Saline-alkaline resistant bacillus megaterium FJW1 and application thereof to preparation of phytopathogen antibacterial agent

A technology of Bacillus megaterium and plant pathogenic bacteria, applied in the field of microorganisms, can solve problems such as limiting the scope of application

Active Publication Date: 2020-10-20
HUAZHONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently reported Bacillus megaterium that can be used as a biocontrol bacterium has only one or two types of inhibitory bacteria, which seriously limits its application range.

Method used

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  • Saline-alkaline resistant bacillus megaterium FJW1 and application thereof to preparation of phytopathogen antibacterial agent
  • Saline-alkaline resistant bacillus megaterium FJW1 and application thereof to preparation of phytopathogen antibacterial agent
  • Saline-alkaline resistant bacillus megaterium FJW1 and application thereof to preparation of phytopathogen antibacterial agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Isolation and identification of Bacillus megaterium FJW1:

[0048] Put 2.5g of soil samples from Pucheng County, Weinan City, Shaanxi into 50ml of sterile water, shake at 250rpm at 30°C for 1h, then take 500μl of soil suspension and carry out gradient dilution in a PA bottle filled with 4.5ml of sterile water. take 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 The soil suspensions of these dilutions were added to the culture medium for coating, and after 24h, 48h, 72h, and 96h of culture, the growth of plate colonies was observed, and all grown colonies were picked out for antibacterial activity experiments; Bacteria with high bacteriostatic rate and broad bacteriostatic spectrum were selected, and then further screened for saline-alkali tolerance; finally, Bacillus megaterium FJW1 was screened out.

[0049] (1) Morphological characteristics

[0050] The isolated saline-alkali-tolerant giant bacillus (Bacillus magaterium) FJW1, the colony morphology on LB solid medium is as f...

Embodiment 2

[0059] Tolerance test of Bacillus megaterium FJW1:

[0060] (1) pH range test:

[0061] The seed solution of Bacillus megaterium was inoculated into LB medium with different initial pH according to the inoculum amount of 1%, and cultured at 37°C and 200r / min. After 14 hours of culture, samples were taken to measure OD 600 . The initial pH gradient of the medium was set at 3, 4, 5, 6, 7, 8, 9, 10.

[0062] LB medium: peptone 10g, yeast extract powder 5g, NaCl 10g, distilled water 1000mL, pH 7.0, sterilized by high pressure steam at 121°C for 20min.

[0063] pH 3 4 5 6 7 8 9 10 OD 600

0.043 0.052 0.139 4.361 5.764 4.415 0.19 0.016

[0064] (2) Salt tolerance test:

[0065] The seed solution of Bacillus megaterium was inoculated into LB medium with different salt concentrations according to the amount of 1%, cultured at 37°C and 200r / min, and after 14 hours of culture, samples were taken to measure OD 600 . The salt concentration gradi...

Embodiment 3

[0072] Inhibitory effect of Bacillus megaterium FJW1 on plant pathogens:

[0073] The inhibitory ability of Bacillus megaterium to pathogenic bacteria was determined by zone of inhibition method. The activated Bacillus megaterium seed solution was appropriately diluted and spread on a PDA plate, and the sterile water was used as a control group. Punch holes on the PDA plate covered with pathogenic fungi, pick the bacterial block and inoculate it upside down on the PDA plate coated with Bacillus megaterium, make the side with the fungus touch the plate, and culture the plate at 37°C for 5 days. Then measure the fungal colony diameter, calculate the bacteriostatic rate, and then analyze the bacteriostatic effect.

[0074] The plant pathogens selected in this experiment are Fusarium oxysporum F.sp.Cubense, Fusarium oxysporum, Fusarium solani, Fusarium solani, Fusarium chlamydia Fusarium chlamydosporum, Penicillium glaucum, Rhizomucor variabilis, Banana wilt, Rhizopus nigricans,...

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Abstract

The invention belongs to the technical field of microorganisms and particularly relates to a bacillus megaterium FJW1 and an application thereof to preparation of a phytopathogen antibacterial agent.The invention relates to the bacillus megaterium FJW1, and the accession number is CCTCC M 2019660. The strain has alkali resistance, high environmental adaptability, phytopathogen resistance and other functions, can grow under the conditions of pH 6-8 and 13% salt concentration and has a relatively good inhibition effect on a plurality of fusarium pathogens, the antibacterial rate is up to 95%, the inhibition rate to banana fusarium wilt is 97% or more, the inhibition rate to penicillium glaucum and rhizomucor variabilis is 93% or more, the inhibition rate to rhizopus nigricans is 91% or more, the inhibition rate to cladosporium cladosporioides is 82% or more, the inhibition rate to botrytis cinerea and myrothecium roridum is 51% or more, the inhibition rate to alternaria tenuissima is 27% or more, and the bacillus megaterium FJW1 has potential application value of being made into a biocontrol bacterium.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a strain of bacillus megaterium FJW1 and its application in the preparation of antibacterial agents. Background technique [0002] The development and utilization of microbial fertilizers is an important way for the sustainable development of my country's agriculture. At this stage, the environmental pollution caused by the excessive use of pesticides, the resistance of diseases and insect pests, and the food security problems caused by pesticide residues are becoming more and more serious. Microbial pesticides have attracted much attention due to their advantages of green prevention and control, environmental friendliness, promotion of harmonious ecology, and protection of food safety. [0003] Bacillus preparations have attracted more and more attention due to their strong stress resistance and long-term storage. Compared with the vegetative body of bacteri...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/11
CPCA01N63/00C12N1/205C12R2001/11
Inventor 吴鹏宇陈振民任浩然王巧梅黄丽龚泽奇梁运祥
Owner HUAZHONG AGRICULTURAL UNIVERSITY
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