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Bacillus subtilis Pro1A2, microbial agent thereof, preparation method and application of microbial agent in melon cultivation

A technology of Bacillus subtilis and melon, applied in the field of biological control, can solve the problems of uneven distribution, difficult to popularize and implement, etc., and achieve the effects of improving the ecological environment, good effect, and efficient control

Active Publication Date: 2020-10-16
兰考县搏洋农业发展有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 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 and uneven geographical distribution in my country.

Method used

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  • Bacillus subtilis Pro1A2, microbial agent thereof, preparation method and application of microbial agent in melon cultivation
  • Bacillus subtilis Pro1A2, microbial agent thereof, preparation method and application of microbial agent in melon cultivation
  • Bacillus subtilis Pro1A2, microbial agent thereof, preparation method and application of microbial agent in melon cultivation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Isolation and screening of Bacillus subtilis

[0030] (1) Isolation and purification of Agrobacterium

[0031] Collected (collected by Liang Shen in Dingqidang Village, Sanyizhai Township, Lankao County on September 20, 2016) 300g of moist tillage layer soil 5-20cm away from the ground surface, weighed 10g after crushing, and placed in a container of 100ml Shake for 3-5 minutes, then heat in a water bath at 80°C for 10 minutes. Measure the upper soil suspension and dilute with sterile water to prepare 10 -2 、10 -3 、10 -4 and 10 -5 Soil suspension; pipette the above 10 -3 、10 -4 、10 -5 Spread 50 μl of soil suspension on the plate of NA medium (3g of beef extract, 1g of yeast powder, 5g of peptone, 10g of glucose, 12g of agar, 1L of water, pH6.8~7.0, sterilized at 121℃ for 20min) Coat slabs on bench and allow to dry. Place them upside down in a biochemical incubator and incubate at 37 °C for 48 h. After monospore colonies grow, single colonies are ...

Embodiment 2

[0052] Example 2: Identification of Bacillus subtilis Pro1A2

[0053] (1) Analysis of physical and chemical properties

[0054] 1) Colony characteristics and bacterial morphology observation

[0055] Bacillus Pro1A2 was inoculated on NA medium and cultured in a biochemical incubator at 37 °C for 48 h. Observe the colony morphology and growth of the strains on the culture medium. Bacillus colonies cultured for 48 h were picked and stained with Gram on glass slides to observe the size and shape of the bacteria.

[0056] 2) Determination of Physiological and Biochemical Tests of the Strains For the determination of the physiological and biochemical tests of the Pro1A2 strain, the medium and experimental methods recommended in the "Common Bacterial System Identification Manual" and "Bacillus" were used. Physiological and biochemical tests mainly include: methyl red reaction, contact enzyme test, starch hydrolysis test, hydrogen peroxide test, gelatin liquefaction test, casein d...

Embodiment 3

[0071] Embodiment 3: the preparation of microbial bacterial agent

[0072] The preparation steps of Pro1A2 bacterial agent are as follows:

[0073] (1) Strain cultivation and preparation

[0074] Inoculate Bacillus subtilis Pro1A2 into test tube slant soybean casein solid medium (tryptone 15g, soybean protease hydrolyzate 5g, sodium chloride 5g, glucose 2.5g, agar powder 12g, add water to 1000ml, pH 7.0~ 7.5, after aliquoting, sterilize at 121°C for 20 minutes), culture at 28-35°C for 24-36 hours to obtain bacteria;

[0075] (2) Preparation of seed solution

[0076] Put the test tube bacteria prepared in step (1) into soybean casein liquid medium (tryptone 15g, soybean protease hydrolyzate 5g, sodium chloride 5g, glucose 2.5g, add water to 1000ml, pH 7.0-7.5 , after aliquoting, sterilize at 121°C for 20 minutes), shake and culture at 28-35°C for 12-16 hours to make seed liquid;

[0077] (3) Preparation of fermentation medium

[0078] Glucose, corn flour, soybean cake powd...

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Abstract

The invention discloses bacillus subtilis Pro1A2, a microbial agent thereof, a preparation method and application of the microbial agent in melon cultivation, and aims to solve the technical problemsthat the disease control effect is poor and continuous cropping obstacles are difficult to overcome in melon cultivation. The bacillus subtilis Pro1A2 is screened for the first time and preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: M 2017194. The microbial agent prepared from the bacillus subtilis Pro1A2 can have a good control effect on plantfungal diseases such as melon gummy stem blight, melon fusarium wilt, wheat take-all and rice blast, and also has a good effect on controlling bacterial diseases, such as melon bacterial fruit blotchbacteria, peach bacterial perforating bacteria, tobacco bacterial wilt bacteria, Chinese cabbage soft rot bacteria and tomato canker bacteria; the microbial agent can also overcome the obstacles of premature senility and continuous cropping of melons; the control is efficient, the use is simple, and NO environmental pollution is caused.

Description

technical field [0001] The invention relates to the technical field of biological control, in particular to a bacillus subtilis ( Bacillus subtilis ) Pro1A2, its bacterial agent and preparation method and its application in melon cultivation. Background technique [0002] Common diseases of melons include fruit spot, vine blight, fusarium wilt, powdery mildew, downy mildew, sclerotinia, angular leaf spot, anthracnose, etc. Continuous cropping in continuous cropping will aggravate the occurrence of such diseases and continuous cropping obstacles , if effective control is not carried out in time, it 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, crop rotation, agricultural control, chemical control, physical control and biological control. [0004] Disease-resistant varieties are the most convenient, economic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/22A01P1/00A01P3/00A01P21/00C12R1/125
CPCC12N1/20A01N63/22C12R2001/125C12N1/205
Inventor 梁慎赵卫星李晓慧常高正康利允高宁宁徐小利王龙飞杨作运
Owner 兰考县搏洋农业发展有限公司
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