Preparation method and agricultural application of traditional Chinese medicine composition

Abstract

The invention relates to the technical field of agricultural fertilizers, and discloses a preparation method and agricultural application of a traditional Chinese medicine composition. According to the preparation method of the traditional Chinese medicine composition provided by the invention, 25 traditional Chinese medicines are taken as raw materials, and are extracted, separated and purified to obtain the traditional Chinese medicine composition. The traditional Chinese medicine composition prepared by the preparation method of the traditional Chinese medicine composition provided by the invention can effectively increase the chlorophyll content of plants and effectively enhance the PPO activity and POD activity of the plants, so that the disease and pest resistance and stress resistance of the plants are effectively enhanced.

Description

Technical Field

[0001] The present invention relates to the technical field of agricultural fertilizers, and in particular to a preparation method and agricultural application of a traditional Chinese medicine composition. Background Art

[0002] Enhancing the resistance of plants to pests and diseases and stress is crucial for agricultural production and crop health. For example, enhancing the resistance of plants to pests and diseases and stress can significantly reduce the use of pesticides, thereby protecting the ecological environment and human health; enhancing the resistance of plants to pests and diseases and stress can enable crops to grow normally in adverse environments, thereby ensuring the yield and quality of crops; enhancing the resistance of plants to pests and diseases and stress can help crops adapt to extreme environmental conditions and reduce losses caused by natural disasters. Based on this, providing a Chinese medicine composition that can effectively enhance the resistance of plants to pests and diseases and stress has become a technical problem that needs to be solved urgently by those skilled in the art. Summary of the invention

[0003] The purpose of the present invention is to provide a preparation method and agricultural application of a Chinese medicine composition, which can effectively enhance the disease and insect pest resistance and stress resistance of plants.

[0004] To achieve the above object, the present invention provides the following technical solutions:

[0005] One of the technical solutions of the present invention:

[0006] A method for preparing a Chinese medicine composition comprises the following steps:

[0007] 1) mixing and grinding kelp, pine needles, stemona, sophora flavescens, kelp, sophora japonica leaves, ginkgo leaves, purslane, hawthorn leaves, capillaris and plantain to obtain a first group of Chinese medicine powders;

[0008] 2) using 70% ethanol as an extractant to perform reflux extraction on the first group of Chinese medicine powders obtained in step 1), and then extracting the extract with ethyl acetate and n-butanol in sequence to obtain an ethyl acetate portion and an n-butanol portion;

[0009] 3) applying the ethyl acetate obtained in step 2) to a silica gel column and performing gradient elution using a dichloromethane-methanol solution as an eluent to obtain components f-E1 to f-E4;

[0010] 4) mixing tomato stalks, Artemisia annua, dried orange peel, Rubescens rubescens, Lygodium japonicum, Houttuynia cordata, Polygonum cuspidatum, walnut peel, Licorice, Equisetum arvense, Viola yedoensis, Pueraria root, Clove and Wolfberry chamaejasme, and grinding to obtain a second group of Chinese medicine powders;

[0011] 5) using water as an extractant to perform reflux extraction on the second group of Chinese medicine powders obtained in step 4), and then extracting the extract with petroleum ether, chloroform and n-butanol in sequence to obtain a petroleum ether portion, a chloroform portion and an n-butanol portion;

[0012] 6) applying the petroleum ether obtained in step 5) to a silica gel column and performing gradient elution using a petroleum ether-acetone solution as an eluent to obtain components s-PE-E1 to s-PE-E6;

[0013] 7) applying the chloroform portion obtained in step 5) to an MCI gel column and performing gradient elution using a methanol solution as an eluent to obtain components s-Ch-E1 to s-Ch-E5;

[0014] 8) f-E3 obtained in step 3), s-PE-E2 obtained in step 6), and s-Ch-E4 obtained in step 7) are subjected to reduced pressure distillation to remove the eluent to obtain solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4; then, the solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4 obtained by reduced pressure distillation are mixed to obtain the Chinese medicine composition.

[0015] Furthermore, in step 1), the mass ratio of kelp, pine needles, stemona, sophora flavescens, esculenta, sophora japonica leaves, ginkgo leaves, purslane, hawthorn leaves, capillaris and plantain is (30-50): (5-10): (5-10): (3-10): (10-20): (10-15): (6-10): (6-10): (2-5): (2-5): (2-5).

[0016] Furthermore, in step 2), the solid-liquid ratio of the reflux extraction is 1 g:15 mL, the temperature is 70° C., and the time is 2 h.

[0017] Furthermore, in step 3), the gradient of the dichloromethane-methanol solution is 0:5 (v:v), 1:5 (v:v), 3:5 (v:v) and 5:5 (v:v).

[0018] Further, in step 4), the mass ratio of the tomato stalk, Artemisia annua, tangerine peel, Rubescens rubescens, Lygodium japonicus, Houttuynia cordata, Polygonum cuspidatum, walnut peel, liquorice, Equisetum arvense, Viola yedoensis, Pueraria root, clove and Stellera chamaejasme is (10-20):(2-3):(4-8):(4-8):(4-8):(4-8):(8-15):(2-3):(8-15):(4-10):(4-10):(4-10):(8-10):(10-15).

[0019] Furthermore, in step 5), the solid-liquid ratio of the reflux extraction is 1 g:8 mL, the temperature is 90° C., and the time is 1 h.

[0020] Furthermore, in step 6), the gradient of the petroleum ether-acetone solution is 100:0 (v:v), 98:2 (v:v), 95:5 (v:v), 90:10 (v:v), 70:30 (v:v), and 50:50 (v:v).

[0021] Furthermore, in step 7), the gradient of the methanol solution is 0%, 20%, 50%, 90% and 100%.

[0022] Furthermore, in step 8), the mass ratio of the solid material f-E3, the solid material s-PE-E2 and the solid material s-Ch-E4 is 1:1:1.

[0023] The second technical solution of the present invention:

[0024] The Chinese medicine composition prepared by the above-mentioned preparation method of the Chinese medicine composition is used in enhancing the immune function, disease and pest resistance, and stress resistance of plants.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The Chinese medicine composition prepared by the preparation method of a Chinese medicine composition provided by the present invention can effectively increase the chlorophyll content of plants, and effectively enhance the PPO activity and POD activity of plants, thereby effectively enhancing the disease and insect pest resistance and stress resistance of plants. DETAILED DESCRIPTION

[0027] Now, various exemplary embodiments of the present invention are described in detail, and this detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present invention. It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention.

[0028] In addition, for the numerical range in the present invention, it is understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0029] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0030] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0031] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0032] In the following examples and comparative examples, all the raw materials and reagents used were commercially available.

[0033] In the following examples, a method for preparing a Chinese medicine composition comprises the following steps:

[0034] 1) mixing kelp, pine needles, stemona, sophora flavescens, kelp fusiformis, sophora japonica leaf, ginkgo leaf, purslane, hawthorn leaf, capillaris and plantain in a mass ratio of (30-50): (5-10): (5-10): (3-10): (10-20): (10-15): (6-10): (6-10): (2-5): (2-5): (2-5), grinding the mixture and passing through a 40-mesh sieve to obtain a first group of Chinese medicine powders;

[0035] 2) placing the first group of Chinese medicine powder obtained in step 1) in 70% ethanol at a material-liquid ratio of 1 g:15 mL, and extracting under reflux at 70° C. for 2 h, repeating 3 times, combining the extracts, and then extracting the extracts with ethyl acetate and n-butanol in sequence to obtain an ethyl acetate portion and an n-butanol portion;

[0036] 3) applying the ethyl acetate obtained in step 2) to a silica gel column and performing gradient elution using a dichloromethane-methanol solution as an eluent to obtain components f-E1 to f-E4;

[0037] The gradient of the dichloromethane-methanol solution is 0:5 (v:v), 1:5 (v:v), 3:5 (v:v) and 5:5 (v:v);

[0038] 4) mixing tomato stalks, artemisia annua, tangerine peel, rubescens herb, sea gold sand grass, houttuynia cordata, knotweed, walnut peel, licorice, aralia arvense, Viola yedoensis, kudzu root, clove and wolfsbane in a mass ratio of (10-20): (2-3): (4-8): (4-8): (4-8): (4-8): (8-15): (2-3): (8-15): (4-10): (4-10): (4-10): (8-10): (10-15), grinding and passing through a 40-mesh sieve to obtain a second group of Chinese medicine powders;

[0039] 5) placing the second Chinese medicine powder obtained in step 4) in water at a material-liquid ratio of 1 g:8 mL, and extracting under reflux at 90° C. for 1 h, repeating 3 times, combining the extracts, and then extracting the extracts with petroleum ether, chloroform and n-butanol in sequence to obtain a petroleum ether portion, a chloroform portion and a n-butanol portion;

[0040] 6) applying the petroleum ether obtained in step 5) to a silica gel column and performing gradient elution using a petroleum ether-acetone solution as an eluent to obtain components s-PE-E1 to s-PE-E6;

[0041] The gradient of petroleum ether-acetone solution is 100:0 (v:v), 98:2 (v:v), 95:5 (v:v), 90:10 (v:v), 70:30 (v:v), 50:50 (v:v);

[0042] 7) applying the chloroform portion obtained in step 5) to an MCI gel column and performing gradient elution using a methanol solution as an eluent to obtain components s-Ch-E1 to s-Ch-E5;

[0043] The gradient of the methanol solution is 0%, 20%, 50%, 90% and 100%;

[0044] 8) f-E3 obtained in step 3), s-PE-E2 obtained in step 6), and s-Ch-E4 obtained in step 7) are subjected to reduced pressure distillation to remove the eluent to obtain solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4; then, the solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4 obtained by reduced pressure distillation are mixed in a mass ratio of 1:1:1, ball-milled, and then pressed into spherical particles with a particle size of about 2 mm, i.e., the Chinese medicine composition.

[0045] Example 1

[0046] A Chinese medicine composition

[0047] 1) according to the mass ratio of kelp, pine needle, stemona, sophora flavescens, kelp, sophora japonica leaf, ginkgo leaf, purslane, hawthorn leaf, capillaris and plantain being 30:5:5:3:10:10:6:6:2:2:2, kelp, pine needle, stemona, sophora flavescens, kelp, sophora japonica leaf, ginkgo leaf, purslane, hawthorn leaf, capillaris and plantain are mixed, ground and passed through a 40-mesh sieve to obtain a first group of Chinese medicine powders;

[0048] 2) placing the first group of Chinese medicine powder obtained in step 1) in 70% ethanol at a material-liquid ratio of 1 g:15 mL, and extracting under reflux at 70° C. for 2 h, repeating 3 times, combining the extracts, and then extracting the extracts with ethyl acetate and n-butanol in sequence to obtain an ethyl acetate portion and an n-butanol portion;

[0049] 3) applying the ethyl acetate obtained in step 2) to a silica gel column and performing gradient elution using a dichloromethane-methanol solution as an eluent to obtain components f-E1 to f-E4;

[0050] The gradient of the dichloromethane-methanol solution is 0:5 (v:v), 1:5 (v:v), 3:5 (v:v) and 5:5 (v:v);

[0051] 4) mixing tomato stalks, artemisia annua, tangerine peel, rubescens herb, sea gold sand grass, houttuynia cordata, knotweed, walnut peel, licorice, aralia arvense, Viola yedoensis, kudzu root, clove and wolfsbane in a mass ratio of 10:2:4:4:4:4:8:2:8:4:4:4:8:10, grinding and passing through a 40-mesh sieve to obtain a second group of Chinese medicine powders;

[0052] 5) placing the second Chinese medicine powder obtained in step 4) in water at a material-liquid ratio of 1 g:8 mL, and extracting under reflux at 90° C. for 1 h, repeating 3 times, combining the extracts, and then extracting the extracts with petroleum ether, chloroform and n-butanol in sequence to obtain a petroleum ether portion, a chloroform portion and a n-butanol portion;

[0053] 6) applying the petroleum ether obtained in step 5) to a silica gel column and performing gradient elution using a petroleum ether-acetone solution as an eluent to obtain components s-PE-E1 to s-PE-E6;

[0054] The gradient of petroleum ether-acetone solution is 100:0 (v:v), 98:2 (v:v), 95:5 (v:v), 90:10 (v:v), 70:30 (v:v), 50:50 (v:v);

[0055] 7) applying the chloroform portion obtained in step 5) to an MCI gel column and performing gradient elution using a methanol solution as an eluent to obtain components s-Ch-E1 to s-Ch-E5;

[0056] The gradient of the methanol solution is 0%, 20%, 50%, 90% and 100%;

[0057] 8) f-E3 obtained in step 3), s-PE-E2 obtained in step 6), and s-Ch-E4 obtained in step 7) are subjected to reduced pressure distillation to remove the eluent to obtain solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4; then, the solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4 obtained by reduced pressure distillation are mixed in a mass ratio of 1:1:1, ball-milled, and then pressed into spherical particles with a particle size of about 2 mm, i.e., the Chinese medicine composition.

[0058] Example 2

[0059] A Chinese medicine composition

[0060] 1) according to the mass ratio of kelp, pine needle, stemona, sophora flavescens, kelp, sophora japonica leaf, ginkgo leaf, purslane, hawthorn leaf, capillaris and plantain being 40:8:8:7:16:12:8:8:4:4:4, kelp, pine needle, stemona, sophora flavescens, kelp, sophora japonica leaf, ginkgo leaf, purslane, hawthorn leaf, capillaris and plantain are mixed, ground and passed through a 40-mesh sieve to obtain a first group of Chinese medicine powders;

[0061] 2) placing the first group of Chinese medicine powder obtained in step 1) in 70% ethanol at a material-liquid ratio of 1 g:15 mL, and extracting under reflux at 70° C. for 2 h, repeating 3 times, combining the extracts, and then extracting the extracts with ethyl acetate and n-butanol in sequence to obtain an ethyl acetate portion and an n-butanol portion;

[0062] 3) applying the ethyl acetate obtained in step 2) to a silica gel column and performing gradient elution using a dichloromethane-methanol solution as an eluent to obtain components f-E1 to f-E4;

[0063] The gradient of the dichloromethane-methanol solution is 0:5 (v:v), 1:5 (v:v), 3:5 (v:v) and 5:5 (v:v);

[0064] 4) mixing tomato stalks, artemisia annua, tangerine peel, rubescens herb, sea gold sand grass, houttuynia cordata, knotweed, walnut peel, licorice, aralia arvense, Viola yedoensis, kudzu root, clove and wolfsbane in a mass ratio of 15:2.5:5:5:5:5:10:2.5:10:6:6:6:9:12, grinding and passing through a 40-mesh sieve to obtain a second group of Chinese medicine powders;

[0065] 5) placing the second Chinese medicine powder obtained in step 4) in water at a material-liquid ratio of 1 g:8 mL, and extracting under reflux at 90° C. for 1 h, repeating 3 times, combining the extracts, and then extracting the extracts with petroleum ether, chloroform and n-butanol in sequence to obtain a petroleum ether portion, a chloroform portion and a n-butanol portion;

[0066] 6) applying the petroleum ether obtained in step 5) to a silica gel column and performing gradient elution using a petroleum ether-acetone solution as an eluent to obtain components s-PE-E1 to s-PE-E6;

[0067] The gradient of petroleum ether-acetone solution is 100:0 (v:v), 98:2 (v:v), 95:5 (v:v), 90:10 (v:v), 70:30 (v:v), 50:50 (v:v);

[0068] 7) applying the chloroform portion obtained in step 5) to an MCI gel column and performing gradient elution using a methanol solution as an eluent to obtain components s-Ch-E1 to s-Ch-E5;

[0069] The gradient of the methanol solution is 0%, 20%, 50%, 90% and 100%;

[0070] 8) f-E3 obtained in step 3), s-PE-E2 obtained in step 6), and s-Ch-E4 obtained in step 7) are subjected to reduced pressure distillation to remove the eluent to obtain solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4; then, the solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4 obtained by reduced pressure distillation are mixed in a mass ratio of 1:1:1, ball-milled, and then pressed into spherical particles with a particle size of about 2 mm, i.e., the Chinese medicine composition.

[0071] Example 3

[0072] A Chinese medicine composition

[0073] 1) according to the mass ratio of kelp, pine needle, stemona, sophora flavescens, kelp, sophora japonica leaf, ginkgo leaf, purslane, hawthorn leaf, capillaris and plantain being 50:10:10:10:20:15:10:10:5:5:5, kelp, pine needle, stemona, sophora flavescens, kelp, sophora japonica leaf, ginkgo leaf, purslane, hawthorn leaf, capillaris and plantain are mixed, ground and passed through a 40-mesh sieve to obtain a first group of Chinese medicine powders;

[0074] 2) placing the first group of Chinese medicine powder obtained in step 1) in 70% ethanol at a material-liquid ratio of 1 g:15 mL, and extracting under reflux at 70° C. for 2 h, repeating 3 times, combining the extracts, and then extracting the extracts with ethyl acetate and n-butanol in sequence to obtain an ethyl acetate portion and an n-butanol portion;

[0075] 3) applying the ethyl acetate obtained in step 2) to a silica gel column and performing gradient elution using a dichloromethane-methanol solution as an eluent to obtain components f-E1 to f-E4;

[0076] The gradient of the dichloromethane-methanol solution is 0:5 (v:v), 1:5 (v:v), 3:5 (v:v) and 5:5 (v:v);

[0077] 4) mixing tomato stalks, artemisia annua, tangerine peel, rubescens herb, sea gold sand grass, houttuynia cordata, knotweed, walnut peel, licorice, aralia arvense, Viola yedoensis, kudzu root, clove and wolfsbane in a mass ratio of 20:3:8:8:8:8:15:3:15:10:10:10:10:15, grinding and passing through a 40-mesh sieve to obtain a second group of Chinese medicine powders;

[0078] 5) placing the second Chinese medicine powder obtained in step 4) in water at a material-liquid ratio of 1 g:8 mL, and extracting under reflux at 90° C. for 1 h, repeating 3 times, combining the extracts, and then extracting the extracts with petroleum ether, chloroform and n-butanol in sequence to obtain a petroleum ether portion, a chloroform portion and a n-butanol portion;

[0079] 6) applying the petroleum ether obtained in step 5) to a silica gel column and performing gradient elution using a petroleum ether-acetone solution as an eluent to obtain components s-PE-E1 to s-PE-E6;

[0080] The gradient of petroleum ether-acetone solution is 100:0 (v:v), 98:2 (v:v), 95:5 (v:v), 90:10 (v:v), 70:30 (v:v), 50:50 (v:v);

[0081] 7) applying the chloroform portion obtained in step 5) to an MCI gel column and performing gradient elution using a methanol solution as an eluent to obtain components s-Ch-E1 to s-Ch-E5;

[0082] The gradient of the methanol solution is 0%, 20%, 50%, 90% and 100%;

[0083] 8) f-E3 obtained in step 3), s-PE-E2 obtained in step 6), and s-Ch-E4 obtained in step 7) are subjected to reduced pressure distillation to remove the eluent to obtain solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4; then, the solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4 obtained by reduced pressure distillation are mixed in a mass ratio of 1:1:1, ball-milled, and then pressed into spherical particles with a particle size of about 2 mm, i.e., the Chinese medicine composition.

[0084] Comparative Example

[0085] A Chinese medicine composition

[0086] 1) according to the mass ratio of kelp, pine needle, stemona, sophora flavescens, kelp, sophora japonica leaf, ginkgo leaf, purslane, hawthorn leaf, capillaris and plantain being 40:8:8:7:16:12:8:8:4:4:4, kelp, pine needle, stemona, sophora flavescens, kelp, sophora japonica leaf, ginkgo leaf, purslane, hawthorn leaf, capillaris and plantain are mixed, ground and passed through a 40-mesh sieve to obtain a first group of Chinese medicine powders;

[0087] 2) placing the first group of Chinese medicine powders obtained in step 1) in 70% ethanol at a material-liquid ratio of 1 g:15 mL, and extracting under reflux at 70° C. for 2 h, repeating the process three times, combining the extracts, concentrating and drying the extracts, and obtaining the first group of Chinese medicine extracts;

[0088] 3) mixing tomato stalks, artemisia annua, tangerine peel, rubescens herb, sea gold sand grass, houttuynia cordata, knotweed, walnut peel, licorice, aralia arvense, Viola yedoensis, kudzu root, clove and wolfsbane in a mass ratio of 15:2.5:5:5:5:5:10:2.5:10:6:6:6:9:12, grinding and passing through a 40-mesh sieve to obtain a second group of Chinese medicine powders;

[0089] 4) placing the second Chinese medicine powder obtained in step 4) in water at a material-liquid ratio of 1 g:8 mL, and extracting under reflux at 90° C. for 1 h, repeating 3 times, combining the extracts, concentrating and drying the extracts, and obtaining a second group of Chinese medicine extracts;

[0090] 5) The first group of Chinese herbal extracts obtained in step 2) and the second group of Chinese herbal extracts obtained in step 4) are mixed at a mass ratio of 1:1, ball-milled, and then pressed into spherical particles with a particle size of about 2 mm, i.e., the Chinese herbal composition.

[0091] Effect verification

[0092] Tomatoes were sown in nutrient pots for seedling raising. When the tomato seedlings grew to the stage of four leaves and one heart, 200 tomato seedlings with the same growth were selected and randomly divided into 5 groups.

[0093] Experimental group I: the Chinese medicinal composition prepared in Example 1 was diluted according to a mass volume ratio of 1 g: 2 L, and then the diluted Chinese medicinal composition was used to irrigate the roots of tomato seedlings;

[0094] Experimental Group II: The Chinese medicinal composition prepared in Example 2 was diluted according to a mass volume ratio of 1 g: 2 L, and then the diluted Chinese medicinal composition was used to irrigate the roots of tomato seedlings;

[0095] Experimental group III: the Chinese medicinal composition prepared in Example 3 was diluted according to a mass volume ratio of 1 g: 2 L, and then the diluted Chinese medicinal composition was used to irrigate the roots of tomato seedlings;

[0096] Control group: dilute the Chinese medicinal composition prepared in the comparative example according to a mass volume ratio of 1 g: 2 L, and then use the diluted Chinese medicinal composition to irrigate the roots of tomato seedlings;

[0097] Blank control group: the tomato seedlings were irrigated with an equal volume of water;

[0098] The seedlings were raised under simulated natural conditions for 15 days, and then the chlorophyll content of the leaves of the same parts of the tomato seedlings in the experimental group I, experimental group II, experimental group III, control group and blank control group was detected using a SPAD-502 handheld chlorophyll meter. The chlorophyll content detection results are shown in Table 1;

[0099] Table 1 Chlorophyll content test results (average value)

[0100]

[0101]

[0102] It can be seen from the data in Table 1 that the Chinese medicine composition prepared by the preparation method of a Chinese medicine composition provided by the present invention can effectively increase the chlorophyll content of plants; studies have shown that there is a positive correlation between the chlorophyll content in plants and the photosynthesis efficiency of plants. When the chlorophyll content in plants increases, the plants can more effectively capture and utilize light energy, improve photosynthesis efficiency, and promote the accumulation of nutrients in plants. Moreover, the increase in chlorophyll content can promote the synthesis of amino acids and nucleic acids in plants and their absorption, conduction, and metabolic capabilities, improve the immune function of plants, and enhance the resistance of plants to diseases and pests and stress. Based on this, the Chinese medicine composition prepared by the preparation method of a Chinese medicine composition provided by the present invention can effectively enhance the resistance to diseases and pests and stress of plants.

[0103] Referring to "The induction of salicylic acid and acetylsalicylic acid on PPO and POD in tomato seedling leaves", Hu Yanjiang et al. tested the PPO activity and POD activity of leaves of tomato seedlings at the same position in experimental group I, experimental group II, experimental group III, control group and blank control group. The results of the PPO activity and POD activity tests are shown in Table 2.

[0104] Table 2 PPO activity and POD activity test results

[0105]

[0106] It can be seen from the data in Table 2 that the Chinese medicine composition prepared by the method for preparing a Chinese medicine composition provided by the present invention can effectively enhance the PPO activity and POD activity of the plant; PPO and POD can promote the synthesis rate of secondary metabolites of the plant. Therefore, the increase in PPO activity and POD activity in the plant can effectively increase the content of secondary metabolites in the plant. The secondary metabolites of the plant and their degradation products play an important role in enhancing the disease and pest resistance and stress resistance of the plant. Based on this, the Chinese medicine composition prepared by the method for preparing a Chinese medicine composition provided by the present invention can effectively enhance the disease and pest resistance and stress resistance of the plant.

[0107] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in the relevant field should understand that the specific implementation methods of the present invention can still be modified or replaced by equivalents. Any modification or equivalent replacement that does not depart from the spirit and scope of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. A method for preparing a Chinese medicine composition, characterized in that: The following steps are involved: 1) mixing and grinding kelp, pine needles, stemona, sophora flavescens, kelp, sophora japonica leaves, ginkgo leaves, purslane, hawthorn leaves, capillaris and plantain to obtain a first group of Chinese medicine powders; 2) using 70% ethanol as an extractant to perform reflux extraction on the first group of Chinese medicine powders obtained in step 1), and then extracting the extract with ethyl acetate and n-butanol in sequence to obtain an ethyl acetate portion and an n-butanol portion; 3) applying the ethyl acetate obtained in step 2) to a silica gel column and performing gradient elution using a dichloromethane-methanol solution as an eluent to obtain components f-E1 to f-E4; 4) mixing tomato stalks, Artemisia annua, dried orange peel, Rubescens rubescens, Lygodium japonicum, Houttuynia cordata, Polygonum cuspidatum, walnut peel, Licorice, Equisetum arvense, Viola yedoensis, Pueraria root, Clove and Wolfberry chamaejasme, and grinding to obtain a second group of Chinese medicine powders; 5) using water as an extractant to perform reflux extraction on the second group of Chinese medicine powders obtained in step 4), and then extracting the extract with petroleum ether, chloroform and n-butanol in sequence to obtain a petroleum ether portion, a chloroform portion and an n-butanol portion; 6) applying the petroleum ether obtained in step 5) to a silica gel column and gradient eluting with a petroleum ether-acetone solution as an eluent to obtain components s-PE-E1 to s-PE-E6; 7) applying the chloroform portion obtained in step 5) to an MCI gel column and performing gradient elution using a methanol solution as an eluent to obtain components s-Ch-E1 to s-Ch-E5; 8) f-E3 obtained in step 3), s-PE-E2 obtained in step 6), and s-Ch-E4 obtained in step 7) are subjected to reduced pressure distillation to remove the eluent to obtain solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4; then, the solid matter f-E3, solid matter s-PE-E2, and solid matter s-Ch-E4 obtained by reduced pressure distillation are mixed to obtain the Chinese medicine composition.

2. The method for preparing a Chinese medicine composition according to claim 1, characterized in that: In step 1), the mass ratio of kelp, pine needles, stemona, sophora flavescens, fusca, sophora japonica leaves, ginkgo leaves, purslane, hawthorn leaves, capillaris and plantain is (30-50): (5-10): (5-10): (3-10): (10-20): (10-15): (6-10): (6-10): (2-5): (2-5): (2-5).

3. The method for preparing a Chinese medicine composition according to claim 1, characterized in that: In step 2), the solid-liquid ratio of the reflux extraction is 1 g:15 mL, the temperature is 70° C., and the time is 2 h.

4. The method for preparing a Chinese medicine composition according to claim 1, characterized in that: In step 3), the gradient of the dichloromethane-methanol solution is 0:5 (v:v), 1:5 (v:v), 3:5 (v:v) and 5:5 (v:v).

5. The method for preparing a Chinese medicine composition according to claim 1, characterized in that: In step 4), the mass ratio of the tomato stalk, Artemisia annua, tangerine peel, Rubescens rubescens, Lygodium japonicus, Houttuynia cordata, Polygonum cuspidatum, walnut peel, liquorice, Equisetum arvense, Viola yedoensis, Pueraria root, clove and Stellera chamaejasme is (10-20):(2-3):(4-8):(4-8):(4-8):(4-8):(8-15):(2-3):(8-15):(4-10):(4-10):(4-10):(8-10):(10-15).

6. The method for preparing a Chinese medicine composition according to claim 1, characterized in that: In step 5), the solid-liquid ratio of the reflux extraction is 1 g:8 mL, the temperature is 90° C., and the time is 1 h.

7. The method for preparing a Chinese medicine composition according to claim 1, characterized in that: In step 6), the gradient of the petroleum ether-acetone solution is 100:0 (v:v), 98:2 (v:v), 95:5 (v:v), 90:10 (v:v), 70:30 (v:v), and 50:50 (v:v).

8. The method for preparing a Chinese medicine composition according to claim 1, characterized in that: In step 7), the gradient of the methanol solution is 0%, 20%, 50%, 90% and 100%.

9. The method for preparing a Chinese medicine composition according to claim 1, characterized in that: In step 8), the mass ratio of the solid material f-E3, the solid material s-PE-E2 and the solid material s-Ch-E4 is 1:1:

1.

10. Use of a Chinese medicine composition prepared by the method for preparing a Chinese medicine composition according to any one of claims 1 to 9 in enhancing the immune function, disease and pest resistance, and stress resistance of plants.

Images