Application of scabiocidin in inhibiting pathogenic bacteria of potato scab and preventing and treating potato scab

Abstract

The invention discloses an application of scabiocidin in inhibiting pathogenic bacteria of potato scab and preventing and treating the potato scab, and belongs to the technical field of pesticides. In order to develop a novel potato scab prevention and treatment means which is free of pollution to the environment and ecological system, the invention provides the application of scabestin in inhibiting pathogenic bacteria of potato scab and preventing and treating the potato scab. Experiments show that the scabiocidin has a remarkable inhibition effect on streptomyces scabies causing potato scab, and the minimum inhibitory concentration of the scabiocidin is 8 times and 256 times lower than that of shengmycin and kasugamycin in traditional medicaments for preventing and treating the potato scab; besides, the scab-killing rhzomorph has a remarkable prevention and treatment effect on potato scab caused by streptomyces scabies, the prevention and treatment effect of the scab-killing rhzomorph is superior to that of traditional Zhongshengmycin as a medicament for preventing and treating the potato scab, and the scab-killing rhzomorph can be developed and applied as an effective component of a phytopathogen bactericide or pesticide.

Description

Technical Field

[0001] The invention belongs to the technical field of pesticides, and particularly relates to the application of scabicide in inhibiting pathogenic bacteria of potato scab and preventing and controlling potato scab. Background Art

[0002] Potato scab is caused by various Streptomyces spp. and occurs widely in major potato producing areas in the world, such as the United States, China, France, and Japan. Its incidence rate in Inner Mongolia, Shanxi and other regions of my country is as high as 60%, and in severe areas it is as high as 100%. Currently, there are very few species of Streptomyces reported as pathogens of potato scab. Potato scab is mainly caused by Streptomyces scabies (S.scabies). In addition, S.acidiscabies, S.turgidiscabie, and S.europaeiscabiei can also cause potato scab.

[0003] The use of biological pesticides to control potato scab can not only avoid the carcinogenic, teratogenic, and mutagenic problems of traditional chemical pesticides, but also has the advantages of low control cost, environmental friendliness, and no pesticide residues. Among them, actinomycetes are particularly important for the discovery of biological pesticides. The products currently circulating on the market for the prevention and control of scab mainly include fungicides such as sinomenine, thiophanate-copper, quinoline copper, and some microbial preparations, but they do not have a very significant control effect. Therefore, finding a specific drug for potato scab to effectively prevent and control potato scab will be of great significance for reducing the use of chemical pesticides, achieving sustainable agricultural development, and protecting environmental safety and natural resources.

[0004] Novobiocin, also known as novobiocin, is a coumarin compound. Novobiocin was discovered due to its extremely high antibacterial activity against Staphylococcus aureus, and later it was found to have anticancer activity, but its anti-plant pathogenic bacteria activity has not been reported. Summary of the invention

[0005] In order to develop a new method for preventing and controlling potato scab without pollution to the environment and ecosystem, the present invention provides the use of scabicide in inhibiting pathogenic bacteria of potato scab and preventing and controlling potato scab. The specific technical scheme is as follows:

[0006] The first object of the present invention is to provide a use of scabicide in inhibiting the activity of pathogens causing potato scab, characterized in that the pathogen is Streptomyces scabies, and the structural formula of scabicide is shown in Formula I:

[0007]

[0008] In one embodiment of the present invention, the inhibiting the activity of pathogenic bacteria causing potato scab refers to inhibiting the growth of hyphae of the pathogenic bacteria.

[0009] The second object of the present invention is to provide the use of scabicide in preventing and treating potato scab caused by Streptomyces scab.

[0010] In one embodiment of the present invention, the application of scabicide in the application is to fill the soil or mix the seeds with the scabicide aqueous solution.

[0011] The third object of the present invention is to provide a fungicide containing the above-mentioned scabicide for use in inhibiting the activity of pathogens causing potato scab, characterized in that the pathogen is Streptomyces scab (Streptomycescabies).

[0012] In one embodiment of the present invention, the inhibiting the activity of pathogenic bacteria causing potato scab refers to inhibiting the growth of hyphae of the pathogenic bacteria.

[0013] The fourth object of the present invention is to provide a pesticide containing the above-mentioned scabicide for use in preventing and treating potato scab caused by Streptomyces scab.

[0014] The fifth object of the present invention is to provide the use of the above-mentioned scabicide in the preparation of a fungicide for inhibiting the activity of pathogenic bacteria causing potato scab, wherein the pathogenic bacteria is Streptomyces scab.

[0015] The sixth object of the present invention is to provide the use of the above-mentioned scabicide in the preparation of a pesticide for preventing and controlling potato scab caused by Streptomyces scab.

[0016] In one embodiment of the present invention, the pesticide further contains pesticide-acceptable auxiliary ingredients.

[0017] Beneficial effects of the present invention:

[0018] The scabicide of the present invention has good inhibitory activity against Streptomyces scab causing potato scab. The plate antibacterial experiment confirms that the antibacterial effect of scabicide on Streptomyces scab is significantly higher than that of zhongshengmycin, kasugamycin, thiophanate-copper and quinoline copper, which is 2.1 times that of zhongshengmycin and 4.8 times that of kasugamycin. The 96-well plate minimum inhibitory concentration detection experiment confirms that the MIC of kasugamycin is 256 μg / mL, the MIC of zhongshengmycin is 8 μg / mL, and the MIC of scabicide is only 1 μg / mL.

[0019] In addition, the scabicide described in the present invention has a significant control effect on potato scab caused by Streptomyces scab. Indoor in vitro and field experiments have confirmed that the control effect of scabicide on potato scab is better than that of traditional agents for controlling the disease, such as streptomycin and kasugamycin, and can be developed and applied as an effective ingredient of plant pathogen fungicide or pesticide. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a comparison of the plate antibacterial activities of scabicide, sinocycline, kasugamycin, thiophanate-copper, and quinoline-copper against the pathogen of potato scab. DETAILED DESCRIPTION

[0021] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific implementation methods and the accompanying drawings of the specification. The embodiments described in the present invention are not all embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0022] The experimental methods used in the following examples are all conventional methods unless otherwise specified, and the materials, reagents and instruments used are all conventional materials, reagents and instruments in the art unless otherwise specified, and can be obtained by those skilled in the art through commercial channels.

[0023] The potato scab pathogen used in the present invention is Streptomyces scabies, which is provided by Qiqihar University.

[0024] The scabicide (biocin), sinocycline, kasugamycin, thiophanate-copper and quinoline copper used in the present invention can all be purchased through commercial channels.

[0025] Example 1: Detection of the inhibitory effect of scabicide on the pathogen of potato scab

[0026] This example uses a small filter paper method to detect the inhibitory effect of scabicide on the pathogenic bacteria of potato scab. The specific operation steps are as follows:

[0027] Take Streptomyces scab spores to make 10 6 CFU / mL spore suspension, filter the mycelium with sterile cotton, absorb 200μL of the suspension and spread it on oatmeal medium (ISP3), put it on 5 sterile filter paper after drying, take 7μL of 2mg / mL zhongshengmycin, kasugamycin, thiophanate-copper, quinoline copper, and scabicide on the filter paper, inoculate the plate and invert it in a 28℃ incubator for 5 days, measure and count the diameter of the inhibition zone, and the results are shown in Figure 1 and Table 1.

[0028] Table 1 Statistical results of the inhibitory effects of scabicide and other fungicides on the pathogens of potato scab

[0029]

[0030] The results in Table 1 show that: scabicide has a significant inhibitory effect on the growth of potato scab pathogens, and the antibacterial effect is 2.1 times that of sinocycline and 4.8 times that of kasugamycin.

[0031] Example 2: Detection of the minimum inhibitory concentration of scabicide against potato scab pathogens

[0032] This example uses a small filter paper method and a 96-well plate serial dilution method to detect the minimum inhibitory concentration of scabicide for potato scab pathogens. The specific operating steps are as follows:

[0033] Pipette 200 μL 10 5 A spore suspension of Streptomyces scabies at a concentration of CFU / mL was spread on oatmeal medium (ISP3), dried and placed on sterile filter paper, 7 μL of different concentrations of scabicide was added to the filter paper, the inoculated plate was inverted and cultured in an incubator at 28°C for 5 days, the diameter of the inhibition zone was measured and counted, each treatment was repeated three times, and the concentration gradient of scabicide was: 0.125 μg / mL, 0.25 μg / mL, 0.5 μg / mL, 1 μg / mL, 2 μg / mL, 4 μg / mL, 8 μg / mL, 16 μg / mL, 32 μg / mL, 64 μg / mL, 128 μg / mL, 256 μg / mL, 512 μg / mL and 1024 μg / mL. Add 200 μL of scabicide at a concentration of 2048 μg / mL to the first well of a 96-well plate row, add 100 μL of TSB liquid culture medium to each row starting from the second well, draw 100 μL of scabicide from the first well and add it to the second well, and so on, draw 100 μL of the last well and discard it, add 100 μL of scabicide spore suspension to each well, suspend the spores of scabicide with TSB liquid culture medium, and make the spore concentration in the well be 10 6 CFU / mL, each treatment was repeated three times, zhongshengmycin and kasugamycin were selected as controls, and the results were measured and statistically analyzed after 5 days of culture in a 28°C incubator.

[0034] The results showed that the minimum inhibitory concentrations of kasugamycin, zhongshengmycin and scabracillin were 256μg / mL, 8μg / mL and 1μg / mL, respectively.

[0035] Example 3: Potted experiment of applying scabicide to control potato scab

[0036] In this example, 43 mg / L of scabicide aqueous solution was used as the experimental agent, 700-fold dilution of 3% sinoprotinin wettable powder (WP) was used as the control agent, and clean water was used as the disease control. The specific experiment is as follows:

[0037] Preparation of inoculum of pathogenic bacteria of potato scab: Streptomyces scab was propagated on ISP3 plates and cultured at 28°C for 7 days to prepare a 10 7 CFU / mL of bacterial suspension is reserved.

[0038] Seed potato germination: Select healthy seed potatoes for germination. When the sprouts reach 1.5 cm in length, disinfect their surfaces, rinse them three times with sterile water, and cut them into tubers of uniform size for later use.

[0039] The experiment was conducted in four treatments, with five pots in each treatment. Each pot contained 15 kg of soil and planted three tubers. The experiment was repeated three times. Spores of Streptomyces scab were inoculated to a final concentration of 10 6 CFU / g, 43 mg / L of scabicide aqueous solution was poured into the soil as the experimental agent, 3% sinocycline WP 700-fold diluted solution was used as the control agent, the dosage of each pot was 500 mL, and the disease grading was investigated after 80 days and the 5-level disease grading investigation results were summarized to calculate the incidence rate, disease index and control efficacy (see Table 2).

[0040] The disease index survey adopts 5-level classification and labeling, level 0: no lesions on the tubers; level 1: the area of ​​lesions on the tubers is less than 5%; level 2: the area of ​​lesions on the tubers is 5-10%; level 3: the area of ​​lesions on the tubers is 11-25%; level 4: the area of ​​lesions on the tubers is 26-50%; level 5: the area of ​​lesions on the tubers is greater than 50%. The disease index calculation formula is as follows: disease index = ∑ (number of diseased tubers at each level × representative value of disease level) / (total number of tubers surveyed × representative value of the most severe disease level) × 100. The control effect of the drug (%) = (disease index of the control group - disease index of the treatment group) / disease index of the control group × 100.

[0041] As shown in Table 2, the protective effect of scabicide on potato scab is higher than that of scabiei.

[0042] Table 2 The results of potted plant experiment on the control of potato scab by irrigating with scabicide

[0043]

[0044] Note: Disease index and relative protective efficacy are the mean ± standard error of 3 replicates. Variance analysis was performed using Duncan's multiple comparison method. The same letters indicate no significant difference at p<0.05.

[0045] Example 4: Field experiment of using scabicide to control potato scab

[0046] The experiment described in this example was conducted in the experimental field of Northeast Agricultural University. The potato variety used in the experiment was Eugene (a variety susceptible to scab disease). The experiment used a scabicide aqueous solution (final concentration of 2.5 g / mu) as the experimental agent, zhongshengmycin (active ingredient 2.5 g / mu) as the control agent, and clean water as the disease control. The specific experimental method is as follows:

[0047] The preparation of the inoculum of potato scab pathogen was the same as in Example 3. The experiment had 4 treatments, each with 3 replicates. The experiment was distributed in random blocks with a row spacing of 10 cm, a plant spacing of 8 cm, and a plot area of ​​24 m 2 , sowing in May 2024, potatoes for 10 6 The potatoes were immersed in a suspension of scab pathogenic bacteria with a concentration of CFU / mL, and the agent (43 mg / L) was applied by seed dressing. In September, 100 potatoes were randomly selected from each plot to calculate the incidence, disease index, and control effect (see Table 3). The disease grading survey method, disease index, and control effect calculation method were the same as those in Example 3.

[0048] As shown in Table 3, the protective effect of scabicide on potato scab is higher than that of scabiei.

[0049] Table 3 Field test results of the control effect of scabicide on potato scab

[0050]

[0051] Note: Disease index and relative protective efficacy are the mean ± standard error of 3 replicates. Variance analysis was performed using Duncan's multiple comparison method. The same letters indicate no significant difference at p<0.05.

[0052] The above examples show that scabicide can significantly reduce the incidence of potato scab in in vitro and field experiments, and its prevention effect is better than that of traditional anti-disease agents such as scabiei. Therefore, scabicide can be further developed into a control agent for potato scab.

[0053] Although the present invention has been disclosed as above in the form of preferred embodiments, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. The use of scabicide in inhibiting the activity of pathogenic bacteria causing potato scab, characterized in that: The pathogen is Streptomyces scabies, and the structural formula of the scabicide is shown in Formula I:

2. The use according to claim 1, characterized in that: The inhibiting of the activity of pathogenic bacteria causing potato scab refers to inhibiting the growth of hyphae of the pathogenic bacteria.

3. Use of the scabicide described in claim 1 in preventing and treating potato scab caused by Streptomyces scab.

4. The use according to claim 4, characterized in that: The application method of scabicide in the application is to fill the soil or mix the seeds with the aqueous solution of scabicide.

5. A use of a fungicide containing the scabicide of claim 1 in inhibiting the activity of pathogenic bacteria causing potato scab, characterized in that: The pathogen is Streptomyces scabies.

6. The use according to claim 5, characterized in that: The inhibiting of the activity of pathogenic bacteria causing potato scab refers to inhibiting the growth of hyphae of the pathogenic bacteria.

7. Use of a pesticide containing the scabicide as claimed in claim 1 in preventing and controlling potato scab caused by Streptomyces scab.

8. Use of the scabicide of claim 1 in the preparation of a fungicide for inhibiting the activity of pathogenic bacteria causing potato scab, characterized in that: The pathogen is Streptomyces scabies.

9. Use of the scabicide described in claim 1 in the preparation of a pesticide for preventing and controlling potato scab caused by Streptomyces scab.

10. The use according to claim 9, characterized in that: The pesticide also contains pesticide-acceptable auxiliary ingredients.

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