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Bacillus subtilis DYr3.3 with broad-spectrum bacteriostatic activity and preparing method and application

A technology of bacillus subtilis and bacteriostatic activity is applied in the field of plant disease biological control, and can solve the problems of not meeting application needs, unsatisfactory strain effect, low working concentration and the like

Active Publication Date: 2018-06-12
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above studies mainly use various bacillus to control postharvest apple ring spot and anthracnose, but the effect of each strain is not ideal, and the working concentration is low (stock solution or 20 times slightly diluted), which cannot meet the actual application needs in production
Some people also screened and used Bacillus subtilis to prevent and control postharvest pear ring spot, and isolated 21 strains of Bacillus subtilis from pear samples, which can inhibit the rot of pear fruit after inoculation with Pear ring spot, and the efficiency of inhibiting the expansion of lesions is more than 40%. [Ding Fangbing, Screening and using Bacillus subtilis to control pear ring spot, Journal of Jiangsu Agricultural Science, 2009, 25 (5): 1002-1006], still far from meeting the needs of practical application in production

Method used

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  • Bacillus subtilis DYr3.3 with broad-spectrum bacteriostatic activity and preparing method and application
  • Bacillus subtilis DYr3.3 with broad-spectrum bacteriostatic activity and preparing method and application
  • Bacillus subtilis DYr3.3 with broad-spectrum bacteriostatic activity and preparing method and application

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Embodiment 1

[0033] A preparation method of Bacillus subtilis with broad-spectrum antibacterial activity, the steps are:

[0034] (1) Collect the roots of Zizhi rose in Daye Rose Field, Huangshi City, Hubei Province, cut the roots into tissues about 0.5 cm in size, disinfect with 70% (volume ratio) alcohol for 1 min, and rinse with sterile water 2-3 times After that, place it on PDA medium for dark culture at 25°C. After 36 hours of culture, transfer the grown bacterial colony to a new PDA medium to separate and purify a single colony by streaking;

[0035] The preparation method of described PDA culture medium is: 200 g potato, 20 g sucrose, 15 g agar powder are dissolved in 1000 mL distilled water, adjust pH value to about 7.0;

[0036] (2) After separation and purification, an antagonistic bacterium Bacillus subtilis ( Bacillus subtilis ) strain DYr3.3, the nucleotide sequence of which is shown in SEQ ID NO: 1, the strain DYr3.3 was deposited in the China Center for Type Culture Coll...

Embodiment 2

[0055] A broad-spectrum antibacterial activity of Bacillus subtilis ( Bacillus subtilis ) in the preparation or prevention and treatment of pear fruit rot medicine, the steps are:

[0056] A. Pick a single colony of DYr3.3 strain in a sterile environment and inoculate it in a container containing 90mL of the most suitable culture solution A (3.0% (mass volume ratio) glucose, 0.5% (mass volume ratio) yeast extract powder, 0.3% (mass volume ratio) sodium chloride, 0.1% (mass volume ratio) magnesium sulfate, adjust the pH value to 6.0-6.5) in a 250mL Erlenmeyer flask, 28°C, 150r / min shaking culture for 24 hours, to obtain the seed culture liquid.

[0057] B. Prepare 10 500mL Erlenmeyer flasks, each bottle contains 200 mL fermentation liquid, each bottle is added with seed culture liquid in a ratio of 1:100 (volume ratio), placed in a large-amplitude constant temperature freezer shaker, 150 r at 28 ° C / min Shaking culture for 48 h, to obtain DYr3.3 fermentation broth, the conce...

Embodiment 3

[0060] Antagonistic Antibacterial Control of Pear Fruit Rot

[0061] Dilute the Bacillus subtilis strain DYr3.3 100 times to obtain a concentration of 4.89×10 6 Bacterial suspension, and then buy fully ripe pear fruit (Hebei Crown Pear, Hebei Crown Pear, Pyrus bretschneideri Rehd . var. Huangguanli) were soaked in the prepared bacterial suspension for about 30 s, so that the surface was fully and evenly coated with bacteria, and then taken out to dry. There were 6 replicates in each treatment group, and the activated fungus cake (5 mm in diameter) was inoculated equidistantly from the waist of the fruit, and then the wet cotton sheet soaked in sterile water was covered on the inoculation site. Take another 2 pears that have not been soaked in the bacterial suspension as CK + control group. The treated pears were placed in plastic trays covered with a layer of moist gauze and plastic wrap, and the relative humidity in the trays was kept high at all times. Finally, the plast...

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Abstract

The invention discloses bacillus subtilis DYr3.3 with broad-spectrum bacteriostatic activity and a preparing method and application. The bacteriostatic bacillus subtilis strain DYr3.3 with the preservation number of CCTCC M 2018024 is prepared through the steps of firstly, collecting purple-branch rose roots, cutting the roots into tissue, disinfecting the tissue with 70% alcohol, flushing the tissue with sterile water, placing the tissue in a PDA culture medium for dark culture, transferring a growing bacterial colony onto a new PDA culture medium after culturing, and drawing a line to separate and purify a single colony; secondly, obtaining the bacteriostatic bacillus subtilis strain DYr3.3 after separating and purifying, wherein the nucleotide sequence of the bacteriostatic bacillus subtilis strain DYr3.3 is shown as SEQ ID NO:1. The strain DYr3.3 is applied to the preparation of pear fruit rot treatment or prevention medicines. The DYr3.3 bacteria solution obtained from the strainDYr3.3 through liquid fermentation can prevent pear fruit rot caused by pear ring spot and anthracnose dangers and has a good disease preventing effect on pear fruit rot, the pear fruit rot is remarkably restrained, the prevention rate can reach 100%, and the bacillus subtilis DYr3.3 is widely used for preventing and treating the pear fruit rot disease.

Description

technical field [0001] The invention belongs to the technical field of biological control of plant diseases, more specifically relates to a broad-spectrum antibacterial activity of Bacillus subtilis DYr3.3, and also relates to a preparation method of a broad-spectrum antibacterial activity of Bacillus subtilis DYr3.3, and The invention relates to the use of Bacillus subtilis DYr3.3 with broad-spectrum antibacterial activity. Background technique [0002] Since the production places of fruit and vegetable products are far away from cities, and the maturity period is relatively concentrated, a certain period of storage and transportation is required. During this period, a large amount of postharvest losses, especially the rot of fruits and vegetables, will occur. According to reports, 10%-30% of fresh fruit in developed countries is lost due to post-harvest rot, and in developing countries that lack storage and transportation refrigeration equipment, the rot rate is as high as...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/125
CPCA01N63/10C12N1/205C12R2001/125
Inventor 徐文兴荚恒霞蔡丽王利平洪霓王国平
Owner HUAZHONG AGRI UNIV
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