Bacillus subtilis tkm-1 and its application

A Bacillus subtilis and bacteria agent technology, applied in Bacillus subtilis TKM-1 and its application fields, can solve the problems of soil, water and air pollution, easy damage to the surrounding ecological environment, increase of pesticide residues, etc., to prevent rot and lesions The effects of expansion, less drug resistance, and smaller lesions

Active Publication Date: 2021-03-02
北京泰克美高新技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the prevention and control of fruit tree rot is mainly based on chemical control, which is costly and incomplete. Long-term repeated and large-scale use of chemical pesticides will cause soil, water and air pollution, increase pesticide residues, and food safety problems are becoming more and more serious; and chemical Pesticides can easily destroy the surrounding ecological environment while killing pathogenic bacteria

Method used

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  • Bacillus subtilis tkm-1 and its application
  • Bacillus subtilis tkm-1 and its application
  • Bacillus subtilis tkm-1 and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1 Isolation and Identification of Bacillus subtilis TKM-1

[0040] 1. The bacterial strain TKM-1 that can prevent and treat apple rot disease in the present invention is collected from an apple plantation in Xiaosigou Town, Pingquan City, Hebei Province. It is isolated from the lesion sample by conventional methods during the process of isolating the pathogen of apple rot disease. of. Select tissue blocks with lesions, disinfect the surface with alcohol and saline, and wash with sterile water several times. After the surface of the tissue is dried, put the tissue bacteria solution on the surface of the PDA plate and cultivate it in a 28°C incubator. Observe once a day and record the surface changes of the plate. During the growth process, fungal hyphae and bacteria appeared at the same time, and the bacterial colonies that inhibited the fungi and formed an inhibition zone were picked for purification and culture, and activated with LB medium, and placed in a 4°...

Embodiment 2

[0052] Embodiment 2 Bacillus subtilis is measured to the inhibitory rate of rot pathogen

[0053] The inhibitory effect of Bacillus subtilis TKM-1 on the pathogenic bacteria of apple rot was detected by the plate confrontation method, and the determination method was the same as that of step 2 in Example 1. The pathogen of apple rot disease, Valsa mali, was used to screen the diseased plant samples. Through multiple parallel experiments, the inhibition rate of the strain Bacillus subtilis TKM-1 on apple rot is shown in Table 1. The zone of inhibition of Bacillus subtilis TKM-1 against pathogenic bacteria see Figure 4 .

[0054] Table 1 obtains the inhibitory rate to the pathogenic bacteria of apple rot disease through three parallel tests

[0055]

Embodiment 3

[0056] Embodiment 3 Bacillus subtilis TKM-1 aseptic filtrate measures the inhibitory rate to rot pathogenic bacteria

[0057] Preparation of fermentation filtrate: Inoculate Bacillus subtilis TKM-1 in a 250mL Erlenmeyer flask containing 100mL LB liquid medium, shake at 30°C for 24-48h to OD 600 =2.65, after the fermented liquid obtained was centrifuged at 10,000 r / min for 10 min, the supernatant was filtered with a 0.22 μm microporous membrane to obtain a sterile filtrate.

[0058] Use an Oxford cup to reserve two holes on the PDA plate. The distance between the two holes and the inoculation method of pathogenic bacteria are the same as in step 2 in Example 1. Take 100 μL of sterile filtrate and add it to the hole. For the control test, use sterile water instead of the filtrate and incubate at 28°C. After 4 days, the antibacterial effect of the filtrate was observed, and the diameter of the colony was measured to calculate the antibacterial rate. Three groups of parallel expe...

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Abstract

The present invention relates to bacillus subtilis TKM-1 and application thereof. The Bacillus subtilis TKM‑1 of the present invention has a preservation number of CGMCC NO.14950. The invention obtains a bacillus subtilis TKM-1 with antagonistic effect on the pathogenic bacteria of the rot disease through screening on the diseased strain of the apple rot disease. The fungus is made into a fungal agent or expanded in the field to prevent and control the rot disease by spraying and smearing the fruit and branches. Experiments have proved that the method can effectively prevent and treat apple rot disease, degrade the diseased rotten bark, be harmless to humans and animals, be environmentally friendly, not easy to produce drug resistance, fast wound healing, low recurrence rate of disease scars, and can It can effectively avoid a series of problems caused by excessive application of chemical pesticides, and has broad application prospects in agricultural production.

Description

technical field [0001] The invention belongs to the field of biological control, and in particular relates to bacillus subtilis TKM-1 and application thereof. Background technique [0002] According to statistics from the Food and Agriculture Organization of the United Nations (FAO), since 1988, China's apple cultivation area has occupied the first place in the world, and its cultivation area and total output account for about 2 / 5 and 1 / 3 of the world's total apple cultivation area and total output. 3. With the increase in production and successive plantings, various diseases of apples have occurred frequently, among which apple rot is one of the most widespread and serious fungal diseases. [0003] Apple rot disease is commonly known as stinky skin disease, rotten skin disease, and serial skin disease. The pathogen in East Asia is Valsa mali Miyabe et Yamada, which belongs to the subphylum Ascomycota. The ascospores are formed in autumn, and the ascospores are colorless, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/125
CPCA01N63/00A01N63/10C12N1/205C12R2001/125
Inventor 王兵于尧杜建涛高鹏鹏殷淑超李靖
Owner 北京泰克美高新技术有限公司
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