A bactericidal composition containing bromothionil and oxytetracycline hydrochloride
By using bactericidal compositions of bromocrinone and oleycin hydrochloride, the problem of prevention and treatment of corn white spots was solved, and effective prevention and treatment of corn white spots was achieved, and the yield and quality of corn were significantly improved.
Patent Information
- Application Number
- CN202311053998.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-08-21
- Publication Date
- 2024-12-13
- Estimated Expiration
- 2043-08-21
AI Technical Summary
The existing technology cannot effectively prevent and treat corn white spots, resulting in a significant reduction in corn yield and lack of effective pesticide measures.
Using a bactericidal composition containing bromocrin and olefinicidin hydrochloride, it is prepared by mixing 25% bromocrin and olefinicidin hydrochloride in a ratio of 1:4-4:1, combined with pesticide-enabled auxiliary materials. Wettable powder or water agent, spray on the diseased leaves to prevent and treat corn white spots.
This composition can effectively inhibit the growth and spread of pathogenic bacteria of white spot disease in corn, significantly reduce the impact of the disease on corn yield and quality, and has simple preparation and application methods, and has great promotion and application value.
Smart Images
Figure CN117296847B_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of pesticide bacterial control, and particularly relates to a bactericidal composition containing bromothionil and oxytetracycline hydrochloride. Background Art
[0002] Corn white spot is a bacterial disease of corn caused by infection by Pantoe bacteria. The disease usually begins to occur during the heading and flowering stage of corn, breaks out during the grain filling stage, and progresses rapidly. The lesions begin to infect the old leaves at the base and gradually spread upward. They are initially small, light green, and scattered on the surface of the leaves. As time goes by, the lesions turn white, dry, have brown edges, and are densely dispersed on the surface of the leaves. They usually appear in irregular shapes such as nearly circular, oval, and square, and are usually 3.0-20.0mm in diameter. The lesions are penetrating, that is, the size and color of the lesions on the front and back of the leaves are exactly the same or almost the same. The edges of the lesions are neat and clear, without radial or unclear extension lines or extended spots, and the lesions have symptoms of sudden downward movement. In the later stage, ruptures and perforations will appear. In the later stage of the disease, multiple lesions will further expand and merge into irregular large lesions, causing the death of the entire leaf. The lesions continue to develop and become larger and larger, until the entire corn plant falls over and withers, greatly affecting the corn yield, causing a serious shortening of the corn growth period, an average yield reduction of about 70%-85%, and severe plots may even result in a total loss of grains.
[0003] At present, large-scale outbreaks of corn white spot disease have been monitored in many provinces and cities in my country, and they are showing an increasingly rapid radiation and spread trend. In 2019, the first report of the disease occurred only in Gengma County, Lincang City, Wenshan City and other individual areas in Yunnan; but by 2020, the disease had covered the entire Yunnan Province; in 2021, the disease had spread to Guangxi, Sichuan, Guizhou, Chongqing, Hubei, Shaanxi and other places, causing a huge impact on my country's corn production. Moreover, even if calculated at the current rate of spread, white spot disease can cover all corn-growing areas in China within 3-5 years. Moreover, since the disease is a new disease and there is a lack of experience accumulation, there is currently no special pesticide for the prevention and treatment of corn white spot disease. After the disease occurs, it can only rely on the stress resistance of corn itself to resist the course of corn white spot disease. At present, propiconazole, myclobutanil, fenpropimorph and other agents are used for prevention and control during the onset period, but their effects are limited.
[0004] At present, the research on corn white spot disease only stays in the relatively superficial research such as disease investigation or simple identification. For example, CN115058486A discloses a disease resistance identification method for corn white spot disease, which classifies the severity of the disease according to the symptoms of corn white spot disease leaf disease, calculates the disease index, and evaluates the disease resistance, which can help improve the screening efficiency of disease-resistant materials. CN114451295A discloses a corn hybrid breeding method with high resistance to the main diseases of white spot disease. By using fertile corn species with resistance to black shank disease and white spot disease as the original resource and self-pollinating, hybridizing and intercrossing the corn species, it can show high resistance to large spot disease, gray spot disease, white spot disease and head smut to a certain extent. However, there is still no effective method for corn prevention and control after onset, so a medicine that can effectively prevent and control corn white spot disease is urgently needed. Summary of the invention
[0005] In view of the above problems, the present invention provides a fungicide composition containing bromothionil and oxytetracycline hydrochloride, which is prepared by mixing 25% bromothionil and oxytetracycline hydrochloride with auxiliary materials allowed for use and acceptable to pesticides. The fungicide composition can effectively prevent and treat the pathogenic bacteria of corn white spot disease, thereby ensuring the yield and quality of corn.
[0006] To achieve the above purpose, the technical effects adopted by the present invention are as follows:
[0007] A bactericidal composition containing bromothionil and oxytetracycline hydrochloride, wherein the active ingredients are bromothionil and oxytetracycline hydrochloride, and the mass ratio of bromothionil to oxytetracycline hydrochloride is 1:4-4:1.
[0008] Preferably, the pharmaceutical composition consists of the following components in parts by weight: 20%-80% bromothionil, 20%-80% oxytetracycline hydrochloride, and the remainder is excipients permitted and acceptable in pesticide science.
[0009] Further preferably, the concentration of bromothionil is 20%-30%.
[0010] Further preferably, the excipients permitted and acceptable in pesticide science are solvents, dispersants, antifreeze agents, stabilizers, emulsifiers, wetting agents or other permitted adjuvants that help stabilize and exert the efficacy of the active ingredients.
[0011] Preferably, the dosage form of the pharmaceutical composition is any one of granules, water dispersible granules, wettable powder, water dispersible granules and suspension.
[0012] The present invention also provides an application of a bactericidal composition containing bromothionil and oxytetracycline hydrochloride in preventing bacterial diseases of crops, and the application method comprises the following steps:
[0013] (1) preparing bromothionil into a raw material with a concentration of 20%-30%;
[0014] (2) mixing the bromothionil raw material obtained in step (1) with oxytetracycline hydrochloride and pharmaceutical excipients permitted for use and acceptable in pesticide science in proportion to prepare a wettable powder;
[0015] (3) diluting the wettable powder with a small amount of water, then diluting it to 1000-2000 times, and mixing evenly to obtain an aqueous fungicide composition;
[0016] (4) Evenly spray the aqueous fungicide composition onto the leaves of crops suffering from bacterial diseases in the afternoon on a sunny day.
[0017] Preferably, the bacterial disease is white spot disease.
[0018] Preferably, the crop is any one of corn, sorghum, rice and wheat, preferably corn.
[0019] Preferably, the spraying cycle is every 14-20 days.
[0020] Beneficial Effects
[0021] 1. According to the characteristics of the pathogenic bacteria of corn white spot disease, two bactericidal substances that can inhibit the pathogenic bacteria of corn white spot disease were screened: bromothionil and oxytetracycline hydrochloride; then bromothionil and oxytetracycline hydrochloride were mixed to prepare a bactericidal composition, which can further inhibit the growth and proliferation of the pathogenic bacteria of corn white spot disease, effectively prevent and control the occurrence or outbreak of corn white spot disease, and greatly reduce the impact of white spot disease on corn yield and quality.
[0022] 2. The preparation method of the bactericidal composition is simple, the prevention and control effect is good, the application method is simple and easy to operate, and it has great promotion and application value. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 These are plants susceptible to corn white spot disease. In the picture, A is a photo of the leaves of diseased corn plants, and B is a large area of corn field withering after being infected.
[0024] Figure 2 The figure is a graph showing the inhibitory effects of different medicinal ingredients in Example 1 on the pathogen of corn white spot disease, wherein A is 25% bromothionil, B is oxytetracycline hydrochloride, C is streptomycin, D is copper hydroxide, E is Bordeaux mixture, F is cuprous oxide, G is agricultural streptomycin, H is 10% difenoconazole, and I is sterile water.
[0025] Figure 3The inhibitory effect diagram of the medicinal composition prepared in Examples 2-3 and Comparative Examples 1-3 on the pathogen of corn white spot disease, wherein A is Comparative Example 3 (sterile water), B is Comparative Example 2 (oxytetracycline hydrochloride), C is Comparative Example 1 (25% bromothionil), D is Example 1-1 (25% bromothionil: oxytetracycline hydrochloride = 1:1), E is Example 1-2 (25% bromothionil: oxytetracycline hydrochloride = 2:1), F is Example 1-3 (25% bromothionil: oxytetracycline hydrochloride = 4:1), G is Example 1-4 (25% bromothionil: oxytetracycline hydrochloride = 1:2), and H is Example 1-5 (25% bromothionil: oxytetracycline hydrochloride = 1:4).
[0026] Figure 4 The figures are the inhibitory effects of the medicinal compositions prepared in Examples 2-3 and Comparative Examples 1-2 on the pathogens of corn white spot disease at different times, in which A is the inhibitory effect of Comparative Example 2 (oxytetracycline hydrochloride) for 2 days, and a is the inhibitory effect of Comparative Example 2 (oxytetracycline hydrochloride) for 10 days; B is the inhibitory effect of Comparative Example 1 (25% bromothionil) for 2 days, and b is the inhibitory effect of Comparative Example 1 (25% bromothionil) for 10 days; C is the inhibitory effect of Example 1-1 (25% bromothionil: oxytetracycline hydrochloride = 1:1) for 2 days, and c is the inhibitory effect of Example 1-1 (25% bromothionil: oxytetracycline hydrochloride = 1:1) for 10 days.
[0027] Figure 5 This is the field application of the optimal ratio of the combination of pesticides for corn white spot disease in Example 4. In the figure, A and B are corn leaves in the early stage of the disease; Figure C is the diseased state of corn leaves without the use of combined pesticides for prevention and control; Figure D is the diseased state of corn leaves after the use of combined pesticides for prevention and control. DETAILED DESCRIPTION
[0028] The technical solution of the present invention is further explained below in conjunction with the accompanying drawings and specific embodiments. It is worth noting that the following embodiments are only preferred embodiments of the present invention and should not be construed as limiting the present invention. The protection scope of the present invention shall be subject to the contents recorded in the claims. Any modification or replacement made by a person skilled in the art to the technical solution of the present invention without creative work shall fall within the protection scope of the present invention.
[0029] Pathogen of corn white spot disease: The pathogen was obtained from diseased corn leaves according to Koch's postulates and sent to Wuhan BGI for sequencing. After the sequence was obtained, NCBI BLASTn analysis revealed that the pathogen was a new species of Pantoea bacteria. The bacteria were preserved in the laboratory of the Biotechnology Research Center of Three Gorges University.
[0030] Example 1 Screening of effective inhibitory components for corn white spot disease
[0031] Common pesticide ingredients on the market were selected to conduct inhibition experiments on corn white spot disease pathogens. The specific steps are as follows:
[0032] (1) Cultivate the pathogen of corn white spot disease to obtain a bacterial solution, and then dilute the bacterial solution to 3×10 8 CFU / mL, spread on LB plate medium and cultured overnight at 37°C;
[0033] (2) Common bacterial pesticides 25% bromothionil wettable powder, sinoprotinin wettable powder, copper hydroxide, Bordeaux mixture, cuprous oxide, agricultural streptomycin, 10% difenoconazole and oxytetracycline hydrochloride were diluted to 1 mg / mL with sterile water and then cut into 1 cm 2 The filter paper discs were immersed in it, and sterile water was used as a control;
[0034] (3) After 3 minutes, the filter paper disc was taken out and placed in the LB plate culture medium obtained in step (1), cultured at 37° C., and the growth of corn white spot disease was observed. After 24 hours, the antibacterial effect was observed.
[0035] The results are as follows Figure 2 As shown in the figure, different pesticide components have different inhibitory effects on the pathogen of corn white spot disease. Among them, 25% bromothionil and oxytetracycline hydrochloride have the best inhibitory effects on the pathogen of corn white spot disease. Biotin, copper hydroxide, Bordeaux mixture, cuprous oxide, agricultural streptomycin, and 10% difenoconazole have almost no inhibitory effect on the pathogen. Therefore, bromothionil and oxytetracycline hydrochloride were selected for further study.
[0036] Example 2 Preparation of pharmaceutical compositions with different ratios
[0037] (1) Preparation of 25% bromothionil: 25 parts of bromothionil, 5 parts of hexaconazole, 10 parts of xylene, 5 parts of DMF, 10 parts of emulsifier ME-6, 5 parts of n-butanol and 3 parts of ethylene glycol were taken, and then distilled water was added to make up to 100 parts, and the mixture was stirred and mixed to prepare 25% bromothionil;
[0038] (2) Take an appropriate amount of 25% bromothionil prepared in step (1), weigh oxytetracycline hydrochloride wettable powder, add water to dilute to 1000 times, and prepare a mixed aqueous solution. The mixing ratio of bromothionil and oxytetracycline hydrochloride is shown in Table 1:
[0039] Table 1 Medicinal compositions with different ratios
[0040] project 25% Bromothionil dosage Oxytetracycline Hydrochloride Dosage Bromothionil: Oxytetracycline Hydrochloride 1-1 30mL 30g 1:1 1-2 40mL 20g 2:1 1-3 48mL 12g 4:1 1-4 20mL 40g 1:2 1-5 12mL 48g 1:4
[0041] Example 3
[0042] The antibacterial experiment was carried out on the pharmaceutical composition with the ratio of 5 prepared in Example 1, and the specific steps were as follows:
[0043] (1) Cultivate the pathogen of corn white spot disease to obtain a bacterial solution, and then dilute the bacterial solution to 3×10 8CFU / mL, spread on LB plate medium and cultured overnight at 37°C;
[0044] (2) Cut some 1cm 2 Filter paper discs are respectively soaked in the 5 pharmaceutical compositions of the proportions prepared in Example 1;
[0045] (3) After soaking for 3 minutes, the filter paper disc was taken out and placed in the LB plate culture medium obtained in step (1), cultured at 37°C, and the growth of corn white spot disease was observed. After 24 hours, the diameter of the inhibition zone was measured and counted. The results are shown in Table 2.
[0046] Comparative Example 1
[0047] Take 60 mL of 25% bromothionil and dilute it 1000 times with water, then soak the filter paper disc. After 3 minutes, take out the filter paper disc containing 3×10 8 The samples were cultured on LB plates with pathogens at 37°C and 400 μg / mL. The diameters of the inhibition zones were measured and counted after 24 h. The results are shown in Table 2.
[0048] Comparative Example 2
[0049] Take 60g of oxytetracycline hydrochloride wettable powder and dilute it 1000 times with water, then soak the filter paper disc. After 3 minutes, take out the filter paper disc containing 3×10 8 The samples were cultured on LB plates with pathogens at 37°C and 400 μg / mL. The diameters of the inhibition zones were measured and counted after 24 h. The results are shown in Table 2.
[0050] Comparative Example 3
[0051] Soak the filter paper disc in sterile water for 3 min and take it out to the filter paper containing 3×10 8 The samples were cultured on LB plates with pathogens at 37°C and 400 μg / mL. The diameters of the inhibition zones were measured and counted after 24 h. The results are shown in Table 2.
[0052] Table 2 Antibacterial experimental data of different drugs
[0053]
[0054] From Table 2 and Figure 3 It can be seen that bromothionil and oxytetracycline hydrochloride have a good synergistic enhancement effect. When bromothionil and oxytetracycline hydrochloride wettable powder are mixed in different proportions, the inhibitory effect on the pathogen of corn white spot disease is more significant, the antibacterial time is more lasting, the incidence rate is reduced, and the disease prevention effect is improved.
[0055] When 25% bromothionil was used alone in Comparative Example 1 and oxytetracycline hydrochloride was used alone in Comparative Example 2 for bacteriostasis, after 10 days of use, the diameter of the inhibition zone was significantly reduced and accompanied by the regeneration of pathogenic bacteria. However, the bactericidal composition prepared by mixing 25% bromothionil and oxytetracycline hydrochloride in a mass ratio of 1:1 in Example 1 did not significantly reduce the diameter of the inhibition zone as the use time increased, indicating that the prepared bactericidal composition has long-lasting bacteriostasis and has an ideal bacteriostasis effect on the pathogenic bacteria of corn white spot disease. At the same time, it is shown that the prepared bactericidal composition can have a longer duration of efficacy when applied to corn plants suffering from white spot disease, which can help guide the frequency of use of the bactericidal composition.
[0056] Example 4
[0057] (1) Preparation of 25% bromothionil: 25 parts of bromothionil, 5 parts of hexaconazole, 10 parts of xylene, 5 parts of DMF, 10 parts of emulsifier ME-6, 5 parts of n-butanol and 3 parts of ethylene glycol were taken, and then distilled water was added to make up to 100 parts, and the mixture was stirred and mixed to prepare 25% bromothionil;
[0058] (2) 25% bromothionil and oxytetracycline hydrochloride were mixed in a ratio of 1:1, a small amount of water was first added to the wettable powder to dilute and dissolve, and then diluted with water to 2000 times to obtain a fungicide composition aqueous solution;
[0059] (4) Select corn plants at the early stage of disease susceptibility and the jointing stage, and evenly spray the aqueous fungicide composition prepared in step (3) onto the leaf surfaces of the corn plants on a sunny afternoon with no strong wind and no rainfall within 2 hours. Spray three times in succession with an interval of 14 days between each spraying. Observe the size of the lesions 3 days after the third spraying and record the growth of the corn plants.
[0060] The results are as follows Figure 5 It shows that the corn plants that were not sprayed with the fungicide composition had continuous expansion of lesions and stagnant leaf growth ( Figure 5 A and D); The corn plants sprayed with the fungicide aqueous solution did not have any further enlargement of the lesions, and the corn leaves could grow normally ( Figure 5 B and D), the infectiousness weakened and no large-scale outbreak occurred.
Claims
1. A bactericidal composition containing bromothionil and oxytetracycline hydrochloride, characterized in that: The active ingredients are bromothionil and oxytetracycline hydrochloride, and the mass ratio of bromothionil to oxytetracycline hydrochloride is 1:4-4:
1.
2. A bactericidal composition containing bromothionil and oxytetracycline hydrochloride according to claim 1, characterized in that: The bactericidal composition is composed of the following components in parts by weight: 20%-80% of bromothionil, 20%-80% of oxytetracycline hydrochloride, and the remainder is auxiliary materials permitted and acceptable in pesticide science.
3. A bactericidal composition containing bromothionil and oxytetracycline hydrochloride according to claim 1 or 2, characterized in that: The concentration of bromothionil is 20%-30%.
4. A bactericidal composition containing bromothionil and oxytetracycline hydrochloride according to claim 3, characterized in that: The excipients permitted and acceptable in pesticide science are solvents, dispersants, antifreeze agents, stabilizers, emulsifiers, wetting agents or other permitted adjuvants that help stabilize and exert the efficacy of the active ingredients.
5. A bactericidal composition containing bromothionil and oxytetracycline hydrochloride according to claim 4, characterized in that: The dosage form of the bactericidal composition is any one of granules, water-dispersible granules, wettable powders and suspensions.
6. Use of a bactericidal composition containing bromothionil and oxytetracycline hydrochloride according to any one of claims 1 to 5 in preventing bacterial diseases of crops.
7. The use according to claim 6, characterized in that: The crop bacterial disease is white spot disease.
8. The use according to claim 7, characterized in that: The crop is any one of corn, sorghum, rice and wheat.
9. The use according to claim 8, characterized in that: The crop is corn.
10. The use according to claim 6, characterized in that: The frequency of using the bactericidal composition is 14-20 days per time.