CN111990415A

CN111990415A - Botanical fungicide and preparation method and application thereof

Info

Publication number: CN111990415A
Application number: CN202010792107.XA
Authority: CN
Inventors: 曹挥; 谷晓杰; 池吉平; 李少华; 冯雪
Original assignee: Shanxi Agricultural University; Thai Nguyen University
Current assignee: Shanxi Agricultural University; Thai Nguyen University
Priority date: 2020-08-08
Filing date: 2020-08-08
Publication date: 2020-11-27

Abstract

The invention provides a botanical fungicide, a preparation method and application thereof, relating to the technical field of novel fungicides; the bactericide takes the extracts of the star anise, the grassleaf sweelflag rhizome and the macleaya cordata as effective components, is natural in material taking, is favorable for saving cost, belongs to a pure Chinese medicinal preparation, is easy to decompose under natural conditions, cannot pollute soil and underground water, and cannot cause harm to animals and plants; the extraction of active ingredients in the plant is simple and easy, and is beneficial to industrial production; is not easy to generate drug resistance. Therefore, the bacteriostatic agent has a strong inhibiting effect on apple ring spot and can fundamentally prevent and treat apple ring spot.

Description

Botanical fungicide and preparation method and application thereof

Technical Field

The invention belongs to the technical field of novel bactericides, and particularly relates to a botanical bactericide as well as a preparation method and application thereof.

Background

Apple ring rot, also known as verruca or rough skin disease, is a fungal disease caused by plasmodiophora beijerinckii, which mainly compromises the branches, tender tips and fruits of apple trees. The disease occurs in the major apple producing areas of China, Japan, and Korea. The disease is more serious in the middle of China in high-temperature and rainy areas, the perennial rotten fruit rate can reach 20-30%, and the rainy year can reach 40-50% or even 60-70%. Therefore, the ring rot of apple becomes one of the most serious diseases in apple production in China at present, and simultaneously causes great economic loss to the apple industry in China.

At present, the prevention and the treatment of the apple ring spot mainly depend on chemical pesticides, such as tebuconazole, carbendazim and mancozeb. The long-term use of chemical pesticides can cause diseases to generate drug resistance and cause pesticide residues to harm human health.

Disclosure of Invention

In view of the above, the invention aims to provide a botanical fungicide and a preparation method and application thereof, the material is natural and pollution-free, no pesticide residue is generated, the plant bacteriostatic component has strong inhibiting effect on apple ring rot, and the apple ring rot can be fundamentally prevented and treated.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a botanical fungicide, which comprises the following raw materials in percentage by mass: 18-25% of star anise extract, 3-8% of grassleaf sweelflag rhizome extract, 3-8% of macleaya cordata extract, 8-12% of solubilizer, 8-12% of emulsifier, 0.8-1.2% of thickener, 0.8-1.2% of antifreezing agent and the balance of water.

Preferably, the extraction solvent of the star anise extract, the grassleaf sweelflag rhizome extract and the macleaya cordata extract is one or more of methanol, ethanol and acetone.

Preferably, the volume ratio of the star anise extract, the grassleaf sweelflag rhizome extract and the macleaya cordata extract to the extraction solvent is 1: 5.

Preferably, the preparation methods of the fennel extract, the grassleaf sweelflag rhizome extract and the macleaya cordata extract are as follows: respectively cleaning fructus Anisi Stellati, rhizoma Acori Graminei and herba Macleayae Cordatae, oven drying, and pulverizing; respectively mixing the powder and an extraction solvent according to the volume ratio of 1:5, leaching for 12 hours or continuously refluxing for 4-8 hours, filtering, and concentrating under reduced pressure to obtain an extract stock solution.

Preferably, the emulsifier comprises one or more of emulsifier 1601, dispersant 123K, emulsifier EL-40, and tween 80.

Preferably, the thickening agent comprises one or more of xanthan gum, gum arabic, polyvinylpyrrolidone.

Preferably, the antifreeze agent comprises one or more of ethylene glycol, glycerol and isopropanol.

The invention also provides a preparation method of the bactericide, which comprises the following steps: mixing the star anise extract, the grassleaf sweelflag rhizome extract, the macleaya cordata extract, the solubilizer, the emulsifier, the thickener, the antifreezing agent and water according to the proportion to obtain the bactericide.

The invention also provides application of the bactericide in inhibiting and/or treating apple ring spot.

Preferably, Ec of the fungicide₅₀The value was 0.2650 mg/ml.

The invention provides a botanical fungicide, which takes extracts of star anise, grassleaf sweelflag rhizome and macleaya cordata as effective components, is natural in material selection, is favorable for saving cost, belongs to a pure traditional Chinese medicine preparation, is easy to decompose under natural conditions, does not pollute soil and underground water, and does not cause harm to animals and plants; the extraction of active ingredients in the plant is simple and easy, and is beneficial to industrial production; all the plant extracts are processed by plant extracts, and the plant extracts are a multi-component mixture and have a plurality of action sites for pathogenic bacteria, so that the plant extracts are not easy to generate drug resistance. In the embodiment of the invention, the bacteriostasis effect of the bactericide is verified through a bacteriostasis zone experiment, and finally the regression equation of the toxicity is determined to be 7.0710+3.5906 and Ec₅₀The value is 0.2650mg/ml, the correlation coefficient is 0.7600, and the bacteriostatic agent has stronger inhibition effect on the physalospora piricolaThe preparation method can fundamentally prevent and treat the apple ring spot.

Drawings

FIG. 1 is a graph showing the inhibition of mycelial growth of apple ring rot by the fungicide of the present invention, wherein A is a control group and B is an experimental treatment group.

Detailed Description

The bactericide of the invention preferably comprises the following raw materials in percentage by mass: 19-22% of star anise extract, 4-6% of grassleaf sweelflag rhizome extract, 4-6% of macleaya cordata extract, 9-10% of solubilizer, 9-10% of emulsifier, 0.9-1% of thickener, 1% of antifreeze and the balance of water, and more preferably comprises 20% of star anise extract, 5% of grassleaf sweelflag rhizome extract, 5% of macleaya cordata extract, 10% of solubilizer, 10% of emulsifier, 1% of thickener, 1% of antifreeze and the balance of water.

The extraction solvent of the star anise extract, the grassleaf sweelflag rhizome extract and the macleaya cordata extract is preferably one or more of methanol, ethanol and acetone, and more preferably ethanol. During extraction of the illicium verum extract, the acorus gramineus soland extract and the macleaya cordata extract, the volume ratio of the illicium verum, the acorus gramineus soland, the macleaya cordata and the extraction solvent is preferably 1: 5.

In the present invention, the preparation methods of the caraway extract, the grassleaf sweelflag rhizome extract and the macleaya cordata extract are preferably all as follows: respectively cleaning fructus Anisi Stellati, rhizoma Acori Graminei and herba Macleayae Cordatae, oven drying, and pulverizing; respectively mixing the powder and an extraction solvent according to the volume ratio of 1:5, leaching for 12 hours or continuously refluxing for 4-8 hours, filtering, and concentrating under reduced pressure to obtain an extract stock solution. The particle size after pulverization according to the present invention is preferably less than 60 mesh. In the extraction process, the leaching is preferably carried out for 4 times. The temperature of the leaching or continuous reflux is preferably 30-45 ℃.

The emulsifier comprises one or more of an emulsifier 1601, a dispersant 123K, an emulsifier EL-40 and Tween 80, and is more preferably the emulsifier 1601. The thickening agent of the present invention preferably comprises one or more of xanthan gum, arabic gum and polyvinylpyrrolidone, more preferably xanthan gum. The antifreeze of the present invention preferably comprises one or more of ethylene glycol, glycerol, and isopropanol, more preferably glycerol.

When the bactericide is prepared, the three plant extract stock solutions are preferably mixed in proportion and then sequentially added with auxiliary materials such as a solubilizer, an emulsifier, a thickener, an antifreezing agent, a solvent and the like to prepare the bactericide.

Ec of the fungicide of the invention₅₀The value was 0.2650 mg/ml.

The plant-derived fungicide provided by the present invention, its preparation method and application are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.

Example 1

Cleaning collected fructus Anisi Stellati, rhizoma Acori Graminei and herba Macleayae Cordatae, drying in the shade in ventilated place, or oven drying at 45 deg.C, pulverizing respectively, and sieving with 60 mesh sieve. Placing the sieved plant powder into a multifunctional extraction tank, adding solvent ethanol for extraction, repeatedly extracting for 4 times at a ratio of 1:5 for 12 hours each time, collecting the extractive solution, and concentrating under reduced pressure to a certain concentration. Adding 30kg of star anise, grassleaf sweelflag rhizome and macleaya cordata extract into 10kg of solubilizer, adding into a mixing kettle, mixing for 1.5h at room temperature, sequentially adding 10kg of emulsifier, 1kg of thickener, 1kg of antifreeze and 48kg of solvent, and uniformly mixing to obtain the botanical fungicide for preventing and treating apple ring rot.

Example 2

Cleaning collected fructus Anisi Stellati, rhizoma Acori Graminei and herba Macleayae Cordatae, drying in the shade in ventilated place, or oven drying at 45 deg.C, pulverizing respectively, and sieving with 60 mesh sieve. Placing the sieved plant powder into a multifunctional extraction tank, adding acetone as solvent for extraction, and concentrating under reduced pressure. Adding 30kg of star anise, grass-leaved sweetflag rhizome and macleaya cordata extract into 15kg of solubilizer, adding into a mixing kettle, mixing for 3h at room temperature, sequentially adding 5kg of emulsifier, 1kg of thickener, 1kg of antifreeze and 48kg of solvent, and uniformly mixing to obtain the botanical fungicide for preventing and treating apple ring rot.

Example 3

Indoor toxicity determination of bactericide on apple ring rot pathogen

1. Reagent for testing

The plant-derived fungicide prepared in example 1.

2. Test germs

Physalospora piricola (Botryosphaeria).

3. Test method

The hypha growth rate method is adopted for determination: there were 2 treatments, i.e. the biocide and blank control. Adding the aqueous emulsion into a molten PDA culture medium, fully shaking and pouring the aqueous emulsion into a sterilized culture dish, preparing bacterial blocks from the edges of bacterial colonies of bacteria cultured for 4-5 days by using a puncher with the diameter of 0.6cm after the culture medium is solidified, inoculating the bacterial blocks onto a culture medium containing medicines by using an inoculating needle, wherein the hypha face is downward, one bacterial block is placed in each dish, each treatment is repeated for 5 times, culturing is carried out in a constant-temperature incubator at 25 ℃, and the diameter of the bacterial colonies is measured by adopting a cross method after 72 hours. Calculating the bacteriostasis rate according to formula I:

processing data by using Excel and DPS software, calculating bacteriostasis rate, toxicity equation and effective inhibition medium concentration EC₅₀The results are shown in tables 1 to 2 and FIG. 1.

TABLE 1 inhibitory effect of this fungicide on apple ring rot pathogen in 48h, 72 h, 96h at different concentrations

TABLE 2 regression equation of virulence, correlation coefficient and concentration in inhibition (EC)₅₀)

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims

1. The botanical fungicide is characterized by comprising the following raw materials in percentage by mass: 18-25% of star anise extract, 3-8% of grassleaf sweelflag rhizome extract, 3-8% of macleaya cordata extract, 8-12% of solubilizer, 8-12% of emulsifier, 0.8-1.2% of thickener, 0.8-1.2% of antifreezing agent and the balance of water.

2. The bactericide as claimed in claim 1, wherein the extraction solvent for the extracts of star anise, grassleaf sweelflag rhizome and macleaya cordata is one or more of methanol, ethanol and acetone.

3. The bactericide as claimed in claim 2, wherein the star anise extract, the grassleaf sweelflag rhizome extract and the macleaya cordata extract are extracted in a volume ratio of 1:5 for each of the star anise, the grassleaf sweelflag rhizome and the macleaya cordata and the extraction solvent.

4. The fungicide according to claim 2 or 3, wherein said caraway extract, grassleaved sweetflag extract and macleaya cordata extract are prepared by the following method: respectively cleaning fructus Anisi Stellati, rhizoma Acori Graminei and herba Macleayae Cordatae, oven drying, and pulverizing; respectively mixing the powder and an extraction solvent according to the volume ratio of 1:5, leaching for 12 hours or continuously refluxing for 4-8 hours, filtering, and concentrating under reduced pressure to obtain an extract stock solution.

5. The bactericide according to claim 1, wherein the emulsifier comprises one or more of emulsifier 1601, dispersant 123K, emulsifier EL-40 and tween 80.

6. The bactericide as claimed in claim 1, wherein said thickener comprises one or more of xanthan gum, arabic gum and polyvinylpyrrolidone.

7. The disinfectant according to claim 1, wherein said antifreezing agent comprises one or more of ethylene glycol, glycerin and isopropyl alcohol.

8. The method for producing the bactericide as claimed in any one of claims 1 to 7, comprising the steps of: mixing the star anise extract, the grassleaf sweelflag rhizome extract, the macleaya cordata extract, the solubilizer, the emulsifier, the thickener, the antifreezing agent and water according to the proportion to obtain the bactericide.

9. Use of the fungicide according to any one of claims 1 to 7 for inhibiting and/or treating ring rot of apple.

10. Use according to claim 9, characterized in that the Ec of the fungicide₅₀The value was 0.2650 mg/ml.