阅读说明：本技术 一种用于防治梨炭疽病的保鲜剂及其应用 (Preservative for preventing and treating anthracnose of pears and application of preservative ) 是由 吴祥为 唐勇峰 胡奎奎 李晓萌 于 2021-10-11 设计创作，主要内容包括：本发明提供一种用于防治梨炭疽病的保鲜剂,保鲜剂中活性成分是由药剂A、B和竹醋液组成,药剂A和B组合为吡唑醚菌酯和戊唑醇组合、或者、吡唑醚菌酯和甲基硫菌灵组合。可有效控制炭疽病对梨储藏期的危害,有效减少药剂的使用量,延缓炭疽病对抗药剂的抗药性,提高保鲜效果,降低贮藏损失,具有显著的经济和社会效益。(The invention provides an antistaling agent for preventing and treating anthracnose of pear, active ingredients in the antistaling agent consist of A, B medicaments and bamboo vinegar, and the combination of the A medicaments and the B medicaments is the combination of pyraclostrobin and tebuconazole or the combination of pyraclostrobin and thiophanate-methyl. The method can effectively control the harm of the anthracnose to the pear storage period, effectively reduce the using amount of the medicament, delay the drug resistance of the anthracnose to the medicament, improve the preservation effect, reduce the storage loss and have remarkable economic and social benefits.)

1. An antistaling agent for preventing and treating pear anthracnose is characterized in that active ingredients in the antistaling agent consist of A, B medicaments and bamboo vinegar, and the combination of the medicaments A and B is the combination of pyraclostrobin and tebuconazole or the combination of pyraclostrobin and thiophanate-methyl.

2. The preservative according to claim 1, wherein the mass ratio of the agent A to the agent B is 25: 1-1: 10, the mass percentage of the chemical pesticide is 1-5%, and the mass percentage of the bamboo vinegar is 60-80%.

3. The preservative according to claim 2, wherein the preservative is in the form of an aqueous emulsion and an aqueous suspension.

4. The preservative for preventing and treating anthracnose of pears according to any one of claims 1 to 3, which is characterized in that the application method is spraying or fruit soaking.

Technical Field

The invention belongs to the field of plant preservatives, and particularly relates to a preservative for preventing and treating pear anthracnose and application thereof.

Background

Anthracnose of pear, also called bitter rot, is a disease which is caused by infection of colletotrichum trabeculosum and occurs on pear. The pathogen is colletotrichum gloeosporioides sPhenz. Pathogenic bacteria overwinter on the runt fruits or diseased branches with mycelium, and the pear trees are infected when the conditions are proper in the next year. The disease gradually occurs along with the reduction of the vitality of the pears after picking, and the disease is usually continuously enlarged during the storage period and even a large amount of fruits are rotten, so that certain economic loss is caused. Anthracnose of pear occurs generally and occurs in pear producing areas of the world.

At present, the pear storage and preservation method comprises a chemical preservative, a natural preservative, a biological preservative, modified atmosphere storage and the like. The chemical preservative has the advantages of low cost, high control effect and the like, and is still an important preservation and corrosion prevention means at present. Although the chemical preservative can play a role in preserving and refreshing, most of the chemical preservatives have toxicity and residues with different degrees and have potential harm to human bodies.

The bamboo vinegar liquid belongs to a natural preservative, is a byproduct generated in the carbonization and pyrolysis processes of bamboo, consists of various components such as organic acid, phenols, ketones, alcohols, lipids and the like, has a strong inhibiting effect on various bacteria and fungi, and is a good development material of a natural source preservative. Therefore, through compounding the bamboo vinegar liquid and the chemical preservative, the preservative for preventing and treating the anthracnose of the pear is developed and researched, and the preservative plays a positive role in reducing the using amount of the chemical preservative and enhancing the preservation effect.

Disclosure of Invention

The invention provides a preservative for preventing and treating anthracnose of pears and application thereof. The invention provides the following technical scheme:

an antistaling agent for preventing and treating anthracnose of pear, the active component in antistaling agent is composed of A, B and bamboo vinegar, the combination of A and B is pyraclostrobin and tebuconazole, or pyraclostrobin and thiophanate-methyl. The preservative comprises a traditional Chinese medicine agent A and a medicine agent B according to the mass ratio of 25: 1-1: 10, wherein the mass percentage of chemical pesticides is 1-5%, and the mass percentage of bamboo vinegar liquid is 60-80%. The preferable mass ratio of the medicament A to the medicament B is 10: 1-1: 10, 5: 1-1: 5.

The combination of the medicament A and the medicament B which is preferred in the invention is pyraclostrobin and tebuconazole.

The dosage form of the preservative is aqueous emulsion and aqueous suspension. The application method of the preservative is spraying or fruit soaking. Can effectively prevent and control the diseases caused by the anthracnose of the pear and play a role in reducing the use of chemical fresh-keeping.

The formulation of the invention is an aqueous emulsion, and the weight percentage of each component is as follows:

the processing steps of the medicament composition aqueous emulsion are as follows: mixing the raw medicine and the solvent phase, adding into a high-speed homogenizing kettle, uniformly mixing by high-speed shearing, and stirring for 0.1-0.5h to form a uniform oil phase; adjusting the pH value of the bamboo vinegar (pH is less than 7), mixing with emulsifier, and stirring with a stirrer to obtain uniform water phase; opening a high-speed shearing machine in the high-speed homogenizing kettle, slowly adding the water phase into the high-speed homogenizing kettle, and homogenizing for 0.5-2h to obtain the composition aqueous emulsion.

The formulation of the invention is an aqueous suspending agent, and the weight percentage of each component is as follows:

the processing steps of the aqueous suspending agent of the medicament composition are as follows: mixing the auxiliary agent and water, uniformly mixing by high-speed shearing, adding the raw medicines of the medicine A and the medicine B, and grinding for 0.5-2 hours in a sand mill to ensure that the particle size of particles is below 5 mu m, thereby preparing the composition water suspending agent.

The solvent is one or more of cyclohexane, dichloroethane, N-hexane, ethyl acetate, chloroform, cyclohexane, acetone, N-dimethylformamide, toluene, carbon tetrachloride and ethylene glycol.

The thickening agent is one or more of magnesium aluminum silicate, xanthan gum, hydroxymethyl cellulose, gum arabic, polyvinylpyrrolidone and xanthan gum.

The emulsifier is one or more of alkylphenol polyoxyethylene ether phosphate, alkylphenol polyoxyethylene formaldehyde condensate, diphenyl phenol polyoxyethylene polyformaldehyde condensate, diphenyl phenol polyoxyethylene polyoxypropylene ether, tween-60, NNO, MOA-7, 601, A-110, TX-10, Ningru No. 700 and 5202.

The invention has the beneficial technical effects that:

the preservative prepared by the invention can effectively control the harm of anthracnose to the pear storage period, effectively reduce the using amount of the medicament, delay the drug resistance of the anthracnose to the medicament, improve the preservation effect, reduce the storage loss and have remarkable economic and social benefits.

Drawings

FIG. 1: fresh-keeping effect diagram of compound combination 1 on pear anthracnose

FIG. 2: compound combination 3 fresh-keeping effect picture for pear anthracnose

Detailed Description

The present invention will be described in detail with reference to examples. In the present invention, the following examples are intended to better illustrate the present invention and are not intended to limit the scope of the present invention. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Example 1 toxicity of bamboo Vinegar solution and Fungicide against test plant pathogenic bacteria

Test crop germs: anthracnose of pear

The determination method comprises the following steps: the toxicity of the bamboo vinegar liquid and the bactericide to the pathogenic bacteria of the test plants is measured by adopting a hypha growth rate method.

The bamboo vinegar to be tested was diluted with sterile water to an appropriate concentration. Taking 1mL of diluted bamboo vinegar, adding into 24mL of PDA culture medium, mixing, and pouring into a culture dish to obtain a culture plate containing bamboo vinegar. PDA medium plates with the same amount of sterile water added were used as blank Control (CK).

The bactericide to be tested is dissolved in acetonitrile and diluted to a proper concentration, 1mL of bactericide solution is respectively taken and evenly mixed with 24mLPDA culture medium, and the mixture is poured into a culture dish to prepare the medicine-containing culture medium with different concentrations. PDA medium plates with the same amount of acetonitrile added were used as blanks (CK).

Under the aseptic environment, a puncher is used for preparing a test strain cultured by the solid PDA culture medium into a bacterial cake with the diameter of 5.0 mm. Inoculating the test pathogenic bacteria cake (hypha face down). And (4) inverting the inoculated PDA culture medium plate, and culturing in a thermostat at 28 ℃. Culturing until the CK hyphae grow to about 85% of the culture dish, and measuring the colony diameter by using an anti-vernier caliper. And (4) calculating the hypha growth inhibition rate, and obtaining a regression equation, a correlation coefficient and EC50 of the toxicity of each single agent.

TABLE 1 toxicity of single agent and bamboo vinegar on pear anthracnose

Example 2 testing of the Combined Effect of combinations

Various proportioning combinations are designed through orthogonal experiments according to the EC50 of each single agent and the bamboo vinegar, and the proportioning combinations are diluted to proper concentration by sterile water. Taking 1mL of diluted bamboo vinegar, adding into 24mL of PDA culture medium, mixing, and pouring into a culture dish to obtain a culture plate containing bamboo vinegar. PDA medium plates with the same amount of sterile water added were used as blank Control (CK).

Under the aseptic environment, a puncher is used for preparing a test strain cultured by the solid PDA culture medium into a bacterial cake with the diameter of 5.0 mm. Inoculating the test pathogenic bacteria cake (hypha face down). And (4) inverting the inoculated PDA culture medium plate, and culturing in a thermostat at 28 ℃. Culturing until the CK hyphae grow to about 85% of the culture dish, and measuring the colony diameter by using an anti-vernier caliper. And (4) calculating the hypha growth inhibition rate, and solving a regression equation, a correlation coefficient, an EC50 and a cotoxicity coefficient of each single agent.

The combined toxicity of the compound agent is judged by using the CTC value of the co-toxicity coefficient. The CTC value is less than 80, the antagonism is achieved, the addition effect is achieved when the CTC value is between 80 and 120, the 120-200-beta-cyclodextrin has a certain synergistic effect, and the obvious synergistic effect is achieved when the CTC value is more than 200.

TABLE 2 orthogonal experimental design sheet

Note: 1, 2, 3 and 4 in the list represent multiples of EC50, and taking the experimental group No. 1 as an example A1B1C1, the concentration ratio of each component in the experiment is 1 time of bamboo vinegar EC 50: 1 time of medicament AEC 50: 1 time of medicament B EC 50; A. b is thiophanate-methyl, tebuconazole or pyraclostrobin.

TABLE 3 virulence of the combination (pyraclostrobin tebuconazole bamboo vinegar) against pear anthracnose

As can be seen from the table 3, the combined toxicity index CTC of the compounded combination (pyraclostrobin, tebuconazole and bamboo vinegar) is mostly more than 200, which indicates that the compounded agent 1 has obvious synergistic effect.

TABLE 4 virulence of the combination (pyraclostrobin-thiophanate-methyl-bamboo vinegar) against anthracnose of pear

As can be seen from Table 4, the combined toxicity indexes CTC of the compounded combination (pyraclostrobin, thiophanate-methyl and bamboo vinegar) are all larger than 120, which indicates that the compounded agent has obvious synergistic action.

Example 3 in vivo bacteriostatic Activity assay

The test method comprises the following steps: in the embodiment, the fruit soaking treatment is carried out on the formula and different medicaments, and after the fruit is dried, each fruit is perforated (3mm in depth) and 3 holes are punched. 5mm mushroom cakes were attached to each well and 6 pears were treated per group. The culture is carried out for 6 days at room temperature, the diameter of the disease spot is measured, and the control effect of the medicament on the anthracnose is verified.

Testing the medicament:

TABLE 5 composition of compounding agent (1L) and control object

Name of compound group

Bamboo vinegar liquid

Thiophanate-methyl

Tebuconazole

Pyraclostrobin (Kresoxim-methyl)

Auxiliary agent

Dosage forms

Control object

(compounding agent 1) pyraclostrobin, tebuconazole and bamboo vinegar

745.7ml

/

11.4g

17g

225.9g

Aqueous emulsion

Anthracnose

(compounding agent 3) pyraclostrobin, thiophanate-methyl, bamboo vinegarLiquid for treating urinary tract infection

844.6ml

43.9g

/

43.9g

67.6g

Suspending agent

Anthracnose

Comparison of the medicaments: 30% pyraclostrobin emulsion in water, 43% tebuconazole missible oil and 70% thiophanate methyl wettable powder

Test results

Table 6: analysis table of in vivo experiment prevention and treatment effects of compound agent 1 and control group

As can be seen from Table 6, the in vivo control effect shows that when the preparation 1 is diluted according to 800, the control effect is slightly lower than 4000 times of 30% pyraclostrobin and is equivalent to 4000 times of 43% tebuconazole; when diluted 1000 times, the control effect is obviously superior to 5000 times of 43 percent tebuconazole.

The compound agent 1 is 800 times diluted liquid medicine, each liter contains 0.014g of pyraclostrobin and 0.021g of tebuconazole, the single dose of pyraclostrobin 1200 times diluted liquid medicine, each liter contains 0.250g of pyraclostrobin, and the total amount of active ingredients of chemical pesticides is reduced by 85.6%; the single-dose tebuconazole 4000-fold diluted liquid medicine contains 0.107g of tebuconazole per liter, and the total amount of active ingredients of the chemical pesticide is reduced by 67.0 percent;

the 1000-time diluted liquid medicine of the compound agent 1 contains 0.011g of pyraclostrobin and 0.017g of tebuconazole per liter, and compared with the 2400-time diluted liquid medicine of single pyraclostrobin, the 1000-time diluted liquid medicine of the compound agent contains 0.125g of pyraclostrobin per liter, and the total amount of active ingredients of chemical pesticides is reduced by 77.3 percent; compared with 5000 times diluted medicinal liquid of single dose of tebuconazole, each liter of tebuconazole contains 0.086g, and the total amount of active ingredients of the chemical pesticide is reduced by 67.0 percent; the compound agent 1 has good control effect on pear anthracnose and simultaneously achieves the purpose of pesticide reduction.

Table 7: analysis table of in vivo experiment prevention and treatment effects of compound agent 3 and control group

As can be seen from Table 7, when the compound 3 is diluted by 800 times, the control effect is slightly lower than that of 800 times of 80% thiophanate-methyl solution and is equivalent to 1200 times of 30% pyraclostrobin; when diluted 1200 times, the control effect is obviously 2400 times better than that of 30 percent pyraclostrobin.

The 800-time diluted liquid medicine of the compound agent 3 contains 0.055g of thiophanate methyl and 0.055g of pyraclostrobin per liter, and compared with the 800-time diluted liquid medicine of single-dose thiophanate methyl, the per liter contains 0.875g of thiophanate methyl, the total amount of the effective components of the chemical pesticide is reduced by 87.5 percent; compared with a single dose of pyraclostrobin 1200 times diluted liquid medicine, each liter contains 0.250g of pyraclostrobin, and the total amount of active ingredients of the chemical pesticide is reduced by 56.1%;

1200 times of the diluted liquid medicine of the compound agent 3, each liter contains 0.044g of thiophanate methyl and 0.010g of tebuconazole, and each liter contains 0.583g of thiophanate methyl compared with the 1200 times of the diluted liquid medicine of the single-dose thiophanate methyl, and the total amount of the active ingredients of the chemical pesticide is reduced by 87.5 percent; compared with 2400 times of single dose of pyraclostrobin diluted liquid medicine, each liter contains 0.125g pyraclostrobin, and the total amount of active ingredients of the chemical pesticide is reduced by 41.5%;

the compound agent 3 is shown to have better control effect on pear anthracnose and achieve the purpose of pesticide reduction.

Example 4 Combined action test of bamboo vinegar on bactericidal combinations

Table 8: the toxicity of the pear anthracnose is compared by adding the bamboo vinegar liquid in the compound 1 and not adding the bamboo vinegar liquid

Experimental group	Regression equation	Correlation coefficient	Co-toxicity coefficient
				Adding bamboo vinegar	Y＝1.6745x-0.7154	0.9991	251.58
Bamboo vinegar liquid is not added	Y＝1.3985X+0.0403	0.97944	185.11

Bamboo vinegar EC50, pyraclostrobin EC50, tebuconazole EC50 of 4: 1: 2

According to the invention, the pesticide prepared by taking the bamboo vinegar as the main body is firstly prepared, and as can be seen from the table 8, the co-toxicity coefficient of the pesticide is obviously improved by using the bamboo vinegar in the compound 1.

Table 9: the toxicity of the pear anthracnose is compared by adding the bamboo vinegar liquid in the compound 3 and not adding the bamboo vinegar liquid

Experimental group	Regression equation	Correlation coefficient	Co-toxicity coefficient
				Adding bamboo vinegar	Y＝2.8501X-4.387	0.9547	289.96
Bamboo vinegar liquid is not added	Y＝4.4942X-10.437	0.965	209.43

Bamboo vinegar EC50, pyraclostrobin EC50, thiophanate-methyl EC 50-2: 1

According to the invention, the pesticide prepared by taking the bamboo vinegar as the main body is firstly prepared, and as can be seen from the table 9, the co-toxicity coefficient of the pesticide is obviously improved by using the bamboo vinegar in the compound 3.