Use of extract of valeriana traditional chinese medicinal materials as biostimulant

Abstract

The use of a valerian extract in plant growth. The agricultural field is lacking in terms of diversified usage methods for valerian and extracts thereof. Provided is a new use of a valerian extract. Specifically, the valerian extract is used for promoting plant growth.

Description

Application of valerian extracts from traditional Chinese medicine as biostimulants Technical Field

[0001] The invention belongs to the technical field of agriculture, and particularly relates to application of a valerian extract in plant growth. Background Art

[0002] Valerian is a plant of the family Valerianaceae. It is native to Asia, Europe, and North America, and is widely distributed in the northern temperate zones of the Americas, Europe, and Asia. In China, valerian is found in northeastern, northwestern, and southwestern provinces, growing in forests, shrublands, and alpine meadows at altitudes between 2,600 and 3,800 meters. Valerian prefers cool, humid climates and is cold-tolerant. It thrives in neutral or slightly alkaline sandy loam.

[0003] Valerian is commonly used as a traditional Chinese medicine. Its pungent, bitter, and warm properties enter the heart and liver meridians. The Compendium of Materia Medica states that it "calms the mind, regulates qi, and relieves pain." This indicates that it has calming effects, dispelling rheumatism, promoting the flow of qi and blood, and relieving pain. It is used to treat restlessness caused by internal depression, palpitations and insomnia, rheumatic pain, abdominal distension and pain, dysmenorrhea, amenorrhea, and traumatic injuries. Valerian, commonly used as a medicinal herb, is selected from the roots and rhizomes of Valeriana officinalis L. or Valeriana officinalis Var. Latifolia Miq., both of the Patrinaceae family.

[0004] Modern research indicates that valerian extract has the following main functions: ① antidepressant; ② sedative, hypnotic, and anticonvulsant effects; and ③ antibacterial, antiviral, and antitumor effects. Experimental studies have shown that it can enhance the inhibitory process of the cerebral cortex, reduce reflex excitability, and relieve smooth muscle spasms, resulting in sedative, hypnotic, antispasmodic, and analgesic effects.

[0005] According to current research, both domestically and internationally, the main active ingredients of valerian are found in its volatile oils and non-volatile esters and organic acids. The main active ingredients in the volatile oils are oxygenated derivatives of terpenes, such as borneol, valeranone, caryophyllene, borneol, caryophyllene, and limonene. The main active ingredients in the non-volatile substances are iridoid esters and terpene acids, including valerenyl triesters, isovalerenyl triesters, and hydrogenated valerenyl triesters, and valerenic acid, acetylvailic acid, and valerenal. These substances all have antispasmodic and analgesic effects on gastrointestinal smooth muscle, as well as sedative and tranquilizing effects. The three types of substances also exhibit synergistic effects. The quality standards for valerian and its preparations in various pharmacopoeias also use the content of certain substances as criteria. For example, the United States uses total valerenic acid, Japan uses volatile oils, and Germany uses both iridoid esters and volatile oils.

[0006] Summary of the Invention

[0007] The present invention is the first to discover that valerian extract has the effect of promoting plant growth, which expands the application range of valerian extract and also provides a new option for regulating plant growth in agriculture.

[0008] Experiments have found that the present invention uses different solvents and methods to extract various medicinal materials and then conduct activity tests. The results show that valerian volatile oil can significantly promote the growth of various crops such as corn, wheat, cabbage, cucumber, rice, etc., including their plant height, stem diameter, leaf photosynthetic rate, root length and other indicators.

[0009] The present invention provides an application of a valerian extract in promoting plant growth; or provides a method for promoting plant growth, which is achieved by applying the valerian extract to plants.

[0010] The valerian extract of the present invention is selected from the extract of the root or rhizome of Valerian (Valeriana officinalis L.) or Valerian (Valeriana officinalis Var. Latifolia Miq) of the Patrinaceae family.

[0011] In the present invention, promoting plant growth includes promoting germination, or promoting the growth of one or more of roots, stems, leaves, or flowers and fruits. For example, promoting the growth of root length, stem diameter, plant height, leaf width, leaf length, leaf number, leaf area, biomass, chlorophyll content, yield, etc.

[0012] In the present invention, the valerian extract is used as an active ingredient of the growth promoter and can be used alone or in combination with other products.

[0013] The "biostimulants" mentioned in this invention include substances containing certain ingredients and microorganisms. When these substances are applied to the plant itself or its rhizosphere, they have a positive stimulating effect on the natural processes of the plant, including enhancing or benefiting the plant's physiological functions, nutrient absorption, resistance to abiotic stress and improvement of crop quality, but are not related to nutritional content.

[0014] The "growth promoter" mentioned in the present invention refers to a plant growth promoter, which is a biologically active substance that can promote cell division, differentiation and elongation growth, or promote the growth of plant vegetative organs and the development of reproductive organs.

[0015] The present invention finds that valerian extract is a natural plant growth promoter.

[0016] In the present invention, the extract is valerian volatile oil, or an extract containing valerian volatile oil.

[0017] Volatile oil, also known as essential oil, is a general term for a class of volatile oily components that are immiscible with water and can be obtained by steam distillation. They have an aromatic smell.

[0018] Volatility refers to the ability of a liquid substance to transform into a gaseous state at a temperature below its boiling point, as well as the ability of some gaseous solutes to escape from the solution.

[0019] The extract containing valerian volatile oil includes but is not limited to an organic solvent extract containing valerian volatile oil, a supercritical carbon dioxide extract containing valerian volatile oil, and a steam distillation extract containing valerian volatile oil.

[0020] The volatile oil content in valerian is usually 0.5% to 2%, and the main components of the volatile oil in valerian are basically divided into monoterpenes and sesquiterpenes.

[0021] The valerian volatile oil used in the present invention can be obtained by purchasing commercial products, or can be prepared by conventional methods or existing technologies.

[0022] The commonly used extraction methods for volatile oils include steam distillation, extraction, adsorption, etc. (see "Plant Natural Product Extraction Technology", Chemical Industry Press, 2022.7, pages 100-110).

[0023] Steam distillation: Volatile oils and water do not mix. However, when heated, the combined vapor pressure of the volatile oil and water equals atmospheric pressure, causing the solution to boil. The volatile oil and water vapor are then distilled together, and the volatile oil is then separated based on the principle that the two do not mix. The process involves heating the plant material and water mixture, generating water vapor, condensing the water vapor, and collecting the condensate. As the condensate separates, the volatile oil and water automatically separate. The oil layer is collected, yielding the volatile oil. If the entire condensate is collected, a steam distillation extract containing the volatile oil is obtained.

[0024] Extraction method: Utilizing the ability of volatile oils to dissolve well in certain volatile solvents, volatile oils are extracted from plants and then the solvents are evaporated under reduced pressure to obtain an extract containing the volatile oils. Commonly used solvents include high-concentration ethanol (95% ethanol, anhydrous ethanol, etc.), petroleum ether, ether, n-hexane, etc.

[0025] Adsorption: This is mostly physical adsorption, caused by van der Waals forces. Commonly used adsorbents include silica gel and activated carbon. Adsorption is an exothermic process, meaning the entropy of the system decreases, lowering the interfacial free energy. It is an automatic process. Adsorption is mostly multi-molecular, and the adsorbent itself remains unchanged. Volatile oils can be recovered through desorption.

[0026] In addition, supercritical carbon dioxide extraction is now also commonly used to extract volatile oils. Supercritical carbon dioxide extraction refers to the process of contacting supercritical carbon dioxide with the substance to be separated under a supercritical state, so that it can selectively extract components with different polarity, boiling points and molecular weights in sequence. Of course, the extracts obtained corresponding to each pressure range cannot be single, but the conditions can be controlled to obtain the optimal proportion of mixed components. Then, by using the method of reducing pressure and increasing temperature, the supercritical fluid is turned into an ordinary gas, and the extracted substance is completely or basically precipitated, thereby achieving the purpose of separation and purification. Therefore, the supercritical fluid carbon dioxide extraction process is composed of a combination of extraction and separation.

[0027] Methods for extracting valerian essential oil have been documented in the prior art. For example, "Yang Jie. Valerian Essential Oil Extraction Process, Chemical Composition Analysis, and Active Function Research [D]. Central South University of Forestry and Technology, 2009" discloses steam distillation for extracting valerian essential oil. Chinese patent applications CN106281708A and CN111733016A also disclose methods for extracting valerian essential oil using supercritical carbon dioxide fluid extraction. Chinese patent application CN108998243A discloses microwave-assisted extraction using n-hexane.

[0028] In the present invention, the extract containing valerian volatile oil generally refers to an extract prepared by an extraction method. During solvent extraction, other components are extracted in addition to the volatile oil, and the volatile oil content is reduced, so the dosage needs to be increased when used.

[0029] The extraction method of the present invention can be carried out in the following ways: heating extraction, percolation extraction, ultrasonic extraction, microwave extraction, immersion extraction, supercritical extraction, etc.

[0030] After extraction, other conventional preparation steps are also included, such as filtration, concentration, centrifugation, drying, evaporation, etc.

[0031] In order to extract the biologically active ingredients in valerian as much as possible, valerian can be crushed and sieved.

[0032] In some specific embodiments of the present invention, during extraction, the liquid-to-solid ratio of the added solvent to valerian is 4 to 20:1 (v / v), for example, 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, etc.

[0033] In some specific embodiments of the present invention, during heating extraction, heating leaching or heating reflux can be used for extraction, and the extraction temperature is 40°C to 95°C, for example, it can be 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, 95°C, etc.

[0034] The number of extractions is selected according to demand, generally 1 to 4 times, with 2 or 3 times being the most appropriate.

[0035] In addition, the extraction conditions of conventional extraction methods such as ultrasonic extraction, microwave extraction, and supercritical extraction in the present invention can be obtained by conventional adjustments by those skilled in the art.

[0036] The plants described in the present invention include but are not limited to economic crops, food crops, such as small cruciferous vegetables, field crops, fruit trees, solanaceous plants, and other types of crops.

[0037] Nightshade vegetables refer to vegetables in the Solanaceae family whose edible organs are berries, such as tomatoes, peppers, and cucumbers.

[0038] Cruciferous vegetables can be divided into leafy vegetables (Chinese cabbage), stalk vegetables (red stalk, Guangdong choy sum), head vegetables (cabbage), flower vegetables (cauliflower, broccoli) and underground tuber vegetables (radish) according to their edible parts.

[0039] Fruit trees refer to trees with edible fruits, a general term for perennial plants and their rootstocks that can provide edible fruits and seeds.

[0040] Field crops include wheat, rice, sorghum, corn, cotton, forage, etc.

[0041] The so-called "economic crops" are of various types, including but not limited to fiber crops (such as cotton, hemp, etc.), oil crops (such as sesame, peanuts, etc.), sugar crops (such as sugarcane, sugar beets, etc.), hobby crops (tobacco), medicinal crops, dye crops, ornamental crops, fruits (such as citrus, kiwi, cherry, pear, apple, etc.) and other economic crops (such as lettuce, pepper, Chinese cabbage, lettuce, Shanghai green, etc.).

[0042] The “food crops” mentioned above include but are not limited to cereal crops (wheat, rice, corn), tuber crops (including sweet potatoes, potatoes, etc.) and legume crops (including soybeans, broad beans, peas, mung beans, etc.).

[0043] The present invention also provides a biostimulant or plant growth promoter, which uses valerian extract as an active ingredient.

[0044] In the product of the present invention, valerian extract or valerian extract solution can be directly used as a single dose. The "single dose" mentioned in the present invention refers to a product that uses valerian extract as the sole active ingredient and does not add other excipients.

[0045] In order to make the product more stable and convenient for transportation and storage, it can be made into an agricultural product, for example, by adding auxiliary materials to make a corresponding dosage form. The auxiliary materials can be conventional auxiliary materials in the field, such as dispersants, wetting agents, binders, emulsifiers, stabilizers, solvents, etc.

[0046] On the other hand, the valerian extract of the present invention can be used as a synergistic ingredient in combination with foliar fertilizers, water-soluble fertilizers, compound fertilizers, pesticides, or biostimulants.

[0047] In the present invention, the dosage forms of the agricultural products include but are not limited to emulsifiable concentrates, suspensions, wettable powders, dusts, granules, aqueous solutions, mother liquors, mother powders, microemulsions, microcapsule suspensions, aqueous emulsions, tablets, smoke agents, sustained-release agents, and seed coating agents.

[0048] Emulsifiable concentrates (ECs) are a type of pesticide formulation. They are formed by dissolving a high concentration of the active ingredient in a solvent and adding an emulsifier. They are typically diluted with a large amount of water to form a stable emulsion and then applied using a sprayer. Low-volume and even ultra-low-volume sprays are also possible. They can be used directly or diluted with water before spraying on crops.

[0049] Wettable powder is a very fine dry preparation obtained by mixing and crushing the original drug, filler, surfactant and other additives.

[0050] A suspension concentrate (SC) is a formulation in which a solid technical drug is uniformly dispersed in water as particles smaller than 4 microns. Its international designation is SC. It has a fine particle size, typically 0.1 to 3 μm, and a high suspension rate. Suspension concentrates are categorized as water-based suspension concentrates and oil-based suspension concentrates. Water-based suspension concentrates use water as the suspending medium, while oil-based suspension concentrates use oils as the suspending medium and do not contain water. Commonly used oils are vegetable oils, such as corn oil and rapeseed oil. Suspension concentrates eliminate the need for organic solvents and are an ideal formulation for processing solid technical drugs. A suspension concentrate is a mixture of solid powder and liquid suspended in water. It must be shaken well before use, diluted with water, and then sprayed on crops. Suspension concentrates are easy to carry and dilute, can be sprayed evenly on crops, and have good adhesion and long-lasting effectiveness.

[0051] Dust refers to the powder of the original drug, or a powder prepared by adding a certain diluent. It can be sprayed directly on crops with a simple duster. It is highly efficient, has low adhesion to crops, leaves little residue, and is less likely to cause drug damage.

[0052] Granules, also known as granules, are a solid dosage form obtained by mixing the active ingredient with a carrier, adhesive, dispersant, wetting agent, stabilizer, and other additives. Performance requirements primarily include fineness, uniformity, storage stability, hardness, and disintegration. Granules have the largest particle size among solid dosage forms, ranging from 300 to 1700 μm in diameter. They offer the advantages of ease of use, minimal diffusion, and long-lasting efficacy.

[0053] Aqueous agent is an aqueous solution of the original drug. The drug is evenly dispersed in water in the form of ions or molecules. The concentration of the drug depends on the water solubility of the original drug, which is generally its maximum solubility. It is then diluted with water when used.

[0054] Mother liquor or mother powder is a semi-finished product made from the original drug and needs to be further processed into the final product.

[0055] In the present invention, when used, the single agent or agricultural product prepared from the valerian extract is used to treat seeds, spray on leaves or apply to roots.

[0056] In the present invention, when the valerian extract or its product is prepared into a solution for promoting seed germination or plant growth, the concentration of the valerian extract in the solution can be selected according to actual needs.

[0057] Including but not limited to 0.01~5000ppm, can be selected from 500~3000ppm, 1000~2000ppm, specifically such as 1000±100ppm......1500±100ppm......2000±100ppm......2500±100ppm and the like, for example selected from 0.01ppm,......0.02ppm,......0.03ppm,......0.1ppm,......0.2ppm,......0.3ppm,......1.0ppm,......2.0ppm,......3.0ppm,......10ppm,......20ppm,......30ppm and the like.

[0058] The seeds are treated by mixing a single agent or an agricultural product made from the valerian extract with the seeds, so that the valerian extract adheres to or is absorbed by the surface of the seeds. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 is a gas chromatogram of valerian essential oil;

[0060] Figure 2 is the gas phase detection spectrum of valerian ethanol extract. DETAILED DESCRIPTION

[0061] The technical scheme of the present invention will be clearly and completely described below. Of course, the described embodiments are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention. It should be noted that if there is a process that is not particularly described in detail below, it is all that those skilled in the art can realize or understand with reference to the prior art. Reagents or instruments used that do not indicate the manufacturer are deemed to be conventional products that can be purchased commercially.

[0062] In the embodiments of the present invention, the valerian used is sourced from the Bozhou Chinese medicinal materials market in Anhui Province.

[0063] The valerian volatile oil of the present invention can be obtained by purchasing commercial products or prepared by conventional extraction methods of volatile oils. The extraction methods of volatile oils in the following examples are merely reference to common knowledge to prepare the raw materials used in the experiments and should not be construed as limiting the methods for obtaining valerian volatile oils.

[0064] Example 1

[0065] Valerian herbs are taken and crushed, soaked and moistened in a steam distillation device, and reflux extracted until no more volatile oil is extracted, and the volatile oil is naturally cooled to room temperature after stopping heating, and the volatile oil is collected from the volatile oil extraction device to obtain valerian volatile oil.

[0066] Example 2

[0067] Take an appropriate amount of valerian crude powder, use one solvent selected from water, ethanol (anhydrous ethanol), ethyl acetate, and petroleum ether at a solid-liquid ratio of 1:8, perform ultrasonic extraction for 1 hour, extract twice, combine the two extracts, and concentrate to dryness under reduced pressure to obtain a valerian extract.

[0068] The components of the valerian extract were fully scanned using gas chromatography and compared with existing databases. The specific conditions were: a chromatographic column DB-1 capillary column (100% dimethylpolysiloxane as the stationary phase) (30m×0.25mm, 0.25μm) was selected; the column temperature was 100°C, the injection port temperature was 230°C, the detector (FID) temperature was 250°C; and the split ratio was 10:1. Figure 1 is a gas phase detection spectrum of valerian volatile oil. When the volatile oil of the present invention is applied to plants, it is prepared with sterile water, and an appropriate amount of ethanol (such as 5%) and a surfactant (such as 5% castor oil polyoxyethylene ether) are added to ensure that the Cnidium monnieri volatile oil is evenly dispersed in the water.

[0069] Figure 2 is a gas phase detection spectrum of the ethanol extract of valerian. When the ethanol extract of the present invention is applied to plants, it can be directly diluted with sterile water.

[0070] Example 3

[0071] Through the indoor plant growth promotion test, the specific test process is: the wheat seedlings / corn seedlings / Chinese cabbage seedlings that have been cultivated to a plant height of 5-10 cm and have the same growth are tested by leaf spraying. The leaves are sprayed according to the standard of moistening each leaf. After the treatment is completed, they are placed in the culture room. The temperature of the culture room is 22℃~25℃ and the humidity is 60%~68%. The Chinese cabbage is medicated twice throughout the whole process, with an interval of 7 days each time. The corn and wheat are medicated once throughout the whole process, and at least 20 plants are treated for each treatment. The positive control treatment is sprayed with 5000 times the recommended concentration of seaweed extract (10% powder, Haijingling, provided by the manufacturer in April 2022), and the blank control is sprayed with an equal amount of water. After application, each treatment group is cultured under the same natural conditions.

[0072] Survey Method: Before the experiment, the aboveground height and SPAD value of each seedling were measured. One week after the second application, the aboveground height and the SPAD value of the same leaf were measured. SPAD values ​​were measured three times in a row, and the average value was recorded. Root length, plant fresh weight, and root fresh weight were also measured. The growth rates and relative growth rates of plant height, SPAD values, root length, plant fresh weight, and root fresh weight before and after the treatment were calculated. (Note: Since the root length, plant fresh weight and root fresh weight could not be measured before drug application, all conditions of the seedlings were kept consistent from the beginning of germination in the early stage of the experiment. The test materials with growth vigor of -2 and uniform size were selected. It can be regarded that all treatment indicators in the early stage were consistent, and only the growth rate after drug application was calculated.) Plant height growth rate = (plant height after treatment - plant height before treatment) * 100% plant height before treatment; Relative growth rate = (plant height growth rate after treatment - plant height growth rate of blank) * 100% / plant height growth rate of blank; SPAD change value = SPAD after treatment - SPAD before treatment; Root length growth rate = (root length after treatment - root length of blank) * 100% / root length of blank; Plant fresh weight growth rate = (plant fresh weight of treatment - plant fresh weight of blank) * 100% / plant fresh weight of blank; Root fresh weight growth rate = (root fresh weight of treatment - root fresh weight of blank) * 100% / root fresh weight of blank.

[0073] The experimental treatment settings were as shown in Table 1 for foliar application. The results of the index measurements of each treatment group 7-14 days after application are shown in Tables 2-7 below. The data in the tables are all average values.

[0074] Table 1 Experimental treatment settings

[0075] Table 2 Results of index determination of each treatment group before and after treatment of pakchoy

[0076] Table 3 Results of index determination of each treatment group before and after treatment of pakchoy

[0077] Table 4 Results of the indicators of each treatment group before and after the application of corn

[0078] Table 5 Results of the indicators of each treatment group before and after the application of the corn

[0079] Table 6 Wheat index test results of each treatment group before and after drug application

[0080] Table 7 Wheat index test results of each treatment group before and after drug application

Claims

1. A method for promoting plant growth, characterized in that: Apply valerian extract to the plant.

2. The method according to claim 1, characterized in that The promoting plant growth includes promoting seed germination, and / or promoting the growth of one or more of roots, stems, leaves, or flowers and fruits.

3. The method according to claim 1, characterized in that: The valerian extract is valerian volatile oil.

4. The method according to claim 1, characterized in that: The valerian extract is an extract containing valerian volatile oil.

5. The method according to claim 4, characterized in that The extract containing valerian volatile oil includes but is not limited to ethanol, petroleum ether, n-hexane or ether extract; wherein the valerian ethanol extract is extracted using ethanol with a concentration of more than 95%.

6. The method according to claim 5, characterized in that The valerian ethanol extract is extracted using anhydrous ethanol.

7. The method according to claim 1, characterized in that Plants whose growth is promoted include cruciferous vegetables, field crops, fruit trees, and nightshades.

8. The method according to claim 1, characterized in that When in use, the valerian extract is made into an agricultural product, and the agricultural product also includes auxiliary materials, which are one or more of a dispersant, a wetting agent, a binder, an emulsifier, a stabilizer, and a solvent.

9. The method according to claim 8, wherein the dosage form of the product is emulsifiable concentrate, suspension, wettable powder, powder, granule, aqueous solution, mother liquor, mother powder, microemulsion, microcapsule suspension, aqueous emulsion, tablet, smoke agent, sustained release agent, seed coating agent.

10. The method according to claim 8, characterized in that The agricultural product made from the valerian extract is used in combination with foliar fertilizer, water-soluble fertilizer, compound fertilizer, pesticide, or biostimulant.

11. The method according to claim 8, characterized in that When using, treat seeds, spray leaves or irrigate roots with a single dose of valerian extract or the prepared agricultural products.

12. A biostimulant or growth promoter, characterized in that: It contains valerian extract as its active ingredient.

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