CN109497077B - Application of corybuterol B in preparation of bacterial or fungal inhibitor - Google Patents
Application of corybuterol B in preparation of bacterial or fungal inhibitor Download PDFInfo
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- CN109497077B CN109497077B CN201811421548.8A CN201811421548A CN109497077B CN 109497077 B CN109497077 B CN 109497077B CN 201811421548 A CN201811421548 A CN 201811421548A CN 109497077 B CN109497077 B CN 109497077B
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- 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
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
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Abstract
The invention discloses an application of Corbuterol B in preparation of a bacterial or fungal inhibitor, and the applicant finds that Corbuterol B (Kobutomycin B) has an inhibiting effect on various bacteria and fungi for the first time, wherein the bacteria comprise ralstonia solanacearum, xanthomonas oryzae, pseudomonas syringae, staphylococcus aureus and xanthomonas citri; the fungi include Fusarium solani, Fusarium oxysporum, Verticillium gossypii, Magnaporthe grisea, Botrytis cinerea, Gibberella tritici, Sclerotinia sclerotiorum, and Penicillium citricola. The substance has strong antagonism especially on the corybuterol B, and provides selectivity for the preparation of biological preparations for preventing and treating bacterial wilt.
Description
Technical Field
The invention belongs to the technical field of agricultural microbiology, and particularly relates to an application of corybuterol B in preparation of a bacterial or fungal inhibitor.
Background
Bacterial wilt (Bacterial wilt) is a devastating soil-borne disease in the world, is the second place of ten major Bacterial diseases in the world, and is called as 'cancer' of plants due to rapid onset and lack of economic and effective prevention and treatment measures. The pathogen is Ralstonia solanacearum (Ralstonia solanacearum for short), which generally invades from the root or wound of a plant and propagates in the vascular bundle in large quantities, so that the vascular bundle is blocked and necrotized, and the plant is withered and dies due to water shortage. According to statistics, the bacterium can infect more than 300 plants of 45 families, and solanaceae crops including tomatoes, eggplants, hot peppers, tobaccos, potatoes and the like are the most seriously damaged, the death rate of seriously ill fields reaches more than 90 percent, and destructive loss is brought to agricultural production.
The control method of solanaceae bacterial wilt comprises chemical control, agricultural control, soil disinfection, biological control and the like. (1) Chemical control: also known as pesticide control, is the most common method for controlling plant diseases. Currently, the most common chemical agent for controlling bacterial wilt is agricultural streptomycin, which has been completely banned by Ministry of agriculture as a pesticide due to its residue, toxicity, environmental safety and other problems. (2) Agricultural control: the paddy-upland rotation, rotation and the like are adopted, the cultivation system or cultivation mode is changed, the continuous cropping of crops is avoided, and the bacteria content in soil is reduced. (3) Soil disinfection: the soil is disinfected by chemical agents to thoroughly kill pathogenic bacteria in the soil, wherein the disinfection effect of methyl bromide is the best, but the methyl bromide is forbidden globally due to the damage of the methyl bromide to the ozone layer. (4) Biological control: the biological control has the characteristics of good environmental compatibility, high safety and the like, and is a key point in solanaceae bacterial wilt control research.
Cobuterol B (Kobutamycin B) is a microbial antibiotic, and has antagonistic action on Bacillus subtilis, Escherichia coli, yeast and the like. However, there is no report on the inhibitory effect of coxsackienin B on Ralstonia solanacearum.
Disclosure of Invention
The invention provides an application of corynebacterium sp in preparing a bacterial or fungal inhibitor, wherein the bacteria comprise ralstonia solanacearum, xanthomonas oryzae, pseudomonas syringae, staphylococcus aureus and xanthomonas citri; the fungi comprise fusarium solani, fusarium oxysporum, verticillium dahliae, rice blast fungi, cucumber botrytis cinerea, wheat gibberella, sclerotinia sclerotiorum and penicillium citrinum; the corynebacterium sp has good activity of inhibiting ralstonia solanacearum, and provides selectivity for preparing biological preparation for preventing and treating bacterial wilt.
In order to achieve the purpose, the invention adopts the following technical measures:
the chrysotidin B (5-Methyl-7- (2 '-methylbutylyloxy) -3' -acetoxy) butyllidene-1 a,2,3,7-tetrahydrocyclopent (B) oxireno (C) pyridine) has the molecular weight of 361.43, and the molecular formula is C20H27NO5The chemical formula is as follows:
the application of the corynebacterium sp in preparing the bacterial or fungal inhibitor comprises the steps of using the corynebacterium sp as a unique effective component or one of the effective components for preparing the bacterial or fungal inhibitor, wherein the bacteria comprise ralstonia solanacearum, xanthomonas oryzae, pseudomonas syringae, staphylococcus aureus and xanthomonas citri; the fungi include fusarium solani, fusarium oxysporum, verticillium dahliae, rice blast, cucumber botrytis cinerea, wheat gibberella, sclerotinia sclerotiorum and penicillium citrinum.
The use of corybuterol B in the manufacture of a medicament for the treatment or prevention of a disease caused by a bacterial disease or a fungal disease; the bacterial diseases comprise xanthomonas oryzae, pseudomonas syringae, staphylococcus aureus, xanthomonas citri and ralstonia solanacearum; the fungi include fusarium solani, fusarium oxysporum, verticillium dahliae, rice blast, cucumber botrytis cinerea, wheat gibberella, sclerotinia sclerotiorum and penicillium citrinum.
Compared with the prior art, the invention has the following advantages:
firstly, the method comprises the following steps: the invention discovers that the solanaceous plant strain has the activity of resisting solanaceous bacteria for the first time;
secondly, the method comprises the following steps: the compound has strong antagonistic capability to ralstonia solanacearum, and the minimum inhibitory concentration is only 14.2 mu g/ml; provides new selectivity for the preparation of novel biological control preparation for the ralstonia solanacearum disease.
Drawings
FIG. 1 is a bacteriostatic diagram of corybuterol B against ralstonia solanacearum compounds.
Detailed Description
The experimental procedures in the following examples are reported as conventional procedures in microbiology unless otherwise specified. Either commercially available or synthetic coxtiramicin B may complete the invention. The corybuterol B used in the embodiment of the invention is synthesized according to a disclosed conventional mode, and has the following chemical formula:
example 1:
determination of minimum inhibitory concentration of Corbuterol B against ralstonia solanacearum by using tube-disc method
The principle is as follows: it is deduced from the diffusion law that the logarithmic value of the total amount of antibiotic is linear with the square of the radius of inhibition. When the concentration of the antibiotic reaches or exceeds the MIC value, the test bacteria are inhibited and cannot reproduce, so that a transparent inhibition zone is presented.
LogY＝kX2+ b (Y is concentration, X is radius of inhibition zone)
1) The ralstonia solanacearum is streaked and activated to obtain a single colony, and the single colony is selected to be inoculated into an SPA culture medium at 28 ℃ and cultured at 150rpm until the logarithmic phase.
2) Cobuterol B was formulated at concentrations of 7.1,14.2, 28.4,56.8, 113.6. mu.g/mL, with water as a negative control.
3) Adding 1% of pathogenic bacteria into a melted solid culture medium (added when a shake flask is not too hot), placing an Oxford cup (with the inner diameter of 6mm, the outer diameter of 7.8mm and the height of 10mm, the same below) in a poured flat plate, adding 100ul of coelutin B solution with different concentrations, culturing in a incubator at 28 ℃, and observing the result.
Using the above experiment, the equation obtained is LogY ═ 0.0005X2+1.1739, the minimum inhibitory concentration was found to be 14.2. mu.g/ml.
Experimental example 2:
determining the activity of the corybuterol B against common pathogenic bacteria by a tube-disc method:
1) after the pathogenic bacteria Xanthomonas oryzae (Xanthomonas oryzae), Pseudomonas syringae (Pseudomonas syringae) and Xanthomonas citri (Xanthomonas citri) were activated, a single colony was selected and inoculated into LB medium at 28 ℃ and 150rp m to be cultured until logarithmic phase. The pathogenic bacterium Staphylococcus aureus (Staphylococcus aureus) was cultured at 28 ℃ and 150rpm to logarithmic growth phase.
2) Cobuterol B was prepared at a concentration of 100. mu.g/mL, with water as a negative control.
3) 1% of pathogenic bacteria was added to a melted solid medium (1ml of bacterial solution +99ml of medium) (added while shaking hands free), an Oxford cup was placed in a poured plate and 100ul of brevethin B was added, 3 replicates were performed, Staphylococcus aureus was cultured at 37 ℃ and Rhizoctonia solani, Rhizopus cucumerinum and Leptosphaera citri were cultured at 28 ℃, and the results are shown in Table 1.
Table 1: antibacterial spectrum of chrysotilin B
+: shows an effect, -: indicating no effect
Experimental example 3:
method for measuring activity of coxiticin B against common pathogenic fungi by perforation method
1) Pathogenic fungi, Fusarium solani (Fusarium oxysporum), Fusarium oxysporum (Fusarium oxysporum), Verticillium dahliae (Verticillium dahliae), pyricularia oryzae (magnaporthe), botrytis cinerea (botrytis cinerea), Fusarium graminearum (Fusarium graminearum), sclerotinia sclerotiorum (sclerotinia sclerotiorum) and Penicillium citrinum (Penicillium italicum), are activated, and hypha is selected to be inoculated into a PDA culture medium for culture at 28 ℃.
2) Cobuterol B was prepared at a concentration of 100. mu.g/mL, with water as a negative control.
3) The pellet was picked from the center of a PDA solid medium plate, 6 wells (the diameter of the well was 4mm) were punched with a punch at equal intervals, 20. mu.l of Corbutomycin B was added thereto, and the mixture was cultured at 28 ℃ in 3 replicates, and the results of observation are shown in Table 2.
Table 2: cobuterol B antifungal spectra
+: shows an effect, -: indicating no effect.
Claims (2)
1. The application of the corybuterol B in preparing a bacterial or fungal inhibitor is as follows: ralstonia solanacearum, xanthomonas oryzae, pseudomonas syringae or xanthomonas citri; the fungus is fusarium solani, fusarium oxysporum, cucumber botrytis cinerea or penicillium citrinum.
2. The application of the corybuterol B in preparing the medicine for treating or preventing diseases caused by bacterial diseases or fungi, wherein the bacterial diseases are Xanthomonas oryzae, Pseudomonas syringae, Xanthomonas citri or ralstonia solanacearum; the fungus is fusarium solani, fusarium oxysporum, cucumber botrytis cinerea or penicillium citrinum.
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| WO2015017866A1 (en) * | 2013-08-02 | 2015-02-05 | Enevolv, Inc. | Processes and host cells for genome, pathway, and biomolecular engineering |
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| "KOBUTIMYCINS A AND B, NEW ALKALOID ANTIBIOTICS PRODUCED BY A Streptomyces STRAIN";KENJI KANBE等;《The Journal of Antibiotics》;19921231;第45卷(第10期);第1702页表3 * |
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