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Bacillus safensis for producing protease and resisting botrytis cinerea and application thereof

A technology of bacillus and bacterial strains, which is applied in the application field of Bacillus suffolus SH180 and its protease production and resistance to tomato gray mold, which can solve the problems of few reports of bacillus, achieve the effect of inhibiting gray mold disease and promoting tomato growth

Active Publication Date: 2018-07-24
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many reports on the inhibition of Botrytis cinerea by Bacillus, but there are few reports on Bacillus that can simultaneously produce protease, resist Botrytis cinerea and significantly promote growth

Method used

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  • Bacillus safensis for producing protease and resisting botrytis cinerea and application thereof
  • Bacillus safensis for producing protease and resisting botrytis cinerea and application thereof
  • Bacillus safensis for producing protease and resisting botrytis cinerea and application thereof

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Experimental program
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Effect test

preparation example Construction

[0036] In yet another specific embodiment of the present invention, a method for preparing growth-promoting bacteria fermentation liquid is provided, the steps are as follows: transfer Bacillus safensis SH180 to a test tube with 5ml liquid medium, put Cultivate in a constant temperature shaker for 12-14h, temperature: 37°C, speed: 180-200rpm; insert the activated bacterial solution into 100-200mL medium according to the inoculum amount of 1%, put it into the shaker for 24h, temperature : 37°C, rotating speed: 180-200rpm, make fermentation broth.

[0037] In yet another specific embodiment of the present invention, a method for using the above growth-promoting bacterium fermentation liquid is provided. After the tomato seedlings are settled, the prepared fermentation liquid is diluted and then root-irrigated. In the pot test, each plant is 5 mL.

[0038] In yet another specific embodiment of the present invention, a pharmaceutical composition is provided, which is characterized...

Embodiment 1

[0040] Example 1 Determination of SH180's ability to promote plant growth

[0041] The CK strain was selected as the control, and the experiment was carried out in natural soil by planting in nutrient trays. It was cultivated in the seedling medium until 5-6 true leaves were grown, and healthy and uniform tomato seedlings were selected for use. Treat the tomato seedlings by filling the roots, the method is as follows: After transplanting the seedlings, wait for the growth of the seedlings to stabilize, and treat 5 seedlings (1 strain / pot) with the bacterial agent after 7 days, and apply 5 mL of growth-promoting bacteria fermentation liquid (concentration is about 10 8 cfu / mL), watered around the root system after appropriate dilution, and managed normally; the control was the culture solution of CK strain. After 40 days, the plant height, ground diameter and root length of the plants were measured. The comparison results are shown in Table 1 and Figure 4 shown.

[0042] Ta...

Embodiment 2

[0045] Example 2 Determination of SH180 Protein Production Ability

[0046] The determination adopts the Folin phenol method. Add 1 mL of the original fermentation broth plus 2 mL of casein solution with a mass fraction of 0.5% at a pH value of 7.0, bathe in water at 37°C for 15 minutes, then inactivate with 10% trichloroacetic acid, and then take 1 mL Add 1 mL of Folin phenol and 5 mL of 0.55 mol / L Na to the centrifuged supernatant of the above reaction liquid 2 CO 3 , in a water bath at 37°C for 15 minutes, and measure the absorbance value at 660nm. The reaction system with inactivated enzyme solution was used as the blank control. Under this reaction condition, it is defined that the amount of enzyme needed to generate 1 μg of tyrosine per 1 min is one enzyme activity. The effect of protease production is as image 3 shown.

[0047] The protease detection results are shown in Table 2 below:

[0048] Table 2 produces protease detection results

[0049]

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Abstract

The invention discloses bacillus safensis for producing protease and resisting botrytis cinerea. The bacterial strain is classified and named as Bacillus safensis SH180 and is preserved in CGMCC (China General Microbiological Culture Collection Center) on April 8, 2018; the preservation number is CGMCC15565. The bacterial strain has the effects of promoting the biological growth, producing the protease and resisting the botrytis cinerea. The bacterial strain is applied to production; important significance is realized for improving the tomato yield, reducing the harm of pesticide to irrigationwater bodies and reducing the generation of drug-resistance bacteria.

Description

technical field [0001] The invention relates to a microbial strain and an application thereof, in particular to a Bacillus safensis SH180 capable of resisting tomato gray mold and producing protease and the application thereof for producing protease and resisting tomato gray mold. Background technique [0002] As one of the main economic fruits and vegetables in our country, tomato is widely planted in our country, but with the promotion of planting, many problems have gradually become prominent. For example, fruit farmers continuously plant tomatoes in greenhouses, and the warm and humid environment in greenhouses is prone to cause fungal infections such as tomato gray mold. If chemical drugs are used, it is easy to cause too much pesticide residue on the surface of the fruit, which increases the risk of eating. In addition, during the irrigation process, pesticides dissolve in water and enter the natural environment, possibly polluting water bodies. On the other hand, ve...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P3/00A01P21/00C12R1/07
CPCA01N63/10C12N1/20C12N1/205C12R2001/07
Inventor 杜秉海丁延芹汪城墙刘凯姚良同
Owner SHANDONG AGRICULTURAL UNIVERSITY
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