Phosphorus-solubilizing bacteria, phosphate-solubilizing bacteria agent and application thereof

Abstract

The invention relates to the field of environment and agricultural biotechnology, in particular to a phosphate-solubilizing bacterium, a phosphate-solubilizing microbial agent and application of the phosphate-solubilizing bacterium, the classification name of a phosphate-solubilizing bacterium strain HS-6 is Enterobacter hormaechei, the strain HS-6 is preserved in the China General Microbiological Culture Collection Center on October 19, 2023, and the preservation number is CGMCC NO.28680. The invention further discloses a preparation method of the phosphate-solubilizing bacterium, the phosphate-solubilizing microbial agent and application of the phosphate-solubilizing bacterium and the phosphate-solubilizing microbial agent. The bacterial colony of the strain HS-6 has the characteristics of irregular circle, semitransparent property, bulge, irregular edge, milk white, short rod shape and gram negative bacterium. Besides, after the strain HS-6 is subjected to culture condition optimization, the phosphorus solubilizing amount can reach 821.57 mg / L. Therefore, the phosphorus solubilizing bacterium has the advantage of high phosphorus solubilizing capacity, can be used for preparing a phosphorus solubilizing bacterium agent, has a good application prospect, and particularly has a good application prospect in the aspects of dissolving phosphate ores and developing phosphorus solubilizing bacterium fertilizers.

Description

Technical Field

[0001] The invention relates to the fields of environment and agricultural biotechnology, and in particular to phosphate-dissolving bacteria, a phosphate-dissolving bacterial agent and applications thereof. Background Art

[0002] Phosphorus is an important biological element that plays a vital role in the biosynthesis of various compounds such as phytochemicals, nucleic acids and phospholipids. It is closely related to nitrogen and carbon metabolism as well as photosynthesis, thus having a positive impact on crop growth and development. Insufficient phosphorus in crops will hinder the synthesis of nucleic acids, phospholipids and phosphate nucleotides, thereby affecting cell division, energy conversion and cellular respiration, and ultimately leading to a decrease in the photosynthetic rate. The average concentration of inorganic phosphorus available to plants in the soil is usually in the range of 1μmol / L to 10μmol / L, which is far lower than the optimal phosphorus concentration required for crop growth. In addition, the soil contains a certain proportion of organic phosphorus (30% to 80% of the total phosphorus), but most of it is unavailable to crops.

[0003] Soil microorganisms control key bio-geo-chemical processes in agricultural ecosystems and play a vital role in nutrient cycling and energy flow. Phosphate-solubilizing bacteria is a general term for microorganisms that have the ability to decompose phosphorus-containing compounds and promote the effectiveness of phosphorus, which can improve the utilization efficiency of phosphate fertilizers and phosphate rock. Phosphate-solubilizing bacteria are generally considered to be a high-quality alternative to chemical phosphate fertilizers. While promoting plant absorption of phosphorus, phosphate-solubilizing bacteria not only do not harm the agricultural ecological environment, but also have the potential to repair the soil environment polluted by traditional commercial fertilizers. There are many types of phosphate-solubilizing microorganisms, which can be divided into phosphate solubilizing bacteria (PSB), phosphate solubilizing fungi (PSF) and phosphate solubilizing actinomyces (PSA). Among them, phosphate-solubilizing bacteria are the most numerous, accounting for about 1% to 50% of the total number of phosphate-solubilizing microorganisms. Compared with phosphate-solubilizing bacteria, the number of phosphate-solubilizing fungi is relatively small, accounting for only 0.1% to 0.5%.

[0004] According to existing literature reports, phosphate-dissolving bacteria are widely distributed and strains with strong abilities are not concentrated. Different types of bacterial resources may contain strains with high phosphate-dissolving activity. From this perspective, the screening of wild strains with phosphate-dissolving ability is still of great significance. In general, the wild strains used for industrial production and development are extremely single. Except for the phosphate-dissolving Bacillus megaterium, which has certain development and application, the research on other strains is far from systematic and in-depth. In terms of the screening of phosphate-dissolving bacteria, most of them use a single inorganic phosphorus culture medium or a single organic phosphorus culture medium for screening, which limits the phosphate-dissolving ability of the strain. In addition, the strain resource library of phosphate-dissolving bacteria needs to be further enriched, and phosphate-dissolving bacterial fertilizers need to be further developed. Summary of the invention

[0005] In order to overcome the deficiencies of the prior art, the first object of the present invention is to provide a phosphate-dissolving bacterium. The phosphate-dissolving bacterium further enriches the genetic resources of wild phosphate-dissolving bacteria, expands the reserve library of phosphate-dissolving bacteria whole genome breeding, obtains live pure cultures of wild strains with strong phosphate-dissolving ability, further enriches the strain resource library of phosphate-dissolving bacteria, and is of great significance for the development of phosphate-dissolving bacterial fertilizers.

[0006] In order to overcome the shortcomings of the prior art, the second object of the present invention is to provide a phosphate-dissolving bacteria agent.

[0007] The third object of the present invention is to provide an application of phosphate-dissolving bacteria.

[0008] In order to achieve the first object of the above invention, the technical solution adopted by the present invention is as follows:

[0009] The present invention provides a phosphate-dissolving bacterium, and the classification name of the phosphate-dissolving bacterium strain HS-6 is Enterobacter hormaechei. The strain HS-6 was deposited in the General Microbiology Center of China National Microbiological Culture Collection Committee on October 19, 2023, with a deposit number of CGMCC NO.: 28680.

[0010] The colony characteristics of the strain HS-6 are: irregular circle, translucent, convex, irregular edge, milky white, short rod-shaped, Gram-negative bacteria. Its growth characteristics are: the growth curve is basically "S" type. In addition, the phosphate dissolving amount of the strain HS-6 can reach 821.57 mg / L after the culture conditions are optimized. Therefore, a phosphate dissolving bacterium of the present invention has the advantage of strong phosphate dissolving ability and has a good application prospect, especially in the application prospect of dissolving phosphate ore and developing phosphate dissolving bacterial fertilizer.

[0011] Further, the 16S rDNA gene sequence of the phosphate-dissolving bacteria is shown in SEQ ID No. 1. Based on the gene sequencing results and the physiological and biochemical experimental results, it is determined that the phosphate-dissolving bacteria belong to Enterobacter hormaechei. The colony characteristics of the phosphate-dissolving bacteria are irregularly round, translucent, convex, irregular edges, milky white, short rod-shaped, and Gram-negative bacteria.

[0012] In order to achieve the second purpose of the above invention, the technical solution adopted by the present invention is as follows:

[0013] The present invention provides a phosphate-dissolving bacteria agent, which contains the above-mentioned phosphate-dissolving bacteria. The phosphate-dissolving bacteria strain HS-6 is prepared into the phosphate-dissolving bacteria agent, so as to better utilize the phosphate-dissolving ability of the phosphate-dissolving bacteria strain HS-6.

[0014] In order to achieve the third purpose of the above invention, the technical solution adopted by the present invention is as follows:

[0015] The present invention provides the use of the above-mentioned phosphate-dissolving bacteria agent in dissolving phosphate ore.

[0016] The present invention provides the use of the above-mentioned phosphate-dissolving bacteria agent as an agricultural microbial agent fertilizer.

[0017] The present invention provides application of the above-mentioned phosphate-dissolving bacteria agent in environmental restoration.

[0018] The present invention provides the use of the above-mentioned phosphate-dissolving bacteria in dissolving phosphate ore.

[0019] The present invention provides the use of the above-mentioned phosphate-dissolving bacteria as agricultural microbial fertilizer.

[0020] The present invention provides the use of the above-mentioned phosphate-dissolving bacteria in environmental restoration.

[0021] Among them, since the phosphate-dissolving bacteria strain HS-6 of the present invention has a strong phosphate-dissolving ability, the above-mentioned phosphate-dissolving bacteria agent or phosphate-dissolving bacteria are applied to dissolving phosphate ore, the above-mentioned phosphate-dissolving bacteria agent or phosphate-dissolving bacteria are applied to the preparation of agricultural microbial fertilizers, and the above-mentioned phosphate-dissolving bacteria agent or phosphate-dissolving bacteria are applied to environmental remediation, all of which have good application prospects.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] (1) A phosphate-dissolving bacterium of the present invention is determined to belong to Enterobacter hormaechei through morphological observation, 16SrDNA homologous sequence and phylogenetic tree analysis of the phosphate-dissolving bacterium strain HS-6. The strain HS-6 further enriches the genetic resources of wild phosphate-dissolving bacteria, expands the reserve library of phosphate-dissolving bacteria whole genome breeding, obtains live pure cultures of wild strains with strong phosphate-dissolving ability, further enriches the strain resource library of phosphate-dissolving bacteria, and is of great significance for the development of phosphate-dissolving bacterial fertilizers.

[0024] (2) Application of a phosphate-dissolving bacterium of the present invention. Since the phosphate-dissolving bacterium has a strong phosphate-dissolving ability, the phosphate-dissolving amount of strain HS-6 can reach 821.57 mg / L after the culture conditions are optimized. The phosphate-dissolving bacterium also plays an important role in improving the ability of plants to cope with biological and non-biological stresses such as drought, salinity, heavy metals and pests and diseases. The phosphate-dissolving bacterium or the phosphate-dissolving agent containing the phosphate-dissolving bacterium is used to dissolve phosphate ore, develop phosphate-dissolving fertilizer or environmental remediation, which has good application prospects and important significance. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0026] Figure 1 It is the characteristic diagram of the phosphate solubilization circle of the strain HS-6 of the present invention.

[0027] Figure 2 It is a morphological diagram of the strain HS-6 of the present invention subjected to Gram staining.

[0028] Figure 3 It is a scanning electron micrograph of the strain HS-6 of the present invention.

[0029] Figure 4 It is a phylogenetic tree of strain HS-6 of the present invention.

[0030] Figure 5 It is a growth curve diagram of the strain HS-6 of the present invention.

[0031] Figure 6 This is a graph showing the results of detecting the amount of phosphorus solubilization of the strain HS-6 of the present invention when cultured at different temperatures.

[0032] Figure 7 This is a graph showing the results of detecting the amount of phosphorus solubilization of the strain HS-6 of the present invention when cultured at different initial pH values.

[0033] Figure 8 This is a graph showing the results of detecting the amount of phosphorus solubilization of the strain HS-6 of the present invention when cultured under different carbon sources.

[0034] Fig. 9 This is a graph showing the results of detecting the amount of phosphorus solubilization of the strain HS-6 of the present invention when cultured under different nitrogen sources. DETAILED DESCRIPTION

[0035] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0036] The terms used in the embodiments of the present invention are only for the purpose of describing specific embodiments, and are not intended to limit the present invention. In the present invention, the singular forms "a", "said" and "the" used in the embodiments and the appended claims are also intended to include plural forms, unless the context clearly indicates other meanings.

[0037] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0038] The culture medium formula described in the following examples is as follows:

[0039] PVK medium (g / L): NaCl 0.2g / L, Ca 3 (PO 4 ) 2 5g / L, MgSO 4 0.1g / L, (NH 4 ) 2 SO 4 0.5g / L, glucose 15g / L, pH 6.8~7.0.

[0040] Solid culture medium of PVK: Add 18g to 20g of agar powder to every 1L of PVK culture medium.

[0041] LB medium (g / L): peptone 10 g / L, yeast extract 5 g / L, NaCl 10 g / L, pH 7.0-7.2.

[0042] LB solid culture medium: Add 18g to 20g of agar powder per 1L of LB culture medium.

[0043] NBRIP medium (g / L): glucose 10 g / L, (NH 4 ) 2 SO 4 0.5g / L, NaCl 0.3g / L, MgSO 4 7H2 O0.3g / L, FeSO 4 7H 2 O 0.03g / L, MnSO 4 ·2H 2 O 0.03g / L, CaCO 3 5g / L, KCl 0.3g / L, lecithin 0.2g / L, pH 7.2~7.4.

[0044] NBRIP solid culture medium: Add 18g to 20g of agar powder to every 1L of NBRIP culture medium.

[0045] Among them, PVK medium was used as the separation medium, LB medium was used as the preservation medium, and NBRIP medium was used as the phosphate solubilization ability determination medium.

[0046] Example 1 Isolation and identification of strains

[0047] 1. Sample collection

[0048] Sample: Soil sample from Mingli Farm, Tianjin University of Technology (39°3′38″N, 117°7′48″E).

[0049] 2. Isolation and screening of strains

[0050] The separation and screening method adopts the enrichment culture method, which is as follows:

[0051] According to the soil bacteria isolation procedure, the collected soil samples were enriched and cultured at 30°C and 180 r / min in a shaking incubator for 30 min; then, under sterile operation conditions, the soil suspension after enrichment culture in the shaking incubator was gradiently diluted in sequence to obtain 10 -2 , 10 -3 , 10 -4 , 10 -5 and 10 -6 Soil suspension of equal dilution gradient. 100 μL of bacterial solution of different dilution concentrations was applied to PVK medium, with 3 parallels for each gradient, and inverted cultured at 30°C for 3 to 7 days; then single colonies with good growth, typical characteristics, and large and significant phosphate solubility zone were picked with an inoculation loop, and continued to be streaked and purified on solid separation medium (i.e., solid medium of PVK) to obtain pure culture of target strains and preserve them.

[0052] The above method was used to successfully isolate a phosphate-dissolving bacterial strain from environmental samples, which was numbered HS-6. The strain has a strong phosphate-dissolving ability, and the phosphate-dissolving capacity of strain HS-6 can reach 821.57 mg / L after the culture conditions were optimized.

[0053] 2. Strain identification

[0054] The isolated strain HS-6 with obvious phosphate solubility zone was Figure 1 The target strain was identified as shown in the figure. The phosphorus solubilizing ability of the strain was determined by the molybdenum antimony colorimetric method. After 3 days of culture, the phosphorus solubilizing capacity of the strain HS-6 in the culture medium supernatant reached 382 mg / L, indicating that the strain HS-6 has a strong phosphorus solubilizing ability.

[0055] Among them, in the process of determining the phosphorus solubilizing ability of the strain by the molybdenum antimony colorimetric method, the strain with the strongest phosphorus solubilizing ability was selected. After the purified phosphorus solubilizing bacteria were inoculated into the LB liquid culture medium for culture, the bacterial solution was inoculated into the NBRIP culture medium at a 1% (volume percentage) inoculation amount, and no inoculation of bacterial solution was set as a control, and 3 groups of parallel experiments were set. The culture was cultured at 30°C and 180r / min for 7 days, and samples were taken every 12h, and then centrifuged at 4°C and 2500r / min for 10min, and the supernatant was taken to determine the effective phosphorus content by the molybdenum antimony colorimetric method.

[0056] Among them, the colony characteristics of strain HS-6 are irregular round, translucent, convex, irregular edges, milky white, short rod-shaped, and Gram-negative bacteria. Figure 2 and Figure 3 As shown, Figure 2 The morphological diagram of strain HS-6 after Gram staining is shown in Figure 2. Figure 3 shown.

[0057] 16S rDNA molecular biological identification:

[0058] Among them, the 16SrDNA PCR amplification primers are:

[0059] Forward primer 27F: 5′-CAGAGTTTGATCCTGGCT-3′

[0060] Reverse primer 1540R: 5′-AGGAGGTGATCCAGCCGCA-3′

[0061] The target strain 16SrDNA was amplified by PCR, and the PCR products were screened by electrophoresis, gel recovery, plasmid vector ligation, transgenic, and culture. Finally, the positive clones were sequenced to obtain the 16S rDNA gene sequence of the HS-6 strain, which was 1476 in length. Please refer to the sequence table for its DNA sequence.

[0062] Among them, the 16S rDNA gene sequence of strain HS-6 was searched for homology through the blast online analysis function of NCBI, and the phylogenetic tree was constructed using MEGA software.

[0063] Phylogenetic tree of 16S rDNA Figure 4 As shown, the result shows that the strain HS-6 obtained by separation and purification of the present invention belongs to Enterobacter hormaechei. There is no literature report on the ability of Enterobacter hormaechei with respect to the colony morphology, so the screening of strain HS-6 enriches the genetic resources of wild phosphate-dissolving bacteria and expands the reserve library of phosphate-dissolving bacteria full genome breeding. The gene sequence of strain HS-6 has been submitted to the Gen Bank database, and the accession number is PQ826318.

[0064] In addition, the strain HS-6 was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on October 19, 2023, with the deposit number CGMCC NO.: 28680, and the deposit address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0065] Example 2 Identification of growth characteristics of strain HS-6

[0066] The HS-6 strain of phosphate-solubilizing bacteria was inoculated into 100 mL of LB liquid medium and cultured at 30°C and 180 rpm. The OD was measured every 2 h. 600 Optical density value, with culture time as the horizontal axis, OD 600 The optical density value is the ordinate, and the growth curve is drawn. The growth curve of HS-6 strain is shown in Figure 5 shown.

[0067] Depend on Figure 5 It can be seen that after a delay period of 0h to 6h, the HS-6 strain entered the rapid growth logarithmic phase at 6h to 20h. 600 When the value is between 0.6 and 0.8, it indicates that the HS-6 strain is in a vigorous logarithmic phase. The growth curve is basically "S" shaped.

[0068] Example 3 Optimization of culture conditions and determination of phosphate solubilization capacity of strain HS-6

[0069] Using NBRIP liquid medium as the basic medium, strain HS-6 was inoculated into the medium at a volume of 1% (volume percentage of the medium), and no inoculation was used as a blank control. The medium was cultured in a shaking incubator at 25°C, 28°C, and 30°C at 180 r / min for 3 days, and three parallel cultures were set. The supernatants were then centrifuged at 4°C and 2500 r / min for 10 min, and the effective phosphorus content was determined by the molybdenum antimony colorimetric method. The test results are shown in the following table. Figure 6 As shown. Figure 6 It can be seen that the single factor test results found that the most suitable growth temperature for the strain HS-6 of the present invention is 28°C.

[0070] Using NBRIP liquid medium as the basic medium, strain HS-6 was inoculated into the medium at a rate of 1% (volume percentage of the medium), and no inoculation was used as the blank control. The initial pH was set to 3, 5, 7, 8, and 9, respectively, and cultured at 28°C and 180 r / min in a shaking incubator for 3 days. Three parallels were set, and then centrifuged at 4°C and 2500 r / min for 10 min. The supernatant was taken and the effective phosphorus content was determined by the molybdenum antimony colorimetric method. The test results are shown in the figure. Figure 7 As shown. Figure 7 It can be seen that the single factor test results show that the most suitable initial pH value for the strain HS-6 of the present invention is 8.

[0071] Using NBRIP liquid medium as the basic medium, the strain HS-6 was inoculated with 1% (volume percentage of the medium) of bacterial solution in the medium, and no bacterial solution was used as the blank control. The carbon sources were selected as glucose, sucrose, fructose and soluble starch, respectively. The culture was carried out in a shaking incubator at 28°C and 180 r / min for 3 days, and three parallel cultures were set. Then, the culture was centrifuged at 4°C and 2500 r / min for 10 min, and the supernatant was taken to determine the effective phosphorus content by molybdenum antimony colorimetric method. The test results are shown in the figure. Figure 8 As shown. Figure 8 It can be seen that the utilization rate of different carbon sources by strain HS-6 is ranked in the order of glucose > sucrose > fructose > soluble starch.

[0072] Using NBRIP liquid medium as the basic medium, the strain HS-6 was inoculated with 1% (volume percentage of the medium) of bacterial solution in the medium, and no bacterial solution was used as the blank control. The nitrogen sources were selected from ammonium sulfate, ammonium chloride, potassium nitrate, peptone and urea, respectively. The culture was carried out at 28°C and 180r / min in a shaking incubator for 3 days, and three parallel cultures were set. Then, the culture was centrifuged at 4°C and 2500r / min for 10min, and the supernatant was taken to determine the effective phosphorus content by molybdenum antimony colorimetric method. The test results are shown in the figure. Fig. 9 As shown. Fig. 9 It can be seen that the utilization rate of different nitrogen sources by strain HS-6 is ranked as ammonium sulfate > ammonium chloride > urea > potassium nitrate > peptone.

[0073] In addition, based on the results of the above single factor test, an orthogonal test was conducted on the culture conditions of strain HS-6. The conditions of the orthogonal test and the corresponding phosphorus solubilization amount are shown in Table 1.

[0074] Table 1 Orthogonal test table of culture conditions of strain HS-6

[0075]

[0076]

[0077] The results in Table 1 combined with further experimental verification determined that the order of influence on the phosphorus solubilization ability of Enterobacter huoxigensis HS-6 is A>D>C>B, that is, temperature>ammonium sulfate concentration>glucose concentration>initial pH. The optimal levels of factors A, B, C, and D are A3, B3, C3, and D3, respectively, and the optimal combination is A3B3C3D3, that is, the culture temperature is 28℃, the initial pH is 8, the glucose concentration is 10g / L, and the ammonium sulfate concentration is 0.15g / L. Factors A and B in the optimal combination are consistent with the results of the single factor experiment, that is, the optimal culture temperature is 28℃ and the optimal initial pH is 8. Since the orthogonal experiment does not involve the optimal combination of Enterobacter huoxigensis, in order to verify the actuality of the optimal combination obtained by the orthogonal experiment theory, a supplementary experiment was carried out, with no inoculation as the blank control, and three parallel samples were set. The results show that when Enterobacter huoxigensis HS-6 was cultured under the optimal combination conditions, the phosphorus solubility reached a peak of 821.57 mg / L, which was the highest in all experimental groups, consistent with the theory.

[0078] Example 4

[0079] A phosphate-dissolving bacterial agent contains the phosphate-dissolving bacteria of the present invention. The phosphate-dissolving bacterial strain HS-6 is prepared into the phosphate-dissolving bacterial agent, so that the phosphate-dissolving ability of the phosphate-dissolving bacterial strain HS-6 can be better utilized.

[0080] Example 5

[0081] An application of phosphate-dissolving bacteria, wherein the phosphate-dissolving bacteria of the present invention is applied to dissolving phosphate ore, or the phosphate-dissolving bacteria of the present invention is applied to preparing agricultural microbial fertilizer, or the phosphate-dissolving bacteria of the present invention is applied to environmental remediation. Since the phosphate-dissolving bacteria strain HS-6 of the present invention has a strong phosphate-dissolving ability, the phosphate-dissolving bacteria of the present invention is applied to dissolving phosphate ore, preparing agricultural microbial fertilizer, or environmental remediation, all of which have good application prospects.

[0082] Example 6

[0083] An application of a phosphate-dissolving bacteria agent, wherein the phosphate-dissolving bacteria agent of the present invention is applied to dissolving phosphate ore, or the phosphate-dissolving bacteria agent of the present invention is applied to preparing agricultural microbial fertilizer, or the phosphate-dissolving bacteria agent of the present invention is applied to environmental remediation. Since the strain HS-6 in the phosphate-dissolving bacteria agent of the present invention has a strong phosphate-dissolving ability, the phosphate-dissolving bacteria agent of the present invention is applied to dissolving phosphate ore, preparing agricultural microbial fertilizer, or environmental remediation, all of which have good application prospects.

[0084] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

[0085]

Claims

1. A phosphate-dissolving bacterium, characterized in that: The phosphate-dissolving bacterial strain HS-6 is classified and named Enterobacter hormaechei. The strain HS-6 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on October 19, 2023, with the deposit number CGMCC NO.: 28680.

2. A phosphate-dissolving bacterium according to claim 1, characterized in that: The 16SrDNA gene sequence of the phosphate-dissolving bacteria is shown in SEQ ID No.

1.

3. A phosphate-dissolving bacteria agent, characterized in that: Containing the phosphate-dissolving bacteria according to claim 1 or 2.

4. Use of the phosphate-dissolving bacteria agent according to claim 3 in dissolving phosphate ore.

5. Use of the phosphate-dissolving bacteria agent according to claim 3 as an agricultural microbial fertilizer.

6. Use of the phosphate-dissolving bacteria agent according to claim 3 in environmental remediation.

7. Use of the phosphate-dissolving bacteria according to claim 1 or 2 in dissolving phosphate ore.

8. Use of the phosphate-dissolving bacteria according to claim 1 or 2 as agricultural microbial fertilizer.

9. Use of the phosphate-dissolving bacteria described in claim 1 or 2 in environmental remediation.

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