Bacillus megaterium, preparation of microbial inoculum thereof and application thereof in soil heavy metal remediation

A technology of Bacillus megaterium and heavy metals, applied in the field of bioengineering, can solve problems such as crop yield reduction, threats to human health, and harm to the normal growth of crops, and achieve the effects of promoting growth, realizing production-side repair, and preventing non-point source pollution

Active Publication Date: 2019-12-31
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Not only directly endanger the normal growth of crops, resulting in crop yield reduction, but also pollution factors can easily enter the human body through the food chain, seriously threatening human health

Method used

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  • Bacillus megaterium, preparation of microbial inoculum thereof and application thereof in soil heavy metal remediation
  • Bacillus megaterium, preparation of microbial inoculum thereof and application thereof in soil heavy metal remediation
  • Bacillus megaterium, preparation of microbial inoculum thereof and application thereof in soil heavy metal remediation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Screening of Bacillus megaterium NCT-5

[0033] This embodiment includes the following steps:

[0034] ① Cultivate Bacillus megaterium CGMCC NO.4698 preserved in the laboratory in a liquid medium (beef extract 3g / L, peptone 5g / L, NaCl 0.5g / L), 30°C, 200r / min shaker conditions for 12h, Use a hemocytometer to measure the concentration of effective viable bacteria, and dilute to prepare 10 8 CFU / mL bacterial suspension.

[0035] ②A 30W ultraviolet lamp was used for mutagenesis of the strain. Turn on the UV lamp before irradiation and preheat for 20 minutes. Use a pipette gun to draw 5mL of bacterial suspension into a 9cm sterile petri dish, adjust the distance between the UV lamp (30W) and the petri dish to 30cm (vertical distance), open the dish lid, and treat for 30s, 60s, 90s, 120s respectively. After mutagenesis, pipette 0.2 mL of bacterial suspension to coat the plate (KNO 3 4g / L, MgSO 4 ·7H 2 O 0.5g / L, KH 2 PO 4 0.5g / L, KCl 1g / L, CaCl 2 1mg / L, FeSO 4 ·7H...

Embodiment 2

[0043] The present embodiment relates to making the Bacillus megaterium (Bacillus megaterium) NCT-5 obtained in Example 1 into a solid bacterial agent, specifically comprising:

[0044] Seed medium: glucose 5g / L, beef extract 3g / L, peptone 5g / L, MgSO 4 ·7H 2 O 0.3g / L, use NaOH solution to adjust the pH to 6.5.

[0045] Spore-forming medium: bran 15g / L, potassium nitrate 10g / L, potassium dihydrogen phosphate 0.5g / L, magnesium sulfate 0.5g / L, manganese sulfate 0.1g / L, calcium carbonate 0.1g / L, using NaOH solution Adjust the pH to 7.0.

[0046] Cultivate Bacillus megaterium (Bacillus megaterium) NCT-5 (CGMCC NO.12910) in the seed medium, 30°C, 200r / min shaker conditions for 12h, and the seed solution is 2% (v / v) of the seed amount Inoculate in a 1000L fermenter pre-installed with 600L sterilized fermentation medium for cultivation, culture conditions: 37°C, 100r / min, ventilation rate 1.5L / min, no need to adjust pH during fermentation. After 10 hours of fermentation, the numbe...

Embodiment 3

[0063] The present embodiment relates to carrying out the remediation of secondary salinized soil with Bacillus megaterium (Bacillus megaterium) NCT-5 that embodiment 1 obtains, specifically comprising:

[0064] Seed medium: glucose 5g / L, beef extract 3g / L, peptone 5g / L, MgSO 4 ·7H 2 O 0.3g / L, use NaOH solution to adjust the pH to 6.5.

[0065] Spore-forming medium: corn flour 5g / L, molasses 10g / L, urea 10g / L, potassium dihydrogen phosphate 0.5g / L, magnesium sulfate 0.5g / L, manganese sulfate 0.1g / L, calcium carbonate 0.1g / L , using NaOH solution to adjust the pH to 7.0.

[0066] Step 1. Cultivate Bacillus megaterium (Bacillus megaterium) NCT-5 (CGMCC NO.12910) in the seed culture medium, 30°C, 200r / min shaker conditions for 12h, and the seed liquid was mixed with 2% (v / v) The seeds were inoculated in a 10L fermenter pre-installed with 7L sterilized fermentation medium for cultivation. The cultivation conditions were: 37°C, 300r / min, and the ventilation rate was 2.5L / min. Th...

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Abstract

The invention discloses bacillus megatherium, preparation of a microbial inoculum of the bacillus megatherium and an application thereof in soil heavy metal remediation. The invention relates to bacillus megaterium NCT-5 CGMCC NO.12910, and by applying a strain solid preparation provided by the invention, the nitrate content and the heavy metal content of soil can be effectively reduced. The method is simple and convenient to prepare, convenient to operate and low in treatment cost and can be widely applied to the field of soil remediation.

Description

technical field [0001] The invention relates to a technology in the field of bioengineering, in particular to the preparation of bacillus megaterium and its bacterial agent and its application in soil heavy metal restoration. Background technique [0002] In recent years, my country's protected vegetable industry has developed rapidly, but the particularity of the protected cultivation environment and unscientific management make the production of protected vegetables highly dependent on chemical fertilizers, compound fertilizers and other fertilizers, resulting in unbalanced soil nutrients, secondary salinization of soil and many other production processes. The problem arises. In particular, soil acidification will increase the mobility of heavy metals in the soil, and then intensify the accumulation of heavy metals in the soil. Among them, the pollution of Cd, Pb, and Cu caused by agrochemicals is the most prominent. In the actual production process, due to differences in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/01C12N13/00B09C1/10C12R1/11
CPCB09C1/105C12N1/20C12N13/00C12N15/01C12N1/205C12R2001/11
Inventor 张丹周培支月娥蒋淼王军才陈寻峰
Owner SHANGHAI JIAO TONG UNIV
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