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Bacillus subtilis strain and application thereof

A Bacillus subtilis, strain technology, applied in the application, bacteria, fungicides and other directions, can solve problems such as narrow control spectrum, and achieve the effect of strong inhibitory activity

Inactive Publication Date: 2007-04-25
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many domestic and international literature reports and patents on the use of Bacillus subtilis to control plant fungal diseases, but on the whole, there is mainly the problem of narrow control spectrum, usually only 1 to 2 pathogenic fungi can be controlled
In addition, the biocontrol function of biocontrol bacteria also involves its colonization in different soil environments, but so far no Bacillus subtilis biocontrol bacteria can be applied to a variety of soil types, and no cell-free bacteria have been found. It is reported that the culture medium and its extract can control more than 10 strains of pathogenic fungi, usually only 1 to 2 strains of pathogenic fungi can be controlled

Method used

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  • Bacillus subtilis strain and application thereof
  • Bacillus subtilis strain and application thereof
  • Bacillus subtilis strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Isolation and purification of strains

[0020] From the soil of the lily field in Fanjiaping, Lanzhou City, Gansu Province, select an appropriate location, take a soil sample at a distance of 5-20 mm, and isolate bacteria from it by the dilution plate method:

[0021] (1) Preparation of soil dilution: weigh 10 g of soil sample, put it into 100 ml of sterile water, and oscillate on a shaker for 30 min. Set aside to clarify. Aseptically draw 100 μl of the supernatant and add it to 900 μl sterile water, dilute to 10 with sterile water -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 A total of eight concentration gradients.

[0022] (2) Cultivation: add sterilized NA (beef extract 3g, peptone 5g, glucose 10g, yeast extract 1g, agar 15g, distilled water 1000ml, pH 7.0) agar medium cooled to 55~60℃ for each dilution concentration, Made into a flat plate. Incubate at 30°C in an inverted incubator.

[0023] (3) Picking a single colony: After 3 to 4 days, after t...

Embodiment 2

[0028] Identification characteristics of strains

[0029] 1. Bacterial morphology observation and staining reaction

[0030] M22BH3 was inoculated in NA liquid medium and cultured with shaking at constant temperature for a suitable period of time, and then stained for microscopic examination. Including Gram staining (crystal shoxenamide staining method), flagella staining (silver nitrate staining method), capsule staining (wet ink method), spore staining (modified Schaeffer-Fulton's staining method).

[0031] Staining results: M22BH3 is rod-shaped bacteria, G + Sex; flagella grow laterally, with 8 to 20 flagella growing on each cell, the length of which is about 2 to 3 times that of the cell; capsulated: spore-forming, mesozoic, and spore cysts swell.

[0032] 2. Effects of environmental factors on strain M22BH3

[0033] 2.1 O 2 Effect on strain M22BH3

[0034] Determination of O by the deep agar method 2 For the effect on the growth of M22BH3, insert M22BH3 into a test ...

Embodiment 3

[0080] Preparation of cell-free culture medium of M22BH3 strain

[0081] ① Inoculate M22BH3 in 25ml small amount of NA liquid medium (pH7.0) at a ratio of 1:100, 120rpm / min, 30°C, shake culture for 24h;

[0082] ② Then inoculate the above-mentioned culture solution into 1000ml large amount of NA liquid medium (pH7.0) at a ratio of 1:20, culture at 120rpm / min, 30°C, and shake for 72h;

[0083] ③ Centrifuge the fermented liquid at 8000-12000r / min to get the supernatant, and the collected supernatant is filtered through a bacterial filter (0.22μm) to obtain a concentrated solution, which is spread on a flat plate, and no viable bacteria are detected. It is the M22BH3 strain preparation.

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Abstract

The invention relates to a strain of Bacillus subtilis as well its application. The bacterium is separated from the soil on lily fields, fanjiaping, Lanzhou, Gansu. The bacterium is identified to be Bacillus-subtilis. It's preserved in the Chinese typical culture preserving center (CCTCC) with a preserving name of Bacillus-subtilis M22BH3 and preserving code of M206028. The broth of M22BH3 and M22BH3 strain, non-cell culture fluid of M22BH3 strain's broth, and the monomer extracted from the M22BH3 strain, M22BH3 broth or its non-cell culture fluid can be used in the biological control of peach anthracnose bacterium, pear nigrities artis bacterium, apple brown rot bacterium, Fusarium acuminatum, Fusarium oxysporum, Fusarium semitectum, chilli black rot bacterium, cucumber blight bacterium, tomato early blight bacterium, eggplant early blight bacterium, dry thread Pyrenomycetes, wheat foot rot bacterium, potato thread Pyrenomycetes, potato early blight bacterium, potato late blight bacterium and watermelon downy mildew bacterium. It's easy to get the material of the invention and easy to operate; besides, it can be widely used in full scale production.

Description

technical field [0001] The invention relates to a newly screened Bacillus subtilis, which is preserved in China Center for Type Culture Collection CCTCC, with preservation name Bacillus-subtilis M22BH3 and preservation number M206028. The strain is isolated from the soil of the lily field in Fanjiaping, Lanzhou City, Gansu Province. It has a significant inhibitory effect on the growth of pathogenic fungi that can cause plant diseases, is non-toxic to humans and animals, and is friendly to the ecological environment. It can be prepared into biological pesticide products. Background technique [0002] In 1921, Hartely used antagonistic fungi to control cotton seedling damping-off (Pythium debaryanum), which was the beginning of people using antagonistic microorganisms to control plant diseases [1] . Subsequently, countries around the world have conducted extensive research on this emerging field, trying to screen out antagonistic microorganisms for the control of plant diseas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/125
Inventor 李红玉王亚丽支德娟
Owner LANZHOU UNIVERSITY
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