阅读说明：本技术 一种用于防治梨炭疽病和梨轮纹病的保鲜剂及其应用 (Preservative for preventing and treating pear anthracnose and pear ring rot and application thereof ) 是由 吴祥为 张召贤 李晓萌 郭玉瑞 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和梨轮纹病的保鲜剂,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组合为甲基硫菌灵和戊唑醇组合。可有效控制炭疽病、轮纹病对梨储藏期的危害,有效减少药剂的使用量,延缓炭疽病、轮纹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 a preservative for preventing and treating anthracnose and pear ring rot, active ingredients in the preservative consist of A, B medicaments and bamboo vinegar, and the combination of A medicaments and B medicaments is the combination of thiophanate methyl and tebuconazole. The method can effectively control the damage of anthracnose and ring spot to the pear storage period, effectively reduce the using amount of the medicament, delay the drug resistance of the anti-medicament of anthracnose and ring spot, improve the fresh-keeping effect, reduce the storage loss, and have remarkable economic and social benefits.)

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

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 pear ring rot 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.

The verticillium pyricularis is also called as rough skin disease, the verticillium is a fungus, the sexual stage belongs to ascomycete subgenus, the asexual stage belongs to deuteromycete subgenus physiospora piricola Nose, (Macrophoma kuwatsukai Hara), and the verticillium pyricularis and apple verticillium are the same and can be infected mutually. After the fruit is infected with diseases, water stain-shaped, brown and nearly circular small spots occur on the surface of the fruit by taking a skin hole as a center, the small spots rapidly expand to the periphery to form ring-line type disease spots with dark brown and light brown alternating, the disease spots are not sunken and are soft rot, and small black spots are scattered in the disease part at the later stage, and the disease spots are conidiospores of germs. The diseased fruit quickly decays, brown juice can flow out, the shape of the fruit is still kept, and the fruit becomes stiff after dehydration and shrinkage. The latent period of the pathogenic bacteria is long, and symptoms are shown from the infection of young fruits to the near-mature period and the storage period. Is a serious disease in the storage period.

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 and ring rot of pears and application thereof. The invention provides the following technical scheme:

an antistaling agent for preventing and treating anthracnose and ring rot of pear is prepared from the antistaling agent A, B and bamboo vinegar, and the combination of chemical A and B is the combination of thiophanate-methyl and tebuconazole. 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 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 damage of anthracnose and ring spot to the pear storage period, effectively reduce the using amount of the medicament, delay the drug resistance of the anti-medicament of anthracnose and ring spot, improve the preservation effect, reduce the storage loss and have obvious economic and social benefits.

Drawings

FIG. 1: compound combination 2 fresh-keeping effect diagram for pear anthracnose

FIG. 2: compound combination 4 fresh-keeping effect picture for pear ring rot

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 and ring spot 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 and ring spot

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

Experiment number	Bamboo vinegar liquid	Agent A	Agent B
				1	1	1	1
2	1	2	3
				3	1	3	4
4	1	4	2
				5	2	1	2
6	2	2	4
				7	2	3	3
8	2	4	1
				9	3	1	3
10	3	2	1
				11	3	3	2
12	3	4	4
				13	4	1	4
14	4	2	2
				15	4	3	1
16	4	4	3

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 or tebuconazole.

TABLE 3 virulence of the combination (thiophanate-methyl, tebuconazole, bamboo vinegar) against anthracnose of pear

As can be seen from the table 3, the combined toxicity index CTC of the compounded combination (thiophanate methyl, tebuconazole and bamboo vinegar) is mostly more than 120, which indicates that the compounded agent has obvious synergistic effect.

TABLE 4 virulence of the combination (thiophanate-methyl, tebuconazole, bamboo vinegar) against pyricularia pyricularis

As can be seen from Table 4, the combined virulence indexes CTC of the compound are basically more than 120, which indicates that the compound 4 has obvious synergistic effect.

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 anthracnose and ring spot 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 2) thiophanate-methyl tebuconazole bamboo vinegar

876.5m1

43.8g

9.7g

/

70g

Suspending agent

Anthracnose

(compounding agent 4) thiophanate-methyl, tebuconazole and bamboo vinegar

854.5ml

74.3g

2.9g

/

68.3g

Suspending agent

Ring rot of the foot

Comparison of the medicaments: 43% tebuconazole emulsifiable concentrate and 70% thiophanate methyl wettable powder

Test results

Table 6: analysis table of in vivo experiment prevention and treatment effects of compound agent 2 and control group

As can be seen from Table 6, when the compound agent 2 is diluted according to 800, the control effect is equivalent to 800 times of 70 percent thiophanate-methyl and 4000 times of 43 percent tebuconazole; when diluted by 800 times, the control effect is slightly more than 5000 times of 43 percent tebuconazole.

The 800-time diluted liquid medicine of the compound agent 2 contains 0.055g of thiophanate methyl and 0.012g of tebuconazole 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, and the total amount of the active ingredients of the chemical pesticide is reduced by 92.4 percent; compared with a single dose of tebuconazole 4000-fold diluted liquid medicine, each liter of tebuconazole contains 0.107g, and the total amount of active ingredients of the chemical pesticide is reduced by 37.8 percent;

the 1000-time diluted liquid medicine of the compound agent 2 contains 0.044g of thiophanate-methyl and 0.010g of tebuconazole per liter, and compared with the 1200-time diluted liquid medicine of single-dose thiophanate-methyl, the per liter contains 0.583g of thiophanate-methyl, and the total amount of the active ingredients of the chemical pesticide is reduced by 90.8 percent; compared with 5000 times diluted medicinal liquid of single dose of tebuconazole, each liter of tebuconazole contains 0.107g, and the total amount of active ingredients of the chemical pesticide is reduced by 37.8 percent;

the compound agent 2 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 4 and control group

As can be seen from Table 7, the control effect of the compound agent 4 diluted according to 100 percent is equivalent to 800 times of 70 percent thiophanate-methyl and 4000 times of 43 percent tebuconazole; when diluted by 200 times, the control effect is slightly more than 5000 times of 43 percent tebuconazole.

The compound agent 4 is 100 times diluted liquid medicine, each liter contains 0.743g thiophanate-methyl and 0.029g tebuconazole, compared with the single agent thiophanate-methyl 800 times diluted liquid medicine, each liter contains 0.875g thiophanate-methyl, and the total amount of the effective components of the chemical pesticide is reduced by 11.8%;

the 200-time diluted liquid medicine of the compound agent 4 contains 0.372g of thiophanate-methyl and 0.015g of tebuconazole per liter, and compared with the 1200-time diluted liquid medicine of the single-dose thiophanate-methyl, the per liter contains 0.583g of thiophanate-methyl, and the total amount of the active ingredients of the chemical pesticide is reduced by 33.8 percent; the compound agent 4 has good prevention and treatment effect on the pear ring rot.

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 2 and not adding the bamboo vinegar liquid

Experimental group	Regression equation	Correlation coefficient	Co-toxicity coefficient
				Adding bamboo vinegar	Y＝2.15X-2.0638	0.9774	295.40
Bamboo vinegar liquid is not added	Y＝1.5582x-0.3274	0.97993	217.25

Bamboo vinegar EC50, thiophanate-methyl EC50, tebuconazole EC50 is 4: 3: 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 8, the co-toxicity coefficient of the pesticide is obviously improved by using the bamboo vinegar in the compound 2.

Table 9: the toxicity of the mixed solution 4 added with the bamboo vinegar solution is compared with that of the mixed solution without the bamboo vinegar solution on the anthracnose of pear

Experimental group	Regression equation	Correlation coefficient	Co-toxicity coefficient
				Adding bamboo vinegar	Y＝2.644X-3.5886	0.9929	226.47
Bamboo vinegar liquid is not added	Y＝1.4893x-0.1461	0.9681	140.59

Bamboo vinegar EC50, thiophanate-methyl EC50, tebuconazole EC50 is 1: 3

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 4.