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Bacillus methylotrophicus F7 and application thereof

A methylotrophic and Bacillus technology, applied in the direction of application, bacteria, and fungicides, can solve problems such as difficult to achieve ideal effects, and achieve high-efficiency biological control capabilities and significant biological control effects

Pending Publication Date: 2017-05-31
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Many strains often show antagonistic activity in plate experiments, but it is difficult to have ideal effects in pot and field stages

Method used

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  • Bacillus methylotrophicus F7 and application thereof
  • Bacillus methylotrophicus F7 and application thereof
  • Bacillus methylotrophicus F7 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Screening and Purification of Bacterial Strains

[0047] (1) Primary screening

[0048] Beef extract-peptone medium: 10g bacteriological peptone, 3g beef extract, 5g sodium chloride, 18g agar, 1000mL distilled water, adjust the pH to 7.0-7.2.

[0049] Pseudomonas solanacearum (Ralstonia Solanacearum) was used as the pathogen indicator bacteria to screen antagonistic strains. Streak Pseudomonas solanacearum on a TTC plate, culture it in a constant temperature incubator at 30°C for 48 hours, and then observe it, and pick the strains with strong pathogenicity and cultivate them for later use. Take the culture solution of Pseudomonas solanacearum cultured at 30°C for 24 hours, use a pipette to draw 200 μL of the bacteria solution into the sterile plate, pour it into the plate containing the bacteria solution, and cool it to about 45°C About 20mL of NA medium per plate was placed in a horizontal position and the plate was quickly rotated to mix the medium and the...

Embodiment 2

[0055] Example 2 Characterization

[0056] (1) The morphological characteristics of the bacteria are Gram-positive bacteria with spores. The bacteria are rod-shaped, motile and aerobic. Spore staining photos of bacteria see figure 1 .

[0057] (2) The morphological characteristics of the colony. The colonies formed after culturing on the beef extract peptone medium for 24 hours are round or irregular. After 48 hours, the colonies are all round, milky white, about 1-2 mm in diameter, with irregular edges and moist . F7 bacteria have swimming and wriggling properties, and spread to the entire plate after being cultured overnight on LB medium containing 0.3% agar, indicating that F7 bacteria have swimming properties (swim), and overnight culture on LB medium containing 0.7% agar has a certain Swimming (swam), see figure 2 .

[0058] (3) Growth characteristics, cultured for 18 hours under conditions of 5g of yeast extract powder, 10g of peptone, 10g of sodium chloride, and ...

Embodiment 3

[0065] Example 3 Experiment of siderophore production by F7 bacteria

[0066] Transfer the strain to the LB plate and culture it for 24-48 hours, use the point inoculation method, use a sterilized toothpick to inoculate the strain on the CAS solid detection plate, spot 3 replicates, culture at 28°C for 3 days, and test the cultured CAS On the plate, due to the iron ions chelated by EDTA in the siderophilic competition medium, the medium changes from blue to yellow, thereby judging whether siderophores are produced. Test results such as Figure 4 As shown in , an obvious yellow halo appeared around the strain, indicating that the strain produced siderophore.

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Abstract

The invention discloses bacillus methylotrophicus F7 and an application thereof. The invention aims at providing a strain which has a plurality of beneficial properties, namely producing siderophore, cellulase and protease, producing auxin, producing ammonia and the like; the strain not only can be used for effectively inhibiting various pathogenic bacteria such as ralstonia solanacearum, fusarium oxysporum f.sp.cubense, colletotrichum gloeosporioides, fusarium oxysporum f.sp cucumerinum but also can promote plant growth. In accordance with the technical scheme, the bacillus methylotrophicus F7 is preserved in China General Microbiological Culture Collection Center (CGMCC) on February 1, 2016, with preservation number of CGMCC No.12124; in addition, five surfactin homologs, six fengycin homologs and three iturin homologs, which have an antibacterial effect, are extracted from fermentation liquid of the bacillus methylotrophicus F7; and the bacillus methylotrophicus F7 belongs to the field of agricultural biotechnology.

Description

technical field [0001] The invention relates to the field of biological control of plant soil-borne diseases, and relates to a microorganism and its application, in particular to a methylotrophic bacillus F7 (Bacillus methylotrophicus) and its application. Background technique [0002] Tomato bacterial wilt is a soil-borne bacterial wilt disease caused by Ralstonia solanacearum infecting tomato tissue, which seriously threatens the yield and quality of tomato worldwide. For a long time, chemical pesticides have been used as the main means to control tomato bacterial wilt, but there are many disadvantages such as few varieties of pesticides and unsatisfactory control effects. At the same time, it has also caused a series of problems such as ecological environment damage, food pollution, and human and animal poisoning. However, due to the difficulty in developing new high-efficiency fungicides and the long breeding cycle of resistant varieties, in recent years, biocontrol tech...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P21/00A01N63/00A01P1/00A01P3/00A01P21/00C12R1/07
CPCA01N63/00C12P21/00C12N1/205C12R2001/07
Inventor 王玉琪裴秦汉蔡燕飞李永涛熊汉琴曹禺周晗
Owner SOUTH CHINA AGRI UNIV
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