Microbial agent containing alcaligenes of impatiens balsamina and arthrobacter nicotianae and heavy metal passivation method

Abstract

The invention relates to a microbial agent containing impatiens balsamina alcaligenes and arthrobacter nicotianae and a heavy metal passivation method, the microbial agent comprises a main microbial agent and an auxiliary microbial agent, and the main microbial agent comprises the impatiens balsamina alcaligenes; the auxiliary microbial agent comprises arthrobacter nicotianae; the impatiens balsamina alcaligenes is classified and named as impatiens balsamina alcaligenes, and the preservation number of the impatiens balsamina alcaligenes is CGMCC (China General Microbiological Culture Collection Center) No.14474; the arthrobacter nicotianae is classified and named as Arthrobacter nicotinovorans, and the preservation number of the arthrobacter nicotinovorans is CGMCC (China General Microbiological Culture Collection Center) No.33646. The method disclosed by the invention has a relatively good passivation effect on heavy metals, and can adjust the pH value of a water and soil environment and improve the content of urease-producing bacteria in soil.

Description

Technical Field

[0001] This application belongs to the field of agricultural biotechnology. Specifically, it relates to a microbial inoculant including Alcaligenes balsami and Arthrobacter nicotianae, and a method for heavy metal passivation. Background Art

[0002] In recent decades, anthropogenic activities such as highly intensive agricultural production, industrial and mining activities, such as the unreasonable use of pesticides and fertilizers, and the large-scale discharge of "three wastes" from heavy metal-related enterprises, have led to an increasingly serious problem of heavy metal pollution in farmland soil. Soil heavy metal pollution is closely related to human activities. Soil heavy metals mainly include Cd, Cu, Zn, Hg, Pb, etc. Crops can absorb and accumulate heavy metals from farmland soil through their roots, and pose a potential threat to human health through the food chain. Heavy metal pollution in farmland soil can directly endanger food security and human health. Remediation of heavy metal-contaminated soil is a worldwide problem, and the "repair while producing" of moderately and lightly heavy metal-contaminated cultivated land is an important research direction and practical need for food production and food safety. In-situ passivation repair with high efficiency and environmental protection has become the preferred solution. Heavy metal passivation is mainly achieved through passivation materials that can undergo chemical reactions such as adsorption and precipitation with heavy metals. However, many passivators can cause new soil pollution, damage the soil structure, and even inhibit crop growth. Summary of the Invention

[0003] The object of the present disclosure is to provide a microbial inoculant including Alcaligenes balsami and Arthrobacter nicotianae, and a method for heavy metal passivation. The microbial inoculant of the present disclosure has a good passivation effect on heavy metals.

[0004] To achieve the above object, in a first aspect of the present disclosure, a microbial inoculant for heavy metal passivation is provided. The microbial inoculant includes a main inoculant and an auxiliary inoculant. The main inoculant includes Alcaligenes balsami; the auxiliary inoculant includes Arthrobacter nicotianae; The classification and naming of Alcaligenes balsami is Alcaligenes balsami Alcaligenes aquatilis , and its deposit number is CGMCC No. 14474; the classification and naming of Arthrobacter nicotianae is Arthrobacter nicotinovorans , and its deposit number is CGMCC No. 33646.

[0005] In the microbial inoculant, the viable bacteria number ratio of the main inoculant to the auxiliary inoculant is 1:(0.1 - 2).

[0006] Optionally, the viable bacteria number ratio of Alcaligenes balsami to Arthrobacter nicotianae is 1:(0.5 - 1.7).

[0007] Optionally, for the solid microbial inoculant, in the microbial inoculant, the viable count of Alcaligenes balsaminae is 10 5 to 10 12 CFU / g; or, for the liquid microbial inoculant, in the microbial inoculant, the viable count of Alcaligenes balsaminae is 10 5 to 10 12 CFU / mL.

[0008] Optionally, the co-inoculant further includes Phanerochaete chrysosporium; The taxonomic name of the Phanerochaete chrysosporium is Phanerochaete chrysosporium Phanerochaete chrysosporium and its deposit number is CGMCC No. 41170.

[0009] Optionally, the ratio of the viable count of Phanerochaete chrysosporium to the viable count of Arthrobacter nicotianae is 1:(1 - 4).

[0010] Optionally, the ratio of the viable count of Phanerochaete chrysosporium to the viable count of Arthrobacter nicotianae is 1:(1.5 - 3).

[0011] Optionally, for the solid microbial inoculant, in the microbial inoculant, the viable count of Alcaligenes balsaminae is 10 8 to 10 11 CFU / g; or, for the liquid microbial inoculant, in the microbial inoculant, the viable count of Alcaligenes balsaminae is 10 8 to 10 11 CFU / mL.

[0012] The second aspect of the present disclosure provides a method for heavy metal passivation, and the passivating agent used includes the microbial inoculant described in the first aspect of the present disclosure.

[0013] Optionally, the method includes: applying the microbial inoculant to the water body; Relative to each milliliter of the water body, the application amount of the microbial inoculant is 10 4 to 10 9 CFU.

[0014] Through the above technical solution, the present disclosure combines Achromobacter impatiens and Arthrobacter nicotianae to obtain a composite bactericide that can effectively passivate heavy metal ions. Using the microbial bactericide containing Achromobacter impatiens and Arthrobacter nicotianae of the present disclosure for the treatment of water and soil environments can effectively improve the acidity and alkalinity of water bodies and soils. At the same time, it has a significant passivation effect on common heavy metal ions in water and soil environments, reduces the content of available heavy metals in water and soil environments, increases the content of urease-producing bacteria in the soil, and increases the proportion of stable heavy metals, having an excellent effect on improving the quality of heavy metal-contaminated soil, and can achieve the efficient resource utilization of polluted environments such as water bodies and soils.

[0015] Other features and advantages of the present disclosure will be described in detail in the following specific implementation section.

[0016] Biological material preservation information Achromobacter impatiens of the present disclosure Alcaligenes aquatilis Is preserved in the China General Microbiological Culture Collection Center, preservation address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, postal code 100101, preservation date: July 31, 2017, and the strain preservation number is: CGMCC No. 14474.

[0017] Phanerochaete chrysosporium of the present disclosure Phanerochaete chrysosporium Is preserved in the China General Microbiological Culture Collection Center, preservation address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, postal code 100101, preservation date: February 26, 2024, and the strain preservation number is: CGMCC No. 41170.

[0018] Arthrobacter nicotianae of the present disclosure ( Arthrobacter nicotinovorans ) is preserved in the China General Microbiological Culture Collection Center, preservation address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, postal code 100101, preservation date: February 24, 2025, and the strain preservation number is: CGMCC No. 33646. Specific implementation mode

[0019] The following details the specific implementation mode of the present disclosure. It should be understood that the specific implementation mode described herein is only used to illustrate and explain the present disclosure, and is not used to limit the present disclosure.

[0020] The first aspect of the present disclosure provides a microbial bactericide for heavy metal passivation. The microbial bactericide includes a main bactericide and an auxiliary bactericide. The main bactericide includes Achromobacter impatiens; the auxiliary bactericide includes Arthrobacter nicotianae; The classification and naming of the said Alkaligenes impatiens is Alkaligenes impatiens Alcaligenes aquatilis , and its preservation number is CGMCC No.14474; the classification and naming of the said Arthrobacter nicotianae is Arthrobacter nicotinovorans , and its preservation number is CGMCC No.33646.

[0021] The inventors of the present disclosure compounded Alkaligenes impatiens and Arthrobacter nicotianae, and the obtained compound microbial agent can effectively passivate heavy metal ions. Using the microbial agent containing Alkaligenes impatiens and Arthrobacter nicotianae for the treatment of water and soil environment can significantly regulate the acid-base balance of aqueous solution and soil, and show an effective passivation effect on the heavy metal ions commonly present in the water and soil environment, reduce the content of available heavy metals in water and soil, so that the polluted water bodies and soil can be efficiently treated and recycled. It can also increase the number of urease-producing bacteria in the soil and at the same time increase the proportion of stable heavy metals in the soil.

[0022] According to an embodiment of the present disclosure, the said microbial agent is used to passivate heavy metals in soil and / or water.

[0023] According to an embodiment of the present disclosure, the dosage form of the said microbial agent includes liquid agent and / or solid agent. When the microbial agent is a solid agent, it can be directly applied or formulated into a liquid agent for application; the liquid agent can be directly applied after being cultured in a liquid medium, or formulated into a liquid microbial agent after cultivation.

[0024] According to an embodiment of the present disclosure, the preparation method of the strains used in the present disclosure can include: inoculating the strains into a culture medium for cultivation; the culture medium can be liquid or solid, and there is no particular limitation on the specific type. There is no particular limitation on the cultivation conditions of the strains, and common conditions can be used. For example, Alkaligenes impatiens is inoculated into LB liquid medium and cultured at 30-35°C with shaking for 1-3 days. Phanerochaete chrysosporium is inoculated into PDB liquid medium and cultured at 25-30°C with shaking for 2-4 days. Arthrobacter nicotianae is inoculated into LB liquid medium and cultured at 28-37°C with shaking for 1-3 days. During the cultivation process, the concentration of viable bacteria can be obtained by conventional methods, such as the hemocytometer counting method or the OD value observation method.

[0025] According to an embodiment of the present disclosure, the cultured microbial agent can be further processed into a microbial agent with a more convenient storage dosage form through steps including sterile filtration, freeze-drying, etc.

[0026] According to an embodiment of the present disclosure, in the microbial inoculant, the viable count ratio of the main inoculant to the auxiliary inoculant is 1:(0.1 - 2), preferably 1:(1.3 - 1.7), including but not limited to 1:0.1, 1:0.2, 1:0.5, 1:0.8, 1:1, 1:1.3, 1:1.5, 1:1.7, 1:2, or the range composed of any two of them; controlling the viable count ratio of the main inoculant and the auxiliary inoculant within the above range can improve the treatment effect on the water and soil environment, further reduce the heavy metal content, and increase the content of urease-producing bacteria in the soil.

[0027] According to an embodiment of the present disclosure, the viable count ratio of Alcaligenes impatiens to Arthrobacter nicotianae is 1:(0.5 - 1.7), including but not limited to 1:0.5, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.5, 1:1.7, or the range composed of any two of them, and the viable count of Alcaligenes impatiens is 10 5 to 10 12 CFU / g, preferably 10 8 to 10 11 CFU / g; or, the viable count of Alcaligenes impatiens is 10 5 to 10 12 CFU / mL, preferably 10 8 to 10 11 CFU / mL; compounding Alcaligenes impatiens and Arthrobacter nicotianae and controlling the viable count ratio within the above range, the obtained microbial inoculant can further improve the heavy metal passivation effect and increase the content of urease-producing bacteria in the soil.

[0028] According to an embodiment of the present disclosure, the microbial inoculant is a solid agent. In the microbial inoculant, the viable count of the main inoculant is 10 5 to 10 12 CFU / g, preferably 10 8 to 10 11 CFU / g, including but not limited to 10 5 CFU / g, 0.2×10 6 CFU / g, 0.5×10 6 CFU / g, 0.8×10 6 CFU / g, 10 6 CFU / g, 0.5×10 7 CFU / g, 10 7 CFU / g, 0.5×10 8 CFU / g, 10 8 CFU / g, 0.5×10 9 CFU / g, 109 CFU / g, 0.5×10 10 CFU / g, 10 10 CFU / g, 0.5×10 11 CFU / g, 10 11 CFU / g, 0.5×10 12 CFU / g, 10 12 CFU / g, or the range composed of any two of them; or, the microbial inoculant is a liquid agent, and in the microbial inoculant, the viable count of the main inoculant is 10 5 to 10 12 CFU / mL, preferably 10 8 to 10 11 CFU / mL, including but not limited to 10 5 CFU / mL, 0.2×10 6 CFU / mL, 0.5×10 6 CFU / mL, 0.8×10 6 CFU / mL, 10 6 CFU / mL, 0.5×10 7 CFU / mL, 10 7 CFU / mL, 0.5×10 8 CFU / mL, 10 8 CFU / mL, 0.5×10 9 CFU / mL, 10 9 CFU / mL, 0.5×10 10 CFU / mL, 10 10 CFU / mL, 0.5×10 11 CFU / mL, 10 11 CFU / mL, 0.5×10 12 CFU / mL, 10 12 CFU / mL, or the range composed of any two of them.

[0029] According to an embodiment of the present disclosure, the auxiliary inoculant further includes the Phanerochaete chrysosporium, that is, the microbial inoculant includes the Alcaligenes aquilegiicola, the Phanerochaete chrysosporium, and the Arthrobacter nicotianae. The taxonomic name of the Phanerochaete chrysosporium is Phanerochaete chrysosporium, and the deposit number is CGMCC No. 41170.

[0030] According to an embodiment of the present disclosure, the ratio of the viable count of Phanerochaete chrysosporium to the viable count of Arthrobacter nicotianae is 1:(1 - 4), preferably 1:(1.5 - 3), including but not limited to 1:1, 1:1.5, 1:1.8, 1:2, 1:2.5, 1:3, 1:3.2, 1:3.5, 1:4, or the range composed of any two of them; further preferably, the microbial inoculant is a solid agent. In the microbial inoculant, the viable count of Alcaligenes balsaminae is 10 8 to 10 11 CFU / g, including but not limited to 10 8 CFU / g, 0.5×10 9 CFU / g, 10 9 CFU / g, 0.5×10 10 CFU / g, 10 10 CFU / g, 0.5×10 11 CFU / g, 10 11 CFU / g, or the range composed of any two of them; alternatively, the microbial inoculant is a liquid agent. In the microbial inoculant, the viable count of Alcaligenes balsaminae is 10 8 to 10 11 CFU / mL, including but not limited to 10 8 CFU / mL, 0.5×10 9 CFU / mL, 10 9 CFU / mL, 0.5×10 10 CFU / mL, 10 10 CFU / mL, 0.5×10 11 CFU / mL, 10 11 CFU / mL, or the range composed of any two of them; By compounding the three strains and controlling the viable count and ratio, it is beneficial to improve the heavy metal treatment effect on water and soil environments and increase the number of urease-producing bacteria in the soil.

[0031] According to an embodiment of the present disclosure, the microbial inoculant is a liquid microbial inoculant. The method for preparing the liquid microbial inoculant may include: mixing and compounding liquid inoculant A, liquid inoculant C, and optional liquid inoculant B. Liquid inoculant A contains Alcaligenes balsaminae CGMCC No.14474, liquid inoculant B contains Phanerochaete chrysosporium CGMCC No.41170, and liquid inoculant C contains Arthrobacter nicotianae CGMCC No.33646. Liquid inoculants A, B, and C respectively contain liquid culture medium or buffer solution, and there is no special limitation on the specific type.

[0032] According to an embodiment of the present disclosure, the microbial inoculant is a solid inoculant, and the preparation method may include mixing the cultured strains or loading them on a solid carrier, and the type of the solid carrier is not particularly limited.

[0033] The second aspect of the present disclosure provides a method for heavy metal passivation, and the passivating agent used includes the microbial inoculant described in the first aspect of the present disclosure.

[0034] According to an embodiment of the present disclosure, the method includes: applying the microbial inoculant to the soil; optionally, relative to each kilogram of soil, the application amount of the microbial inoculant is 10 4 to 10 9 CFU, preferably 10 5 to 10 8 CFU, including but not limited to 10 4 CFU, 0.2×10 5 CFU, 0.5×10 5 CFU, 10 5 CFU, 0.2×10 6 CFU, 0.5×10 6 CFU, 10 6 CFU, 0.2×10 7 CFU, 0.5×10 7 CFU, 10 7 CFU, 0.2×10 8 CFU, 0.5×10 8 CFU, 10 8 CFU, 0.2×10 9 CFU, 0.5×10 9 CFU, 10 9 CFU, or the range composed of any two of them; the above application amount is beneficial to improving the heavy metal passivation effect.

[0035] According to an embodiment of the present disclosure, the method includes: applying the microbial inoculant to the water body; optionally, relative to each milliliter of the water body, the application amount of the microbial inoculant is 10 4 to 10 9 CFU, preferably 10 5 to 10 8 CFU, including but not limited to 10 4 CFU, 0.2×10 5 CFU, 0.5×10 5 CFU, 10 5 CFU, 0.2×10 6 CFU, 0.5×10 6 CFU, 10 6 CFU, 0.2×107 CFU, 0.5×10 7 CFU, 10 7 CFU, 0.2×10 8 CFU, 0.5×10 8 CFU, 10 8 CFU, 0.2×10 9 CFU, 0.5×10 9 CFU, 10 9 CFU, or the range composed of any two of them; the above application amounts are beneficial to improving the heavy metal passivation effect.

[0036] According to an embodiment of the present disclosure, there are no special requirements for the application method of the microbial inoculant, and it can be applied together with the base fertilizer or top dressing, etc.

[0037] The present invention will be further described in detail below in conjunction with embodiments, but the scope of the present invention is not limited to the following embodiments.

[0038] Unless otherwise specified, all culture media used are commercially available products.

[0039] Test method for metal ion concentration in water body: Atomic Absorption Spectrometry (AAS) (ISO 15586, APHA3111).

[0040] Test method for metal ion concentrations of available, residual, and carbonate-bound forms in soil: Microwave digestion-Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) (EPA 6010, EPA 3050B pretreatment standards).

[0041] Test method for the number of urease-producing bacteria in soil: Dilution plating method (urea selective medium) ( "Analysis Methods of Soil Microbiology" (edited by Lu Rukun), ISO 14238).

[0042] Strain culture: (1) Inoculate Alcaligenes aquilegae CGMCC No.14474 into LB medium and culture it for 1 day under the condition of shaking culture at 35°C.

[0043] (2) Inoculate Phanerochaete chrysosporium CGMCC No.41170 into PDB medium and culture it for 3 days under the condition of shaking culture at 28°C.

[0044] (3) Inoculate Arthrobacter nicotianae CGMCC No.33646 into LB medium and culture it for 1 day under the condition of shaking culture at 35°C.

[0045] Culture medium formula: LB medium (1 L): Tryptone: 10 g; Yeast Extract: 5 g; Sodium chloride (NaCl): 10 g; Deionized water: make up to 1 L; Adjust the pH to 7.2 with 5 mol / L NaOH (about 0.2 mL), and sterilize at 121°C for 30 min.

[0046] PDB culture medium (1 L): Potato Infusion: 4 g (or filtrate of 200 g fresh potato boiled and extracted); Dextrose: 20 g; Deionized water: make up to 1 L.

[0047] Preparation method of the microbial agent used in the test example: After preparing the bacterial solution according to the above-mentioned strain culture method, the bacterial solutions are mixed in a certain proportion.

[0048] Test Example 1 Heavy Metal Tolerance Test 1. Materials and Reagents: Bacteria: Bacteria to be tested that have been activated to the logarithmic growth phase (OD 600 is 0.6); Culture medium: X1 (representing Alcaligenes sphaerocephalus CGMCC No.14474) and X3 (representing Arthrobacter nicotianae CGMCC No.33646) use LB culture medium, and X2 (representing Phanerochaete chrysosporium CGMCC No.41170) uses PDB culture medium (the components of the culture medium do not contain the target heavy metals); Heavy metal solution: prepare standard mother solutions (containing cadmium chloride, zinc chloride, and copper chloride) containing target metals (Cd²⁺, Zn²⁺, and Cu²⁺), sterilize and filter using a 0.22 μm filter membrane; 2. Experimental Procedure (1) According to the concentration gradient in Table 1, different heavy metal solutions containing different heavy metal concentrations were added to the culture medium as test solutions, and the control group was a culture medium without heavy metals.

[0049] (2) Add an equal amount of bacterial suspension (final concentration 1×10 6 CFU / mL), and cultured at 37°C, 180 rpm for 1-2 days. The turbidity was observed with the naked eye to determine whether the bacteria were tolerant to the corresponding heavy metal concentrations. In Table 1, √ indicates that the bacteria can grow on the culture dish with the corresponding heavy metal concentration, and × indicates that the bacteria cannot grow.

[0050] (3) Based on the above experimental results, the obtained heavy metal concentration sensitivity results were further verified by the dilution coating method.

[0051] The test results are listed in Table 1.

[0052] Table 1

[0053] As can be seen from the results in Table 1, the strains in the microbial inoculant of the present disclosure can grow under conditions of a relatively high heavy metal ion concentration and have strong metal tolerance performance.

[0054] Test Example 2 Aqueous solution experiment In the passivation experiment with the solution standing still (initial pH = 7.01), the initial conditions were as follows: the cadmium ion concentration was 10 mg / L -1 , the copper ion concentration was 400 mg / L -1 , and the zinc ion concentration was 800 mg / L -1 . Different microbial inoculants were added according to the treatment, and the application amount of the microbial inoculant was 10 7 CFU / mL. After shaking for 4 h, the supernatant was taken by centrifugation, and the concentrations of various metal ions in it were measured. The results are listed in Table 2.

[0055] Microbial inoculant A1: The viable count ratio X1:X3 = 1:1.5, the viable count ratio of the main inoculant to the auxiliary inoculant is 1:1.5, and the viable count of Alcaligenes aquamarina in the microbial inoculant is 0.4×10 10 CFU / mL; Microbial inoculant A2: The viable count ratio X1:X3 = 1:3, the viable count ratio of the main inoculant to the auxiliary inoculant is 1:3, and the viable count of Alcaligenes aquamarina in the microbial inoculant is 0.25×10 10 CFU / mL; Microbial inoculant A3: The viable count ratio X1:X2:X3 = 1:1:0.5, the viable count ratio of the main inoculant to the auxiliary inoculant is 1:1.5, X2:X3 = 1:0.5, and the viable count of Alcaligenes aquamarina in the microbial inoculant is 0.4×10 10 CFU / mL; Microbial inoculant A4: The viable count ratio X1:X2:X3 = 1:0.5:1, the viable count ratio of the main inoculant to the auxiliary inoculant is 1:1.5, X2:X3 = 1:2, and the viable count of Alcaligenes aquamarina in the microbial inoculant is 0.4×10 10 CFU / mL; Microbial inoculant A5: The viable count ratio X1:X2:X3 = 1:0.75:0.75, the viable count ratio of the main inoculant to the auxiliary inoculant is 1:1.5, X2:X3 = 1:1, and the viable count of Alcaligenes aquamarina in the microbial inoculant is 0.4×10 10 CFU / mL; Microbial inoculant A6: X1, and the viable count of Alcaligenes aquamarina in the microbial inoculant is 10 10CFU / mL; Microbial inoculant A7: A heavy metal contaminated soil remediation microbial inoculant product produced by Shandong Maikezhen Biotechnology Co., Ltd., with viable count of 10 10 CFU / mL; Microbial inoculant A8: The viable count ratio of X2:X3 = 1:0.5. The viable count of the auxiliary microbial agent in the microbial inoculant is 10 10 CFU / mL, and the viable count of the main microbial agent is 0; Microbial inoculant A9: The viable count ratio of X1:X33 = 1:1.5. The viable count of Alcaligenes aquamarina in the microbial inoculant is 0.4×10 10 CFU / mL; X33 is other Arthrobacter nicotianae screened in the same batch; Microbial inoculant A10: The viable count ratio of X11:X3 = 1:1.5. The viable count of Alcaligenes aquamarina in the microbial inoculant is 0.4×10 10 CFU / mL; X11 is other Alcaligenes aquamarina screened in the same batch.

[0056] Table 2

[0057] According to the data in Table 2, it can be seen that using the microbial inoculant of the present disclosure to treat the water body containing heavy metal ions can significantly reduce the heavy metal content and at the same time increase the pH value. Further, according to the comparison between microbial inoculants A1 and A2, when the viable count ratio of the main microbial agent to the auxiliary microbial agent is within the range of 1:(0.1 - 2), the heavy metal treatment effect can be improved; according to the comparison between microbial inoculants A3 - A5 and A1 - A2, when the microbial inoculant contains Alcaligenes aquamarina, Phanerochaete chrysosporium and Arthrobacter nicotianae, the heavy metal passivation effect is better; according to the comparison among microbial inoculants A3 - A5, when the ratio of the viable count of Phanerochaete chrysosporium to the viable count of Arthrobacter nicotianae is within the range of 1:(1 - 4), preferably within the range of 1:(1.5 - 3), the heavy metal passivation rate is higher.

[0058] Test Example 3 Soil Experiment Based on the soil environment for wheat growth, the cadmium ion concentration is 8 mg L -1 , and the available state concentration is 6.80 mg L -1 ; the copper ion concentration is 50 mg L -1 , and the available state concentration is 42.3 mg L -1 ; the zinc ion concentration is 90 mg L -1 , and the available state concentration is 81.5 mg L -1 . Different microbial inoculants were added according to the treatment, and the application amount of the microbial inoculant was 10 10CFU / kg, a 45-day pure soil cultivation experiment was carried out, and the test results are listed in Table 3.

[0059] Table 3

[0060] According to the data in Table 3, it can be seen that the microbial inoculant of the present disclosure can increase the pH value of the soil, reduce the contents of available Cd ions, Cu ions and Zn ions in the soil, and at the same time increase the contents of carbonate-bound and residual forms in the soil. The number of urease-producing bacteria in the soil also increases, which is beneficial to the alkaline passivation of heavy metal ions in the soil. Further, according to the comparison between microbial inoculants A1 and A2, when the ratio of the viable cell numbers of Alcaligenes aquatilis and Arthrobacter nicotianae is in the range of 1:(0.1 - 2), the heavy metal treatment effect can be improved; according to the comparison between microbial inoculants A3 - A5 and A1 - A2, when the microbial inoculant contains Alcaligenes aquatilis, Phanerochaete chrysosporium and Arthrobacter nicotianae, the heavy metal passivation effect is better; according to the comparison among microbial inoculants A3 - A5, when the ratio of the viable cell number of Phanerochaete chrysosporium to the viable cell number of Arthrobacter nicotianae is in the range of 1:(1 - 4), preferably in the range of 1:(1.5 - 3), the reduction rate of heavy metals and the number of urease-producing bacteria are higher.

[0061] The preferred embodiments of the present disclosure have been described in detail above. However, the present disclosure is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present disclosure, various simple modifications can be made to the technical solutions of the present disclosure, and these simple modifications all belong to the protection scope of the present disclosure.

[0062] In addition, it should be noted that, in the case of no contradiction, the various specific technical features described in the above specific embodiments can be combined in any suitable way. To avoid unnecessary repetition, the present disclosure does not separately describe various possible combination ways.

[0063] In addition, any combination can be made between various different embodiments of the present disclosure, as long as it does not violate the idea of the present disclosure, and it should also be regarded as the content disclosed by the present disclosure.

Claims

1. A microbial inoculant for heavy metal passivation, characterized in that, The microbial agent includes a main agent and an auxiliary agent, wherein the main agent includes Alcaligenes sphaerocephalus; and the auxiliary agent includes Arthrobacter nicotianae; The classification and naming of the Alkaligenes impatiens is Alkaligenes impatiens Alcaligenes aquatilis , and its preservation number is CGMCC No. 14474; the classification and naming of the Arthrobacter nicotianae is Arthrobacter nicotinovorans , and its preservation number is CGMCC No. 33646.

2. The microbial inoculum according to claim 1, wherein In the microbial agent, the ratio of the number of live bacteria of the main agent to that of the auxiliary agent is 1:(0.1-2).

3. The microbial inoculant according to claim 2, wherein, The ratio of the live bacteria count of the water balsam pear alcaligenes and the tobacco balsam pear Arthrobacter is 1: (0.5-1.7).

4. The microbial inoculant according to claim 3, wherein The solid microbial inoculant. In the microbial inoculant, the viable count of the Alcaligenes impatiens is 10 5 to 10 12 CFU / g; or, The microbial inoculum is a liquid agent. In the microbial inoculum, the viable count of the Alcaligenes balsaminae is 10 5 to 10 12 CFU / mL.

5. The microbial inoculant according to claim 2, wherein, The auxiliary bacterial agent also includes Phanerochaete chrysosporium; The taxonomic name of the Phanerochaete chrysosporium is Phanerochaete chrysosporium Phanerochaete chrysosporium , and the preservation number is CGMCC No. 41170.

6. The microbial inoculum according to claim 5, wherein, The ratio of the number of live bacteria of Phanerochaete chrysosporium to the number of live bacteria of Arthrobacter nicotianae is 1:(1-4).

7. The microbial inoculant according to claim 5, wherein The ratio of the number of live bacteria of Phanerochaete chrysosporium to the number of live bacteria of Arthrobacter nicotianae is 1:(1.5-3).

8. The microbial inoculum according to claim 6 or 7, wherein The solid microbial inoculant. In the microbial inoculant, the viable count of the Alcaligenes balsaminae is 10 8 to 10 11 CFU / g; or, The microbial inoculant is a liquid agent. In the microbial inoculant, the viable count of the Alcaligenes balsaminae is 10 8 to 10 11 CFU / mL.

9. A method for passivating heavy metals, characterized in that, The passivating agent used includes the microbial agent described in any one of claims 1 to 8.

10. The method according to claim 9, wherein The method comprises: applying the microbial agent to a water body; The application amount of the microbial inoculum is 10 4 to 10 9 CFU per milliliter of the water body.

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