阅读说明：本技术 和厚朴酚在制备天然环境友好型农药中的用途 (Application of honokiol in preparation of natural environment-friendly pesticide ) 是由 荆常亮 张成省 刘明宏 汪娴娴 李义强 梁晨 邹平 袁源 赵栋霖 于 2020-12-08 设计创作，主要内容包括：本发明涉及一种和厚朴酚在制备天然环境友好型农药中的用途,属于和厚朴酚的新应用技术领域。本发明提供了一种和厚朴酚在制备天然环境友好型农药中的用途。所述的和厚朴酚分子量为266.33,分子式为C_(18)H_(18)0_2。所述的用途,用于防治作物细菌病害和真菌病害。本发明提供的和厚朴酚在制备天然环境友好型农药中的用途,为作物病害的防治提供了一种新的安全有效的物质,提高了防治效果,降低了施药成本。(The invention relates to application of honokiol in preparation of natural environment-friendly pesticides, and belongs to the technical field of new application of honokiol. The invention provides an application of honokiol in preparation of natural environment-friendly pesticides. The molecular weight of the honokiol is 266.33, and the molecular formula is C 18 H 18 0 2 . The application is used for preventing and treating crop bacterial diseases and fungal diseases. The application of the honokiol in preparing the natural environment-friendly pesticide provided by the invention provides a new safe and effective substance for preventing and treating crop diseases, improves the prevention and treatment effect and reduces the pesticide application cost.)

1. Application of honokiol in preparing natural environment-friendly pesticide.

2. Use according to claim 1, characterized in that: is used for preventing and treating crop bacterial diseases and fungal diseases.

3. Use according to claim 2, characterized in that: the bacterial disease is bacterial wilt.

4. Use according to claim 2, characterized in that: the fungal diseases are tobacco phytophthora parasitica, tobacco brown spot, tomato wilt, cucumber anthracnose, wheat sharp eyespot, apple alternaria leaf spot, tomato gray mold or grape gray mold.

5. Use according to claim 2, characterized in that: when the honokiol is used for preventing and treating crop bacterial diseases, the final concentrations of the honokiol are respectively 80 mug/mL, 64 mug/mL, 40 mug/mL, 32 mug/mL, 20 mug/mL, 16 mug/mL and 10 mug/mL.

6. Use according to claim 2, characterized in that: when the honokiol is used for preventing and treating fungal diseases of crops, the final concentrations of the honokiol are respectively 64 mu g/mL, 40 mu g/mL, 32 mu g/mL, 20 mu g/mL, 16 mu g/mL, 10 mu g/mL and 8 mu g/mL.

7. Use according to claim 2, characterized in that: when the composition is used for preventing and treating fungal diseases of crops, the composition is combined with magnolol and sodium remains ampicillin.

Technical Field

The invention relates to application of honokiol in preparation of natural environment-friendly pesticides, and belongs to the technical field of new application of honokiol.

Background

The control of the crop bacterial wilt is a worldwide problem, called plant cancer, and particularly after agricultural streptomycin is stopped in China, efficient and environment-friendly alternative medicaments are urgently needed in production. The existing prevention and treatment agents are mainly copper agents, have poor prevention and treatment effects and pollute the environment.

The prevention and treatment of fungal diseases such as tobacco black shank, tomato and the like are mainly chemical agents such as metalaxyl, dimethomorph and the like, and are easy to cause negative effects such as pesticide residue, environmental pollution, pathogenic bacteria drug resistance and the like. In addition, the tomato gray mold and the grape gray mold are worldwide crop diseases caused by botrytis cinerea infection, are higher in incidence in fruits and vegetables such as tomatoes, grapes and the like, and cause the problems of food pollution, environmental pollution and the like caused by long-term use of chemical pesticides.

Therefore, it is required to find a botanical pesticide which is derived from plants, safe to people and livestock, capable of maintaining the quality of agricultural products and playing a role in preventing fungal diseases of crops.

CN201910475882.X discloses an application of honokiol in fruit and vegetable preservation and a preparation prepared from honokiol for controlling postharvest botrytis of fruits and vegetables, mainly relates to the application in fruit and vegetable preservation, and only discovers the effect of honokiol on controlling postharvest botrytis.

Disclosure of Invention

The invention aims to provide the application of honokiol in preparing natural environment-friendly pesticides, provides a new safe and effective substance for preventing and treating crop diseases, improves the prevention and treatment effect and reduces the pesticide application cost.

The invention provides an application of honokiol in preparation of natural environment-friendly pesticides.

The molecular weight of the honokiol is 266.33, and the molecular formula is C₁₈H₁₈0₂。

The application is used for preventing and treating crop bacterial diseases and fungal diseases.

The bacterial disease is bacterial wilt.

The fungal diseases are tobacco phytophthora parasitica, tobacco brown spot, tomato wilt, cucumber anthracnose, wheat sharp eyespot, apple alternaria leaf spot, tomato gray mold or grape gray mold.

When the honokiol is used for preventing and treating crop bacterial diseases, the final concentrations of the honokiol are respectively 80 mug/mL, 64 mug/mL, 40 mug/mL, 32 mug/mL, 20 mug/mL, 16 mug/mL and 10 mug/mL.

When the honokiol is used for preventing and treating fungal diseases of crops, the final concentrations of the honokiol are respectively 64 mu g/mL, 40 mu g/mL, 32 mu g/mL, 20 mu g/mL, 16 mu g/mL, 10 mu g/mL and 8 mu g/mL.

When the honokiol is used for preventing and treating fungal diseases, cell walls of hyphae are mainly damaged by the honokiol, leakage of hyphae contents is caused in 2 hours, the sterilization time is short, and the effect is quick.

The invention discovers the bacteriostatic effect of honokiol on 1 plant bacterial disease and 8 plant fungal diseases, and is mainly applied before the plants grow and are picked in the field to control the occurrence of plant diseases from the source. In addition, the invention defines the optimal bactericidal concentration of honokiol when the plant diseases are not used, ensures the control of the diseases and simultaneously keeps the lowest agricultural cost.

Compared with the prior art, the invention has the following beneficial effects:

the botanical pesticide is a pesticide agent processed by utilizing the components of plant roots, stems, leaves and the like, is derived from plants, is safe to people and livestock, can maintain the quality of agricultural products, and can play a role in preventing fungal diseases of crops. Therefore, compared with the existing chemical pesticide, the botanical fungicide provided by the invention is separated from magnolia officinalis, can prevent and treat the above 9 plant diseases, is environment-friendly, harmless to human and livestock, strong in production pertinence, capable of improving the prevention and treatment effect, reducing the pesticide application cost and ensuring the safety of fruits and grains.

Drawings

FIG. 1 is a graph showing the bacteriostatic effect of honokiol on alternaria alternate and tobacco black shank;

FIG. 2 is a graph showing the bacteriostatic effects of honokiol on tomato wilt, tomato gray mold, cucumber anthracnose, apple spot, grape gray mold, and wheat sharp eyespot;

FIG. 3 is a graph showing the relationship between the time of destruction and the concentration of honokiol on hyphae of a fungal disease (expressed by conductivity).

Detailed Description

The present invention will be described in detail below with reference to specific examples, but the present invention is not limited to these examples.

Examples

Materials and methods

1. Test strains

Pathogenic bacteria of tobacco bacterial wilt (Ralstonia solanacearum), tobacco black shank (Phytophthora nicotianae) and tobacco brown spot (friends) are obtained by separating and purifying infected tobacco strains in a tobacco pest monitoring and comprehensive treatment key laboratory of tobacco institute of Chinese academy of agricultural sciences;

tomato blight (Fusarium oxysporum f.sp.lycopersico), Cucumber anthracnose (Cucumber anthracnose), wheat sharp eyespot (Rhizoctonia cerealis), apple Alternaria leaf spot (Alternaria mali rob.), tomato gray mold (Botrytis cinerea per. ex Fr.) and grape gray mold (Botrytis cinerea) are provided by the chemical and pharmaceutical academy of the Qingdao university of agriculture.

2. Honokiol activity in inhibiting bacterial wilt

Inhibit medium concentrationMeasurement of degree (EC)₅₀). The bacterial wilt inhibitory activity of honokiol is measured on a 96-hole cell culture plate. Washing bacterial wilt activated for 12h on NA slant culture medium with sterile water, and diluting to bacterial content of about 10⁸And (3) adding honokiol into CFU/mL, adding honokiol into culture wells with final concentrations of 80 μ g/mL, 64 μ g/mL, 40 μ g/mL, 32 μ g/mL, 20 μ g/mL, 16 μ g/mL and 10 μ g/mL respectively, adding 200 μ L of sterile water as a control, shaking the bacteria for 24h under the conditions of 28 ℃ and 180r/min, and calculating EC50 by setting three times for each group.

3. Honokiol inhibiting mycosis activity

A growth rate method is used. The honokiol is prepared into OA plates with different concentrations by DMSO (0.5%, v/v), the final concentration of the honokiol is 64 mu g/mL, 40 mu g/mL, 32 mu g/mL, 20 mu g/mL, 16 mu g/mL, 10 mu g/mL and 8 mu g/mL, 100 mu L of ampicillin sodium is added, the solidification is carried out, the fungus inoculation cake is 0.5cm, 3 repeated experiments are set for each concentration, and the culture is carried out for 7 days at the temperature of 28 ℃. DMSO with the same dilution ratio is used as control, the colony diameter is measured by cross method, and the inhibition rate of hypha growth and EC are calculated₅₀The value is obtained.

(II) results and analysis

1. Honokiol activity for inhibiting tobacco bacterial wilt

MIC determination results show that honokiol is higher than 64 mu g.mL^-1Under the concentration treatment, no bacterial wilt growth is seen, which indicates that the MIC of honokiol to bacterial wilt is 64 mu g.mL^-1。

As shown in Table 1, honokiol has good inhibitory effect on Ralstonia solanacearum, and the toxicity equation is Y-3.0176 x +1.8253, R²0.9864, EC50 value 11.27/. mu.g.mL^-1。

TABLE 1 determination of inhibitory Effect of Honokiol on tobacco bacterial wilt

Honokiol dosage (μ L/L)	Bacteriostatic ratio (%)
		0	-
10	47.07
		16	61.86
20	64.76
		32	82.16
40	96.26
		64	98.68

2. Honokiol inhibiting activity of various crop fungus diseases

As shown in table 2, honokiol has different bacteriostatic effects on different fungal diseases of crops.

The honokiol has good bacteriostatic effect on tobacco black shank, and the regression equation of toxicity is Y-2.6448X +1.2166 (R)²＝0.9916)，EC₅₀26.95. mu.g/mL^-1；

The honokiol has good bacteriostatic effect on the alternaria alternate, and the toxicity regression equation is Y-1.7280X +2.4006 (R)²＝0.9890)，EC₅₀31.94. mu.g.mL^-1；

The honokiol has good bacteriostatic effect on tomato wilt, and the toxicity regression equation is Y-1.8778X +2.5205(R is²＝0.9745)，EC₅₀20.91. mu.g/mL^-1；

The honokiol has good bacteriostatic effect on cucumber anthracnose, and the regression equation of virulence is Y-1.4594X +3.0704 (R)²＝0.9953)，EC₅₀21.00. mu.g/mL^-1；

The honokiol has strong bacteriostatic effect on wheat sharp eyespot, and the toxicity regression equation is Y-2.5376X +1.9671 (R)²＝0.9884)，EC₅₀15.67. mu.g/mL^-1；

The honokiol has good bacteriostatic effect on alternaria mali, and the toxicity regression equation is Y-0.9297X +3.7619 (R)²＝0.9904)，EC₅₀21.46. mu.g/mL^-1；

The honokiol has good bacteriostatic effect on tomato gray mold, and the regression equation of toxicity is that Y is 0.7402X +3.8678(R is²＝0.9955)，EC₅₀33.85. mu.g/mL^-1；

The honokiol has strong bacteriostatic effect on botrytis cinerea drug resistance, and the toxicity regression equation is that Y is 0.4518X +4.7511(R is²＝0.9850)，EC₅₀3.56. mu.g/mL^-1。

TABLE 2-1 determination of the fungicidal Effect of honokiol on crop diseases

TABLE 2-2 measurement of the bacteriostatic effect of honokiol on fungal diseases of crops