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Composite bacterial agent, preparation method thereof and application in field of soil remediation

A technology of compound inoculum and bacillus, which is applied in the preparation of compound inoculants and in the field of compound microbial inoculants, can solve problems such as long-term stability of unfavorable soils

Active Publication Date: 2020-07-17
SHANDONG BEE LAN BIOTECHNOLOGY CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The inventor believes that the introduction of chemical components for soil remediation is likely to bring new environmental problems, which may not be conducive to the long-term stability of the soil

Method used

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  • Composite bacterial agent, preparation method thereof and application in field of soil remediation
  • Composite bacterial agent, preparation method thereof and application in field of soil remediation
  • Composite bacterial agent, preparation method thereof and application in field of soil remediation

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preparation example Construction

[0048] The second aspect of the present invention provides the preparation method of the composite bacterial agent described in the first aspect, the preparation method comprising the following steps: Bacillus tianshenli strain S5, Bacillus tianshenli P46, Bacillus tianshenli strain Bacillus velezensis S3 and Bacillus subtilis P30 are fermented separately to obtain fermentation liquid, and the fermentation liquid is dried to obtain bacterial powder, which is obtained by compounding the bacterial powder of the above strains with the substrate.

[0049] Preferably, the fermentation includes slant culture, primary seed culture, expanded culture and fermenter culture, and when the spore rate in the fermenter reaches more than 90%, the fermentation broth is collected.

[0050] Further preferably, the slant culture is cultured at 37°C for 24-30 hours.

[0051] Further preferably, the specific method of the primary seed cultivation is as follows: use an inoculation loop to inoculate ...

Embodiment 1

[0061] Embodiment one, the screening of strain

[0062] 1. Materials and methods

[0063] 1.1 Samples: 11 sample materials such as soil and water samples around the electroplating plant.

[0064] 1.2 Screening medium:

[0065] Nutrient medium for bacterial strains: NB synthetic medium, 1.5% agar is added on the basis of NB medium to form a solid medium.

[0066] Screening medium (NBRIP) of phospholytic bacteria strains: glucose 1.0%, Ca 3 (PO 4 ) 2 0.5%, MgCl 2 0.5%, peptone 0.5%, agar powder 2.0%, pH 7.2-7.4, sterilized at 121°C for 30 minutes.

[0067] Screening medium for sulfate-reducing bacteria strains: dipotassium hydrogen phosphate 0.05%, anhydrous calcium chloride 0.01%, ammonium chloride 0.1%, magnesium sulfate heptahydrate 0.2%, ferrous sulfate heptahydrate 0.05%, sodium chloride 0.1 %, 0.05% ascorbic acid, 0.05% L-Cys cysteine, 0.5% anhydrous sodium sulfate, 0.65% sodium lactate solution, 2.0% agar powder, pH 6.8-7.0, sterilized at 121°C for 30min.

[006...

Embodiment 2

[0084] Embodiment two, identification and safety test of bacterial classification

[0085] 1. Materials and methods

[0086] 1.1 Bacterial strains: The microorganisms isolated from the soil and water samples of the electroplating plant were screened through primary screening and secondary screening, and a total of 4 strains with high heavy metal degradation performance were screened out, and the numbers were P30, P46, S3 and S5.

[0087] 1.2 Morphological identification and molecular identification of bacteria: Unless otherwise specified, general morphological and molecular tests were carried out with reference to "Bergey's Bacterial Identification Manual" (9th edition) and "Microbiology Experimental Manual".

[0088] 1.3 Safety test: The phytotoxicity test of the bacteria is carried out by using "pearl" tomato seeds and "crystal" lettuce seeds. Seeds were soaked in 2% concentration of single bacterial solution, mixed bacterial solution (1:1:1) and distilled water (control gr...

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Abstract

The invention belongs to the technical field of soil heavy metal remediation, and specifically relates to a composite bacterial agent, a preparation method thereof and an application in the field of soil remediation. The invention provides 2 strains of bacillus tianshenli having soil heavy metal remediation effects, 1 strain of bacillus subtilis, and 1 strain of bacillus velezensis. The strains provided by the invention are obtained beneficial strains that efficiently adsorb soil heavy metals including cadmium and lead and are screened from materials such as soil, water samples and plants around an electroplating plant through technologies such as screening, purification and identification, safety tests are performed on the strains, fermentation conditions are studied, compound compatibility is performed, and after verification of potted and field experiments, the novel microecological preparation-soil heavy metal remediation bacteria agent is provided.

Description

technical field [0001] The invention belongs to the technical field of soil heavy metal remediation, and in particular relates to a method composed of Bacillus tianshenli strain S5, Bacillus tianshenli P46, Bacillus velezensis S3, Bacillus subtilis A composite microbial agent composed of P30, a preparation method of the composite microbial agent, and an application of the composite microbial agent in the field of soil heavy metal remediation. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily to be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Heavy metal pollution in farmland mainly comes from mining waste residues, pesticides, wastewater, sludge, and atmospheric deposition. Excessive heavy metal deposition can lead...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/07C12R1/125
CPCB09C1/10C12N1/20C12N1/205C12R2001/07C12R2001/125
Inventor 单宝龙胡著然郭静王丽荣刘晓晓孔德荣张文娟申小冉朱自然董现义
Owner SHANDONG BEE LAN BIOTECHNOLOGY CO LTD
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