Bacillus subtilis Baisha2C for restraining plant pathogenic fungi

A technology for Bacillus subtilis and plant fungal diseases, which is applied in the directions of plant growth regulators, botanical equipment and methods, and microorganism-based methods to achieve the effects of stable antibacterial ability, simple cultivation and strong adaptability

Active Publication Date: 2015-09-30
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

However, there are relatively few reports on the induction of plant resistance by Bacillus, and studies have

Method used

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  • Bacillus subtilis Baisha2C for restraining plant pathogenic fungi
  • Bacillus subtilis Baisha2C for restraining plant pathogenic fungi
  • Bacillus subtilis Baisha2C for restraining plant pathogenic fungi

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Isolation, cultivation, screening and purification of biocontrol bacteria Baisha2C.

[0027] 1.1 Sampling

[0028] The samples were the susceptible leaves of pepper in Baisha County, Hainan Province. The collected leaves were put into plastic bags and recorded.

[0029] 1.2 Surface disinfection of materials and separation of antagonistic bacteria

[0030] 1.2.1 Rinse with clean water and cut off diseased leaves

[0031] Wash off the dust and grit on the leaves with clean water and let them dry. Use scissors to cut small leaves with a size of 5 × 5mm at the junction of disease and health for future use.

[0032] 1.2.2 Sample disinfection

[0033] Put the clipped small diseased leaves into a beaker and soak them in mercuric chloride for 30 seconds. During the soaking process, pay attention to constantly stirring the small diseased leaves, then rinse them with sterile water three times, dry them, and set them aside.

[0034] 1.2.3 Bacterial isolation and culture

[0...

Embodiment 2

[0042] Identification of Biocontrol Bacteria Baisha2C

[0043] 1. Microbiological characteristics:

[0044] Observing the morphological characteristics of the biocontrol strains cultivated by streaking in the incubator at 37°C, it was found that the biocontrol strain Baisha2C was rod-shaped, tended to form chains, and had round or square ends; spores were oval, oval, columnar, and round , without a capsule, belonging to the genus Bacillus. After Gram staining, the bacteria showed blue-purple, which was Gram-positive bacteria.

[0045] 2. Molecular biological identification:

[0046] (1) DNA extraction: Genomic DNA was extracted using a general kit in the art;

[0047] (2) PCR reaction: bacterial general primers 8F and 1492R were synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd., and the structure is as follows: 8F 5'-AGAGTTTGATCCTGGCTCAG-3', 1492R5'-GGTTACCTTGTTACGACTT-3'.

[0048] The 16S rDNA gene amplification results of the biocontrol strain Baisha2C are as fol...

Embodiment 3

[0130] Study on Sensitivity of Biocontrol Bacteria Baisha2C to Antibiotics

[0131] 1) culture medium

[0132] LB solid medium.

[0133] 2) Reagent

[0134] Ampicillin, kanamycin, chloramphenicol, streptomycin, tetracycline, erythromycin, rifampicin, cephalosporin.

[0135] 3) method

[0136] The sensitivity of Bacillus subtilis K13 to non-antibiotics was determined by the filter paper method, and a filter paper disc with a diameter of 7 mm was prepared with a puncher and sterilized at high temperature. Add 1 ml of the seed solution of the tested strain to the LB solid medium cooled to 50-55 degrees Celsius after melting, shake well and pour the plate, put the sterilized filter paper on the plate, and then add the antibiotic solution on the filter paper The amount added is equivalent to ampicillin Amp 50 micrograms per milliliter, kanamycin Kana 50 micrograms per milliliter, chloramphenicol Chl 50 micrograms per milliliter, streptomycin Str 50 micrograms per milliliter, te...

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Abstract

The invention belongs to the field of biocontrol strain excavation and utilization, in particular to bacillus subtilis Baisha2C for restraining plant pathogenic fungi, and the preservation number of the bacillus subtilis Baisha2C is CGMCC No.10963. The invention further discloses molecular biological characteristics, physiological-biochemical characteristics and a cultivation method of the bacillus subtilis Baisha2C, and the bacillus subtilis Baisha2C has activity for restraining various kinds of the plant pathogenic fungi, such as fusarium colletotrichum, colletotrichum gloeosporioides and fusarium.

Description

technical field [0001] The invention relates to the field of mining and utilization of biocontrol strains, in particular to the identification and application of a strain of Bacillus subtilis Baisha2C that inhibits plant pathogenic fungi. Background technique [0002] Garratt, S.D. (1965), a British expert on plant fungal diseases, pointed out that "biological control is to reduce the survival and activity of pathogens through the action of other organisms (except humans) under any conditions or by means of any measures, thereby reducing pathogens. Occurrence of diseases”; “Biological control refers to the introduction of one or several kinds of organisms for control, by increasing the number of the organisms or by changing the environment to facilitate the activities of these organisms, usually a combination of the two measures”. [0003] Biological control of plant diseases is the use of one or more antagonists to reduce the density or pathogenic activity of pathogenic bac...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P3/00C12R1/125
CPCA01N63/00A01N63/10C12N1/205C12R2001/125
Inventor 郑肖兰梁艳琼贺春萍习金根易克贤李锐吴伟怀郑金龙秦周
Owner ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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