Pseudomonas aeruginosa and application thereof

Abstract

The invention provides pseudomonas aeruginosa. A classification name of the pseudomonas aeruginosa is pseudomonas aeruginosa T1-3, the pseudomonas aeruginosa is preserved in the Common Microorganism Center of the China Committee for Culture Collection of Microorganisms, the address is the Institution of Microbiology of the Chinese Academy of Sciences in the No.3 yard of the No.1 Beichen West Road in the Chaoyang district of Beijing, the postal code is 100101, a preservation number is CGMCC No.10433, and a preservation date is January 23, 2015. The pseudomonas aeruginosa has the advantages that excellent prevention and treatment effects can be realized by the pseudomonas aeruginosa for diversified diseases of plants such as corn, cow peas, chili, cabbage and leek, and the pseudomonas aeruginosa has wide antibacterial spectra; the pseudomonas aeruginosa is high in reproduction speed and stress resistance and favorable for production application, and can be artificially cultivated, and cultivation conditions are simple; the pseudomonas aeruginosa is free of pathogenicity to people and livestock, is safe and effective and accordingly has an excellent development prospect.

Description

technical field

[0001] The invention relates to the field of microbes, in particular to a kind of Pseudomonas aeruginosa, and also relates to the application of the Pseudomonas aeruginosa in preventing and treating plant diseases. Background technique

[0002] Fungi are important pathogens that cause plant diseases. After infecting the host, crop roots, stems, leaves, flowers, fruits and seeds will produce discoloration, necrosis, rot, wilting or deformity symptoms. At present, diseases of crops such as corn, cowpea, and green onion are mainly controlled by chemical fungicides. On the one hand, these agents have potential harm to the environment and human health, and on the other hand, they will increase the resistance of pathogenic fungi. Therefore, relying solely on Using chemical fungicides to control fungal diseases is not a long-term solution for environmental protection.

[0003] Antagonistic bacteria can inhibit the growth of pathogenic bacteria by secreting and prod...

Examples

Embodiment 1

[0028] The isolation and screening of embodiment 1 Pseudomonas aeruginosa bacterial strain

[0029] 1. Medium:

[0030] Potato dextrose agar medium (PDA medium): 200g of potatoes, 20g of glucose, 15-20g of agar, natural pH, and dilute to 1L.

[0031] Beef extract peptone medium (LB medium): tryptone 10g, yeast extract powder 5g, sodium chloride 10g, neutral pH, dilute to 1L.

[0032] 2. Isolation and screening of Pseudomonas aeruginosa strains

[0033] A) take by weighing the rhizosphere soil 10g of the healthy maize plant that does not have this disease to take place near the more serious region of maize leaf spot disease incidence, add 90ml sterile water, get 10 -1 Dilution, in a shaker at 30°C at 130r / min, cultivated for 2-3h, continued to add sterile water to dilute to obtain 10 -5 , 10 -6 , 10 -7 , 10 -8 Take 200ml of the above dilution solution on LB plates, culture at 30°C for 48 hours, pick a single colony and inoculate it on the slope of LB solid medium for stor...

Embodiment 2

[0048] Embodiment 2T1-3 bacterial strain affects the growth of cowpea rust pathogen under liquid culture

[0049] Liquid medium preparation method: corn flour 10g, soybean flour 5g, glucose 5g, peptone 2g, NaCl 1g, K 2 HPO 4 1g, MgS0 4 ·7H 2 O0.2g, dilute to 1L, natural pH.

[0050] Martin's plate medium: peptone 5g, glucose 10g, K 2 HPO 4 1g, MgS0 4 ·7H 2 O0.5g, dilute to 1L, natural pH, add 3.3mL of 1% Bengal red aqueous solution. When in use, add 0.3mL of 1% streptomycin solution per 100mL of culture medium to make plate culture medium.

[0051] experiment method:

[0052] (1) Cultivation in liquid medium: inoculate Pseudomonas aeruginosa T1-3 in liquid medium, 30°C, shaker speed 150r / min, 48h; the control group inoculates 3 cowpea rust bacteria dishes, The treatment group was simultaneously inoculated with 3 cowpea rust germ plates and 100uL of liquid culture solution comprising Pseudomonas aeruginosa (Pseudomonasaeruginosa) T1-3; the results are shown in image...

Embodiment 3T1-3

[0056] The preparation of embodiment 3T1-3 bacterial strain inoculum

[0057] Pseudomonas aeruginosa (Pseudomonasaeruginosa) T1-3 was inoculated into LB liquid medium, and cultured with shaking at 30°C and 130r / min for 24h to form seed liquid.

[0058] Then inoculate the seed liquid into the culture material in the fermenter at a volume ratio of 1:400 for fermentation production. The basic working parameters of the fermenter are: temperature: 30-35°C, stirring speed: 130r / min, pH: 6.8- 7.2, dissolved oxygen: 30%, tank pressure: 0.04MPa. After 24 hours, stop the fermentation, then centrifuge at 6000r / min for 10min, take the sediment and dilute it with sterile water to make a bacterial agent. The concentration of viable bacteria in the bacteria agent was detected to be 1×10 9 -1×10 10 CFU / mL.

[0059] The prepared bacterial agent can be used for biological control of common corn diseases, cowpea rust and other diseases.

[0060]

[0061]

[0062]

technical field

[0001] The invention relates to the field of microbes, in particular to a kind of Pseudomonas aeruginosa, and also relates to the application of the Pseudomonas aeruginosa in preventing and treating plant diseases. Background technique

[0002] Fungi are important pathogens that cause plant diseases. After infecting the host, crop roots, stems, leaves, flowers, fruits and seeds will produce discoloration, necrosis, rot, wilting or deformity symptoms. At present, diseases of crops such as corn, cowpea, and green onion are mainly controlled by chemical fungicides. On the one hand, these agents have potential harm to the environment and human health, and on the other hand, they will increase the resistance of pathogenic fungi. Therefore, relying solely on Using chemical fungicides to control fungal diseases is not a long-term solution for environmental protection.

[0003] Antagonistic bacteria can inhibit the growth of pathogenic bacteria by secreting and prod...

Examples

Embodiment 1

[0028] The isolation and screening of embodiment 1 Pseudomonas aeruginosa bacterial strain

[0029] 1. Medium:

[0030] Potato dextrose agar medium (PDA medium): 200g of potatoes, 20g of glucose, 15-20g of agar, natural pH, and dilute to 1L.

[0031] Beef extract peptone medium (LB medium): tryptone 10g, yeast extract powder 5g, sodium chloride 10g, neutral pH, dilute to 1L.

[0032] 2. Isolation and screening of Pseudomonas aeruginosa strains

[0033] A) take by weighing the rhizosphere soil 10g of the healthy maize plant that does not have this disease to take place near the more serious region of maize leaf spot disease incidence, add 90ml sterile water, get 10 -1 Dilution, in a shaker at 30°C at 130r / min, cultivated for 2-3h, continued to add sterile water to dilute to obtain 10 -5 , 10 -6 , 10 -7 , 10 -8 Take 200ml of the above dilution solution on LB plates, culture at 30°C for 48 hours, pick a single colony and inoculate it on the slope of LB solid medium for stor...

Embodiment 2

[0048] Embodiment 2T1-3 bacterial strain affects the growth of cowpea rust pathogen under liquid culture

[0049] Liquid medium preparation method: corn flour 10g, soybean flour 5g, glucose 5g, peptone 2g, NaCl 1g, K 2 HPO 4 1g, MgS0 4 ·7H 2 O0.2g, dilute to 1L, natural pH.

[0050] Martin's plate medium: peptone 5g, glucose 10g, K 2 HPO 4 1g, MgS0 4 ·7H 2 O0.5g, dilute to 1L, natural pH, add 3.3mL of 1% Bengal red aqueous solution. When in use, add 0.3mL of 1% streptomycin solution per 100mL of culture medium to make plate culture medium.

[0051] experiment method:

[0052] (1) Cultivation in liquid medium: inoculate Pseudomonas aeruginosa T1-3 in liquid medium, 30°C, shaker speed 150r / min, 48h; the control group inoculates 3 cowpea rust bacteria dishes, The treatment group was simultaneously inoculated with 3 cowpea rust germ plates and 100uL of liquid culture solution comprising Pseudomonas aeruginosa (Pseudomonasaeruginosa) T1-3; the results are shown in image...

Embodiment 3T1-3

[0056] The preparation of embodiment 3T1-3 bacterial strain inoculum

[0057] Pseudomonas aeruginosa (Pseudomonasaeruginosa) T1-3 was inoculated into LB liquid medium, and cultured with shaking at 30°C and 130r / min for 24h to form seed liquid.

[0058] Then inoculate the seed liquid into the culture material in the fermenter at a volume ratio of 1:400 for fermentation production. The basic working parameters of the fermenter are: temperature: 30-35°C, stirring speed: 130r / min, pH: 6.8- 7.2, dissolved oxygen: 30%, tank pressure: 0.04MPa. After 24 hours, stop the fermentation, then centrifuge at 6000r / min for 10min, take the sediment and dilute it with sterile water to make a bacterial agent. The concentration of viable bacteria in the bacteria agent was detected to be 1×10 9 -1×10 10 CFU / mL.

[0059] The prepared bacterial agent can be used for biological control of common corn diseases, cowpea rust and other diseases.

[0060]

[0061]

[0062]

Images