CN116391727A - Application of shenqinmycin and pseudomonas in prevention and treatment of soil-borne diseases - Google Patents
Application of shenqinmycin and pseudomonas in prevention and treatment of soil-borne diseases Download PDFInfo
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/27—Pseudomonas
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/48—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with two nitrogen atoms as the only ring hetero atoms
- A01N43/60—1,4-Diazines; Hydrogenated 1,4-diazines
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Environmental Sciences (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Virology (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to the technical field of pesticide compounding, in particular to application of a compound of shenzinomycin and pseudomonas in preventing and treating soil-borne diseases. The invention provides a compound biological pesticide for preventing and treating soil-borne diseases, and a preparation method and application thereof, wherein the biological pesticide is prepared by compounding a pseudomonas agent and promethazine according to a proportion, and the pseudomonas comprises the following components: one or more of Pseudomonas rape, pseudomonas fluorescens and Pseudomonas viridis. The shenqinmycin can promote the growth of crops, has good control effect on various soil-borne diseases, and the pseudomonas has the effects of inhibiting the growth of pathogenic organisms by using synthetic antibiotics, promoting the growth of crops and inducing plants to generate systemic resistance, thereby having good application prospect in the field of biological pesticides.
Description
Technical Field
The invention relates to the technical field of pesticide compounding, in particular to application of a compound of shenzinomycin and pseudomonas in preventing and treating soil-borne diseases.
Background
Soil-borne diseases refer to crop diseases caused by infection of plant roots, stems by plant pathogenic fungi, bacteria, viruses and nematodes living in soil under proper conditions. Soil-borne diseases are extremely common, and common soil-borne diseases include banded sclerotial blight of rice, corn and wheat, cotton damping off, verticillium wilt, soybean root rot and the like. Pathogenic bacteria in soil that cause soil-borne diseases include the genus Pythium (Pythium spp.); fusarium (Fusarium), rhizoctonia, verticillium (Verticillium) and the like cause seedling damping-off and wilt, wilt and Verticillium and the like. In recent years, with the improvement of intensive planting system, soil-borne diseases caused by factors such as poor management, continuous planting of single crops, returning of straws to fields and the like are increasingly serious, high yield, stable yield and agricultural product quality of crops are seriously affected, and the soil-borne diseases become an important bottleneck for limiting sustainable development of agriculture.
The 'pre-prevention and comprehensive control' is a plant protection policy established in China from the 70 th century of 20 th, and the biological control utilizes multiple action mechanisms of beneficial microorganisms on plant pathogenic bacteria to weaken, kill, antagonize and compete for the pathogenic bacteria. The biological control measures are utilized to promote the propagation of beneficial microorganisms in soil, decompose pesticide and chemical fertilizer remained in the soil, regulate the pH value of the soil and effectively prevent soil-borne diseases. The biggest advantages of biological control are that the biological control is environment-friendly, safe to human body, beneficial to plant growth, alleviating drug resistance generated by pathogenic bacteria, and lasting control effect. Biological control has become an important way of controlling crop diseases. The research shows that pseudomonas fluorescens, pseudomonas brassicae, pseudomonas viridis and the like of the pseudomonas have good inhibition effect on the growth of various soil-borne disease pathogenic bacteria. The microbial agent has high safety and is environment-friendly, and the pseudomonas prepared into the microbial agent has better application potential for preventing and treating soil-borne diseases.
The promethazine can promote the growth of crops, and the mechanism of the antibacterial activity of the promethazine has at least two aspects: (1) In the process that the shenqimycin is reduced in pathogenic bacteria cells, toxic superoxide ions and hydrogen peroxide can be generated, and glutathione and transferrin can be oxidized, so that hydroxyl free radicals with high toxicity to cells are generated; (2) Since the shenqimycin can be reduced by NADH, it becomes an intermediate of electron transfer, and the normal redox homeostasis (NADH/NAD + Ratio, etc.), to influence the generation of energy, thereby inhibiting the growth of microorganisms.
The pseudomonas can compete with pathogenic bacteria for nutrients and inhibit the growth of the pathogenic bacteria; influence the metabolism of host plants to change the physiological characteristics of the plants and inhibit the growth of pathogenic bacteria; enhancing the stress resistance of the host plant to low temperature, drought, pesticide, salt and alkali, environmental pollution and other stresses, and maintaining the normal growth of the host plant under the stress condition; improving the repair capacity of the host plant; promote the host plant to produce secondary metabolite and antagonize and induce resistance and other disease preventing mechanisms through competition. The effects of promoting growth and resisting diseases are achieved through the methods.
The shenqinmycin serving as a broad-spectrum bactericide has control effect on various soil-borne diseases, but the shenqinmycin has higher price, and the control effect on single diseases is lower than that of chemical agents. When the pseudomonas is singly applied, two problems exist, namely, the colonization of the pseudomonas at the plant rhizosphere is easily influenced by external environment factors, and a certain time is needed from the colonization of the rhizosphere to the exertion of the biocontrol effect. The biological pesticides with different action modes are reasonably compounded, and the proper use method and period are combined, so that the dosage of the biological pesticides is reduced and the control effect is improved. Therefore, the compound pesticide of the shenqimycin and the pseudomonas has important practical significance.
Disclosure of Invention
In view of the above, the application of the compound of the shenqimycin and the pseudomonas in preventing and controlling soil-borne diseases can play a role in inhibiting bacteria efficiently and rapidly, overcome the defect of slow effect caused by independently applying the pseudomonas, reduce the dosage of the shenqimycin, reduce the cost, delay the generation of drug resistance and improve the preventing and controlling effect.
In order to achieve the above object, the present invention provides the following technical solutions:
the present invention provides compositions, including pseudomonas and promethazine.
In some embodiments of the present invention, the volume ratio of said Pseudomonas to said Shenqimycin of the above composition is (1-19): 1.
In some embodiments of the invention, the pseudomonas of the above composition comprises one or more of pseudomonas brassicae, pseudomonas fluorescens, or pseudomonas aeruginosa.
In some embodiments of the invention, in the above composition:
the preservation number of the pseudomonas brassicae is CGMCC No.6774; and/or
The preservation number of the pseudomonas fluorescens is CGMCC No.0462; and/or
The preservation number of the pseudomonas aeruginosa is CGMCC No.6773.
In some embodiments of the invention, in the above composition:
the viable count of the pseudomonas brassicae is 100 hundred million CFU/mL to 190 hundred million CFU/mL; and/or
The viable count of the pseudomonas fluorescens is 75 hundred million CFU/mL to 140 hundred million CFU/mL; and/or
The living bacteria number of the pseudomonas aeruginosa is 75 hundred million CFU/mL to 140 hundred million CFU/mL.
In some embodiments of the invention, the culture medium for culturing the pseudomonas brassicae, the pseudomonas fluorescens, and/or the pseudomonas aeruginosa in the above composition comprises a KB culture medium.
In some embodiments of the present invention, in the above composition, the fermentation medium of pseudomonas rape comprises 3% sucrose, 1% yeast powder, 0.5% potassium chloride, and the balance water.
In some embodiments of the invention, in the above composition, the culture conditions of pseudomonas brassicae include: ph=7; the temperature is 28-30 ℃; the rotating speed is 180-200 r/min; 24-30 h.
In some embodiments of the present invention, the fermentation medium of Pseudomonas fluorescens comprises corn flour 2%, bean cake flour 4%, peptone 1.5%, K in the above composition 2 HPO 4 0.02％、KH 2 PO 4 0.03%, anhydrous magnesium sulfate 0.06%, glycerin 0.3%, and the balance being water.
In some embodiments of the invention, in the above composition, the culture conditions of the Pseudomonas fluorescens include: ph=7; the temperature is 35-37 ℃; the rotating speed is 180-200 r/min; 24-30 h.
In some embodiments of the present invention, the fermentation medium of Pseudomonas aeruginosa comprises yeast extract 0.5%, peptone 1%, sodium chloride 0.5%, na 2 HPO 4 0.02％、KH 2 PO 4 0.03%, anhydrous magnesium sulfate 0.1%, and the balance being water.
In some embodiments of the invention, in the above composition, the culture conditions of the pseudomonas aeruginosa include: ph=7; the temperature is 28-30 ℃; the rotating speed is 180-200 r/min; 24-30 h.
In some embodiments of the invention, in the above composition, the pseudomonas brassicae comprises a pseudomonas brassicae agent comprising a pseudomonas brassicae fluid, diatomaceous earth, and a protective agent;
the proportion of the pseudomonas liquid for rape, the diatomite and the protective agent is (10-7): 4-2): 2-1;
the protective agent comprises 1% sodium alginate;
the viable count of the pseudomonas brassicae microbial inoculum is 200 hundred million CFU/g.
In some embodiments of the invention, in the above composition, the pseudomonas fluorescens comprises a pseudomonas fluorescens agent comprising a pseudomonas fluorescens liquid, diatomaceous earth, and a protective agent;
the proportion of the pseudomonas fluorescens liquid, the diatomite and the protective agent is (10-7): 4-2): 2-1;
the protective agent comprises 1% sodium alginate;
the viable count of the Pseudomonas fluorescens bacterial agent is 150 hundred million CFU/g.
In some embodiments of the invention, in the above composition, the pseudomonas aeruginosa comprises a pseudomonas aeruginosa agent comprising pseudomonas aeruginosa liquid, diatomaceous earth and a protecting agent;
the ratio of the pseudomonas aeruginosa liquid, the diatomite and the protective agent is (10-7): 4-2): 2-1;
the protective agent comprises 1% sodium alginate;
the viable count of the pseudomonas aeruginosa agent is 150 hundred million CFU/g.
In some embodiments of the present invention, the recited promethazine is used at a concentration of 1% (v/v) in the above-described compositions.
The invention also provides application of the composition in preventing and treating soil-borne diseases;
the soil-borne disease comprises one or more of wheat scab, rice sheath blight, pepper blight, watermelon fusarium wilt, cucumber gray mold, apple ring rot, tomato fusarium wilt, corn sheath blight, cotton fusarium wilt, melon gummy stem blight, potato late blight or tomato gray mold.
The invention also provides application of the composition in preparing pesticides for preventing and treating soil-borne diseases;
the soil-borne disease comprises one or more of wheat scab, rice sheath blight, pepper blight, watermelon fusarium wilt, cucumber gray mold, apple ring rot, tomato fusarium wilt, corn sheath blight, cotton fusarium wilt, melon gummy stem blight, potato late blight or tomato gray mold.
The invention also provides a pesticide, which comprises the composition and acceptable auxiliary materials or auxiliary agents.
In some embodiments of the invention, the pesticide described above comprises the pesticide after 500-fold dilution.
The invention also provides a preparation method of the pesticide, which comprises the step of diluting the composition by 500 times to obtain the pesticide.
The invention also provides a using method of the pesticide, which comprises soil treatment, leaf surface spraying, flushing, root irrigation, root dipping and the like.
The composition and the application of the composition have the following effects:
1) Can prevent and treat various soil-borne diseases and promote the growth of crops;
2) By compounding the pseudomonas with different action mechanisms and periods and the shenqimycin, the application sites of pathogenic bacteria can be increased, the synergy is realized, the control effect is improved, the high-efficiency and rapid antibacterial effect of the shenqimycin can be exerted, the defect of slow effect of independently applying the pseudomonas agent is overcome, the dosage of the shenqimycin can be reduced, the cost is reduced, the generation of drug resistance is delayed, and the control effect is improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 shows an in-vessel bacteriostasis test of watermelon fusarium wilt; wherein A shows a control, B shows a pseudomonas brassicae fermentation broth, C shows a promethazine, and D shows a pseudomonas brassicae fermentation broth compounded promethazine;
FIG. 2 shows an in-vessel bacteriostasis test of rice sheath blight disease; wherein A shows a control, B shows a pseudomonas brassicae fermentation broth, C shows a promethazine, and D shows a pseudomonas brassicae fermentation broth compounded promethazine;
FIG. 3 shows an in-Pepper disease dish bacteriostasis test; wherein A shows a control, B shows a Pseudomonas fluorescens fermentation broth, C shows a Shenqimycin, and D shows a Pseudomonas fluorescens fermentation broth compounded with the Shenqimycin;
FIG. 4 shows an in-dish bacteriostasis test for tomato gray mold; wherein A shows a control, B shows Pseudomonas aeruginosa fermentation broth, C shows promethazine, and D shows Pseudomonas aeruginosa fermentation broth compounded promethazine.
Detailed Description
The invention discloses a composition and application thereof, and a person skilled in the art can use the content of the composition to appropriately improve the technological parameters. It is expressly noted that all such similar substitutions and modifications will be apparent to those skilled in the art, and are deemed to be included in the present invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those skilled in the relevant art that variations and modifications can be made in the methods and applications described herein, and in the practice and application of the techniques of this invention, without departing from the spirit or scope of the invention.
The invention prepares the synergistic composition by compounding the pseudomonas and the shenzinomycin.
The invention provides a decrement compounding method of pseudomonas and shenzinomycin, which is used for solving the problems in the background technology and applying the decrement compounding method to the prevention and treatment of various soil-borne diseases.
The pseudomonas agent comprises one or more of pseudomonas brassicae, pseudomonas fluorescens and pseudomonas viridis.
The pseudomonas, pseudomonas aeruginosa is preserved in China general microbiological culture collection center (CGMCC) on the 11 th month 02 of 2012, and the preservation number is CGMCC No.6774; pseudomonas brassicae is preserved in China general microbiological culture Collection center (CGMCC) in the year 2012, 11 and 02, and the preservation number is CGMCC No.6773; pseudomonas fluorescens is the same as M18 in China patent CN00119857.2 and is preserved in China general microbiological culture Collection center with a preservation number of CGMCC No.0462.
The types of the soil-borne disease prevention and control method provided by the invention comprise the following steps: wheat scab, rice sheath blight, pepper blight, watermelon fusarium wilt, cucumber gray mold, apple ring rot, tomato fusarium wilt, corn sheath blight, cotton fusarium wilt, melon gummy stem blight, potato late blight, and tomato gray mold.
The application method of the pseudomonas and the shenzinomycin after being compounded comprises soil treatment, leaf surface spraying, flushing, root irrigation, root dipping and the like.
Unless otherwise specified, the raw materials, reagents, consumables and instruments involved in the present invention are all commercially available and commercially available.
The invention is further illustrated by the following examples:
example 1: preparation of pseudomonas rape inoculant
The preparation method of the pseudomonas brassicae microbial inoculum comprises the following steps:
(1) Seed liquid preparation: inoculating pseudomonas brassicae into KB culture medium, and culturing for 14-16 h at the temperature of 28-30 ℃ and at the speed of 180-200 r/min to prepare seed liquid;
(2) Preparation of fermentation medium: taking 3% of sucrose, 1% of yeast powder, 0.5% of potassium chloride and the balance of water, adjusting pH=7 to prepare a fermentation medium, and sterilizing at 121 ℃ under high temperature and high pressure for 20min.
(3) Liquid fermentation of pseudomonas rape: and (2) sucking the seed liquid prepared in the step (1), inoculating the seed liquid into a fermentation culture medium according to the addition amount of 1%, and culturing for 24-30 h at the temperature of 28-30 ℃ and the rotating speed of 180-200 r/min.
(4) And collecting fermentation liquor, and uniformly mixing the fermentation liquor with diatomite and a protective agent (1% sodium alginate) according to a ratio of 10:3:1 to obtain the pseudomonas brassicae microbial agent.
Example 2: preparation of pseudomonas aeruginosa bacterial agent
The preparation method of the pseudomonas aeruginosa bacterial agent comprises the following steps:
(1) Seed liquid preparation: inoculating pseudomonas aeruginosa into KB culture medium, and culturing for 14-16 hours at the temperature of 28-30 ℃ and at the speed of 180-200 r/min to prepare seed liquid;
(2) Preparation of fermentation medium: taking yeast extract 0.5%, peptone 1%, sodium chloride 0.5%, na 2 HPO 4 0.02％、KH 2 PO 4 0.03%, anhydrous magnesium sulfate 0.1%, and water in balance, pH=7, and sterilizing at 121deg.C under high temperature and high pressure for 20min.
(3) Liquid fermentation of pseudomonas aeruginosa: and (2) sucking the seed liquid prepared in the step (1), inoculating the seed liquid into a fermentation culture medium according to the addition amount of 1%, and culturing for 24-30 h at the temperature of 28-30 ℃ and the rotating speed of 180-200 r/min.
(4) And collecting fermentation liquor, and uniformly mixing the fermentation liquor with diatomite and a protective agent (1% sodium alginate) according to a ratio of 9:2:1 to obtain the pseudomonas aeruginosa agent.
Example 3: preparation of Pseudomonas fluorescens bacterial agent
The preparation method of the Pseudomonas fluorescens bacterial agent comprises the following steps:
(1) Seed liquid preparation: inoculating Pseudomonas fluorescens into KB culture medium, and culturing for 14-16 h at the temperature of 28-30 ℃ and at the speed of 180-200 r/min to prepare seed liquid;
(2) Preparation of fermentation medium: taking corn flour 2%, bean cake flour 4%, peptone 1.5%, K 2 HPO 4 0.02％、KH 2 PO 4 0.03%, anhydrous magnesium sulfate 0.06%, glycerin 0.3%, and water in balance, pH=7, and sterilizing at 121deg.C under high temperature and high pressure for 20min.
(3) Pseudomonas fluorescens liquid fermentation: and (2) sucking the seed liquid prepared in the step (1), inoculating the seed liquid into a fermentation culture medium according to the addition amount of 1%, and culturing for 24-30 h at the temperature of 35-37 ℃ and the rotating speed of 180-200 r/min.
(4) And collecting fermentation liquor, and uniformly mixing the fermentation liquor with diatomite and a protective agent (1% sodium alginate) according to a ratio of 10:2:1 to obtain the pseudomonas fluorescens bacterial agent.
Example 4: preparation of Pseudomonas brassicae and shenqimycin (5:1) compound biological pesticide
The preparation of the compound biopesticide of pseudomonas brassicae and promethazine (5:1) comprises the following steps:
taking and fully mixing the pseudomonas brassicae microbial inoculum prepared in the example 1 with 1% of promethazine according to the proportion of 5:1 (v/v) to obtain a compound biological pesticide; and taking the compound biological pesticide to determine the colony concentration of pseudomonas brassicae, and determining the concentration to be 160 hundred million CFU/mL through a dilution coating.
Example 5: preparation of Pseudomonas brassicae and shenqimycin (1:1) compound biological pesticide
The preparation of the compound biopesticide of pseudomonas brassicae and promethazine (1:1) comprises the following steps:
the rape pseudomonas bacterial agent prepared in the example 1 and 1% of shenqimycin are fully and uniformly mixed according to the proportion of 1:1 (v/v) to obtain the compound biological pesticide. And taking the compound biological pesticide to determine the colony concentration of pseudomonas brassicae, and determining the concentration to be 100 hundred million CFU/mL through dilution coating.
Example 6: preparation of Pseudomonas fluorescens and shenqimycin compound biological pesticide
The preparation of the Pseudomonas fluorescens and shenqimycin compound biological pesticide comprises the following steps:
taking and fully mixing the Pseudomonas fluorescens bacterial agent prepared in the embodiment 3 with 1% of shenqimycin according to the proportion of 19:1 (v/v) to obtain a compound biological pesticide; the concentration of the pseudomonas fluorescens is 140 hundred million CFU/mL as measured by the dilution coating of the compound biological pesticide.
Example 7: preparation of pseudomonas aeruginosa and shenqinmycin compound biological pesticide
The preparation of the pseudomonas aeruginosa and shenqinmycin compound biological pesticide comprises the following steps:
taking the pseudomonas aeruginosa bacterial agent prepared in the example 2 and 1% of shenzinomycin, and fully and uniformly mixing according to the proportion of 10:1 (v/v) to obtain the compound biological pesticide. And taking the compound biological pesticide to determine the colony concentration of pseudomonas aeruginosa, and determining the concentration to be 130 hundred million CFU/mL through dilution coating.
Example 8: preparation of pathogen spore suspensions
Preparation of pathogenic bacteria spore suspension: activating pathogenic bacteria of watermelon wilt, botrytis cinerea and pepper epidemic disease on PDA plate, punching into bacterial blocks with diameter of 0.7mm with puncher, inoculating on PDA liquid culture medium, culturing at 28deg.C and 200r/min for 7d, filtering with gauze to remove mycelium to obtain spore suspension, counting with blood cell counting plate under microscope, and diluting with sterile water to 1×10 4 Individual spores/mL. The inoculation method of pathogenic bacteria is that the seedlings are slowly grown for 3d after the plant is transplanted, and 20mL of spore suspension liquid is irrigated into roots for each pot.
The rice sheath blight disease is caused by activating pathogenic bacteria on PDA plate and grafting bacterial blocks with uniform size on leaf sheath.
Effect example 1: prevention and treatment effect of compound promethazine and pseudomonas brassicae on watermelon fusarium wilt
After transplanting two leaves of the watermelon at one heart, inoculating pathogenic bacteria (the preparation method is as in example 8), applying the leaves to soil through root irrigation treatment after the leaves begin to generate slight yellowing symptoms, investigation on disease conditions after 14d, recording the numbers of plants of 0, 1, 2, 3 and 4 grades, and calculating disease indexes and disease prevention effects.
Potted plant experiment treatment for preventing and treating watermelon fusarium wilt: XG-CK0 (control group, clear water treatment without medicament), XG-CK1 (control group, 500 times diluted by 1% of shenzinomycin), XG-CK2 (control group, 500 times diluted by pseudomonas brassicae bacterial agent prepared in example 1), XG-T (treatment group, 500 times diluted by compound biopesticide prepared in example 4, namely 500 times diluted by pseudomonas brassicae bacterial agent mixed with 1% of shenzinomycin 5:1), wherein the shenzinomycin is provided by Shanghai agricultural products Co., ltd.
Grading watermelon fusarium wilt diseases: level 0: no disorder; stage 1:1 or 2 pieces She Qingwei yellow but grow normally; 2 stages: 1-2 leaves yellow or 1 cotyledon shows necrotic spots, and the true leaves slightly yellow and slightly wilt; 3 stages: all cotyledons die, true leaves obviously wilt and grow hindered; 4 stages: the whole plant seriously wilts or dies.
Incidence (%) =number of disease plants/total number of investigation×100
Disease index = Σ (number of disease stages x number of plants of the disease stage)/(4 x total number of plants) ×100
Control effect (%) = (control disease index-treatment disease index)/control disease index×100
The results are shown in fig. 1 and table 1, and the control effect of the compound biopesticide of the embodiment is remarkably improved.
Table 1: prevention and treatment effect of compound promethazine and pseudomonas brassicae on watermelon fusarium wilt
| Process numbering | Morbidity/% | Index of disease condition | Control effect/% |
| XG-CK0 | 80.0 a | 31.7 a | - |
| XG-CK1 | 28.9 c | 7.8 c | 75.4 b |
| XG-CK2 | 60.0 b | 22.2 b | 30.0 c |
| XG-T | 20.0 c | 5.0 c | 84.2 a |
Note that: XG is a watermelon abbreviation, and the lower case letters in the tables indicate that the difference is significant at the 5% level, as follows.
Effect example 2: prevention and treatment effect of compound promethazine and pseudomonas brassicae on rice sheath blight disease
The rice is transplanted and inoculated with pathogenic bacteria (the preparation method is as in example 8), the medicine is applied to the middle and lower parts of the rice by foliar spray after disease occurrence, investigation on disease occurrence condition is carried out after 14 days, the numbers of plants of 0, 1, 3, 5, 7 and 9 grades are recorded, and disease index and disease prevention effect are calculated.
Potted plant experiment for controlling rice sheath blight disease: SD-CK0 (control group, clear water treatment without medicament), SD-CK1 (control group, 500 times diluted by 1% of shenzinomycin), SD-CK2 (control group, 500 times diluted by pseudomonas brassicae bacterial agent prepared in example 1), SD-T (treatment group, 500 times diluted by compound biological pesticide prepared in example 5, namely 500 times diluted by pseudomonas brassicae bacterial agent and 1:1 (v/v) of shenzinomycin) are mixed
Grading rice sheath blight diseases: grade 0, no disease of whole plant; stage 1, 4 th leaf and leaf sheaths and leaf attacks below (sword leaf is 1 st leaf); 3 rd stage, 3 rd leaf and leaf sheath and leaf below; 5 th stage, 2 nd leaf and leaf sheath below it, leaf onset; stage 7, sword leaf blade and leaf sheath below it, blade onset; grade 9, onset of whole plant and early death.
Incidence (%) =number of disease plants/total number of investigation×100
Disease index = Σ (number of disease stages x number of plants of the disease stage)/(9 x total number of plants) x 100
Control effect (%) = (control disease index-treatment disease index)/control disease index×100
The results are shown in fig. 2 and table 2, and the control effect of the compound biopesticide of the embodiment is remarkably improved.
Table 2: prevention and treatment effect of compound promethazine and pseudomonas brassicae on rice sheath blight disease
| Treatment of | Morbidity/% | Index of disease condition | Control effect/% |
| SD-CK0 | 80.0 a | 20.7 a | - |
| SD-CK1 | 13.3 c | 3.2 c | 84.5 b |
| SD-CK2 | 40.0 b | 7.9 b | 61.8 c |
| SD-T | 11.1 c | 1.2 c | 94.2 a |
Note that: SD is the abbreviation of rice.
Effect example 3: application of shenqinmycin and fluorescent pseudomonas in preventing and treating pepper epidemic disease
Potted plant experiment for preventing and treating pepper epidemic disease: LJ-CK0 (control group, clear water treatment without medicament), LJ-CK1 (control group, 1% of shenqimycin is diluted 500 times), LJ-CK2 (control group, fluorescent pseudomonas as prepared in example 3 is diluted 500 times), LJ-T (treatment group, compound biological pesticide as prepared in example 6 is diluted 500 times, namely, the fluorescent pseudomonas as mixed with 1% of shenqimycin 19:1 (v/v) is diluted 500 times)
After 3 leaves of the capsicum are transplanted, pathogenic bacteria are inoculated (the preparation method is as in example 8), the capsicum is applied to soil through root irrigation treatment after the leaf wilting symptoms appear, disease condition investigation is carried out after 14 days, the numbers of plants of 0, 1, 2, 3, 4 and 5 grades are recorded, and disease indexes and disease prevention effects are calculated.
Classifying pepper epidemic disease: level 0: no disease; stage 1: the root and stem parts of the seedlings are slightly blackened, and leaves are not wilted or restorative wilting; 2 stages: the root and stem parts of the seedlings are blackened by 1-2 cm, the leaves are irrecoverable and wilt, and the leaves at the lower part are occasionally fallen off; 3 stages: the root and stem parts of the seedlings are blackened by more than 2cm, and the leaves are obviously wilted or fallen leaves are obviously removed; 4 stages: the root and stem parts of the seedlings become black and constrict, and all fallen leaves or plants except growing points wilt; 5 stages: plants die.
Incidence (%) =number of disease plants/total number of investigation×100
Disease index = Σ (number of disease stages x number of plants of the disease stage)/(5 x total number of plants) x 100
Control effect (%) = (control disease index-treatment disease index)/control disease index×100
The results are shown in fig. 3 and table 3, and the control effect of the compound biopesticide of the embodiment is remarkably improved.
Table 3: application of shenqinmycin and fluorescent pseudomonas in preventing and treating pepper epidemic disease
| Treatment of | Morbidity/% | Index of disease condition | Control effect/% |
| LJ-CK0 | 55.6 a | 20.0 a | - |
| LJ-CK1 | 17.8 c | 5.3 c | 73.5 b |
| LJ-CK2 | 28.9 b | 8.4 b | 58.0 c |
| LJ-T | 8.9 d | 2.0 d | 90.0 a |
Note that: LJ is an abbreviation for capsicum.
Effect example 4: compound effect of promethazine and pseudomonas aeruginosa on preventing and treating gray mold of tomatoes
Potting experiment for preventing and treating gray mold of tomatoes: FQ-CK0 (control group, clear water treatment without medicament), FQ-CK1 (control group, 500 times diluted by 1% of shenqimycin), FQ-CK2 (control group, 500 times diluted by pseudomonas aeruginosa agent prepared in example 2), FQ-T (treatment group, 500 times diluted by compound biological pesticide prepared in example 7, namely 500 times diluted by pseudomonas aeruginosa agent mixed with 1% of shenqimycin 10:1 (v/v))
The 3 leaves of the tomato are transplanted and inoculated with pathogenic bacteria (the preparation method is as in example 8), after the single leaf has the disease spots, the single leaf is applied into soil through root irrigation treatment, disease condition investigation is carried out after 14 days, the numbers of plants of 0, 1, 3, 5, 7 and 9 grades are recorded, and the disease index and the disease prevention effect are calculated.
Grading tomato gray mold diseases: level 0: no disease spots; stage 1: 3 disease spots are arranged on a single leaf; 3 stages: 4-6 disease spots are arranged on a single blade; 5 stages: 7-10 disease spots are arranged on a single blade; 7 stages: the single leaf has 11-20 disease spots, and part of the single leaf is densely packed into a sheet; stage 9: the single leaf has more than one quarter of densely-occupied disease spots.
Incidence (%) =number of disease plants/total number of investigation×100
Disease index = Σ (number of disease stages x number of plants of the disease stage)/(9 x total number of plants) x 100
Control effect (%) = (control group disease index-treatment group disease index)/control disease index x 100.
The results are shown in fig. 4 and table 4, and the control effect of the compound biopesticide of the embodiment is remarkably improved.
Table 4: compound effect of promethazine and pseudomonas aeruginosa on preventing and treating gray mold of tomatoes
| Treatment of | Morbidity/% | Index of disease condition | Control effect/% |
| FQ-CK0 | 60.0 a | 16.5 a | - |
| FQ-CK1 | 17.8 c | 4.0 c | 75.8 b |
| FQ-CK2 | 35.6 b | 6.4 b | 61.2 c |
| FQ-T | 8.9 d | 1.0 c | 93.9 a |
Note that: FQ is the abbreviation for tomato.
As can be seen from the potting experimental results of examples 5, 6, 7 and 8, after the shenqimycin and the pseudomonas brassicae, the pseudomonas aeruginosa and the pseudomonas fluorescens are compounded, compared with the single application of the shenqimycin, the pseudomonas brassicae, the pseudomonas viridis and the pseudomonas fluorescens respectively, the disease condition of potted crops is obviously reduced, and the control effect on watermelon fusarium wilt, rice sheath blight, pepper epidemic disease and tomato gray mold is improved.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. A composition comprising pseudomonas and promethazine.
2. The composition of claim 1, wherein the volume ratio of said pseudomonas to said shenzinomycin is from (1 to 19): 1.
3. The composition of claim 1 or 2, wherein the pseudomonas comprises one or more of pseudomonas brassicae, pseudomonas fluorescens, or pseudomonas aeruginosa.
4. A composition according to claim 3, comprising:
the preservation number of the pseudomonas brassicae is CGMCC No.6774; and/or
The preservation number of the pseudomonas fluorescens is CGMCC No.0462; and/or
The preservation number of the pseudomonas aeruginosa is CGMCC No.6773.
5. A composition according to claim 3 or 4 or claim 4, comprising:
the viable count of the pseudomonas brassicae is 100 hundred million CFU/mL to 190 hundred million CFU/mL; and/or
The viable count of the pseudomonas fluorescens is 75 hundred million CFU/mL to 140 hundred million CFU/mL; and/or
The living bacteria number of the pseudomonas aeruginosa is 75 hundred million CFU/mL to 140 hundred million CFU/mL.
6. The composition of any one of claims 1 to 5, wherein said shenzinomycin is used at a concentration of 1% (v/v).
7. Use of a composition according to any one of claims 1 to 6 for controlling soil-borne diseases;
the soil-borne disease comprises one or more of wheat scab, rice sheath blight, pepper blight, watermelon fusarium wilt, cucumber gray mold, apple ring rot, tomato fusarium wilt, corn sheath blight, cotton fusarium wilt, melon gummy stem blight, potato late blight or tomato gray mold.
8. Use of a composition according to any one of claims 1 to 6 for the preparation of a pesticide for controlling soil-borne diseases;
the soil-borne disease comprises one or more of wheat scab, rice sheath blight, pepper blight, watermelon fusarium wilt, cucumber gray mold, apple ring rot, tomato fusarium wilt, corn sheath blight, cotton fusarium wilt, melon gummy stem blight, potato late blight or tomato gray mold.
9. Pesticide, characterized by comprising a composition according to any one of claims 1 to 6, together with acceptable adjuvants or auxiliaries.
10. A method for the preparation of a pesticide according to claim 9, comprising diluting the composition according to any one of claims 1 to 6 by a factor of 500 to obtain the pesticide.
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Citations (3)
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|---|---|---|---|---|
| US6348193B1 (en) * | 1999-09-02 | 2002-02-19 | Board Of Trustees Of Michigan State University | Method for inhibiting pathogenic fungi and bacteria in plants using Pseudomonas aureofaciens |
| CN104017744A (en) * | 2013-11-07 | 2014-09-03 | 上海交通大学 | Preparation method and application of pseudomonas chlororaphis for resisting disease and promoting growth |
| CN114921364A (en) * | 2022-04-26 | 2022-08-19 | 慕恩（广州）生物科技有限公司 | Pseudomonas brassicae, biochemical fungicide and application thereof |
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| US6348193B1 (en) * | 1999-09-02 | 2002-02-19 | Board Of Trustees Of Michigan State University | Method for inhibiting pathogenic fungi and bacteria in plants using Pseudomonas aureofaciens |
| CN104017744A (en) * | 2013-11-07 | 2014-09-03 | 上海交通大学 | Preparation method and application of pseudomonas chlororaphis for resisting disease and promoting growth |
| CN114921364A (en) * | 2022-04-26 | 2022-08-19 | 慕恩（广州）生物科技有限公司 | Pseudomonas brassicae, biochemical fungicide and application thereof |
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