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Bacillus subtilis microbial agent and preparation method and application thereof

A technology of Bacillus subtilis and bacteria agent, applied in the field of microorganisms, can solve the problems of loss of biodiversity, plant death, increase of water and soil erosion, etc., and achieve the effect of promoting growth and reproduction and good antibacterial ability

Pending Publication Date: 2021-11-05
山东京青农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the obstacle of continuous cropping of old ginseng land has become a major technical problem that has plagued the development of ginseng industry
The growing environment of Panax notoginseng is special, the planting area is limited, and there are serious continuous cropping obstacles
Continuous cropping shows that almost all plants die, and shortening the rotation period shows severe disease and low seedling preservation rate, resulting in low yield and poor quality
[0004] At present, the easiest way to solve the obstacle of continuous cropping is to rotate crops and plant in different places. However, ginseng, American ginseng and Tianqi are perennial herbaceous plants. 2 In old ginseng areas such as Fusong, Jingyu, and Ji'an, there are basically no forests to cut down; although ginseng is planted (planted) at the same time as the forest, but due to the single tree species, the loss of biodiversity, the obvious increase of soil erosion, and the ability to conserve water resources A series of ecological problems, such as the decrease of soil fertility, the imbalance of forest ecological balance, etc., have posed a serious threat to the ecological environment of Changbai Mountain.

Method used

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  • Bacillus subtilis microbial agent and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The preparation of embodiment 1 bacterial agent

[0024] (1) Strain activation: inoculate the low-temperature preserved Bacillus subtilis JQ-0101D strain on LB solid medium, and culture at 36°C for 36h; pick a single colony and transfer it to LB slant medium, and culture at 36°C for 20h , wash the bacteria on the surface of the culture medium with sterile water as the inoculum.

[0025] (2) Preparation of first-level seed liquid: insert step into seed culture medium according to the inoculum size of 10% (volume percentage)

[0026] (1) The prepared inoculum was cultured on a shaking table for 20 h at a rotational speed of 220 rpm and a temperature of 36° C., as a primary seed solution.

[0027] 3) Secondary seed liquid preparation: according to the inoculum size of 3% (volume percentage), the step of inserting into the seed medium

[0028] (2) The first-grade seed solution was prepared with an aeration rate of 1.0vvm, a stirring speed of 220rpm, and a culture time of ...

Embodiment 2

[0033] Example 2 Antibacterial Detection

[0034] In a plate with a partition in the middle, pipette 200 μL of the prepared 10 9 CFU / ml of Bacillus subtilis fermentation broth on LB medium, spread evenly with a sterilized coating rod, and wait for its surface to dry; while inoculating three kinds of pathogenic bacteria on PDA medium respectively, place the plate at 25°C under cultivation. Do 3 parallel groups, use LB medium coated with sterile saline as a control, and observe the antibacterial effect after 72 hours.

[0035] The experimental results are shown in Table 1

[0036] Types of pathogenic bacteria

[0037] Inhibitory effect

[0038]

[0039] Note: + the inhibitory effect is weak, the inhibition rate of the colony radius is less than 10%, which affects the growth of fungal mycelia; ++ the antibacterial effect is obvious, the inhibition rate of the colony radius is between 10% and 20%; The rate is more than 20%; the average of three repetitions is taken.

[00...

Embodiment 3

[0041] Example 3 Effect on Acidobacteria (family) (qualitative experiment)

[0042] Taking Citrobacter as the representative of Agrobacterium (family), perform the following two operations respectively, and do 3 groups in parallel:

[0043] 1) Inoculate into MRS basal medium and shake directly, after 8 hours, detect OD600=0.6;

[0044] 2) Take the JQ-0101D fermentation broth, filter out the hyphae, and replace the water in the MRS basal medium. Other components remain unchanged. The improved medium is inoculated with Citrobacter at a 5% inoculation amount, and then the shaker 8h, detect OD600=0.8.

[0045] It can be seen that the fermentation product of Bacillus subtilis JQ-0101D can promote the reproduction of acid bacteria.

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Abstract

The invention provides a bacillus subtilis microbial agent, and belongs to the technical field of microorganisms. Specifically, bacillus subtilis is JQ-0101D and is preserved in the China General Microbiological Culture Collection Center, the address of the preservation unit is No. 1 Courtyard, West Beichen road, Chaoyang District, Beijing, the preservation time is March 19, 2021, and the preservation registration number is CGMCC No. 22039. The bacillus subtilis is cultured and fermented to form the microbial agent, and the microbial agent can effectively inhibit pythium, phytophthora and fusarium, has a good biocontrol effect on root rot of araliaceae ginseng plants, and has a certain promoting effect on growth and reproduction of acidobacteria.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a bacillus subtilis inoculant and its preparation method and application. Background technique [0002] Continuous cropping obstacle refers to the abnormal growth and development of crops caused by continuous cultivation of the same crop or related crops on the same soil. Symptoms are generally poor growth and development, decreased yield and quality, and in extreme cases, local dead seedlings, no seedlings or vigorous seedlings; most of the affected plants have browning, reduced branches, low vigor, and narrow distribution range, resulting in water absorption. , The capacity of nutrients is reduced. [0003] Ginseng and American ginseng are strictly forbidden to be continuously used as medicinal plants. If ginseng is continuously planted in heavy stubble land, the survival rate of ginseng seedlings will generally drop to below 30% after the second year, and the fibrous r...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/125
CPCC12N1/20A01N63/22
Inventor 郭中信耿进华尤龙王延刚
Owner 山东京青农业科技有限公司
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