Application of linalyl acetate and method for preventing and treating field rice bacterial blight
By using linalyl acetate solution to inhibit rice bacterial blight pathogens, the problems of poor efficacy and environmental pollution associated with existing chemical fungicides have been solved, achieving a highly efficient and environmentally friendly control effect against rice bacterial blight.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
- Filing Date
- 2025-12-19
- Publication Date
- 2026-04-10
AI Technical Summary
Existing chemical fungicides are not very effective in controlling rice bacterial blight and pose environmental pollution problems, so there is a need to develop new environmentally friendly biological pesticides.
Linaloyl acetate was used as the sole active ingredient at a concentration of 0.3333 mg/L - 0.5333 mg/L to control rice bacterial blight. The solution was prepared by mixing it with Tween-80 and sprayed by drone to inhibit the synthesis of bacterial protein in rice bacterial blight fungus.
Linaloyl acetate has a significant inhibitory effect on rice bacterial blight pathogens, with a field control efficacy of up to 85.98%. It is green, environmentally friendly, and safe, and its inhibition rate is significantly higher than that of existing agents.
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Figure CN121817203A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of plant protection and biological control technology, specifically the application of linalyl acetate and a method for controlling bacterial blight in rice in the field. Background Technology
[0002] Rice bacterial blight is the most important bacterial disease of rice, caused by Xanthomonas oryzae, a pathogenic species of rice. Xanthomonas oryzae pv. oryzae, Xoo This disease, caused by rice blast and sheath blight, is considered one of the three major diseases of rice. It is contagious, explosive, and epidemic; the pathogen spreads with water and wind, entering through wounds and stomata, resulting in rapid spread and strong outbreaks. Due to global warming, changes in farming practices, and the degradation of disease-resistant genes in rice varieties, the disease is showing a trend of increasing severity year by year in coastal rice-growing areas and the middle and lower reaches of the Yangtze River in my country. Typically, affected rice fields experience yield reductions of 10%-30%, and in severe cases, yield reductions can reach 40%-60%, seriously threatening rice production and food security.
[0003] In current production, chemical fungicides remain the primary method for controlling rice bacterial blight. However, with widespread occurrence in the field, existing fungicides are ineffective due to the large proliferation of pathogens and high bacterial loads within diseased plants, resulting in poor control and short-lived efficacy. Furthermore, the limited variety of fungicides for controlling bacterial diseases, coupled with the inability to rotate their use, leads to rapid bacterial resistance and even worse control. The extensive use of chemical pesticides also causes environmental pollution, damages the ecosystem, kills beneficial organisms, and ultimately threatens human health. Therefore, there is an urgent need to develop novel biological pesticides that are environmentally friendly, have low toxicity, and possess novel mechanisms of action.
[0004] Linaloyl acetate is a natural substance found in bergamot, bergamot, camphor, salvia miltiorrhiza, jasmine, and many other plant essential oils; it is also found in cocoa seeds, celery, peach, kelp, and other plants. Linaloyl acetate has a floral aroma similar to lily of the valley and lavender, and is an indispensable fragrance ingredient in the preparation of high-end perfumes, widely used in the daily chemical and food industries. Studies have reported that linaloyl acetate has sedative and anti-anxiety pharmacological effects; it also possesses potential antibacterial and anti-inflammatory functions, and can improve colds, coughs, and bronchitis, especially showing good effects against pulmonary tuberculosis infections. Summary of the Invention
[0005] To address the problems in the prior art, this invention provides the application of linalyl acetate and a method for controlling rice bacterial blight in the field. Antibacterial tests have demonstrated that linalyl acetate has a strong inhibitory effect on the rice bacterial blight pathogen and can effectively control rice bacterial blight.
[0006] To achieve the above objectives, the present invention adopts the following technical solution: One of the purposes of the present application is to protect the application of linalyl acetate in preventing and treating rice bacterial leaf blight.
[0007] In the application, the concentration of linalyl acetate is 0.3333 mg / L-1.0000 mg / L; more preferably, the concentration of linalyl acetate is 0.3333 mg / L-0.5333 mg / L (it has been verified by experiments that the inhibitory effect of linalyl acetate on rice bacterial leaf blight is effective from the concentration of 0.1667 mg / L, but the effect is not good; with the increase of the concentration of linalyl acetate, the inhibitory effect of linalyl acetate on rice bacterial leaf blight is better and better, and when the concentration reaches 0.5333 mg / L, the inhibitory effect reaches 100%).
[0008] The second purpose of the present application is to protect the application of linalyl acetate in inhibiting the synthesis of bacterial leaf blight protein.
[0009] Further, in the application, the concentration of linalyl acetate is 0.1667 mg / L-0.5333 mg / L.
[0010] In the present application, the linalyl acetate used can be directly purchased from ordinary market products or can be extracted by conventional methods in the art.
[0011] The third purpose of the present application is a method for preventing and treating rice bacterial leaf blight in the field, which uses linalyl acetate or uses linalyl acetate as the only active ingredient for preventing and treating rice bacterial leaf blight in the field.
[0012] Further, the method for preventing and treating rice bacterial leaf blight in the field comprises the following steps: mixing linalyl acetate, Tween-80 and water to configure a solution; and regularly treating rice.
[0013] Further, in the method for preventing and treating rice bacterial leaf blight in the field, the final concentration of Tween-80 in the solution is 0.1 mg / L.
[0014] Further, in the method for preventing and treating rice bacterial leaf blight in the field, the concentration of linalyl acetate in the solution is 0.5333-0.8000 mg / L.
[0015] Further, in the method for preventing and treating rice bacterial leaf blight in the field, the time for treating rice is the tillering late stage to the breaking stage of rice growth.
[0016] Further, in the method for preventing and treating rice bacterial leaf blight in the field, when preventing and treating rice bacterial leaf blight in the field, a unmanned aerial vehicle is used to spray the solution, 5L-7L of the solution is sprayed per mu, and specifically, it can be 5L, 6L, 7L, etc.
[0017] The present application adopts linalyl acetate to find that linalyl acetate has inhibitory effect on the synthesis of Xoo cell protein, indicating that linalyl acetate can interfere with the cell protein metabolism of the pathogen, inhibit the growth and reproduction of the pathogen, and thus achieve the prevention and control effect of Xoo in the field.
[0018] Compared with the prior art, the present application has the following beneficial effects: (1) Linalyl acetate is a naturally occurring aromatic compound, which is widely present in plants such as bergamot and linaloe, and the raw material source is wide and green and environmentally friendly; (2) Through the study of the inhibitory effect of linalyl acetate on Xoo cell and the field control effect, it is found that at the treatment concentration of 0.3333 mg / L and 0.5333 mg / L, the inhibition rate of linalyl acetate on Xoo is 58.22% and 100%, which is significantly higher than that of the control agent kasugamycin (27.31% and 50.78%).
[0019] (3) The inhibitory effect of linalyl acetate on Xoo cell increases with the increase of the concentration of linalyl acetate, and the EC50 value of linalyl acetate on cell is 0.1992 mg / L, which is significantly better than that of the control agent kasugamycin (EC50 value is 0.4983 mg / L). Linalyl acetate has good control effect on Xoo, and the control effect of 0.5333 mg / L linalyl acetate solution on Xoo is 81.52%, which is significantly different from the control effect (71.75%) of 0.5333 mg / L kasugamycin.
[0020] (4) Linalyl acetate has inhibitory effect on the synthesis of cell protein of Xoo, and when the concentration is 0.3333 mg / L, the inhibition rate of linalyl acetate on protein cooperation is 79.32%, which is significantly higher than that of the control agent kasugamycin (7.31%). BRIEF DESCRIPTION OF DRAWINGS
[0021] Figure 1 is the inhibition effect diagram of linalyl acetate on Xoo cell at different concentrations. DETAILED DESCRIPTION
[0022] The technical solutions of the present application will be described below in conjunction with the embodiments, obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor belong to the scope of protection of the present application.
[0023] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of this application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.
[0024] It should be noted that the terminology used herein is for the purpose of describing particular implementations only and is not intended to limit the exemplary implementations according to this application. As used herein, the singular form includes the plural form unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this description, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.
[0025] In this invention, some conventional operating equipment, devices and components have been omitted or only briefly described.
[0026] Unless otherwise specified in the examples, the conditions shall be performed according to the standard conditions or the conditions recommended by the manufacturer.
[0027] In this application, unless otherwise specified, % refers to its mass percentage content.
[0028] Example 1: Test materials 1.1 Test reagents and pathogenic strains Linaloyl acetate was purchased from Jiangxi Xinsen Natural Plant Oil Co., Ltd.
[0029] 1.2 Culture medium NA solid medium (NA), NB liquid medium (NB), and Tween-80 were all domestically produced analytical grade.
[0030] 1.3 Isolation and Identification of Pathogens (1) Isolation of pathogens In 2023, rice leaf samples with typical symptoms of rice bacterial blight and bacterial ooze were collected from the field. Then, under aseptic conditions, the bacterial ooze on the sample leaves was streaked and purified on NA medium. The colony morphology of the isolated bacterial strain on NA medium was consistent with that of the rice bacterial blight pathogen reported in the standard (GB / T28078-2011). (2) Pathogen identification The experiment further extracts and separates the genomic DNA of the pathogen, uses specific primers of the Xoo to perform PCR amplification and sequencing, compares the amplified sequence with the sequence of Xoo in the GenBank library of NCBI, and the similarity of the sequence of the isolated strain with the sequence of the Xoo strain (accession number: AB045311.1) in the GenBank library is 100%, so the bacterial strain isolated in the experiment is Xoo. Xanthomonas oryzae pv. oryzae, Xoo ).
[0031] The Xoo isolated and obtained is stored at -20°C for standby.
[0032] The isolation and identification of the Xoo are prior art.
[0033] 1.4 Test rice varieties The rice variety 'Jingang 30' susceptible to Xoo is selected.
[0034] 2, Test method and result 2.1 Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of linalyl acetate on Xoo The MIC is determined by agar dilution method. Under sterile conditions, linalyl acetate and Tween-80 are filtered by a bacterial filter to obtain sterile linalyl acetate and Tween-80. 2 g of sterile linalyl acetate is mixed with 1 mL of Tween-80 to prepare a mixed solution of 667 mg / L, and 25 μL, 50 μL, 80 μL, 100 μL, 120 μL, 150 μL, and 180 μL of the solution are added to 100 mL of NA medium to prepare six drug-containing media with final concentrations of 0.1667 mg / L, 0.3333 mg / L, 0.5333 mg / L, 0.6667 mg / L, 0.8000 mg / L, and 1.000 mg / L, and pour the NA medium into a culture dish; the final concentration of the control agent chinglengmycin is also 0.1667 mg / L, 0.3333 mg / L, 0.5333 mg / L, 0.6667 mg / L, 0.8000 mg / L, and 1.000 mg / L, and the drug-free medium containing 0.100% Tween-80 is used as a control, and 80 μL of Xoo with an OD 600 value of 1 is taken by a pipette gun and uniformly coated on the drug-containing NA medium, with 5 repeats for each concentration. Place in a 28°C constant temperature incubator for dark culture, and observe the growth of the colonies after 4 days of culture. The results are shown in Figure 1 and Table 1.
[0035] Table 1 Growth of Xoo under different concentrations of linalyl acetate
[0036] Note: - no colonies; + less than 30 single colonies; ++ 30-100 colonies; +++ 100-300 single colonies; ++++ more colonies; +++++ numerous colonies.
[0037] From Table 1, linalyl acetate has a good inhibitory effect on white leaf blight. At a concentration of 0.3333 mg / L, the experimental group and the control group can be seen with the naked eye that linalyl acetate has an inhibitory effect on white leaf blight, therefore, 0.3333 mg / L is determined as the MIC value of linalyl acetate on white leaf blight (significantly lower than the MIC value of the control agent kasugamycin 0.5333 mg / L); when the concentration of linalyl acetate reaches 1.000 mg / L, no visible colonies grow on the plate, therefore, 1.000 mg / L is determined as the MBC value of linalyl acetate on white leaf blight (significantly lower than the MBC value of the control agent kasugamycin 1.500 mg / L).
[0038] 2.2 Inhibition effect determination The turbidimetric method was used to determine the inhibition effect of linalyl acetate on rice white leaf blight. The concentration with OD 600 value of 1 was used as the inoculum of white leaf blight, and the inoculum was prepared with 1% of NB medium bacteria liquid, and the drug-containing bacteria liquid was configured with concentration gradient of 0.1667 mg / L, 0.3333 mg / L, 0.5333 mg / L, 0.6667 mg / L, 0.8000 mg / L, 1.000 mg / L. Each treatment was placed on a shaking table under the condition of 28 ℃ and 170 rpm for 3 days, and the OD 600 value of the bacterial suspension was measured by spectrophotometer, and the average value was calculated. Each concentration treatment was repeated 3 times. Inhibition rate (%) = (control OD value-treatment OD value) / control OD value x 100. The results are shown in Table 2.
[0039] Table 2 Inhibition rate of rice white leaf blight under different concentrations of lavender essential oil
[0040] As can be seen from Table 2, at a concentration of 0.3333 mg / L and 0.5333 mg / L, the inhibition rate of linalyl acetate on rice white leaf blight was 58.22% and 100% respectively, which were significantly higher than the inhibition rate of the control agent kasugamycin (17.31% and 50.78%).
[0041] Table 3 Toxicity of linalyl acetate on rice white leaf blight
[0042] As can be seen from Table 3, linalyl acetate has strong inhibitory activity on rice bacterial leaf blight. With the increase of linalyl acetate concentration, its inhibitory activity is enhanced, and the EC 50 value is 0.1992 mg / L, which is significantly better than the control agent kasugamycin (EC 50 value 0.4983 mg / L).
[0043] 2.3 Effect of linalyl acetate on protein synthesis of rice bacterial leaf blight Take 2 g of sterile linalyl acetate and 1 mL of Tween-80, mix them to prepare a mixed solution of 667 mg / L. Then take 25, 50 and 80 μL of the solution and add them to 100 mL of NB medium containing bacteria to prepare three drug-containing media with final concentrations of 0.1667, 0.3333 and 0.5333 mg / L. The final concentration of the control agent kasugamycin is also 0.1667, 0.3333 and 0.5333 mg / L. Take 80 μL of bacterial solution with OD 600 value of 1 as control, and place it in a shaker at 28°C and 170 rpm for 3 days. Collect the bacterial cells of each treatment, wash them with sterile water for 3 times, centrifuge, and then dilute them to PBS buffer. After ultrasonic crushing, centrifuge to obtain the supernatant for standby.
[0044] The content of protein in the sample is determined by double antibody sandwich method. The sample is processed according to the operation process of the protein determination kit. The processed sample is placed in an enzyme marker, and the OD value of each sample is determined at a wavelength of 595 nm. The soluble protein content of each treatment sample is calculated by standard curve. Soluble protein content inhibition rate = (control soluble protein content treatment soluble protein content) / (control soluble protein content) x 100%. The results are shown in Table 4: Table 4 Inhibition of linalyl acetate on protein synthesis of bacterial leaf blight (%)
[0045] As can be seen from Table 4, linalyl acetate has strong inhibitory effect on protein synthesis of bacterial leaf blight. At concentrations of 0.1667 mg / L and 0.5333 mg / L, the inhibition rate of linalyl acetate on the protein content of bacterial leaf blight is 57.19% and 100%, which is significantly higher than that of the control agent (3.30% and 13.23%).
[0046] 2.4 Field control effect of linalyl acetate on rice bacterial leaf blight The field control effect of linalyl acetate on rice bacterial leaf blight was tested by spray method. The test variety was the susceptible variety Jinguang 30, and the rice was planted in the rice test field where bacterial leaf blight often occurred to ensure the full occurrence of bacterial leaf blight.
[0047] The test agents and dosages were as follows: 0.5333 mg / L and 0.8000 mg / L linalyl acetate solution (containing 0.1 mg / L Tween-80), 0.5333 mg / L and 0.8000 mg / L kasugamycin agent, and water as the control treatment for each treatment, which was sprayed by a plant protection unmanned aerial vehicle. Seven different treatments were designed as follows: Seven treatments were set as follows: Treatment 1: 0.5333 mg / L and 0.8000 mg / L linalyl acetate solution was sprayed at the tillering late stage and the booting stage of rice; Treatment 2: 0.5333 mg / L and 0.8000 mg / L linalyl acetate solution was sprayed at the booting stage and the jointing stage of rice; Treatment 3: 0.5333 mg / L and 0.8000 mg / L linalyl acetate solution was sprayed at the booting stage and the filling stage of rice; Treatment 4: 0.5333 mg / L and 0.8000 mg / L kasugamycin agent was sprayed at the tillering late stage and the booting stage of rice; Treatment 5: 0.5333 mg / L and 0.8000 mg / L kasugamycin agent was sprayed at the booting stage and the jointing stage of rice; Treatment 6: 0.5333 mg / L and 0.8000 mg / L kasugamycin agent was sprayed at the booting stage and the filling stage of rice; Treatment 7: water as the control treatment; Each treatment had three replicates, and 21 days after application, the leaf disease was investigated, the disease index was calculated, and the control effect was calculated.
[0048] Classification standard and calculation method of rice bacterial leaf blight disease index: 0 grade: no disease; 1 grade: disease spot expansion within 0.5 cm; 3 grade: disease spot expansion between 0.5-3 cm; 5 grade: disease spot expansion between 3-6 cm; 7 grade: disease spot expansion between 6-10 cm; 9 grade: disease spot expansion more than 10 cm.
[0049] Disease index = [∑ (number of leaves at each level × representative value at each level) / (total number of leaves surveyed × highest value)] × 100 Relative control effect (%) = (control disease index - treatment disease index) / control disease index × 100 The field control effect test results are shown in Table 5: Table 5 Field efficacy test results of linalyl acetate on rice bacterial leaf blight
[0050] As shown in Table 5, linalyl acetate has good control effect on rice bacterial leaf blight in the field. At the tillering late stage and the booting stage of rice, spraying 0.5333 mg / L-0.8000 mg / L linalyl acetate solution has the best control effect on bacterial leaf blight, and the control effect can reach 81.52%-85.98%, which is significantly better than the control agent kasugamycin (71.75%-73.49%).
[0051] In summary, the linalyl acetate provided by the present application has a significant inhibitory effect on the growth of rice bacterial leaf blight fungus, can effectively control rice bacterial leaf blight, and the control effect in the field can reach 85.98%. Linalyl acetate is a natural product of plants, easy to obtain, green and environmentally friendly, and has high safety to humans and livestock.
[0052] The above examples are only preferred embodiments of the present application, but the embodiments of the present application are not limited by the above examples, but any modification, polishing, combination made under the design and principle of the main body of the present application, the technical problems solved are considered consistent with the present application, and are within the protection scope of the present application.
Claims
1. Use of linalyl acetate in controlling rice bacterial leaf blight.
2. Use according to claim 1, characterized in that: The concentration of linalyl acetate is 0.3333 mg / L-1.0 mg / L.
3. Use according to claim 1, characterized in that: The concentration of linalyl acetate is 0.3333 mg / L-0.5333 mg / L.
4. Use of linalyl acetate in inhibiting the synthesis of bacterial leaf blight of rice.
5. Use according to claim 4, characterized in that: The concentration of linalyl acetate is 0.1667 mg / L-0.5333 mg / L.
6. A method of controlling bacterial leaf blight of rice in the field, characterized by: The method uses linalyl acetate or uses linalyl acetate as the only active ingredient for the control of rice bacterial leaf blight in the field.
7. A method for controlling bacterial leaf blight of rice in the field according to claim 6, characterized by The method comprises the following steps: Linalyl acetate, Tween-80 and water are mixed to form a solution; and rice is treated regularly.
8. The method for controlling rice bacterial leaf blight in the field according to claim 7, characterized by, The final concentration of Tween-80 in the solution is 0.1 mg / L.
9. The method for controlling rice bacterial leaf blight in the field according to claim 7, characterized by, The concentration of linalyl acetate in the solution is 0.5333-0.8000 mg / L.
10. The method for controlling bacterial leaf blight of rice in the field according to claim 7, characterized by, The time for treating rice is the tillering late stage to the breaking stage of rice growth.