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CN-122012271-A - Composite microbial agent for treating mine tailing polluted soil and preparation method and application thereof

CN122012271ACN 122012271 ACN122012271 ACN 122012271ACN-122012271-A

Abstract

The invention belongs to the technical field of soil microbial control, and discloses a composite microbial agent for treating mine tailing polluted soil, which comprises bacterial liquid containing Coxsackie bacteria P1-4 and bacterial liquid containing stenotrophomonas R5 in a volume ratio of 1-2:1-2, wherein the OD600 values of the bacterial liquid and the bacterial liquid are independently 0.5-1.5. The invention also discloses a preparation method of the composite microbial inoculum and application of the composite microbial inoculum in treating mine tailing polluted soil. The screened kosakazakii P1-4 and the oligotrophic monad R5 are both derived from cadmium-resistant environments (rhizosphere of miscanthus sinensis and endophyte of rice), have natural tolerance to heavy metal stress, and the microbial agent prepared by compounding the same has extremely strong adaptability and reparative pertinence to typical acidic and cadmium-lead composite polluted soil in mine tailing areas, and solves the common problems of low survival rate and functional failure of common microbial agents in severe tailing environments.

Inventors

  • HU RUIWEN
  • LI JUAN
  • Tian Yunhe
  • QIN TIAN
  • Tian meijie
  • CHEN XINYU

Assignees

  • 湖南农业大学

Dates

Publication Date
20260512
Application Date
20260107

Claims (10)

  1. 1. The composite microbial agent for treating mine tailing polluted soil is characterized by comprising bacterial liquid containing Coxsackie bacteria (Kosakonia sacchari) P1-4 and bacterial liquid containing Stenotrophomonas (Stenotrophomonas sp.) R5 in a volume ratio of 1-2:1-2, wherein the OD600 value of the bacterial liquid containing Coxsackie bacteria (Kosakonia sacchari) P1-4 and the bacterial liquid containing Stenotrophomonas (Stenotrophomonas sp.) R5 is independently 0.5-1.5; Wherein, the Coasa bacteria (Kosakonia sacchari) P1-4 is preserved in China center for type culture Collection, the preservation date is 2023, 09 and 20 days, the preservation address is China university of Wuhan, and the preservation number is CCTCC NO: M20231759; Stenotrophomonas (Stenotrophomonas sp.) R5 is deposited with China center for type culture Collection with a date of 20 days of 09 months in 2023, and with a deposit address of university of Wuhan, china with a deposit number of CCTCC NO: M20231758.
  2. 2. The preparation method of the compound microbial agent for treating mine tailing polluted soil is characterized by comprising the following steps of: (1) Inoculating the Coxsackie bacteria (Kosakonia sacchari) P1-4 into an LB culture medium, shake culturing, centrifuging after culturing, removing supernatant, and adjusting the OD600 value of the thalli to be 0.5-1.5 to obtain a bacterial liquid containing the Coxsackie bacteria (Kosakonia sacchari) P1-4; (2) Inoculating Stenotrophomonas (Stenotrophomonas sp.) R5 into an LB culture medium, shake culturing, centrifuging after culturing, removing supernatant, and adjusting the OD600 value of the thalli to be 0.5-1.5 to obtain a bacterial liquid containing Stenotrophomonas (Stenotrophomonas sp.) R5; (3) Mixing a bacterial solution containing the Coxsackie bacteria (Kosakonia sacchari) P1-4 and a bacterial solution containing the Stenotrophomonas (Stenotrophomonas sp.) R5 according to a volume ratio of 1-2:1-2 to obtain the compound microbial agent.
  3. 3. The method according to claim 2, wherein in the step (1), the condition of shake culture includes a culture temperature of 26 ℃ to 30 ℃, a culture time of 10 hours to 14 hours, and a rotation speed of 180rpm to 220rpm.
  4. 4. The method according to claim 2, wherein in the step (1), the rotational speed of centrifugation is 5800rpm to 6200rpm, and the time of centrifugation is 2min to 4min.
  5. 5. The method according to claim 2, wherein in the step (2), the condition of shake culture includes a culture temperature of 26 ℃ to 30 ℃, a culture time of 10 hours to 14 hours, and a rotation speed of 180rpm to 220rpm.
  6. 6. The method according to claim 2, wherein in the step (2), the rotational speed of centrifugation is 5800rpm to 6200rpm, and the time of centrifugation is 2min to 4min.
  7. 7. The application of the composite microbial agent of claim 1 or the composite microbial agent prepared by the preparation method of any one of claims 2-6 in mine tailing contaminated soil treatment.
  8. 8. The use according to claim 7, wherein 8-12 l of the composite microbial agent is applied to each cubic meter of mine tailing contaminated soil.
  9. 9. The use according to claim 7, wherein the mine tailing contaminated soil is mainly cadmium or/and lead contaminated soil.
  10. 10. The use according to claim 7, wherein the pH value of the mine tailing polluted soil is 4-4.5, the content of effective state cadmium is 0.5-0.7 mg/kg, the content of effective state lead is 7-9 mg/kg, and the content of organic matters is 5-7 mg/kg.

Description

Composite microbial agent for treating mine tailing polluted soil and preparation method and application thereof Technical Field The invention belongs to the technical field of soil microbial control, and particularly relates to a composite microbial agent for treating mine tailing polluted soil, and a preparation method and application thereof. Background The tailings produced in the mining process contain a large amount of heavy metals (such as cadmium, lead and the like) and other harmful substances, so that serious pollution is caused to the ecological environment. To combat such contamination, it has traditionally been largely dependent on physical and chemical methods, such as: The physical method comprises the steps of soil dressing, soil replacement, isolated landfill and the like. Although the methods can take effect rapidly, the engineering quantity is large, the cost is high, the method is only suitable for small-scale pollution, the pollutants are not eliminated fundamentally, and the risks of secondary treatment and pollution transfer exist. Chemical methods such as chemical immobilization/stabilization, rinsing, redox, etc. The method changes the form or the solubility of heavy metals by adding chemical reagents, and also has the problems of high treatment cost, possibility of damaging the soil structure, secondary pollution caused by introducing new chemical substances, uncertain long-term stability and the like. In view of the limitations of the conventional methods, bioremediation using microorganisms has been receiving attention in recent years because of their environmental friendliness, relatively low cost, mild repair process, ecological sustainability and the like. However, although the microbial remediation technology has wide prospects, the microbial remediation technology is limited by the characteristics of microorganisms and the complexity of the tailing environment in the practical application of the complex tailing polluted soil, most of the microbial remediation technology adopts microorganisms for remediation, and has limited remediation effect and low efficiency. Therefore, development of a microbial agent to improve the treatment effect is important. Disclosure of Invention The invention aims to solve the technical problems and overcome the defects and shortcomings in the background art, and provides a composite microbial agent for treating mine tailing polluted soil, and a preparation method and application thereof. In order to solve the technical problems, the technical scheme provided by the invention is as follows: The composite microbial agent for treating mine tailing polluted soil comprises bacterial liquid containing Coxsackie bacteria (Kosakonia sacchari) P1-4 and bacterial liquid containing Stenotrophomonas (Stenotrophomonas sp.) R5 in a volume ratio of 1-2:1-2, wherein the OD600 value of the bacterial liquid containing Coxsackie bacteria (Kosakonia sacchari) P1-4 and the bacterial liquid containing Stenotrophomonas (Stenotrophomonas sp.) R5 are independently 0.5-1.5; Wherein, the Coasa bacteria (Kosakonia sacchari) P1-4 is preserved in China center for type culture Collection, the preservation date is 2023, 09 and 20 days, the preservation address is China university of Wuhan, and the preservation number is CCTCC NO: M20231759; Stenotrophomonas (Stenotrophomonas sp.) R5 is deposited with China center for type culture Collection with a date of 20 days of 09 months in 2023, and with a deposit address of university of Wuhan, china with a deposit number of CCTCC NO: M20231758. Based on a general inventive concept, the invention also provides a preparation method of the compound microorganism bacterial agent for treating mine tailing polluted soil, which comprises the following steps: (1) Inoculating the Coxsackie bacteria (Kosakonia sacchari) P1-4 into an LB culture medium, shake culturing, centrifuging after culturing, removing supernatant, and adjusting the OD600 value of the thalli to be 0.5-1.5 to obtain a bacterial liquid containing the Coxsackie bacteria (Kosakonia sacchari) P1-4; (2) Inoculating Stenotrophomonas (Stenotrophomonas sp.) R5 into an LB culture medium, shake culturing, centrifuging after culturing, removing supernatant, and adjusting the OD600 value of the thalli to be 0.5-1.5 to obtain a bacterial liquid containing Stenotrophomonas (Stenotrophomonas sp.) R5; (3) Mixing a bacterial solution containing the Coxsackie bacteria (Kosakonia sacchari) P1-4 and a bacterial solution containing the Stenotrophomonas (Stenotrophomonas sp.) R5 according to a volume ratio of 1-2:1-2 to obtain the compound microbial agent. In the preparation method, preferably, in the step (1), the shake culture condition includes a culture temperature of 26 ℃ to 30 ℃, a culture time of 10 hours to 14 hours and a rotation speed of 180rpm to 220rpm. The temperature of the shake culture is more preferably 28 ℃, the time of the shake culture is more preferably 12 hours, and the OD600