Bacillus with aerobic arsenic methylation and volatilization functions and application thereof

A bacillus and methylation technology, applied in the field of microorganisms, can solve the problems of lack of improvement and promotion of nutrients and growth, unfavorable wide application, low arsenic concentration, etc., and achieve the effect of good plant growth promotion effect.

Active Publication Date: 2022-04-29
GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CN201510956127.5 identified an aerobic arsenic-methylating bacterium SM-1 of Cellulophilaceae, but the bacteria can only grow in R2A medium, and cannot be produced in LB and other medium, which has obvious limitations
At the same time, the arsenic concentration of the bacteria is low, which is not conducive to wide application.
In addition, the strains protected by the above-mentioned intellectual property rights only have the function of arsenic methylation, and do not improve or promote crop nutrients and growth, so the function is relatively single

Method used

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  • Bacillus with aerobic arsenic methylation and volatilization functions and application thereof
  • Bacillus with aerobic arsenic methylation and volatilization functions and application thereof
  • Bacillus with aerobic arsenic methylation and volatilization functions and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Isolation and Identification of Arsenic Methylation Functional Bacillus LH14

[0041] The samples were collected from arsenic-contaminated paddy soil in Lianhuashan, Shantou City, Guangdong Province. Weigh 100 g of soil samples that have been naturally air-dried and passed through a 10-mesh sieve into a 250-ml conical flask, add 100 ml of sterilized ultrapure water, seal the bottle mouth with a breathable membrane, and let it stand for 15 days at room temperature to activate soil microorganisms. The soil suspension was serially diluted with sterilized ultrapure water with a concentration gradient of 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 and 10 -7 , respectively take 100 μl concentration as 10 -4 , 10 -5 , 10 -6 and 10 -7 A dilution of 20 μM arsenite in ST10 -1 on solid medium plates. ST10 -1 The formula of the solid medium is as follows: tryptone 0.5g, yeast extract 0.05g, glucose 0.5g, dilute to 1000ml, and sterilize at 121°C for 20min und...

Embodiment 2

[0050] Example 2: Arsenic methylation efficiency of arsenic methylation functional Bacillus LH14 in arsenic-containing R2A, LB and TGY medium

[0051] LH14 was expanded and cultured in LB liquid medium for the verification of arsenic methylation efficiency. The details are as follows: prepare 500ml of sterilized LB liquid medium, the formula is 10g / L peptone, 5g / L yeast powder, 10g / L NaCl, and the balance is water; inoculate a single colony in the streaked plate into the medium for shaking culture (30°C, 200rpm) for 10 hours to 12 hours, its OD600 is 0.4-0.5, which is in the middle of the logarithmic phase. The expanded cultured bacterial solution was centrifuged at 5000rpm for 10min to collect the bacterial cells. Take 0.55ml LH14 bacteria solution (OD600=1.8) and inoculate them in 100ml sterile R2A, LB and TGY liquid medium containing 12μM arsenous acid respectively. The formula of TGY liquid medium is 5g / L tryptone, 5g / L yeast powder, 1g / L glucose, 1g / L K 2 HPO 4 , and ...

Embodiment 3

[0053] Example 3: Analysis of the growth curve of arsenic methylation-functional Bacillus LH14 in LB medium containing different arsenic concentrations.

[0054] The strains were inoculated in LB medium containing 0, 20, and 50 μM arsenous acid, and cultured for 5 days; at different cultivation time points, appropriate amount of culture was added to a centrifuge tube, centrifuged at 12,000 rpm for 2 minutes, and filtered through 0.22 μm. After membrane filtration of the supernatant, arsenic speciation was performed by HPLC-ICP-MS.

[0055] like Image 6 As shown, the strain LH14 in the culture medium with different concentrations of As(III) concentration, the growth lag phase of the arsenic-methylating functional Bacillus LH14 increases with the increase of As(III) concentration, and the time required for its growth to the logarithmic phase Increased from 14 hours to 30 hours. However, after culturing for 50 hours, the biomass was cultured under the condition of 50 μM arseno...

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Abstract

The invention discloses bacillus with aerobic arsenic methylation and volatilization functions and application thereof, and belongs to the technical field of microorganisms. The name of the bacterial strain is bacillus subtilis LH14, the bacterial strain is preserved in Guangdong Microbial Culture Collection Center (GDMCC) on December 2, 2021 in No.59 building, No.100 Courtyard, Xianlie Middle Road, Guangzhou City, Guangdong Province, and the preservation number is GDMCC No: 62092. The bacillus LH14 has an obvious methylation effect on inorganic trivalent arsenic, a main methyl arsenic product is trimethyl arsenic, a secondary methyl arsenic product is dimethyl arsenic, and double detoxification functions of arsenic methylation and arsenic oxidation can be synchronously realized. The bacillus LH14 can obviously reduce the content of highly toxic inorganic arsenic, generates organic arsenic with low toxicity and volatile arsenic, and has important application potential in treatment and remediation of arsenic polluted environment, especially treatment and remediation of arsenic polluted soil.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a bacillus with aerobic arsenic methylation and volatilization functions and application thereof. Background technique [0002] Long-term production activities such as arsenic mining, arsenic smelting, pesticide and fertilizer application, and sewage irrigation have enriched metalloid arsenic in the soil environment, causing soil arsenic pollution. Because rice plants are easy to absorb and accumulate arsenic, rice has become the main way for the human body to be chronically exposed to arsenic, which poses a serious threat to human health. Seasonal water management in paddy fields alters soil redox potential, accompanied by significant changes in arsenic speciation and mobility. Arsenic is mainly pentavalent arsenate under dry conditions in paddy fields, which is strongly adsorbed on the surface of oxides such as iron and aluminum, which limits the mobility of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C02F3/34A01N63/22A01P21/00C12R1/07C02F101/20
CPCC12N1/20B09C1/10C02F3/34A01N63/22C02F2101/20
Inventor 李芳柏方利平陈冠虹
Owner GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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