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Bacillusamyloliquefaciens strain and application thereof

A technology of amylolytic spores and bacillus, applied in the direction of application, bacteria, biocide, etc., to achieve a strong effect of inhibiting 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 no domestic patents on the use of Bacillus amyloliquefaciens to control plant fungal diseases. Internationally, only US patent 6,960,342 discloses a strain of Bacillus amyloliquefaciens isolated from the soil of Lily Field in Taiwan, which is used as a biocontrol agent. fungus to control Botrytis cinerea caused by Botrytis ellipsoides [10]
In addition, the biocontrol function of biocontrol bacteria also involves their colonization in different soil environments, but so far there is no Bacillus amyloliquefaciens biocontrol agent that can be applied to various soil types

Method used

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  • Bacillusamyloliquefaciens strain and application thereof
  • Bacillusamyloliquefaciens strain and application thereof
  • Bacillusamyloliquefaciens strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Isolation and purification of strains

[0021] From the eggplant soil in Anning District, Lanzhou City, Gansu Province, select an appropriate location, take soil samples at 5-20 mm, and isolate bacteria from it by the dilution plate method:

[0022] (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.

[0023] (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 slabs. Incubate at 30°C in an inverted incubator.

[0024] (3) Picking a single colony: After 3 to 4 days, after the colony grows on the plate...

Embodiment 2

[0029] Identification characteristics of strains

[0030] 1. Bacterial morphology observation and staining reaction

[0031] I7 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).

[0032] Staining results: I7 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 swollen cysts.

[0033] 2. Effects of environmental factors on strain I7

[0034] 2.1 O 2 Effect on strain I7

[0035] Determination of O by the deep agar method 2 For the effect on the growth of I7, insert I7 into a test tube containing glucose beef...

Embodiment 3

[0080] Preparation of cell-free culture medium of strain I7

[0081] ① Inoculate I7 ​​in 25ml small amount of NA liquid medium (pH7.0) at the 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. That is, I7 cell-free culture medium.

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Abstract

The invention relates to a strain of lys-starch bacillus as well its application. The bacterium is separated from the soil on eggplant fields, anning district, Lanzhou, Gansu. The bacterium is identified to be Bacillus amyloliquefaciens. It's preserved in the Chinese typical culture preserving center (CCTCC) with a preserving name of Bacillus amyloliquefaciens I7 and preserving code of M206027. The broth of M206027 and M206027strain, non-cell culture fluid of M206027 strain's broth, and the monomer extracted from the M206027strain, M206027 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 strain of Bacillus amyloliquefaciens, which is preserved in China Center for Type Culture Collection CCTCC, with preservation name I7 (Bacillus amyloliquefaciens) and preservation number M206027. The strain is isolated from eggplant soil in Anning District, 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 contr...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A61P3/00C12R1/07
Inventor 李红玉王亚丽支德娟
Owner LANZHOU UNIVERSITY
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