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Fermentation process of Bacillus pumilus LD-b1 and its application in control of plant diseases

A technology for Bacillus pumilus and plant diseases, which can be used in applications, plant growth regulators, botanical equipment and methods, etc., and can solve problems such as difficulties in colonization

Inactive Publication Date: 2013-03-13
TIANJIN AGRICULTURE COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is difficult for plant disease biocontrol bacteria isolated and screened from soil and plant rhizosphere to colonize in plants, and it is difficult to gain an advantage in competition with disease bacteria

Method used

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  • Fermentation process of Bacillus pumilus LD-b1 and its application in control of plant diseases
  • Fermentation process of Bacillus pumilus LD-b1 and its application in control of plant diseases
  • Fermentation process of Bacillus pumilus LD-b1 and its application in control of plant diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Isolation of Bacillus pumilus LD-b1

[0021] Take a section of the stem of fresh clouded gentian, rinse twice with sterile water, absorb the water with sterile filter paper; soak in 70% ethanol solution for 2 minutes, then soak with 0.1% mercury for 30 seconds, rinse with sterile water 3 times . Under aseptic conditions, peel off the outer skin with a blade and cut it into small sections of about 1 cm, place them on the surface of the screening medium, and incubate at 30°C for 4-8 days. After the bacteria grow on the plate, the plate is streaked and purified to obtain a pure culture of isolated bacterial strains.

Embodiment 2

[0023] Morphological characteristics of strain LD-b1

[0024] LD-b1 was cultured on a solid medium plate at 30°C for 1 day to form a milky white opaque colony. The colony was approximately round, with irregular edges, small protrusions in the middle, 0.6-1.1cm in diameter, and no pigment secretion; bacteria of strain LD-b1 The somatic cells are short rod-shaped, with a size of 0.5μ.m~0.6μm×1.5μm~2.1μm, Gram-positive, with spores.

[0025] Physiological and biochemical characteristics of strain LD-b1

[0026] The growth temperature range of strain LD-b1 is 8~50℃, pH range is 5.5~8.5, NaCl concentration range is 0~7%; the available carbon sources are: glucose, sucrose, lactose, aldose, xylose, galactose , Mannitol, N-acetyl-glucosamine, gluconate; can not use starch, dextrin, melibiose, rhamnose and sorbitol; strain LD-b1 contact enzyme, citrate utilization, casein Hydrolysis, esculin hydrolysis, VP is positive, oxidase, urease, H 2 S production, indole production, nitrate reduction,...

Embodiment 3

[0030] Fermentation production process of strain LD-b1

[0031] Slant medium (g / L): sucrose 10, peptone 2, beef extract 3, yeast extract 0.5, agar 15, pH 7.5, sterilized at 121°C for 30 min.

[0032] Seed medium (g / L): sucrose 15, peptone 5, yeast powder 0.2, dipotassium hydrogen phosphate 0.5, magnesium sulfate 0.05, sodium chloride 0.1, pH 7.5, sterilized at 121°C for 30 min.

[0033] Fermentation medium (g / L): sucrose 25, soybean meal 3, ammonium sulfate 2.5, yeast powder 0.2, diammonium phosphate 0.8, magnesium sulfate 0.05, ferrous sulfate 0.02, sodium chloride 0.1, pH 7.5, 121°C Sterilize for 30 minutes.

[0034] Streak the conserved strains and inoculate them on the solid slant medium, incubate at 30°C for 36 to 48 hours, wash the activated strains from the slant of the test tube with 5mL sterile water, and insert them into the seed culture medium. Liquid seed culture medium, the inoculated Erlenmeyer flask is placed on a rotary shaker, and cultured at 150r / min at 30°C for 24-...

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Abstract

Belonging to the technical field of biological pesticides, the invention provides a Gentiana nubigena endophyte applicable to biological control of plant diseases and its fermentation process. The strain involved in the invention is Bacillus pumilus LD-b1, which is preserved in the General microbiological center of China Committee for Culture Collection of Microorganisms on May 14, 2012, and has a preservation number of CGMCC No.6112. Mixed strain agar plate punching method experiments show that the strain LD-b1 can inhibit the growth of a plurality of plant disease fungi. LD-b1 can substantially inhibit Valsa mali from eroding apple fruits. In greenhouse pot experiments, the LD-b1 can achieve a cucumber Mycosphaerella arachidicola control effect of 81.2%. The fermentation process consists of: culturing LD-b1 in a seed tank for 24h to a logarithmic growth phase, inoculating 5% of the LD-b1 into a production tank according to an inoculation amount, controlling the initial pH at 7.5 and the fermentation temperature at 30DEG C, maintaining dissolved oxygen over 5%, and performing fed-batch fermentation for 68h, thus obtaining a bacterial amount over 10<10>cfu / mL. With the advantages of fast growth and wide antibacterial spectrum, the strain LD-b1 has good application prospects in biological control of plant diseases.

Description

Technical field [0001] The invention relates to a biological control strain for preventing and treating plant diseases, including its fermentation production process and its application in biological control of plant diseases, and belongs to the technical field of biological pesticides. Background technique [0002] Since Vogl isolated the first endophyte from ryegrass seeds in 1898, the research on endophytes has a history of more than 100 years. Research in recent years has shown that almost all plants have endophytes, which grow in the tissues under the epidermis of plants, including bacteria, fungi and actinomycetes. According to estimates by Dreyfuss and Chapela, there are at least hundreds of thousands of endophytes that live in the special environment within or between the cells of vascular plants. my country's plant resources are extremely rich, but due to many reasons such as land reclamation, grazing, afforestation, and mining in recent years, many wild plant resources...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P3/00C12R1/07
Inventor 黄海东杨红澎李晓雁
Owner TIANJIN AGRICULTURE COLLEGE
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