Agricultural composition containing matrine and Macleaya cordata extract and application thereof

Abstract

The present invention belongs to the field of pesticide technology and discloses an agricultural composition containing matrine and a Macleaya cordata extract and its application. The mass ratio of matrine to Macleaya cordata extract in the agricultural composition provided by the present invention is 2:8 or 4:6 to 6:4. The agricultural composition has a good inhibitory effect on Pseudomonas syringae pv. cucumber angular leaf spot, the pathogen of cucumber bacterial angular leaf spot. Within the mass ratio range provided by the present invention, the agricultural composition has a synergistic effect on the inhibition of Pseudomonas syringae pv. cucumber angular leaf spot. The agricultural composition provided by the present invention has a good preventive effect on cucumber bacterial angular leaf spot, with a preventive and control effect of 79.2 to 89.5%. The present invention provides a highly efficient, low-toxic, and low-pollution biopesticide for the prevention and control of cucumber bacterial angular leaf spot.

Description

Technical Field

[0001] The invention belongs to the technical field of pesticides and relates to an agricultural composition containing matrine and a Macleaya cordata extract and application thereof. Background Art

[0002] Cucumber, an annual climbing herbaceous plant of the Cucurbitaceae family, is an important cash crop, playing a crucial role in the food processing and catering industries. Cucumbers are high in water, low in calories, and rich in nutrients such as vitamin C, vitamin K, and potassium, offering diuretic, heat-clearing, and detoxifying properties. Bacterial angular leaf spot (BALS) of cucumbers is caused by infection with Pseudomonas syringae pv. lachrymans, a bacterium that affects cucumbers and reduces quality and yield. Both seedlings and mature plants can be affected, with mature leaves being the most vulnerable, primarily affecting leaves, petioles, tendrils, and fruit, and sometimes also stems. In the early stages of infection, needle-sized, light green, water-soaked spots appear on the leaves, gradually turning yellowish-brown, light brown, brown, off-white, or white. The spots are polygonal and bounded by leaf veins. When damp, milky white bacterial pus forms on the underside of the leaves. The pathogen overwinters in the soil with diseased remains or in the form of infected seeds, which are the source of initial infection the following year. The pathogen invades the host through stomata, wounds, and water pores. Conditions of temperature between 18 and 26°C and relative humidity above 75% are conducive to the occurrence of the disease.

[0003] The existing technologies for the prevention and control of bacterial angular leaf spot of cucumber mainly include agricultural prevention and control and chemical prevention and control. Agricultural prevention and control include: breeding disease-resistant varieties, crop rotation and field management. Although agricultural prevention and control can control the occurrence and spread of diseases to a certain extent, there are still some defects and limitations, such as: crop rotation is difficult to implement due to land use restrictions or crop planting habits; the breeding and renewal cycle of disease-resistant varieties is long, and it is difficult to breed disease-resistant varieties that take into account both cucumber quality and yield; field management is difficult and costly. Chemical prevention and control is the main way to prevent and control bacterial angular leaf spot of cucumber. The agents commonly used in chemical prevention and control include: copper preparations, streptomycin sulfate, zinc thiazole, kasugamycin, etc. However, the frequent use of chemical agents can easily lead to drug resistance in pathogens, reduce the prevention and control effect, and the residues of chemical pesticides on crops may affect food safety and have an impact on human health. Therefore, it is of great significance to develop high-efficiency, low-toxicity and low-pollution biological pesticides.

[0004] Matrine, an alkaloid extracted from the traditional Chinese medicinal plant Sophora flavescens, possesses multiple benefits, including insecticide, antibacterial, and plant growth promotion. Matrine can damage microbial cell walls and membranes, increasing membrane permeability and allowing leakage of intracellular substances, ultimately leading to cell inactivation and death. Sophora flavescens extract is used in agriculture to prevent and treat diseases such as gray mold in tomatoes and grapes, citrus canker, rice stripe, pear scab, cucumber and grape downy mildew, and cucumber powdery mildew.

[0005] Macleaya cordata extract is extracted from the Macleaya cordata plant, a member of the Papaveraceae family. Its main components are sanguinarine and chelerythrine, which have both bactericidal and antibacterial properties. Sanguinarine inhibits intracellular ATP synthesis, thereby affecting bacterial energy metabolism and preventing normal bacterial growth and reproduction. Chelerythrine induces the accumulation of reactive oxygen species within bacterial spores, causing damage to cellular structure and function. Macleaya cordata extract exhibits broad antimicrobial activity, effective against a variety of pathogens, including Staphylococcus aureus, Escherichia coli, Salmonella choleraesuis, and Pseudomonas aeruginosa.

[0006] At present, there are no reports on the combination of matrine and Macleaya cordata extract. Summary of the Invention

[0007] To address the above technical problems, the present invention provides an agricultural composition containing matrine and a Macleaya cordata extract and its use. The mass ratio of matrine to Macleaya cordata extract in the agricultural composition is 2:8 or 4:6 to 6:4. The agricultural composition has a strong inhibitory effect on Pseudomonas syringae pv. cucumber, the pathogen of bacterial angular leaf spot disease of cucumbers. Within the mass ratio range provided by the present invention, the agricultural composition exhibits a synergistic effect on the inhibition of Pseudomonas syringae pv. cucumber, resulting in a strong control effect of the agricultural composition on bacterial angular leaf spot disease of cucumbers. The present invention provides a highly effective, low-toxic, and low-pollution biopesticide for the control of bacterial angular leaf spot disease of cucumbers.

[0008] To achieve the technical purpose of the present invention, on the one hand, the present invention provides an agricultural composition containing matrine and Macleaya cordata extract, wherein the active ingredients are composed of matrine and Macleaya cordata extract, and the mass ratio of matrine to Macleaya cordata extract in the agricultural composition is 2:8 or 4:6 to 6:4; preferably, the mass ratio of matrine to Macleaya cordata extract in the agricultural composition is 5:5.

[0009] On the other hand, the present invention seeks to protect the use of the above-mentioned agricultural composition containing matrine and Macleaya cordata extract in inhibiting the cucumber angular leaf spot disease pathological type of Pseudomonas syringae.

[0010] Furthermore, the present invention found through indoor toxicity assay that matrine and Macleaya cordata extracts, when used alone, had an effect on the EC of Pseudomonas syringae pv. cucumber angular leaf spot. 50The values ​​were 1.36 g / L and 2.12 g / L, respectively. Combining the two can enhance the inhibitory effect of the agricultural composition on Pseudomonas syringae pv. cucumber angular leaf spot. Matrine and Macleaya cordata extract, when combined in a mass ratio of 2:8 or 4:6-6:4, showed a strong inhibitory effect on the growth of Pseudomonas syringae pv. cucumber angular leaf spot. The two botanical pesticides exhibited a synergistic effect, with SRs greater than 1.5. The optimal synergistic effect was achieved when the mass ratio of matrine and Macleaya cordata extract was 5:5, with an SR of 3.13.

[0011] On the other hand, the present invention claims protection for the use of the above-mentioned agricultural composition containing matrine and Macleaya cordata extract in preventing and treating bacterial angular leaf spot of cucumber, the pathogen of which is Pseudomonas syringaepv. lachrymans.

[0012] Furthermore, the present invention learned through pot plant experiments that within the ratio range of matrine and Macleaya cordata extract provided by the present invention, the agricultural composition has a good effect of preventing and treating bacterial angular spot of cucumber, and the prevention and treatment effect is better than any single agent. When the compound mass ratio of matrine and Macleaya cordata extract is in the range of 2:8 or 4:6~6:4 and the active ingredient dosage is 5~20 μg / mL, it has a good effect of preventing and treating bacterial angular spot of cucumber, especially when the active ingredient dosage is 20 μg / mL, the prevention and treatment effect is better, and the prevention and treatment effect of bacterial angular spot of cucumber is 79.2~89.5%; when the compound mass ratio of matrine and Macleaya cordata extract is 5:5 and the active ingredient dosage is 20 μg / mL, the agricultural composition has the best prevention and treatment effect on bacterial angular spot of cucumber, which is 89.5%. It can be seen that within the ratio range of matrine and Macleaya cordata extract provided by the present invention, the agricultural composition has a good effect of preventing and treating bacterial angular spot of cucumber.

[0013] In addition, the present invention claims protection for a method for preventing and controlling bacterial angular spot of cucumber, using the above-mentioned agricultural composition containing matrine and Macleaya cordata extract, the amount of the active ingredients of matrine and Macleaya cordata extract in the agricultural composition is 5 to 20 μg / mL; preferably, the amount of the active ingredients of matrine and Macleaya cordata extract in the agricultural composition is 20 μg / mL.

[0014] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages:

[0015] In the agricultural composition containing matrine and Macleaya cordata extract provided by the present invention, the mass ratio of matrine to Macleaya cordata extract is 2:8 or 4:6-6:4, and the composition has a better inhibitory effect on the cucumber angular leaf spot disease pathological variety of Pseudomonas syringae and has a synergistic enhancement effect, with SR greater than 1.5. In particular, when the composite mass ratio of matrine and Macleaya cordata extract is 5:5, the synergistic enhancement effect is optimal, with SR of 3.13.

[0016] The agricultural composition containing matrine and Macleaya cordata extract provided by the present invention can prevent and treat bacterial angular spot of cucumber caused by Pseudomonas syringae pv. cucumber angular spot disease. The present invention learned through pot plant experiments that within the ratio range of matrine and Macleaya cordata extract provided by the present invention, the agricultural composition has a good effect of preventing and treating bacterial angular spot of cucumber, and the prevention and treatment effect is better than any single agent. When the combined mass ratio of matrine and Macleaya cordata extract is in the range of 2:8 or 4:6 to 6:4 and the dosage of the active ingredient is 5 to 20 μg / mL, it has a good effect of preventing and treating bacterial angular spot of cucumber, especially when the dosage of the active ingredient is 20 μg / mL, the prevention and treatment effect is even better, and the prevention and treatment effect on bacterial angular spot of cucumber is 79.2 to 89.5%; when the combined mass ratio of matrine and Macleaya cordata extract is 5:5 and the dosage of the active ingredient is 20 μg / mL, the agricultural composition has the best prevention and treatment effect on bacterial angular spot of cucumber, which is 89.5%. It can be seen that within the ratio range of matrine and Macleaya cordata extract provided by the present invention, the agricultural composition has a better effect of preventing and treating bacterial angular leaf spot of cucumber.

[0017] The two active ingredients in the composition of the present invention are both botanical pesticides, which are derived from nature, do not pollute the environment and agricultural products, are relatively safe for livestock, and are not prone to drug resistance. They have good application prospects in preventing and treating cucumber bacterial angular leaf spot caused by the cucumber angular leaf spot pathogenic variant of Pseudomonas syringae. DETAILED DESCRIPTION

[0018] The technical solutions of the present invention are described below with reference to the following examples. However, the present invention is not limited to the following examples. The experimental methods and detection methods described in each example are conventional methods unless otherwise specified; the reagents and materials described are commercially available unless otherwise specified.

[0019] Matrine was purchased from Inner Mongolia Zhongnong Biochemical Technology Co., Ltd. with a specification of 10% and registration number of PD20110637.

[0020] Macleaya cordata extract was purchased from Shaanxi Xiannong Biotechnology Co., Ltd., specification: 40%, registration number: PD20211352.

[0021] NA solid culture medium: 5.0 g / L peptone, 3.0 g / L beef extract powder, 1.0 g / L yeast extract powder, 10.0 g / L sucrose, and 20.0 g / L agar powder. Mix well and adjust the pH to about 7.0. Dispense into Erlenmeyer flasks, seal, and sterilize with high-pressure steam.

[0022] NA liquid medium: Remove agar powder from NA solid medium.

[0023] Pseudomonas syringuepv. lachrymans was collected from cucumber leaves infected with bacterial angular leaf spot in a vegetable greenhouse in Linwei District, Weinan City, Shaanxi Province. It was isolated by in vitro diseased tissue separation, back-grafted for identification, purified, and inoculated onto NA solid culture medium and stored in a refrigerator at 4°C.

[0024] Example 1

[0025] This example provides an indoor toxicity test of the agricultural composition against bacterial angular leaf spot pathogens.

[0026] Matrine was dissolved in distilled water, and Macleaya cordata extract was dissolved in acetone. After dissolution, each agent was diluted with 0.1% Tween-80 aqueous solution to prepare single-dose mother liquors and mixed mother liquors with different mass ratios (1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, 9:1). Five mass concentration gradients were set for each single agent and mixture according to the equal ratio method.

[0027] Take 1 mL of the above-mentioned drugs (single or mixed) and mix them evenly with 14 mL of melted NA solid medium, then pour them into a culture dish. After solidification, a drug-containing plate is formed for later use. Pick a single colony of Pseudomonas syringae pseudomonas syringae pseudomonas spp. in 30 mL of NA liquid medium, culture at 26 ± 1 ° C and 180 r / min for 24 h to obtain seed solution, and dilute the seed solution to 1 × 10 5 0.1 mL of bacterial suspension was evenly spread on the drug-coated plate and cultured in a 28°C constant temperature incubator for 24 h. 10 mL of sterile water was then washed off the colonies on the lesions and mixed. The absorbance (OD) was measured using an ultraviolet spectrophotometer. 450 ). According to the experimental design, gradient drug-containing plates were prepared, and blank controls without drugs were set up. Each treatment was repeated 3 times. The pathogenic strains of cucumber angular leaf spot were inoculated with Pseudomonas syringae and cultured. The colonies were washed and the absorbance was measured. The inhibition rate of different treatments was calculated by the following formula. The logarithm of the drug concentration was used as the horizontal axis and the inhibition rate was used as the vertical axis. The experimental data were analyzed by regression analysis to calculate the effective inhibitory concentration (EC) of each treatment. 50 ), and the expected inhibitory concentration (EC 50exp) and the synergistic coefficient (SR), the calculation formula is as follows, and the calculation results are shown in Table 1.

[0028] Inhibition rate (%) = (OD 450 - Treatment group OD 450 )×100 / blank control group OD 450 ;

[0029] EC 50exp =(a+b) / [(a / EC 50a )+(b / EC 50 b)];

[0030] SR=EC 50exp / EC 50实际 ;

[0031] In the formula, a is the percentage of matrine in the mixture, b is the percentage of Macleaya cordata extract in the mixture, EC 50a is the effective inhibitory concentration of matrine (a) when used alone, EC 50b The effective inhibitory concentration of Macleaya cordata extract (b) when used alone. SR ≥ 1.5 indicates a synergistic effect, SR ≤ 0.5 indicates an antagonistic effect, and 0.5 < SR < 1.5 indicates an additive effect.

[0032] Table 1: Inhibitory effect of the agricultural composition on Pseudomonas syringae pv. cucumber angular leaf spot

[0033] Pharmacy Ratio <![CDATA[EC 50实际 / (g / L)]]> <![CDATA[EC 50exp / (g / L)]]> SR matrine / 1.36 / / Macleaya cordata extract / 2.12 / / Matrine: Macleaya cordata extract 1:9 1.61 2.01 1.25 Matrine: Macleaya cordata extract 2:8 1.03 1.91 1.85 Matrine: Macleaya cordata extract 3:7 1.25 1.82 1.46 Matrine: Macleaya cordata extract 4:6 0.66 1.73 2.62 Matrine: Macleaya cordata extract 5:5 0.53 1.66 3.13 Matrine: Macleaya cordata extract 6:4 0.67 1.59 2.37 Matrine: Macleaya cordata extract 7:3 1.12 1.52 1.36 Matrine: Macleaya cordata extract 8:2 1.15 1.47 1.28 Matrine: Macleaya cordata extract 9:1 1.31 1.41 1.08

[0034] As shown in Table 1, matrine and Macleaya cordata extracts alone have an effect on the EC 50 The values ​​were 1.36 g / L and 2.12 g / L, respectively. Combining the two improved the inhibitory effect of the composition against Pseudomonas syringae pv. cucumber angular leaf spot. Matrine and Macleaya cordata extract, when combined in a mass ratio of 2:8 or 4:6 to 6:4, showed a strong inhibitory effect on the growth of Pseudomonas syringae pv. cucumber angular leaf spot. The two botanical pesticides exhibited a synergistic effect, with SRs greater than 1.5. The optimal synergistic effect was achieved when the mass ratio of matrine and Macleaya cordata extract was 5:5, with an SR of 3.13.

[0035] Example 2

[0036] This example provides a pot test of the agricultural composition.

[0037] Matrine was dissolved in distilled water, and the Macleaya cordata extract was dissolved in acetone. After dissolution, it was diluted with a 0.1% Tween-80 aqueous solution. Matrine and Macleaya cordata extract were compounded in a mass ratio of 2:8 or 4:6 to 6:4, respectively. The dosage of the active ingredient in each treatment group was 5 μg / mL, 15 μg / mL, and 20 μg / mL. The spray method was used for treatment. The dosage of the drug solution in each treatment group was 5 mL. The cucumber plants sprayed with 0.1% Tween-80 sterile water were used as controls. The cucumber variety for the test was Jinyan No. 6. Healthy, growing, and uniform-sized cucumber plants were selected. Each treatment was repeated 3 times, with 6 plants per replicate. 24 hours after the drug treatment, the cucumber plants were inoculated with Pseudomonas syringae pv. cucumber angular leaf spot (1×10 5 Cucumber plants were spray-inoculated with a suspension of Pseudomonas syringae pv. cucumerinoides at a concentration of 2 mL / plant. After inoculation, the cucumber plants were placed in a moisturizing cabinet maintained at 90% relative humidity and 26°C. Ten days after inoculation, the disease was assessed, and the disease index and control efficacy of the cucumber plants in the different treatment groups were calculated using the following formula. The statistical results are shown in Table 2. The disease grading standard is:

[0038] Level 0: no lesions;

[0039] Level 1: 0% < Cucumber bacterial angular leaf spot lesions occupy ≤ 5% of the leaf surface area;

[0040] Level 3: 5%< Cucumber bacterial angular spot lesions occupy ≤25% of the leaf surface area;

[0041] Level 5: 25% < cucumber bacterial angular spot lesions occupy ≤ 50% of the leaf surface area;

[0042] Level 7: 50% < cucumber bacterial angular spot lesions occupy ≤ 75% of the leaf surface area;

[0043] Level 9: 75% < cucumber bacterial angular leaf spot lesions occupy 75% of the leaf surface area.

[0044] Disease index = [Σ(number of diseased leaves at each level × relative level value) / (total number of leaves surveyed × 9)] × 100;

[0045] Control effect (%) = (disease index of blank control group - disease index of drug-treated group) / disease index of blank control group × 100.

[0046] Table 2: Control effect of agricultural composition on bacterial angular leaf spot of cucumber

[0047]

[0048]

[0049] As shown in Table 2, when the mass ratio of matrine and Macleaya cordata extract is in the range of 2:8 or 4:6-6:4, the control effect on cucumber bacterial angular spot caused by Pseudomonas syringae pseudomonas cucumber angular spot disease is better than any single agent. When the mass ratio of matrine and Macleaya cordata extract is in the range of 2:8 or 4:6-6:4 and the dosage of the active ingredient is 5-20 μg / mL, it has a good effect on preventing and treating cucumber bacterial angular spot disease, especially when the dosage of the active ingredient is 20 μg / mL, the control effect is even better, with a control effect of 79.2-89.5% on cucumber bacterial angular spot disease; when the mass ratio of matrine and Macleaya cordata extract is 5:5 and the dosage of the active ingredient is 20 μg / mL, the agricultural composition has the best control effect on cucumber bacterial angular spot disease, which is 89.5%. The above results show that within the ratio range of matrine and Macleaya cordata extract provided by the present invention, the agricultural composition has a good effect on preventing and treating cucumber bacterial angular spot disease.

[0050] The embodiments described above are some of the embodiments of the present invention, rather than all of them. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but rather merely represents selected embodiments of the present invention. All other embodiments obtained without creative effort and through deduction and substitution by a person of ordinary skill in the art based on the concept of the present invention are within the scope of protection of the present invention.

Claims

1. An application of an agricultural composition in preventing and treating bacterial angular leaf spot of cucumber, characterized in that: The effective ingredients of the agricultural composition are composed of matrine and Macleaya cordata extract; The mass ratio of matrine to Macleaya cordata extract in the agricultural composition is 2:8 or 4:6 to 6:4; The registration number of matrine in the agricultural composition is PD20110637, and the registration number of Macleaya cordata extract is PD20211352; The pathogen of the cucumber bacterial angular spot disease is Pseudomonas syringaepv. lachrymans.

2. The application according to claim 1, characterized in that The mass ratio of matrine to the Macleaya cordata extract in the agricultural composition is 5:

5.

3. A method for using an agricultural composition in preventing and treating bacterial angular leaf spot of cucumber, characterized in that: The effective ingredients of the agricultural composition are composed of matrine and Macleaya cordata extract; The mass ratio of matrine to Macleaya cordata extract in the agricultural composition is 2:8 or 4:6 to 6:4; The registration number of matrine in the agricultural composition is PD20110637, and the registration number of Macleaya cordata extract is PD20211352; The pathogen of the cucumber bacterial angular spot disease is Pseudomonas syringaepv. lachrymans.

4. The method according to claim 3, characterized in that The method of use is spraying, and the total effective ingredient dosage of matrine and Macleaya cordata extract is 5-20 μg / mL.

5. The method according to claim 4, characterized in that: The method of use is spray treatment, and the total effective ingredient dosage of matrine and Macleaya cordata extract in the method of use is 20 μg / mL.

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