阅读说明：本技术 ε-聚赖氨酸的用途 (Use of epsilon-polylysine ) 是由 刘文波 秦春秀 缪卫国 林春花 李潇 于 2021-09-17 设计创作，主要内容包括：本发明公开了ε-聚赖氨酸在制备防治植物炭疽病菌病害的农药中的用途。本发明还公开了一种用于防治植物炭疽病菌病害的农药,其包括ε-聚赖氨酸30～50份。实施本发明,可有效防治植物炭疽病菌引起的病害。(The invention discloses an application of epsilon-polylysine in preparing a pesticide for preventing and treating plant anthracnose pathogen diseases. The invention also discloses a pesticide for preventing and treating plant anthracnose pathogen diseases, which comprises 30-50 parts of epsilon-polylysine. The invention can effectively prevent and control the diseases caused by the plant anthracnose pathogen.)

1. The application of epsilon-polylysine in preparing pesticides for preventing and treating plant anthracnose pathogen diseases.

2. The use according to claim 1, wherein the pesticide further comprises natamycin.

3. The use according to claim 2, wherein the weight ratio of natamycin to epsilon-polylysine in the pesticide is (1-2): 3.

4. the use according to claim 3, wherein the weight ratio of natamycin to epsilon-polylysine in the pesticide is 1: 2.

5. use according to any one of claims 1 to 4, wherein the plant is rubber, watermelon, mango or betel nut.

6. A pesticide for preventing and treating plant anthracnose pathogen diseases is characterized by comprising 30-50 parts of epsilon-polylysine.

7. The pesticide for preventing and treating plant anthracnose pathogen diseases according to claim 6, further comprising 15-25 parts of natamycin.

8. The pesticide for preventing and treating plant anthracnose pathogen diseases as claimed in claim 6, wherein the weight ratio of natamycin to epsilon-polylysine is (1-2): 3.

9. the pesticide for controlling plant anthracnose pathogen diseases according to claim 6, wherein the weight ratio of natamycin to epsilon-polylysine is 1: 2.

10. the pesticide for preventing and treating plant anthracnose pathogen diseases according to claim 6, which comprises the following components in parts by weight:

30-50 parts of epsilon-polylysine, 15-25 parts of natamycin, 0.1-5 parts of zein, 1-5 parts of citric acid, 2-10 parts of glycerol, 1-10 parts of tween and 20-30 parts of water;

the sum of the parts by weight of the components is 100 parts.

Technical Field

The invention relates to the field of pesticides, in particular to application of epsilon-polylysine in preparing a pesticide for preventing and treating plant anthracnose pathogen diseases.

Background

ε -polylysine (. epsilon. -PL) is a polymer of L-lysine, and its chemical structure is shown below. Epsilon-polylysine is light yellow powder, slightly bitter, stable to heat, strong in hygroscopicity, insoluble in organic solvents such as diethyl ether, ethanol and ethyl acetate, and easily soluble in water.

Epsilon-polylysine (epsilon-PL) can directly destroy the selectivity of cells and organelles to the survival of substances and depends on the complete energy metabolism of a membrane structure, so that the membrane of an endolysosol is broken, microorganisms are induced to generate autolysis, and finally, the cells are killed. The existing research shows that the epsilon-polylysine has better inhibiting effect on escherichia coli, staphylococcus aureus, lactobacillus bulgaricus, bacillus subtilis, salmonella, aspergillus flavus, candida albicans, aspergillus niger and the like. Therefore, the method is often applied to the preservation of foods such as dairy products, meat products, marine products and the like.

Disclosure of Invention

The invention aims to solve the technical problem of providing the application of epsilon-polylysine in preparing pesticides for preventing and treating plant anthracnose pathogen diseases.

The invention also aims to solve the technical problem of providing a pesticide for preventing and controlling plant anthracnose pathogen diseases.

In order to solve the technical problem, the invention provides the application of epsilon-polylysine in preparing pesticides for preventing and treating plant anthracnose pathogen. In the research process, the inventor accidentally finds that epsilon-polylysine also has an inhibiting effect on the anthrax bacteria. Thus, the above use is provided.

As an improvement of the technical scheme, the pesticide also comprises natamycin. Specifically, the inventor shows through research that the bacteriostatic concentration of the anthrax bacteria is high when only epsilon-polylysine is adopted; after the natamycin is introduced, the bacteriostatic concentration is obviously reduced, and the natamycin have obvious synergistic action.

As an improvement of the technical scheme, the weight ratio of natamycin to epsilon-polylysine in the pesticide is (1-2): 3. specifically, when the amount is within the range, the synergistic effect of the two is more remarkable.

As an improvement of the technical scheme, the weight ratio of natamycin to epsilon-polylysine in the pesticide is 1: 2.

as an improvement of the technical scheme, the plant is rubber, watermelon, mango or betel nut.

Correspondingly, the invention also discloses a pesticide for preventing and treating plant anthracnose pathogen diseases, which comprises 30-50 parts of epsilon-polylysine.

As an improvement of the technical scheme, the natamycin preservative also comprises 15-25 parts of natamycin.

As an improvement of the technical scheme, the weight ratio of natamycin to epsilon-polylysine is (1-2): 3.

as an improvement of the technical scheme, the weight ratio of natamycin to epsilon-polylysine is 1: 2.

as an improvement of the technical scheme, the paint comprises the following components in parts by weight:

30-50 parts of epsilon-polylysine, 15-25 parts of natamycin, 0.1-5 parts of zein, 1-5 parts of citric acid, 2-10 parts of glycerol, 1-10 parts of tween and 20-30 parts of water;

the sum of the parts by weight of the components is 100 parts. The auxiliary agents (emulsifier, surfactant, solvent, etc.) for the agricultural chemical in the present invention are not limited to the above embodiments. One skilled in the art can select suitable adjuvants according to the specific pesticide dosage form.

The preparation method of the pesticide comprises the following steps: mixing the above materials except Tween, adding Tween, and mixing.

The implementation of the invention has the following beneficial effects:

the invention provides an application of epsilon-polylysine in preparing a pesticide for preventing and treating plant anthracnose pathogen diseases. Based on the application, the pesticide for preventing and treating plant anthracnose pathogens can be prepared, so that the diseases caused by the anthracnose pathogens can be effectively prevented and treated. Furthermore, the natamycin is introduced, so that the antibacterial concentration of the epsilon-polylysine can be greatly reduced.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to specific embodiments.

Example 1 measurement of Effect of ε -polylysine (. epsilon. -PL) on inhibition of Colletotrichum rubber Tree anthrax

The experimental method comprises the following steps: sterilizing PDA culture medium at 121 deg.C for 20min, adding different mass concentrations of epsilon-PL and PDA culture medium into culture dish, mixing, and making the concentrations of epsilon-PL solution in each plate culture medium be 0, 50, 100, 200, 400, 600, and 800mg/L respectively. Taking fungus cakes with the diameter of 4mm from the edge of a rubber tree anthrax fungus colony cultured for 7 days by using a PDA culture medium, respectively inoculating the fungus cakes to the center of a plate culture medium, inoculating one fungus cake to each plate, and culturing for 7 days in an incubator at 28 ℃ with the hypha facing downwards. Each group was repeated 4 times. Wherein, the control group is the solution with the concentration of the epsilon-PL of 0, and the other groups are the test groups.

After the experiment was completed, the diameter D of each colony group was measured_iAnd calculating the inhibition ratio omega_i. Then calculating the toxicity equation and half-maximum effect concentration EC of the epsilon-PL on the growth of the colletotrichum gloeosporioides mycelium by taking the logarithm value of the epsilon-PL concentration as the abscissa and the inhibition rate as the ordinate₅₀。

Wherein, the inhibition rate is calculated by the following formula group:

Δd_i＝D_i-d_i

wherein d is_iColony diameter for group i before experiment, D_iColony diameter, Δ d, of group i after experiment_iGrowth diameter for group i colonies, Δ d_cThe colony diameter is increased for the control group.

The experimental results are as follows: the inhibition effect on the growth of the colletotrichum gloeosporioides mycelium is gradually enhanced along with the increase of the concentration of epsilon-PL. After 2 days of culture, compared with a control group, the epsilon-PL treatment group can obviously inhibit the growth of rubber tree anthrax mycelium (P is less than 0.05), and when the addition amount of epsilon-PL is more than or equal to 600mg/L, the rubber tree anthrax basically does not grow. After 7 days of culture, the mycelium of the control group grows rapidly, and the diameter of the colony reaches 75.53 +/-0.15 mm. After being treated by epsilon-PL of 800mg/L, the growth of mycelium can be inhibited by more than 99.50 percent. The epsilon-PL treatment has an inhibiting effect on the growth of the mycelium of the colletotrichum gloeosporioides, and the regression equation of the toxicity is that Y is 1.5672X +1.2145, R²0.9964, half maximal effect concentration EC₅₀It was 263.027 mg/L.

Example 2 measurement of Effect of Epsilon-polylysine (. epsilon. -PL) in combination with Natamycin on inhibition of Colletotrichum heverae

2.1 determination of Effect of Natamycin on inhibition of rubber Tree anthrax

And (3) sterilizing the PDA culture medium for 20min at 121 ℃ in an autoclave, taking out, adding natamycin with different mass concentrations and PDA culture medium into a culture dish, and fully mixing to ensure that the concentration of the natamycin solution in each plate culture medium is 0, 5, 10, 15, 20, 25 and 30mg/L respectively for later use. Taking fungus cakes with the diameter of 4mm from the edge of a rubber tree anthrax fungus colony cultured for 7 days by using a PDA culture medium, respectively inoculating the fungus cakes to the center of a plate culture medium, inoculating one fungus cake to each plate, and culturing for 7 days in an incubator at 28 ℃ with the hypha facing downwards. Each group was repeated 4 times. Wherein, the natamycin solution with the concentration of 0 is a control group, and the others are test groups.

After the experiment was completed, the diameter D of each colony group was measured_iAnd calculating the inhibition ratio omega_i. Then calculating a toxicity equation and half-maximum effect concentration EC of the natamycin on growth of rubber tree anthrax mycelium by taking the logarithmic value of the natamycin concentration as the abscissa and the inhibition rate as the ordinate₅₀。

Wherein, the inhibition rate is calculated by the following formula group:

Δd_i＝D_i-d_i

wherein d is_iColony diameter for group i before experiment, D_iColony diameter, Δ d, of group i after experiment_iGrowth diameter for group i colonies, Δ d_cThe colony diameter is increased for the control group.

The experimental results are as follows: with the increase of the concentration of the natamycin, the inhibition effect on the growth of the rubber tree anthrax mycelium is gradually enhanced. After 2 days of culture, the natamycin treatment group can obviously inhibit the growth of rubber tree anthrax mycelium (P is less than 0.05) compared with the control group, and when the addition of natamycin is more than or equal to 25mg/L, the rubber tree anthrax basically does not grow. After 7 days of culture, control mycelia were grownRapidly, the colony diameter reached 85.67. + -. 0.13 mm. After being treated by 30mg/L natamycin, the growth of mycelium can be inhibited by more than 99.50 percent. The natamycin treatment has an inhibiting effect on the growth of the mycelium of the colletotrichum gloeosporioides, and the regression equation of the toxicity is that Y is 4.8764X +0.1438, R²0.9872, half maximal effect concentration EC₅₀It was 9.91 mg/L.

2.2 measurement of Effect of suppressing Anthrax of rubber Tree by combination of Epsilon-polylysine (Epsilon-PL) and Natamycin

The experimental method comprises the following steps: the following eight formulas are respectively set:

mixture A, epsilon-polylysine: natamycin ═ 6:4

Mixture B, epsilon-polylysine: natamycin ═ 6:3

Mixture C, epsilon-polylysine: natamycin ═ 6:2

Mixture D, epsilon-polylysine: natamycin ═ 6:1

Mixture E, epsilon-polylysine: natamycin ═ 1:6

Mixture F, epsilon-polylysine: natamycin ═ 2:6

Mixture G, epsilon-polylysine: natamycin ═ 3:6

Mixture H, epsilon-polylysine: natamycin ═ 4: 6.

And (3) sterilizing the PDA culture medium for 20min at 121 ℃ in an autoclave, taking out, adding the epsilon-PL, the natamycin and the PDA culture medium with different mass concentration ratios into a culture dish, and fully and uniformly mixing to ensure that the total concentration of the epsilon-PL and natamycin mixed ratio solution in each plate culture medium is 0, 10, 20, 30, 40 and 50mg/L respectively for later use. Taking fungus cakes with the diameter of 4mm from the edge of a rubber tree anthrax fungus colony cultured for 7 days by using a PDA culture medium, respectively inoculating the fungus cakes to the center of a plate culture medium, inoculating one fungus cake to each plate, and culturing for 7 days in an incubator at 28 ℃ with the hypha facing downwards. Each group was repeated 4 times. Wherein, the control group is the solution with the concentration of the epsilon-PL and the natamycin being 0, and the other groups are the test groups.

After the experiment was completed, the diameter D of each colony group was measured_iAnd calculating the inhibition ratio omega_i. Further taking the logarithm value of the total concentration of the epsilon-PL and the natamycin as the abscissa,the inhibition rate is the ordinate, and the toxicity equation and the half-maximum effect concentration EC are calculated₅₀。

In addition, the synergy coefficient SR is calculated according to the following formula group, and concretely, when the SR is more than or equal to 1.5, the synergy is shown; SR < 0.5 shows antagonistic action; an additive effect is that SR is more than 0.5 and less than 1.5.

Wherein, w_iIs the weight percentage of the ith component in the mixture; EC50_iHalf maximal effective concentration of component i alone, E_obHalf-maximal effect concentration of the mixture, E, for practical observation_thThe half-maximal effect concentration of the mixture is calculated.

Wherein, the inhibition rate is calculated by the following formula group:

Δd_i＝D_i-d_i

wherein d is_iColony diameter for group i before experiment, D_iColony diameter, Δ d, of group i after experiment_iGrowth diameter for group i colonies, Δ d_cThe colony diameter is increased for the control group.

The results of the experiments are shown in the following table:

as can be seen from the table, the combination of epsilon-polylysine and natamycin can effectively reduce the half-maximal effect concentration EC₅₀. Specifically, compared with the single epsilon-polylysine, the reduction range of the epsilon-polylysine reaches 21.3 to 91.4 times. The two have good synergistic effect. Especially when the weight ratio of natamycin to epsilon-polylysine is (1-2): 3, the synergistic effect is more obvious.

Example 3

The embodiment provides a pesticide for preventing and controlling plant anthracnose pathogen diseases, which comprises the following components:

40 parts of epsilon-polylysine, 20 parts of natamycin, 1 part of zein, 2 parts of citric acid, 5 parts of glycerol, 803 parts of tween and 29 parts of water.

The preparation method comprises the following steps:

mixing the above materials except Tween 80, adding Tween, and mixing.

EXAMPLE 4 mango field test

The test is arranged in a test base of plant protection institute of Hainan university, delirium, Hainan, the test land is a flat land parcel, and the soil is sandy soil. The cultivation management is uniformly standardized according to pesticide field pesticide effect tests.

1. Test material

Medicament: the pesticide of example 3 was used. The contrast agent is 30% pyraclostrobin suspending agent, is diluted by 800 times for use, and is sprayed with clear water as a blank contrast.

Pathogen: colletotrichum gloeosporioides

Host plant: mango (mango)

2. Test method

The area of the test cell is 40 square meters, and 6 mango trees are repeated in each cell for four times. The first water adding and spraying is carried out at the initial stage of the anthracnose of the mango leaves (sporadic attack), the spraying is carried out for 2 times at intervals of 5-7 days, and the liquid medicine spraying amount is 90L/mu.

3. Investigation method

Randomly sampling every cell, investigating 3 plants at each point, randomly sampling and investigating 3 plants at each cell in the shoot stage, sampling every plant according to east, west, south, north and middle five points, investigating all leaves of 2 shoots at each point, recording and investigating the total leaf number and the leaf number of each grade of disease, and then calculating the disease index and the prevention and treatment effect.

The investigation morbidity grading standard refers to the industrial standard for preventing and treating mango anthracnose, and is divided into 6 grades:

level 0: no disease spots;

level 1: the lesion area accounts for less than 5% of the whole leaf area;

and 3, level: the lesion area accounts for 6 to 15 percent of the whole leaf area;

and 5, stage: the lesion area accounts for 16 to 25 percent of the whole leaf area;

and 7, stage: the lesion area accounts for 26-50% of the whole leaf area;

and 9, stage: the lesion area accounts for more than 51% of the whole leaf area.

4. Drug effect calculation method

5. Test results

The disease index and the prevention effect of each treatment are shown in the following table:

the test data were subjected to a biometric analysis using the duncan's new complex range method (DMRT).

Example 5 watermelon field test

The test is arranged in the Fugu Foluruo county of Ledong province in Hainan, the land for testing is a plain land parcel, and the soil is sandy soil. The cultivation management is uniformly standardized according to pesticide field pesticide effect tests.

1. Test material

Medicament: the pesticide of example 3 was used. The contrast agent is 30% pyraclostrobin suspending agent, is diluted by 800 times for use, and is sprayed with clear water as a blank contrast.

Pathogen: watermelon anthracnose (Colletetrotrichum lagenarium)

Host plant: watermelon

2. Test method

The area of the test plot is 40 square meters, 1500 watermelons are planted per mu, and the test is repeated for four times. The first water adding and spraying is carried out at the initial stage (sporadic onset) of the anthracnose of the watermelon leaves for 2 times at intervals of 5-7 days, and the spraying liquid medicine amount is 60L/mu.

3. Investigation method

Sampling at five random points in each cell, investigating 3 plants at each point, investigating 5-10 leaves from top to bottom, recording disease grade numbers respectively, and calculating disease index and prevention and treatment effect.

The investigation disease grading standard is divided into 6 grades according to the industry standard of preventing and treating watermelon anthracnose:

level 0: no disease spots;

level 1: the lesion area accounts for less than 5% of the whole leaf area;

and 3, level: the lesion area accounts for 6 to 15 percent of the whole leaf area;

and 5, stage: the lesion area accounts for 16 to 25 percent of the whole leaf area;

and 7, stage: the lesion area accounts for 26-50% of the whole leaf area;

and 9, stage: the lesion area accounts for more than 51% of the whole leaf area.

4. Drug effect calculation method

5. Test results

The disease index and the prevention effect of each treatment are shown in the following table:

the test data were subjected to a biometric analysis using the duncan's new complex range method (DMRT).

Example 6 field test of betel nut

The test is arranged in a test base of plant protection academy of academic Press in the university of Hainan, Hainan province, the test land is a plain land, and the soil is sandy soil. The cultivation management is uniformly standardized according to pesticide field pesticide effect tests.

1. Test material

Medicament: the pesticide of example 3 was used. The contrast agent is 30% pyraclostrobin suspending agent, is diluted by 800 times for use, and is sprayed with clear water as a blank contrast.

Pathogen: colletotrichum gloeosporioides

Host plant: betel nut

2. Test method

The area of the test cell is 20 square meters, 6 areca-nuts are planted in each cell, and the test is repeated for four times. The first water adding and spraying is carried out at the initial stage of betel nut leaf anthracnose (sporadic onset), the continuous spraying is carried out for 2 times at intervals of 5-7 days, and the liquid medicine spraying amount is 90L/mu.

3. Investigation method

Randomly sampling in each cell, investigating 3 plants at each point, randomly sampling and investigating 3 plants in each cell in the shoot period, investigating all leaves of each betel nut tree, recording and investigating the total leaf number and the leaf number of each disease, and then calculating the disease index and the prevention and treatment effect.

The investigation disease grading standard is divided into 6 grades according to the industrial standard for preventing and treating areca anthracnose:

level 0: no disease spots;

level 1: the lesion area accounts for less than 5% of the whole leaf area;

and 3, level: the lesion area accounts for 6 to 15 percent of the whole leaf area;

and 5, stage: the lesion area accounts for 16 to 25 percent of the whole leaf area;

and 7, stage: the lesion area accounts for 26-50% of the whole leaf area;

and 9, stage: the lesion area accounts for more than 51% of the whole leaf area.

4. Drug effect calculation method

5. Test results

The disease index and the prevention effect of each treatment are shown in the following table:

the test data were subjected to a biometric analysis using the duncan's new complex range method (DMRT).

From examples 4-6, it can be seen that the pesticide of the invention has good control effect on anthracnose pathogen.

While the foregoing is directed to the preferred embodiment of the present invention, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention.