12 As shown in the figure, it can be seen that different concentrations of GABA promote the growth of rice seedlings. When the GABA content is G5 (0.4g / L), it has the most significant growth-promoting effect on rice growth. Example 6

[0055] This embodiment is the preparation of Bacillus zanthoxylum HN90 bio-organic fertilizer: by using an acid-hydrolyzed amino acid and three kinds of decomposed plant-derived organic fertilizers and adding Bacillus zanthoxylum HN90 for secondary fermentation, a rice-specific bio-organic fertilizer product rich in a large number of functional microorganisms is produced, as shown in Table 4. The specific scheme is as follows.

[0056] Acid hydrolyzed amino acids were added to the decomposed organic fertilizer at 2% (w / w) (dry weight). After thorough stirring, the Zanthoxylum bungeanum HN90 bacterial solution was added at a concentration of 0.1 mL·g -1 The seeding density is 6×10 7 CFU·mL -1The fermentation liquid of Bacillus zanthoxylum HN90 was inoculated into three kinds of plant-derived organic fertilizers from different manufacturers in Table 3, so that the number of viable bacteria in the organic fertilizers reached 6×10 6 CFU·g -1 , carry out secondary fermentation solid production of bio-organic fertilizer, pile it at room temperature, turn it over once a day, after 7 days, take 5g of bio-organic fertilizer, add 45ml of deionized water, shake it at 28℃ for 30min, then let it stand for 10min, dilute the supernatant and apply it to determine the number of viable bacteria.

[0057] The above preparation scheme includes three types of plant-based organic fertilizers, namely, the organic fertilizer (organic fertilizer 1) produced by Jiangsu Hexi Biotechnology Co., Ltd. using Chinese medicine residue and bean dregs as the main raw materials, the organic fertilizer (organic fertilizer 2) produced by Changzhou Hongbao Biotechnology Co., Ltd. using Chinese medicine residue and straw as the main raw materials, or the organic fertilizer (organic fertilizer 3) produced by Nanjing Ningliang Bioengineering Co., Ltd. using mushroom residue as the main raw material.

[0058]

[0059] As can be seen from Table 4, after 7 days, the number of viable bacteria in organic fertilizer 1, organic fertilizer 2 and organic fertilizer 3 was higher than the initial number, indicating that Bacillus HN90 can be further multiplied and strengthened in the above organic fertilizers, and the fertilizer effect will be better. Example 7

[0060] This example studies the effect of a combination of Bacillus zanthoxylum HN90 and GABA on rice yield in the field. Specifically, the following experimental scheme was used to treat rice: CK: no fertilization; TF: local customary fertilization rate, with a nitrogen fertilizer rate of 300 kg / ha; OF1, OF2, and OF3 were experimental groups with different dosages of the compound solution. During the planting period, the nitrogen fertilizer rate was reduced by 20%, i.e., 240 kg / ha. The OF experimental group, while reducing the nitrogen fertilizer rate, applied a compound solution of Bacillus zanthoxylum HN90 and GABA in the field during the tillering period (the HN90 content of the compound solution was 1×10 9 CFU / mL, GABA content is 1g / L), wherein, the compound liquid applied to the OF1 experimental group is 10L / mu, the compound liquid applied to the OF2 experimental group is 15L / mu, and the compound liquid applied to the OF3 experimental group is 20L / mu. The rice is harvested after 3 months. In this embodiment, the effect of the obtained Zanthoxylum bungeanum HN90 and GABA compound on rice yield is shown in Table 5.

[0061]

[0062] Note: The same lowercase letters in the same column indicate no significant difference ( P>0.05 ), different lowercase letters indicate significant differences ( P<0.05 ).

[0063] As can be seen from Table 5, compared with the TF farmers' habit, after applying the compound solution of Bacillus zanthoxylum HN90 and GABA, although the three experimental groups of OF1, OF2 and OF3 reduced the amount of chemical fertilizer (the amount of chemical fertilizer was reduced by 20% compared with TF), their number of effective ears, number of grains per ear, and yield were not significantly different from those of the TF (farmers' habit of fertilization) experimental group, but were significantly higher than the control group CK without fertilizer application. This shows that the compound solution treatment can significantly increase rice yield, achieve reduced chemical fertilizer application, and is an economical and effective fertilization scheme. Example 8

[0064] This example studies the effect of the compound of Zanthoxylum bungeanum HN90 and GABA on the growth of seedlings: The specific scheme is as follows: a commercial seedling raising matrix (CK) and a compound solution of Zanthoxylum bungeanum HN90 and GABA are added to the matrix. The commercial matrix is ​​purchased from Huaian Chaimihe Agricultural Science and Technology Co., Ltd. The seedling raising experiment was carried out on June 16, 2024. Hard plastic seedling raising trays (specifications 58cm×28cm×3cm) were used for seedling raising. Three trays were used for each treatment. The weight of the matrix in the seedling tray was 4.5kg (dry weight), and the dry weight of rice seeds sown in each tray was 120g. Before sowing rice, the seeds were mixed with seed dressing, naturally dried, and then weighed and sown. Before sowing, the matrix was irrigated with water, and a special seeder was used to evenly sow, cover with soil, and water. After 28 days of seedling raising according to the above scheme, the growth of its roots is as follows Figure 13 and Figure 14 As shown, from Figure 13 It can be seen that the roots of the CK group, which was not treated with the compound solution of Bacillus zanthoxylum HN90 and GABA, were burnt yellow, while the roots of the experimental group, which was treated with the compound solution of Bacillus zanthoxylum HN90 and GABA, were green and in good growth condition. Figure 14 It can be seen that the roots of the CK group, which was not applied with the compound solution of Bacillus zanthoxylum HN90 and GABA, were less distributed outside the seedling tray and the roots were black, while the roots of the experimental group, which was applied with the compound solution of Bacillus zanthoxylum HN90 and GABA, were well-developed, whiter in color, and more distributed outside the seedling tray.

[0065] In summary, the Zanthoxylum bungeanum HN90 screened by the applicant of the present invention has multiple growth-promoting properties (producing the hormone IAA, solubilizing phosphate, dissolving phosphate, and fixing nitrogen), has a wide pH adaptability range, and can promote the growth of rice under flooding conditions. In addition, through research on biotin, it was found that GABA has a promoting effect on the growth of both Zanthoxylum bungeanum HN90 and rice; field experiments have shown that after compounding Zanthoxylum bungeanum HN90 and GABA, the amount of nitrogen fertilizer used can be reduced without reducing yield, effectively achieving the reduction of chemical fertilizers.

[0066] The above-described embodiments merely illustrate several embodiments of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art would be able to make various modifications and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

Claims

1. Bacillus prickly ash Bacillus zanthoxyli The strain HN90 has a deposit number of CCTCC NO: M20241255.

2. comprising the Zanthoxylum bungeanum described in claim 1 Bacillus zanthoxyli Inoculum of strain HN90.

3. Bacillus zanthoxylum as claimed in claim 1 Bacillus zanthoxyli Use of strain HN90 or the bacterial agent as claimed in claim 2 in the preparation of biological organic fertilizer and / or water fertilizer.

4. Bacillus zanthoxylum as claimed in claim 1 Bacillus zanthoxyli Use of strain HN90 or the bacterial agent as claimed in claim 2 in promoting rice growth.

5. Bacillus zanthoxylum as claimed in claim 1 Bacillus zanthoxyli Use of strain HN90 or the bacterial agent as claimed in claim 2 in producing IAA, phosphate solubilization, phosphate dissolution and / or nitrogen fixation.

6. A method for preparing water-fertilizer using Bacillus zanthoxyli strain HN90 as claimed in claim 1 or the bacterial agent as claimed in claim 2, characterized in that: The method comprises: preparing Bacillus zanthoxylum HN90 into a suspension and then diluting it with sterile water to OD 600 =1.

7. Use the Bacillus zanthoxylum as claimed in claim 1 Bacillus zanthoxyli The method for preparing biological organic fertilizer by strain HN90 or the bacterial agent as claimed in claim 2, characterized in that: The method comprises the following steps: inoculating a bacterial liquid of Bacillus zanthoxylum HN90 into common organic fertilizer for fermentation to obtain biological organic fertilizer.

8. Use the Bacillus zanthoxylum as claimed in claim 1 Bacillus zanthoxyli The method for reducing fertilizer use in rice cultivation using strain HN90 or the bacterial agent as claimed in claim 2 is characterized in that: The method comprises the following steps: when planting rice, the amount of nitrogen fertilizer applied is reduced by 20% compared with the amount of nitrogen fertilizer applied in conventional fertilizers, and then 10 L / mu-20 L / mu of a compound solution of Bacillus zanthoxylum HN90 and GABA is applied in the field during the tillering period; the amount of nitrogen fertilizer applied in conventional fertilizers is 300 kg / ha.

9. The method according to claim 8, characterized in that The content of Bacillus zanthoxylum HN90 is 1×10 9 CFU / mL, and the GABA content is 1 g / L.

Images