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Pseudo-scent-like aroma bacteria and its preparation method and application

A fragrance-like bacteria and fragrance-like technology, applied in the field of biological strains

Inactive Publication Date: 2017-06-20
TOBACCO RES INST CHIN AGRI SCI ACAD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the biological control of tobacco brown spot disease mostly focuses on the screening and utilization of microorganisms that produce non-volatile antimicrobial substances, while there is less research on microorganisms that produce volatile anti-tobacco brown spot metabolites

Method used

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  • Pseudo-scent-like aroma bacteria and its preparation method and application
  • Pseudo-scent-like aroma bacteria and its preparation method and application
  • Pseudo-scent-like aroma bacteria and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Medium: PDA medium: 200g potato, 20g glucose, 15g agar, 1000mL water.

[0029] LB solid medium: tryptone 10g, yeast extract 5g, sodium chloride 10g, agar 15g, water 1000mL.

[0030] The pathogenic bacteria tested: Alternaria alternate was provided by the Plant Protection Department of Tobacco Research Institute, Chinese Academy of Agricultural Sciences. Before the test, the mycelium pieces at the edge of the pathogenic bacteria stored in the refrigerator were picked and placed on fresh PDA medium for 7 days at 25°C for activation.

[0031] Bacterial strains to be tested: 67 bacterial strains were isolated from tobacco rhizosphere soil collected in Hunan Province by soil dilution method, numbered T1-T67, and stored in glycerol at -20°C. Before use, pipette the freshly thawed bacterial culture solution and spread it on LB medium, culture it at 28°C for 24 hours to activate, and streak to obtain a single colony. Before the buckle test, pick a single colony and incubate i...

Embodiment 2

[0032] Example 2 Preliminary Screening of Antagonistic Bacteria

[0033] The inhibitory effect of 67 strains of bacteria on Alternaria tabacum was tested by plate buckle method. Use a hole puncher with a diameter of 5 mm to punch holes in the medium covered with Alternaria tabacum to obtain mycelium blocks, inoculate them on a new PDA plate and culture them at 25°C for 12 hours to make them fix on the medium. Take 100 μL of the bacterial culture solution cultured by shaking for 24 hours and spread it evenly on the LB plate, place the PDA plate containing Alternaria Alternaria on the LB plate, and seal it with Parafilm. The LB plate without bacterial solution was used as a control. Seal the buckle plates with different treatments with fresh-keeping bags, and culture them at 25°C for 5-7 days. When the hyphae of Alternaria Alternaria in the control group cover 2 / 3 of the plates, use the cross method to measure the colony diameter of Alternaria Alternaria and calculate the volat...

Embodiment 3

[0036] The 16S rDNA identification of embodiment 3 bacteria

[0037] The screened bacteria with high inhibition rate were molecularly identified by 16S rDNA sequence determination and phylogenetic tree construction. Bacterial Genome Extraction Kit (Shanghai Sangong) was used to extract bacterial genomic DNA, and using this as a template, the 16S rDNA of the strain was analyzed with primers 27F: (5'-AGAGTTTGATCCTGGTCAGAACGCT-3') and 1492R: (5'-TACGGCTACCTTGTTACGACTTCACCCC-3'). Amplify. The obtained PCR product was purified and sent to Sangon Bioengineering Shanghai Co., Ltd. for sequence determination, see the sequence table below. The sequencing results were compared and analyzed by BLAST homology in the NCBI database, multiple sequence comparisons were performed with Clustal1.83, and a phylogenetic tree was constructed using the Neighbor-Joining method in Mega4.0.

[0038] Bacterial genomic DNA was extracted, amplified and then sent for sequencing to obtain its 16S rDNA seque...

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Abstract

The invention discloses a pseudo-aroma-like aroma bacteria T45 strain and its preparation and application. The pseudo-aroma-like aroma bacteria T45 strain produced by the invention produces volatile antibacterial substances, which have the effect of inhibiting the growth of tobacco red spot bacteria, mainly through Inhibition of conidia formation, spore germination, hyphal growth and destruction of mycelial morphology.

Description

technical field [0001] The invention relates to the technical field of biological strains, in particular to a pseudo-scent-like aroma bacterium, its preparation method and application. Background technique [0002] Alternaria alternate (Fries) Keissler is an important class of plant pathogenic bacteria with a wide range of hosts and serious damage. Tobacco red spot disease caused by it is an important leaf disease in the late mature stage of tobacco leaves. It is found in various smoking areas in my country. It directly affects the yield and quality of tobacco leaves. Regarding the prevention and control of tobacco red spot disease, chemical control is the main method in China at present, but the use of chemical pesticides to prevent and control red spot disease not only leads to pesticide residues, environmental pollution, but also causes drug resistance of bacteria and serious damage to tobacco plants. Because of its environmental friendliness, biological control is consid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01P3/00C12R1/01
CPCA01N63/10C12N1/20C12N1/205C12R2001/01
Inventor 孔凡玉黄树立游偲李锡宏张成省李青诚陈德鑫高加明冯超王静
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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