CN114176095A - Bacteriostatic liquid for preventing and treating fungal plant diseases and preparation method and use method thereof - Google Patents

Bacteriostatic liquid for preventing and treating fungal plant diseases and preparation method and use method thereof Download PDF

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CN114176095A
CN114176095A CN202111603879.5A CN202111603879A CN114176095A CN 114176095 A CN114176095 A CN 114176095A CN 202111603879 A CN202111603879 A CN 202111603879A CN 114176095 A CN114176095 A CN 114176095A
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water
solution
bacteriostatic
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silicon element
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王忠厚
李克明
杨宗兰
李奎江
经文静
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Huaxia Ruitian Beijing Biotechnology Co ltd
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N59/00Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
    • A01N59/02Sulfur; Selenium; Tellurium; Compounds thereof
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N59/00Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08Magnoliopsida [dicotyledons]
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08Magnoliopsida [dicotyledons]
    • A01N65/20Fabaceae or Leguminosae [Pea or Legume family], e.g. pea, lentil, soybean, clover, acacia, honey locust, derris or millettia
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08Magnoliopsida [dicotyledons]
    • A01N65/36Rutaceae [Rue family], e.g. lime, orange, lemon, corktree or pricklyash
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/40Liliopsida [monocotyledons]
    • A01N65/42Aloeaceae [Aloe family] or Liliaceae [Lily family], e.g. aloe, veratrum, onion, garlic or chives

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  • General Health & Medical Sciences (AREA)
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  • Natural Medicines & Medicinal Plants (AREA)
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  • Wood Science & Technology (AREA)
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  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The invention relates to the technical field of plant disease prevention, in particular to a bacteriostatic liquid for preventing and treating fungal diseases of plants, a preparation method and a use method thereof. The invention provides a bacteriostatic liquid for preventing and treating fungal diseases of plants, which is prepared from the following raw materials in parts by mass: 10-20 parts of water-soluble silicon element, 25-35 parts of traditional Chinese medicine composite extracting solution, 3-7 parts of sulfur and 45-65 parts of water. The bacteriostatic liquid contains the water-soluble silicon element and the compound extracting solution of the Chinese herbal medicines, and can improve the drug effect in the extraction of the active ingredients of the Chinese herbal medicines by utilizing the ultra-strong permeability of the water-soluble silicon element which can completely dissolve the sulfur and the water-soluble silicon element, so that the bacteriostatic liquid has quick effect, obvious effect and stronger bacteriostatic action.

Description

Bacteriostatic liquid for preventing and treating fungal plant diseases and preparation method and use method thereof
Technical Field
The invention relates to the technical field of plant disease prevention, in particular to a bacteriostatic liquid for preventing and treating fungal diseases of plants, a preparation method and a use method thereof.
Background
Among plant diseases, 70-80% of the diseases are caused by pathogenic fungi infection. The plant fungal diseases refer to diseases that plant roots, stems or leaves are infected by fungal pathogenic bacteria living in soil or air under proper conditions, so that plants are wilted and died, even fruits are damaged, so that the fruits are diseased or rotted, and the yield is influenced. The plant fungal diseases not only directly cause the reduction of the yield and the quality of crops, but also cause a plurality of toxins and metabolites harmful to human and livestock to be secreted and generated by part of pathogenic fungi in the process of infecting the crops, thereby forming a great threat to the safety of agricultural products. The fungal pathogens are spread in various ways, infect and cause systemic diseases of plants, and cause great loss to agricultural production, and common fungal diseases of crops mainly comprise anthracnose, rust diseases, powdery mildew, downy mildew, leaf blight, soot disease and the like. At present, the types and the quantity of medicaments for preventing and treating the fungal diseases in China are few and few, and the medicaments are mostly copper preparations and antibiotic medicaments, and the new varieties are few. After long-term use, pesticide residue, environmental pollution and drug resistance of germs are increasingly serious, and the prevention and treatment effect of the existing medicament is increasingly reduced.
At present, the control of plant fungal diseases is mostly chemical control and microbial control. Chinese patent document CN102972398A discloses an application of a compound in controlling plant fungal diseases caused by alternaria alternata, in particular to an application of 2-methyl-5-isopropyl phenol in controlling plant fungal diseases caused by alternaria alternata, and the invention content is as follows: the OP-10 emulsifier is added into the 2-methyl-5-isopropyl phenol according to the proportion of 1-45% of the total volume and dissolved by acetone to prepare a 1-80% (w/w) 2-methyl-5-isopropyl phenol solution which is used for preventing and treating tomato early blight, cabbage black spot and tobacco brown spot caused by alternaria alternate. However, the chemical bactericide not only pollutes the environment in production and has high production cost, but also inevitably causes environmental pollution and agricultural product pesticide residue problems by repeated application.
Therefore, the development of novel, highly effective, low-toxicity, environment-friendly fungal disease control agents is very necessary and urgent.
Disclosure of Invention
The invention aims to provide the bacteriostatic liquid for preventing and treating the fungal diseases of plants, which contains the water-soluble silicon element and the compound extracting solution of the Chinese herbal medicines, and can increase the drug effect in the extraction of the active ingredients of the Chinese herbal medicines by utilizing the ultra-strong permeability of the water-soluble silicon element which can completely dissolve the sulfur and the water-soluble silicon element, so the bacteriostatic liquid has quick response, obvious effect and stronger bacteriostatic action.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a bacteriostatic liquid for preventing and treating fungal diseases of plants, which is prepared from the following raw materials in parts by mass: 10-20 parts of water-soluble silicon element, 25-35 parts of traditional Chinese medicine composite extracting solution, 3-7 parts of sulfur and 45-65 parts of water.
Preferably, the traditional Chinese medicine compound extracting solution is obtained by mixing and extracting raw materials including radix sophorae flavescentis, rhizoma smilacis glabrae, pokeberry root, ephedra, lithospermum and golden cypress; the mass ratio of the radix sophorae flavescentis to the rhizoma smilacis glabrae to the radix phytolaccae is 15-25: 15-25: 15-25: 15-25: 8-12: 8 to 12.
Preferably, the extracted extracting solution is a water solution of water-soluble silicon element, and the mass percentage of the water-soluble silicon element in the water solution of the water-soluble silicon element is 0.5-1.5%; the extraction method comprises decocting.
Preferably, the mass ratio of the raw materials to the extracting solution is 1.5-2.5: 90-110.
Preferably, the decocting time is 30-50 min.
Preferably, the mass concentration of the water-soluble silicon element is 8-12%.
The invention also provides a preparation method of the antibacterial liquid for preventing and treating fungal diseases of plants, which comprises the following steps:
(1) mixing water-soluble silicon element with sulfur, and decocting to obtain solution 1;
(2) mixing the solution 1 obtained in the step (1), the traditional Chinese medicine compound extracting solution and water.
Preferably, the decocting time in the step (1) is 1.5-2.5 h.
The invention further provides the bacteriostatic solution for preventing and treating the plant fungal diseases, and an application method of the bacteriostatic solution for preventing and treating the plant fungal diseases, which is prepared by the preparation method of the bacteriostatic solution for preventing and treating the plant fungal diseases.
Preferably, the dilution multiple is 150-250 times;
preferably, the plant is a fruit tree;
preferably, each fruit tree is sprayed with 100-200 ml of the diluted antibacterial solution before dilution.
Preferably, the fruit tree is an apple tree.
Compared with the prior art, the invention has the following excellent effects:
1. the bacteriostatic liquid contains the water-soluble silicon element and the compound extracting solution of the Chinese herbal medicines, and can improve the drug effect in the extraction of the active ingredients of the Chinese herbal medicines by utilizing the ultra-strong permeability of the water-soluble silicon element which can completely dissolve the sulfur and the water-soluble silicon element, so that the bacteriostatic liquid has quick effect, obvious effect and stronger bacteriostatic action.
2. The added sophora flavescens, rhizoma smilacis glabrae, pokeberry root, ephedra herb, lithospermum and phellodendron have sterilization effects, and the sulfur also has super strong sterilization. The water-soluble silicon element and the sulfur are mixed and decocted, and the sterilization effect is doubled with little effort. The water-soluble silicon element also has super-strong permeability, and active ingredients in the Chinese herbal medicines can be effectively extracted by decocting the water, the water-soluble silicon element and the Chinese herbal medicines together, so that the sterilization effect of the antibacterial liquid is improved.
3. The bacteriostatic solution has an obvious effect when being sprayed on the leaf surfaces once, and the crops can recover to grow normally after being sprayed twice.
Detailed Description
The invention provides a bacteriostatic liquid for preventing and treating fungal diseases of plants, which is prepared from the following raw materials in parts by mass: 10-20 parts of water-soluble silicon element, 25-35 parts of traditional Chinese medicine composite extracting solution, 3-7 parts of sulfur and 45-65 parts of water;
the compound is preferably prepared from the following raw materials in parts by mass: 12-18 parts of water-soluble silicon element, 27-33 parts of traditional Chinese medicine composite extracting solution, 3-6 parts of sulfur and 49-61 parts of water;
the composite material is further preferably prepared from the following raw materials in parts by mass: 14-16 parts of water-soluble silicon element, 29-31 parts of traditional Chinese medicine composite extracting solution, 4-5 parts of sulfur and 53-57 parts of water;
more preferably, the compound is prepared from the following raw materials in parts by mass: 15 parts of water-soluble silicon element, 30 parts of traditional Chinese medicine compound extracting solution, 4.5 parts of sulfur and 55 parts of water.
In the invention, the water-soluble silicon element is preferably a solution system taking water as a solvent, and the mass concentration of the water-soluble silicon element is 8-12%; preferably 9-11%; further preferably 10%; in the specific implementation process of the invention, the water-soluble silicon element is purchased from Beijing Huaxia rui Tian Biotech Co.
In the invention, the traditional Chinese medicine compound extracting solution is obtained by mixing and extracting raw materials including radix sophorae flavescentis, rhizoma smilacis glabrae, pokeberry root, ephedra herb, lithospermum and golden cypress; the mass ratio of the radix sophorae flavescentis to the rhizoma smilacis glabrae to the radix phytolaccae is 15-25: 15-25: 15-25: 15-25: 8-12: 8 to 12.
Preferably 17-23: 17-23: 17-23: 17-23: 9-11: 9-11;
more preferably 19 to 21: 19-21: 19-21: 19-21: 10: 10;
more preferably 20: 20: 20: 20: 10: 10.
in the invention, the extracted extracting solution is preferably an aqueous solution of water-soluble silicon element, and the mass percentage of the water-soluble silicon element in the aqueous solution of the water-soluble silicon element is 0.5-1.5%, preferably 1%; the extraction method comprises the steps of decocting;
in the invention, the mass ratio of the raw materials to the extracting solution is 1.5-2.5: 90-110; preferably 1.7-2.3: 94-106; further preferably 1.9-2.1: 98-102; more preferably 2: 100; the decocting time is 30-50 min; preferably 34-46 min; further preferably 38-42 min; more preferably 40 min. The invention carries out filtration after the decoction, and the filtration is carried out by adopting a filter or a filtration method recorded in the prior art.
The invention also provides a preparation method of the antibacterial liquid for preventing and treating fungal diseases of plants, which comprises the following steps:
(1) mixing water-soluble silicon element with sulfur, and decocting to obtain solution 1;
(2) mixing the solution 1 obtained in the step (1), the traditional Chinese medicine compound extracting solution and water.
In the invention, the decocting time in the step (1) is 1.5-2.5 h; preferably for 2 hours.
The invention further provides the bacteriostatic solution for preventing and treating the plant fungal diseases and the use method of the bacteriostatic solution for preventing and treating the plant fungal diseases, which is prepared by the preparation method of the bacteriostatic solution for preventing and treating the plant fungal diseases.
In the invention, the dilution multiple is 150-250 times; preferably 170-230 times; further preferably 190-210 times; more preferably 200 times.
In the present invention, the plant is preferably a fruit tree, and the fruit tree is preferably an apple tree.
In the invention, 100-200 ml of the diluted antibacterial solution is sprayed on each fruit tree; preferably 120-180 ml; further preferably 140 to 160 ml; more preferably 150 ml.
The technical solutions provided by the present invention 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
A bacteriostatic liquid for preventing and treating fungal plant diseases is prepared from the following raw materials in parts by weight: 10 parts of water-soluble silicon element, 25 parts of traditional Chinese medicine compound extracting solution, 3 parts of sulfur and 45 parts of water;
the mass concentration of the water-soluble silicon element is 12%;
the traditional Chinese medicine compound extracting solution is obtained by mixing and extracting raw materials including radix sophorae flavescentis, rhizoma smilacis glabrae, pokeberry root, ephedra herb, lithospermum and golden cypress; the mass ratio of the radix sophorae flavescentis to the rhizoma smilacis glabrae to the radix phytolaccae is 15: 15: 15: 15: 8: 8;
the extracted extracting solution is a water solution of water-soluble silicon element, and the mass percentage of the water-soluble silicon element in the water solution of the water-soluble silicon element is 1%; the extraction method comprises the steps of decocting;
mixing the raw materials with the extractive solution at a mass ratio of 2.5: 110, decocting for 50min, and filtering to obtain Chinese medicinal composite extractive solution;
the preparation method of the bacteriostatic liquid for preventing and treating the fungal diseases of plants comprises the following steps:
(1) mixing water-soluble silicon element with sulfur, and decocting for 2.5h to obtain solution 1;
(2) mixing the solution 1 obtained in the step (1), the traditional Chinese medicine compound extracting solution and water.
Example 2
A bacteriostatic liquid for preventing and treating fungal plant diseases is prepared from the following raw materials in parts by weight: 20 parts of water-soluble silicon element, 35 parts of traditional Chinese medicine composite extracting solution, 7 parts of sulfur and 65 parts of water;
the mass concentration of the water-soluble silicon element is 12%;
the traditional Chinese medicine compound extracting solution is obtained by mixing and extracting raw materials including radix sophorae flavescentis, rhizoma smilacis glabrae, pokeberry root, ephedra herb, lithospermum and golden cypress; the mass ratio of the radix sophorae flavescentis to the rhizoma smilacis glabrae to the radix phytolaccae is 25: 25: 25: 25: 12: 12;
the extracted extracting solution is a water solution of water-soluble silicon element, and the mass percentage of the water-soluble silicon element in the water solution of the water-soluble silicon element is 1.5%; the extraction method comprises the steps of decocting;
mixing the raw materials with the extractive solution at a mass ratio of 2.5: 110, decocting for 50min, and filtering to obtain Chinese medicinal composite extractive solution;
the preparation method of the bacteriostatic liquid for preventing and treating the fungal diseases of plants comprises the following steps:
(1) mixing water-soluble silicon element with sulfur, and decocting for 2.5h to obtain solution 1;
(2) mixing the solution 1 obtained in the step (1), the traditional Chinese medicine compound extracting solution and water.
Example 3
A bacteriostatic liquid for preventing and treating fungal plant diseases is prepared from the following raw materials in parts by weight: 15 parts of water-soluble silicon element, 30 parts of traditional Chinese medicine compound extracting solution, 5 parts of sulfur and 55 parts of water;
the mass concentration of the water-soluble silicon element is 10 percent;
the traditional Chinese medicine compound extracting solution is obtained by mixing and extracting raw materials including radix sophorae flavescentis, rhizoma smilacis glabrae, pokeberry root, ephedra herb, lithospermum and golden cypress; the mass ratio of the radix sophorae flavescentis to the rhizoma smilacis glabrae to the radix phytolaccae is 20: 20: 20: 20: 10: 10;
the extracted extracting solution is a water solution of water-soluble silicon element, and the mass percentage of the water-soluble silicon element in the water solution of the water-soluble silicon element is 1%; the extraction method comprises the steps of decocting;
mixing the raw materials with the extractive solution at a mass ratio of 2: 100, decocting for 40min, and filtering to obtain Chinese medicinal composite extractive solution;
the preparation method of the bacteriostatic liquid for preventing and treating the fungal diseases of plants comprises the following steps:
(1) mixing water-soluble silicon element with sulfur, and decocting for 2h to obtain solution 1;
(2) mixing the solution 1 obtained in the step (1), the traditional Chinese medicine compound extracting solution and water.
Example 4
Test subjects: diseased apple trees with brown spot (brown spot of apple is caused by fungal disease, mainly harming apple leaves).
The test method comprises the following steps: the bacteriostat for controlling plant fungal diseases obtained in the example 3 is diluted according to the proportion of 1: 100, 1: 150, 1: 200, 1: 250 and 1: 300 respectively, and a clear water group is used as a blank control group. The diseased apple trees with brown spot are observed and detected by spraying 3 diseased apple trees in each treatment, the percentage of the leaves with diseased spots of each apple tree before spraying, 10 days after spraying for the first time and 10 days after spraying for the second time is respectively counted, the average value is taken, and the result is shown in table 1.
The spraying method comprises the following steps: spraying 150ml of diluted antibacterial solution to each fruit tree, and continuously spraying for 2 times at intervals of 10 days.
TABLE 1 statistical results
Figure BDA0003432938480000061
Figure BDA0003432938480000071
As is apparent from the description in Table 1, the bacteriostatic agent for controlling fungal plant diseases obtained in example 3 was diluted at a ratio of 1: 200, and had an excellent effect of controlling cercospora leaf spot of apple. The 1: 150 and 1: 250 treatment groups had slightly poor control of apple brown spot. As can be seen from the treatment groups of 1: 100 and 1: 300, the dilution ratio of the bacteriostatic solution for preventing and treating the fungal diseases of plants cannot be too large or too small, otherwise, the technical effect of effectively preventing and treating the cercospora leaf spot of the apple cannot be achieved.
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 (10)

1. The antibacterial liquid for preventing and treating fungal plant diseases is characterized by being prepared from the following raw materials in parts by mass: 10-20 parts of water-soluble silicon element, 25-35 parts of traditional Chinese medicine composite extracting solution, 3-7 parts of sulfur and 45-65 parts of water.
2. The bacteriostatic liquid for preventing and treating the fungal diseases of plants according to claim 1, wherein the traditional Chinese medicine compound extracting solution is obtained by mixing and extracting raw materials comprising radix sophorae flavescentis, rhizoma smilacis glabrae, pokeberry root, ephedra herb, lithospermum and golden cypress; the mass ratio of the radix sophorae flavescentis to the rhizoma smilacis glabrae to the radix phytolaccae is 15-25: 15-25: 15-25: 15-25: 8-12: 8 to 12.
3. The bacteriostatic liquid for controlling the fungal plant diseases according to claim 2, wherein the extracted extracting solution is an aqueous solution of water-soluble silicon element, and the mass percentage of the water-soluble silicon element in the aqueous solution of water-soluble silicon element is 0.5-1.5%; the extraction method comprises decocting.
4. The bacteriostatic liquid for controlling the fungal diseases of plants according to claim 3, wherein the mass ratio of the raw materials to the extracting solution is 1.5-2.5: 90-110.
5. The bacteriostatic solution for controlling plant fungal diseases according to claim 3 or 4, wherein the decocting time is 30-50 min.
6. The bacteriostatic solution for controlling plant fungal diseases according to claim 1, wherein the mass concentration of the water-soluble silicon element is 8-12%.
7. The preparation method of the bacteriostatic liquid for controlling the fungal diseases of plants as claimed in any one of claims 1 to 6, characterized by comprising the following steps:
(1) mixing water-soluble silicon element with sulfur, and decocting to obtain solution 1;
(2) mixing the solution 1 obtained in the step (1), the traditional Chinese medicine compound extracting solution and water.
8. The method for preparing the bacteriostatic liquid for controlling the fungal diseases of plants according to claim 7, wherein the decocting time in the step (1) is 1.5-2.5 h.
9. The use of the bacteriostatic solution for controlling plant fungal diseases according to any one of claims 1 to 6 or the bacteriostatic solution for controlling plant fungal diseases prepared by the preparation method according to claim 7 or 8, wherein the bacteriostatic solution for controlling plant fungal diseases is diluted and sprayed on plants.
10. The use of claim 9, wherein the dilution factor is 150-250.
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