Application of xanthocyanin in the preparation of plant-based herbicides and plant-based herbicides

Abstract

This invention discloses the application of zanthoxylin in the preparation of plant-based herbicides and the resulting herbicides, relating to the field of herbicide technology. The concentration of zanthoxylin in the plant-based herbicides is ≥20 μg / mL. By using zanthoxylin as the active ingredient in plant-based herbicides, it exhibits significant growth inhibition against various common farmland weeds at a concentration ≥20 μg / mL. Furthermore, this active ingredient is derived from natural plants and is a biodegradable secondary metabolite, making it less likely to leave residues in the environment and causing no soil degradation or water pollution. Compared to chemically synthesized herbicides, it is more ecologically safe. This not only provides a new option for developing efficient, broad-spectrum, and environmentally friendly bio-based herbicides but also expands the application prospects of zanthoxylin in the agricultural field.

Description

Technical Field

[0001] This invention relates to the field of weed control product technology, and more specifically, to the application of apigenin in the preparation of plant weed control products and plant herbicides. Background Technology

[0002] Chemically synthesized herbicides, such as glyphosate, paraquat, and atrazine, are widely used in agricultural production to effectively control weed growth and increase crop yields. These herbicides typically achieve selective or non-selective weed control by interfering with the physiological and biochemical processes of weeds. In modern agricultural technology, the use of chemical herbicides has become one of the important means of farmland management.

[0003] However, the long-term and widespread use of synthetic chemical herbicides has gradually had adverse effects on the ecological environment. On the one hand, weed resistance continues to evolve, leading to decreased efficacy, increased dosage, and higher application frequency, thus creating a vicious cycle of "increased resistance - increased dosage." On the other hand, residual chemical herbicides accumulate in soil and water, easily causing adverse effects on both the ecosystem and human health. Therefore, there is an urgent need to provide a new type of herbicide that is naturally derived, environmentally compatible, easily degradable, and possesses clear target activity to meet the planting needs of green agriculture, organic agriculture, and sustainable development of ecological farmland.

[0004] In view of this, the present invention is proposed. Summary of the Invention

[0005] The purpose of this invention is to provide an application of xanthocyanin in the preparation of plant herbicides and a plant herbicide.

[0006] This invention is implemented as follows: In a first aspect, the present invention provides an application of xanthocyanin in the preparation of plant herbicides, wherein the concentration of xanthocyanin in the plant herbicides is ≥20 μg / mL.

[0007] Secondly, the present invention provides a plant herbicide, wherein the active substance includes xanthocyanin, and the concentration of xanthocyanin in the plant herbicide product is ≥20 μg / mL, preferably 20~1000 μg / mL, more preferably 100~1000 μg / mL.

[0008] The present invention has the following beneficial effects: This invention provides an application of sanshool in the preparation of plant-based herbicides and a plant-based herbicide. By using sanshool as the active ingredient in the plant-based herbicide, it exhibits significant growth inhibition against various common farmland weeds at a concentration of ≥20 μg / mL. Moreover, this active ingredient is derived from natural plants and is a biodegradable secondary metabolite, which is not easily left in the environment and does not cause soil degradation or water pollution. Compared with chemically synthesized herbicides, it is more ecologically safe. This not only provides a new option for developing efficient, broad-spectrum, and environmentally friendly bio-based herbicides but also expands the application prospects of sanshool in the agricultural field. Attached Figure Description

[0009] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0010] Figure 1 The 1H NMR spectrum of xanthocyanin provided in this embodiment of the invention; Figure 2 The carbon NMR spectrum of xanthocyanin provided in this embodiment of the invention; Figure 3 This is a flowchart illustrating the separation process of xanthocyanin provided in an embodiment of the present invention; Figure 4 This is a diagram illustrating the effect of xanthocyanin on root length during the growth of four weeds, provided in an embodiment of the present invention. Figure 5 This is a diagram illustrating the effect of xanthocyanin on seedling height of four weeds during the growth process, provided in an embodiment of the present invention. Figure 6 The diagram illustrates the effect of xanthocyanin on alfalfa growth, as provided in an embodiment of the present invention. Figure 7 The diagram shows the effect of xanthocyanin on the growth of Amaranthus retroflexus, as provided in this embodiment of the invention. Figure 8 A visualization of the molecular docking between xanthophyll and Arabidopsis thaliana protoporphyrinogen IX oxidase PPO (1SEZ) provided in an embodiment of the present invention; Figure 9 This is a two-dimensional schematic diagram of the interaction between protein (Arabidopsis protoporphyrinogen IX oxidase PPO(1SEZ)) and ligand (piperazine). Detailed Implementation

[0011] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0012] In a first aspect, the present invention provides an application of xanthocyanin in the preparation of plant herbicides, wherein the concentration of xanthocyanin in the plant herbicides is ≥20 μg / mL.

[0013] In an optional embodiment, the concentration of xanthocyanin in the herbicide product is 20~1000 μg / mL.

[0014] In an optional embodiment, the concentration of xanthocyanin in the herbicide product is 100~1000 μg / mL.

[0015] By using xanthocyanin as the active ingredient in plant-based herbicides, it has shown significant growth inhibition against a variety of common farmland weeds at a concentration of ≥20 μg / mL. Moreover, the inhibitory effect increases with increasing concentration within a certain range. The preferred concentration of xanthocyanin in plant-based herbicides is 100~1000 μg / mL.

[0016] In an optional implementation, xanthocyanin achieves its weed-control effect by reducing the seedling length of weeds, reducing the root length of weeds, causing the leaves to become scorched, causing the leaves to develop chlorotic white spots, or causing the weeds to fall over.

[0017] In an optional implementation, the plant-based herbicide targets weeds including Amaranthus retroflexus (…). Amaranthus retroflexus ), foxtail grass ( Setaria viridis ), alfalfa Medicago sativa ) and gray-green quinoa ( Oxybasis glauca At least one of the following.

[0018] In an optional embodiment, xanthocyanin is an active substance extracted from Artemisia argyi in Yili that has the following structure: .

[0019] The cinnamon used in this invention is derived from natural plants and is a biodegradable secondary metabolite. It does not easily leave residues in the environment and does not cause soil degradation or water pollution. Compared with chemically synthesized herbicides, it is more ecologically safe. This invention not only provides a new option for developing efficient, broad-spectrum, and environmentally friendly bio-based herbicides, but also expands the application prospects of cinnamon in the agricultural field.

[0020] In an optional embodiment, the method for preparing xanthocyanin includes drying and pulverizing Artemisia argyi to obtain Artemisia argyi powder; extracting the Artemisia argyi powder and evaporating under reduced pressure to obtain Artemisia argyi extract; and obtaining xanthocyanin by gradient elution and chromatographic separation of the Artemisia argyi extract.

[0021] In an optional embodiment, the solvent used for extracting dried Artemisia argyi powder is 95% ethanol.

[0022] And / or, the reduced pressure evaporation temperature is 40~50℃.

[0023] And / or, the solvents used for gradient elution include petroleum ether and ethyl acetate.

[0024] And / or, chromatographic separation includes separation using column chromatography and / or high-performance liquid chromatography.

[0025] Secondly, the present invention provides a plant herbicide, wherein the active substance includes xanthocyanin, and the concentration of xanthocyanin in the plant herbicide product is ≥20 μg / mL, preferably 20~1000 μg / mL, more preferably 100~1000 μg / mL.

[0026] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0027] Example 1 This embodiment provides an application of xanthocyanin in the preparation of plant-based herbicides. The specific experimental method is as follows: 1. Extraction of xanthocyanin 1.1. Collection of Artemisia argyi (Ili Artemisia) Seriphidium transiliense The plant was dried at room temperature in a cool place in the laboratory and then ground into powder (total 3.0 kg).

[0028] 1.2. 3.0 kg of Artemisia argyi plant powder was extracted three times with 95% ethanol at room temperature. The extract was evaporated under reduced pressure at 45°C to obtain 249.94 g of Artemisia argyi ethanol extract.

[0029] 1.3. 249.94 g of Artemisia argyi ethanol extract was subjected to silica gel column chromatography with gradient elution. The volume ratio of petroleum ether to ethyl acetate in the eluent was 1:0 → 0:1, and the silica gel in the column had a particle size of 200-300 mesh. The fractions obtained after gradient elution were developed by TLC and anisaldehyde-sulfuric acid solution. The first crude product was obtained by combining the eluted fractions with the same color shift value in the thin-layer chromatography.

[0030] 1.4 The first crude product was treated with a Sephadex LH-20 column (dichloromethane / methanol ratio of 1:1) to obtain the second crude product. The second crude product was then further separated by preparative high performance liquid chromatography (methanol:water 90:10) to obtain 16.2 mg of pale yellow crystalline compound.

[0031] 1.5 The structure of the obtained pale yellow crystalline compound was determined using an XTIPC Varian MR-400 nuclear magnetic resonance spectrometer. 1 H NMR and 13 C NMR spectra, to obtain such Figure 1 and Figure 2 The results are shown. (From...) Figure 1 and Figure 2 The structural formula of the obtained pale yellow crystalline compound is as follows: This refers to xanthocyanin; the separation flowchart of xanthocyanin is shown below. Figure 3 As shown.

[0032] 2. Bioassay of herbicidal activity in petri dishes 2.1 Plant seed pretreatment: Pretreatment of Amaranthus retroflexus (…) Amaranthus retroflexus ), foxtail grass ( Setaria viridis ), alfalfa Medicago sativa ) and gray-green quinoa ( Oxybasis glauca The seeds were surface disinfected by soaking in 75% (v / v) ethanol for 5 minutes, and then rinsed three times with sterile distilled water for later use.

[0033] 2.2 Dissolve the xanthocyanin obtained in step 1 in 0.1% (v / v) methanol to prepare a xanthocyanin stock solution of 10 mg / mL. Then, dilute the xanthocyanin stock solution with distilled water to obtain xanthocyanin solutions with final concentrations of 20 μg / mL, 50 μg / mL, 100 μg / mL, 200 μg / mL, 500 μg / mL and 1000 μg / mL.

[0034] A 0.1% (v / v) methanol aqueous solution was used as the negative control solution, i.e., the concentration of xanthocyanin in the negative control solution was 0 μg / mL.

[0035] 2.3. Use 9cm diameter glass petri dishes, each lined with a layer of sterile filter paper. Add 4 mL of different concentrations of xanthocyanin solution to each petri dish, with 4 mL of negative control solution added to the negative control group. After adding the solution, evenly place 30 seeds pretreated in step 2.1 into each dish. Seal the petri dishes with plastic bags to maintain humidity and incubate at 25 ± 1℃ in the dark for 120 hours. Set up three independent replicates (i.e., three petri dishes) for each concentration. After incubation, measure the length of the plumule and radicle of each seed.

[0036] The effects of different concentrations (0 µg / mL, 20 µg / mL, 50 µg / mL, 100 µg / mL, 200 µg / mL, 500 µg / mL, and 1000 µg / mL) of xanthophyll on the growth of Amaranthus retroflexus, Setaria viridis, Alfalfa, and Chenopodium album were investigated, and the results were obtained as follows: Figure 4 and Figure 5 The results are shown.

[0037] Depend on Figure 4 and Figure 5 It was found that xanthocyanin at 100 μg / mL significantly inhibited the root length and seedling height of common weeds such as foxtail grass, lambsquarters, amaranth, and alfalfa seedlings; within the concentration range of 20–500 μg / mL, the inhibitory effect of xanthocyanin on the four weeds was significantly enhanced with increasing treatment concentration; when the concentration of xanthocyanin was 1000 μg / mL, it showed significant inhibition on the growth activity of the four weeds.

[0038] 3. Pot experiment to test herbicidal activity With reverse branch amaranth ( Amaranthus retroflexus For example, seeds of *Amaranthus retroflexus* were evenly sown in plastic pots (9×9cm) filled with sterilized potting soil (peat:vermiculite:sand = 3:1:1). At the two-true-leaf stage, the plants were sprayed with different concentrations of xanthocyanin solution (50 µg / mL, 100 µg / mL, 200 µg / mL, 500 µg / mL, and 1000 µg / mL) and a negative control (0 µg / mL), with three biological replicates for each concentration. The plants were grown under natural light in a greenhouse. After 21 days, all plants were harvested, and root length and seedling length were measured to obtain the desired results. Figure 7 The results are shown.

[0039] alfalfa ( Medicago sativa Taking (e.g.) as an example, the same pot experiment method as described above for *Amaranthus retroflexus* was used to cultivate the plants, and the results were as follows: Figure 6 The results are shown.

[0040] Depend on Figure 6 and Figure 7It was found that xanthocyanin had a dose-dependent herbicidal effect on amaranth and alfalfa seedlings. When the xanthocyanin concentration was 1000 μg / ml, amaranth showed significant leaf margin scorching and chlorotic white spots on the leaves. When the xanthocyanin concentration was 1000 μg / ml, alfalfa seedlings showed overall wilting and lodging.

[0041] 4. Herbicidal activity experiments at the molecular level PyMOL 2.6.0 software was used for analysis and plotting, and Discovery Studio 2021 Client was used to display the receptor-ligand interaction forces. Simulation analysis was performed on the xanthoside obtained in step 1 to study the binding ability of xanthoside to the 1SEZ protein of Arabidopsis thaliana protoporphyrinogen IX oxidase (PPO), obtaining results as follows: Figure 8 and Figure 9 The results are shown.

[0042] Protoporphyrinogen IX oxidase (PPO) is a key enzyme in the chlorophyll biosynthesis pathway, catalyzing the dehydrogenation of protoporphyrinogen IX to generate protoporphyrin IX—the seventh step in the chlorophyll synthesis pathway. Protoporphyrin IX is a direct precursor to chlorophyll, a crucial pigment involved in photosynthesis and essential for energy metabolism within chloroplasts. In photosynthetic organisms, inhibiting PPO enzyme activity blocks the formation of protoporphyrin IX, thereby interfering with normal chlorophyll synthesis. Simultaneously, the accumulation of the substrate protoporphyrinogen IX under light can spontaneously oxidize to produce reactive oxygen species, leading to cell membrane damage and plant death. This mechanism has been utilized by various commercial herbicides (such as flufenacet and ethoxyflufenazate).

[0043] Depend on Figure 8 and Figure 9 It was found that xanthoside can stably bind to the 1SEZ protein of PPO enzyme with a binding energy of -6.4 kcal / mol, exhibiting strong affinity. Docking mode analysis showed that xanthoside mainly forms hydrophobic interactions with Leu356, Phe392, Leu372, and Gly175 in the 1SEZ protein, with Phe392 exhibiting π-π stacking interactions with the aromatic ring of xanthoside. These interactions can interfere with the normal structure and function of the 1SEZ protein, thereby inhibiting PPO enzyme synthesis, affecting normal plant metabolism, and ultimately exerting herbicidal effects. This molecular docking simulation result indicates that xanthoside may block chlorophyll biosynthesis by binding to and inhibiting the activity of PPO enzyme. This reveals the potential molecular mechanism of action of xanthoside as a novel PPO inhibitor herbicide, providing an important theoretical basis for its further development into a highly efficient and environmentally friendly herbicide.

[0044] In summary, this invention provides an application of zineb in the preparation of plant-based herbicides and a plant-based herbicide, which has at least the following advantages: By applying xanthocyanin as the active ingredient in plant-based herbicides, effective suppression of various common farmland weeds was achieved. Experimental data showed that at concentrations ≥20 μg / mL, xanthocyanin exhibited detectable inhibitory effects on the root length and seedling height of Amaranthus retroflexus, Setaria viridis, alfalfa, and Chenopodium album, with the inhibitory effect significantly increasing with increasing concentration. In particular, a strong herbicidal effect was observed at a concentration of 1000 μg / mL, indicating that this concentration range possesses practical application feasibility and dose-dependent controllability.

[0045] Furthermore, the xanthocyanin provided by this invention is derived from natural plant secondary metabolites and belongs to the category of biodegradable phenolic compounds. It does not easily accumulate in the environment and does not cause soil degradation or water pollution, thus exhibiting higher ecological safety compared to chemically synthesized herbicides. Its mechanism of action is achieved by inhibiting key morphological development indicators during weed growth, including reducing seedling and root length, inducing leaf scorch and chlorotic white spots, and causing overall plant wilting and lodging, thereby effectively blocking the normal growth cycle of weeds.

[0046] Furthermore, this xanthocyanin can be obtained from Artemisia argyi via ethanol extraction, vacuum evaporation, and chromatographic separation. The preparation method is stable and controllable, and the abundant plant resources make it suitable for large-scale extraction and formulation development. As a herbicide, xanthocyanin can be formulated into different concentrations for pre-emergence or seedling stage application, either alone or in combination with other pesticides. This technical solution not only expands the application of xanthocyanin in agriculture but also provides new candidate substances and technical pathways for developing efficient, broad-spectrum, and low-residue bio-based herbicides, meeting the development needs of green agriculture, organic agriculture, and sustainable weed control.

[0047] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The application of a type of xanthocyanin in the preparation of plant-based herbicides, characterized in that, The concentration of the xanthocyanin in the herbicide product is ≥20 μg / mL.

2. The application according to claim 1, characterized in that, The concentration of the xanthocyanin in the herbicide product is 20~1000 μg / mL.

3. The application according to claim 1, characterized in that, The concentration of the xanthocyanin in the herbicide product is 100~1000 μg / mL.

4. The application according to claim 1, characterized in that, The xanthocyanin achieves its weed-control effect by reducing the seedling length of weeds, reducing the root length of weeds, causing the leaves to become scorched, causing the leaves to develop chlorotic white spots, or causing the weeds to fall over.

5. The application according to claim 1, characterized in that, The plant-based herbicide targets weeds including Amaranthus retroflexus (…). Amaranthus retroflexus ), foxtail grass ( Setaria viridis ), alfalfa Medicago sativa ) and gray-green quinoa ( Oxybasis glauca At least one of the following.

6. The application according to claim 1, characterized in that, The zanthoxylin is an active substance extracted from Artemisia argyi in Yili and has the following structure: .

7. The application according to claim 6, characterized in that, The method for preparing the zanthoxylin includes drying and pulverizing Artemisia argyi to obtain Artemisia argyi powder; extracting the Artemisia argyi powder and evaporating under reduced pressure to obtain Artemisia argyi extract; and obtaining the zanthoxylin by gradient elution and chromatographic separation of the Artemisia argyi extract.

8. The application according to claim 7, characterized in that, The solvent used for extracting the dried Artemisia argyi powder is 95% ethanol; And / or, the reduced pressure evaporation temperature is 40~50℃.

9. The application according to claim 7, characterized in that, The solvents used for gradient elution include petroleum ether and ethyl acetate; And / or, chromatographic separation includes separation using column chromatography and / or high-performance liquid chromatography.

10. A plant-based herbicide, characterized in that, The active substance includes xanthocyanin, and the concentration of xanthocyanin in the plant herbicide product is ≥20 μg / mL, preferably 20~1000 μg / mL, more preferably 100~1000 μg / mL.

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