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Bacillus subtilis pro6a5, its bacterial agent and preparation method and application in melon cultivation

A technology of Bacillus subtilis and melon, which is applied in the field of biological control, can solve the problems of narrow genetic background of germplasm resources, destruction of ecological balance, and difficulty in popularization and implementation, and achieve the effects of improving the ecological environment, efficient prevention and control, and overcoming the obstacles of saline-alkali land stress

Active Publication Date: 2022-03-25
河南省农业科学院园艺研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, limited by factors such as the narrow genetic background of germplasm resources and limited disease-resistant materials; the most commonly used, fastest and most direct way of prevention and control is chemical drug control. Chemical drug control can reduce the occurrence of diseases to a certain extent, but due to excessive, Pesticide residues, food safety, environmental pollution, ecological balance disruption, and pathogen resistance caused by the frequent use of chemical fungicides limit the use of chemical pesticides; agricultural control and physical control can only be alleviated to a certain extent. Occurrence of disease
[0005] Cropping crop rotation is also an effective way to prevent or reduce pests and diseases, but it is difficult to popularize and implement in the case of limited per capita arable land in my country and uneven geographical distribution.

Method used

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  • Bacillus subtilis pro6a5, its bacterial agent and preparation method and application in melon cultivation
  • Bacillus subtilis pro6a5, its bacterial agent and preparation method and application in melon cultivation
  • Bacillus subtilis pro6a5, its bacterial agent and preparation method and application in melon cultivation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Isolation and screening of Bacillus subtilis

[0028] (1) Isolation and purification of soil Bacillus

[0029] Collection of melon fields (collected by Liang Shen in Duzhai Village, Putaijia Township, Lankao County on September 20, 2016) 300g of moist soil in a 5-20cm soil layer, weighed 10g after crushing, and placed in a pot filled with 100ml of sterile water. In the Erlenmeyer flask, shake for 3 to 5 minutes, and heat in a water bath at 80°C for 10 minutes. Measure the soil suspension and dilute with sterile water to prepare 10 -2 , 10 -3 , 10 -4 and 10 -5 Soil suspension; suck up the above 10 -3 , 10 -4 , 10 -5 50 μl of soil suspension was spread on NA medium (beef extract 3g, yeast powder 1g, peptone 5g, glucose 10g, agar 12g, water 1L, pH 6.8~7.0, sterilized at 121℃ for 20min) plate, in ultra-clean Plates were coated on a bench and allowed to dry. Inverted in a biochemical incubator and incubated at 37 °C for 48 h. After the single spore colon...

Embodiment 2

[0042] Example 2: Identification of Bacillus subtilis Pro6A5

[0043] (1) Analysis of physical and chemical properties

[0044] 1) Colony characteristics and bacterial morphology observation

[0045] Bacillus Pro6A5 was inoculated on NA medium and cultured in a biochemical incubator at 30 °C for 48 h. Observe the colony morphology, color and growth of the strain on the medium. Bacillus colonies cultured for 48 h were picked and Gram-stained on glass slides to observe the size and morphology of the bacteria.

[0046] 2) Determination of strain physiology and biochemical tests

[0047] Physiological and biochemical experiments of Pro6A5 strains were determined using the culture medium and experimental methods recommended in the Manual for the Identification of Common Bacterial Systems and Bacillus. Physiological and biochemical tests mainly include: methyl red reaction, contact enzyme test, starch hydrolysis test, hydrogen peroxide test, gelatin liquefaction test, casein dec...

Embodiment 3

[0062] Example 3: Preparation of microbial inoculum

[0063] The preparation steps of Pro6A5 inoculum are as follows:

[0064] (1) Strain culture and preparation

[0065] Bacillus subtilis Pro6A5 was inoculated into a test tube slanted soybean casein solid medium (tryptone 17g, soybean peptone 3g, NaCl 5g, KH 2 PO 4 2.5g, glucose 2.5g, agar powder 12g, add water to volume 1000ml, pH 7.0, sterilize at 121°C for 20min), and cultivate at 28-35°C for 24-36 hours to obtain bacterial cells;

[0066] (2) Preparation of seed solution

[0067] The test tube strain prepared in step (1) was inserted into soybean casein liquid medium (tryptone 17g, soybean peptone 3g, NaCl 5g, KH 2 PO 4 2.5g, glucose 2.5g, add water to volume 1000ml, pH 7.0, sterilize at 121°C for 20min), shake and culture at 28-35°C for 12-16 hours to make seed solution;

[0068] (3) Preparation of fermentation medium

[0069] Glucose, corn flour, soybean meal powder, dipotassium hydrogen phosphate and water were ...

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Abstract

The invention discloses a bacillus subtilis Pro6A5, its bacterial agent, a preparation method and application in melon cultivation, aiming at solving the technical problem of limited disease prevention and control in melon cultivation. In the present invention, a strain of Bacillus subtilis Pro6A5 was screened out for the first time, and it was preserved in the China Center for Type Culture Collection with the preservation number CCTCC NO.M 2017196. The microbial agent prepared by Bacillus subtilis Pro6A5 has a good effect on the control of bacterial diseases, and has a strong inhibitory effect on melon bacterial fruit spot, peach bacterial perforator, tobacco bacterial wilt, cabbage soft rot and tomato canker. ; It can also overcome the obstacle of continuous cropping of muskmelon in saline-alkali land; it is highly effective in prevention and control, easy to use, and does not cause environmental pollution.

Description

technical field [0001] The invention relates to the technical field of biological control, in particular to a kind of Bacillus subtilis ( Bacillus subtilis ) Pro6A5, its bacterial agent and preparation method and application in muskmelon cultivation. Background technique [0002] Common diseases of melon include fusarium wilt, powdery mildew, downy mildew, sclerotinia, angular spot, anthracnose, etc. The continuous cropping of melons aggravates the occurrence of such diseases and continuous cropping obstacles. , will seriously affect the yield and quality of melon. [0003] In the prior art, various measures can be taken to prevent and control diseases, such as plant quarantine, utilization of disease-resistant varieties, agricultural control, chemical control, physical control, and biological control. [0004] Disease-resistant varieties are the most simple, economical and effective prevention and control methods. Strengthen seed quarantine and do a good job in seed-car...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P1/00C05G3/60C12R1/125
CPCC12N1/20A01N63/22C05G3/60C12R2001/125C12N1/205
Inventor 赵卫星李晓慧梁慎常高正康利允高宁宁徐小利王龙飞杨作运
Owner 河南省农业科学院园艺研究所
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