Use of ester compound and related extract as biostimulant

Abstract

A use of bornyl acetate or a salt thereof and a plant extract containing bornyl acetate in plant growth. In the agricultural field, the use of bornyl acetate or the salt thereof and the plant extract containing bornyl acetate is relatively limited. Provided is a novel use of bornyl acetate or the salt thereof and the plant extract containing bornyl acetate, specifically, the use in promoting plant growth.

Description

Application of ester compounds and related extracts as biostimulants Technical Field

[0001] The invention belongs to the technical field of agriculture, and particularly relates to an ester compound and the application of multiple extracts containing the ester compound as biostimulants. Background Art

[0002] Bornyl acetate, also known as borneol acetate, occurs naturally in fir oil and some pine essential oils, usually as the levorotatory isomer. It can be isolated from essential oils or produced by acetylation of borneol with acetic anhydride.

[0003] Bornyl acetate has the following uses: ① As a fragrance. It is used in cosmetics, soaps, indoor disinfectants, air fresheners, room spray fragrances, bath fragrances, and body powders; it is often used to formulate fragrances such as fougere, lavender, cologne, chypre, and forest-flavored fragrances. ② As a food flavoring. It can be used to formulate mint, pineapple, and spice-flavored fragrances. ③ Bornyl acetate can also be used as a solvent and plasticizer for cellulose nitrocellulose.

[0004] Summary of the Invention

[0005] The present invention discovers for the first time that bornyl acetate has the effect of promoting plant growth, expands the application range of bornyl acetate, and also provides a new option for regulating plant growth in agriculture.

[0006] Experiments have found that bornyl acetate can significantly promote the growth of various crops, including corn, wheat, cabbage, cucumber, and rice, including various indicators such as plant height, stem diameter, leaf photosynthetic rate, and root length. The present invention provides the use of bornyl acetate, its salts, or isomers, or plant extracts containing bornyl acetate, or its salts or isomers, in the preparation of biostimulants.

[0007] Alternatively, a method for promoting plant growth is provided, which is achieved by applying bornyl acetate to the plants.

[0008] Growth promotion can promote cell division, differentiation, and elongation, or promote the growth of plant vegetative organs and the development of reproductive organs. 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, it can promote the growth of root length, stem diameter, plant height, leaf width, leaf length, leaf number, leaf area, biomass, chlorophyll content, yield, etc.

[0009] The present invention also found that the extracts of valerian, cnidium monnieri and amomum villosum also have the effect of promoting plant growth. Further detection by chromatography found that these extracts all contain bornyl acetate.

[0010] Based on this discovery, the present invention also provides the use of a plant extract containing bornyl acetate or its salts and isomers in promoting plant growth.

[0011] The plant extract containing bornyl acetate or its salt is selected from extracts of valerian, cnidium monnieri, amomum villosum, fir fruit, wormwood leaf, cassia twig, forsythia, houttuynia cordata and the like. Bornyl acetate is also known to exist in essential oils of other pine plants.

[0012] The present invention has found that plant extracts containing bornyl acetate at concentrations ranging from 1 mg / ml to several hundred mg / ml exhibit plant growth-promoting activity. Based on this, plant extracts containing bornyl acetate at or above 1 mg / ml can be selected for the present invention, further selected from 1 to 1000 mg / ml, and even further selected from 10 to 500 mg / ml. The above ranges are merely examples and should not be construed as limiting the content to the aforementioned ranges.

[0013] Based on the fact that the experiments of the present invention have proved that bornyl acetate has the above-mentioned activity, combined with the similar activities of Cnidium monnieri extract, Amomum villosum extract, Houttuynia cordata extract, and Valerian extract containing bornyl acetate, it can be reasonably expected that any plant extract containing bornyl acetate can produce the effect of promoting plant growth.

[0014] Valerian is a plant of the family Valerianaaceae. It is commonly used as a traditional Chinese medicine. The 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 which are also Valerianaaceae plants.

[0015] Cnidium monnieri (L.) Cuss. is the dried mature fruit of the Apiaceae plant, Cnidium monnieri (L.) Cuss. It is distributed in East China, Central South, Southwest China, Northwest China, North China, and Northeast China.

[0016] Amomum villosum is the dried mature fruit of Amomum villosum Lour., Amomum villosum Lour.var.xanthioides TLWu et Senjen or Amomum longiligulare TLWu, all of the Zingiberaceae family.

[0017] Houttuynia cordata is derived from the dried aerial parts of Houttuynia cordata Thunb., a plant of the Saururaceae family.

[0018] In a specific embodiment of the present invention, it was detected that the various extracts mentioned above contained different amounts of bornyl acetate.

[0019] In addition, the content of bornyl acetate in Siberian pine needle oil is as high as 30% to 40%, the content in Houttuynia cordata volatile oil is about 10 to 30 mg / mL, the content in Amomum villosum volatile oil is about 300 to 400 mg / mL, the content in valerian volatile oil is about 10 to 30 mg / mL, the content in Cnidium monnieri volatile oil is about 90 to 130 mg / mL, the content in Artemisia argyi volatile oil is about 0.2 to 0.5 mg / mL, and the content in Forsythia suspensa volatile oil is about 0.3 to 0.6 mg / mL.

[0020] In one embodiment of the present invention, the detection method of the present invention is used, and the contents of bornyl acetate in the following volatile oils are respectively:

[0021] In addition to the above plants, other plants also contain bornyl acetate: Stephania tetrandra, frankincense, ginger, agrimony, perilla, tonka bean, loquat leaf, cinnamon, mulberry bark, Acorus calamus, magnolia, polygonatum, turmeric, coriander, trumpet creeper, chrysanthemum, honeysuckle, Magnolia officinalis, Salvia miltiorrhiza, Schisandra chinensis, Morinda officinalis, Asarum, Evodia rutaecarpa, Rhodiola rosea, Buddha's hand, Curcuma zedoaria, etc.

[0022] The isomers include stereoisomers, optical isomers and the like.

[0023] Bornyl acetate has two optical isomers: the l-isomer and the d-isomer. The l-isomer is found in conifer oils of conifers, such as Canadian hemlock, white fir, white spruce, red spruce, and eastern spruce, as well as in coriander, thyme, and chemicalbook valerian essential oils. The d-isomer is primarily found in conifer oils of pines, such as white frost pine and black pine, as well as in lavender essential oil and cypress leaves.

[0024] Isoborneol acetate is a common isomer of bornyl acetate.

[0025] In the present invention, bornyl acetate or its salts or isomers, or plant extracts containing bornyl acetate or its salts or isomers are used as active ingredients of the growth promoter and can be used alone or in combination with other products.

[0026] For example, it may include but is not limited to the following combinations:

[0027] (1) one of Cnidium monnieri extract, Amomum villosum extract, Houttuynia cordata extract, and Valerian extract, used in combination with bornyl acetate;

[0028] (2) The Cnidium monnieri extract and Amomum villosum extract are used in combination;

[0029] (3) The Cnidium monnieri extract is used in combination with Houttuynia cordata extract;

[0030] (4) The Cnidium monnieri extract is used in combination with the Valerian extract;

[0031] (5) Amomum villosum extract and Houttuynia cordata extract are used in combination;

[0032] (6) Amomum villosum extract and valerian extract are used in combination;

[0033] (7) Houttuynia cordata extract and valerian extract are used in combination;

[0034] (8) Use Cnidium monnieri extract, Amomum villosum extract, and Houttuynia cordata extract in combination;

[0035] (9) Combination use of Cnidium monnieri extract, Houttuynia cordata extract, and Valerian extract;

[0036] (10) Cnidium monnieri extract, Amomum villosum extract, and Valerian extract are used in combination;

[0037] (11) Amomum villosum extract, Houttuynia cordata extract, and Valerian extract are used in combination;

[0038] (12) Combination use of Cnidium monnieri extract, Amomum villosum extract, Houttuynia cordata extract, and Valerian extract; etc.

[0039] 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.

[0040] 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.

[0041] The present invention finds that bornyl acetate or its salt, or a plant extract containing bornyl acetate or its salt is a natural plant growth promoter.

[0042] In the present invention, the extract is an extract of water and / or an organic solvent.

[0043] Bornyl acetate is slightly soluble in water and easily soluble in organic solvents. In the present invention, the organic solvent includes one or more of alcohol solvents, ester solvents, ketone solvents, and petroleum ether.

[0044] "Alcohols" are compounds in which hydrogen atoms in the side chains of aliphatic, alicyclic, or aromatic hydrocarbons are replaced by hydroxyl groups. Alcohols are widely used as solvents and are inexpensive. The "alcohol solvents" described herein generally use alcohols with less than 12 carbon atoms, including but not limited to ethanol or hydrous ethanol, methanol or hydrous methanol, and n-butanol or hydrous n-butanol.

[0045] "Ester" refers to a class of organic compounds generated by the reaction of an acid (carboxylic acid or inorganic oxygen-containing acid) with an alcohol. The "ester solvent" described in the present invention is a lower ester in a liquid state, including but not limited to ethyl acetate and methyl acetate.

[0046] "Ketone" is a compound in which a carbonyl group is connected to two hydrocarbon groups. The "ketone solvent" of the present invention is a liquid lower ketone, including but not limited to acetone.

[0047] In the present invention, the ethanol content in the aqueous ethanol is selected from 5 to 95% v / v, further selected from 10 to 95% v / v, for example, it can be selected from 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% v / v and other concentrations of ethanol.

[0048] The methanol content in the aqueous methanol is selected from 5 to 95% v / v, further selected from 10 to 95% v / v. For example, the methanol concentration may be selected from 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% v / v.

[0049] The extraction method of the present invention can adopt conventional plant extraction methods, such as but not limited to heating extraction, percolation extraction, ultrasonic extraction, microwave extraction, immersion extraction, supercritical extraction, etc.

[0050] After extraction, other conventional preparation steps are also included, such as filtration, concentration, centrifugation, drying, evaporation, etc.

[0051] 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.

[0052] 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.

[0053] The extract of the present invention may also be a volatile oil or an extract containing volatile oil, such as Cnidium monnieri volatile oil, an organic solvent extract containing Cnidium monnieri volatile oil, a supercritical carbon dioxide extract containing Cnidium monnieri volatile oil, a steam distillation extract containing Cnidium monnieri volatile oil, valerian volatile oil, an organic solvent extract containing valerian volatile oil, a supercritical carbon dioxide extract containing valerian volatile oil, a steam distillation extract containing valerian volatile oil, Amomum villosum volatile oil, an organic solvent extract containing Amomum villosum volatile oil, a supercritical carbon dioxide extract containing Amomum villosum volatile oil, a steam distillation extract containing Amomum villosum volatile oil, Houttuynia cordata volatile oil, an organic solvent extract containing Houttuynia cordata volatile oil, a supercritical carbon dioxide extract containing Houttuynia cordata volatile oil, a steam distillation extract containing Houttuynia cordata volatile oil, and the like.

[0054] 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).

[0055] 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.

[0056] 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.

[0057] 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.

[0058] 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.

[0059] In order to extract as much biologically active ingredients from plants as possible, the plants can be crushed and sieved.

[0060] In some specific embodiments of the present invention, during extraction, the ratio of the added solvent to the plant liquid is 6 to 10:1 (v / v), for example, it can be 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, etc.

[0061] 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.

[0062] The number of extractions is selected according to demand, generally 1 to 4 times, with 2 or 3 times being the most appropriate.

[0063] 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.

[0064] The extraction methods of the above-mentioned volatile oils are reported in the prior art, for example:

[0065] "Zhu Huayu, Song Xingliang, Sun Aide. GC-MS analysis of components extracted from Cnidium monnieri by supercritical CO2 extraction [J]. Journal of Linyi Normal University, 2006(06):49-55." This paper reports that the effective components of Cnidium monnieri were extracted by supercritical CO2 extraction with ether as a modifier, and the chemical components and relative contents were analyzed by GC / MS. The results showed that the yield of supercritical CO2 extraction was higher than that of steam distillation, with a total extraction rate of 8.46%. "Zhu Ying, Gu Yaohua, Zhu Lei. GC-MS analysis of volatile oil components in Cnidium monnieri from two production areas [J]. Chinese Pharmacy, 2008(33):2603-2605." This paper reports that the volatile oil of Cnidium monnieri was extracted by steam distillation.

[0066] "Yang Jie. Valerian Essential Oil Extraction Process, Chemical Composition Analysis, and Active Function Research [D]. Central South University of Forestry and Technology, 2009." discloses the extraction of valerian essential oil using steam distillation. 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.

[0067] "Huang Lu, Fu Xingqing, Xiao Jia, et al. GC-MS analysis of volatile components from different parts of Amomum villosum [J]. Journal of Chengdu University of Traditional Chinese Medicine, 2020, 43(02): 42-46.", discloses the extraction of Amomum villosum volatile oil by steam distillation. "Lin Jingming, Zheng Yuhua, Chen Feilong, et al. Analysis of components of Amomum villosum volatile oil extracted by supercritical CO2 fluid [J]. Chinese Medicinal Materials, 2000, 23(01): 37-39.", discloses the extraction of Amomum villosum volatile oil by supercritical extraction.

[0068] "Zhang Chengcheng, Yang Lingyu, Geerile, et al. Process optimization of supercritical CO2 extraction of Houttuynia cordata volatile oil [J]. Jiangxi Traditional Chinese Medicine, 2017, 48(06): 63-65.", published "Study on the optimal process conditions for supercritical CO2 extraction of Houttuynia cordata volatile oil components. Methods: Taking the volatile oil extraction rate as an indicator, an orthogonal experimental design was carried out on the basis of single factor investigation of extraction pressure, temperature and time to determine the optimal process conditions. Results: The optimal extraction conditions were pressure 30 MPa, temperature 45℃, time 100 min, separation I pressure 7 MPa; temperature 50℃, CO2 flow rate controlled at 45 kg / h, and the volatile oil yield was 1.98%." "Zhang Zhuangli, Zhao Ning, Zhao Zhihong, et al. Optimization of extraction process of Houttuynia cordata volatile oil [J]. Chinese Journal of Medical Science, 2015, 5(01): 82-85." "The optimal process conditions for extracting Houttuynia cordata volatile oil by steam distillation were studied. Methods The volatile oil extraction rate was used as the evaluation index. Single factor investigation and orthogonal experimental design were used to investigate the effects of crushing degree, soaking time, material-liquid ratio and reflux time on the volatile oil extraction rate. Results The optimal process conditions for extracting Houttuynia cordata volatile oil by steam distillation were as follows: crushing the herbal medicine to coarse powder, material-liquid ratio of 1:10 (g / g), soaking for 10 hours, and heating and refluxing for 10 hours."

[0069] 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.

[0070] Nightshade vegetables refer to vegetables in the Solanaceae family whose edible organs are berries, such as tomatoes, peppers, and cucumbers.

[0071] 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.

[0072] Fruit trees refer to trees with edible fruits, a general term for perennial plants and their rootstocks that can provide edible fruits and seeds.

[0073] Field crops include wheat, rice, sorghum, corn, cotton, forage, etc.

[0074] 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.).

[0075] 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.).

[0076] The present invention also provides a biostimulant or plant growth promoter, which uses bornyl acetate or its salt, or a plant extract containing bornyl acetate or its salt as an active ingredient.

[0077] As mentioned above, the plant extract includes one or a mixture of multiple of Cnidium monnieri extract, Amomum villosum extract and Houttuynia cordata extract.

[0078] In the product of the present invention, bornyl acetate or its salt, or a plant extract containing bornyl acetate or its salt can be directly used as a single dose. The "single dose" mentioned in the present invention refers to a product that uses bornyl acetate or its salt, or a plant extract containing bornyl acetate or its salt as the sole active ingredient, and does not add other excipients.

[0079] 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.

[0080] On the other hand, the bornyl acetate or its salt, or the plant extract containing bornyl acetate or its salt described in the present invention can be used as a synergistic ingredient in combination with products such as foliar fertilizers, water-soluble fertilizers, compound fertilizers, pesticides, or biostimulants.

[0081] In the present invention, the dosage form of the agricultural product includes but is not limited to emulsifiable concentrate, suspension concentrate, wettable powder, dust, granule, aqueous solution, mother liquor or mother powder.

[0082] 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.

[0083] Wettable powder is a very fine dry preparation obtained by mixing and crushing the original drug, filler, surfactant and other additives.

[0084] 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.

[0085] 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.

[0086] 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.

[0087] 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.

[0088] 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.

[0089] In the present invention, when used, bornyl acetate or its salt, or a single agent or agricultural product prepared from a plant extract containing bornyl acetate or its salt is used to treat seeds, spray on leaves, or apply to roots.

[0090] In the present invention, bornyl acetate or its salt, or a plant extract containing bornyl acetate or its salt, or a product thereof is prepared into a solution for use in promoting seed germination or plant growth. The concentration of bornyl acetate or its salt, or a plant extract containing bornyl acetate or its salt in the solution can be selected according to actual needs.

[0091] 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.

[0092] The seeds are treated by mixing a single agent or an agricultural product made from bornyl acetate or its salts, or a plant extract containing bornyl acetate or its salts with the seeds, so that the bornyl acetate or its salts, or the plant extract containing bornyl acetate or its salts, adheres to or absorbs the seed surface.

[0093] Figures in the specification

[0094] Figure 1 shows the results of rice growth promotion; the left side shows the blank control, and the right side shows 20 ppm bornyl acetate;

[0095] Figure 2 shows the growth promotion results for corn; the left side shows the blank control, and the right side shows 20 ppm bornyl acetate;

[0096] Figure 3 shows the growth promotion results of Chinese cabbage; the left side shows the blank control, and the right side shows 20 ppm of bornyl acetate;

[0097] Figure 4 is a chromatogram of Cnidium monnieri essential oil;

[0098] FIG5 is a gas chromatography detection spectrum of valerian volatile oil;

[0099] FIG6 is a gas chromatography detection spectrum of valerian ethanol extract;

[0100] Figure 7 Bornyl acetate spectrum (standard);

[0101] Figure 8 volatile oil spectrum of Amomum villosum;

[0102] Figure 9 Spectrum of Houttuynia cordata volatile oil. DETAILED DESCRIPTION

[0103] 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.

[0104] Example 1 Bornyl acetate activity test

[0105] The wheat seedlings / corn seedlings / Chinese cabbage seedlings that were cultivated to a height of 10 cm were tested by leaf spraying. The leaves were sprayed according to the standard of moistening each leaf. After the treatment, they were placed in a culture room with a temperature of 22°C to 25°C and a humidity of 60% to 68%. The Chinese cabbage was treated with the drug twice throughout the entire process, with an interval of 7 days between each treatment. The corn and wheat were treated once throughout the entire process, with no less than 20 plants per treatment. The positive control was sprayed with 5000 times the recommended concentration of seaweed extract (10% powder, Haijingling, provided by the manufacturer in April 2022), the blank control was sprayed with an equal amount of water (CK), and borneol acetate was used at different concentrations (see the table below). After application, each treatment group was cultured under the same natural conditions.

[0106] When using seaweed extract, it is directly prepared with sterile water (the same below). When using bornyl acetate, it is prepared with sterile water, and an appropriate amount of ethanol (such as 1%) and surfactant (such as Tween 2%) are added to ensure that bornyl acetate is evenly dispersed in the water.

[0107] Survey Method: In the early stages of the experiment, all seedling conditions were kept consistent from the beginning of germination. Test materials with consistent growth and uniform size were selected. All treatment indicators were considered consistent in the early stages, and only the growth rate of each indicator after the drug application was calculated. A survey was conducted one week after the second application, and plant height, root length, leaf width, leaf thickness, stem diameter, SPAD, plant fresh weight, and root fresh weight were measured. The units for leaf width are mm, leaf thickness is mm, plant fresh weight is g, plant height is cm, root length is cm, stem diameter is mm, and root fresh weight is g.

[0108] Calculate the growth rate of each index after different treatments: Growth rate = (treatment index - blank index) * 100% / blank index.

[0109] The results of the index measurements of each treatment group 7-14 days after application are shown in the following table. The data in the table are all average values.

[0110] Experimental results:

[0111] (1) For Chinese cabbage

[0112] Table 1 Test results of different treatments for various indicators

[0113] Table 2 Test results of growth rate (%) of various indicators under different treatments

[0114] (2) For corn

[0115] Table 3 Test results of different treatments for various indicators

[0116] Table 4 Test results of growth rate (%) of various indicators under different treatments

[0117] (3) Cabbage

[0118] Table 5 Test results of different treatments for various indicators

[0119] Table 6 Test results of growth rate (%) of various indicators under different treatments

[0120] Example 2 Activity test of Cnidium monnieri volatile oil

[0121] The method comprises the following steps: taking Cnidium monnieri medicinal material, crushing the material, soaking and moistening the material with a steam distillation device, and reflux extraction until no more volatile oil is extracted; stopping heating and naturally cooling the material to room temperature; collecting the volatile oil from the volatile oil extraction device, and obtaining Cnidium monnieri volatile oil.

[0122] The components of different Cnidium monnieri essential oils were determined by gas chromatography using a DB-1 capillary column (100% dimethylpolysiloxane as the stationary phase) (30 m x 0.25 mm, 0.25 μm); column temperature 100°C, injection port temperature 230°C, detector (FID) temperature 250°C, and split ratio 10:1. The results are shown in Figure 4. Figure 4 shows the gas chromatography spectrum of Cnidium monnieri essential oils, showing a bornyl acetate content of 113,080.5001 μg / ml.

[0123] 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. When spraying the leaves, the medicine is sprayed according to the standard of moistening each leaf. After the treatment, the culture room is placed. 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. 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), the blank control is sprayed with an equal amount of water, and different concentrations of Cnidium monnieri volatile oil are used (see the table below). After application, each treatment group is cultured under the same natural conditions.

[0124] When using Cnidium monnieri essential oil, use sterile water to prepare it, and add appropriate amounts of ethanol (such as 5%) and surfactant (such as castor oil polyoxyethylene ether 5%) to ensure that the Cnidium monnieri essential oil is evenly dispersed in the water (follow this configuration when using other essential oils).

[0125] 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. Test materials with consistent growth 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.

[0126] The experimental treatment settings were as shown in Table 7 for foliar application. The results of the index measurements of each treatment group 7-14 days after application are shown in the following table. The data in the table are all average values.

[0127] Table 7 Experimental treatment settings

[0128] Table 8 The results of the index determination of each treatment group before and after the application of Chinese cabbage

[0129] Table 9 The results of the index determination of each treatment group before and after the application of Chinese cabbage

[0130] Table 10 Results of the indicators of each treatment group before and after the application of the corn

[0131] Table 11 Results of the indicators of each treatment group before and after the application of the corn

[0132] Table 6 Wheat index test results of each treatment group before and after drug application

[0133] Table 12 Wheat index test results of each treatment group before and after drug application

[0134] Example 3 Biological Activity of Valerian Volatile Oil

[0135] Volatile oil: Take valerian herbs, grind them, soak and moisten them in a steam distillation device, and reflux extract them until no more volatile oil is extracted. After stopping heating, naturally cool them to room temperature, and collect the volatile oil from the volatile oil extraction device to obtain valerian volatile oil.

[0136] Ethanol extract: Take an appropriate amount of valerian crude powder, use anhydrous ethanol at a solid-liquid ratio of 1:8, ultrasonically extract for 1 hour, extract twice, combine the two extracts, and concentrate to dryness under reduced pressure to obtain valerian ethanol extract.

[0137] The components of different valerian extracts were detected by gas chromatography, and 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; the split ratio was 10:1. The results are shown in Figures 1 and 2. Figure 5 is a gas chromatography detection spectrum of valerian volatile oil, and Figure 6 is a gas chromatography detection spectrum of valerian ethanol extract. Among them, the content of bornyl acetate in valerian volatile oil is 17150.3876μg / ml. As shown in Figure 6, valerian ethanol extract also contains bornyl acetate, and the content is lower than that of volatile oil.

[0138] 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, 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. Valerian volatile oil and ethanol extract are used at different concentrations (see the table below). After application, each treatment group is cultured under the same natural conditions.

[0139] 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. Test materials with consistent growth 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.

[0140] The experimental treatment settings were as shown in Table 13 for foliar application. The results of the index measurements of each treatment group 7-14 days after application are shown in the following table. The data in the table are all average values.

[0141] Table 13 Experimental treatment settings

[0142] Table 14 Results of index determination of each treatment group before and after application of Chinese cabbage

[0143] Table 15 The results of the index determination of each treatment group before and after the application of Chinese cabbage

[0144] Table 16 Results of the indicators of each treatment group before and after the application of corn

[0145] Table 17 Results of corn index determination in each treatment group before and after drug application

[0146] Table 18 Wheat index test results of each treatment group before and after drug application

[0147] Table 19 Wheat index test results before and after treatment

[0148] Example 4 Biological Activity of Amomum villosum Volatile Oil

[0149] Take Amomum villosum medicinal material, grind it, soak it and moisten it, use a steam distillation device to extract volatile oil until no more volatile oil is extracted, stop heating and naturally cool it to room temperature, collect the volatile oil from the volatile oil extraction device to obtain Amomum villosum volatile oil.

[0150] The components of the volatile oil were detected by gas chromatography using a DB-1 capillary column (100% dimethylpolysiloxane as the stationary phase) (30 m × 0.25 mm, 0.25 μm); column temperature 100°C, injection port temperature 230°C, detector (FID) temperature 250°C; split ratio 10:1.

[0151] FIG8 is a gas chromatography detection spectrum of Amomum villosum volatile oil, wherein the content of bornyl acetate is 334810.3980 μg / ml.

[0152] Test methods and investigation methods Reference Example 2

[0153] The experimental treatment settings are shown in Table 20 for foliar application. The results of the indicators of each treatment group 7-14 days after application are shown in the following table. The data in the table are all average values.

[0154] Table 20 Experimental treatment settings

[0155] Table 21 The results of the index determination of each treatment group before and after the application of Chinese cabbage

[0156] Table 22 Results of index determination of each treatment group before and after application of Chinese cabbage

[0157] Table 23 Results of indicators of each treatment group before and after corn application

[0158] Table 24 Results of the indicators of each treatment group before and after the application of corn

[0159] Example 5 Biological Activity of Houttuynia Cordata Volatile Oil

[0160] Take houttuynia cordata herb, crush it, soak it and moisten it, use a steam distillation device to extract volatile oil until no more volatile oil is extracted, stop heating and naturally cool it to room temperature, collect the volatile oil from the volatile oil extraction device to obtain houttuynia cordata volatile oil.

[0161] The components of the volatile oil were detected by gas chromatography using a DB-1 capillary column (100% dimethylpolysiloxane as the stationary phase) (30 m × 0.25 mm, 0.25 μm); column temperature 100°C, injection port temperature 230°C, detector (FID) temperature 250°C; split ratio 10:1.

[0162] FIG9 is a gas chromatography detection spectrum of Houttuynia cordata volatile oil, wherein the content of bornyl acetate is 18600.8279 μg / ml.

[0163] Test methods and investigation methods Reference Example 2

[0164] The experimental treatment settings were as shown in Table 25 for foliar application. The results of the indicators of each treatment group were measured 7-14 days after application and are shown in the following table. The data in the table are all average values.

[0165] Table 25 Experimental treatment settings

[0166] Table 26 Results of index determination of each treatment group before and after application of Chinese cabbage

[0167] Table 27 The results of the index determination of each treatment group before and after the application of Chinese cabbage

[0168] Table 28 Results of indicators of each treatment group before and after corn application

[0169] Table 29 Results of index determination of each treatment group before and after corn application

Claims

1. Use of borneol acetate or its salts or isomers, or plant extracts containing borneol acetate or its salts or isomers in the preparation of biostimulants.

2. The use according to claim 1, characterized in that: Application of the borneol acetate or its salts and isomers, or plant extracts containing borneol acetate or its salts and isomers in promoting plant growth.

3. The use according to claim 1, characterized in that: The plant extract comprises one or a mixture of multiple kinds of Cnidium monnieri extract, Amomum villosum extract, Houttuynia cordata extract, Siberian pine needle oil, Artemisia argyi volatile oil, Forsythia suspensa volatile oil and Cinnamomum cassia twig volatile oil.

4. The use according to claim 3, characterized in that: The Cnidium monnieri extract is selected from Cnidium monnieri volatile oil or an extract containing Cnidium monnieri volatile oil; the Amomum villosum extract is selected from Amomum villosum volatile oil or an extract containing Amomum villosum volatile oil; the Houttuynia cordata extract is selected from Houttuynia cordata volatile oil or an extract containing Houttuynia cordata volatile oil.

5. The use according to claim 3, characterized in that: The plant extract contains borneol acetate at more than 1 mg / ml.

6. The use according to claim 3, characterized in that: The plant extract contains 1-1000 mg / ml of bornyl acetate.

7. The use according to any one of claims 1 to 6, 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.

8. The use according to claim 2, characterized in that: Plants whose growth is promoted include cruciferous vegetables, field crops, fruit trees, and nightshades.

9. The use according to claim 1, characterized in that: When in use, borneol acetate or its salt, or a plant extract containing borneol acetate or its salt is made into an agricultural product for use, 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.

10. The use according to claim 9, characterized in that: The dosage forms of the product are emulsifiable concentrate, suspension, wettable powder, powder, granule, aqueous solution, mother liquor, mother powder, microemulsion, microcapsule suspension, aqueous emulsion, tablet, smoke agent, sustained-release agent and seed coating agent.

11. The use according to claim 9, characterized in that: The agricultural product made of borneol acetate or its salt, or a plant extract containing borneol acetate or its salt is used in combination with foliar fertilizer, water-soluble fertilizer, compound fertilizer, pesticide, or biostimulant.

12. The use according to claim 11, characterized in that: When used, the borneol acetate or its salt, or a single agent or a prepared agricultural product containing the borneol acetate or its salt is used to treat soil, seeds, spray on leaves or irrigate roots.

13. A biostimulant or plant growth promoter, characterized in that: The active ingredient of the invention is (1) borneol acetate or its salt or isomer; or (2) a plant extract containing borneol acetate or its salt or isomer.

14. The biostimulant or plant growth promoter according to claim 13, characterized in that: The plant extract is selected from one or a mixture of multiple kinds of Cnidium monnieri extract, Amomum villosum extract, Houttuynia cordata extract, Siberian pine needle oil, Artemisia argyi volatile oil, Forsythia suspensa volatile oil, and Cinnamomum cassia twig volatile oil.

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