A composition containing phenolic compounds and its application

Abstract

The present invention relates to a novel plant regulator composition, specifically a composition containing a phenolic compound and its application. The composition comprises a phenolic compound and a cytokinin-like substance, the mass ratio of the two being 1:30-30:1; wherein the phenolic compound is 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol, the molecular formula of which is C 18 H 24 O, relative molecular mass: 256.4, structural formula is as follows, #imgabs0#The present invention significantly improves the efficiency of light energy utilization: Psoralea corylifolia seed extract can enhance the light-capturing ability of plant chloroplasts, synergize with cytokinin substances, significantly improve the efficiency of light energy utilization of plants, promote plant photosynthesis, thereby synthesizing more organic matter, and providing sufficient energy and material basis for plant growth and development.

Description

Technical Field

[0001] The present invention relates to a novel plant regulator composition, in particular to a composition containing phenolic compounds and its application. Background Art

[0002] Cytokinins are plant hormones whose primary function is to promote cell division and differentiation. Due to nutrient deficiencies, their use alone cannot meet the nutritional requirements of plant growth, potentially negatively impacting crop yields.

[0003] Psoralea corylifolia seed extract is a botanical biopesticide product with pesticide registration number PD20190058, pesticide production license number Pesticide Shengxu (Liao) 0030, and product standard number Q / SYTX 009-2023. Although Psoralea corylifolia seed extract and its component compounds have potential for controlling plant diseases and promoting plant growth, the chemical composition of Psoralea corylifolia seeds is extremely complex, with unknown active substances. Different extraction methods can lead to differences in active substances, and the activity and target sites of various compounds vary. Furthermore, the effects of combining this extract with other substances cannot be accurately predicted. Summary of the Invention

[0004] The purpose of the present invention is to provide a composition containing phenolic compounds and application thereof.

[0005] To achieve the above purpose, the present invention adopts the following technical solutions:

[0006] A composition containing a phenolic compound, comprising a phenolic compound and a cytokinin-like substance, wherein the mass ratio of the phenolic compound and the cytokinin-like substance is 1:30-30:1; wherein the phenolic compound is 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol, and the molecular formula is C 18 H 24 O, relative molecular mass: 256.4, structural formula is as follows,

[0007]

[0008] The above-mentioned phenolic compounds are obtained by weighing psoralea corylifolia seeds or seed powder, adding water with a mass volume ratio of g:mL of 1:10-20, 20%, 40%, 60% or 80% or 90% ethanol (preferably 60% ethanol with a mass volume ratio of g:mL of 1:10-20), extracting at 25-100 ° C for 1-4 hours, pouring out the extract, repeating the extraction 2-4 times, combining the extracts, and concentrating the extract under reduced pressure to a density of 1.0 g / cm 3The crude extract was mixed with distilled water in a 1:1 volume ratio to obtain a suspension. An equal volume of ethyl acetate was then added to the suspension and mixed thoroughly. The mixture was allowed to stand and separate into two layers. The ethyl acetate extract was extracted. This process was repeated five times. The ethyl acetate extracts were combined and concentrated under vacuum at 45°C until the extract was in the form of an extract, thereby obtaining the purified Psoralea corylifolia seed extract. The extract was analyzed by analytical high-performance liquid chromatography (HPLC). The chromatographic column was a Hypersil ODS column (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of a 30:70 volume ratio of methanol and 0.1% glacial acetic acid aqueous solution. The flow rate was 1 mL / min, the column temperature was 28°C, and the injection volume was 10 μL. A full scan of the analytical HPLC analysis of the sample confirmed that the maximum absorption wavelength of the purified Psoralea corylifolia seed extract was 262 nm. The purified Psoralea corylifolia seed extract was dissolved in methanol at a ratio of 1:10 (g:mL) and further separated and purified by preparative liquid chromatography. The preparation conditions were: chromatographic column (C18-8), mobile phase of methanol-water (30:70), injection volume of 10mL, flow rate of 20mL / min, detection wavelength of 262nm, and retention time of 12.422min. Pure 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol was obtained, with a content of 97.51%. The substance appears as a light yellow viscous liquid with an empirical formula of: C 18 H 24 O, relative molecular mass: 256.4, structural formula:

[0009]

[0010] The substance was characterized by nuclear magnetic resonance spectroscopy and mass spectrometry. Figure 1 and Figure 2 The liquid chromatogram of content determination is shown in Figure 3 .

[0011] The cytokinin substances are one or more of furfurylaminopurine, benzylaminopurine and hydroxyeneadenine.

[0012] An application of the composition containing phenolic compounds, and an application of the composition in the preparation of a plant growth regulator.

[0013] The dosage form of the regulator is a soluble agent, a wettable powder, a suspension, a soluble powder, a water-dispersible granule, an emulsifiable concentrate, a microcapsule suspension or a microemulsion.

[0014] The preferred dosage form is a soluble solution. Specifically:

[0015] The composition is dissolved in a solvent such as ethanol, glycerol, dimethyl sulfoxide or water, and one or more of T-20, NP-15, NP-7, EL-40, ZSL-6140, S-80, Nongru 500, Nongru 601, Nongru 1600 or T-80 is added to prepare a pesticide soluble solution.

[0016] The composition is used in regulating the growth of fruit trees, vegetables or grain crops, increasing production and income, and improving fruit quality.

[0017] Advantages and effects of the present invention:

[0018] (1) Significantly improve light energy utilization efficiency: Psoralea corylifolia seed extract can enhance the light capture ability of plant chloroplasts, synergize with cytokinin-like substances, significantly improve the light energy utilization efficiency of plants, promote plant photosynthesis, and thus synthesize more organic matter, providing sufficient energy and material basis for plant growth and development.

[0019] (2) Promote plant growth and development: Psoralea corylifolia seed extract is rich in a variety of nutrients required for plant growth. When mixed with cytokinin-like substances, it can accelerate cell division, promote plant growth and development, make the plant root system well-developed, the stems thick, and the leaves dark green, and improve the plant's resistance to stress and disease and pests.

[0020] (3) Achieve increased production and harvest: By improving the efficiency of light energy utilization and promoting plant growth and development, the mixed use of Psoralea corylifolia seed extract and cytokinin substances can significantly improve the yield and quality of plants and achieve the goal of increased production and harvest.

[0021] (4) Reduce fertilizer use and lower costs: Psoralea corylifolia seed extract is rich in various nutrients required for plant growth. When mixed with cytokinin-like substances, it can reduce the use of chemical fertilizers, reduce agricultural production costs, and at the same time reduce the pollution of chemical fertilizers to the environment, thereby achieving sustainable agricultural development. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 1H NMR spectrum of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol provided in an embodiment of the present invention.

[0023] Figure 2 The mass spectrum of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol provided in an embodiment of the present invention.

[0024] Figure 3 The liquid chromatogram of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0025] The specific embodiments of the present invention are further described below with reference to examples. It should be noted that the specific embodiments described herein are only for the purpose of illustrating and explaining the present invention, and are not intended to limit the present invention.

[0026] Example 1 Preparation of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol

[0027] Weigh 100g of Psoralea corylifolia seeds, add 1000mL of 60% ethanol, extract at 100°C for 4 hours, pour out the extract, repeat the extraction 3 times, combine the extracts, and concentrate the extracts under reduced pressure to a crude extract with a density of 1.0g / cm3. The crude extract is mixed and suspended with distilled water in a volume ratio of 1:1, and then an equal volume of ethyl acetate is added to the suspension and mixed thoroughly. The mixture is allowed to stand and separate into two layers. The ethyl acetate extract phase is taken, and the operation is repeated 5 times. The ethyl acetate extract phases are combined and concentrated under vacuum at 45°C to an extract form to obtain a purified Psoralea corylifolia seed extract.

[0028] Analytical high performance liquid chromatography (HPLC) was used for analysis. The chromatographic column was a Hypersil ODS column (250 mm × 4.6 mm, 5 μm), the mobile phase was a 30:70 volume ratio of methanol and 0.1% glacial acetic acid aqueous solution, the flow rate was 1 mL / min, the column temperature was 28°C, and the injection volume was 10 μL. A full scan of the analytical HPLC was performed on the substance in Example 1, and the maximum absorption wavelength of the substance was determined to be 262 nm. The extract obtained above was dissolved in methanol at a ratio of 1:10 (g / ml) and further separated and purified by preparative HPLC. The preparative conditions were: chromatographic column (C18-8), mobile phase of methanol-water (30:70), injection volume of 10 mL, flow rate of 20 mL / min, detection wavelength of 262 nm, and retention time of 12.422 min. The pure product of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol is obtained, with a content of 97.51%. Appearance: light yellow viscous liquid, molecular formula: C18H24O, relative molecular mass: 256.4, structural formula:

[0029]

[0030] The substance was characterized by nuclear magnetic resonance spectroscopy and mass spectrometry. Figure 1 and Figure 2 The liquid chromatogram of content determination is shown in Figure 3 .

[0031] Example 2 Preparation of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol-furfurylaminopurine soluble solution

[0032] Weigh 1.0 g of the phenolic substance obtained in Example 1, add 5 g of furfurylaminopurine, 2 g of ethanol, and 2 g of dimethyl sulfoxide, stir until completely dissolved, then add 3 g of T-80, 0.5 g of Nongru 1600, and 12 g of EL-40, continue stirring until completely mixed, and finally add water to make up to 100 g, stir evenly to obtain 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol.furfurylaminopurine soluble liquid preparation.

[0033] Example 3 Preparation of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol-benzylaminopurine soluble solution

[0034] Weigh 1 g of the phenolic substance obtained in Example 1, add 3 g of benzylaminopurine, 2 g of ethanol, and 1 g of dimethyl sulfoxide, stir until completely dissolved, then add 3 g of T-20 and 12 g of ZSL-6140, continue stirring until completely mixed, and finally add water to make up to 100 g, stir evenly to obtain 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol.benzylaminopurine soluble solution.

[0035] Example 4 Preparation of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol-hydroxyene adenine soluble preparation

[0036] 1.2 g of the phenolic substance obtained in Example 1 was weighed, 3 g of hydroxyene adenine, 2 g of propylene glycol, and 1 g of dimethyl sulfoxide were added, and the mixture was stirred until completely dissolved. Then, 3 g of T-80, 2 g of NP-7, and 12 g of EL-40 were added, and stirring was continued until completely mixed. Finally, water was added to make up to 100 g, and the mixture was stirred evenly to obtain a 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol / hydroxyene adenine soluble solution.

[0037] Example 5 Preparation of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol soluble solution

[0038] Weigh 1 g of the phenolic substance obtained in Example 1, add 2 g of ethanol, stir until completely dissolved, then add 3 g of T-20 and 12 g of ZSL-6140, continue stirring until completely mixed, and finally add water to make up to 100 g, and stir evenly to obtain a soluble solution of 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol.

[0039] Application Example 1: Experiment on the impact on tobacco growth and yield

[0040] 1. Trial Location

[0041] The experiment was carried out in the Xianyang experimental field in Pucheng, Nanping City, Fujian Province. The soil is loose and fertile, with a pH value of 7.3 and medium soil fertility.

[0042] 2. Test Materials

[0043] The test crop was tobacco, variety: Yunyan 87. The test agents were the substances of Example 2 and Example 5.

[0044] 3. Experimental Design

[0045] The experiment included four treatments: Treatment 1: Example 2 formulation at a concentration of 300 mg / L; Treatment 2: Example 5 formulation at a concentration of 300 mg / L; Treatment 3: Spraying of furfural purine powder (commercially available) at a concentration of 1000 mg / L; and Treatment 4 (CK): Spraying of water. Applications were made three times, starting at the two-leaf, one-heart stage of the tobacco plant, with one application every 30 days.

[0046] 4. Survey items and methods

[0047] When harvesting tobacco leaves from different parts, 10 leaves from the corresponding harvested parts were randomly selected from each plot to measure the leaf length and leaf width, and the average value was calculated. After the tobacco leaves from different parts were harvested and cured, 30 leaves from the corresponding harvested parts were randomly selected from each plot to weigh the total weight of the flue-cured tobacco, and the average value was calculated to measure the weight of a single flue-cured tobacco leaf. Fresh tobacco leaves from all parts of the tobacco leaves were harvested in the whole plot and conventionally cured into flue-cured tobacco. The flue-cured tobacco yield was calculated, and the yield was expressed as kg / 667m 2 express.

[0048] The calculation formula is as follows:

[0049] Leaf length (width) regulation rate = [leaf length (width) of the drug-treated area - leaf length (width) of the water control area] / leaf length (width) of the water control area x 100%;

[0050] Single leaf mass increase rate = (single leaf mass in the drug-treated area - single leaf mass in the water control area) / single leaf mass in the water control area x 100%;

[0051] Yield increase rate = (yield per mu in the chemical treatment area - yield per mu in the water control area) / yield per mu in the water control area x 100%

[0052] When the upper tobacco leaves were harvested, 5 fresh tobacco leaf samples were randomly selected from each plot and placed in an electric forced air drying oven. They were baked at a constant temperature of 70°C for 48 h to make flue-cured tobacco samples. 10 fresh upper tobacco leaf samples were randomly selected from each plot, bagged and marked. The water-soluble total sugar content and protein content of the fresh tobacco leaves were determined according to NY / T3030-2016 "Determination of the Content of Total Water-Soluble Sugar in Cotton - Anthrone Colorimetric Method" and GB 5009.5-2016 "National Food Safety Standard - Determination of Protein in Food", and the nicotine content in the flue-cured tobacco samples was determined by spectrophotometry.

[0053] 5. Results and Analysis

[0054] Table 1 Effects of different treatments on tobacco leaf length (width)

[0055]

[0056] As can be seen from the above table, each treatment has a certain promoting effect on tobacco leaf length and leaf width, among which treatment 1 has the best promoting effect.

[0057] Table 2 Effects of different treatments on tobacco leaf quality and yield

[0058]

[0059] As can be seen from the above table, each treatment has a certain promoting effect on the quality and yield of tobacco leaves, among which treatment 1 has the best promoting effect.

[0060] Table 3 Effects of different treatments on tobacco quality

[0061]

[0062]

[0063] It can be seen from the above table that different treatments have no adverse effects on tobacco quality.

[0064] Application Example 2 Experiment on the Effect of Wheat Growth and Yield

[0065] 1. Trial Location

[0066] The experimental site is located in Airport Village, Liufenchang, Dashahu Management District, Honghu City, Hubei Province. The area of the experimental site is 28 mu. The soil type is light loam, pH value is 7.0, organic matter content is 1.8%, and the previous crop is soybean.

[0067] 2. Test Materials

[0068] The test crop was wheat, variety: Huamai 1598. The test agents were the substances of Example 3 and Example 5.

[0069] 3. Experimental Design

[0070] The experiment included four treatments: Treatment 1: Example 3 formulation at a concentration of 100 mg / L; Treatment 2: Example 5 formulation at a concentration of 200 mg / L; Treatment 3: Benzylaminopurine powder (commercially available) at a concentration of 1000 mg / L; and Treatment 4 (CK): spraying with water. Applications were made three times each at the wheat's greening stage, heading stage, and grain filling stage.

[0071] 4. Survey items and methods

[0072] During the wheat harvest period, the wheat plant height, effective ears per unit area, and plot yield are investigated, and samples are taken for quality analysis. At the same time, indoor seed sampling is carried out, and after drying, the number of filled grains in the wheat ears and the thousand-grain weight are measured.

[0073] Plant height survey: Samples were taken at 5 points in each plot, with 4 plants at each point, to measure the wheat plant height.

[0074] Effective ear survey: sampling at 5 points in each plot, 0.25m per point 2 , investigate the effective ears of wheat.

[0075] Yield survey: Each plot is harvested individually and the yield is measured. The yield statistics are in kg.667m -2 express.

[0076] Indoor seed testing sampling: sampling was conducted at five points in each plot, with 20 plants at each point. The samples were brought back to the laboratory to investigate the number of grains per ear and the thousand-grain weight.

[0077] Wheat quality determination: About 500 g of wheat samples were collected from each treatment plot and sent to a third party for testing of starch, soluble sugar, and protein content.

[0078] 5. Results and Analysis

[0079] Table 4 Effects of different treatments on wheat growth and yield

[0080]

[0081] As can be seen from the above table, Treatment 1 and Treatment 2 can effectively promote wheat growth, increase the number of effective ears, number of grains per ear and 1000-grain weight, etc., and the yield-increasing effect is obvious.

[0082] Table 5 Effects of different treatments on wheat quality

[0083]

[0084] As can be seen from the above table, treatment 1 and treatment 2 can effectively improve wheat quality.

[0085] Application Example 3: Experiment on the impact on soybean growth and yield

[0086] 1. Trial Location

[0087] The experimental site is located in Jixiang Village, Lindian County, Daqing City, Heilongjiang Province. The area of the experimental site is 20 mu. The soil type is light loam, pH value is 7.9, organic matter content is 3.1%, and the previous crop is cabbage.

[0088] 2. Test Materials

[0089] The test crop was soybean, variety: Heihe No. 43. The test agents were the substances of Example 4 and Example 5.

[0090] 3. Experimental Design

[0091] The experiment included five treatments: Treatment 1: Preparation of Example 4 at a concentration of 200 mg / L; Treatment 2: Preparation of Example 5 at a concentration of 300 mg / L; Treatment 3: Hydroxyphenadenine powder (commercially available) sprayed at a concentration of 1000 mg / L; Treatment 4: Psoralea corylifolia plant nutrient solution prepared by diluting the extract of Psoralea corylifolia seeds obtained in Example 1 3000-fold with water; Treatment 5 (CK): Sprayed with plain water. Applications were made once each at the branching and pod-setting stages, for a total of two applications. Other field management procedures were the same as those for field production.

[0092] Table 6 Effects of different treatments on soybean growth and yield

[0093]

[0094] As can be seen from the above table, treatment 3 and other treatments can significantly promote soybean growth and increase soybean yield.

Claims

1. A composition containing a phenolic compound, characterized in that: The composition comprises a phenol compound and a cytokinin substance, the mass ratio of the two being 1:30-30:1; wherein the phenol compound is 4-(2-(methyl-2-(4-methylpent-3-en-1-yl)cyclopent-3-en-1-yl)phenol, the molecular formula of which is C 18 H 24 O, relative molecular mass: 256.4, structural formula is as follows, ； The cytokinin substances are one or more of furfurylaminopurine, benzylaminopurine and hydroxyeneadenine.

2. An application of the composition containing phenolic compounds according to claim 1, characterized in that: Application of the composition in regulating plant growth, increasing production and income, and improving fruit quality; The plants are tobacco, wheat and soybean.

3. The use of the composition containing phenolic compounds according to claim 2, characterized in that: The composition dosage form is a soluble agent, a wettable powder, a suspension, a soluble powder, a water-dispersible granule, an emulsifiable concentrate, a microcapsule suspension or a microemulsion.

Images