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Fengycin, composition including fengycin, fusion gene encoding fengycin, recombinant plasmid, recombinant bacteria and applications

A technology of fusing genes and recombinant plasmids, which is applied in the fields of Feng Yuan, recombinant bacteria and applications, can solve the problems of poor control effect, etc., and achieve the effect of obvious inhibitory effect and high-efficiency biocontrol effect.

Active Publication Date: 2021-03-19
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the use of highly toxic pesticides for many years, pathogenic bacteria have developed resistance to pesticides, and the control effect has deteriorated.

Method used

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  • Fengycin, composition including fengycin, fusion gene encoding fengycin, recombinant plasmid, recombinant bacteria and applications
  • Fengycin, composition including fengycin, fusion gene encoding fengycin, recombinant plasmid, recombinant bacteria and applications
  • Fengycin, composition including fengycin, fusion gene encoding fengycin, recombinant plasmid, recombinant bacteria and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Extraction of lipopeptides produced by Bacillus velei HN-Q-8

[0060] Use the cross-hatching method to culture a single colony at 37°C for 24h, inoculate the newly activated HN-Q-8 strain in 100mL LB liquid medium for 24h at 37°C and 200r / min to obtain a seed liquid; inoculate the seed liquid to In 100mL LB liquid medium, the inoculum amount was 10%, and continued to cultivate at 37°C and 200r / min for 72h: Centrifuge the fermentation broth at 4°C and 8000rpm for 20min, collect the supernatant, and slowly add 6mol / Adjust the pH to 2.0 with L of HCl, and let it settle overnight at 4°C; then, centrifuge at 8,000 rpm for 20 minutes at 4°C to collect the precipitate, dry it in a fume hood, and then add 30 mL of methanol to fully dissolve the precipitate. Filter the methanol extract with a bacterial filter to obtain lipopeptide compounds.

Embodiment 2

[0061] The antibacterial activity of the lipopeptide compound that embodiment 2 Bacillus Velez HN-Q-8 produces

[0062] The potato black nevus strain ZB4 was used as the model bacteria, and the concentration of the lipopeptide compound was 10mg / mL, 5mg / mL, 1mg / mL, 0.5mg / mL, 0.1mg / mL. Insert the model pathogenic bacteria cake (5 mm) into the center of the plate, punch holes on both sides 2.5 cm away from the bacteria cake, add 50 μL of methanol to one side, and add 50 μL of lipopeptide compound to the other side. Each concentration is a treatment, and each treatment is repeated 3 times, and an untreated blank plate is used as a control. When the blank control group is about to cover the petri dish, observe and take pictures.

[0063] The result is as figure 1 as shown, figure 1 In A, the concentration is 10mg / mL lipopeptide compound antibacterial activity; B is 5mg / mL lipopeptide compound antibacterial activity; C is 1mg / mL lipopeptide compound antibacterial activity; D is 0...

Embodiment 3

[0064] The lipopeptide compound bacteriostatic spectrum measurement that embodiment 3 Velei bacillus HN-Q-8 produces

[0065] 1. Determination of the inhibition of lipopeptide compounds on pathogens: the concentration used in the determination of the bacteriostatic spectrum is 10 mg / mL. Insert fungal pathogens (5 mm) into the center of the plate, punch holes on both sides 2.5 cm away from the bacteria cake, add 50 μL of methanol to one side, and add 50 μL of lipopeptide compound to the other side. Each bacterium is a treatment, and each treatment is repeated three times, and an untreated blank plate is used as a control. When the blank control group covered the Petri dish, observe and take pictures.

[0066] 2. Determination of the inhibition of lipopeptide compounds on bacteria and actinomycetes: the dilution factor is 1 × 10 -5 Potato blackleg and the dilution factor was 1×10 -2 100 μL of the bacterial solution of potato scab bacteria was evenly spread on the plate, and t...

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Abstract

The invention provides fengycin, a fusion gene encoding lipopeptide compounds, recombinant plasmid, recombinant bacteria and applications, and belongs to the technical field of plant disease control.The fengycin is produced by Bacillus velezensis HN-Q-8; and the accession number of the Bacillus velezensis HN-Q-8 is CGMCC No. 19554. The Bacillus velezensis HN-Q-8 has efficient biocontrol effects.In the inhibitory substances produced by the Bacillus velezensis HN-Q-8, the fengycin plays a principal role of bacteriostasis. The fengycin has obvious inhibition effects on potato black scurf.

Description

technical field [0001] The invention belongs to the technical field of plant disease prevention and control, and particularly relates to fengyuansu, a composition including fengyuansu, a fusion gene encoding fengyuansu, a recombinant plasmid, a recombinant bacterium and applications. Background technique [0002] Potato black scurf is caused by Rhizoctonia solani. In recent years, with the further increase of potato planting area in my country, the occurrence and damage of this disease have become increasingly serious due to the inability to realize crop rotation in many main potato producing areas, and the appearance and commodity value of potato pieces have been severely damaged. At present, it mainly relies on chemical agents for prevention and control. However, due to the use of highly toxic pesticides for many years, pathogenic bacteria have developed resistance to pesticides, and the control effect has deteriorated. Therefore, biological control has become the focus ...

Claims

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Application Information

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IPC IPC(8): C07K14/32C12N15/32C12N15/62C12N15/70C12N1/21A01N37/46A01N63/20A01N63/60A01P3/00C12R1/19
CPCC07K14/32C12N15/70A01N37/46A01N63/20A01N63/60C07K2319/00A01N63/22
Inventor 朱杰华杨志辉赵冬梅杨毅清赵雅
Owner HEBEI AGRICULTURAL UNIV.
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