Chemical control agent suitable for liquorice and screening method of chemical control agent

Abstract

The invention relates to a chemical control agent suitable for liquorice and screening of the chemical control agent. The chemical control agent is composed of uniconazole. The invention also provides application of uniconazole in preparation of the glycyrrhiza chemical control agent, and optionally, the chemical control agent has one or more of the following applications: a, reducing the area of glycyrrhiza leaves; b, increasing the chlorophyll content of licorice leaves; c, thickening the roots of the licorice roots; d, reducing the height of licorice plants; e, increasing underground biomass of liquorice; f, increasing the content of total flavonoids in liquorice roots; and g, increasing the content of one or more of glycyrrhizic acid, liquiritigenin, isoliquiritigenin and glycyrrhetinic acid of the licorice roots. The invention further provides a method for improving lodging resistance of liquorice. The method comprises the step of applying uniconazole to the liquorice. The invention provides a thought and a method for effectively controlling the plant height of liquorice and improving the economic benefit of liquorice cultivation, and lays a foundation for field management and sustainable development of subsequent liquorice cultivation.

Description

Technical Field

[0001] The invention belongs to the field of agricultural biotechnology, and particularly relates to a chemical control agent suitable for licorice and its screening. Background Art

[0002] Crop chemical control refers to an agricultural technique that uses chemical substances, such as plant growth regulators, to alter the plant's endogenous hormone system and crop growth and development processes to increase crop yield, improve quality, and enhance stress resistance. Chemical regulation is an important supplement to traditional agricultural production methods and a key measure for high-yield cultivation. By regulating plant growth and development, it is widely used in agriculture and horticulture to increase yield, improve quality, and enhance stress resistance. Currently, the most commonly used chemical regulators include uniconazole, paclobutrazol, chlormequat chloride, prohexadione-calcium, and mepiquat. Using appropriate chemical control agents can solve many problems in field crop production and is a key technology for improving land productivity and achieving agricultural modernization.

[0003] With the rapid development of chemical crop control technologies, chemical control agents have found widespread application in agriculture, horticulture, and other fields. They play a vital role in everything from grain crops like wheat, corn, cotton, rice, and soybeans, to vegetables and fruits like tomatoes, cucumbers, and grapes, to fruit preservation and the cultivation of tea and flowers. Research has shown that spraying appropriate concentrations of chemical control agents like paclobutrazol during the soybean V4 stage can influence plant hormone balance, inhibit the synthesis or action of gibberellins, thereby slowing cell elongation and shortening internodes, resulting in dwarfing and ultimately regulating soybean plant shape. Spraying uniconazole during the early jointing stage of rice not only improves the morphology and mechanical properties of high-quality rice stems, enhancing their resistance to lodging, but also significantly increases the yield of taller varieties. The widespread application of chemical control technologies on crops has played a significant role in increasing crop yields, improving mechanized harvesting, and improving crop quality, creating significant economic and social benefits for modern agriculture.

[0004] Uniconazole is a new, highly effective plant growth regulator with a bioactivity 6-10 times higher than that of paclobutrazol. Its unique mechanism of action, as a gibberellin synthesis inhibitor, inhibits pathogen growth by hindering ergosterol biosynthesis. Therefore, it possesses both growth control and antibacterial properties. During plant growth, Uniconazole primarily inhibits internode elongation by regulating the volume, length, and intercellular arrangement of individual cells, thereby dwarfing and compacting the plant. It also effectively promotes lateral bud growth and flower bud formation, and has effects on photosynthesis and antioxidant systems, laying the foundation for healthy plant development. Uniconazole effectively promotes tillering in wheat, enhances lodging resistance, significantly increases ear formation, results in dark green leaves, high photosynthetic efficiency, a well-developed root system, and improved seedling quality. Spraying Uniconazole on rice seedlings during the seedling stage effectively regulates growth, reducing seedling height and increasing tillering, resulting in strong, multi-tilling seedlings and significantly increasing yield. Uniconazole plays an important role in the growth of plants. In the agricultural production process, appropriate concentrations of Uniconazole or other chemical control agents can be used to meet the needs of agricultural production, providing a solid theoretical basis for its wide application in agricultural production.

[0005] Licorice is a major traditional Chinese medicinal herb and a pioneering plant for windbreaks and sand fixation in the desert regions of northwest China. Currently, overexploitation of wild licorice resources has led to a shortage of reserves, leaving cultivated licorice as the primary market supply. However, a current problem with cultivated licorice is that the plants grow too tall, reaching heights exceeding 150 cm. This results in poor lodging resistance in adverse weather conditions, impacting field management and root development. Research has shown that chemical control agents can control plant height and improve plant yield and quality. However, no chemical control agents suitable for licorice are currently available. Summary of the Invention

[0006] The present invention selects five chemical control agents commonly used in agricultural production and uses an exogenous spraying method to treat licorice to screen out chemical control agents suitable for artificial cultivation of licorice. Liquid chromatography-mass spectrometry (UPLC-MS) is used to analyze the effect of uniconazole on the content of active ingredients in Ural Glycyrrhiza uralensis. This provides ideas and methods for effectively controlling licorice plant height and improving the economic benefits of cultivated licorice, and lays a foundation for subsequent field management and sustainable development of cultivated licorice.

[0007] Specifically, the present invention provides the following technical solutions.

[0008] In one aspect, the present invention provides a glycyrrhizic acid control agent consisting of uniconazole.

[0009] On the other hand, the present invention provides a chemical control agent for improving the lodging resistance of licorice, which consists of uniconazole.

[0010] In another aspect, the present invention provides use of uniconazole in preparing a glycyrrhizic acid control agent.

[0011] In some embodiments, the chemical control agent has one or more of the following uses:

[0012] a. Reduce the leaf area of licorice;

[0013] b. Increase the chlorophyll content of licorice leaves;

[0014] c. Make the roots of licorice thicker;

[0015] d. Reduce the height of licorice plants;

[0016] e. Increase the underground biomass of licorice;

[0017] f. Increase the total flavonoid content in licorice root;

[0018] g. Increase the content of one or more of glycyrrhizic acid, glycyrrhizin, isoliquiritigenin and glycyrrhetinic acid in licorice root.

[0019] In another aspect, the present invention provides a method for chemically controlling licorice, comprising applying uniconazole to the licorice, wherein the chemically controlling method can achieve one or more of the following uses:

[0020] a. Reduce the leaf area of licorice;

[0021] b. Increase the chlorophyll content of licorice leaves;

[0022] c. Make the roots of licorice thicker;

[0023] d. Reduce the height of licorice plants;

[0024] e. Increase the underground biomass of licorice;

[0025] f. Increase the total flavonoid content in licorice root;

[0026] g. Increase the content of one or more of glycyrrhizic acid, glycyrrhizin, isoliquiritigenin and glycyrrhetinic acid in licorice root.

[0027] In another aspect, the present invention provides a method for improving the lodging resistance of licorice, comprising applying uniconazole to the licorice.

[0028] In another aspect, the present invention provides a method for increasing the underground biomass of licorice, comprising applying uniconazole to licorice.

[0029] In another aspect, the present invention provides a method for increasing the accumulation of active ingredients in licorice, comprising administering uniconazole to licorice.

[0030] In some embodiments, the pharmaceutically active ingredient includes one or more of glycyrrhizic acid, glycyrrhizin, isoliquiritigenin, and glycyrrhetinic acid.

[0031] In some embodiments, the amount of uniconazole used is 80 mg / L to 250 mg / L.

[0032] In some embodiments, the amount of uniconazole is 80 mg / L to 240 mg / L, 80 mg / L to 230 mg / L, 80 mg / L to 220 mg / L, 80 mg / L to 210 mg / L, 80 mg / L to 200 mg / L, 80 mg / L to 190 mg / L, 80 mg / L to 180 mg / L, 80 mg / L to 170 mg / L, 80 mg / L to 160 mg / L, 80 mg / L to 150 mg / L, 80 mg / L to 140 mg / L, 80 mg / L to 130 mg / L, 80 mg / L to 120 mg / L, 80 mg / L to 110 mg / L, 80 mg / L to 100 mg / L, 80 mg / L to 90 mg / L, 90 mg / L to 250 mg / L, 90 mg / L to 240mg / L, 90mg / L to 230mg / L, 90mg / L to 220mg / L, 90mg / L to 210mg / L, 90mg / L to 200mg / L, 90mg / L to 190mg / L, 90mg / L to 180mg / L, 90mg / L to 170mg / L, 90mg / L to 160mg / L, 90mg / L to 150mg / L, 90mg / L to 140mg / L, 90mg / L to 130mg / L, 90mg / L to 120mg / L, 90mg / L to 110mg / L, 90mg / L to 100mg / L, 100 100mg / L to 150mg / L, 100mg / L to 140mg / L, 100mg / L to 130mg / L, 100mg / L to 120mg / L, 100mg / L to 210mg / L, 100mg / L to 200mg / L, 100mg / L to 190mg / L, 100mg / L to 180mg / L, 100mg / L to 170mg / L, 100mg / L to 160mg / L, 100mg / L to 150mg / L, 100mg / L to 140mg / L, 100mg / L to 130mg / L, 100mg / L to 120mg / L, 100mg / L to 110mg / L, 110 mg / L to 250mg / L, 110mg / L to 240mg / L, 110mg / L to 230mg / L, 110mg / L to 220mg / L, 110mg / L to 210mg / L, 110mg / L to 200mg / L, 110mg / L to 190mg / L, 110mg / L to 180mg / L, 110mg / L to 170mg / L, 110mg / L to 160mg / L, 110mg / L to 150mg / L, 110mg / L to 140mg / L, 110mg / L to 130mg / L, 110mg / L to 120mg / L, 120mg / L to 250mg / L, 120mg / L to 240mg / L, 120mg / L to 230mg / L, 120mg / L to 220mg / L, 120mg / L to 210mg / L, 120mg / L to 200mg / L, 120mg / L to 190mg / L, 120mg / L to 180mg / L, 120mg / L to 170mg / L, 120mg / L to 160mg / L, 120mg / L to 150mg / L, 120mg / L to 140mg / L, 120mg / L to 130mg / L, 130 mg / L to 250mg / L, 130mg / L to 240mg / L, 130mg / L to 230mg / L, 130mg / L to 220mg / L, 130mg / L to 210mg / L, 130mg / L to 200mg / L, 130mg / L to 190mg / L, 130mg / L to 180mg / L, 130mg / L to 170mg / L, 130mg / L to 160mg / L, 130mg / L to 150mg / L, 130mg / L to 140mg / L, 140 mg / L to 250mg / L, 140mg / L to 240mg / L, 140mg / L to 230mg / L, 140mg / L to 220mg / L, 140mg / L to 210mg / L, 140mg / L to 200mg / L, 140mg / L to 190mg / L, 140mg / L to 180mg / L, 140mg / L to 170mg / L, 140mg / L to 160mg / L, 140mg / L to 150mg / L, 150 mg / L to 250mg / L, 150mg / L to 240mg / L, 150mg / L to 230mg / L, 150mg / L to 220mg / L, 150mg / L to 210mg / L, 150mg / L to 200mg / L, 150mg / L to 190mg / L, 150mg / L to 180mg / L, 150mg / L to 170mg / L, 150mg / L to 160 mg / L, 160mg / L to 250mg / L, 160mg / L to 240mg / L, 160mg / L to 230mg / L, 160mg / L to 220mg / L, 160mg / L to 210mg / L, 160mg / L to 200mg / L, 160mg / L to 190mg / L, 160mg / L to 180mg / L, 160mg / L to 170mg / L, 170mg / L to 250mg / L, 170mg / L to 240mg / L, 170mg / L to 230mg / L, 170mg / L to 220mg / L, 170mg / L to 210mg / L, 170mg / L to 200mg / L, 170mg / L to 190mg / L, 170mg / L to 180mg / L, 180 mg / L to 250mg / L, 180mg / L to 240mg / L, 180mg / L to 230mg / L, 180mg / L to 220mg / L, 180mg / L to 210mg / L, 180mg / L to 200mg / L, 180mg / L to 190mg / L, 190 mg / L to 250mg / L, 190mg / L to 240mg / L, 190mg / L to 230mg / L, 190mg / L to 220mg / L, 190mg / L to 210mg / L, 190mg / L to 200mg / L, 200mg / L to 250mg / L, 200mg / L to 240mg / L, 200mg / L to 230mg / L, 200mg / L to 220mg / L, 200mg / L to 210mg / L, 210mg / L to 250mg / L, 210mg / L to 240mg / L, 210mg / L to 230mg / L, 210mg / L to 220mg / L, 220 mg / L to 250 mg / L, 220 mg / L to 240 mg / L, 220 mg / L to 230 mg / L, 230 mg / L to 250 mg / L, 230 mg / L to 240 mg / L, or 240 mg / L to 250 mg / L.

[0033] In some embodiments, the amount of uniconazole is 80 mg / L, 90 mg / L, 100 mg / L, 110 mg / L, 120 mg / L, 130 mg / L, 140 mg / L, 150 mg / L, 160 mg / L, 170 mg / L, 180 mg / L, 190 mg / L, 200 mg / L, 210 mg / L, 220 mg / L, 230 mg / L, 240 mg / L or 250 mg / L.

[0034] In some embodiments, the amount of uniconazole used is 250 mg / L.

[0035] In some embodiments, the licorice is Glycyrrhiza uralensis.

[0036] In some embodiments, the uniconazole is applied by spraying on licorice leaves.

[0037] In some embodiments, the uniconazole is applied by spraying on licorice leaves in the morning before sunrise.

[0038] In some embodiments, the uniconazole is applied by spraying on the leaves of licorice in the morning before the sun rises during the licorice seedling stage.

[0039] definition

[0040] Paclobutrazol: a chemical control agent belonging to the triazole class of compounds. Its chemical name is (2RS,3RS)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)pentan-3-ol, and its molecular formula is C 15 H 20 ClN3O. It can inhibit the biosynthesis of gibberellins in plants, thereby slowing down plant growth, and has the effects of controlling shoots, promoting flowering, dwarfing plants, and improving stress resistance.

[0041] Chlormequat: also known as choline chloride, choline chloride, chlormequat, etc., the chemical name is 2-chloroethyltrimethylammonium chloride, the molecular formula is C5H 13 Cl2N is a gibberellin antagonist that can enter the plant through leaves, young branches, buds, roots, and seeds. Its mechanism of action is to inhibit the biosynthesis of gibberellins in the plant, mainly by inhibiting the production of kaurene, thereby hindering the biosynthesis of endogenous gibberellins.

[0042] Prohexadione calcium: is a new type of plant growth regulator, belonging to the cyclohexanecarboxylic acid compound. Its chemical name is 3,5-dioxo-4-propionylcyclohexanecarboxylic acid calcium, and its chemical formula is C 20 H 22 CaO 10 It can inhibit the biosynthesis of gibberellins in plants, reduce the gibberellins content in plants, and thus delay and control the excessive growth of plants.

[0043] Methylpiperidinium chloride: a plant growth regulator, its chemical name is N,N-dimethylpiperidinium chloride, and its molecular formula is C7H 16 NCl. It has the functions of regulating plant growth, promoting reproductive growth, and inhibiting excessive stem and leaf growth, and is widely used in various crops such as cotton, wheat, and rice.

[0044] Glycyrrhizic acid: Glycyrrhizic acid has a variety of functions, including anti-inflammatory, detoxification, energy replenishment, and mood balance. Glycyrrhizic acid can also promote the normal functioning of the immune system, reduce stress and anxiety, and enhance memory.

[0045] Glycyrrhetinic acid: A large number of studies have shown that glycyrrhetinic acid has multiple biological activities, such as antiviral, anti-inflammatory, anti-ulcer, anti-tumor, hypoglycemic, and lipid-regulating.

[0046] Licoricein: Licoricein has anti-inflammatory, anti-cancer, anti-diabetic, antioxidant and liver protection properties against heavy metal toxicity.

[0047] Isoliquiritigenin: Isoliquiritigenin has anti-tumor, antioxidant, and anti-inflammatory pharmacological effects.

[0048] Seedling stage: During the seedling stage, the above-ground part of licorice gradually grows, stems and leaves begin to form, and the root system also begins to gradually expand. This stage is a critical period for the growth of licorice. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 The results of controlling the growth rate of different concentrations of uniconazole, paclobutrazol, chlormequat, prohexadione-calcium, and mepiquat are shown. *P < 0.05; ** represents P < 0.01; *** represents P < 0.001.

[0050] Figure 2 Shown is the flowering of Glycyrrhiza uralensis after paclobutrazol treatment.

[0051] Figure 3 The effect of different concentrations of uniconazole on the leaf size of licorice is shown. From left to right: control group, uniconazole 250mg / L, 150mg / L, and 80mg / L treatments.

[0052] Figure 4 The figure shows the changes in chlorophyll content in licorice leaves after treatment with different concentrations of uniconazole.

[0053] Figure 5 The effect of different concentrations of paclobutrazol on the leaf size of licorice is shown. From left to right, they are 250 mg / L, 150 mg / L, and 80 mg / L paclobutrazol.

[0054] Figure 6 The figure shows the changes in chlorophyll content in licorice leaves after treatment with different concentrations of paclobutrazol.

[0055] Figure 7 The figure shows the changes in root morphology of licorice after treatment with different concentrations of uniconazole. From left to right: control group, uniconazole 250mg / L, 150mg / L, and 80mg / L.

[0056] Figure 8 The figure shows the changes in the diameter of the rhizome of licorice after treatment with different concentrations of uniconazole.

[0057] Figure 9 The figure shows the changes in plant height of licorice after treatment with different concentrations of uniconazole. From left to right, the plants are treated with 250mg / L, 150mg / L, and 80mg / L uniconazole (the left side of each group is the control group, and the right side is the treatment group).

[0058] Figure 10The bar graph shows the changes in plant height of licorice after treatment with different concentrations of uniconazole. Note: The first measurement is before spraying, the second measurement is after one spraying, and the third measurement is after two sprayings / before sampling.

[0059] Figure 11 The figure shows the changes in underground biomass (fresh weight or wet weight) of licorice after treatment with different concentrations of uniconazole.

[0060] Figure 12 The figure shows the changes in total flavonoids content in licorice after treatment with different concentrations of uniconazole.

[0061] Figure 13 The graph shows the changes in the content of the main active ingredients in the underground part of licorice after treatment with different concentrations of uniconazole. DETAILED DESCRIPTION

[0062] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments and with reference to the accompanying drawings.

[0063] Example 1 Height Control of Different Concentrations of Uniconazole, Paclobutrazol, Chlormequat, Prohexadione-Calcium, and Mepiquat

[0064] result

[0065] Five growth regulators commonly used on the market (equinoxaline, paclobutrazol, chlormequat chloride, prohexadione-calcium, and mepiquat) were selected, and three concentrations of each were selected for field experiments. Licorice (Ural Glycyrrhiza uralensis) plants with a length of 5m and a width of 2.2m and the same growth trend were selected for spraying in each treatment group. The spraying time was in the morning before the sun rose and the licorice leaves were evenly sprayed. The spraying was done once every 10 days, for a total of two sprays. At the same time, clean water was sprayed as a control group. Samples were collected 10 days after the spraying was completed. The samples were marked and placed in an oven for drying before the active ingredients of the licorice were determined.

[0066] Planting and application time

[0067] The most suitable time for licorice planting is March to April each year. In this experiment, growth regulators were applied in mid-July (this period is the seedling stage of licorice), applied once every 10 days, and samples were taken in mid-to-early August. The samples were used for subsequent measurements of various indicators.

[0068] Application varieties and treatment concentrations

[0069] Uniconazole (5%): 250mg / L, 150mg / L, 80mg / L (Anhui Jinghong Plant Protection Co., Ltd., registration number: PD20083528)

[0070] Paclobutrazol (15%): 250 mg / L, 150 mg / L, 80 mg / L (Sichuan Runer Technology Co., Ltd., registration number: PD20081543)

[0071] Chlormequat (50%): 6000 (dilution factor), 10000 (dilution factor), 15000 (dilution factor) (Huanghua Hongcheng Co., Ltd., registration number: PD86123-2)

[0072] Prohexadione calcium (10%): 15ml / mu, 30ml / mu, 45ml / mu (Shantou Hongguang Chemical Co., Ltd., Guangdong Province, registration number: PD20220032)

[0073] Mepiquat (98%): 5g / mu, 10g / mu, 15g / mu (Nantong Jinling Agrochemical Co., Ltd., registration number: PD20096466)

[0074] Uniconazole:

[0075] The plant height of licorice showed significant changes after two sprays of uniconazole. After the first spraying, the plant height of the 250 mg / L, 150 mg / L, and 80 mg / L treatment groups decreased by 22.47, 17.22, and 12.13 cm respectively compared with the control group; after the second spraying, the plant height of the 250 mg / L, 150 mg / L, and 80 mg / L treatment groups decreased by 44.3, 30.27, and 30.36 cm respectively compared with the control group; among them, the plant height of the 250 mg / L treatment group decreased most significantly.

[0076] Paclobutrazol:

[0077] The plant height of licorice showed significant changes after two sprays of paclobutrazol. After the first spraying, the plant height of the 250 mg / L, 150 mg / L, and 80 mg / L treatment groups decreased by 25.66, 15.44, and 16.72 cm respectively compared with the control group; after the second spraying, the plant height of the 250 mg / L, 150 mg / L, and 80 mg / L treatment groups decreased by 42.39, 39.78, and 30.86 cm respectively compared with the control group; among them, the plant height of the 250 mg / L treatment group decreased most significantly.

[0078] Chlormequat:

[0079] After spraying chlormequat once, only the 6000-fold dilution treatment group showed significant changes in plant height, which was 10.02 cm lower than the control group. After the second spraying, except for the 6000-fold dilution treatment group, there was no significant change in plant height. The 15000-fold and 10000-fold dilution treatment groups both showed significant changes, which were 12.28 and 15.22 cm lower than the control group, respectively.

[0080] Prohexadione calcium:

[0081] There was no significant change in the plant height of licorice in the three treatment groups of prohexadione calcium after spraying once; after spraying twice, only the 15ml / mu treatment group showed a significant change, which was 12.26 cm lower than the control group.

[0082] Mepiquat:

[0083] There was no significant change in the plant height of licorice in the three treatment groups after spraying once with mepiquat. However, there were significant changes in the 5g / mu, 10g / mu and 15g / mu treatment groups after spraying twice, which were 11.24, 8.69 and 9.74 cm lower than the control group, respectively.

[0084] In summary, among the 15 treatment groups of 5 chemical control agents, paclobutrazol and uniconazole had relatively good effects on controlling licorice growth, among which the paclobutrazol 250 mg / L and uniconazole 250 mg / L treatment groups had the most significant effects.

[0085] According to the results, the most effective treatments for controlling licorice height were paclobutrazol 80mg / L, 150mg / L, 250mg / L and uniclobutrazol 80mg / L, 150mg / L, 250mg / L. However, the three treatment concentrations of paclobutrazol caused licorice flowering failure ( Figure 2 ), so according to all the results, uniconazole is the best for controlling the temperature, so the results of the three treatment concentrations of uniconazole were analyzed.

[0086] Example 2 Effect of Leaf Changes and Chlorophyll Content

[0087] Chlorophyll content measurement method

[0088] Measurement method: Measured using a spectrophotometer.

[0089] Ten days after the second spraying, licorice leaves were sampled (sampled from the same location on the compound leaves of each licorice plant). 0.1 g of fresh leaf sample was weighed, chopped, and placed in a 50 mL Erlenmeyer flask. 10 mL of 95% ethanol was added and the sample was allowed to stand in the dark for 3–5 hours until the leaves were completely decolorized. The extract was then filtered into a 25 mL brown volumetric flask, brought to volume with 95% ethanol, and mixed thoroughly. Using 95% ethanol as a blank control, the extract was injected into a cuvette, and absorbance was measured at 649 nm and 665 nm, respectively. Chlorophyll a and chlorophyll b concentrations (mg / L) were calculated according to the following formula; their sum was the total chlorophyll concentration (mg / g).

[0090] Ca=13.95A 665 -6.8A 649

[0091] Cb=24.96A 649 -7.32A 665

[0092] Chlorophyll content (mg / g) = (C*V*N) / (W*1000)

[0093] Where: C is the pigment concentration (mg / L); V is the volume of the extract (mL); N is the dilution factor; W is the fresh weight or dry weight of the sample (g); 1000 means 1 L = 1000 mL.

[0094] from Figure 3 It can be seen that the three concentrations of uniconazole treatment have a significant effect on the leaf size of licorice, among which the strongest effect is uniconazole 250mg / L. In addition, as the spraying concentration increases, the leaf area becomes smaller and the color becomes darker. After uniconazole treatment, the chlorophyll content of licorice increased significantly, which was 1.11 times (250mg / L uniconazole), 1.13 times (150mg / L uniconazole), and 1.12 times (80mg / L uniconazole) of the control group, and the chlorophyll content reached the maximum after the 150mg / L treatment, which was 2.503mg / g ( Figure 4 The results showed that the chlorophyll content of licorice was correlated with its leaf color, and that paclobutrazol and uniconazole treatments had a significant effect on the leaf size and chlorophyll content of licorice.

[0095] The chlorophyll content of licorice increased significantly after paclobutrazol treatment, which was 1.19 times (80 mg / L paclobutrazol), 1.11 times (150 mg / L paclobutrazol), and 1.25 times (250 mg / L paclobutrazol) of the control group, and the chlorophyll content reached the maximum after 250 mg / L treatment, which was 2.758 mg / g ( Figure 6 ).

[0096] Example 3 Root shape changes

[0097] Compared with the control group, the root morphology of the three uniconazole treatment groups was significantly thickened ( Figure 7 ), and the diameters of the reed heads increased in all three treatment concentrations compared to the control group, and there was no significant difference between the three treatment concentrations ( Figure 8 ).

[0098] Example 4 Changes in Whole Plant and Plant Height

[0099] The results show that ( Figure 9 ), the plant height of the three different concentrations of uniconazole treatment groups decreased significantly compared with the control group, among which the 250 mg / L uniconazole treatment had the most significant effect. As the concentration increased, the height control effect became more obvious, and the licorice plant height decreased significantly. According to the measurement results, spraying three different concentrations of uniconazole reduced the plant height to varying degrees. There was a significant height control effect at the first spraying, and the effect was better after the second spraying. Moreover, after both sprayings, the 250 mg / L concentration had the best height control effect ( Figure 10After spraying once, the plant height of licorice in the 250mg / L, 150mg / L, and 80mg / L treatment groups decreased by 25.53%, 21.00%, and 14.79% respectively compared with the control group; after spraying twice, the plant height of licorice in the 250mg / L, 150mg / L, and 80mg / L treatment groups decreased by 40.71%, 27.81%, and 27.90% respectively compared with the control group.

[0100] Example 5 Changes in underground biomass (fresh weight)

[0101] Among the three treatment concentrations of uniconazole, the underground biomass of 250mg / L and 150mg / L increased to varying degrees compared with the control group, with 150mg / L increasing the most, while the underground biomass of 80mg / L decreased compared with the control group ( Figure 11 The results showed that the 250mg / L and 150mg / L concentrations increased licorice biomass, potentially contributing to increased licorice yield. The 250mg / L and 150mg / L uniconazole treatments increased licorice biomass by 13.87% and 58.96%, respectively, compared to the control group.

[0102] Example 6 Effect of Uniconazole Treatment on Active Medicinal Ingredients of Licorice

[0103] Utilize UPLC-MS to measure active pharmaceutical ingredients.

[0104] Extraction Method: Grind dried licorice roots thoroughly into a powder using a grinder. Accurately weigh 0.5 g of the powder into a 10 mL centrifuge tube for extraction of the active pharmaceutical ingredient. Add 5 mL of methanol solution, ultrasonically extract for 1 hour, and centrifuge at 5000 rpm for 10 minutes in a high-speed refrigerated centrifuge. Transfer the supernatant to a new 10 mL centrifuge tube, add 5 mL of methanol solution to the remaining pellet, ultrasonicate for another 1 hour, repeat the previous step, transfer the supernatant to a 10 mL centrifuge tube, and finally dilute to 10 mL with methanol solution. Three replicates were performed for each treatment group.

[0105] Measurement conditions:

[0106] 1. Chromatographic conditions

[0107] Waters ACQUITY UPLC BEH C18 column (50 mm × 2.1 mm, 1.7 μm); flow rate: 0.3 mL / min; injection volume: 1 μL; column temperature: 35°C.

[0108] 1.1 Chromatographic conditions for high-content chemical components of licorice

[0109] Table 1 Gradient elution program of mobile phase A (0.1% formic acid-water) and B (acetonitrile)

[0110]

[0111] 1.2 Chromatographic conditions for low-content chemical components of licorice

[0112] Table 2 Gradient elution program of mobile phase A (0.1% formic acid-water) and B (acetonitrile)

[0113]

[0114] 2. Mass spectrometry conditions

[0115] The ion source was electrospray ionization (ESI), and the content was determined in multiple reaction monitoring (MRM) mode with a desolvation gas temperature of 450°C, a source temperature of 150°C, a desolvation gas flow rate of 800 L / h, a cone gas flow rate of 150 L / h, and a capillary voltage of 3000 V. The product ions of BZA were obtained by optimizing the collision-induced dissociation of its parent ion. The ion pairs, fragmentation voltage, and collision energy used for quantitative analysis are shown in Table 3.

[0116] Table 3 Mass spectrometry parameters of quantitative components

[0117]

[0118] * indicates quantification ion

[0119] Measurement method of total flavonoids content

[0120] Extraction method:

[0121] Accurately weigh 0.3 g of licorice root powder into a 50 mL centrifuge tube for total flavonoid extraction. Add 20 mL of 60% ethanol to the sample, sonicate for 30 minutes, and then centrifuge at 5000 rpm for 10 minutes in a high-speed refrigerated centrifuge. Transfer the supernatant to a new 50 mL centrifuge tube, and add another 20 mL of 60% ethanol to the pellet. Repeat the sonication and centrifugation steps, combine the supernatants, and finally bring the volume to 50 mL with 60% ethanol. Three replicates were performed for each treatment group.

[0122] Measurement method: Measured using a spectrophotometer

[0123] The total flavonoid content in licorice was determined using the KOH colorimetric method. Using a pipette, accurately transfer 0.5 mL of the sample solution from each treatment group to a 10 mL centrifuge tube. Add 0.5 mL of a 10 g / 100 mL KOH solution, incubate at room temperature for 5 minutes, and then dilute to 10 mL with methanol. Separately, take 0.5 mL of the sample solution from each treatment group and dilute to 10 mL with methanol as a blank control. Measure the absorbance at 407 nm. Calculate the total flavonoid content using the following formula.

[0124] Total flavonoid content (mg / g) = (M×V1×n×100) / (m×V2×1000)

[0125] Where: M is the regression amount (mg); V1 is the volume of the extract (mL); n is the dilution factor; m is the sample weight (g); V2 is the volume of the measured extract (mL).

[0126] The three treatment groups of uniconazole had an effect on the effective components of licorice ( Figure 13 ), compared with the control group, the 250 mg / L treatment group led to an increase in the contents of glycyrrhizic acid, glycyrrhizin, isoliquiritigenin, and glycyrrhetinic acid,

[0127] The 150mg / L treatment group resulted in an increase in the contents of glycyrrhizic acid, glycyrrhizin, and isoliquiritigenin, which were respectively increased by 43.93%, 113.73%, 83.36%, and 33.33% compared with the control group; the 150mg / L treatment group resulted in an increase in the contents of glycyrrhizic acid, glycyrrhizin, and isoliquiritigenin, which were respectively increased by 115.03%, 110.06%, and 42.88% compared with the control group; the 80mg / L treatment group resulted in an increase in the contents of glycyrrhizic acid, glycyrrhizin, and isoliquiritigenin, which were respectively increased by 24.79%, 58.26%, and 34.93% compared with the control group. In addition, from the measurement results of total flavonoids ( Figure 12 ), 250mg / L, 150mg / L, and 80mg / L treatment concentrations all significantly increased the total flavonoid content in licorice roots, increasing by 19.93%, 30.11%, and 20.36%, respectively, compared to the control group. The main medicinal part of licorice is the root, and the content of medicinally active ingredients in the root is mainly divided into flavonoids, glycyrrhizic acid, and chalcone A. Among them, flavonoids are a large class of components with strong biological activity in licorice, mainly including liquiritin, isoliquiritin, liquiritigenin, and isoliquiritigenin. The results showed that the 250mg / L treatment group significantly increased the accumulation of medicinally active ingredients such as flavonoids and glycyrrhizic acid in the underground parts of licorice.

[0128] The effects of applying five growth regulators—uniconazole, paclobutrazol, chlormequat, prohexadione-calcium, and mepiquat—were significant. Uniconazole and paclobutrazol significantly controlled licorice plant height, with plant height reductions of 44.3, 30.27, and 30.36 cm after application at 250 mg / L, 150 mg / L, and 80 mg / L, respectively. Paclobutrazol, on the other hand, reduced plant height by 42.39, 39.78, and 30.86 cm after application at 250 mg / L, 150 mg / L, and 80 mg / L, respectively. However, the results showed that all three paclobutrazol concentrations resulted in flower abortion, so uniconazole was selected as the chemical control agent for licorice. In addition, treatment with 250mg / L, 150mg / L, and 80mg / L of uniconazole increased the diameter of the rhizome of licorice. Chlorophyll content also increased significantly, suggesting that uniconazole treatment can increase the photosynthetic rate of licorice. Biomass increased to varying degrees at all treatments except 80mg / L. Determination of active ingredients in licorice found that the 250mg / L treatment significantly increased the accumulation of flavonoids and glycyrrhizic acid in the underground parts of the licorice. For licorice plants, the main medicinal parts are the roots and rhizomes, and increased accumulation of active ingredients helps improve the quality of licorice.

[0129] Although 250mg / L, 150mg / L, and 80mg / L of uniconazole showed good height control in licorice, a comprehensive analysis of the effects of the three concentrations on licorice's agronomic traits, quality, and yield ultimately found that 250mg / L, while controlling licorice plant height, could also increase the content of active ingredients such as flavonoids and glycyrrhizic acid in licorice, and could also increase licorice biomass and chlorophyll content. This provides ideas and methods for effectively controlling licorice plant height, improving licorice's lodging resistance, and enhancing the active ingredients in licorice cultivation, and lays the foundation for the subsequent cultivation of high-quality licorice medicinal materials. In summary, it was determined that spraying uniconazole at 250mg / L is the most effective method for height control in licorice cultivation.

[0130] The specific embodiments described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above are only specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. Licorice chemical control agent, characterized in that, The chemical control agent consists of uniconazole.

2. A chemical control agent for improving the lodging resistance of licorice, characterized in that: The chemical control agent consists of uniconazole.

3. Use of uniconazole in the preparation of a licorice chemical control agent, optionally, the chemical control agent has one or more of the following uses: a. Reduce the leaf area of licorice; b. Increase the chlorophyll content of licorice leaves; c. Make the roots of licorice thicker; d. Reduce the height of licorice plants; e. Increase the underground biomass of licorice; f. Increase the total flavonoid content in licorice root; g. Increase the content of one or more of glycyrrhizic acid, glycyrrhizin, isoliquiritigenin and glycyrrhetinic acid in licorice root.

4. The chemical control method of liquorice, characterized in that: The method comprises applying uniconazole to licorice, and the chemical control regulation can achieve one or more of the following uses: a. Reduce the leaf area of licorice; b. Increase the chlorophyll content of licorice leaves; c. Make the roots of licorice thicker; d. Reduce the height of licorice plants; e. Increase the underground biomass of licorice; f. Increase the total flavonoid content in licorice root; g. Increase the content of one or more of glycyrrhizic acid, glycyrrhizin, isoliquiritigenin and glycyrrhetinic acid in licorice root.

5. A method for improving the lodging resistance of licorice, characterized in that: The method comprises administering uniconazole to licorice.

6. A method for increasing the underground biomass of licorice, characterized in that: The method comprises administering uniconazole to licorice.

7. A method for increasing the accumulation of active medicinal ingredients in licorice, characterized in that: The method comprises administering uniconazole to licorice, and optionally, the pharmaceutically active ingredient comprises one or more of glycyrrhizic acid, glycyrrhizin, isoliquiritigenin and glycyrrhetinic acid.

8. The chemical control agent according to claim 1 or 2, the use according to claim 3, and the method according to any one of claims 4 to 7, characterized in that: The amount of uniconazole is 80 mg / L to 250 mg / L. Optionally, the amount of uniconazole is 80 mg / L to 240 mg / L, 80 mg / L to 230 mg / L, 80 mg / L to 220 mg / L, 80 mg / L to 210 mg / L, 80 mg / L to 200 mg / L, 80 mg / L to 190 mg / L, 80 mg / L to 180 mg / L, 80 mg / L to 170 mg / L, 80 mg / L to 160 mg / L, 80 mg / L to 150 mg / L, 80 mg / L to 140 mg / L, 80 mg / L to 130 mg / L, 80 mg / L to 120 mg / L, 80 mg / L to 110 mg / L, 80 mg / L to 100 mg / L, 80 mg / L to 90 mg / L, 90 mg / L to 250mg / L, 90 mg / L to 240mg / L, 90mg / L to 230mg / L, 90mg / L to 220mg / L, 90mg / L to 210mg / L, 90mg / L to 200mg / L, 90mg / L to 190mg / L, 90mg / L to 180mg / L, 90mg / L to 170mg / L, 90mg / L to 160mg / L, 90mg / L to 150mg / L, 90mg / L to 140mg / L, 90mg / L to 130mg / L, 90mg / L to 120mg / L, 90mg / L to 110mg / L, 90mg / L to 100mg / L, 100 100mg / L to 150mg / L, 100mg / L to 140mg / L, 100mg / L to 130mg / L, 100mg / L to 120mg / L, 100mg / L to 210mg / L, 100mg / L to 200mg / L, 100mg / L to 190mg / L, 100mg / L to 180mg / L, 100mg / L to 170mg / L, 100mg / L to 160mg / L, 100mg / L to 150mg / L, 100mg / L to 140mg / L, 100mg / L to 130mg / L, 100mg / L to 120mg / L, 100mg / L to 110mg / L, 110 mg / L to 250mg / L, 110mg / L to 240mg / L, 110mg / L to 230mg / L, 110mg / L to 220mg / L, 110mg / L to 210mg / L, 110mg / L to 200mg / L, 110mg / L to 190mg / L, 110mg / L to 180mg / L, 110mg / L to 170mg / L, 110mg / L to 160mg / L, 110mg / L to 150mg / L, 110mg / L to 140mg / L, 110mg / L to 130mg / L, 110mg / L to 120mg / L, 120mg / L to 250mg / L, 120mg / L to 240mg / L, 120mg / L to 230mg / L, 120mg / L to 220mg / L, 120mg / L to 210mg / L, 120mg / L to 200mg / L, 120mg / L to 190mg / L, 120mg / L to 180mg / L, 120mg / L to 170mg / L, 120mg / L to 160mg / L, 120mg / L to 150mg / L, 120mg / L to 140mg / L, 120mg / L to 130mg / L, 130 mg / L to 250mg / L, 130mg / L to 240mg / L, 130mg / L to 230mg / L, 130mg / L to 220mg / L, 130mg / L to 210mg / L, 130mg / L to 200mg / L, 130mg / L to 190mg / L, 130mg / L to 180mg / L, 130mg / L to 170mg / L, 130mg / L to 160mg / L, 130mg / L to 150mg / L, 130mg / L to 140mg / L, 140 mg / L to 250mg / L, 140mg / L to 240mg / L, 140mg / L to 230mg / L, 140mg / L to 220mg / L, 140mg / L to 210mg / L, 140mg / L to 200mg / L, 140mg / L to 190mg / L, 140mg / L to 180mg / L, 140mg / L to 170mg / L, 140mg / L to 160mg / L, 140mg / L to 150mg / L, 150 mg / L to 250mg / L, 150mg / L to 240mg / L, 150mg / L to 230mg / L, 150mg / L to 220mg / L, 150mg / L to 210mg / L, 150mg / L to 200mg / L, 150mg / L to 190mg / L, 150mg / L to 180mg / L, 150mg / L to 170mg / L, 150mg / L to 160 mg / L, 160mg / L to 250mg / L, 160mg / L to 240mg / L, 160mg / L to 230mg / L, 160mg / L to 220mg / L, 160mg / L to 210mg / L, 160mg / L to 200mg / L, 160mg / L to 190mg / L, 160mg / L to 180mg / L, 160mg / L to 170mg / L, 170mg / L to 250mg / L, 170mg / L to 240mg / L, 170mg / L to 230mg / L, 170mg / L to 220mg / L, 170mg / L to 210mg / L, 170mg / L to 200mg / L, 170mg / L to 190mg / L, 170mg / L to 180mg / L, 180 mg / L to 250mg / L, 180mg / L to 240mg / L, 180mg / L to 230mg / L, 180mg / L to 220mg / L, 180mg / L to 210mg / L, 180mg / L to 200mg / L, 180mg / L to 190mg / L, 190 mg / L to 250mg / L, 190mg / L to 240mg / L, 190mg / L to 230mg / L, 190mg / L to 220mg / L, 190mg / L to 210mg / L, 190mg / L to 200mg / L, 200mg / L to 250mg / L, 200mg / L to 240mg / L, 200mg / L to 230mg / L, 200mg / L to 220mg / L, 200mg / L to 210mg / L, 210mg / L to 250mg / L, 210mg / L to 240mg / L, 210mg / L to 230mg / L, 210mg / L to 220mg / L, 220 The dosage of uniconazole is 80 mg / L, 90 mg / L, 100 mg / L, 110 mg / L, 120 mg / L, 130 mg / L, 140 mg / L, 150 mg / L, 160 mg / L, 170 mg / L, 180 mg / L, 190 mg / L, 200 mg / L, 210 mg / L, 220 mg / L, 230 mg / L, 240 mg / L or 250 mg / L. Preferably, the dosage of uniconazole is 250 mg / L.

9. The chemical control agent according to claim 1, 2 or 8, the use according to claim 3 or 8, and the method according to any one of claims 4 to 8, characterized in that: The licorice is Ural licorice.

10. The chemical control agent according to claim 1, 2, 8 or 9, the use according to claim 3, 8 or 9, or the method according to any one of claims 4 to 9, characterized in that: The uniconazole is applied by spraying on the licorice leaves. Optionally, the uniconazole is applied by spraying on the licorice leaves in the morning before the sun rises. Optionally, the uniconazole is applied by spraying on the licorice leaves in the morning before the sun rises during the licorice seedling stage.

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