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Acid-resistant denitrifying Bacillus subtilis strain, and composite microbial inoculant and preparation method thereof

A technology of compound microbial agent and Bacillus subtilis, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, chemical instruments and methods, etc., can solve the problems of not being able to adapt to low pH environment, heavy metal pollution, soil re-acidification, etc. Achieve the effects of improving soil biological fertility, improving biological activity, improving pH value and salt accumulation

Active Publication Date: 2016-06-01
山东黄河谷农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods have many advantages, they also have great disadvantages: irrigation and salt washing will wash the nitrate nitrogen into the ground and pollute the groundwater; the cost of the soil amendment method is relatively high, and chemical raw materials or industrial waste are mostly used as Raw materials have also caused problems such as soil re-acidification and heavy metal pollution.
These aerobic denitrifying Bacillus subtilis strains cannot adapt to the low pH environment of the northern greenhouse soil (pH as low as 4.0)

Method used

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  • Acid-resistant denitrifying Bacillus subtilis strain, and composite microbial inoculant and preparation method thereof
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  • Acid-resistant denitrifying Bacillus subtilis strain, and composite microbial inoculant and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: The Bacillus subtilis strain KD203-3 resistant to acid denitrification is obtained from soil screening and mutagenesis selection by the following steps:

[0030] 1. Isolation and purification of denitrification starting strain

[0031] Linzi District, Zibo City was selected as a greenhouse for planting for 10 years, and about 300g of deep (5-15cm) soil was taken. Weigh 10 g of freshly collected soil and put it into a conical flask (with several glass beads) filled with 90 mL of sterile water to make a dilution of 10 -1 soil suspension. Take another 7 test tubes filled with 9mL sterile water and take a sterile pipette, and make a dilution of 10 -4 →10 -5 →10 -6 →10 -7 →10 -8 Soil dilution.

[0032] Configure beef extract peptone nitrate solid medium with pH 4.0 at 1.05kg / cm 2 , After 20 minutes of sterilization, pour it into a sterilized petri dish under aseptic conditions, and prepare it into a flat plate for later use. Beef extract peptone nitrat...

Embodiment 2

[0041] Example 2: Detection of denitrification ability of Bacillus subtilis KD203-3 resistant to acid denitrification

[0042] In order to detect the denitrification ability of acid-resistant denitrifying Bacillus subtilis KD203-3 under different pH conditions, a single colony ring of acid-resistant denitrifying Bacillus subtilis KD203-3 strain was selected, and received respectively with 100mL sterilized pH4.0 - 250mL Erlenmeyer flasks containing nitrate-glucose denitrification liquid medium with five pH gradients of 7.5, placed at 30°C and 12Or / min for constant temperature shaking culture for 4 days.

[0043] Experiments show that the acid-resistant and denitrifying Bacillus subtilis strain KD203-3 subjected to ultraviolet mutagenesis has the lowest nitrogen removal rate of 88.35% and the highest 100% under the condition of nitrate nitrogen content of 277mg / LpH4.0-7.5. The specific results are shown in Table 1:

[0044] Table 1:

[0045]

Embodiment 3

[0046] Example 3: The present invention provides a composite microbial agent, which is made of acid-denitrification-resistant Bacillus subtilis KD203-3 as the main component, supplemented by actinomycetes and Trichoderma viride. Select ten-year-long tomato greenhouses and arrange orthogonal experiments. Each treatment is repeated 3 times. After 30 days, 20 cm of the soil surface is taken for mixing. After the soil samples are pretreated, the pH, nitrate nitrogen content, soil biomass and enzymes are measured. Liveness index determination. CK is blank control.

[0047] The test results show that the optimal volume ratio between the components is: Bacillus subtilis with the preservation number CGMCC No. 11624: actinomycetes and Trichoderma viride = 6:3:1. The three are mixed according to the above volume ratio, and the composite microbial bacterial agent of the present invention is obtained. The pH of the soil is increased to 6.16, and the removal rate of nitrate nitrogen reach...

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Abstract

The invention relates to an acid-resistant denitrifying Bacillus subtilis strain, and a composite microbial inoculant and a preparation method thereof. The acid-resistant denitrifying Bacillus subtilis strain bred by ultraviolet mutagenesis aims to cope with the high-acidity and salinized environment generated by the accumulation of the greenhouse nitrate in northern China, and has the effect of efficiently removing nitric acid nitrogen in the greenhouse soil in northern China. The invention also relates to a composite microbial inoculant and a preparation method thereof. The composite microbial inoculant is prepared by using Bacillus subtilis with the collection number of CGMCC No.11624 as the main strain and actinomycete and Trichoderma viride as auxiliary strains. The composite microbial inoculant has higher activity in greenhouse soil in northern China, has favorable restoration effects on salinization and acidification of soil in the greenhouse, and greatly enhances the bioactivity of the greenhouse soil.

Description

technical field [0001] The invention relates to an acid denitrification-resistant Bacillus subtilis and a composite microbial agent. Background technique [0002] In the past 20 years, greenhouse vegetables have developed rapidly in the northern region, but most vegetable farmers have neglected the management of soil in greenhouses during the production process. In pursuit of the goal of high-quality, high-yield, and high-efficiency vegetables in greenhouses, efforts should be made to increase the multiple cropping index and blindly increase the amount and frequency of fertilization. For example, in Shandong Shouguang vegetable greenhouse, the amount of fresh livestock and poultry manure used in the cultivation process is generally 90-150m3 / hm 2 , up to over 200m3 / hm 2 ; The amount of fertilizer input is generally 7500kg / hm 2 Or so, far beyond the absorption of vegetables. A large amount of nitrogen-containing chemical fertilizers and organic fertilizer decomposition pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/14C09K17/14C12R1/125C12R1/885C09K101/00C09K109/00
CPCC09K17/14C09K2101/00C09K2109/00C12N1/14C12N1/20C12N1/205C12R2001/125
Inventor 梁涛张海立王明英
Owner 山东黄河谷农业科技有限公司
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