Compound of plant source fungicide ar-turmerone and azoxystrobin and application thereof

Abstract

The application provides a kind of plant source fungicide ar-turmerone and azoxystrobin complex agent and its application.The complex agent includes ar-turmerone fungicide and azoxystrobin, and the mass ratio of ar-turmerone fungicide to azoxystrobin is 0.27:1-0.54:1.The application also provides the application of the above-mentioned complex agent in the prevention and treatment of cucumber powdery mildew.The results of the application prove that the prevention and treatment effect of ar-turmerone fungicide and azoxystrobin in the above-mentioned complex agent on cucumber powdery mildew shows additive effect, so that good complexing effect can be realized, and cucumber powdery mildew can be effectively prevented and treated.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pesticides, in particular to a plant source fungicide ar-turmerone and azoxystrobin compound and application thereof. BACKGROUND

[0002] The current biological pesticides and other pesticides are used to reduce the number of pesticide use and dosage to control plant diseases. Mixing biological pesticides with other pesticides with different action modes not only expands the prevention and control range of diseases, but also reduces the risk of pathogen resistance. Agricultural pathogens evolve into pesticide resistance under the influence of common pesticides, and pesticide resistance brings serious agricultural, environmental and socio-economic risks. Claire et al. observed that long-term use of azole fungicides through three pathways of point mutation in CYP51 amino acid sequence, CYP51 enzyme overexpression, and overexpression of genes encoding efflux pump proteins, leading to increased pathogen resistance. If Europe stops using azole fungicides, wheat production will decrease by about 7% (9.8 million tons) in the short term, causing economic losses of up to 2.4 billion euros. Pesticide compounding is an effective method to alleviate pesticide resistance. In addition, pesticide compounding is an important way to expand the prevention and control range. At present, most of the plant source pesticides in China are mixed with broad-spectrum and high-efficiency biological pesticides or chemical pesticides for registration and use, so as to improve the quick-acting property of biological pesticides, alleviate the resistance of chemical pesticides, and expand the prevention and control range. The current compounding varieties include matrine with methomyl / cypermethrin / berberine, flavone with baicalin / aminosterol, and rotenone with pyrethrin / sumithrin / cypermethrin.

[0003] The global use of chemical synthetic pesticides has led to a series of problems such as food safety, pesticide residue exceeding the standard, target resistance, and ecological environment. The development of biological pesticides has become an important development direction of the current domestic and foreign pesticide industry. Plant source pesticides, as a kind of biological pesticide, have low residue, environmental friendliness, and target resistance, and have broad market space and application prospect. Cucumber powdery mildew is one of the main diseases of cucumber, which is usually caused by Sphaerotheca fuliginea and Erysiphe cichoracearum, and has the characteristics of frequent infection, short incubation period, and strong infectivity, which seriously affects the yield and quality. The Plant Health and Natural Products Laboratory of the School of Agriculture and Biology of Shanghai Jiaotong University extracted and separated ar-turmerone, the main active substance for preventing and treating cucumber powdery mildew, from turmeric, and developed ar-turmerone pesticide and preparation with significant effect on preventing and treating cucumber powdery mildew. However, there is no ar-turmerone compound for preventing and treating cucumber powdery mildew. SUMMARY

[0004] In view of the defects in the prior art, the present application aims to provide a compound fungicide of plant origin ar-turmerone and azoxystrobin and application thereof. Through the compound research of azoxystrobin and ar-turmerone, the additive ratio of the two for preventing and treating cucumber powdery mildew is determined, and the prevention and treatment effect is improved.

[0005] Azoxystrobin is a methoxy acrylate fungicide with a molecular weight of 403.388, and has the advantages of wide control spectrum, high efficiency and low toxicity. Azoxystrobin, as a fungicide with a wide fungicidal spectrum, has good control effect on cucumber downy mildew, turf brown spot, tobacco powdery mildew, watermelon anthracnose, tomato late blight, melon powdery mildew, turf wilt, corn small spot, lotus leaf spot, mango anthracnose and other fungal diseases. Scholars such as Wang et al. found that azoxystrobin and difenoconazole had good control effect on tobacco brown spot; scholars such as Li Wenjing found that azoxystrobin and ethylicin had additive effect on the prevention and treatment of four pathogenic fungi, i.e. helminthosporium sativum, fusarium oxysporum, colletotrichum gloeosporioides and new candida.

[0006] Azoxystrobin has been used for a long time, and there are many compound researches at present. Azoxystrobin has been formed into a compound fungicide with many pesticides and completed registration, such as propiconazole·azoxystrobin, benzyl·azoxystrobin, thiofluquinox·azoxystrobin, azoxystrobin·chlorothalonil, etc. The compound of ar-turmerone and azoxystrobin selects a new fungicide combination, expands the control range, improves the control efficiency, and provides a theoretical basis for the compound of ar-turmerone and azoxystrobin to form a new preparation in the future.

[0007] The chemical structure of ar-turmerone is shown in Figure 1 The molecular weight of ar-turmerone is 216.319. In the present application, 13% ar-turmerone is used to compound with azoxystrobin, cnidim and other drugs. The interaction between ar-turmerone and different pesticides is studied through indoor seedling stage living body bioassay, and the possibility of ar-turmerone and other pesticides is explored.

[0008] According to one aspect of the present application, a compound fungicide of plant origin ar-turmerone and azoxystrobin is provided, which comprises ar-turmerone fungicide and azoxystrobin, and the mass ratio of the ar-turmerone fungicide to the azoxystrobin is 0.27:1-0.54:1. In this ratio range, the ar-turmerone fungicide and azoxystrobin show additive effect.

[0009] Further, the mass ratio of the ar-turmerone fungicide to the azoxystrobin is 0.36:1-0.54:1

[0010] Further, the mass ratio of the ar-turmerone fungicide to the azoxystrobin is 0.54:1. The results in the embodiments of the present application show that when the volume ratio of ar-turmerone to azoxystrobin is 1:1, i.e. the weight ratio is 0.54:1, the SR value is 1.286, and the compound effect is the best.

[0011] Further, the ar-turmerone fungicide is ar-turmerone pesticide with an effective ingredient content (mass percentage) of 13%.

[0012] Further, the azoxystrobin is diluted by DMSO. DMSO is a solvent that can dissolve small polarity and is non-toxic to living organisms, and other solvents are not recommended for in vivo bioassay.

[0013] Further, the azoxystrobin has an effective ingredient concentration of 13% (mass percentage).

[0014] Further, since the ar-turmerone fungicide and the azoxystrobin cannot be mutually soluble, the complexing agent further comprises a solubilizing agent to make the two agents mutually soluble.

[0015] Optionally, the solubilizing agent is 0.1% Tween-80.

[0016] According to another aspect of the present application, there is provided a use of the complexing agent of the above-mentioned plant-derived fungicide ar-turmerone and azoxystrobin in the prevention and treatment of cucumber powdery mildew.

[0017] Further, the use comprises spraying the complexing agent on the cucumber seedlings when two true leaves of the cucumber seedlings are fully unfolded.

[0018] Further, the use comprises uniformly spraying the complexing agent on the cucumber seedlings so that the cucumber leaves are uniformly covered with agent droplets.

[0019] In addition, it is worth noting that the results of the embodiments of the present application show that, when compounded with another biological fungicide for preventing and treating cucumber powdery mildew, i.e., osthol, ar-turmerone and osthol show an antagonistic relationship, and therefore, in agricultural production, attention should be paid to avoid the use of the two pesticides together to ensure that the prevention and treatment effect is not lost.

[0020] Compared with the prior art, the present application has at least one of the following beneficial effects:

[0021] The complexing agent provided by the present application comprises ar-turmerone fungicide and azoxystrobin, and the mass ratio of ar-turmerone fungicide to azoxystrobin is 0.27:1-0.54:1. The experimental results of the present application prove that the ar-turmerone fungicide and azoxystrobin in the complexing agent show an additive effect in the prevention and treatment of cucumber powdery mildew, thereby achieving a good complexing effect, effectively preventing and treating cucumber powdery mildew, and improving the safety and effectiveness of disease control. The complexing agent can delay the resistance of the disease to a single agent or chemical pesticide, which is conducive to reducing the use of the chemical pesticide azoxystrobin, reducing costs, and protecting the ecological environment. BRIEF DESCRIPTION OF DRAWINGS

[0022] Other features, objects, and advantages of the present application will become more apparent from the following detailed description of non-limiting embodiments, made with reference to the following drawings:

[0023] Figure 1 The chemical structural formula of ar-turmerone. DETAILED DESCRIPTION

[0024] The application will be described in detail below with specific examples. The following examples will help those skilled in the art to further understand the application, but do not limit the application in any form. It should be noted that, for those skilled in the art, without departing from the concept of the application, a number of modifications and improvements can be made. These are within the scope of the present application.

[0025] EXAMPLE

[0026] 1. Materials and instruments

[0027] 1.1 Test materials

[0028] Test agents:

[0029] 13% ar-turmerone mother liquor (i.e. the effective ingredient concentration is 130 mg / mL), provided by the Plant Health and Natural Products Laboratory of the School of Agriculture and Biology, Shanghai Jiaotong University;

[0030] Azoxystrobin (purity 96.1%), provided by On Kuai (Shanghai) Chemical Co., Ltd.;

[0031] Cucumber variety: Changfeng No. 2, a powdery mildew susceptible variety (Shanghai Baoshan District Vegetable Science and Technology Promotion Station).

[0032] 1.2 Test instruments

[0033] Intelligent artificial climate chamber (Ningbo Jiangnan Instrument Factory, RXZ-500C-4);

[0034] Pipette (Dalongxingchuang Experimental Instrument (Beijing) Co., Ltd., TopPette manual single-channel adjustable pipette, 1000-5000 μL);

[0035] Electronic balance (Mettler Toledo International Trade Shanghai Co., Ltd., ME104).

[0036] 2. Test method

[0037] 2.1 Seedling cultivation

[0038] The cucumber seedlings were placed in a culture chamber at 25°C and 70% humidity, and were sprayed with a bacterial suspension or pesticide treatment when 2 true leaves were fully expanded.

[0039] 2.2 Preparation of bacterial suspension and fungal inoculation

[0040] The cucumber leaves with powdery mildew spots are washed with deionized water in a spray bottle to prepare a powdery mildew fungus suspension; select cucumber seedlings with similar growth, place them in an inoculation box, and use a spray bottle to evenly spray the fungus suspension on the cucumber leaves, so that each leaf is evenly covered with droplets of the fungus suspension. After placing the inoculation box with the lid on for 24 h, open the lid and wait for the droplets on the leaves to dry naturally.

[0041] 2.3 Preparation of the agent

[0042] (1) Preparation of the treatment agent

[0043] Azoxystrobin is dissolved with DMSO, and the effective ingredient concentration is adjusted to 13%, i.e. 130 mg / mL. The azoxystrobin solution is compounded with 13% ar-turmerone mother liquor at a volume ratio of 1:1, 1:2, and 2:3 (i.e. weight ratio of 0.54:1, 0.27:1, and 0.36:1), and 0.1% Tween-80 is added as a solubilizer.

[0044] (2) Preparation of the control agent

[0045] Solvent control: 0.1% Tween-80 water as a solvent control.

[0046] 2.4 Application of the agent

[0047] The cucumber seedlings are evenly sprayed with each agent, so that the cucumber leaves are evenly covered with droplets of the agent. Each treatment is repeated 4 times, and the test is repeated 3 times. After the droplets of the agent are allowed to dry naturally, the cucumber plants are placed in an incubator for cultivation.

[0048] 2.5 Disease investigation

[0049] The disease index of cucumber is calculated according to the method in NY / T 1156.11-2008 Pesticide Indoor Biological Determination Test Guidelines Fungicides Part 11: Pot Method for Testing Prevention of Powdery Mildew on Cucurbits. The disease investigation is performed 7 days after the application of the agent, and the disease condition is recorded.

[0050] Disease grading standards:

[0051] 0, no spots;

[0052] 1, spots accounting for less than 5% of the leaf area;

[0053] 3, spots accounting for 5% to 15% of the leaf area;

[0054] 5, spots accounting for 15% to 35% of the leaf area;

[0055] 7, spots accounting for 35% to 55% of the leaf area;

[0056] 9, spots accounting for 55% to 75% of the leaf area;

[0057] Grade 11, disease spot area more than 75% of the leaf area

[0058] Disease index and control effect were calculated according to formula (1) and formula (2).

[0059]

[0060]

[0061] Wherein, DI is disease index; N i is the number of leaves at each level; N is the total number of leaves surveyed; i is the relative level value; DC is the control effect; DI CK is the disease index of the water control group; DI T is the disease index after treatment with the agent.

[0062] Basic data statistics were processed using Excel 2016. EC50 values and 95% confidence intervals

[0063] The biological activity of A (13% ar-turmerone mother drug) and B (azoxystrobin) mixed compound was evaluated by Wadley method. The SR value (synergistic ratio) was calculated according to formula (3) and formula (4) as follows.

[0064]

[0065]

[0066] Wherein: a, b are the proportions of A and B in the compound; EC (A)50 , EC (B)50 are the EC 50 values of A and B respectively; EC 50(th) is the EC 50 value of the compound; EC 50(ob) is the measured EC 50 value of the compound. SR represents the interaction of A and B after mixing: SR < 0.5 represents antagonistic effect, 0.5 ≤ SR ≤ 1.5 represents additive effect, and SR > 1.5 represents synergistic or synergistic effect.

[0067] 3. Test results

[0068] 13% ar-turmerone mother drug and azoxystrobin solution were mixed at a volume ratio of 1:1, 1:2 and 2:3 respectively to determine the joint control effect of the compound on cucumber powdery mildew. The results of seedling bioassay are shown in Table 1.

[0069] Table 1 EC50 values and toxicity regression equations of azoxystrobin at different proportions for controlling cucumber powdery mildew

[0070]

[0071] The results showed that the toxicity of compound agents with different ratios to cucumber powdery mildew varied. When V(aromatic curcumin):V(pyraclostrobin) = 1:1, that is, m(aromatic curcumin):m(pyraclostrobin) = 0.54:1, the SR value was the largest at 1.286, indicating the strongest additive effect.

[0072] Comparative Example

[0073] 1. Materials and Instruments

[0074] 1.1 Test Materials

[0075] Test reagents:

[0076] The 13% aromatic curcumin mother drug and the 6.5% aromatic curcumin emulsifiable concentrate were provided by the Plant Health and Natural Products Laboratory of the College of Agriculture and Biology, Shanghai Jiao Tong University.

[0077] Osthole (purity 98%) was provided by Shanghai Yiji Industrial Co., Ltd.

[0078] Cucumber variety: Changfeng No. 2, a variety susceptible to powdery mildew (Shanghai Baoshan District Vegetable Science and Technology Extension Station).

[0079] 1.2 Test Instruments

[0080] Intelligent artificial climate chamber (Ningbo Jiangnan Instrument Factory, RXZ-500C-4);

[0081] Pipettes (TopPette manual single-channel adjustable pipette, 1000~5000μL, Dalongxingchuang Experimental Instruments (Beijing) Co., Ltd.);

[0082] Electronic balance (Mettler Toledo International Trading Shanghai Co., Ltd., ME104).

[0083] 2. Test methods

[0084] 2.1 Seedling cultivation

[0085] Cucumber seedlings were cultured in an incubator at 25°C and 70% humidity. Inoculation or spraying was carried out when the two true leaves were fully expanded.

[0086] 2.2 Preparation of bacterial suspension and fungal inoculation

[0087] Rinse cucumber leaves with powdery mildew lesions using deionized water from a spray bottle to prepare a powdery mildew fungal suspension. Select cucumber seedlings of similar growth and place them in an inoculation box. Spray the fungal suspension evenly onto the cucumber leaves using a spray bottle, ensuring each leaf is evenly covered with droplets of the suspension. After covering the inoculation box and leaving it for 24 hours, open the lid and allow the droplets on the leaves to air dry naturally.

[0088] 2.3 Formulation of the agent

[0089] (1) Formulation of the treatment agent

[0090] Osthole was dissolved with DMSO, and the concentration of the active ingredient was adjusted to 13%, i.e. 130 mg / mL. The osthole solution was mixed with 13% ar-turmerone base at a volume ratio of 1:1, 1:2, and 2:3 (i.e. a mass ratio of 0.89:1, 0.44:1, and 0.59:1), respectively, and 0.1% Tween-80 was used as a cosolvent.

[0091] (2) Formulation of the control agent

[0092] Positive control: 6.5% ar-turmerone emulsion was diluted 600 times as a positive control.

[0093] Solvent control: 0.1% Tween-80 water was used as a solvent control.

[0094] 2.4 Application of the agent

[0095] Each agent was uniformly sprayed on the cucumber seedlings, so that the cucumber leaves were uniformly covered with agent droplets. Each treatment was repeated 4 times, and the test was repeated 3 times. After the agent droplets were naturally air-dried, the cucumber plants were placed in an incubator for incubation.

[0096] 2.5 Disease investigation

[0097] The disease index of the cucumber was statistically analyzed according to the NY / T 1156.11-2008 Pesticide Indoor Biological Determination Test Guidelines Fungicides Part 11: Prevention of Cucumber Powdery Mildew Test Pot Method. The disease investigation was performed 7 days after the application of the agent, and the disease condition was recorded.

[0098] Disease classification standards:

[0099] 0, no disease spot;

[0100] 1, disease spot accounting for less than 5% of the leaf area;

[0101] 3, disease spot accounting for 5% to 15% of the leaf area;

[0102] 5, disease spot accounting for 15% to 35% of the leaf area;

[0103] 7, disease spot accounting for 35% to 55% of the leaf area;

[0104] 9, disease spot accounting for 55% to 75% of the leaf area;

[0105] 11, disease spot accounting for more than 75% of the leaf area

[0106] The disease index and the control effect were calculated according to the above-mentioned formula (1) and formula (2).

[0107] The basic data statistics were processed using Excel 2016. The EC50 values and 95% confidence intervals were calculated using DPS software.

[0108] The biological activity of A (13% ar-turmerone mother liquor) mixed with B (osthole) (compound preparation) was evaluated by Wadley method, and the SR value (synergistic ratio) was calculated according to the above formula (3) and formula (4).

[0109] 3. Test results

[0110] The disease investigation results are shown in Table 2.

[0111] Table 2 EC50 values and toxicity regression equations of osthole in various proportions for preventing and treating cucumber powdery mildew

[0112]

[0113] According to the experimental results in Table 2, the EC50 (ob) values of osthole and ar-turmerone at the ratios of 1:1, 1:2 and 2:3 are 0.1483 mg / mL, 0.1893 mg / mL and 0.1858 mg / mL respectively, and the SR values at the ratios of 1:1, 1:2 and 2:3 are all SR < 0.5, which shows antagonistic effect, and ar-turmerone and osthole should not be used at the same time to ensure that the prevention and treatment effect is not lost.

[0114] The specific embodiments of the present application are described above. It should be understood that the present application is not limited to the specific embodiments described above, and various modifications or changes can be made by those skilled in the art within the scope of the claims, which does not affect the essential content of the present application. The above preferred features can be used in combination in the case of not conflicting with each other.

Claims

1. Application of a compound of a plant-derived fungicide, ar-turmerone, and azoxystrobin, in the prevention and treatment of cucumber powdery mildew, wherein the compound comprises ar-turmerone fungicide and azoxystrobin, and the mass ratio of the ar-turmerone fungicide to the azoxystrobin is 0.27:1-0.54:

1.

2. Use according to claim 1, characterized in that, The application comprises spraying treatment when two true leaves of cucumber seedlings are fully unfolded.

3. Use according to claim 1, characterized in that, The application comprises uniformly spraying the compound on cucumber seedlings, so that the cucumber leaves are uniformly covered with droplets of the fungicide.

4. Use according to claim 1, characterized in that, The mass ratio of the ar-turmerone fungicide to the azoxystrobin is 0.54:

1.

5. The use according to claim 1, characterized in that, The ar-turmerone fungicide is ar-turmerone mother medicine with an effective ingredient content of 13%.

6. Use according to claim 1, characterized in that, The azoxystrobin is diluted by DMSO.

7. The use according to claim 1, characterized in that, The effective ingredient concentration of the azoxystrobin is 13%.

8. The use according to claim 1, characterized in that, The compound further comprises a cosolvent.

9. Use according to claim 8, characterized in that, The cosolvent is 0.1% Tween-80.

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