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Preparation method of biocontrol bacteria fermentation broth for preventing and controlling apple tree rot diseases

A technology of apple tree rot and fermentation liquid, which is applied in the fields of botanical equipment and methods, pest control, gardening methods, etc., can solve the problems of large differences in the content of subtilisin active substances and unstable control effects, and achieve the goal of preventing and curing Effect improvement, stability improvement, permeability improvement effect

Active Publication Date: 2016-02-03
山西农业大学植物保护学院(山西省农业科学院植物保护研究所)
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AI Technical Summary

Problems solved by technology

[0004] However, the contents of active substances such as subtilisin, polymyxin, nystatin, and gramicidin produced by different sources of B. There was a significant difference in the antibacterial effect
In addition, there are many problems in the control of apple tree rot disease with the fermentation liquid of simple biocontrol bacteria, such as unstable control effect and how to effectively act on pathogenic bacteria with antagonistic bacteria and antibacterial substances.

Method used

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  • Preparation method of biocontrol bacteria fermentation broth for preventing and controlling apple tree rot diseases
  • Preparation method of biocontrol bacteria fermentation broth for preventing and controlling apple tree rot diseases
  • Preparation method of biocontrol bacteria fermentation broth for preventing and controlling apple tree rot diseases

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Screening Antagonistic Bacteria of Apple Tree Rot Pathogen

[0036] Specimens were collected in the apple orchards of Yuncheng City and Linfen City, the main apple producing areas in Shanxi Province, and 12 strains of antagonistic bacteria were isolated from the bark of apple trees. The traditional plate confrontation culture method was used for preliminary screening, and the antagonistic bacteria with better antagonistic effect were screened. . Then the obtained primary screened strains were subjected to liquid fermentation in NB medium, and the fermentation filtrate was used to carry out secondary screening, and finally obtained strains LF17 and LF18 (see Table 1) with significant antagonistic effects on the apple tree rot pathogen.

[0037] Table 1 The inhibitory effect of antagonistic bacteria fermentation broth on putrefaction bacteria

[0038]

[0039]

[0040] Identification of Antagonistic Bacteria of Apple Rot Pathogen

[0041] Morphological characteris...

Embodiment 2

[0047] Preparation of antagonistic bacteria fermentation broth

[0048] Activation of the antagonistic bacteria obtained by screening: first inoculate the preserved strains LF17 and LF18 on NA medium, and incubate at a constant temperature of 25°C for 2 days.

[0049] Pick the activated strains LF17 and LF18 and inoculate them in NB medium with a bottled volume of 80mL / 250mL; 150r min -1 , 25°C constant temperature shaking culture for 5 days to obtain Bacillus subtilis fermentation broth.

[0050] Filtration of the fermented broth: filter the fermented broth prepared in the previous step with filter paper to obtain the stock solution of the fermented broth.

[0051] Concentrated fermented broth

[0052] The fermented broth prepared in the previous step was collected and concentrated, concentrated to 1.5 times the original fermented broth concentration and 3.0 times the original fermented broth concentration, and stored at 4°C for future use.

Embodiment 3

[0054] Test of Antagonistic Bacteria Fermentation Broth Inhibiting Apple Tree Rot Bacteria

[0055] Azoxystrobin, octoprobin, difenoconazole, and thiophanate-methyl were prepared into a 1000 μg / mL stock solution with acetone, and the stock solution was stored in a 4°C refrigerator for later use. When used, dilute them with sterile water to form a series of concentrations. Under a sterile environment, mix the prepared series of concentrations with the PDA medium cooled to about 40°C in a ratio of 1:9 and shake well to make a PDA plate containing 4 kinds of original drugs.

[0056] Concentrate the prepared fermentation filtrate to 1.5 times the concentration of the original fermentation broth and 3.0 times the concentration of the original fermentation broth, add 8.5g / L agar powder, mix and shake well, and sterilize by high pressure steam at 121°C for 20min. Finally, a plate containing the fermentation broth was made.

[0057] The preserved strains were inoculated into PDA med...

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Abstract

The present invention discloses a preparation method of a biocontrol bacteria fermentation broth for preventing and controlling apple tree rot diseases. According to the present invention, two strains such as LF17 and LF18 having a good apple tree rot disease bacteria inhibition effect are separated and screened from the fruit tree branches in the Shanxi Province apple production area Linfen City apple orchard, and the two strains are identified as B.subtilis and through strain colony characteristic analysis, strain microscopic characteristic analysis and 16SrDNA sequence analysis; the biocontrol bacteria fermentation broth preparation steps comprise: S1, culture medium preparing, S2, antagonistic strain activation, S3, antagonistic strain liquid culture, S4, fermentation forth filtration and concentration, S5, antagonistic bacteria fermentation broth field prevention and control agent preparation, and S6, antagonistic bacteria fermentation broth paste field prevention and control agent preparation; and the obtained biocontrol bacteria fermentation forth has characteristics of effective apple rot disease prevention and control, relative safety to humans and animals, good environmental compatibility, drug resistance generation resistance, rapid wound healing, and low scar recurrence rate.

Description

technical field [0001] The invention relates to the field of prevention and treatment of apple tree rot, in particular to a method for preparing a fermentation liquid of biocontrol bacteria for preventing and treating apple tree rot. Background technique [0002] Apple tree rot is one of the most serious diseases in my country's apple production areas. The disease causes apple tree bark to rot, tree vigor to weaken, and apple yield and quality to decline; when it is severe, it causes the main stem to die, and even causes the garden to be destroyed. At present, rot disease seriously restricts the development of apple industry. The survey of the modern apple industry technology system by the Ministry of Agriculture shows that the overall incidence of apple tree rot disease is 52.7%. With the increase of tree age, the incidence rate of rot disease increases, the incidence rate of fruit trees aged 4-10 is 26.8%, the incidence rate of fruit trees aged 11-17 is 54.0%, and the in...

Claims

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Application Information

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IPC IPC(8): A01N63/02A01P3/00A01P21/00A01G7/06A01G17/00A01N63/00A01N43/90A01N43/12
Inventor 赵晓军殷辉周建波张志斌姚众秦楠
Owner 山西农业大学植物保护学院(山西省农业科学院植物保护研究所)
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