Biocontrol preparation for resisting sesame wilt

Abstract

The invention discloses a biocontrol preparation for resisting sesame fusarium wilt, and belongs to the technical field of crop disease control, the biocontrol preparation for resisting sesame fusarium wilt comprises the following raw materials: 10-20 parts of sophora flavescens, 8-15 parts of cordate houttuynia, 5-10 parts of folium artemisiae argyi, 5-10 parts of rhizoma atractylodis, 5-10 parts of salvia miltiorrhiza, and 3-8 parts of licorice. The components in the biocontrol preparation for resisting the sesame wilt effectively inhibit pathogenic bacteria of the sesame wilt through a synergistic effect, enhance the disease resistance of plants and relieve disease damage, and the biocontrol preparation has a good application prospect.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of crop disease prevention and control, and particularly relates to a biocontrol agent for resisting sesame fusarium wilt. BACKGROUND

[0002] Sesame (Sesamum indicum L.) belongs to Pedaliaceae family and Sesamum genus, is an annual herb, is one of the oldest high-quality oil crops in the world, and enjoys the reputation of "oil queen" due to containing antioxidant substances such as sesamin, sesamol and vitamin E. As one of the crops with the highest oil content in the world, the main components of fatty acids of sesame are oleic acid (18:1) and linoleic acid (18:2), and the ratio is 1:1, which is the most ideal edible oil source. China is one of the four major sesame producing countries in the world, and the single yield and total yield are also in the forefront of the world. However, due to the limitation of species characteristics, the existing varieties still have the problem of low yield caused by poor disease resistance and stress resistance.

[0003] Studies have shown that sesame is easily infected by at least 8 fungal diseases at different stages of its growth cycle. Among them, fusarium wilt is one of the main diseases of sesame. The disease mainly occurs in Northeast China, North China, Northwest China, Huanghuai and Jianghuai regions of China, and the annual incidence rate is 5%-10%, and the disease severity can reach more than 30% in serious cases, which often leads to immature sesame seeds, shrunken grains, and seeds falling off before harvest, seriously threatening and hindering the production and development of sesame.

[0004] The existing sesame fusarium wilt prevention and control technology includes agricultural prevention and control and chemical prevention and control. The agricultural prevention and control includes selecting disease-resistant varieties, reasonable crop rotation and timely sowing, etc. As an important means of sesame fusarium wilt prevention and control, agricultural prevention and control is environmentally friendly and sustainable, can reduce the dependence on chemical pesticides, and is conducive to the sustainable development of agriculture. However, the breeding and promotion of disease-resistant varieties need time, and the variation speed of pathogenic fungi is relatively fast, which may cause the original disease-resistant varieties to lose effect; although crop rotation can effectively reduce the accumulation of pathogenic fungi in the soil, in actual production, farmers may not be able to implement effective crop rotation due to limited land resources or other economic factors; timely sowing needs to be considered comprehensively according to factors such as climate conditions and soil temperature, and there is a certain difficulty in operation. Applying relevant chemical agents according to different growth periods of sesame can improve the prevention and control effect of sesame fusarium wilt, including using thiophanate-methyl or fludioxonil for seed dressing before sowing, using acetochlor emulsifiable concentrate or isopropyl amylamine for uniform spraying on the soil surface after sowing and soil closing treatment, or using carbendazim wettable powder, thiophanate-methyl suspension concentrate, chlorothalonil suspension concentrate or benomyl wettable powder during the seedling stage of sesame, etc. However, the above-mentioned chemical agents have unavoidable environmental pollution and human health risks when used.

[0005] Therefore, there is an urgent need in the art for a biocontrol agent for resisting sesame fusarium wilt that can effectively replace chemical agents. SUMMARY

[0006] The present application aims to provide a biocontrol agent for resisting sesame fusarium wilt to solve the problems of the prior art.

[0007] One of the technical solutions provided by the present application is:

[0008] A biocontrol agent for resisting sesame fusarium wilt, comprising the following raw materials in parts by mass: Sophora flavescens 10-20 parts, Houttuynia cordata 8-15 parts, Artemisia argyi 5-10 parts, Atractylodes lancea 5-10 parts, Salvia miltiorrhiza 5-10 parts, and Glycyrrhiza 3-8 parts.

[0009] Preferably, the biocontrol agent comprises the following raw materials in parts by mass: Sophora flavescens 15-20 parts, Houttuynia cordata 10-15 parts, Artemisia argyi 8-10 parts, Atractylodes lancea 8-10 parts, Salvia miltiorrhiza 8-10 parts, and Glycyrrhiza 5-8 parts.

[0010] More preferably, the biocontrol agent comprises the following raw materials in parts by mass: Sophora flavescens 15 parts, Houttuynia cordata 12 parts, Artemisia argyi 10 parts, Atractylodes lancea 10 parts, Salvia miltiorrhiza 8 parts, and Glycyrrhiza 5 parts.

[0011] The alkaloids (such as sophoramine) in Sophora flavescens have broad-spectrum antibacterial activity and can inhibit the growth of sesame fusarium wilt pathogenic bacteria; Sophora flavescens can enhance the disease resistance of plants and enhance their immune response.

[0012] The volatile oil and flavonoids in Houttuynia cordata have antiviral and antibacterial effects and can inhibit pathogenic bacteria. Houttuynia cordata can reduce the inflammatory response of plants caused by pathogenic bacteria.

[0013] The volatile oil (such as eucalyptol) in Artemisia argyi has insect-repelling and antibacterial effects and can inhibit pathogenic bacteria. In addition, Artemisia argyi can also promote plant growth and enhance its disease resistance.

[0014] The volatile oil (such as atractylone) in Atractylodes lancea has antibacterial and antifungal effects and can inhibit sesame fusarium wilt pathogenic bacteria. Atractylodes lancea can also regulate the immune system of plants and enhance disease resistance.

[0015] The tanshinone in Salvia miltiorrhiza has strong antioxidant effects and can reduce oxidative damage caused by pathogenic bacteria. Salvia miltiorrhiza can also promote plant tissue repair and accelerate disease recovery.

[0016] The glycyrrhizic acid in Glycyrrhiza has anti-inflammatory and immune-regulating effects, can reduce inflammation in plants and enhance disease resistance, and can also enhance the efficacy of other components and improve overall disease resistance.

[0017] In addition, the anthelmintic and antibacterial effects of the folium artemisiae argyi are synergistic with the antibacterial and antifungal effects of the rhizoma atractylodis, thereby further playing a role in inhibiting pathogenic bacteria.

[0018] The second technical solution provided by the application is as follows:

[0019] A preparation method of the biocontrol agent against sesame fusarium wilt comprises the following steps: ingredients are weighed according to mass fractions, the ingredients are respectively crushed and sieved, and then mixed, an ethanol aqueous solution is added, normal-temperature soaking treatment is performed, microwave extraction treatment is performed, filtration is performed, the extraction liquid is reduced in pressure and concentrated to one third of the original volume, and an additive is added to prepare the biocontrol agent against sesame fusarium wilt.

[0020] Preferably, the mass of the ethanol aqueous solution is 10-12 times the sum of the total mass of all the ingredients.

[0021] Preferably, the normal-temperature soaking treatment is performed for 4-6 hours.

[0022] Preferably, the microwave extraction treatment is performed for 30-40 minutes, the microwave power is 800-1000 W, and the extraction temperature is 50-60 DEG C.

[0023] Preferably, the additive comprises a surfactant and a stabilizer.

[0024] The third technical solution provided by the application is as follows:

[0025] The biocontrol agent against sesame fusarium wilt is applied in sesame fusarium wilt resistance, and the biocontrol agent against sesame fusarium wilt is mixed with 100-200 times of water in mass fraction and sprayed on the surface of sesame leaves.

[0026] Compared with the prior art, the application has the following advantages and technical effects:

[0027] The biocontrol agent against sesame fusarium wilt provided by the application effectively inhibits sesame fusarium wilt pathogenic bacteria, enhances plant disease resistance, reduces disease damage, and has a good application prospect. DETAILED DESCRIPTION

[0028] The various exemplary embodiments of the application are described in detail below, which should not be considered as limiting the application, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the application.

[0029] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application. Additionally, for a range of values of, for example, concentrations, amounts, and so forth, is to be understood as specifically encompassing every value falling within the range. Additionally, particular values within a stated range, or those not expressly identified, are to be understood as not being preferred or limiting in any way. These and other modifications can be made to the application in light of the foregoing detailed description, the figures and the claims.

[0030] Unless defined otherwise, 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 belongs. Although preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application. All documents mentioned herein are incorporated by reference to disclose and describe in full the methods and / or materials which are described herein. In the case of conflict between the present specification and any document incorporated herein by reference, the present specification will control.

[0031] Many modifications and variations of the present application described in the specification are possible in light of the foregoing detailed description, the figures and the claims. Other implementations of the present application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. The specification and examples given are exemplary only. It is intended to output a patent having claims which are broader than the literal scope of the specification and examples.

[0032] As used herein, the terms "comprises", "comprising", "includes", "including", "has", "having", "contains", "containing", or variations thereof, are intended to be open-ended and do not exclude other components, steps, or features.

[0033] Sesame wilt as the most frequent and most serious fungal disease in sesame production, its pathogenic species and epidemic regularity have been defined by researchers. At present, the prevention and control of the disease in agricultural practice mainly relies on chemical agents. Despite these measures, the wilt has not been effectively contained, but has shown an increasingly serious trend. The reasons behind this situation are complex: the first factor is that the sesame wilt fungus has developed strong resistance to traditional fungicides, while the development and application of new high-efficiency fungicides are still in the early stages and have not been widely popularized; secondly, the current sesame varieties have not been bred to be immune or highly resistant to wilt, and scientists have not yet fully mastered the specific pathogenesis of the disease. The effective components in traditional Chinese medicines such as Sophora flavescens, Houttuynia cordata, Artemisia argyi, Atractylodes lancea, Salvia miltiorrhiza and licorice have certain antibacterial and antiviral effects. Based on this, the present application has prepared a biocontrol agent against sesame wilt by repeatedly adjusting the amount of each component.

[0034] The embodiment of the present application provides a kind of biocontrol preparation for resisting sesame wilt, according to mass fraction, including the following raw materials: Sophora flavescens 10-20 parts, Houttuynia cordata 8-15 parts, Artemisia leaf 5-10 parts, Atractylodes 5-10 parts, Salvia miltiorrhiza 5-10 parts and licorice 3-8 parts.Exemplary, Sophora flavescens in some preferred embodiments of the present application is 15 parts, 20 parts, 10 parts and 12 parts, Houttuynia cordata is 12 parts, 10 parts, 15 parts and 8 parts;Artemisia leaf is 10 parts, 8 parts, 5 parts and 7 parts;Atractylodes is 10 parts, 8 parts, 5 parts and 7 parts;Salvia miltiorrhiza is 8 parts, 10 parts, 5 parts and 6 parts;Licorice is 5 parts, 8 parts, 3 parts and 4 parts.

[0035] The embodiment of the present application also provides a preparation method of the above-mentioned biocontrol preparation for resisting sesame wilt, comprising the following steps: taking raw materials according to mass fraction, mixing uniformly after crushing and sieving the raw materials respectively, adding ethanol aqueous solution, soaking at room temperature, microwave extraction treatment, filtering, concentrating the extract to one third of the original volume under reduced pressure, adding adjuvant, to prepare the biocontrol preparation for resisting sesame wilt.

[0036] In some preferred embodiments of the present application, the mass of the ethanol aqueous solution is 10-12 times the sum of the total mass of all raw materials.Exemplary, the mass of the ethanol aqueous solution is 10 times the sum of the total mass of all raw materials.

[0037] In some preferred embodiments of the present application, the soaking time at room temperature is 4-6 h;The microwave extraction treatment time is 30-40 min;Microwave power is 800-1000 w;Extraction temperature is 50-60 DEG C.

[0038] Soaking at room temperature is a traditional extraction method, mainly through solvent (such as anhydrous ethanol) at room temperature for a long time to soak the raw materials, so that the active ingredients are gradually dissolved out.Their role includes: (1) mild extraction, protect heat-sensitive components, in terms of the present application, alkaloids (such as matrine) in Sophora flavescens, flavonoids in Houttuynia cordata, tanshinone in Salvia miltiorrhiza and other active ingredients are sensitive to high temperature, and soaking at room temperature can avoid the damage of high temperature to these components, and ensure their biological activity.Volatile oil components (such as eucalyptol, atractylone) in Artemisia leaf and Atractylodes can also be stably dissolved out at room temperature, avoiding the loss of volatilization caused by high temperature;(2) fully dissolve water-soluble and alcohol-soluble components, anhydrous ethanol as a solvent can extract water-soluble and alcohol-soluble components in raw materials.At present, glycyrrhizin acid (water-soluble) and glycyrrhizin flavone (alcohol-soluble) in licorice can be effectively extracted.Tanshinone (alcohol-soluble) and danshensu (water-soluble) in Salvia miltiorrhiza can also be fully dissolved out.

[0039] Microwave extraction is a modern extraction technology, which uses the principle of microwave heating to quickly and efficiently extract effective components from raw materials. Its effects include: (1) microwave extraction can complete the extraction process in a short time, significantly shortening the extraction time and improving the production efficiency. Microwave heating rapidly raises the internal temperature of the raw material, breaks the cell wall, and quickly releases the effective components into the solvent; (2) microwave has selective heating effect on polar molecules (such as water and ethanol), which can quickly dissolve the polar components (such as matrine, glycyrrhizic acid and tanshinone) in the solvent and raw materials. Microwave has strong penetration and can uniformly heat the raw materials, avoiding the problem of local overheating or insufficient extraction. Microwave extraction quickly and efficiently extracts the remaining effective components on the basis of normal temperature soaking, and further destroys the cell wall structure to ensure sufficient extraction. The combination of the two methods not only ensures the extraction efficiency, but also maximizes the protection of active ingredients in the raw materials to ensure the disease resistance effect of the biocontrol agent.

[0040] In the following examples of the present application, the normal temperature soaking treatment time is 5h; the microwave extraction treatment time is 30min; the microwave power is 1000w; and the extraction temperature is 50℃.

[0041] In some preferred embodiments of the present application, the adjuvant includes a surfactant and a stabilizer. In the embodiments of the present application, the surfactant includes Tween-80 and sorbitan monooleate, wherein the addition amounts of Tween-80 and sorbitan monooleate are 0.2% and 0.3% of the volume of the concentrated liquid, respectively; and the stabilizer is ethyl cellulose, and the addition amount of ethyl cellulose is 0.2% of the mass of the concentrated liquid.

[0042] The embodiments of the present application also provide the use of the above-mentioned biocontrol agent for resisting sesame fusarium wilt. The biocontrol agent for resisting sesame fusarium wilt is mixed with 100-200 times the amount of water by mass fraction, and sprayed on the surface of sesame leaves.

[0043] The room temperature of the present application refers to 25±2℃.

[0044] In the embodiments of the present application, "parts" are "mass parts" unless otherwise specified.

[0045] Example 1: Preparation method of a biocontrol agent for resisting sesame fusarium wilt

[0046] 1) The following raw materials are weighed according to mass fraction: Sophora flavescens 15 parts, Houttuynia cordata 12 parts, Artemisia argyi 10 parts, Atractylodes 10 parts, Salvia miltiorrhiza 8 parts and Glycyrrhiza 5 parts;

[0047] 2) The above raw materials were ground and sieved (100 mesh) and mixed, then treated with an ethanol aqueous solution (ethanol and water in a volume ratio of 1:1, 10 times the total mass of the above raw materials) at room temperature for 5 hours, and then treated with microwave extraction (30 minutes, 1000, 50°C). The extract was filtered and concentrated under reduced pressure to one third of the original volume. Tween-80 (0.2%) and sorbitan monooleate (0.3%) were added to the concentrated liquid, and ethyl cellulose (0.2%) was added to the concentrated liquid, and then mixed to obtain a biocontrol agent for sesame fusarium wilt.

[0048] Examples 2-4

[0049] The amounts of the raw materials of Examples 2-4 are shown in Table 1.

[0050] Table 1

[0051]

[0052] The specific preparation method of Examples 2-4 was the same as that of Example 1, step 2).

[0053] Comparative Examples 1-6

[0054] The preparation method was the same as that of Example 1, except that the raw material components added were different, as shown in Table 2.

[0055] Table 2

[0056]

[0057] Performance test

[0058] Sesame fusarium wilt grade classification standard: sesame fusarium wilt symptoms during the plant stage are shortening of the plant internodes, dwarfing, yellowing and wilting of the leaves from the bottom to the top, browning of the roots, sometimes only half of the root system is brown, and the disease develops from the roots to the stems, with the appearance of a "half yellow" phenomenon on the side leaves of the plant, a layer of pink mold (mycelium and conidia) on the red-brown lesions on the diseased stems, small fruits, and thin seeds that are easily blown off before harvest. The specific grade classification is as follows: 0 grade: no disease on the whole plant; 1 grade: less than 1 / 3 of the leaves on the whole plant are yellow and shriveled or wilted; 2 grade: 1 / 3-1 / 2 of the leaves on the whole plant are yellow and shriveled or wilted, and the plant is slightly dwarfed; 3 grade: 1 / 2-3 / 4 of the leaves on the whole plant are yellow and shriveled or wilted, the leaves are shed, most of the stems are brown, and the plant is significantly dwarfed; 4 grade: all the leaves on the whole plant are yellow and shriveled or wilted, the leaves are shed, the stems are brown and dead.

[0059] In this invention, a 500-fold dilution of 50% carbendazim wettable powder was used as control group 1 (D1), a 500-fold dilution of 40% chlorothalonil suspension was used as control group 2 (D2), and a 1000-fold dilution of 50% benomyl wettable powder was used as control group 3 (D3). Water was used as the blank group (CK). The biocontrol agents against sesame wilt prepared in Examples 1-4 and Comparative Examples 1-6 of this invention were diluted 100 times with water. In early August, during the mature sesame plant stage, the above-mentioned agents were sprayed onto the surface of sesame leaves in different areas. According to GB / T17980.34-2000 "Guidelines for Field Efficacy Tests of Pesticides (I)", five random sampling points were taken in each plot, with four plants investigated at each point. The incidence rate and disease occurrence were investigated, the disease severity was recorded, and the disease index and control effect were calculated. The experimental results are shown in Tables 3 and 4. The efficacy calculation formula is as follows:

[0060]

[0061] The control incidence rate is the incidence rate of the CK group.

[0062] Table 3

[0063]

[0064] Table 4

[0065] D1 D2 D3 Example 1 Example 2 Control (%) 60.8 69.7 52.1 76.15 72.84 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Comparative Example 3 Control (%) 71.87 73.8 63.57 64.4 62.87 Comparative Example 4 Comparative Example 5 Comparative Example 6 Control (%) 68.7 66.87 67.01

[0066] As can be seen from Tables 3 and 4, some commercially available agents (carbendazim wettable powder and benomyl wettable powder) have shown certain resistance, which is consistent with the problems found in existing chemical reagents. The biocontrol agent for sesame wilt prepared in this invention has a comparable inhibitory effect on sesame wilt in the mature stage compared with the commercially available carbendazim wettable powder, chlorothalonil suspension and benomyl wettable powder, and can replace the above-mentioned chemical agents for the prevention and control of sesame wilt in the mature stage.

[0067] The results of comparative examples 1-6 show that, compared with example 1, when any one of the components of Sophora flavescens, Houttuynia cordata, Artemisia argyi, Atractylodes lancea, Salvia miltiorrhiza and Glycyrrhiza uralensis is removed, the biocontrol agent for sesame wilt still has an inhibitory effect on sesame wilt, but it will decrease to varying degrees. Therefore, when controlling sesame wilt, the above-mentioned raw material components proposed in this invention have a synergistic effect.

[0068] The above are merely preferred embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A biocontrol agent against Sclerotinia sclerotiorum, characterized in that, According to mass parts, the following raw materials are included: Sophora flavescens 10-20 parts, Houttuynia cordata 8-15 parts, Artemisia argyi 5-10 parts, Atractylodes lancea 5-10 parts, Salvia miltiorrhiza 5-10 parts, and licorice 3-8 parts.

2. The biocontrol formulation against Sclerotinia sclerotiorum according to claim 1, characterized in that, According to mass parts, the following raw materials are included: Sophora flavescens 15-20 parts, Houttuynia cordata 10-15 parts, Artemisia argyi 8-10 parts, Atractylodes lancea 8-10 parts, Salvia miltiorrhiza 8-10 parts, and licorice 5-8 parts.

3. The biocontrol formulation against Sclerotinia sclerotiorum according to claim 2, characterized in that, According to mass parts, the following raw materials are included: Sophora flavescens 15 parts, Houttuynia cordata 12 parts, Artemisia argyi 10 parts, Atractylodes lancea 10 parts, Salvia miltiorrhiza 8 parts, and licorice 5 parts.

4. A process for the preparation of the biocontrol preparation against Sclerotinia sclerotiorum according to any one of claims 1 to 3, characterized in that, The method comprises the following steps: taking raw materials according to mass parts, mixing the raw materials after being respectively crushed and sieved, adding an ethanol aqueous solution, soaking at room temperature, microwave extraction treatment, filtering, adding an additive to the filtrate after being concentrated to one third of the original volume under reduced pressure, and obtaining the biocontrol agent for resisting sesame blight.

5. The method for preparing the biocontrol agent against sesame wilt according to claim 4, characterized in that, The mass of the ethanol aqueous solution is 10-12 times the sum of the total mass of all raw materials.

6. The method of claim 4, wherein the biocontrol agent is prepared by the steps of: The soaking time at room temperature is 4-6 hours.

7. The method of claim 4, wherein the biocontrol agent is prepared by the steps of: a) inoculating a medium with the fungus; b) incubating the inoculated medium; c) harvesting the fungus; d) drying the harvested fungus; and e) grinding the dried fungus to a powder. The microwave extraction treatment time is 30-40 minutes, the microwave power is 800-1000 W, and the extraction temperature is 50-60℃.

8. The method of claim 4, wherein the biocontrol agent is prepared by the steps of: The additive includes a surfactant and a stabilizer.

9. Use of a biocontrol agent against Sclerotinia sclerotiorum according to any one of claims 1 to 3, characterized in that, The biocontrol agent for resisting sesame blight is mixed with 100-200 times the amount of water according to mass parts, and is sprayed on the surface of sesame leaves.

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