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Bacillus subtilis K13, and culture method and application thereof

A technology of Bacillus subtilis and a culture method, applied in the field of Bacillus subtilis K13 and its culture, can solve the problem that the huge role of disease resistance gene resources has not been fully exerted, the situation of disease resistance rubber germplasm resources is unclear, and monitoring and early warning work technology Insufficient support and other problems, to achieve the effect of good development and application prospects, fast reproduction, and easy industrial production

Active Publication Date: 2016-09-14
ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the occurrence of diseases and changes in disaster conditions, the lack of systematic research on disease epidemics, occurrence and epidemic patterns, and weak basic research work; insufficient technical support for disease monitoring, monitoring and early warning work; lack of awareness and technical reserves for new occurrences and catastrophic dangerous diseases; The chemical control method is single, and there are ecological and drug resistance safety hazards; the situation of disease-resistant rubber germplasm resources is unclear, and the huge role of the mining and utilization of disease-resistant gene resources in disease prevention and control has not been fully utilized

Method used

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  • Bacillus subtilis K13, and culture method and application thereof
  • Bacillus subtilis K13, and culture method and application thereof
  • Bacillus subtilis K13, and culture method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Embodiment 1 Bacillus subtilis (Bacillus subtilis) K13

[0040]1. Sampling: Pick the leaves with anthracnose spots from the rubber tree, wash off the dust and gravel on the leaves with water, dry them, and use scissors to cut small diseased leaves at the junction of disease and health for later use; cut the size of the small diseased leaves It is 5mm×5mm.

[0041] 2. Sample disinfection: the small diseased leaves were put into a beaker, soaked with mercuric chloride for 30 seconds, stirred gently constantly, then rinsed three times with sterile water, and dried to obtain the small diseased leaves after disinfection.

[0042] 3. Isolation and cultivation of Bacillus subtilis (Bacillus subtilis) K13: put the sterilized small diseased leaves on the potato dextrose agar solid (PDA) medium with sterilized tweezers, place 4 pieces evenly in each dish, and then put Seal the petri dish with plastic wrap, put it upside down in a constant temperature incubator, cultivate it at 2...

Embodiment 2

[0053] Physiological and biochemical identification of embodiment 2 Bacillus subtilis (Bacillus subtilis) K13

[0054] 1. V-P test

[0055] 1) Principle

[0056] In the process of sugar metabolism, some bacteria decompose glucose to produce pyruvate, and decarboxylate pyruvate to produce acetylmethylmethanol. Acetylmethylmethanol is oxidized to diacetyl by oxygen in the air in an alkaline environment, and then reacts with peptone in the medium The guanidine group contained in arginine acts to generate a red compound, which means that the V-P test is positive. If the content of guanidinium in the medium is low, a small amount of guanidinium-containing compounds such as creatine or creatinine can be added. This reaction can be accelerated by adding α-naphthol during the test.

[0057] 2) Medium

[0058] Peptone 5.0g, Glucose 5.0g, K 2 HPO 4 (or NaCl) 5.0 grams, 1000 milliliters of distilled water, pH 7.0-7.2, divided into test tubes, 5 milliliters per test tube, sterilized...

Embodiment 5

[0095] Drawing of Growth Curve of Bacillus subtilis K13 Strain

[0096] 1) method

[0097]Determination by colorimetry: put liquid LB medium into a 2000ml Erlenmeyer flask, sterilize and cool, insert activated Bacillus subtilis (Bacillus subtilis) K13 strain, place on a shaker at 28°C and 180 rpm nourish. Samples were taken every two hours, and the absorbance (OD600) of the bacterial solution at 600 nm was measured.

[0098] 2) Results:

[0099] figure 2 It is the growth curve graph of Bacillus subtilis (Bacillus subtilis) K13 strain, wherein OD600 is the ordinate, and the culture time is the abscissa. Because the absorbance is determined by the density of the bacterial solution, the growth trend of the bacteria can be inferred from the change trend of the absorbance.

[0100] Depend on figure 2 It can be seen that: 0~7h, Bacillus subtilis (Bacillus subtilis) K13 bacterial strain grows slowly, and is in stagnant growth phase; 7~16h, Bacillus subtilis (Bacillus subtilis...

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Abstract

The present invention discloses Bacillus subtilis K13 for inhibiting Colletotrichum gloeosporioides. The Bacillus subtilis K13 is preserved on May 27, 2015 at the China General Microbiological Culture Collection Center and has accession number CGMCC No. 10917, and belongs to the technical field of plant protection. The invention also discloses the culture method and application of the Bacillus subtilis K13; and the Bacillus subtilis K13 strain is obtained by a plate confrontation experiment screening, can effectively inhibit the growth of the Colletotrichum gloeosporioides; and the strain is harmless to human and livestock, green, secure and not easy to produce resistance, and has good environmental compatibility. The Bacillus subtilis K13 can be used in commercial development, and has broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of plant protection, and in particular relates to Bacillus subtilis K13 and its cultivation method and application. Background technique [0002] Of course, rubber is an important strategic material and a scarce resource. It is listed as the four major industrial raw materials together with iron ore, petroleum and coal. It plays a huge role in the transportation and defense industry, and it is closely related to the national economic situation, economic cycle, financial monetary policy and national trade policy; therefore, the rubber plantation industry has a pivotal position. The production process of natural rubber is affected by factors such as seasons and climate, just like agricultural products. In the agricultural development of hot areas in my country, in areas where natural rubber is relatively concentrated, local farmers and farm workers use natural rubber planting as a pillar of income increase. I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/125
Inventor 郑肖兰郑金龙贺春萍易克贤习金根吴伟怀梁艳琼李锐
Owner ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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