CN110169422B - Compound formulation plant bactericide and preparation method and application thereof - Google Patents
Compound formulation plant bactericide and preparation method and application thereof Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N65/06—Coniferophyta [gymnosperms], e.g. cypress
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- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
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- A01N65/08—Magnoliopsida [dicotyledons]
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- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/22—Lamiaceae or Labiatae [Mint family], e.g. thyme, rosemary, skullcap, selfheal, lavender, perilla, pennyroyal, peppermint or spearmint
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Abstract
The invention relates to a compound-dosage form plant bactericide which is prepared from the following raw materials in percentage by mass: stem and leaf of pseudo-osbeckia, ginkgo leaf, houttuynia cordata, scutellaria baicalensis, isatis root, forsythia, andrographis paniculata, sodium nitrophenolate, OP-10 emulsifier, pseudo-osbeckia flower essential oil, water, wherein the ratio of the water to the pseudo-osbeckia flower essential oil is 20-40: 2-5: 4-16: 2-6: 3-8: 2-8: 1.0-2.5: 0.01-0.03: 0.1-0.25: 10-30. The invention adopts the opacifier which is prepared by taking natural plant resources as the main material and taking the potent cell activator-compound sodium nitrophenolate as the auxiliary material, has the characteristics of environmental protection, no toxicity, high plant absorption and utilization rate and the like, and can be reasonably applied to invasive plants to the utmost extent. The plant source related by the invention is easy to obtain, the preparation process is simpler, the cost is not high, and the method is suitable for agricultural/forestry industrialized production; the plant bactericide has strong killing capability on ralstonia solanacearum and citrus sand-skin disease and also has a certain inhibition effect on pathogenic bacteria of yellow dragon disease.
Description
Technical Field
The invention belongs to the technical field of plant bactericides, and particularly relates to a plant compound bactericide as well as a preparation method and application thereof.
Background
Chinese plant resources are very rich, only the number of medicinal plants reaches 8000, and a considerable part of the medicinal plants contain various bactericidal active ingredients, including terpenes, alkaloids, amides, long-chain carbon alkanes, fatty acids, nitrogen-containing and sulfur-containing compounds, aromatic compounds, phenols, flavonoids, aromatic compounds and the like. The alcohol extract of the pseudo-osmyl grass can reduce the egg laying amount of the diaphorina citri on the new tips of the citrus seedlings, and the essential oil of the alcohol extract has a repellent effect on the adult diaphorina citri; the alcohol extract of herba Houttuyniae has effect in scavenging hydroxyl free radical (. OH), superoxide free radical (O)2 －·) And Nitrite (NO)2 －) The cleaning effect is obvious, and the cleaning agent has the effects of inhibiting and killing various bacteria and partial fungal pathogenic bacteria; the ginkgo leaf contains more than 30 flavone compounds, terpenoids, phenols, trace elements, amino acids and other effective components, and the crude flavone extract has certain bacteriostatic effect on bacteria (staphylococcus aureus and escherichia coli) and fungi (fusarium graminearum and alternaria alternata); the extract of the isatis root has good inhibition and killing effects on bacteria and viruses; fructus forsythiae, common andrographis herb and the like have good inhibition effect on pathogenic bacteria; by utilizing the plant resources with natural bactericidal action and combining with some discovered compounds which are high-efficiency, safe, cheap and easily available and can enhance the stress resistance of plants, the development of novel, green, residue-free and pollution-free environment-friendly bactericides is imperative.
Disclosure of Invention
The invention develops the compound dosage form plant bactericide by integrating the advantages of the plant resources in the aspects of bacteriostasis and sterilization component difference and sterilization effect on different pathogenic microorganisms. The plant bactericide has strong killing capability on ralstonia solanacearum and citrus sand skin disease, and also has a certain inhibition effect on pathogenic bacteria of yellow dragon disease; the invention has no toxicity to human body, the dosage form is emulsion, the use is safe, the plant is easy to absorb after spraying, the invention has no pollution to the environment, and the invention has strong popularization and application value.
In order to solve the technical problems, the invention adopts the technical scheme that: a plant compound bactericide is prepared from the following raw materials in percentage by mass: stem and leaf of pseudo-osbeckia, ginkgo leaf, houttuynia cordata, scutellaria baicalensis, isatis root, forsythia, andrographis paniculata, sodium nitrophenolate, OP-10 emulsifier, pseudo-osbeckia flower essential oil, water, wherein the ratio of the water to the pseudo-osbeckia flower essential oil is 20-40: 2-5: 4-16: 2-6: 3-8: 2-8: 1.0-2.5: 0.01-0.03: 0.1-0.25: 10-30.
In addition, the invention also provides a method for preparing the plant bactericide, which comprises the following steps:
(1) collecting the pseudo-osmyl grass, separately placing the flowers and the stems and leaves, airing the flowers in a shade place, and then placing the pseudo-osmyl grass flowers in an evaporation-condensation container/equipment according to a mass ratio of 1: adding water at a ratio of 20, evaporating at 100 deg.C, and collecting condensate to obtain the essential oil.
(2) Firstly, cutting plant raw materials into pieces, cutting ginkgo leaves, houttuynia cordata and stem leaves of pseudo-rue foetida into strips of 1-2 cm, cutting other wood raw materials into stem sections of about 1cm, uniformly mixing the cut wood raw materials in proportion, pouring the mixture into a reaction kettle, adding water which is 15-20 times of the mass of the plant raw materials, stirring and extracting, wherein the extraction temperature is 80-85 ℃, and the extraction time is 1.5-2.0 hours; and filtering, then performing secondary extraction, controlling the water addition amount to be 8-12 times of the mass of the initial plant raw material, controlling the extraction temperature to be 80-85 ℃, extracting for 1-1.5 hours, filtering, combining the filtrates, heating to 100 ℃, and performing evaporation concentration until the total weight of the plant raw material is 28-35%.
(3) Adding 95% ethanol into the concentrate until the mass percentage of the ethanol is 60% -70%, uniformly stirring, standing for 16-24 h, taking the supernatant to rotary evaporation equipment, evaporating the ethanol to obtain a semi-solidified/viscous substance, and then mixing the semi-solidified/viscous substance according to the mass ratio of 1: adding water in a ratio of 0.8-1.2, mixing uniformly, adding 0.1-0.25 mass percent of pseudo-odorous grass flower essential oil, 1.0-2.5 mass percent of compound sodium nitrophenolate and 0.01-0.03 mass percent of OP-10 emulsifier continuously according to the mass ratio of the plant raw materials, and fully homogenizing by a homogenizer to obtain the plant compound bactericide.
The usage and dosage of the plant bactericide of the invention are as follows: the dosage of the plant protective agent is 0.05 kg-0.3 kg per mu, when the plant protective agent is used, the plant protective agent is firstly diluted by 200-300 times with water, and then the diluted plant bactericide is uniformly sprayed on plants. Has strong killing ability to ralstonia solanacearum and citrus russiamensis, and has certain inhibiting effect to the pathogenic bacteria of yellow dragon disease.
Compared with the prior art, the invention has the following advantages:
1. the plant bactericide of the invention adopts an opacifier which is prepared by taking natural plant resources as a main material and taking the compound sodium nitrophenolate as a powerful cell activator as an auxiliary material, and the invention has the characteristics of environmental protection, no toxicity, high plant absorption and utilization rate and the like.
2. The plant bactericide of the invention takes the invasive plants of China, namely the pseudo-rue foetida as the main raw material, not only provides a new idea for eliminating the invasive plants, but also changes waste into valuable, and rationalizes the application of the invasive plants to the maximum extent.
3. Through detection, the plant bactericide contains the components of volatile essential oil, polysaccharide, flavone, alkaloid and the like, wherein the polysaccharide has the characteristic of protecting immunity, the volatile essential oil and the flavone have the characteristics of inhibiting bacteria and repelling insect pests, and the alkaloid has the function of sterilizing.
4. The plant bactericide has stronger killing capability on ralstonia solanacearum and citrus russiamensis and also has certain inhibition effect on pathogenic bacteria of yellow dragon disease; the invention has no toxicity to human body, the dosage form is emulsion, the use is safe, the plant is easy to absorb after spraying, the invention has no pollution to the environment, and the invention has strong popularization and application value.
Detailed Description
The technical solution of the present invention is further described in detail by the following examples.
Example 1
The plant compound bactericide is prepared from the following raw materials in percentage by mass: stem and leaf of pseudo-osbeckia, ginkgo leaf, houttuynia cordata, scutellaria baicalensis, isatis root, forsythia, andrographis paniculata, sodium nitrophenolate, OP-10 emulsifier, pseudo-osbeckia flower essential oil, water, weight ratio of 30:2:12:4:5:4: 4: 1.5: 0.02: 0.2:25.
The preparation method of the plant protective agent comprises the following steps:
(1) collecting the pseudo-osmyl grass, separately placing the flowers and the stems and leaves, airing the flowers in a shade place, and then placing the pseudo-osmyl grass flowers in an evaporation-condensation container/equipment according to a mass ratio of 1: adding water at a ratio of 20, evaporating at 100 deg.C, and collecting condensate to obtain the essential oil.
(2) Firstly, cutting plant raw materials into pieces, cutting ginkgo leaves, houttuynia cordata and stem leaves of pseudo-odorous grass into strips of 1-2 cm, cutting other wood raw materials into stem sections of about 1cm, uniformly mixing the stem sections according to a proportion, pouring the mixture into a reaction kettle, adding 18 times of water, stirring and extracting, wherein the extraction temperature is 80-85 ℃, and the extraction time is 2.0 hours; and filtering, then carrying out secondary extraction, controlling the water addition amount to be 10 times of the mass of the initial raw materials, controlling the extraction temperature to be 80-85 ℃, extracting for 1.5h, filtering, combining the filtrates, heating to 100 ℃, and carrying out evaporation concentration until the total weight of the plant raw materials is 28-35%.
(3) Adding 95% ethanol into the concentrate until the ethanol content is 70%, uniformly stirring, standing for 24h, taking the supernatant to a rotary evaporation device, evaporating the ethanol to obtain a semi-solidified viscous substance, and mixing the semi-solidified viscous substance with the ethanol according to the mass ratio of 1: adding water according to the proportion of 1, mixing uniformly, adding 0.2 of the pseudo-rue-bane flower essential oil, 1.5 of the compound sodium nitrophenolate and 0.02 of the OP-10 emulsifier according to the mass proportion of the plant raw materials, and fully homogenizing by a homogenizer to obtain the invention.
0.3kg of the plant protective agent is diluted by water by 200 times, and is uniformly sprayed to 1 mu of citrus plants infected with bacterial wilt, so that the bacterial wilt can be effectively prevented and treated after 30 days of application, and the harm of the bacterial wilt of citrus is solved.
Example 2
The plant protective agent is prepared from the following raw materials in percentage by mass: stem and leaf of pseudo-osbeckia, ginkgo leaf, houttuynia, scutellaria, isatis root, forsythia, andrographis, sodium nitrophenolate, OP-10 emulsifier, pseudo-osbeckia flower essential oil, water, weight ratio of 40:3:10:5:4:6:6: 2.0: 0.025: 0.2:20.
The preparation method of the plant protective agent comprises the following steps:
(1) collecting the pseudo-osmyl grass, separately placing the flowers and the stems and leaves, airing the flowers in a shade place, and then placing the pseudo-osmyl grass flowers in an evaporation-condensation container/equipment according to a mass ratio of 1: adding water at a ratio of 20, evaporating at 100 deg.C, and collecting condensate to obtain the essential oil.
(2) Firstly, cutting plant raw materials into pieces, cutting ginkgo leaves, houttuynia cordata and stem leaves of pseudo-odorous grass into strips of 1-2 cm, cutting other wood raw materials into stem sections of about 1cm, uniformly mixing the stem sections according to a proportion, pouring the mixture into a reaction kettle, adding 18 times of water, stirring and extracting, wherein the extraction temperature is 80-85 ℃, and the extraction time is 2.0 hours; and filtering, then carrying out secondary extraction, controlling the water addition amount to be 10 times of the mass of the initial raw materials, controlling the extraction temperature to be 80-85 ℃, extracting for 1.5h, filtering, combining the filtrates, heating to 100 ℃, and carrying out evaporation concentration until the total weight of the plant raw materials is 28-35%.
(3) Adding 95% ethanol into the concentrate until the ethanol content is 70%, uniformly stirring, standing for 24h, taking the supernatant to a rotary evaporation device, evaporating the ethanol to obtain a semi-solidified viscous substance, and mixing the semi-solidified viscous substance with the ethanol according to the mass ratio of 1: adding water according to the proportion of 1, mixing uniformly, adding 0.2 of the pseudo-rue-odorous grass flower essential oil, 2.0 of the compound sodium nitrophenolate and 0.025 of the OP-10 emulsifier according to the mass proportion of the plant raw materials, and fully homogenizing by a homogenizer to obtain the invention.
0.35kg of the plant protective agent of the embodiment is diluted by water by 300 times, and is uniformly sprayed to 1 mu of citrus plants infected with the salpingosis, and the salpingosis germs are inhibited after 30 days of application, which shows that the plant bactericide of the embodiment has good control effect on the salpingosis of citrus.
The plant bactericide has the bacteriostatic and bactericidal effects on ralstonia solanacearum, citrus shaggy skin pathogenic bacteria and huanglongbing pathogenic bacteria:
firstly, taking the plant bactericide of example 1 as an example, the indoor toxicity of the plant bactericide of the invention to ralstonia solanacearum is measured:
adding the plant protective agent in the embodiment 1 into the PDA culture solution, mixing to prepare samples with plant protective agent concentrations of 18.75 mug/g, 37.50 mug/g, 75.00 mug/g, 150 mug/g, 750 mug/g, 1500 mug/g and 3000 mug/g respectively, sterilizing to obtain PDA culture media containing plant protective agents with different concentrations, and adopting the PDA culture media as a control; inoculating purified ralstonia solanacearum on a TTC culture medium, culturing in a 37 ℃ incubator, and beating a plurality of fungus cakes from the edges of colonies by using a puncher with the diameter of 0.4cm for later use; the bacterial cakes are transplanted to a PDA culture medium and a TTC culture medium containing plant bactericides with different concentrations reversely by using an inoculating needle, 3 bacterial cakes are arranged in each dish, the treatment is repeated twice, the bacterial cakes are placed in an incubator at 37 ℃ for 24 hours, then the diameter of the bacterial colonies is measured by using a microscope with a scale, the obtained data is processed by using EXCEL software, the relative inhibition rate and the probability value are obtained, the logarithm of the concentration is used as the horizontal coordinate (X), the probability value is used as the vertical coordinate (Y), a regression equation is obtained by scattered point regression analysis, and the logarithm of the concentration of EC50 is obtained when the Y is 0, and the result is shown in Table 1.
TABLE 1 indoor toxicity test results of plant bactericide against Ralstonia solanacearum
As can be seen from table 1, the plant fungicide of the present invention has EC50 of 207.04 μ g/g against ralstonia solanacearum, the regression equation of virulence is 1.3219x-3.0616, and the correlation coefficient R is 0.987, which indicates that the plant fungicide of the present invention has strong inhibitory activity against ralstonia solanacearum.
Taking the plant bactericide of the embodiment 1 as an example, measuring the indoor toxicity of the plant bactericide of the invention to the citrus sandiasis pathogenic bacteria:
the plant bactericide of example 1 was formulated into seven concentration gradient solutions of 18.75. mu.g/g, 37.50. mu.g/g, 75.00. mu.g/g, 150. mu.g/g, 375. mu.g/g, 750. mu.g/g and 1500. mu.g/g, and after sterilization, the sporangium on the back side of citrus russiang leaf was scraped thereinto, and an equal amount of the sporangium of citrus russiang pathogen was added to the sterilized water as a control. After the moisture-preserving culture is carried out in a large culture dish for 24 hours at room temperature, the number of germinated spores and the total number of spores in a visual field are observed under a microscope, the obtained data are processed by EXCEL software to obtain a relative inhibition rate and a probability value, a regression equation is obtained by scattered point regression analysis by taking a concentration logarithm as a horizontal coordinate (X) and a probability value as a vertical coordinate (Y), and the concentration logarithm of EC50 is obtained when Y is 0, wherein the results are shown in Table 2.
Corrected inhibition rate ═ number of blank control germinated spores-number of germinated spores in sample group)/number of blank control germinated spores
TABLE 2 indoor toxicity test results of plant bactericide for pathogenic bacteria of citrus sand disease
As can be seen from Table 2, the EC50 of the plant protective agent of the invention on the pathogenic bacteria of citrus salpingosis is 34.77 mu g/g, the toxicity regression equation is Y1.0995 x-1.6946, and the correlation coefficient R is 0.9334, which shows that the plant protective agent of the invention has stronger inhibitory activity on the pathogenic bacteria of citrus salpingosis.
Thirdly, taking the plant bactericide of the embodiment 1 as an example, the control effect of the plant bactericide on the pathogenic bacteria of the citrus greening disease is measured
Selecting citrus trees obviously infected with yellow dragon disease, taking about 30d of fresh leaves, storing with liquid nitrogen, spraying 200 times of diluent of the plant bactericide, and taking leaves with picked symmetrical parts after 20d, wherein each group comprises 3 parallel leaves. Then, carrying out PCR amplification by using a specific primer fA2/rJ5 of a beta-operon of an Asia bacterial system of the citrus greening disease, obtaining a standard curve of the fluorescence quantitative detection of the greening disease through the fluorescence quantitative PCR detection analysis, and calculating the bacterial content by using a regression equation, wherein the obtained result is shown in the following table 3:
TABLE 3 prevention and treatment effect of plant bactericide on pathogenic bacteria of citrus greening disease
Note: each group of three parallel
As can be seen from Table 3, the 20d inhibition rate of the plant bactericide of the invention on the pathogenic bacteria of citrus greening disease can reach 77.02%, although the plant bactericide can not completely kill the pathogenic bacteria of citrus greening disease, the plant bactericide still shows strong inhibition capability, and the plant bactericide of the invention has good inhibition activity on the pathogenic bacteria of citrus greening disease.
In conclusion, the plant source related to the invention is easy to obtain, the preparation process is simple, the cost is low, and the method is suitable for agricultural/forestry industrial production; the plant bactericide has stronger killing capability to ralstonia solanacearum and citrus sand-skin disease and also has certain inhibition effect to pathogenic bacteria of yellow dragon disease; the invention has no toxicity to human body, the dosage form is emulsion, the use is safe, the plant is easy to absorb after spraying, the invention has no pollution to the environment, and the invention has strong popularization and application value.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, changes and equivalent structural changes made to the above embodiment according to the technical spirit of the present invention still fall within the protection scope of the technical solution of the present invention.
Claims (4)
1. The preparation method of the plant compound bactericide is characterized by being prepared from the following raw materials in percentage by mass: stem and leaf of pseudo-osbeckia, ginkgo leaf, houttuynia, scutellaria, isatis root, forsythia, andrographis, sodium nitrophenolate, OP-10 emulsifier, pseudo-osbeckia flower essential oil and water, wherein the weight ratio of the stem and leaf of pseudo-osbeckia is 20-40, 2-5: 4-16: 2-6: 3-8: 2-8: 1.0-2.5: 0.01-0.03: 0.1-0.25: 10-30;
the method comprises the following steps:
(1) collecting the pseudo-osmyl grass, separately placing the flowers and the stems and leaves, airing the flowers in a shade place, and then placing the pseudo-osmyl grass flowers in an evaporation-condensation container/equipment according to a mass ratio of 1: adding water at a ratio of material to liquid of 20, evaporating at 100 ℃, and collecting condensate to obtain the essential oil of the pseudo-osbeckia chinensis;
(2) firstly, cutting plant raw materials into pieces, cutting ginkgo leaves, houttuynia cordata and stem leaves of pseudo-rue foetida into strips of 1-2 cm, cutting other wood raw materials into 1cm stem segments, uniformly mixing the cut wood raw materials in proportion, pouring the mixture into a reaction kettle, adding water which is 15-20 times of the mass of the plant raw materials, stirring and extracting, wherein the extraction temperature is 80-85 ℃, and the extraction time is 1.5-2.0 hours; filtering, then carrying out secondary extraction, controlling the water addition amount to be 8-12 times of the mass of the initial plant raw material, controlling the extraction temperature to be 80-85 ℃, extracting for 1-1.5 h, filtering, then combining the filtrates, heating to 100 ℃, and carrying out evaporation concentration until the total weight of the plant raw material is 28-35%;
(3) adding 95% ethanol into the concentrate obtained in the step (2) until the mass percentage of the ethanol is 60% -70%, uniformly stirring, standing for 16-24 h, taking the supernatant to rotary evaporation equipment, evaporating the ethanol to obtain a semi-solidified/viscous substance, and then mixing the semi-solidified/viscous substance according to the mass ratio of 1: adding water in a ratio of 0.8-1.2, mixing uniformly, adding 0.1-0.25 mass percent of pseudo-odorous grass flower essential oil, 1.0-2.5 mass percent of compound sodium nitrophenolate and 0.01-0.03 mass percent of OP-10 emulsifier continuously according to the mass ratio of the plant raw materials, and fully homogenizing by a homogenizer to obtain the plant compound bactericide.
2. A plant compound bactericide, which is prepared by the preparation method of claim 1.
3. The use of the compound plant bactericide according to claim 2, for killing ralstonia solanacearum and sorgo citrea.
4. The use of a plant compound bactericide as claimed in claim 2, for inhibiting huanglongbing pathogen.
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| CN108624527A (en) * | 2018-05-12 | 2018-10-09 | 湖南科技学院 | A kind of ginkgo source growth-promoting preparation of prevention ginger bacterial wilt |
| CN111567525A (en) * | 2020-05-08 | 2020-08-25 | 易之泰生物科技（龙岩）有限公司 | Composition and method for preventing and treating citrus greening disease |
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