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Bacillus subtilis for preventing and treating nematode diseases and application thereof

A technology of Bacillus subtilis and Bacillus, applied in the direction of chemicals, applications, and nematocides for biological control, which can solve the problems of less biocontrol agents, etc., achieve a well-developed root system, significant control effects, and inhibit the growth of nematode larvae Effect

Active Publication Date: 2021-07-23
山东蔚蓝生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many reports about Bacillus and Pseudomonas. Among Bacillus, Bacillus subtilis and Bacillus thuringiensis are mainly used for the control of plant diseases and insect pests. Among them, Bacillus thuringiensis is the most successful bacterial insecticide developed and can be used to control A variety of insects on garden plants, especially Lepidoptera pests; Pseudomonas can multiply in the soil around plant roots to protect plants from pathogens, among which Pseudomonas fluorescens has been studied in recent decades The most reported type of biocontrol bacteria, but few can be made into biocontrol agents for wide application and mass production

Method used

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  • Bacillus subtilis for preventing and treating nematode diseases and application thereof
  • Bacillus subtilis for preventing and treating nematode diseases and application thereof
  • Bacillus subtilis for preventing and treating nematode diseases and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Isolation and screening of embodiment 1 bacterial strain

[0028] 1. Sample source: Cucumber rhizosphere soil in vegetable planting greenhouses in Jimo District, Qingdao City, Shandong Province.

[0029] 2. Strain isolation: using gradient dilution coating method

[0030] Weigh 10 g of soil sample into a 250 mL Erlenmeyer flask with sterile glass beads and 90 mL of sterile water, and shake well. Carry out gradient dilution on ultra-clean bench after standing still, obtain 10 -4 、10 -5 、10 -6 Take 100 μL of each of the three gradient dilutions and spread them on the nutrient agar plate, and repeat 3 times. Afterwards, culture them in a 37°C incubator, observe the growth after 24 hours, classify them according to the characteristics of colony size, shape and color, and pick a single colony for purification and culture. Finally, 4 strains were selected, numbered A1-A4, and kept in liquid glycerin.

[0031] 3. Screening of bacterial strains: contact killing experiment...

Embodiment 2

[0037] Identification of embodiment 2 bacterial strain ZB1

[0038] 2.1 Identification of colony morphology

[0039] Such as figure 1 As shown, the colony of strain ZB1 on the nutrient agar medium is round, raised and sticky, with neat edges, smooth surface, opaque, and the color of the colony is milky white; it can produce spores; the spores are oval, partially round or nearly round , Gram staining was positive.

[0040] 2.2 Molecular identification of 16S rRNA

[0041] The genome of strain ZB1 was extracted using a kit. Then using the genome as a template, the 16S rRNA was amplified using specific primers. The amplified PCR product was detected by 1% agarose gel electrophoresis, and the result showed that the size of the PCR product was about 1500bp, which met the requirements. The PCR amplification product was sent to a sequencing company for sequencing.

[0042]Sequencing results showed that the sequence of the PCR amplification product was SEQ ID NO:1. The sequence...

Embodiment 3

[0049] Example 3 Evaluation of Bacillus subtilis ZB1 Enzyme Production Ability

[0050] 1. Preparation of bacterial solution

[0051] Activate Bacillus subtilis ZB1, pick the activated Bacillus subtilis ZB1 and inoculate it in nutrient broth medium, cultivate it at 37°C, 220r / min for 18h, and obtain a viable count of 10 8 -10 9 CFU / ml of bacterial liquid.

[0052] 2. Preparation of crude enzyme solution:

[0053] The ZB1 strain was inoculated into the nutrient broth medium, cultured at 200r / min at 37°C for 3 days, and the fermentation broth was centrifuged at 12000r / min, and the supernatant was the crude enzyme solution.

[0054] 3. Chitinase activity assay method

[0055] Preparation of colloidal chitin: Weigh 10g of chitin powder and slowly add 500mL of concentrated hydrochloric acid, stir continuously to dissolve completely, then filter, add 4L of pre-cooled deionized water to the filtrate, overnight at 4°C, and centrifuge the next day. Precipitate, wash and centrifuge...

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Abstract

The invention relates to the technical field of functional microorganism screening and application, in particular to a novel bacillus subtilis ZB1 with the preservation number of CCCTCC (China Center for China Center for Type Culture Collection) NO: M2020741, and provides application of the novel bacillus subtilis ZB1 in agricultural production. The bacillus subtilis can effectively prevent and treat root-knot nematode diseases, also has a good prevention and treatment effect on fungal diseases such as cucumber powdery mildew and the like, and is wide in application prospect.

Description

technical field [0001] The invention relates to the technical field of functional microorganism screening and application, in particular to a bacillus subtilis for preventing and treating nematode diseases and its application in agricultural production. Background technique [0002] Crop diseases will occur during the growth process, which will have a great impact on modern agricultural production. It is imminent to prevent crop diseases. The main ways to prevent and control diseases include chemical control, physical control, biological control and so on. Since the 1980s, people have used biological control for the treatment and application of crop diseases and pests. Biological control refers to the use of one organism to fight against another organism, so as to achieve the purpose of preventing and controlling crop diseases. It has the characteristics of high efficiency, safety, and environmental friendliness, and has been paid attention to by researchers at home and abr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/22A01P5/00A01P3/00C12R1/125
CPCA01N63/22
Inventor 凌红丽吕宾苑伟伟张浩周英俊梁莉赵磊
Owner 山东蔚蓝生物科技有限公司
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