CN107897198A - The complex composition and its application of Zhongshengmycin and eugenol - Google Patents

Abstract

The present invention relates to a kind of good with environment compatibility, efficiently, low toxicity, low-residual, the composite bactericide being made of Zhongshengmycin and eugenol, particularly for the application study of the fruit tree Major Diseases such as prevention ring rot of apple, apple anthracnose, fruit white rot of grape, bitter rot or anthracnose of grape.The weight ratio of Zhongshengmycin and two kinds of active ingredient of eugenol is 70 in complex composition:1~1:70, enhancing proportions 40:1~1:10, optimal proportion 10:1.It has outstanding performance in terms of the fruit tree Major Diseases such as application prevention ring rot of apple, apple anthracnose, fruit white rot of grape, bitter rot or anthracnose of grape.

Description

Compound composition of Zhongshengmycin and eugenol and its application

technical field

The invention relates to a compound composition of Zhongshengmycin and eugenol, especially the compound composition is used as a fungicide to prevent and treat important diseases of fruit trees such as apples and grapes, and belongs to the technical field of compound compositions of fungicides .

Background technique

Grape white rot, grape anthracnose, apple ring spot and apple anthracnose are one of the important diseases that seriously occur in the main fruit producing areas. At present, chemical fungicides are still the main means to prevent and control fruit tree diseases, but the long-term unreasonable large amount of chemical pesticides has led to problems such as increased control costs, excessive fruit residues, environmental pollution, and the development of drug resistance. The public's close attention to food safety and the growing call for global environmental protection require researchers to find effective and safe methods of prevention and control. Therefore, the development and utilization of pesticide resources that are compatible with the environment and harmless to humans are becoming increasingly popular in various countries. of favor.

Biological pesticides, also known as "biological pesticides", refer to pesticides developed using biological resources, whose active ingredients are naturally occurring, with low toxicity, no pollution, strong specificity, easy degradation, and the possibility of accumulating toxicity in the ecological environment small size and environmental compatibility. It can guarantee the increasing demand for organic food and green food, avoid or reduce the negative impact caused by the application of chemical pesticides, and has the advantages of making it difficult for pathogenic bacteria to develop drug resistance. It is an ideal type of pesticide.

Zhongshengmycin is an antibiotic produced by Streptomyces lilacaceae Hainan variety, which is an N-glycoside alkaline water-soluble substance. Its mechanism of action is mainly through inhibiting the synthesis of bacterial protein for bacteria, resulting in the death of bacteria; for fungi, it deforms filamentous hyphae, inhibits spore germination and can directly kill spores. Zhongshengmycin has contact and penetration effects, and has high activity against bacterial diseases and some fungal diseases of crops, and has a certain effect on increasing production. No crop pathogenic bacteria have been found to be resistant to it.

The botanical pesticide eugenol exists in clove oil, cinnamon leaf oil, cinnamon bark oil, camphor oil, nutmeg oil, etc., and can be separated from clove oil. Eugenol is absorbed by the leaves, stems, and roots of plants, and has an upward conduction function, is safe, environmentally friendly, and has no residues. It has rapid efficacy and long-lasting effect. After the diseased crops are sprayed, the fungal spores transform immediately and dissolve and disappear. In recent years, eugenol has attracted the attention of clinicians, scientific research circles and patients, and has also been used by scientific research institutes and manufacturers as a pesticide for promotion and application. It has not been reported that eugenol and Zhongshengmycin are used to control apple ring spot, apple anthracnose, grape white rot, and grape anthracnose.

Contents of the invention

The object of the present invention is to provide a single agent of Zhongshengmycin, eugenol and a compound composition thereof for preventing and treating important fruit tree diseases such as apple ring spot and grape white rot. The invention discusses the application of the compound composition of Zhongshengmycin and eugenol in the field of agriculture, especially for the research on the prevention and control of important diseases such as apple ring spot and grape white rot.

The present invention is achieved through the following technical solutions:

The compound composition of Zhongshengmycin and eugenol is special in that the weight ratio of the active ingredients Zhongshengmycin and eugenol is 70:1～1:70;

The compound composition of Zhongshengmycin and eugenol is further improved in that the synergistic weight ratio of the active ingredients Zhongshengmycin and eugenol is 40:1-1:10;

The compound composition of Zhongshengmycin and eugenol is further improved in that the optimal ratio of its active ingredients Zhongshengmycin and eugenol is 10:1, and the synergistic coefficient is 2.18;

The compound composition of Zhongshengmycin and eugenol, the total weight of the active ingredients Zhongshengmycin and eugenol in the whole compound composition accounts for 2.2% of the compound composition, and the balance is added by conventional excipients thing.

The compound composition of Zhongshengmycin and eugenol of the present invention is used for preventing and treating apple ringworm, apple anthracnose, grape white rot, and grape anthracnose.

The present invention also provides a method for using the compound composition of Zhongshengmycin and eugenol, which is special in that the compound composition of Zhongshengmycin and eugenol is diluted 600-800 times to prevent and treat Apple ringworm, apple anthracnose, grape white rot, and grape anthracnose have good control effects.

Zhongshengmycin and eugenol compound composition of the present invention has the following advantages:

1. It has a synergistic effect. The results of indoor toxicity test showed that Zhongshengmycin and eugenol showed synergistic effects on grape white rot fungus in the range of 40:1-1:10, and the synergistic effect of 10:1 was the most significant. The efficiency coefficient reaches 2.18.

2. The optimal ratio of field tank mix has outstanding control effect and reduces the dosage of preparations. The compound composition of 2.2% Zhongshengmycin·eugenol produced by our hospital was 600-800 times better in the field control of apple ring spot, apple anthracnose, grape white rot, and grape anthracnose. It is better than or equivalent to the control effect of single agent and control agent. At the same time, the amount of preparation is greatly reduced. Compared with tank mixing directly according to the concentration of the label, the compound composition reduces the amount of preparation by 33% and 50% respectively by 600 times and 800 times.

3. High security. Eugenol and Zhongshengmycin have low toxicity to humans and animals, and the compound composition prepared by our institute is lower than the content of active ingredients in a single dose on the market, which is safer for crops, the environment and fresh fruit eaters.

4. Reduce the risk of drug resistance of pathogenic bacteria. As mentioned in advantage 2, the amount of bacteriocin and eugenol in the compound composition is significantly lower than that of the commercial single-dose product, and the dosage of the preparation is significantly reduced when used in the field, which can greatly reduce pathogenic bacteria such as grape white rot Selection pressure on these two fungicides. In addition, Zhongshengmycin and eugenol have different mechanisms of action, so it is less likely to cause drug resistance in pathogenic bacteria when used in tank mixes.

Detailed ways

The present invention is described in detail with the following specific examples, but by no means limit the present invention. The percentages in all proportions are percentages by weight.

Example 1

Indoor toxicity test and compound ratio screening

1.1 Drugs to be tested

12% Zhongshengmycin wettable powder, product of Fujian Kaili Biological Products Co., Ltd., dissolved in water to make 1000mg/L mother liquor for later use.

0.3% eugenol soluble agent, product of Shandong Yijia Agrochemical Co., Ltd., dissolved in water to make 1000 mg/L mother liquor for later use.

1.2 Tested strains

Grape white rot (Coniothyrium diplodiella) was isolated from diseased fruits in the field, and the first-generation strain obtained by monospore culture and purification was stored in a refrigerator at 4°C for future use.

1.3 Instruments and equipment

Instruments and equipment mainly include electronic balances, ultra-clean benches, biological incubators, petri dishes, pipettes, etc. 1.4 Experimental method

The Mycelium growth rate test recommended by the FAO was used.

1.4.1 Drug compounding

The compound ratio of shengmycin and eugenol in table 1

Table 2 Compound drug dilution concentration gradient

1.4.2 Preparation of drug-containing tablet

According to the drug concentration gradient set in 1.4.1, add the drug to the quantitatively dissolved PDA medium to make a drug-containing plate, use the drug-free plate as a blank control, and repeat 3 times for each concentration.

1.4.3 Inoculation culture

Use a hole puncher with a diameter of 0.6 cm to punch out the plate along the edge of the cultured colony, use a sterile inoculation needle to invert the plate and transfer it to the plate, and place it at 25°C for constant temperature cultivation for 6 days.

1.4.4 Result survey

The expansion diameter of the bacterial colony was measured by the straightedge cross measurement method, and compared with the expansion diameter of the control plate without application of the drug, the inhibitory rate of the drug on the growth of the mycelium was calculated.

1.5 Statistical analysis

Regression analysis was performed according to the logarithmic value of each agent concentration and the probability value of the corresponding mycelia growth inhibition rate, and the toxicity regression equation, correlation coefficient (r) and EC ₅₀ of each agent were calculated.

In the determination of the combined toxicity of agents, the synergistic coefficient (SR) of the compound composition was calculated according to the Wadley method, and the combined effect type of the compound composition was evaluated. The synergistic effect of drug compounding is evaluated according to the synergistic coefficient (SR), that is, SR<0.5 is an antagonistic effect, 0.5≤SR≤1.5 is an additive effect, and SR>1.5 is a synergistic effect. Synergistic coefficient (SR) is calculated according to formula (1) and (2):

In the formula:

X ₁ - the theoretical value of EC ₅₀ of the compound composition, in mg/L;

P _A ---- the percentage content of A in the compound composition, the unit is percentage (%);

P _B ---- the percentage content of B in the compound composition, the unit is percentage (%);

A----EC ₅₀ value of A, the unit is mg/L;

B----EC ₅₀ value of B, the unit is mg/L.

In the formula:

SR--the synergistic coefficient of the compound composition;

X ₁ - the theoretical value of EC ₅₀ of the compound composition, in mg/L;

X ₂ ----The measured value of EC ₅₀ of the compound composition, the unit is mg/L.

1.6 Result Analysis

Through indoor toxicity experiments, the results showed that the two single-dose indoors showed good inhibitory effects on grape white rot fungus, with EC ₅₀ of 3.1942mg/L and 0.7089mg/L, respectively. Zhongshengmycin and eugenol showed a synergistic effect when the active ingredient ratio was 40:1-1:10, especially the ratio of 10:1 was the most significant, and its EC ₅₀ was 1.1098 mg/L, increasing The efficiency factor is 2.18. It shows that Zhongshengmycin·eugenol 10:1 compound fungicide has high fungicidal activity against grape white rot.

Screening of the optimal compound ratio of shengmycin and eugenol in table 3

Example 2

field plot experiment

According to the indoor compound ratio screening experiment, the optimal ratio of Zhongshengmycin:eugenol=10:1 was selected, and we prepared a 2.2% Zhongshengmycin·eugenol compound composition. According to the 13th, 121, 181 parts of the "Pesticide Field Efficacy Test Guidelines (2)" edited by the Pesticide Control Institute of the Ministry of Agriculture, 2.2% Zhongshengmycin·eugenol compound composition is designed to treat apple ringworm and apple anthracnose , Grape white rot, grape anthracnose field efficacy test.

2.1 Drugs to be tested

0.3% eugenol soluble solution 600 times, product of Shandong Yijia Agrochemical Co., Ltd.;

3% Zhongshengmycin wettable powder 600 times, product of Shandong Zhaofengnian Biotechnology Co., Ltd.;

2.2% Zhongshengmycin · Eugenol 600 times, 800 times, 1000 times, prepared by Yantai Academy of Agricultural Sciences.

2.2 Community design

A total of 7 treatments were set up in the experiment, and each treatment had 4 repetitions, and a total of 28 plots were set up, with 6 adult trees in each plot. The districts are arranged randomly. The pesticide application equipment adopts the Gongnong 26 high-pressure motorized sprayer, and sprays the medicine evenly on all the branches and leaves in the canopy until the water drips.

2.3 Investigation time and method

During the harvest period, select 3-4 plants in the center of each plot, investigate all the fruits, record the total number of fruits and the number of diseased fruits, and calculate the control effect based on the diseased fruit rate.

2.4 Calculation method of efficacy

2.5 Test results

2.2% Zhongshengmycin · Eugenol compound fungicide 600 and 800 times showed good control effects on grape white rot, the control effects were 88.57% and 85.30% respectively. Among them, the 5% level of the compound fungicide 600 times is significantly better than the control effect of 3% Zhongshengmycin 600 times and 0.3% eugenol 600 times of the two single agents, and it is comparable to 3% Zhongshengmycin WP 600 Times + 0.3% eugenol soluble solution 600 times tank mixed no significant difference. Although the control effect of the compound fungicide 800 times liquid is significantly lower than the above-mentioned tank mix at the 5% level, the control effect is also above 85%, and there is no significant difference with the 3% Zhongshengmycin WP 600 times, 5% Level significantly better than 0.3% eugenol soluble agent 600 times the control effect. It fully demonstrated that the 4.2% Zhongshengmycin·eugenol compound fungicide had a stronger fungicidal activity on grape white rot. (Table 4).

Table 4 Field efficacy of 2.2% Zhongshengmycin·eugenol compound composition in controlling grape white rot

The 600-fold and 800-fold control effects of the 2.2% Zhongshengmycin·eugenol compound fungicide on grape anthracnose were 87.62% and 85.30%, respectively. Among them, the 600 times control effect of compound fungicide is significantly better than 600 times of 3% Zhongshengmycin wettable powder and 600 times of 0.3% eugenol soluble solution at the level of 5%, and it is the same as that of 3% Zhongshengmycin WP 600 times. There was no significant difference between 600 times of shengmycin wettable powder + 600 times of 0.3% eugenol soluble solution. There is no significant difference between the 800-fold control effect of the compound fungicide and the 600-fold control effect of the 3% Zhongshengmycin WP, and it is significantly better than the 600-fold control effect of the 0.3% eugenol solution at the 5% level (Table 5).

Table 5 Field efficacy of 2.2% Zhongshengmycin·eugenol compound composition in controlling grape anthracnose

The field control results of 2.2% Zhongshengmycin·eugenol compound fungicide on apple ring spot showed that the control effect of the compound fungicide was 600 and 800 times better than that of 3% Zhongshengmycin at the level of 5%. Powder 600 times and 0.3% eugenol soluble agent 600 times the control effect of two single doses, and there is no significant difference with 3% Zhongshengmycin WP 600 times + 0.3% eugenol soluble agent 600 times tank mix (Table 6).

Table 6 Field efficacy of 2.2% Zhongshengmycin·eugenol compound composition in controlling apple ring spot

2.2% Zhongshengmycin · Eugenol compound fungicide has 600 times and 800 times control effects on apple anthracnose, which are 88.61% and 85.66% respectively, both of which are significantly better than 3% Zhongshengmycin in wettability at the level of 5%. Powder 600 times and 0.3% eugenol soluble agent 600 times the control effect of two single doses, and there is no significant difference with 3% Zhongshengmycin WP 600 times + 0.3% eugenol soluble agent 600 times tank mix .

Table 7 Field efficacy of 2.2% Zhongshengmycin·eugenol compound composition in controlling apple anthracnose

Example 3

safety test

The content of Zhongshengmycin and eugenol in the compound composition is less than that of commercially available products, so its toxicity to humans and animals and the safety of crops meet the requirements. The results of the toxicity test of the accredited institution show that the compound fungicide belongs to the slightly toxic pesticide:

Rat acute oral toxicity LD50♂>5000mg/kg,♀>5000mg/kg;

Rat acute dermal toxicity LD50♂>5000mg/kg,♀>5000mg/kg;

Rabbit eye irritation score is 0 (1h), after 48h it is 0 (1:100) dilution, no irritation to eyes;

Rabbit eye irritation score is 0 (4h), non-irritating to the skin.

The present invention has established the optimal ratio of the two pesticides for production use through indoor toxicity testing and field tests on the effects of the biogenic pesticides zhongshengmycin and eugenol on the main diseases of grapes and apples after they are used alone or compounded. In order to achieve the purpose of reducing pesticide environmental pollution and controlling the occurrence and development of bacterial resistance.

Claims (6)

1. The compound composition of Zhongshengmycin and eugenol, characterized in that the weight ratio of its active ingredients Zhongshengmycin and eugenol is 70:1～1:70. 2. According to the compound composition of Zhongshengmycin and eugenol according to claim 1, it is characterized in that the synergistic weight ratio of Zhongshengmycin and eugenol in its active ingredients is 40:1～1:10 . 3. According to the compound composition of Zhongshengmycin and eugenol according to claim 2, it is characterized in that the optimal ratio of its active ingredients Zhongshengmycin and eugenol is 10:1. 4. According to the composite composition of Zhongshengmycin and eugenol according to any one of claims 1-3, it is characterized in that the active ingredient Zhongshengmycin and eugenol in the whole composite composition The total weight accounts for 2.2% of the compound composition, and the balance is conventional auxiliary material additions. 5. The compound composition of Zhongshengmycin and eugenol according to any one of claims 1-3 is applied to the purposes of preventing and treating apple ringworm, apple anthracnose, grape white rot, and grape anthracnose. 6. according to the purposes described in claim 5, it is characterized in that, make 2.2% Zhongshengmycin·eugenol compound composition and dilute to 600-800 times to control apple ring spot, apple anthracnose, grape white rot disease, grape anthracnose.