Suspended medicament for preventing and treating asparagus stem blight and preparation method thereof

Abstract

The invention relates to the field of fungal pesticide preparations, in particular to a suspension medicament for preventing and treating asparagus stem blight and a preparation method thereof. The suspension agent provided by the invention can effectively prevent and treat asparagus stem blight, and the prevention effect can reach 94.95%-98.99%. The microorganism combination in the suspension agent can prevent pathogenic bacteria from further invading plants, and meanwhile, the disease resistance of the plants is improved; mancozeb can effectively kill fungi and reduce the number of pathogenic bacteria; the mixed extract of myristica fragrans and acanthopanax senticosus can provide nutrition for microorganisms, and can inhibit the growth of pathogenic bacteria. According to the invention, microorganisms, chemical drugs and a mixed extract of myristica fragrans and acanthopanax are combined for the first time, the effect of the microorganisms, the chemical drugs and the myristica fragrans and acanthopanax mixed extract is found to be optimal under a specific proportion, and embodiments prove that the three components have a synergistic effect. Therefore, the invention provides a novel prevention and treatment method, the dosage of chemical agents can be reduced, and environmental pollution is reduced.

Description

Technical Field

[0001] The invention relates to the field of fungal pesticide preparations, in particular to a suspension agent for preventing and treating asparagus stem blight and a preparation method thereof. Background Art

[0002] Asparagus officinalis Linn., also known as asparagus, belongs to the Liliaceae family and the Asparagus genus. The root of asparagus is 2-3mm thick; the stem is smooth, the upper part often droops in the later stage, and the branches are relatively weak; the leaf-like branches of asparagus are clustered in 3-6 pieces, which are nearly flat cylindrical, slightly blunt, slender, often slightly curved, 5-30mm long, and 0.3-0.5mm thick; the base of the scale-like leaves has short thorns or almost no spurs; the berry is 7-8mm in diameter, red when ripe, and has 2-3 seeds; the flowering period is May-June, and the fruiting period is September-October; the young seedlings can be eaten as vegetables. Asparagus has high nutritional value, can moisten the lungs, relieve cough, and eliminate phlegm, and has the function of inhibiting tumor growth, etc., and is known as the "king of vegetables" in the international market. In recent years, with the expansion of asparagus cultivation area, the occurrence of asparagus stem blight has become more serious year by year, seriously affecting the yield and quality of asparagus.

[0003] Asparagus stem blight is also known as the "cancer" of asparagus. Its symptoms are mainly concentrated on the stems, side branches or leaves. It has the characteristics of rapid onset, easy transmission and serious losses. Once asparagus is infected, it is easy to spread over a large area, causing the entire asparagus plant to suddenly turn yellow and wither, and asparagus products cannot be effectively produced. The main pathogenic bacteria of asparagus stem blight is Phomopsis asparagi. At present, chemical pesticides such as carbendazim and thiophanate-methyl are mainly used for the prevention and control of asparagus stem blight. However, long-term use of such chemical pesticides can easily lead to drug resistance in the pathogens, and it is impossible to effectively prevent and control asparagus stem blight in a sustainable manner. Therefore, it is necessary to develop a pesticide formulation with better prevention and control effects. Summary of the invention

[0004] The purpose of the present invention is to provide a suspension agent for preventing and treating asparagus stem blight and a preparation method thereof, so as to solve the problems existing in the above-mentioned prior art. The suspension agent for preventing and treating asparagus stem blight provided by the present invention can effectively prevent and treat asparagus stem blight.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides a suspension agent for preventing and treating asparagus stem blight, comprising an active ingredient 1, an active ingredient 2, an active ingredient 3 and auxiliary materials;

[0007] The active ingredient 1 is the inactivated fermentation broth of Streptomyces albidoflavus and the inactivated fermentation broth of Bacillus pumilus; the active ingredient 2 is mancozeb; the active ingredient 3 is the mixed extract of nutmeg and acanthopanax senticosus; the auxiliary materials are wetting agent, dispersant, thickener, antifreezing agent and organic solvent.

[0008] Preferably, the mass ratio of the active ingredient 1, the active ingredient 2, the active ingredient 3 and the auxiliary materials is 1:(3 - 5):(4 - 7):1;

[0009] The mass ratio of the inactivated fermentation broth of Streptomyces albidoflavus and the inactivated fermentation broth of Bacillus pumilus in the active ingredient 1 is 1:1;

[0010] The mass ratio of the wetting agent, the dispersant, the thickener, the antifreezing agent and the organic solvent is 3:2:1:1:2.

[0011] Preferably, the preparation method of the mixed extract of nutmeg and acanthopanax senticosus is: mixing nutmeg and acanthopanax senticosus according to the mass ratio of 1:1, and then carrying out ethanol extraction to obtain the mixed extract of nutmeg and acanthopanax senticosus.

[0012] Preferably, the wetting agent is alkylphenol polyoxyethylene ether; the dispersant is sodium tripolyphosphate; the thickener is polypropylene glycol; the antifreezing agent is ethylene glycol; the organic solvent is dipropylene glycol methyl ether.

[0013] The present invention provides a preparation method of the above suspension agent, comprising the following steps:

[0014] Mixing the active ingredient 1, the active ingredient 2, the active ingredient 3 and the auxiliary materials to obtain the suspension agent.

[0015] The present invention provides an application of the above suspension agent in preventing and treating asparagus stem blight.

[0016] The present invention provides a method for preventing and treating asparagus stem blight, comprising the following steps:

[0017] Diluting the above suspension agent and spraying it on the surface of asparagus plants.

[0018] Preferably, the dilution multiple is 500 times.

[0019] Preferably, the dosage of the suspension diluent obtained after dilution is 30 L / mu.

[0020] The present invention discloses the following technical effects:

[0021] The suspension agent provided by the present invention can effectively control asparagus stem blight, and its control effect can reach 94.95%-98.99%. In the suspension agent provided by the present invention, the microbial combination can prevent the further invasion of pathogenic bacteria into the plant body and improve the disease resistance of the plant at the same time; mancozeb can effectively kill fungi and reduce the number of pathogenic bacteria; the mixed extract of nutmeg and acanthopanax can provide nutrition for microorganisms and inhibit the growth of pathogenic bacteria at the same time. The present invention combines microorganisms, chemical drugs and the mixed extract of nutmeg and acanthopanax for the first time, and finds that the three have the best effect in a specific proportion, and it is proved by examples that there is a synergistic effect among the three. Thus, the present invention provides a new control method, which can reduce the dosage of chemical agents and environmental pollution. Detailed Embodiments

[0022] The various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and implementation embodiments of the present invention.

[0023] It should be understood that the terms used in the present invention are only for describing specific embodiments and are not used to limit the present invention. In addition, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0024] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0025] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the present invention specification, which are obvious to those skilled in the art. Other embodiments obtained from the specification of the present invention are obvious to those skilled in the art. The specification and examples of the present invention are only exemplary.

[0026] Regarding the use of "comprising", "including", "having", "containing", etc. in this article, they are all open-ended terms, that is, they are intended to include but not limited to.

[0027] Without special requirements, the strains and components used in the present invention are all routinely purchased by those skilled in the art; without special requirements, the methods used in the present invention are all well-known methods to those skilled in the art.

[0028] Preparation Example

[0029] Streptomyces albidoflavus SHBCC D60726 was purchased from Wuhan Warner Biotechnology Co., Ltd., with the product number WN-BZ42238;

[0030] Preparation of inactivated fermentation broth of Streptomyces albidoflavus SHBCC D60726: After inoculating the activated Streptomyces albidoflavus SHBCC D60726 into 2×YT liquid medium (16 g of peptone, 10 g of yeast extract, 5 g of NaCl, made up to 1000 mL with tap water, natural pH), fermentation culture was carried out to obtain a fermentation broth, and the effective viable count in this fermentation broth was 1×10 7 CFU / mL; among them, the inoculation amount was 5%, the fermentation culture time was 72 h, and the temperature was 30°C; then the fermentation broth was inactivated (autoclaved at 115°C for 15 min) to obtain the inactivated fermentation broth of Streptomyces albidoflavus SHBCC D60726, abbreviated as the inactivated fermentation broth of Streptomyces albidoflavus.

[0031] Bacillus pumilus was purchased from Cangzhou Kangwei Biotechnology Co., Ltd.;

[0032] Preparation of inactivated fermentation broth of Bacillus pumilus: After inoculating the activated Bacillus pumilus into PD medium, fermentation culture was carried out to obtain a fermentation broth, and the effective viable count in this fermentation broth was 2.1×10 7 CFU / mL; among them, the inoculation amount was 10%, the fermentation culture time was 72 h, the temperature was 35°C, and the rotation speed was 220 rpm; then the fermentation broth was inactivated (autoclaved at 115°C for 15 min) to obtain the inactivated fermentation broth of Bacillus pumilus.

[0033] Bacillus velezensis was purchased from Huizao Biology, with the product number HZB136119;

[0034] Preparation of inactivated fermentation broth of Bacillus velezensis: After inoculating the activated Bacillus velezensis into broth medium, fermentation culture was carried out to obtain a fermentation broth, and the effective viable count in this fermentation broth was 2.3×10 7 CFU / mL; among them, the inoculation amount was 10%, the fermentation culture time was 72 h, the temperature was 35°C, and the rotation speed was 220 rpm; then the fermentation broth was inactivated (autoclaved at 115°C for 15 min) to obtain the inactivated fermentation broth of Bacillus velezensis.

[0035] Bacillus amyloliquefaciens was purchased from Liaoning Huaxing Biotechnology Co., Ltd.;

[0036] Preparation of inactivated fermentation broth of Bacillus amyloliquefaciens: The activated Bacillus amyloliquefaciens was inoculated into LB medium for fermentation culture to obtain a fermentation broth with an effective viable count of 1.8×10 7 CFU / mL; among them, the inoculation amount was 10%, the fermentation culture time was 72 h, the temperature was 35 °C, and the rotation speed was 220 rpm; then the fermentation broth was inactivated (inactivated by high-pressure steam at 115 °C for 15 min) to obtain the inactivated fermentation broth of Bacillus amyloliquefaciens.

[0037] Preparation of the mixed extract of nutmeg and Eleutherococcus senticosus: After pulverizing nutmeg and Eleutherococcus senticosus, they were mixed according to a mass ratio of 1:1 to obtain a mixed powder; the obtained mixed powder was mixed with 75% (volume percentage) ethanol according to a mass-to-volume ratio of 1 g:12 mL, extracted at 45 °C for 2 h, filtered, and ethanol was recovered. The obtained filtrate was the mixed extract of nutmeg and Eleutherococcus senticosus.

[0038] The above inactivated fermentation broth and the mixed extract of nutmeg and Eleutherococcus senticosus were used in the following examples.

[0039] Example 1

[0040] The components of the suspension for controlling asparagus stem blight are as follows:

[0041] Active ingredient 1 is the inactivated fermentation broth of Streptomyces albidoflavus and the inactivated fermentation broth of Bacillus pumilus, and the mass ratio of the two is 1:1;

[0042] Active ingredient 2 is mancozeb;

[0043] Active ingredient 3 is the mixed extract of nutmeg and Eleutherococcus senticosus;

[0044] The auxiliary materials are alkylphenol polyoxyethylene ether (wetting agent), sodium tripolyphosphate (dispersant), polypropylene glycol (thickening agent), ethylene glycol (anti-freezing agent) and dipropylene glycol methyl ether (organic solvent); the mass ratio of alkylphenol polyoxyethylene ether, sodium tripolyphosphate, polypropylene glycol, ethylene glycol and dipropylene glycol methyl ether is 3:2:1:1:2

[0045] The mass ratio of active ingredient 1, active ingredient 2, active ingredient 3 and auxiliary materials is 1:3:5:1.

[0046] The preparation method of the suspension for controlling asparagus stem blight is as follows:

[0047] After mixing active ingredient 2, active ingredient 3 and auxiliary materials, active ingredient 1 was added to obtain the suspension for controlling asparagus stem blight.

[0048] Example 2

[0049] Same as Example 1, except that: the mass ratio of Active ingredient 1, Active ingredient 2, Active ingredient 3 and excipient is 1:5:4:1.

[0050] Example 3

[0051] Same as Example 1, except that: the mass ratio of Active ingredient 1, Active ingredient 2, Active ingredient 3 and excipient is 1:4:6:1.

[0052] Example 4

[0053] Same as Example 1, except that: the mass ratio of Active ingredient 1, Active ingredient 2, Active ingredient 3 and excipient is 1:4:7:1.

[0054] Comparative Example 1

[0055] Same as Example 1, except that: Active ingredient 1 is the inactivated fermentation broth of Streptomyces albidoflavus and the inactivated fermentation broth of Bacillus velezensis, and the mass ratio of the two is 1:1.

[0056] Comparative Example 2

[0057] Same as Example 1, except that: Active ingredient 1 is the inactivated fermentation broth of Streptomyces albidoflavus and the inactivated fermentation broth of Bacillus amyloliquefaciens, and the mass ratio of the two is 1:1.

[0058] Comparative Example 3

[0059] Same as Example 1, except that: the mass ratio of Active ingredient 1, Active ingredient 2, Active ingredient 3 and excipient is 2:3:5:1.

[0060] Comparative Example 4

[0061] Same as Example 1, except that: the mass ratio of Active ingredient 1, Active ingredient 2, Active ingredient 3 and excipient is 1:1:1:1.

[0062] Comparative Example 5

[0063] Same as Example 1, except that: Active ingredient 2 is omitted.

[0064] Comparative Example 6

[0065] Same as Example 1, except that: Active ingredient 1 is omitted.

[0066] Comparative Example 7

[0067] Same as Example 1, except that: Active ingredient 3 is omitted.

[0068] Comparative Example 8

[0069] Same as Example 1, except that: Active ingredient 2 and Active ingredient 3 are omitted.

[0070] Comparative Example 9

[0071] Same as Example 1, except that active ingredient 1 and active ingredient 3 are omitted.

[0072] Comparative Example 10

[0073] Same as Example 1, except that active ingredient 1 and active ingredient 2 are omitted.

[0074] Experimental Example

[0075] To verify the efficacy of the suspension agents prepared in Examples 1-4 and Comparative Examples 1-10 in controlling asparagus stem blight, a control test for asparagus stem blight was carried out. An asparagus plantation planted in 2022 with the initial onset of stem blight in 2023, variety "Jinggang 701", was selected. Areas of equal area were randomly selected, with each area being 50m 2 , and each agent was repeated 3 times. At the same time, a clear water control group was set up. The spraying conditions of each area were as follows:

[0076] The suspension agents prepared in Examples 1-4 and Comparative Examples 1-10 were diluted 500 times and sprayed on the surface of asparagus plants; the dosage of the diluted suspension was 30L / mu. Spraying was carried out once every 15 days for a total of 3 times. At the same time, the disease prevention effects before and 45 days after spraying were recorded. 50 plants were randomly selected from each test area, the number of diseased plants at each level was recorded, and the disease index and control effect were calculated. The calculation formulas for the disease index and control effect are as follows:

[0077] Disease index = ∑(number of diseased plants at each level × corresponding level value) / (total number of plants surveyed × 4) × 100; The asparagus was graded according to the "DB13T1342-2010 Grading Standard for Asparagus Stem Blight".

[0078] Control effect = (disease index of the control area - disease index of the treatment area) / disease index of the control area × 100%.

[0079] The statistical results of the number of plants at each level for different treatments are shown in Table 1, and the disease index and control effect are shown in Table 2.

[0080] Table 1 Statistical results of the number of plants at each level for different treatments (plants)

[0081]

[0082] Table 2 Statistical results of the disease index and control effect

[0083]

[0084] As can be seen from Tables 1 and 2, compared with the control, the control effect of the suspension agent provided by the present invention can reach between 94.95% and 98.99%, indicating that the suspension agent provided by the present invention can effectively control asparagus stem blight. At the same time, the strains used in the present invention were screened, and it was found that when Streptomyces albidoflavus and Bacillus pumilus were combined, the control effect could be improved; the proportion of the active ingredients was screened, and it was found that after changing the dosage of each active ingredient, the efficacy of the three active ingredients would be reduced, and the effect would be significantly reduced. At the same time, in order to prove the synergistic effect among the three, the present invention set comparative examples (Comparative Example 6-Comparative Example 10) in which one or two components were omitted, and investigated their disease index and control effect. Although the control effect was also increased compared with the control group, it was still far lower than that of Examples 1-4.

[0085] In summary, the suspension agent provided by the present invention can effectively control asparagus stem blight, and its control effect can reach 94.95%-98.99%. The present invention combines microorganisms, chemical drugs and the mixed extract of nutmeg and acanthopanax senticosus for the first time, and finds that the three have the best effect at a specific ratio, and it is proved by examples that there is a synergistic effect among the three. Thus, the present invention provides a new control method, which can reduce the dosage of chemical agents and environmental pollution.

[0086] The above-described embodiments are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A suspension agent for preventing and treating asparagus stem blight, characterized in that: It includes active ingredient 1, active ingredient 2, active ingredient 3 and excipients; The active ingredient 1 is the inactivated fermentation liquid of Streptomyces albus and the inactivated fermentation liquid of Bacillus pumilus; the active ingredient 2 is mancozeb; the active ingredient 3 is the mixed extract of nutmeg and Acanthopanax senticosus; the auxiliary materials are wetting agent, dispersant, viscosity enhancer, antifreeze agent and organic solvent.

2. The suspension medicament according to claim 1, characterized in that: The mass ratio of the active ingredient 1, the active ingredient 2, the active ingredient 3 and the auxiliary material is 1:(3-5):(4-7):1; The mass ratio of the inactivated fermentation broth of Streptomyces albicans to the inactivated fermentation broth of Bacillus pumilus in the effective ingredient 1 is 1:1; The mass ratio of the wetting agent, the dispersant, the tackifier, the antifreeze agent and the organic solvent is 3:2:1:1:

2.

3. The suspension medicament according to claim 1, characterized in that: The preparation method of the mixed extract of nutmeg and acanthopanax senticosus is as follows: nutmeg and acanthopanax senticosus are mixed in a mass ratio of 1:1, and then ethanol extraction is performed to obtain the mixed extract of nutmeg and acanthopanax senticosus.

4. The suspension medicament according to claim 1, characterized in that: The wetting agent is alkylphenol polyoxyethylene ether; the dispersant is sodium tripolyphosphate; the viscosity enhancer is polypropylene glycol; the antifreeze agent is ethylene glycol; and the organic solvent is dipropylene glycol methyl ether.

5. The method for preparing the suspension medicine according to any one of claims 1 to 4, characterized in that: The following steps are involved: The active ingredient 1, the active ingredient 2, the active ingredient 3 and the auxiliary materials are mixed to obtain the suspension medicament.

6. Use of the suspension agent according to any one of claims 1 to 4 in preventing and treating asparagus stem blight.

7. A method for preventing and treating asparagus stem blight, characterized in that: The following steps are involved: The suspension agent according to any one of claims 1 to 4 is diluted and sprayed on the surface of asparagus plants.

8. The method according to claim 7, characterized in that The dilution multiple is 500 times.

9. The method according to claim 7, characterized in that: The amount of the suspension dilution obtained after dilution is 30 L / mu.

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