Arsenic-resistant microbial bacterium with plant growth promoting function, complex microbial inoculant and preparation and application of arsenic-resistant microbial bacterium and complex microbial inoculant

Abstract

The invention discloses an arsenic-resistant microbial bacterium with a plant growth promoting function, a complex microbial inoculant and preparation and application of the arsenic-resistant microbial inoculant, the arsenic-resistant microbial inoculant comprises a cellulosimicrobium fibrosis bacterial strain Fh and a pseudomonas fluorescens bacterial strain Ff, the cellulosimicrobium fibrosis bacterial strain Fh is preserved in China Center for Type Culture Collection on June 26, 2024, and the pseudomonas fluorescens bacterial strain Ff is preserved in China Center for Type Culture Collection on June 26, 2024. The preservation address is Wuhan University, Wuhan, China, and the preservation number is CCTCC NO: M 20241367; the pseudomonas fluorescens strain Ff is preserved in the China Center for Type Culture Collection on June 26, 2024, the preservation address is Wuhan University, Wuhan, China, and the preservation number is CCTCC NO: M 20241366. The invention solves the problem of poor arsenic resistance of the existing growth-promoting microorganisms. After the complex microbial inoculant is applied, the dry weight of plants, the arsenic content of the overground part of ramie, and the effective arsenic content and the effective phosphorus content in soil are remarkably improved.

Description

Technical Field

[0001] The invention relates to an arsenic-resistant microorganism, and in particular to an arsenic-resistant microorganism with a plant growth-promoting function, a composite bacterial agent, and preparation and application thereof. Background Art

[0002] Industrial production and agricultural activities cause a large amount of arsenic to enter the soil and cause serious pollution. For the human body, food intake, respiratory tract and skin pores are the main ways for As to enter. As can induce damage to the structure of DNA chains, interfere with normal cell metabolism and inhibit the synthesis of some enzymes. When the human body ingests As exceeding a certain threshold, it will cause poisoning, ranging from vomiting, dizziness and general fatigue to gastrointestinal bleeding, nervous system weakness and liver damage in severe cases. When the human body is exposed to As for a long time, the risk of skin cancer, lung cancer, kidney cancer and bladder cancer will be greatly increased.

[0003] In recent years, bioremediation has become the main treatment method for arsenic-contaminated farmland soil due to its low investment and environmentally friendly advantages. Among them, phytoremediation technology with plant extraction as the core is considered to be the most effective remediation method. Phytoremediation often has the limitations of long remediation cycles and inhibited growth. In order to improve the effect of phytoremediation, existing technologies use other remediation methods to assist, such as electrokinetics and the addition of activators, but these methods usually cause certain damage to the functional structure of the soil. Therefore, the technology of enhancing phytoremediation through microorganisms is a green and friendly remediation technology with little soil disturbance. However, the existing growth-promoting microorganisms have poor arsenic resistance, and the resources of arsenic-resistant growth-promoting microorganisms are still in a relatively scarce stage. Summary of the invention

[0004] The purpose of the present invention is to provide an arsenic-resistant microorganism with plant growth-promoting function, a composite bacterial agent, and the preparation and application thereof, which solves the problem that the existing growth-promoting microorganisms have poor arsenic resistance. The composite bacterial agent has arsenic resistance, can survive in arsenic-contaminated soil and play a role in promoting plant growth, thereby improving the efficiency of plant restoration of arsenic-contaminated soil.

[0005] In order to achieve the above-mentioned object, the present invention provides an arsenic-resistant microorganism with plant growth-promoting function, the arsenic-resistant microorganism comprising Cellulosimicrobiun sp.Fh and Pseudomonas sp.Ff. The Cellulosimicrobiun sp.Fh was deposited in the China Center for Type Culture Collection on June 26, 2024, with a preservation address of Wuhan University, Wuhan, China, and the preservation numbers are CCTCC NO: M20241367; the Pseudomonas sp.Ff was deposited in the China Center for Type Culture Collection on June 26, 2024, with a preservation address of Wuhan University, Wuhan, China, and the preservation numbers are CCTCC NO: M 20241366.

[0006] The invention provides a composite bacterial agent, which is prepared by culturing the arsenic-resistant microbial bacteria with plant growth-promoting function.

[0007] The present invention provides a method for preparing the composite bacterial agent, the method comprising:

[0008] (1) The fibroblastic fibroblast strain Fh and the fluorescing Pseudomonas strain Ff as described in claim 1 are inoculated into a solid culture medium under sterile conditions, wherein the fibroblastic fibroblast strain Fh is cultured at 30° C. and the fluorescing Pseudomonas strain Ff is cultured at 25° C.;

[0009] (2) inoculating the cultured fibroblastic microbacterium strain Fh and the fluorescing Pseudomonas strain Ff into a basic liquid culture medium under sterile conditions, respectively, the fibroblastic microbacterium strain Fh is cultured at 30° C. and 150 rpm with shaking, and the fluorescing Pseudomonas strain Ff is cultured at 25° C. and 150 rpm with shaking;

[0010] (3) The fibrous fiber microbacterium strain Fh and the fluorescent Pseudomonas strain Ff after shaking culture are inoculated into the same basic liquid culture medium under sterile conditions, cultured at 28°C and 150 rpm, centrifuged at 4000 rpm, and the bacteria are collected. After washing, a bacterial suspension with OD600=1 is prepared with distilled water, i.e., the composite bacterial agent.

[0011] Preferably, in step (1), during inoculation, the volume of the fibroblastic fibroblast strain Fh is 2% of the volume of the solid culture medium; during inoculation, the volume of the fluorescing Pseudomonas strain Ff is 2% of the volume of the solid culture medium.

[0012] Preferably, in step (1), the components of the solid culture medium are tryptone, yeast extract powder, NaCl and agar, the mass ratio of tryptone, yeast extract powder, NaCl and agar is 2:1:2:3, and the culture time is 2 days.

[0013] Preferably, in step (2), the culturing time is 2 days; during inoculation, the volume of the fibroblastic fibroblast strain Fh after culture is 0.5% of the volume of the basic liquid culture medium; during inoculation, the volume of the fluorescing Pseudomonas strain Ff after culture is 0.5% of the volume of the basic liquid culture medium.

[0014] Preferably, in step (2), the components of the basic liquid culture medium are tryptone, yeast extract powder and NaCl, and the mass ratio of the tryptone, yeast extract powder and NaCl is 2:1:2.

[0015] Preferably, in step (3), the components of the basic liquid culture medium are peptone, yeast extract powder and NaCl, the mass ratio of the peptone, yeast extract powder and NaCl is 2:1:2, and the culturing time is 1 day; during inoculation, the volume of the fibrous fiber microbacterium strain Fh after shaking culture is 2% of the volume of the basic liquid culture medium; during inoculation, the volume of the fluorescing Pseudomonas strain Ff after shaking culture is 2% of the volume of the basic liquid culture medium.

[0016] The present invention provides an application of the arsenic-resistant microorganism with plant growth-promoting function or the composite bacterial agent in repairing arsenic pollution.

[0017] Preferably, the use comprises remediation of arsenic in soil or arsenic in contaminated water.

[0018] The arsenic-resistant microorganisms and composite bacterial agents with plant growth-promoting functions of the present invention, as well as their preparation and application, solve the problem of poor arsenic resistance of existing growth-promoting microorganisms, and have the following advantages:

[0019] 1. The present invention adopts trivalent and pentavalent arsenic selective culture medium, and provides specific guidance for microbial functions in the process of strain isolation and performance identification, thereby improving the efficiency and reliability of screening out target strains. Arsenic-resistant strains Fh and Ff with plant growth-promoting effects are obtained through growth-promoting function determination, and their actual effects are tested by combining them with the restoration plant ramie.

[0020] 2. The arsenic remediation plant selected in the present invention is the perennial plant ramie. Compared with the arsenic hyperaccumulator plant centipede grass commonly used in research, it can better adapt to the farmland environment. As a non-edible economic crop, it can ensure that the farmland has a certain economic output while carrying out plant remediation.

[0021] 3. Compared with the blank group, after applying the composite microbial agent of the present invention, the dry weight of the plant increased by 63.73%, the arsenic content in the aboveground part of ramie increased by 36.6%, and the effective arsenic content in the soil increased by 46.71%, thereby promoting the efficiency of arsenic migration from the soil to the roots of plants; at the same time, the arsenic removal rate of the soil applied with the composite microbial agent was significantly improved, reaching 2.07 times that of the soil without the application of the microbial agent; the effective phosphorus content of the soil increased by 41.92% compared with the control; at the same time, the alkaline nitrogen content of the soil was significantly higher than that of the soil applied with the microbial agent Ff, indicating that the nitrogen fixation function of the microorganism Fh can still play a stable role after the composite microbial agent is made and applied. Therefore, the composite microbial agent has arsenic resistance, can survive in arsenic-contaminated soil and play the ability to promote plant growth, thereby improving the efficiency of phytoremediation of arsenic-contaminated soil. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is the effective phosphorus content in the soil after the bacterial agent of the present invention is applied.

[0023] Figure 2 It is the alkaline nitrogen content of the soil after the bacterial agent of the present invention is applied.

[0024] Figure 3 This is a colony morphology diagram of the strain Fh of the present invention.

[0025] Figure 4 This is a colony morphology diagram of the strain Ff of the present invention.

[0026] Figure 5 It is a scanning electron microscope image of the strain Fh of the present invention.

[0027] Figure 6 It is a scanning electron microscope image of the strain Ff of the present invention.

[0028] Figure 7 This is the phylogenetic tree of the strain Fh of the present invention.

[0029] Figure 8 This is the phylogenetic tree of the strain Ff of the present invention.

[0030] Fig. 9 It is the ramie biomass after application of the bacterial agent of the present invention.

[0031] Fig.10 It is the arsenic content in the aerial part of ramie after application of the bacterial agent of the present invention.

[0032] Fig.11 It is the content of available arsenic in the soil after application of the bacterial agent of the present invention.

[0033] Fig.12 It is the soil pH after applying the bacterial agent of the present invention. DETAILED DESCRIPTION

[0034] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] The culture medium involved in the following examples is as follows:

[0036] Arsenic content: 1000 mg L -1 Selection medium: Each liter of culture medium contains 10 g of tryptone, 5 g of yeast extract powder, 10 g of NaCl, 15 g of agar, 4.164 g of sodium arsenate (or 1.734 g of sodium arsenite), and is adjusted to pH 7.0.

[0037] Inorganic phosphorus medium: Each liter of medium contains C6H 12 O610 g, Ca3(PO4)25g, MgCl25 g, MgSO4·7H2O0.25g, KCl 0.2g, (NH4)2SO40.1 g.

[0038] Basic solid culture medium: Each liter of culture medium contains 10g of tryptone, 5g of yeast extract powder, 10g of NaCl, and 15g of agar.

[0039] Basic liquid culture medium: Each liter of culture medium contains 10g of tryptone, 5g of yeast extract powder, and 10g of NaCl.

[0040] Example 1 Arsenic-resistant microorganisms and composite bacterial agents with plant growth-promoting functions

[0041] 1. Screening

[0042] 10 g of heavy metal-contaminated farmland soil sample from Liangshan Prefecture, Sichuan Province was placed in a 250 mL conical flask filled with 90 mL of sterile water, placed in a constant temperature incubator shaker at 30 °C and 150 rpm for 30 min, left to stand for 15 min, and the supernatant was diluted with sterile water to a concentration of 10 -1 , 10 -2 , 10 -3 and 10 -4 , draw 200 μL and apply it on As 5+ Content 1000mg L -1 After culturing for 2 to 3 days, the colonies grown on the culture medium were inoculated into As 3+ Content 1000mg L -1The microorganisms that can still grow normally were cultured in a selective medium, and the strains were continuously purified to a single strain. The strains were inoculated into an inorganic phosphorus medium at a 2% inoculum, and two arsenic-resistant strains with high phosphorus solubility were screened and named Cellulosimicrobiunsp.Fh and Pseudomonas sp.Ff. The colonies of the two strains were round, about 2 mm in diameter, and the surface was moist and smooth, of which Fh was yellow and Ff was light pink.

[0043] like Figure 3 As shown, the colony morphology of the strain Fh of the present invention. Figure 3 It can be seen that the diameter of Fh colonies is about 2 mm, and a single colony is round with neat edges, yellow in color, and a moist and smooth surface.

[0044] like Figure 4 As shown, the colony morphology of strain Ff of the present invention. Figure 4 It can be seen that the diameter of the Ff colony is about 2 mm, and a single colony is round with neat edges, light pink in color, and a moist and smooth surface.

[0045] like Figure 5 As shown, the electron microscope scanning image of the strain Fh of the present invention. Figure 5 It can be seen that the length of Fh is about 0.5-1.0μm, with a smooth surface and a bean shape.

[0046] like Figure 6 As shown, the electron microscope scanning image of the strain Ff of the present invention. Figure 6 It can be seen that the length of Ff is about 1.0-4.0 μm, the surface is slightly rough, and it is rod-shaped.

[0047] 2. Identification of bacterial species

[0048] The strain was inoculated into the basic liquid culture medium, and the bacterial liquid was used as a template, and PCR amplification was performed using universal primers 27F and 1492R (the nucleotide sequences of the universal primers 27F / 1492R are shown in SEQ ID NO.1 and SEQ ID NO.2). The reaction system was: 2 μL of strain DNA, 1 μL of each primer, 10 μL of 2×EasyTaq PCR SuperMix, and 11 μL of dd H2O; the reaction conditions were: initial 95°C pre-denaturation for 10 min, then 94°C denaturation for 30 s, 55°C annealing for 30 s, and 72°C extension for 90 s. This amplification step was repeated for 30 cycles, and the PCR reaction was terminated by extension at 72°C for 10 min. The PCR amplification product of the 16s rDNA gene of the strain was sequenced on the Illumina platform NovaSeq 6000 sequencer. The sequencing results were compared for sequence homology on the NCBI website.

[0049] like Figure 7 As shown, the phylogenetic tree of the strain Fh of the present invention;

[0050] like Figure 8 Shown is the phylogenetic tree of strain Ff of the present invention.

[0051] Depend on Figures 7-8 It can be seen that strain Fh is Cellulosimicrobium cellulans, belonging to the genus Cellulosimicrobium, with a homology of 99.43%, and Ff is Pseudomonasfluorescens, with a homology of 99.30%, belonging to the genus Pseudomonas.

[0052] Microbial strain Fh16 S The full-length rDNA sequence is shown in SEQ ID NO.3. S The full-length sequence of rDNA is shown in SEQ ID NO.4.

[0053] The microbial strains Fh and Ff were deposited in the China Center for Type Culture Collection on June 26, 2024, with the deposit address being Wuhan University, Wuhan, China, with the deposit numbers being CCTCC NO: M 20241367 and CCTCC NO: M 20241366, respectively.

[0054] 3. Preparation method of microbial composite agent

[0055] (1) Planar culture: The original strains of Microbacterium fibrosus Fh and Pseudomonas fluorescens Ff were inoculated into basic solid culture medium at a 2% inoculum under sterile conditions. Microbacterium fibrosus Fh was cultured at 30°C for 2 days, and Pseudomonas fluorescens Ff was cultured at 25°C for 2 days.

[0056] (2) Seed liquid culture: The bacterial strains cultured in step (1) were inoculated into the basic liquid culture medium at a 0.5% inoculum under sterile conditions. Fibromicrobacterium fibrosus Fh was cultured at 30°C with shaking at 150 rpm for 1 day, and Pseudomonas fluorescens Ff was cultured at 25°C with shaking at 150 rpm for 1 day.

[0057] (3) Mixed culture: Microbacterium fibrosus Fh and Pseudomonas fluorescens Ff were inoculated into the same bottle of basic liquid culture medium under sterile conditions at an inoculum volume ratio of 2% of the liquid culture medium. The culture was cultured at 150 rpm for 24 h at 28 °C and then centrifuged at 4000 rpm for 10 min. The cells were collected, washed three times, and then the OD was calculated using distilled water. 600= 1 bacterial suspension, that is, the composite bacterial agent. The inoculation volume is 2% of the volume of the basic liquid culture medium after shaking culture of the fibrous fiber microbacterium Fh or the fluorescent pseudomonas Ff.

[0058] 4. Preparation method of microbial agent Fh or Ff

[0059] This method is basically the same as the preparation method of the above-mentioned microbial composite agent, except that:

[0060] In step (3), the fibroblasts Fh and Pseudomonas fluorescens Ff were inoculated into different basal liquid media with the same content under sterile conditions at an inoculation rate of 2% of the volume of the liquid culture medium, cultured at 150 rpm for 24 h at 28° C., centrifuged at 4000 rpm for 10 min, and the cells were collected, washed three times, and then OD was obtained with distilled water. 600 =1, that is, microbial agent Fh or microbial agent Ff.

[0061] Experimental Example 1 Effect of microorganisms on ramie remediation of arsenic-contaminated soil

[0062] The components and contents of each component in arsenic-contaminated soil: pH 6.73, total carbon content 21.31 g / kg, total nitrogen content 1.04 g / kg, total phosphorus content 0.40 g / kg, total arsenic content 59.65 mg / kg, and effective arsenic content 0.30 mg / kg. The pot experiment was carried out in a planting greenhouse. Each flower pot was filled with 0.5 kg of arsenic-contaminated soil. Ramie seedlings were carefully cleaned of the root soil and transplanted into the pot. 200 mL of distilled water was poured to fix it. The survival of ramie was observed after 10 days. After the ramie seedlings survived the transplant, 100 mL of composite bacterial agent was applied as the composite bacterial agent group, 100 mL of microbial agent Fh was applied as the bacterial agent Fh group, 100 mL of microbial agent Ff was applied as the bacterial agent Ff group, and 100 mL of distilled water was used as the blank group. Six replicates were set up for each group. After 40 days of cultivation, the ramie plants were harvested and ramie and soil samples were collected.

[0063] Determination indicators of ramie samples: ramie dry weight, arsenic content in the above-ground and underground parts of ramie.

[0064] Soil sample measurement indicators: available arsenic content, pH value, available phosphorus content, and alkaline nitrogen content.

[0065] like Fig. 9 As shown, the ramie biomass after the application of the microbial agent of the present invention, wherein the blank is the blank group, the microbial agent Fh is the group applied with the microbial agent Fh, the microbial agent Ff is the group applied with the microbial agent Ff, and the composite microbial agent is the group applied with the composite microbial agent. Fig. 9It can be seen that the application of microbial agents effectively promoted the growth of ramie. Compared with the blank group, the biomass of ramie in the experimental group with the application of microbial agents increased significantly, among which the application of composite microbial agents had the most significant effect, with the dry weight of the plants increasing by 63.73% compared with the control, and the growth-promoting effect was obvious.

[0066] like Fig.10 As shown, the arsenic content in the aboveground part of ramie after the application of the bacterial agent of the present invention, wherein the blank is the blank group, the bacterial agent Fh is the group applied with the bacterial agent Fh, the bacterial agent Ff is the group applied with the bacterial agent Ff, and the composite bacterial agent is the group applied with the composite bacterial agent. Fig.10 It can be seen that compared with the blank group, the application of bacterial agents also played a certain role in promoting arsenic absorption by ramie. After the application of the composite bacterial agent, the arsenic content in the aboveground part of ramie increased by 36.6%.

[0067] like Fig.11 As shown, the effective arsenic content in the soil after the application of the bacterial agent of the present invention, wherein the blank is the blank group, the bacterial agent Fh is the group applied with the bacterial agent Fh, the bacterial agent Ff is the group applied with the bacterial agent Ff, and the composite bacterial agent is the group applied with the composite bacterial agent. Fig.11 It can be seen that compared with the blank group, the content of available arsenic in the soil was significantly increased after the application of the composite bacterial agent, which increased by 46.71%. This increase is one of the main factors that promote the efficiency of arsenic absorption by ramie.

[0068] like Fig.12 As shown, the soil pH after the bacterial agent of the present invention was applied, wherein the blank is the blank group, the bacterial agent Fh is the group applied with the bacterial agent Fh, the bacterial agent Ff is the group applied with the bacterial agent Ff, and the composite bacterial agent is the group applied with the composite bacterial agent. Fig.12 It can be seen that the application of microbial agents has the effect of increasing soil pH, which has a promoting effect on soil arsenic activation.

[0069] like Figure 1 As shown, the soil available phosphorus content after the application of the bacterial agent of the present invention, wherein the blank is the blank group, the bacterial agent Fh is the group applied with the bacterial agent Fh, the bacterial agent Ff is the group applied with the bacterial agent Ff, and the composite bacterial agent is the group applied with the composite bacterial agent. Figure 1 It can be seen that the application of microbial agents significantly increased the available phosphorus content in the soil, and the composite agent showed a more significant improvement effect, with the available phosphorus content in the soil increased by 41.92% compared with the control.

[0070] like Figure 2 As shown, the soil alkaline nitrogen content after the application of the bacterial agent of the present invention, wherein the blank is the blank group, the bacterial agent Fh is the group applied with the bacterial agent Fh, the bacterial agent Ff is the group applied with the bacterial agent Ff, and the composite bacterial agent is the group applied with the composite bacterial agent. Figure 2It can be seen that the alkaline nitrogen content of the soil after the composite microbial agent was applied was significantly higher than that of the soil applied with the microbial agent Ff, indicating that the nitrogen fixation function of the microorganism Fh can still play a stable role after the composite microbial agent is applied. Compared with the large-scale use of nitrogen and phosphorus fertilizers in the existing plant restoration process, the microbial composite microbial agent of the present invention has an improvement effect on soil nutrients, can reduce the use of fertilizers to a certain extent, and try to avoid the use of common phosphate fertilizers during restoration, which leads to the input of arsenic-containing phosphate rock.

[0071] Although the content of the present invention has been described in detail through the above preferred embodiments, it should be appreciated that the above description should not be considered as a limitation of the present invention. After reading the above content, it will be apparent to those skilled in the art that various modifications and substitutions of the present invention will occur. Therefore, the protection scope of the present invention should be limited by the appended claims.

Claims

1. An arsenic-resistant microorganism with plant growth-promoting function, characterized in that: The arsenic-resistant microorganisms include Cellulosimicrobiun sp. Fh and Pseudomonas sp. Ff. The Cellulosimicrobiun sp. Fh was deposited in the China Center for Type Culture Collection on June 26, 2024, with a preservation address of Wuhan University, Wuhan, China, and the preservation numbers are CCTCC NO: M 20241367; the Pseudomonas sp. Ff was deposited in the China Center for Type Culture Collection on June 26, 2024, with a preservation address of Wuhan University, Wuhan, China, and the preservation numbers are CCTCC NO: M 20241366.

2. A composite bacterial agent, characterized in that: The composite bacterial agent is prepared by culturing the arsenic-resistant microbial bacteria with plant growth-promoting function as claimed in claim 1.

3. A method for preparing the composite bacterial agent according to claim 2, characterized in that: The method includes: (1) The fibroblastic fibroblast strain Fh and the fluorescing Pseudomonas strain Ff as described in claim 1 are inoculated into a solid culture medium under sterile conditions, wherein the fibroblastic fibroblast strain Fh is cultured at 30° C. and the fluorescing Pseudomonas strain Ff is cultured at 25° C.; (2) inoculating the cultured fibroblastic microbacterium strain Fh and the fluorescing Pseudomonas strain Ff into a basic liquid culture medium under sterile conditions, respectively, the fibroblastic microbacterium strain Fh is cultured at 30° C. and 150 rpm with shaking, and the fluorescing Pseudomonas strain Ff is cultured at 25° C. and 150 rpm with shaking; (3) The fibrous fiber microbacterium strain Fh and the fluorescent Pseudomonas strain Ff after shaking culture are inoculated into the same basic liquid culture medium under sterile conditions, cultured at 28°C and 150 rpm, centrifuged at 4000 rpm, and the bacteria are collected. After washing, a bacterial suspension with OD600=1 is prepared with distilled water, i.e., the composite bacterial agent.

4. The preparation method according to claim 3, characterized in that: In step (1), during inoculation, the volume of the fibroblastic fibrosis microbacterium strain Fh is 2% of the volume of the solid culture medium; during inoculation, the volume of the fluorescing Pseudomonas strain Ff is 2% of the volume of the solid culture medium.

5. The preparation method according to claim 3, characterized in that: In step (1), the components of the solid culture medium are tryptone, yeast extract powder, NaCl and agar, the mass ratio of tryptone, yeast extract powder, NaCl and agar is 2:1:2:3, and the culture time is 2 days.

6. The preparation method according to claim 3, characterized in that: In step (2), the culture time is 2 days; during inoculation, the volume of the fibroblastic fibroblast strain Fh after culture is 0.5% of the volume of the basic liquid culture medium; during inoculation, the volume of the fluorescing Pseudomonas strain Ff after culture is 0.5% of the volume of the basic liquid culture medium.

7. The preparation method according to claim 3, characterized in that: In step (2), the components of the basic liquid culture medium are tryptone, yeast extract powder and NaCl, and the mass ratio of the tryptone, yeast extract powder and NaCl is 2:1:

2.

8. The preparation method according to claim 3, characterized in that: In step (3), the components of the basic liquid culture medium are peptone, yeast extract powder and NaCl, the mass ratio of the peptone, yeast extract powder and NaCl is 2:1:2, and the culture time is 1 day; during inoculation, the volume of the fibrous fiber microbacterium strain Fh after shaking culture is 2% of the volume of the basic liquid culture medium; during inoculation, the volume of the fluorescing Pseudomonas strain Ff after shaking culture is 2% of the volume of the basic liquid culture medium.

9. Use of the arsenic-resistant microorganism with plant growth-promoting function as claimed in claim 1 or the composite bacterial agent as claimed in claim 2 in remediating arsenic pollution.

10. The use according to claim 9, characterized in that: The applications include remediation of arsenic in soil or arsenic in contaminated water.

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