N-acetylglycine copper antibacterial agent

Abstract

The invention belongs to the technical field of antibacterial agents, particularly relates to preparation and application of a copper N-acetylglycinate antibacterial agent, and aims to provide preparation and application of an antibacterial agent taking copper N-acetylglycinate as an active component. The copper N-acetylglycine antibacterial agent prepared by the invention has the advantages of strong antibacterial effect, wide antibacterial spectrum and the like. Biological activity experiment determination shows that the prepared N-acetylglycine copper antibacterial agent has an efficient antibacterial effect on botanical pathogenic bacteria such as pestalotiopsis, strawberry fusarium wilt, rhizoctonia solani, fusarium moniliforme and pseudomonas solanacearum, and can be used as an agricultural bactericide to be applied to agricultural production.

Description

Technical Field

[0001] The invention belongs to the technical field of antibacterial agents, and specifically relates to an N-acetylglycine copper antibacterial agent, and aims to provide an agricultural antibacterial agent with strong antibacterial effect, broad antibacterial spectrum and safety to plants. Background Art

[0002] As early as 1807, B. Prevost discovered that copper sulfate had bactericidal activity. In 1882, Millarder A discovered that the solution of copper sulfate mixed with lime water, namely Bordeaux mixture, also had bactericidal activity and was safer. Copper preparations began to develop from then on. Common copper preparations are divided into two categories: inorganic copper fungicides and organic copper fungicides.

[0003] Inorganic copper fungicides are copper-based pesticides processed and formulated from natural mineral raw materials (such as copper sulfate, etc.). Bordeaux mixture, copper oxychloride, cuprous oxide, copper hydroxide, basic copper sulfate, etc. are all inorganic copper fungicides. Inorganic copper fungicides have a lot of disadvantages. Most fungicides have a narrow fungicide spectrum and are prone to allergies to individual varieties of crops. Their liquid medicines are alkaline and generally cannot be mixed with acidic pesticides. When used during the sensitive period of crops, they are prone to phytotoxicity and can easily induce the production of mites. The various disadvantages of inorganic copper fungicides have promoted the development and rapid growth of organic copper fungicides.

[0004] Organic copper fungicides are copper pesticides made by synthesizing copper ions and organic groups using organic chemical synthesis methods. For example, copper quinoline, copper thiophanate-methyl, copper chlorpyrifos, copper nonathiapiprolin, etc. are all organic copper fungicides. The copper ions and organic groups of organic copper fungicides can play a synergistic role and use a dual mechanism to kill bacteria. They have a wide bactericidal spectrum, low residue, low pollution, safety and reliability, good compatibility and affinity with the environment, and can be mixed with other pesticides. Therefore, compared with inorganic copper fungicides, organic copper fungicides with low copper content, low residue, and less environmental pollution are more in line with modern green requirements.

[0005] Over the years, people have conducted extensive research on the antibacterial mechanism of copper and proposed a variety of antibacterial mechanisms. Multiple antibacterial mechanisms work together, that is, multi-point bactericidal effects, so copper fungicides are not prone to resistance. The specific bactericidal mechanisms of copper include: (1) copper preparations release copper ions (Cu 2+ ), Cu 2+ Positively charged, bacterial cell membrane negatively charged, Cu 2+ It can firmly adsorb on the cell membrane, destroy the integrity of the cell membrane, cause cell rupture, cytoplasm outflow, hinder microbial reproduction, and ultimately lead to microbial death; (2) Cu 2+After passing through the cell membrane and entering the cell, it can chelate with amino acids with -SH groups in fungi or bacteria, causing protein denaturation and destroying enzymes that promote metabolism in the cell, thereby hindering and inhibiting the metabolism of microorganisms; (3) Cu 2+ It can directly affect the stability of microbial DNA and RNA, interfere with the replication and expression of microbial genetic material, and lead to mutation and death of microbial strains; (4) Cu 2+ It promotes the formation of ROS such as hydrogen peroxide and hydroxyl free radicals, causing various harms to cells, such as damaging the DNA of microorganisms, interfering with mitochondrial function, and destroying the integrity of cell membranes.

[0006] Copper N-acetylglycinate, C4H7NO3, CAS No. 543-24-8, is a white crystalline powder. Copper N-acetylglycinate is easily soluble in alcohol, soluble in water, slightly soluble in acetone and chloroform. It is an important fine organic chemical intermediate and is widely used in medicine, pesticides, chemical industry and other fields.

[0007] At present, there are no reports on the preparation of N-acetylglycine copper antibacterial agent and its application in agriculture. Summary of the invention

[0008] The purpose of the present invention is to provide a preparation and application of an antibacterial agent with copper N-acetylglycinate as the main active ingredient, aiming to provide an agricultural antibacterial agent with strong antibacterial effect, broad antibacterial spectrum and safety to plants, which is mainly used for preventing and controlling plant diseases.

[0009] In order to achieve the purpose of the present invention, the technical solution of the present invention is:

[0010] The antibacterial agent of the present invention uses copper N-acetylglycinate as an active ingredient and is used to prevent and control plant diseases;

[0011] The antibacterial agent of the present invention can be in the form of an aqueous solution, a soluble powder, a water-dispersible granule, or a soluble liquid.

[0012] Preferably, the antibacterial agent of the present invention is in the form of an aqueous solution;

[0013] The aqueous solution of the present invention uses copper N-acetylglycinate as an active ingredient and is prepared from the following components in percentage by mass: 0.5% to 50% copper N-acetylglycinate, 0.5% to 10% wetting and dispersing agent, 0.5% to 10% thickener, 0.5% to 10% antifreeze agent, and the balance is supplemented by deionized water;

[0014] The aqueous solution of the present invention, wherein the auxiliary component comprises one or more of a wetting and dispersing agent, a thickening agent, and an antifreeze agent;

[0015] The aqueous solution of the present invention adopts the following auxiliary agents: the wetting and dispersing agent is one or more of Tween-20, Tween-80, isooctyl alcohol polyoxyethylene ether, isotridecanol polyoxyethylene ether, alkylphenol polyoxyethylene ether phosphate, fatty alcohol polyoxyethylene ether phosphate, octylphenol polyoxyethylene ether phosphate, and polycarboxylic acid sodium salt; the thickener is one or more of xanthan gum, hydroxyethyl cellulose, hydroxymethyl cellulose, and magnesium aluminum silicate; the antifreeze agent is one or more of ethylene glycol, propylene glycol, glycerol, diethylene glycol, and triethylene glycol;

[0016] The antibacterial agent of the present invention is used for preventing and controlling plant diseases.

[0017] Attached pictures:

[0018] Figure 1 :Results of indoor toxicity test of Trichosporon pseudodiscus;

[0019] Figure 2 :Results of indoor toxicity test of strawberry wilt pathogen;

[0020] Figure 3 :Results of indoor toxicity test of Rhizoctonia solani;

[0021] Figure 4 :Results of indoor toxicity test of Fusarium moniliforme;

[0022] Figure 5 : Diagram of the diameter of the inhibition zone of tomato bacterial wilt pathogen. DETAILED DESCRIPTION

[0023] The present invention will be further described below in conjunction with specific examples, but the protection scope of the invention is not limited thereto. It should be understood that these examples are preferred embodiments of the present invention and are only used for specific illustration. Based on the above discussion and these examples, those skilled in the art can determine the essential features of the present invention, and can make various changes and modifications to the present invention without departing from the purpose and technical scope of the present invention to adapt it to various uses and conditions.

[0024] The "copper acetate" described in the embodiments of the present invention was purchased from Shanghai McLean Biochemical Technology Co., Ltd.; the "N-acetylglycine" was purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd.; the "15% copper chelate aqueous solution" was purchased from Shanxi Kexing Pesticide Liquid Fertilizer Co., Ltd., and the pesticide registration number is PD20111191; the "20% morphoguanidine·copper sulfate aqueous solution" was purchased from Henan Guofeng Jianyuan Agricultural Technology Co., Ltd., and the pesticide registration number is PD20100185; the "21.4% copper chelate·copper citric acid aqueous solution" was purchased from Shanxi Yonghe Lifeng Biotechnology Co., Ltd., and the pesticide registration number is PD20110220.

[0025] The test agent used in the embodiment of the present invention is 12% N-acetylglycinate copper aqueous solution, and the control agents used are 15% ammonia copper aqueous solution, 20% morphoguanidine·copper sulfate aqueous solution, and 21.4% copper chelate·copper citrate aqueous solution.

[0026] Example 1: Preparation of 12% N-acetylglycine copper aqueous solution

[0027] 1. Formula (parts by weight):

[0028] 12 parts of N-acetylglycinate copper, 3 parts of Tween-80, 0.5 parts of hydroxyethyl cellulose, 5 parts of ethylene glycol, and the balance is made up with deionized water.

[0029] 2. Preparation method:

[0030] According to the formula, take N-acetylglycinate copper, Tween-80, hydroxyethyl cellulose, and ethylene glycol, and make up the balance with deionized water. Add the above materials to a high shear emulsifier and mix them evenly to obtain a 12% N-acetylglycinate copper aqueous solution.

[0031] Example 2: Indoor toxicity test determination

[0032] Test strains: Trichosporon pseudodiscus, Fusarium wilt, Rhizoctonia solani, Fusarium moniliforme

[0033] Test method: Indoor toxicity test of the above strains was carried out by mycelium growth rate method

[0034] Experimental steps:

[0035] Use 12% N-acetylglycinate copper aqueous solution as the test solution, and use 15% chelate copper aqueous solution, 20% morphoguanidine·copper sulfate aqueous solution, and 21.4% chelate copper·citrate copper aqueous solution as the control solution. Dilute the above four solutions with water to the same copper content, and then prepare them into solutions with concentrations of 1600μg / mL, 800μg / mL, 400μg / mL, 200μg / mL, 100μg / mL, and 50μg / mL, respectively, for use.

[0036] Dilute the test solution and control solution of different concentrations with PDA agar medium cooled to 55℃±1℃ at a ratio of 1:9, shake well and pour into the plate medium, about 20mL per dish, to prepare the test plate medium for use. Mix sterile water and PDA agar medium according to the above operation to prepare the plate medium as the blank control medium.

[0037] Take 1mL of bacterial solution in logarithmic growth phase, such as strawberry wilt pathogen, inoculate it into ordinary PDA plate culture medium, seal it, and place it in an electric constant temperature incubator for 72h. Take a 5mm diameter bacterial cake at the edge of the colony, insert it into the test plate culture medium, ensure that the side containing the colony is in full contact with the test plate culture medium, seal it, and place it in an electric constant temperature incubator for 5d. Use the cross method to measure the mycelium diameter. Perform the experiment in parallel 3 times and take the average value. The same operation method is used for Pseudomonas aeruginosa, Rhizoctonia solani, and Fusarium moniliforme. Calculate the mycelium growth inhibition rate according to the following formula.

[0038]

[0039] The indoor toxicity curves of the four aqueous solutions against the four tested bacterial species are shown in the attached figure. Figure 2-5 shown.

[0040] According to the control effects of the four aqueous solutions on the four tested bacterial species, the 12% N-acetylglycine copper aqueous solution of the present invention has a significantly better antibacterial effect on Trichosporon, Fusarium wilt, Rhizoctonia solani and Fusarium moniliforme than 15% chlorammine copper aqueous solution, 20% morphoguanidine·copper sulfate aqueous solution and 21.4% chlorammonium·copper citrate aqueous solution.

[0041] Example 3: Inhibition zone determination

[0042] Test strain: Pseudomonas solanacearum

[0043] Test method: punching method

[0044] Experimental steps:

[0045] Use 12% N-acetylglycinate copper aqueous solution as the test solution, and use 15% ammonia copper aqueous solution, 20% morphoguanidine·copper sulfate aqueous solution, and 21.4% chelate·citrate copper aqueous solution as the control solution. Dilute the above four solutions with water to the same concentration of active ingredients based on the weight concentration of copper element, and prepare them into solutions with concentrations of 1600μg / mL, 800μg / mL, 400μg / mL, 200μg / mL, 100μg / mL, and 50μg / mL, respectively, for use.

[0046] Pipette 1mL of Pseudomonas solanacearum bacterial solution into a 200mL NA agar medium flask, shake well and pour into the plate culture medium, about 20mL per dish. After it is completely solidified, use a sterile hole puncher to punch 3 equally spaced holes with a diameter of 5mm on the plate. The size and depth of each hole should be kept consistent as much as possible. Pipette 100μL of different concentrations of test agents and control agents into the holes, add the same concentration of solution to the same plate, seal it, put it in an electric constant temperature incubator, select a suitable culture temperature and culture for 24 hours, and use the cross method to measure the colony diameter. The results of the inhibition zone determination of the above test solution and control solution against tomato bacterial wilt pathogen are shown in the attached figure. Figure 5shown.

[0047] The degree of inhibition of the above-mentioned agents on Pseudomonas solanacearum was determined according to the antibacterial activity determination standard.

[0048] The antibacterial activity determination standard is: the diameter of the inhibition zone is less than or equal to 7mm, which means no antibacterial effect; the diameter of the inhibition zone is greater than 7mm and less than or equal to 10mm, which means low antibacterial effect; the diameter of the inhibition zone is greater than 10mm and less than or equal to 20mm, which means moderate antibacterial effect; the diameter of the inhibition zone is greater than 20mm, which means high antibacterial effect.

[0049] The results showed that all four aqueous solutions had antibacterial effects against Pseudomonas solanacearum, and the antibacterial ability of 12% N-acetylglycine copper aqueous solution was significantly stronger than that of 15% chelate copper aqueous solution, 20% morphoguanidine·copper sulfate aqueous solution and 21.4% chelate copper·citrate copper aqueous solution.

Claims

1. A copper N-acetylglycinate antibacterial agent, characterized in that: The preparation of the invention uses copper N-acetylglycinate as an active ingredient and is used for preventing and controlling plant diseases.

2. An antibacterial agent according to claim 1, characterized in that: The dosage form of the preparation of the present invention can be aqueous solution, soluble powder, water dispersible granule, soluble liquid.

3. An antibacterial agent according to claim 1, characterized in that: The dosage form of the preparation of the present invention is specifically an aqueous solution.

4. The aqueous solution according to claim 3, characterized in that: The preparation of the present invention is prepared from copper N-acetylglycinate and auxiliary agents: 0.5% to 50% copper N-acetylglycinate, 0.5% to 10% wetting and dispersing agent, 0.5% to 10% thickener, 0.5% to 10% antifreeze agent, and the balance is supplemented with deionized water; The above materials are added into an open bottle and subjected to high shearing to be refined and uniformly broken down to obtain N-hydroxymethylglycinate copper aqueous solution.

5. The aqueous solution according to claim 3, characterized in that: The auxiliary agent of the present invention is selected from one or more of a wetting and dispersing agent, a thickener, an antifreeze agent, a defoaming agent, a stabilizer, and a solubilizing agent.

6. The aqueous solution according to claim 4, characterized in that: The wetting and dispersing agent in the aqueous solution of the present invention is one or more of Tween-20, Tween-80, isooctyl alcohol polyoxyethylene ether, isotridecanol polyoxyethylene ether, alkylphenol polyoxyethylene ether phosphate, fatty alcohol polyoxyethylene ether phosphate, octylphenol polyoxyethylene ether phosphate, and polycarboxylic acid sodium salt; the thickener is one or more of xanthan gum, hydroxyethyl cellulose, hydroxymethyl cellulose, and magnesium aluminum silicate; the antifreeze agent is one or more of ethylene glycol, propylene glycol, glycerol, diethylene glycol, and triethylene glycol.

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