Methods to increase the yield of Uncaria rhynchophylla stems and promote alkaloid accumulation

Abstract

This invention discloses a method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation, which relates to the field of Uncaria rhynchophylla cultivation technology, including the following steps: (1) In February-March, during the sprouting period of new shoots of the perennial Uncaria rhynchophylla, mix phosphorus- and potassium-releasing compound slow-release microbial fertilizer with 1-3 times the amount of well-rotted farmyard manure, open a crescent-shaped shallow trench 50cm-75cm away from the plant, apply it all at once, cover with soil, and cover with ground cover or straw; (2) When the new shoots grow to 19-21cm, leave 2-7 strong buds as main branches, and when the main branches grow 6-7 pairs of lateral branches, spray the plant with the Uncaria rhynchophylla quality-inducing agent aqueous solution for the first time; (3) When the main branches grow to 15-17 pairs of lateral branches, pinch off the top, and spray the plant with the Uncaria rhynchophylla quality-inducing agent aqueous solution for the second time; (5) Harvest the 1-6 pairs of lateral branches at the base for medicinal materials in June-July, and harvest the remaining lateral branches in mid-to-late November. The method provided by this invention increases the yield of Uncaria rhynchophylla stems and promotes the accumulation of alkaloids.

Description

Technical Field

[0001] This invention relates to the field of Uncaria rhynchophylla cultivation technology, and in particular to a method for increasing the yield of Uncaria rhynchophylla stems and promoting the accumulation of alkaloids. Background Technology

[0002] Uncaria rhynchophylla (Miq.) Miq. ex Havil. is a perennial woody vine belonging to the genus Uncaria in the family Rubiaceae. It is mainly produced in Guizhou, Guangxi, Hunan, Fujian, and other provinces and autonomous regions. The main medicinal part is the hooked stem and branches. It has the effects of clearing heat and calming the liver, relieving wind and calming convulsions, lowering blood pressure, and anticonvulsant. Modern pharmaceutical and pharmacological studies have shown that alkaloids are important active ingredients in Uncaria rhynchophylla, collectively known as total alkaloids of Uncaria rhynchophylla; among them, rhynchophylline (RIN) and isorhynchophylline (IRN) are present in high amounts and have good medicinal value, playing a role in lowering blood pressure and dilating peripheral blood vessels. Uncaria rhynchophylla has been widely used in traditional Chinese medicine for infantile convulsions and cardiovascular and cerebrovascular diseases. In recent years, with the increase in the incidence of these diseases, the application and demand of Uncaria rhynchophylla in the market have increased significantly. Wild resources are decreasing, raw material prices are rising steadily, and artificial cultivation is emerging.

[0003] However, in the production of Uncaria rhynchophylla, the problems of low yield of hooked stems and branches, inconsistent alkaloid content, and high labor input costs seriously affect the enthusiasm of growers and hinder the development of the Uncaria rhynchophylla industry. The applicant's years of investigation and cultivation research have found that the following conditions will cause low yield of hooked stems and branches and low alkaloid content: (1) The internodes of the main branches of the plant are long and the number of lateral branches with hooked stems is small; (2) The internodes of the lateral branches with hooked stems are long, the hooked stem density is low, the stems are thin and the yield is low; (3) The lower part (nodes 1-6) has low yield and low content, and is dried up and discarded by the time of harvest in November; (4) After the main branches are topped, ineffective buds sprout at the nodes of each pair of lateral branches, consuming nutrients, and removing them increases labor costs; (5) Uncaria rhynchophylla only uses hooked stems and branches for medicine, and the tall main branches and lush leaves are discarded, resulting in low utilization rate; (6) The alkaloid content of hooked stems and branches is unstable, affecting the therapeutic effect. Improving quality and increasing income are the bottleneck issues that need to be addressed urgently in the production of Uncaria rhynchophylla. Summary of the Invention

[0004] To address the problems existing in the prior art, this invention provides a method for increasing the yield of Uncaria rhynchophylla hook stems and promoting alkaloid accumulation, which can improve the yield of Uncaria rhynchophylla hook stems and branches and the alkaloid content.

[0005] To achieve the above-mentioned objectives, the technical solution of the present invention is as follows:

[0006] A method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation includes the following steps:

[0007] (1) In February-March, during the sprouting period of new shoots of Uncaria rhynchophylla, mix phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer with 1-3 times the amount of well-rotted farmyard manure, open a crescent-shaped shallow trench 50cm-75cm away from the plant, apply it all at once, cover with soil, and cover with ground cover or straw.

[0008] (2) When the new shoots grow to 19-21cm, leave 2-7 strong shoots as main branches. When the main branches grow 6-7 pairs of lateral branches, spray the plant with the first aqueous solution of Uncaria rhynchophylla quality-inducing agent.

[0009] (3) When the main branch grows to 15-17 pairs of lateral branches, pinch off the top and spray the plant with a second aqueous solution of Uncaria rhynchophylla quality-inducing agent.

[0010] (4) Harvest the 1-6 pairs of lateral branches at the base in June-July for medicinal purposes, and harvest the remaining lateral branches in mid-to-late November;

[0011] The phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer, by weight, comprises: 15-25 parts of Uncaria rhynchophylla stems and leaves (3:1 weight ratio), 6-12 parts of Luo Han Guo (monk fruit) extraction waste or residual fruit, 5-11 parts of tung oil bran, 5-11 parts of earthworm excrement, 5-11 parts of calcium magnesium phosphate fertilizer, 4-10 parts of silkworm residue, 4-10 parts of soybean meal, 3-9 parts of biochar, 2-4 parts of sugarcane filter mud, 3-9 parts of potassium humate, 0.1-0.3 parts of Pseudomonas fluorescens, and Bacillus mucilaginosus.

[0012] 0.09-0.11 parts, 0.3-0.5 parts of phosphate-solubilizing bacteria, 0.1-0.3 parts of *Vibrio butyricum* lysozyme, 0.1-0.3 parts of *Aspergillus niger*, 0.1-0.3 parts of yeast, and 5-11 parts of slow-release compound fertilizer; wherein the effective viable count of *Pseudomonas fluorescens* is ≥200 million / ml, the effective viable count of *Bacillus mucilaginosus* is ≥5 billion / g, and the effective viable count of the phosphate-solubilizing bacteria is ≥1.0

[0013] ×10cfu / g, the N:P:K ratio in the slow-release compound fertilizer is 15:8:18;

[0014] Each kilogram of the above-mentioned Uncaria rhynchophylla extract inducing aqueous solution comprises: 5-7 mg of strigolactone, 4-6 mg of folinic acid, 16-24 mg of benzylaminopurine, 8-12 mg of biogenic organic vanadium, 10-15 mg of amino acid zinc, 16-24 mg of amino acid chelated boron, 81-99 mg of organosilicon, 89-111 mg of amino acid chelated rare earth, 800-1200 mg of sugar alcohol calcium magnesium, and 800-1200 mg of potassium dihydrogen phosphate.

[0015] Furthermore, the phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer, by weight, comprises: 20 parts of Uncaria rhynchophylla stems and leaves in a weight ratio of 3:1, 8.7 parts of Luo Han Guo extraction waste or residual fruit, 8 parts of tung oil bran, 8 parts of earthworm excrement, 8 parts of calcium magnesium phosphate fertilizer, 7 parts of silkworm residue, 7 parts of soybean meal, 6 parts of biochar, 3 parts of sucrose filter mud, 6 parts of potassium humate, 0.2 parts of Pseudomonas fluorescens, 0.1 parts of Bacillus mucilaginosus, 0.4 parts of phosphate-solubilizing bacteria, 0.2 parts of Vibrio butyricum fibrosum, 0.2 parts of Aspergillus niger, 0.2 parts of yeast, and 8 parts of slow-release compound fertilizer. The effective viable count of Pseudomonas fluorescens is ≥200 million / ml, the effective viable count of Bacillus mucilaginosus is ≥5 billion / g, the effective viable count of phosphate-solubilizing bacteria is ≥1.0×10 CFU / g, and the N:P:K ratio in the slow-release compound fertilizer is 15:8:18.

[0016] Furthermore, the preparation method of the phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer is as follows:

[0017] S1. Weigh out the following ingredients according to the specified ratio: Uncaria rhynchophylla stems and leaves, Luo Han Guo extract residue or fruit residue, tung oil bran, calcium magnesium phosphate fertilizer, silkworm residue, soybean meal, and sugarcane filter mud. Mix them together. The Uncaria rhynchophylla stems and leaves are crushed, and the Luo Han Guo extract residue or fruit residue is crushed.

[0018] S2. Add Vibrio butyricum, Aspergillus niger, and yeast according to the formula, and add water to the moisture content to 39%-41%. Carry out rainproof composting and fermentation. During the fermentation process, when the temperature in the center reaches 50℃, turn the pile until it is fully decomposed.

[0019] S3. Add biochar, earthworm excrement, potassium humate, Bacillus mucilaginosus, and phosphate-solubilizing bacteria according to the formula, maintain the moisture content at 24%-26%, and carry out composting culture.

[0020] S4. Before use, add fluorescent Pseudomonas aeruginosa and slow-release compound fertilizer according to the ratio.

[0021] Furthermore, each kilogram of the above-mentioned Uncaria rhynchophylla extract inducing aqueous solution comprises: 6 mg of strigolactone, 5 mg of folinic acid, 20 mg of benzylaminopurine, 10 mg of bio-organic vanadium, 12 mg of amino acid ester, 20 mg of amino acid zinc, 90 mg of amino acid chelated boron, 100 mg of organosilicon, 1000 mg of amino acid chelated rare earth, 200 mg of sugar alcohol calcium magnesium, and 1000 mg of potassium dihydrogen phosphate.

[0022] Furthermore, the method for preparing the aqueous solution of the Uncaria rhynchophylla extract inducer is as follows:

[0023] S5. Weigh out strigolactone and benzylaminopurine according to the ratio, add acetone until completely dissolved, and then dilute with distilled water to form the first mother liquor.

[0024] S6. Weigh out the following ingredients according to the specified ratio: leaf extract, organic vanadium, amino acid lipids, zinc amino acid chelated boron, organosilicon, rare earth amino acid chelated sugar alcohol, calcium magnesium, and potassium dihydrogen phosphate. Mix them and dissolve them in distilled water to form the second mother liquor.

[0025] S7. Combine the first mother liquor and the second mother liquor, and dilute with distilled water to the required concentration to obtain an aqueous solution of Uncaria rhynchophylla extract inducing agent.

[0026] Furthermore, the method for preparing the bio-organic vanadium is as follows:

[0027] S8. Select plump soybeans, soak them in a 0.02% potassium permanganate solution for 1 minute, take them out, rinse them with distilled water to wash off the potassium permanganate on the surface of the soybeans, then soak them in 45℃ hot water for 20 minutes, take them out, drain the surface water, and set aside.

[0028] S9. Soak soybeans in a sodium metavanadate aqueous solution with a concentration of 780mg / L-820mg / L, and keep them in a constant temperature biological incubator at 40℃ for 8 hours. Take them out, place them in a germination box, spread them out to a thickness of 2-3cm, cover them, and place them in a constant temperature biological incubator at 28℃ for 72 hours in the dark. Open the lid every day and sprinkle the same concentration of sodium metavanadate aqueous solution, and roll the soybeans to keep the soybean surface moist.

[0029] S10. After the soybeans sprout, rinse the soybean sprouts with tap water, then rinse them with distilled water to wash away the sodium metavanadate on the surface of the soybean sprouts. Then dry them in a hot air drying oven at 60°C until constant weight, pulverize the soybeans, and obtain soybean tooth powder containing organic vanadium.

[0030] Furthermore, during the plant's growth period, maintain a relative soil humidity of 60-80%.

[0031] Furthermore, in step (3), 14-16 days after the second spraying of the plant with the Uncaria rhynchophylla quality-inducing agent, the plant is sprayed with the Uncaria rhynchophylla quality-inducing agent aqueous solution for the third time.

[0032] Furthermore, in step (2), 5-6 strong buds are retained as main branches.

[0033] The method provided by this invention has at least the following beneficial effects:

[0034] 1. The method provided by this invention uses a compound slow-release microbial fertilizer that solubilizes phosphorus and potassium, combined with farmyard manure as root fertilizer to improve the soil, and an aqueous solution of Uncaria rhynchophylla quality-inducing agent to regulate plant growth. This method not only promotes the synthesis and accumulation of Uncaria rhynchophylla alkaloids, but also increases the length of lateral branches while shortening the length of internodes, increasing the size of hook stems, increasing the density of hook stems, and improving the yield of hook stems. It can also inhibit the germination of ineffective buds after the main branches are removed, reducing the labor costs incurred due to bud removal.

[0035] 2. The method provided by this invention uses 5 strong buds as main branches, which can obtain more hooked stem weight.

[0036] 3. The method of this invention uses Uncaria rhynchophylla quality-inducing agent for the first time when the main branch has grown 6-7 lateral branches. This not only promotes the growth of lateral branches, but also increases the alkaloid content at this stage. Timely harvesting allows it to be used as a medicinal material, turning waste into treasure.

[0037] Other advantages, objectives and features of the present invention will become apparent in part from the following description, and in part from those skilled in the art through study and practice of the invention. Attached Figure Description

[0038] Figure 1 This is a comparison diagram of the lateral branches at the 13th node of the main branch of Uncaria rhynchophylla cultivated in Example 1 and Comparative Example 3 of the present invention (the left side is the lateral branch of Uncaria rhynchophylla in Comparative Example 3, and the right side is the lateral branch of Uncaria rhynchophylla in Example 1).

[0039] Figure 2 This is a comparison diagram of a Uncaria rhynchophylla plant (with hooked stem) cultivated in Example 1 and Comparative Example 3 of the present invention (the left side is the Uncaria rhynchophylla plant of Comparative Example 3, and the right side is the Uncaria rhynchophylla plant of Example 1). Detailed Implementation

[0040] The present invention will now be described in further detail to enable those skilled in the art to implement it based on the description.

[0041] Example 1

[0042] Methods to increase the yield of Uncaria rhynchophylla stems and promote alkaloid accumulation include the following steps:

[0043] (1) In February-March, during the sprouting period of new shoots of Uncaria rhynchophylla, mix 1 kg of phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer with 2 kg of well-rotted farmyard manure (such as manure from poultry and livestock and straw bedding material). Apply the mixture in one go by opening a crescent-shaped shallow trench 50-75 cm away from the plant, cover it with soil, and cover it with ground cover or straw.

[0044] The phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer, by weight, comprises: 20 parts of Uncaria rhynchophylla stems and leaves in a 3:1 weight ratio, 8.7 parts of Luo Han Guo extract residue, 8 parts of tung oil bran, 8 parts of earthworm excrement, 8 parts of calcium magnesium phosphate fertilizer, 7 parts of silkworm residue, 7 parts of soybean meal, 6 parts of biochar, 3 parts of sucrose filter mud, 6 parts of potassium humate, 0.2 parts of Pseudomonas fluorescens (effective viable count ≥ 200 million / ml), 0.1 parts of Bacillus mucilaginosus (effective viable count ≥ 5 billion / g), 0.4 parts of phosphate-solubilizing bacteria (effective viable count ≥ 1.0 × 10 CFU / g), 0.2 parts of Vibrio butyricum fibrolyticus, 0.2 parts of Aspergillus niger, 0.2 parts of yeast, and 8 parts of slow-release compound fertilizer (N:P:K = 15:8:18).

[0045] Its preparation method is as follows:

[0046] S1. Weigh out the following ingredients according to the specified proportions: Uncaria rhynchophylla stems and leaves, monk fruit extract residue, tung oil bran, and calcium magnesium phosphate fertilizer.

[0047] Silkworm residue, soybean meal, and sugarcane filter mud are mixed together, wherein: Uncaria rhynchophylla stems and leaves are crushed, and Luo Han Guo extraction waste is crushed;

[0048] S2. Add Vibrio butyricum, Aspergillus niger, and yeast according to the formula ratio, and add water until the moisture content is [missing information].

[0049] 39%-41%, and compost in a rain-sheltered manner. During the fermentation process, when the temperature at the center reaches 50℃, turn the pile over until it is fully decomposed.

[0050] S3. Add biochar, earthworm excrement, potassium humate, Bacillus mucilaginosus, and phosphate-solubilizing bacteria according to the formula, maintain the moisture content at 24%-26%, and carry out composting culture.

[0051] S4. Before use, add fluorescent Pseudomonas aeruginosa and slow-release compound fertilizer according to the ratio.

[0052] (2) When the new shoots grow to 19-21cm, leave 5 strong shoots as main branches. When the main branches grow 6-7 pairs of lateral branches, spray the plant with the first aqueous solution of Uncaria rhynchophylla quality-inducing agent.

[0053] The aqueous solution of the Uncaria rhynchophylla extract inducer per kilogram contains: 6 mg of strigolactone, 5 mg of folinic acid, 20 mg of benzylaminopurine, 10 mg of bio-organic vanadium, 12 mg of amino acid ester, 20 mg of amino acid zinc, 90 mg of amino acid chelated boron, 100 mg of organosilicon, 1000 mg of amino acid chelated rare earth, 200 mg of sugar alcohol calcium magnesium, and 1000 mg of potassium dihydrogen phosphate.

[0054] Its preparation method is as follows:

[0055] S5. Weigh out strigolactone and benzylaminopurine according to the ratio, add acetone until completely dissolved, and then dilute with distilled water to form the first mother liquor.

[0056] S6. Weigh out the following ingredients according to the specified ratio: leaf extract, organic vanadium, amino acid lipids, zinc amino acid chelated boron, organosilicon, rare earth amino acid chelated sugar alcohol, calcium magnesium, and potassium dihydrogen phosphate. Mix them and dissolve them in distilled water to form the second mother liquor.

[0057] S7. Combine the first mother liquor and the second mother liquor, and dilute with distilled water to the required concentration to obtain an aqueous solution of Uncaria rhynchophylla extract inducing agent.

[0058] The method for preparing the bio-organic vanadium is as follows:

[0059] S8. Select plump soybeans, soak them in a 0.02% potassium permanganate solution for 1 minute, take them out, rinse them 4 times with distilled water to wash off the potassium permanganate on the surface of the soybeans, then soak them in 45℃ hot water for 20 minutes, take them out, drain the surface water, and set them aside.

[0060] S9. Soak soybeans in an 800 mg / L sodium metavanadate aqueous solution, place them in a 40°C constant temperature biological incubator for 8 hours, remove them, place them in a germination box, spread them out to a thickness of 2-3 cm, cover them, and place them in a 28°C constant temperature biological incubator for 72 hours in the dark. Open the lid every day and sprinkle with an 800 mg / L sodium metavanadate aqueous solution. During the process of sprinkling the concentrated sodium metavanadate aqueous solution, roll the soybeans to keep the soybean surface moist.

[0061] S10. After the soybeans sprout, rinse the soybean sprouts three times with tap water, and then rinse them four times with distilled water to remove sodium metavanadate from the surface of the soybean sprouts. Then dry them in a hot air drying oven at 60°C until constant weight, crush the soybeans, and pass them through a 100-mesh standard sieve to obtain soybean tooth powder containing organic vanadium, which is biological organic vanadium.

[0062] (3) When the main branch grows to 16 pairs of lateral branches, pinch off the top and spray the plant with a second solution of Uncaria rhynchophylla quality-inducing agent.

[0063] (4) Harvest the 1-6 pairs of lateral branches at the base in June-July for medicinal purposes; harvest the 7-16 pairs of lateral branches in mid-to-late November.

[0064] Remove the leaves from the harvested lateral branches, let them air dry, and cut them into 1.5-2.0cm long hooked stems and internodes.

[0065] During the plant's growth period, maintain a relative soil humidity of 60-80%.

[0066] Example 2

[0067] Methods to increase the yield of Uncaria rhynchophylla stems and promote alkaloid accumulation include the following steps:

[0068] (1) In February-March, during the sprouting period of new shoots of Uncaria rhynchophylla, mix 1 kg of phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer with 2 kg of well-rotted farmyard manure (such as manure from poultry and livestock and straw bedding material). Apply the mixture in one go by opening a crescent-shaped shallow trench 50-75 cm away from the plant, cover it with soil, and cover it with ground cover or straw.

[0069] The phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer, by weight, comprises: 25 parts of Uncaria rhynchophylla stems and leaves in a 3:1 weight ratio, 12 parts of Luo Han Guo extract residue, 11 parts of tung oil bran, 11 parts of earthworm excrement, 11 parts of calcium magnesium phosphate fertilizer, 10 parts of silkworm residue, 10 parts of soybean meal, 9 parts of biochar, 4 parts of sucrose filter mud, 9 parts of potassium humate, 0.3 parts of *Pseudomonas fluorescens* (effective viable count ≥ 200 million / ml), 0.11 parts of *Bacillus mucilaginosus* (effective viable count ≥ 5 billion / g), 0.5 parts of phosphate-solubilizing bacteria (effective viable count ≥ 1.0 × 10 CFU / g), 0.3 parts of *Vibrio butyricum*, 0.3 parts of *Aspergillus niger*, 0.3 parts of yeast, and 11 parts of slow-release compound fertilizer (N:P:K = 15:8:18).

[0070] Its preparation method is as follows:

[0071] S1. Weigh out the following ingredients according to the specified proportions: Uncaria rhynchophylla stems and leaves, monk fruit extract residue, tung oil bran, and calcium magnesium phosphate fertilizer.

[0072] Silkworm residue, soybean meal, and sugarcane filter mud are mixed together, wherein: Uncaria rhynchophylla stems and leaves are crushed, and Luo Han Guo extraction waste is crushed;

[0073] S2. Add Vibrio butyricum, Aspergillus niger, and yeast according to the formula ratio, and add water until the moisture content is [missing information].

[0074] 39%-41%, and compost in a rain-sheltered manner. During the fermentation process, when the temperature at the center reaches 50℃, turn the pile over until it is fully decomposed.

[0075] S3. Add biochar, earthworm excrement, potassium humate, Bacillus mucilaginosus, and phosphate-solubilizing bacteria according to the formula, maintain the moisture content at 24%-26%, and carry out composting culture.

[0076] S4. Before use, add fluorescent Pseudomonas aeruginosa and slow-release compound fertilizer according to the ratio.

[0077] (2) When the new shoots grow to 19-21cm, leave 5 strong shoots as main branches. When the main branches grow 6-7 pairs of lateral branches, spray the plant with the first aqueous solution of Uncaria rhynchophylla quality-inducing agent.

[0078] The aqueous solution of the Uncaria rhynchophylla extract inducer per kilogram contains: 7 mg of strigolactone, 6 mg of folinic acid, 24 mg of benzylaminopurine, 12 mg of bio-organic vanadium, 15 mg of amino acid ester, 24 mg of amino acid zinc, 99 mg of amino acid chelated boron, 111 mg of organosilicon, 1200 mg of amino acid chelated rare earth, 220 mg of sugar alcohol calcium magnesium, and 1200 mg of potassium dihydrogen phosphate.

[0079] Its preparation method is as follows:

[0080] S5. Weigh out strigolactone and benzylaminopurine according to the ratio, add acetone until completely dissolved, and then dilute with distilled water to form the first mother liquor.

[0081] S6. Weigh out the following ingredients according to the specified ratio: leaf extract, organic vanadium, amino acid lipids, zinc amino acid chelated boron, organosilicon, rare earth amino acid chelated sugar alcohol, calcium magnesium, and potassium dihydrogen phosphate. Mix them and dissolve them in distilled water to form the second mother liquor.

[0082] S7. Combine the first mother liquor and the second mother liquor, and dilute with distilled water to the required concentration to obtain an aqueous solution of Uncaria rhynchophylla extract inducing agent.

[0083] The method for preparing the bio-organic vanadium is as follows:

[0084] S8. Select plump soybeans, soak them in a 0.02% potassium permanganate solution for 1 minute, take them out, rinse them 4 times with distilled water to wash off the potassium permanganate on the surface of the soybeans, then soak them in 45℃ hot water for 20 minutes, take them out, drain the surface water, and set them aside.

[0085] S9. Soak soybeans in an 820 mg / L sodium metavanadate aqueous solution, place them in a 40°C constant temperature biological incubator for 8 hours, remove them, place them in a germination box, spread them out to a thickness of 2-3 cm, cover them, and place them in a 28°C constant temperature biological incubator for 72 hours in the dark. Open the lid every day and sprinkle with an 820 mg / L sodium metavanadate aqueous solution. During the process of sprinkling the concentrated sodium metavanadate aqueous solution, roll the soybeans to keep the soybean surface moist.

[0086] S10. After the soybeans sprout, rinse the soybean sprouts three times with tap water, and then rinse them four times with distilled water to remove sodium metavanadate from the surface of the soybean sprouts. Then dry them in a hot air drying oven at 60°C until constant weight, crush the soybeans, and pass them through a 100-mesh standard sieve to obtain soybean tooth powder containing organic vanadium, which is biological organic vanadium.

[0087] (3) When the main branch grows to 17 pairs of lateral branches, pinch off the top and spray the plant with a second solution of Uncaria rhynchophylla quality-inducing agent.

[0088] (4) Harvest the 1-6 pairs of lateral branches at the base in June-July for medicinal purposes; harvest the 7-17 pairs of lateral branches in mid-to-late November.

[0089] Remove the leaves from the harvested lateral branches, let them air dry, and cut them into 1.5-2.0cm long hooked stems and internodes.

[0090] During the plant's growth period, maintain a relative soil humidity of 60-80%.

[0091] Example 3

[0092] Methods to increase the yield of Uncaria rhynchophylla stems and promote alkaloid accumulation include the following steps:

[0093] (1) In February-March, during the sprouting period of new shoots of Uncaria rhynchophylla, mix 1 kg of phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer with 2 kg of well-rotted farmyard manure (such as manure from poultry and livestock and straw bedding material). Apply the mixture in one go by opening a crescent-shaped shallow trench 50-75 cm away from the plant, cover it with soil, and cover it with ground cover or straw.

[0094] The phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer, by weight, comprises: 15 parts of Uncaria rhynchophylla stems and leaves in a 3:1 weight ratio, 6 parts of Siraitia grosvenorii fruit residue, 5 parts of tung oil bran, 5 parts of earthworm excrement, 5 parts of calcium magnesium phosphate fertilizer, 4 parts of silkworm residue, 4 parts of soybean meal, 3 parts of biochar, 2 parts of sucrose filter mud, 3 parts of potassium humate, 0.1 parts of Pseudomonas fluorescens (effective viable count ≥ 200 million / g), 0.09 parts of Bacillus mucilaginosus (effective viable count ≥ 5 billion / g), 0.3 parts of phosphate-solubilizing bacteria (effective viable count ≥ 1.0 × 10 CFU / g), 0.1 parts of Vibrio butyricum lysozyme, 0.1 parts of Aspergillus niger, 0.1 parts of yeast, and 5 parts of slow-release compound fertilizer (N:P:K = 15:8:18).

[0095] Its preparation method is as follows:

[0096] S1. Weigh out the following ingredients according to the specified ratio: Uncaria rhynchophylla stems and leaves, Siraitia grosvenorii fruit residue, tung oil bran, calcium magnesium phosphate fertilizer, silkworm residue, soybean meal, and sugarcane filter mud. Mix them together. The Uncaria rhynchophylla stems and leaves are crushed, and the Siraitia grosvenorii fruit residue is rolled.

[0097] S2. Add Vibrio butyricum, Aspergillus niger, and yeast according to the formula ratio, and add water until the moisture content is [missing information].

[0098] 39%-41%, and compost in a rain-sheltered manner. During the fermentation process, when the temperature at the center reaches 50℃, turn the pile over until it is fully decomposed.

[0099] S3. Add biochar, earthworm excrement, potassium humate, Bacillus mucilaginosus, and phosphate-solubilizing bacteria according to the formula, maintain the moisture content at 24%-26%, and carry out composting culture.

[0100] S4. Before use, add fluorescent Pseudomonas aeruginosa and slow-release compound fertilizer according to the ratio.

[0101] (2) When the new shoots grow to 19-21cm, leave 5 strong shoots as main branches. When the main branches grow 6-7 pairs of lateral branches, spray the plant with the first aqueous solution of Uncaria rhynchophylla quality-inducing agent.

[0102] The aqueous solution of the Uncaria rhynchophylla extract inducer per kilogram contains: 5 mg of strigolactone, 4 mg of folinic acid, 16 mg of benzylaminopurine, 8 mg of bio-organic vanadium, 10 mg of amino acid ester, 16 mg of amino acid zinc, 81 mg of amino acid chelated boron, 89 mg of organosilicon, 800 mg of amino acid chelated rare earth, 180 mg of sugar alcohol calcium magnesium, and 800 mg of potassium dihydrogen phosphate.

[0103] Its preparation method is as follows:

[0104] S5. Weigh out strigolactone and benzylaminopurine according to the ratio, add acetone until completely dissolved, and then dilute with distilled water to form the first mother liquor.

[0105] S6. Weigh out the following ingredients according to the specified ratio: leaf extract, organic vanadium, amino acid lipids, zinc amino acid chelated boron, organosilicon, rare earth amino acid chelated sugar alcohol, calcium magnesium, and potassium dihydrogen phosphate. Mix them and dissolve them in an appropriate amount of distilled water to form the second mother liquor.

[0106] S7. Combine the first mother liquor and the second mother liquor, and dilute with distilled water to the required concentration to obtain an aqueous solution of Uncaria rhynchophylla extract inducing agent.

[0107] The method for preparing the bio-organic vanadium is as follows:

[0108] S8. Select plump soybeans, soak them in a 0.02% potassium permanganate solution for 1 minute, take them out, rinse them 4 times with distilled water to wash off the potassium permanganate on the surface of the soybeans, then soak them in 45℃ hot water for 20 minutes, take them out, drain the surface water, and set them aside.

[0109] S9. Soak soybeans in a 780 mg / L sodium metavanadate aqueous solution, place them in a 40°C constant temperature biological incubator for 8 hours, remove them, place them in a germination box, spread them out to a thickness of 2-3 cm, cover them, and place them in a 28°C constant temperature biological incubator for 72 hours in the dark. Open the lid every day and sprinkle with a 780 mg / L sodium metavanadate aqueous solution. During the process of sprinkling the concentrated sodium metavanadate aqueous solution, roll the soybeans to keep the soybean surface moist.

[0110] S10. After the soybeans sprout, rinse the soybean sprouts three times with tap water, and then rinse them four times with distilled water to remove sodium metavanadate from the surface of the soybean sprouts. Then dry them in a hot air drying oven at 60°C until constant weight, crush the soybeans, and pass them through a 100-mesh standard sieve to obtain soybean tooth powder containing organic vanadium, which is biological organic vanadium.

[0111] (3) When the main branch grows to 15 pairs of lateral branches, pinch off the top and spray the plant with a second solution of Uncaria rhynchophylla quality-inducing agent.

[0112] (4) Harvest the 1-6 pairs of lateral branches at the base in June-July for medicinal purposes; harvest the 7-15 pairs of lateral branches in mid-to-late November.

[0113] Remove the leaves from the harvested lateral branches, let them air dry, and cut them into 1.5-2.0cm long hooked stems and internodes.

[0114] During the plant's growth period, maintain a relative soil humidity of 60-80%.

[0115] Example 4

[0116] Similar to Example 1, except that in step (3), 15 days after the second spraying of the Uncaria rhynchophylla quality-inducing agent on the plant, the plant is sprayed with the Uncaria rhynchophylla quality-inducing agent aqueous solution for the third time.

[0117] Example 5

[0118] Same as in Example 1, except that in step (2), two strong buds are left as main branches.

[0119] Example 6

[0120] Same as in Example 1, except that in step (2), three strong buds are left as main branches.

[0121] Example 7

[0122] Similar to Example 1, except that in step (2), four strong buds are left as main branches.

[0123] Example 8

[0124] Same as in Example 1, except that in step (2), 6 strong buds are left as main branches.

[0125] Example 9

[0126] Same as in Example 1, except that in step (2), 7 strong buds are left as main branches.

[0127] Comparative Example 1

[0128] Same as Example 1, except that gibberellin and abscisic acid are used instead of the aqueous solution of Uncaria rhynchophylla extract inducer, that is:

[0129] In step (2), when the main branch has grown 6-7 pairs of lateral branches, the plant is sprayed with gibberellin and abscisic acid for the first time.

[0130] In step (3), when the main branch grows to 16 pairs of lateral branches, the top is removed, and the plant is sprayed with gibberellin solution and abscisic acid for the second time.

[0131] The gibberellin concentration was 1300 mg / L, and the application rate was 3 ml / plant; the abscisic acid concentration was 15 mol / L, and the application rate was 7 ml / plant.

[0132] Comparative Example 2

[0133] Same as Example 1, except that the phosphorus and potassium solubilizing compound slow-release microbial fertilizer is not used, and the amount of farmyard manure is increased, that is:

[0134] In step (1), 4 kg of well-rotted farmyard manure (such as manure from poultry and livestock and straw bedding) is applied in a crescent-shaped shallow trench 50cm-75cm away from the plant, covered with soil, and then covered with ground cover or straw.

[0135] Comparative Example 3

[0136] The existing method is as follows: In February and March, during the sprouting period of new shoots of the perennial Uncaria rhynchophylla, apply 4 kg of well-rotted farmyard manure (such as manure made from poultry and livestock excrement and straw bedding) per plant, digging shallow trenches around the plant and applying it, then covering it with soil. When the new shoots reach 20 cm in length, retain 5 strong buds as main branches. In mid-April, apply 30 g of urea per plant; in late June, apply 0.5 kg of inorganic compound fertilizer (N:P:K = 15:15:15) per plant. When the main branches grow to 16 pairs of lateral branches, pinch off the top. Weed once each in May and July. From August to October, manually remove ineffective buds at irregular intervals. Harvest the lateral branches in mid-to-late November. Remove the leaves, air-dry, and cut 1.5-2.0 cm long hooked stems and internodes.

[0137] Comparative Example 4

[0138] Similar to Example 1, the difference is that in step (2), when the main branch grows 8 pairs of lateral branches, the plant is sprayed with an aqueous solution of Uncaria rhynchophylla quality-inducing agent for the first time.

[0139] Experimental Comparison 1

[0140] The comparative experiment was conducted from February 23 to December 3, 2023. Seven treatments were set up according to Examples 1, 2, 3, and 4, and the methods described in Comparative Examples 1-3 were performed, with each treatment lasting 27 minutes. 2(9 plants), 3 replicates. Regular observations were conducted, with other management practices consistent. Lateral branches from the base of the main branches (1-6 pairs) were harvested on June 28th; the remaining lateral branches were harvested on November 29th. After drying, three plants were randomly selected to measure the length of the lateral branch at the 13th node of the main branch and the internode length and stem diameter at the 5th node. The total weight of the hook stems was determined by plot, and the alkaloid (total alkaloid) content of the hook stems was detected by high-performance liquid chromatography. The results are shown in Table 1.

[0141] Table 1 Comparison of Uncaria rhynchophylla yield and total alkaloid content between the present invention and prior art

[0142]

[0143] The results showed that, compared with Comparative Example 3, Examples 1, 2, 3, and 4 (1) the present invention could significantly promote the elongation and growth of Uncaria rhynchophylla lateral branches by 45.27%-53.76%; (2) it could shorten the internode length of lateral branches by 35.27%-36.38%; (3) the stems at the same node thickened by 9.65%-21.05%; and (4) the total yield of Uncaria rhynchophylla stems in the plot was significantly higher than that of Comparative Example 3.

[0144] The yield increased by 31.30%-49.57% between 3.02kg and 3.44kg; (5) The content of rhizoma alkaloid in the rhizome was 0.5711mg / g-0.6084mg / g, which was 20.56%-28.43% higher than that of the prior art. The phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer combined with farmyard manure was used as root fertilizer to improve the soil, and the rhizoma quality-inducing agent aqueous solution was used to regulate plant growth. Compared with the prior art, the method provided by the present invention can not only promote the synthesis and accumulation of rhizoma alkaloids, but also increase the length of lateral branches while shortening the length of internodes, increasing the rhizome length and increasing the rhizome density. It can also inhibit the germination of ineffective buds after the main branch is removed, and reduce the labor cost invested by removing buds. Please refer to the comparison diagram of the rhizoma lateral branches cultivated in Example 1 and Comparative Example 3 and the comparison diagram of the obtained medicinal materials. Figure 1 , Figure 2 2. Compared with Comparative Examples 1 and 2, Examples 1, 2, and 3 simultaneously used phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer combined with farmyard manure as root fertilizer to improve the soil, and used an aqueous solution of Uncaria rhynchophylla quality-inducing agent to regulate plant growth. This significantly increased the length of lateral branches, shortened the internode length, thickened the hook stems, and increased the yield of hook stems. It also promoted the synthesis and accumulation of Uncaria rhynchophylla alkaloids, and its synergistic effect was significant.

[0145] Experimental Comparison 2

[0146] The comparative experiment was conducted from February 23 to December 3, 2023, with six treatments following the methods described in Examples 1 and 5-9, each lasting 27 minutes. 2(9 plants), 3 replicates. Regular observation and other management practices were maintained. Lateral branches from the base of the main branches (1-6 pairs) were harvested on June 28; the remaining pairs of lateral branches were harvested on November 29.

[0147] Table 2. Effects of retaining different numbers of buds as main branches on the yield of Uncaria rhynchophylla medicinal materials.

[0148]

[0149]

[0150] As shown in Table 2, retaining 5 strong buds as main branches can yield more hooked stems of Uncaria rhynchophylla medicinal material.

[0151] Experimental Comparison 3

[0152] The comparative experiment was conducted from February 23 to June 28, 2023, with one treatment following the method described in Comparative Example 4. 27m 2 Nine plants were planted in total. Management was consistent, and regular observations were conducted. Observations showed that the eighth pair of lateral branches emerged successively in late May or June. At this time, the lateral branches below the seventh pair on the Uncaria rhynchophylla plant began to wither. Although the plants were sprayed with an Uncaria rhynchophylla quality-inducing agent solution for the first time at this time, by harvest time on June 28th, the lateral branches below the seventh pair on five Uncaria rhynchophylla plants were withered, while the lateral branches below the seventh pair on four plants appeared relatively normal, but their alkaloid content was still low, at 0.3215 mg / g, 0.3125 mg / g, 0.2918 mg / g, and 0.2834 mg / g respectively. The method used in this invention involves applying the Uncaria rhynchophylla quality-inducing agent for the first time when the main branch has grown 6-7 lateral branches. This not only promotes lateral branch growth but also increases the alkaloid content at this stage. Timely harvesting allows for medicinal use, turning waste into treasure.

[0153] Although embodiments of the present invention have been disclosed above, they are not limited to the applications listed in the specification and embodiments. They can be applied to various fields suitable for the present invention. For those skilled in the art, other modifications can be easily made. Therefore, without departing from the general concept defined by the claims and their equivalents, the present invention is not limited to the specific details and illustrations shown and described herein.

Claims

1. A method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation, characterized in that, Includes the following steps: (1) In February-March, during the sprouting period of new shoots of Uncaria rhynchophylla, mix phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer with 1-3 times the amount of well-rotted farmyard manure, open a crescent-shaped shallow trench 50cm-75cm away from the plant, apply it all at once, cover with soil, and cover with ground cover or straw. (2) When the new shoots grow to 19-21cm, leave 2-7 strong shoots as main branches. When the main branches grow 6-7 pairs of lateral branches, spray the plant with the first aqueous solution of Uncaria rhynchophylla quality-inducing agent. (3) When the main branch grows to 15-17 pairs of lateral branches, pinch off the top and spray the plant with a second aqueous solution of Uncaria rhynchophylla quality-inducing agent. (4) Harvest the 1-6 pairs of lateral branches at the base in June-July for medicinal purposes, and harvest the remaining lateral branches in mid-to-late November; The phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer, by weight, comprises: 15-25 parts of Uncaria rhynchophylla stems and leaves (3:1 weight ratio), 6-12 parts of Luo Han Guo (monk fruit) extraction waste or residual fruit, 5-11 parts of tung oil bran, 5-11 parts of earthworm excrement, 5-11 parts of calcium magnesium phosphate fertilizer, 4-10 parts of silkworm residue, 4-10 parts of soybean meal, 3-9 parts of biochar, 2-4 parts of sugarcane filter mud, 3-9 parts of potassium humate, 0.1-0.3 parts of Pseudomonas fluorescens, and Bacillus mucilaginosus. 0.09-0.11 parts, 0.3-0.5 parts of phosphate-solubilizing bacteria, 0.1-0.3 parts of cellulose-solubilizing Vibrio butyric acid bacteria, 0.1-0.3 parts of Aspergillus niger, 0.1-0.3 parts of yeast, and 5-11 parts of slow-release compound fertilizer; wherein the effective viable count of the fluorescent Pseudomonas fluorescens is ≥200 million / ml, the effective viable count of the mucilage Bacillus is ≥5 billion / g, the effective viable count of the phosphate-solubilizing bacteria is ≥1.0×10 CFU / g, and the N:P:K ratio of the slow-release compound fertilizer is 15:8:18; Each kilogram of the above-mentioned Uncaria rhynchophylla extract inductive aqueous solution includes: 5-7 mg of strigolactone, 4-6 mg of folin, Benzylaminopurine 16-24mg, bio-organic vanadium 8-12mg, amino acid esters 10-15mg, amino acid zinc 16-24mg, amino acid chelated boron 81-99mg, organosilicon 89-111mg, amino acid chelated rare earth 800-1200mg, sugar alcohol calcium magnesium 180-220mg, potassium dihydrogen phosphate 800-1200mg.

2. The method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation according to claim 1, characterized in that, The phosphorus- and potassium-solubilizing compound slow-release microbial fertilizer, by weight, comprises: 20 parts of Uncaria rhynchophylla stems and leaves in a 3:1 weight ratio; 8.7 parts of Luo Han Guo extraction waste or residual fruit; 8 parts of tung oil bran; 8 parts of earthworm excrement; 8 parts of calcium magnesium phosphate fertilizer; 7 parts of silkworm residue; 7 parts of soybean meal; 6 parts of biochar; 3 parts of sucrose filter mud; 6 parts of potassium humate; 0.2 parts of Pseudomonas fluorescens; 0.1 parts of Bacillus mucilaginosus; 0.4 parts of phosphate-solubilizing bacteria; 0.2 parts of Vibrio butyricum fibrosum; 0.2 parts of Aspergillus niger; 0.2 parts of yeast; and 8 parts of slow-release compound fertilizer. The effective viable count of Pseudomonas fluorescens is ≥200 million / ml; the effective viable count of Bacillus mucilaginosus is ≥5 billion / g; the effective viable count of phosphate-solubilizing bacteria is ≥1.0×10 CFU / g; and the N:P:K ratio of the slow-release compound fertilizer is 15:8:

18.

3. The method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation according to claim 1 or 2, characterized in that, The preparation method of the phosphorus and potassium solubilizing compound slow-release microbial fertilizer is as follows: S1. Weigh out the following ingredients according to the specified ratio: Uncaria rhynchophylla stems and leaves, Luo Han Guo extract residue or fruit residue, tung oil bran, calcium magnesium phosphate fertilizer, silkworm residue, soybean meal, and sugarcane filter mud. Mix them together. The Uncaria rhynchophylla stems and leaves are crushed, and the Luo Han Guo extract residue or fruit residue is crushed. S2. Add Vibrio butyricum, Aspergillus niger, and yeast according to the formula, and add water to the moisture content to 39%-41%. Carry out rainproof composting and fermentation. During the fermentation process, when the temperature in the center reaches 50℃, turn the pile until it is fully decomposed. S3. Add biochar, earthworm excrement, potassium humate, Bacillus mucilaginosus, and phosphate-solubilizing bacteria according to the formula, maintain the moisture content at 24%-26%, and carry out composting culture. S4. Before use, add fluorescent Pseudomonas aeruginosa and slow-release compound fertilizer according to the ratio.

4. The method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation according to claim 3, characterized in that, Each kilogram of the above-mentioned Uncaria rhynchophylla extract inducing aqueous solution comprises: 6 mg of strigolactone, 5 mg of folinic acid, 20 mg of benzylaminopurine, 10 mg of bio-organic vanadium, 12 mg of amino acid ester, 20 mg of amino acid zinc, 90 mg of amino acid chelated boron, 100 mg of organosilicon, 1000 mg of amino acid chelated rare earth, 200 mg of sugar alcohol calcium magnesium, and 1000 mg of potassium dihydrogen phosphate.

5. The method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation according to claim 1 or 4, characterized in that, The preparation method of the aqueous solution of the Uncaria rhynchophylla extract inducer is as follows: S5. Weigh out strigolactone and benzylaminopurine according to the ratio, add acetone until completely dissolved, and then dilute with distilled water to form the first mother liquor. S6. Weigh out the following ingredients according to the specified ratio: leaf extract, organic vanadium, amino acid lipids, zinc amino acid chelated boron, organosilicon, rare earth amino acid chelated sugar alcohol, calcium magnesium, and potassium dihydrogen phosphate. Mix them and dissolve them in distilled water to form the second mother liquor. S7. Combine the first mother liquor and the second mother liquor, and dilute with distilled water to the required concentration to obtain an aqueous solution of Uncaria rhynchophylla extract inducing agent.

6. The method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation according to claim 1, characterized in that, The method for preparing the bio-organic vanadium is as follows: S8. Select plump soybeans, soak them in a 0.02% potassium permanganate solution for 1 minute, take them out, rinse them with distilled water to wash off the potassium permanganate on the surface of the soybeans, then soak them in 45℃ hot water for 20 minutes, take them out, drain the surface water, and set aside. S9. Soak soybeans in a sodium metavanadate aqueous solution with a concentration of 780mg / L-820mg / L, and keep them in a constant temperature biological incubator at 40℃ for 8 hours. Take them out, place them in a germination box, spread them out to a thickness of 2-3cm, cover them, and place them in a constant temperature biological incubator at 28℃ for 72 hours in the dark. Open the lid every day and sprinkle the same concentration of sodium metavanadate aqueous solution, and roll the soybeans to keep the soybean surface moist. S10. After the soybeans sprout, rinse the soybean sprouts with tap water, then rinse them with distilled water to wash away the sodium metavanadate on the surface of the soybean sprouts. Then dry them in a hot air drying oven at 60°C until constant weight, pulverize the soybeans, and obtain soybean tooth powder containing organic vanadium.

7. The method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation according to claim 1, characterized in that, During the plant's growth period, maintain a relative soil humidity of 60-80%.

8. The method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation according to claim 1, characterized in that, In step (3), 14-16 days after the second spraying of the Uncaria rhynchophylla quality-inducing agent on the plant, the plant is sprayed with the Uncaria rhynchophylla quality-inducing agent aqueous solution for the third time.

9. The method for increasing the yield of Uncaria rhynchophylla stems and promoting alkaloid accumulation according to claim 1, characterized in that, In step (2), leave 5-6 strong buds as main branches.

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