CN107624767A - A kind of application of dipeptide compounds - Google Patents

Abstract

The invention discloses the application of a dipeptide compound. The dipeptides compound has a good inhibitory effect on the walnut anthracnose bacterium (Gloeosporium fructigenum), and the minimum inhibitory concentration (MIC) is 64 μ g/mL, which can be used to prepare agricultural fungicides; the structure of the dipeptides compound is as follows: (1) as shown. The compound is a natural product derived from microorganisms. Compared with traditional chemically synthesized fungicides, it is easy to degrade in the environment and easy to produce on a large scale.

Description

Application of a kind of dipeptide compound

technical field

The present invention relates to microorganism source fungicide, specifically the application of a kind of Dipeptides (dipeptide) compound.

Background technique

Walnuts have a long history of cultivation in my country. They are rich in protein and various trace elements such as calcium, phosphorus, and iron necessary for the human body. As the nutritive and medicinal value of walnut is gradually paid attention to, its planting area is getting larger and larger, but the occurrence of its diseases is more and more serious. As one of the main diseases, walnut anthracnose has the characteristics of long incubation period, short onset time, and strong explosiveness. It can cause a large number of walnut fruits to turn black in the late stage of ripening, and greatly reduce the yield of walnut. At present, the control methods mainly rely on chemically synthesized pesticides such as Bordeaux mixture, mancozeb, tuzet, carbendazim, and chlorothalonil. Although they can achieve certain control effects, they are also prone to environmental pollution and excessive pesticide residues. . Compared with traditional chemically synthesized pesticides, fungicides derived from microorganisms have the advantages of being safe to humans, animals and non-target organisms, good environmental compatibility, and less likely to develop resistance. Therefore, the development and application of microbial-derived pesticides is of great significance to human health, environmental protection and sustainable development of agriculture.

Dipeptides compounds shown in formula (1) are obtained by extracting and separating Trichoderma fermentation products (EGaro, CM Starks, PR Jensen, et al. Trichodermamides A and B, cytotoxic modified dipeptides from the marine-derived fungus Trichoderma virens[J] .J.Nat.Prod.2003,66,423-426), this document proves by experiment: the effect of Trichodermamides B anti-human colon cancer HCT-116 is remarkable in vitro, IC ₅₀ is 0.32 micrograms/ml, this document does not disclose Trichodermamides A and B other functions.

Contents of the invention

The purpose of the present invention is to provide a new application of dipeptide (Dipeptides) compounds. The invention proves through experiments that it has good inhibitory activity on the walnut anthracnose bacteria, and the minimum inhibitory concentration (MIC) is 64 μg/mL.

In order to achieve the above object, the technical scheme adopted in the present invention is: the application of a dipeptide (Dipeptides) compound, which has a good inhibitory effect on walnut anthracnose (Gloeosporium fructigenum), and can be used to prepare agricultural fungicides ; The structure of the Dipeptides compound is shown in formula (1).

The dosage form of the agricultural fungicide can be wettable powder, granule or sustained-release preparation, which is prepared by using the compound represented by formula (1) as an active ingredient and adding commonly used agricultural auxiliaries.

Advantages that the present invention has: the compound shown in formula (1) can be obtained by Trichoderma through fermentation, extraction and separation, and it has good inhibitory activity to walnut anthracnose bacteria, and the minimum inhibitory concentration (MIC) is 64 μ g/mL, shows that the Dipeptides compounds have good inhibitory effect on walnut anthracnose bacteria, and can be used as new pesticide components or lead compounds with bactericidal effect.

The Dipeptides compounds involved in the present invention can be fermented on a large scale by microorganisms, and have the characteristics of simple production process, short cycle, and low product cost; and they only contain four elements: carbon, hydrogen, oxygen, and nitrogen, and are easy to degrade in the environment. Human and animal toxicity is low.

detailed description

In order to clarify the understanding of the characteristics of the present invention, the present invention will be further elaborated below in conjunction with some non-limiting implementation examples.

Example 1:

The Dipeptides compound represented by formula (1) is a known compound of Dipeptides. The preparation process of the Dipeptides compound shown in formula (1) can refer to literature E Garo, CM Starks, PR Jensen, et al.Trichodermamides A and B, cytotoxic modified dipeptides from the marine-derived fungus Trichoderma virens[J].J.Nat .Prod.2003,66,423-426 described.

Embodiment 2: antibacterial activity test

The microdilution method was used to measure the antibacterial activity of the compound shown in the formula (1) against the walnut anthracnose bacterium (Gloeosporium fructigenum) ("From Natural Products to New Pesticide Creation-Principle and Method", Wu Wenjun, 2006, Chemical Industry Press).

1) Preparation of bacterial suspension

The test fungus Gloeosporium fructigenum was inoculated on the surface of PDA medium, and after culturing at 28°C for 72 hours, 2 mL of sterile 0.85% NaCl solution (containing 0.25% Tween-20) was drawn to wash the culture, and the culture was scraped off with a glass spatula. Colonies were gently scraped off. Pipette an appropriate amount of bacterial suspension into a sterile test tube and adjust to 0.5 McFarland turbidity (equivalent to 1.5×10 ⁸ CFU/mL) for later use.

2) Sample preparation

Take 1 mg of the sample to be tested (compound of formula (1)) and positive control (amphotericin B), dissolve in 100 μL DMSO, mix thoroughly, draw 50 μL of the sample solution into another centrifuge tube, and then add 50 μL DMSO , to obtain a sample solution whose concentration was halved. According to this method, 12 groups of sample solutions whose concentrations were successively halved were obtained.

3) MIC determination method

(1) Using aseptic operation, add the sample solutions of different concentrations after doubling dilution to a sterile 96-well polystyrene plate, add 5 μL of sample solution to each of the 1st to 12th wells, and add no sample Wells were used as blank control, and wells added with 5 μL DMSO solution were used as solvent control.

(2) After diluting 1000 times the indicator bacteria suspension equivalent to 0.5 McFarland turbidity with Sabouraud medium, take 95 μL and add them to the 96-well plate in sequence, so that the sample concentrations in the 1st to 12th wells are sequentially 512, 256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5 and 0.25 μg/mL. All the above samples were repeated three times. After gently shaking and mixing, the 96-well plate was sealed and placed in a biochemical incubator at 28°C for 72 hours.

(3) Use a microplate reader to measure the absorbance of each well at a wavelength of 600nm, and use the minimum sample concentration that completely inhibits the growth of the indicator bacteria in the small well as the MIC of the compound. (Note: The experiment is only meaningful when the indicator bacteria grow significantly in the negative control well; when a single hole jump occurs in the experiment, the highest drug concentration that inhibits the growth of the bacterial strain should be recorded; if multiple hole jumps occur, the results should not be reported, and the results should be Repeat experiment.)

The test result shows that the MIC value of the compound represented by formula (1) against walnut anthracnose (Gloeosporium fructigenum) is 64 μg/mL, and has good antibacterial activity.

The above experimental results prove that the compounds involved in the present invention have good antibacterial activity and can be used to prepare agricultural fungicides.

Claims (2)

1. application of the dipeptide compounds in terms of the disinfectant use in agriculture of preventing and treating walnut anthracnose is prepared shown in formula (one),

2. application as claimed in claim 1, it is characterized in that, the disinfectant use in agriculture is WP, granule or sustained release agent.