A cultivation method for increasing the puerarin content of Pueraria lobata

Abstract

The present invention relates to the technical field of kudzu cultivation, and in particular to a cultivation method for increasing the puerarin content of kudzu, the method comprising site selection for planting and fertilization management; wherein the fertilization management specifically comprises applying base fertilizer and topdressing, specifically comprising: (1) applying base fertilizer before planting kudzu seedlings and after land preparation; (2) applying topdressing with a special formula fertilizer for kudzu at the initial stage of kudzu tuber swelling, the special formula fertilizer for kudzu mainly comprising the following raw materials in parts by weight: 25-30 parts of urea, 15-20 parts of monoammonium phosphate, 30-50 parts of potassium sulfate, 0.3-0.5 parts of ammonium molybdate, 0.3-0.7 parts of high iron polypeptide fertilizer and 10-15 parts of auxiliary materials; through the method, kudzu with high puerarin content can be obtained, and the kudzu yield is high.

Description

[Technical field]

[0001] The invention relates to the technical field of kudzu vine cultivation, and in particular to a cultivation method for improving the puerarin content of kudzu vine. [Background technology]

[0002] Pueraria lobata is a plant of the genus Pueraria in the Leguminosae family. It is a commonly used Chinese medicinal material in my country and is one of the first plants recognized by the National Health Commission of the People's Republic of China as both a medicine and food. It is known as "Northern Ginseng and Southern Pueraria" and "Asian Ginseng" and has a broad market prospect. The "Pharmacopoeia of the People's Republic of China" (2020 edition) clearly stipulates that Pueraria lobata is the source of authentic medicinal products in my country. Puerarin is the main active substance of Pueraria lobata and is an important indicator for evaluating the quality of Pueraria lobata. Puerarin is mainly extracted and separated from plants of the genus Pueraria in the Leguminosae family. Its molecular formula is C 21 H 20 O9, whose chemical name is 7,4′-dihydroxy-8-β-D-glucosyl isoflavone, has the effects of dilating blood vessels, inhibiting platelet aggregation, and lowering blood pressure. It is mainly used clinically for cancer prevention, anti-oxidation, and treatment of cardiovascular and cerebrovascular diseases.

[0003] The Chinese Pharmacopoeia stipulates that the puerarin content of kudzu must not be less than 0.3%. As a major kudzu cultivation province, most of the kudzu varieties cultivated artificially in Guangxi are local varieties that have been planted for a long time and a small number of varieties selected and bred by scientific research institutions. The common characteristics of these kudzu varieties are enlarged tubers, rapid growth, and high starch content. However, tests have found that under the current kudzu cultivation model, the puerarin content of annual kudzu tubers is low, which does not meet the requirement of puerarin content ≥ 0.3% in the Chinese Pharmacopoeia Volume I, which greatly restricts the development of the kudzu industry.

[0004] In view of this, the present application proposes a cultivation method for increasing the puerarin content of Pueraria lobata to solve the above problems. [Summary of the invention]

[0005] In view of the above, it is necessary to provide a cultivation method for increasing the puerarin content of kudzu vine, by which kudzu vine with high puerarin content and high kudzu vine yield can be obtained.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] A cultivation method for increasing the puerarin content of kudzu vine, the method comprising selecting a site for seedling planting and fertilizing management;

[0008] The fertilization management includes applying base fertilizer and topdressing, specifically: (1) applying base fertilizer before planting kudzu seedlings and after land preparation (i.e., deep plowing of the land); (2) applying topdressing with a special formula fertilizer for kudzu in the early stage of kudzu tuber enlargement, wherein the special formula fertilizer for kudzu is mainly composed of the following raw materials in parts by weight: 25-30 parts of urea, 15-20 parts of monoammonium phosphate, 30-50 parts of potassium sulfate, 0.3-0.5 parts of ammonium molybdate, 0.3-0.7 parts of high-iron polypeptide fertilizer, and 10-15 parts of auxiliary materials.

[0009] Furthermore, the special formula fertilizer for kudzu vine is prepared by the following method: urea, monoammonium phosphate, potassium sulfate, ammonium molybdate, high-iron polypeptide fertilizer and auxiliary materials are crushed separately, mixed, and stirred evenly, and then granulated, dried, and sieved to obtain the special formula fertilizer for kudzu vine.

[0010] Furthermore, the high-iron polypeptide fertilizer is prepared by the following method: separating blood cells from animal blood, enzymatically hydrolyzing the collected blood cells, then selecting a high-iron polypeptide stock solution through molecular screening, and concentrating the high-iron polypeptide stock solution, spray-drying the concentrated solution obtained after concentration, and then obtaining the high-iron polypeptide fertilizer.

[0011] Furthermore, the enzymatic hydrolysis is specifically performed by using a protease.

[0012] Furthermore, the auxiliary material is one of humic acid and biochemical fulvic acid or a combination thereof; the organic matter content in the auxiliary material is greater than 70%.

[0013] Furthermore, the site selection for planting seedlings specifically includes: selecting fertile and loose humus soil or sandy loam with good drainage as the planting site, removing weeds and deep plowing the land; and then planting the kudzu seedlings in the treated plots.

[0014] Furthermore, the base fertilizer is mainly composed of the following raw materials in parts by weight: 15-20 parts of rapeseed cake, 3-8 parts of sodium selenite, 5-10 parts of wood ash, 2-5 parts of humic acid, 6-12 parts of peanut straw and 3-9 parts of bone meal.

[0015] Furthermore, the special formula fertilizer for Pueraria lobata is applied by root application, specifically: dig a pit with a diameter of 8-12cm and a depth of 12-15cm around the roots of the Pueraria lobata seedlings, fill the pit with soil so that the depth of the pit reaches 6-9cm, then put the special formula fertilizer for Pueraria lobata into the pit, then take soil and put it into the pit to fill the pit, cover the soil around the roots of the Pueraria lobata seedlings, and sprinkle appropriate amount of water to moisten the soil.

[0016] Furthermore, the application amount of the kudzu vine special fertilizer is 25-50 kg per acre of roots.

[0017] The present invention has at least the following beneficial effects:

[0018] 1. The cultivation method of the present application can effectively increase the content of puerarin and starch in Pueraria lobata, while ensuring the yield of Pueraria lobata, and can obtain high-quality Pueraria lobata with high puerarin and high starch content.

[0019] 2. The cultivation method of the present application mainly includes site selection for planting and fertilization management. First, a suitable planting site is selected, soil pretreatment is performed, and base fertilizer is applied. The base fertilizer of the present application also includes sodium selenite. During the cultivation process, the applicant found that the base fertilizer contains selenium, and its puerarin content is higher than that of the base fertilizer without selenium. This may be because the selenium-rich soil can enhance the transport and absorption capacity of the roots of Pueraria lobata for puerarin, so that the roots are enriched with puerarin. Therefore, the base fertilizer prepared by the applicant is not only rich in various fertilizers with nutrients required for the growth of Pueraria lobata, but also adds sodium selenite, in order to increase the content of puerarin.

[0020] 3. The cultivation method of the present application also includes fertilization management, which adopts a special formula fertilizer for kudzu vine, including N, P2O5 and K2O, wherein N is provided by urea (containing N ≥ 46%), N and P2O5 are provided by monoammonium phosphate (containing N ≥ 10%, containing P2O5 ≥ 55%), and K2O is provided by potassium sulfate (containing K2O ≥ 50%). In addition, it also includes a high-iron polypeptide fertilizer rich in polypeptides, which is a powdered fertilizer produced by advanced enzyme hydrolysis and other biotechnology, contains rich polypeptides and porphyrin iron. In addition, the special fertilizer of the present application also includes Mo element, which is conducive to biological nitrogen fixation of kudzu vine. The fertilizer of the present application has a reasonable formula and comprehensive nutrition, and can effectively promote the growth of kudzu vine tubers. In addition, the applicant found that the puerarin content and starch content of kudzu vine obtained by fertilizer cultivation of the present application were greatly improved, the quality of kudzu vine was good, and the market competitiveness of kudzu vine was improved.

[0021] 4. The applicant also found during the cultivation process that the method of fertilization has a great influence on the puerarin content of kudzu vine. In the present application, topdressing is performed by applying solid fertilizer to the roots. The timing of topdressing is the early stage of kudzu vine tuber expansion. At this time, the kudzu vine tubers can actively absorb nutrients brought from the roots, and puerarin is rapidly enriched. Previous studies have found that foliar fertilization has a better effect on the accumulation of certain effective ingredients of kudzu vine, such as selenium. This is because foliar fertilization can directly supply nutrients to plants, and the nutrient absorption and conversion is faster than the roots, which can meet the needs of plants in a timely manner. In the present application, experiments have found that foliar fertilization is used. The purpose of increasing the puerarin content cannot be achieved. The applicant speculates that it may be caused by the following two factors: first, after the raw material formula of the present application is prepared into a liquid fertilizer, porphyrin iron is insoluble in water, and the liquid fertilizer no longer contains porphyrin iron. In the process of foliar fertilization using the liquid fertilizer, the puerarin content of the fertilizer lacking porphyrin iron will be reduced, which is the content verified by previous experiments; second, kudzu root is drought-resistant but not water-resistant. The initial stage of kudzu root expansion is a period of rapid growth. The water demand at this stage is not very large. Foliar fertilization is generally prepared with liquid fertilizer, which is not suitable for kudzu at this growth stage, resulting in a decrease in puerarin content. [Specific implementation method]

[0022] In order to make the above-mentioned purpose, features and advantages of the present invention more obvious and easy to understand, the specific implementation of the present invention is described in detail below. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific implementation disclosed below.

[0023] Embodiment 1:

[0024] This embodiment provides a method for increasing the puerarin content of Pueraria lobata, the method comprising the following steps:

[0025] (1) Site selection for planting: Select fertile, loose, well-drained humus soil or sandy loam as the planting site, remove weeds, and deep-plow the land; then plant the kudzu seedlings in the treated plots;

[0026] (2) Fertilization management: including the application of base fertilizer and topdressing, specifically: 1) applying base fertilizer before planting kudzu seedlings and after land preparation; 2) applying topdressing with a special formula fertilizer for kudzu in the early stage of kudzu root expansion, the special formula fertilizer for kudzu is applied to the roots, 25 kg per mu of root application, specifically: digging a pit with a diameter of 8 cm and a depth of 12 cm around the roots of the kudzu seedlings, filling the pit with soil to make the pit depth reach 6 cm, then throwing the special formula fertilizer for kudzu into the pit, and then taking soil and throwing it into the pit to fill the pit, and covering the roots of the kudzu seedlings with soil, and applying an appropriate amount of water to moisten the soil; the base fertilizer mainly consists of the following raw materials in parts by weight: rapeseed cake 15 3 parts, sodium selenite 3 parts, plant ash 5 parts, humic acid 2 parts, peanut straw 6 parts and bone meal 3 parts; the kudzu vine special formula fertilizer is mainly composed of the following raw materials by weight: 25 parts of urea, 15 parts of monoammonium phosphate, 30 parts of potassium sulfate, 0.3 parts of ammonium molybdate, 0.3 parts of high iron polypeptide fertilizer and 10 parts of auxiliary materials; wherein the high iron polypeptide fertilizer is prepared by the following method: blood cells are separated from animal blood, the collected blood cells are enzymatically hydrolyzed by protease, and then the high iron polypeptide stock solution is selected by molecular screening, and the high iron polypeptide stock solution is concentrated, and the concentrated solution obtained after concentration is spray-dried to obtain the high iron polypeptide fertilizer; the auxiliary material is humic acid, and the organic matter content in the auxiliary material is greater than 70%;

[0027] The above-mentioned special formula fertilizer for kudzu vine is prepared by the following method: urea, monoammonium phosphate, potassium sulfate, ammonium molybdate, high-iron polypeptide fertilizer and auxiliary materials are crushed separately, mixed, and fully stirred, and then granulated, dried, and sieved to obtain the special formula fertilizer for kudzu vine.

[0028] Embodiment 2:

[0029] This embodiment provides a method for increasing the puerarin content of Pueraria lobata, the method comprising the following steps:

[0030] (1) Site selection for planting: Select fertile, loose, well-drained humus soil or sandy loam as the planting site, remove weeds, and deep-plow the land; then plant the kudzu seedlings in the treated plots;

[0031] (2) Fertilization management: including the application of base fertilizer and topdressing, specifically: 1) applying base fertilizer before planting kudzu seedlings and after land preparation; 2) applying topdressing with a special formula fertilizer for kudzu in the early stage of kudzu root expansion, the special formula fertilizer for kudzu is applied to the roots, 35 kg per mu of root application, specifically: digging a pit with a diameter of 10 cm and a depth of 13 cm around the roots of the kudzu seedlings, filling the pit with soil to make the pit depth reach 8 cm, then throwing the special formula fertilizer for kudzu into the pit, and then taking soil and throwing it into the pit to fill the pit, and covering the roots of the kudzu seedlings with soil, and applying an appropriate amount of water to moisten the soil; the base fertilizer mainly consists of the following raw materials in parts by weight: 17 parts of rapeseed cake, 6 parts of sodium selenite, and 10 parts of turmeric. 7 parts of wood ash, 4 parts of humic acid, 8 parts of peanut straw and 6 parts of bone meal; the special formula fertilizer for kudzu root mainly consists of the following raw materials in parts by weight: 27 parts of urea, 17 parts of monoammonium phosphate, 40 parts of potassium sulfate, 0.4 parts of ammonium molybdate, 0.5 parts of high-iron polypeptide fertilizer and 12 parts of auxiliary materials; wherein the high-iron polypeptide fertilizer is prepared by the following method: blood cells are separated from animal blood, the collected blood cells are enzymatically hydrolyzed by protease, and then the high-iron polypeptide stock solution is selected by molecular screening, and the high-iron polypeptide stock solution is concentrated, and the concentrated solution obtained after concentration is spray-dried to obtain the high-iron polypeptide fertilizer; the auxiliary materials are composed of humic acid and biochemical fulvic acid in a mass ratio of 1:1, and the organic matter content in the auxiliary materials is greater than 70%;

[0032] The above-mentioned special formula fertilizer for kudzu vine is prepared by the following method: urea, monoammonium phosphate, potassium sulfate, ammonium molybdate, high-iron polypeptide fertilizer and auxiliary materials are crushed separately, mixed, and fully stirred, and then granulated, dried, and sieved to obtain the special formula fertilizer for kudzu vine.

[0033] Embodiment 3:

[0034] This embodiment provides a method for increasing the puerarin content of Pueraria lobata, the method comprising the following steps:

[0035] (1) Selecting a site for planting seedlings: Select fertile, loose, well-drained humus soil or sandy loam as the planting site, remove weeds, deep-plow the land, and then apply base fertilizer; then plant the kudzu seedlings in the treated plots;

[0036] (2) Fertilization management: including the application of base fertilizer and topdressing, specifically: 1) applying base fertilizer before planting kudzu seedlings and after land preparation; 2) applying topdressing with a special formula fertilizer for kudzu in the early stage of kudzu root expansion, the special formula fertilizer for kudzu is applied to the roots, 50 kg per mu of root application, specifically: digging a pit with a diameter of 12 cm and a depth of 15 cm around the roots of the kudzu seedlings, filling the pit with soil to make the pit depth reach 9 cm, then putting the special formula fertilizer for kudzu into the pit, and then taking soil and putting it into the pit to fill the pit, and covering the roots of the kudzu seedlings with soil, and applying an appropriate amount of water to moisten the soil; the base fertilizer mainly consists of the following raw materials in parts by weight: 20 parts of rapeseed cake, 8 parts of sodium selenite, 10 parts of plant ash, 5 parts of humic acid, 12 parts of peanut straw and 9 parts of bone meal; the special formula fertilizer for kudzu root mainly consists of the following raw materials in parts by weight: 30 parts of urea, 20 parts of monoammonium phosphate, 50 parts of potassium sulfate, 0.5 parts of ammonium molybdate, 0.7 parts of high-iron polypeptide fertilizer and 15 parts of auxiliary materials; wherein the high-iron polypeptide fertilizer is prepared by the following method: separating blood cells from animal blood, enzymolyzing the collected blood cells with protease, then screening out high-iron polypeptide stock solution through molecular screening, concentrating the high-iron polypeptide stock solution, spray drying the concentrated solution obtained after concentration, and obtaining the high-iron polypeptide fertilizer; the auxiliary material is biochemical fulvic acid, and the organic matter content in the auxiliary material is greater than 70%;

[0037] The above-mentioned special formula fertilizer for kudzu vine is prepared by the following method: urea, monoammonium phosphate, potassium sulfate, ammonium molybdate, high-iron polypeptide fertilizer and auxiliary materials are crushed separately, mixed, and fully stirred, and then granulated, dried, and sieved to obtain the special formula fertilizer for kudzu vine.

[0038] In order to illustrate the effect of this application, the applicant conducted the following experiments:

[0039] The applicant compared the kudzu yield, puerarin content and starch content of the following groups, which were specifically divided into the following groups:

[0040] Group 1: the cultivation method described in Example 3;

[0041] Group 2: The fertilization management used conventional fertilizers, i.e., nitrogen, phosphorus and potassium fertilizers (derived from urea, monoammonium phosphate and potassium sulfate), and the other methods were the same as those in Example 3;

[0042] The third group: sodium selenite was removed from the base fertilizer, and the other methods were the same as those in Example 3;

[0043] Group 4: The topdressing time was at the late stage of kudzu root expansion, and the other methods were the same as those in Example 3;

[0044] Group 5: The preparation method of high-iron polypeptide fertilizer is as follows: blood cells are separated from animal blood, the collected blood cells are enzymatically hydrolyzed by protease, and then a high-iron polypeptide stock solution is obtained by molecular screening, and the high-iron polypeptide stock solution is concentrated, and the precipitated porphyrin iron is removed after concentration, and the obtained concentrated solution is spray-dried to obtain the high-iron polypeptide fertilizer; other methods are the same as those in Example 3;

[0045] Group 6: The special formula fertilizer for kudzu root is prepared by the following method: urea, monoammonium phosphate, potassium sulfate, ammonium molybdate, high iron polypeptide fertilizer and auxiliary materials are crushed respectively, mixed, 5-10 times the mass of water of the above raw materials are added, and stirred well, filtered to obtain the special formula fertilizer for kudzu root, and the fertilization method is foliar fertilization; other methods are the same as those in Example 3;

[0046] Group 7: The timing of topdressing was when the kudzu vines were in their vigorous growth period, and the other methods were the same as those in Example 3.

[0047] The cultivation effects of each group of Pueraria lobata are shown in Table 1:

[0048] Table 1 Comparison of cultivation effects of Pueraria lobata in each group

[0049]

[0050]

[0051] According to the results in Table 1, the kudzu vine obtained by the cultivation method of the present application not only has a high yield, but also has high puerarin content and starch content. The kudzu vine obtained by using the fertilizer and fertilization method described in the present application has good quality. The second group used conventional fertilizers for application, and the yield and quality were poor. The third group removed sodium selenite from the base fertilizer, resulting in a sharp drop in puerarin content. This may be because selenium can promote the enrichment of puerarin content in the transport. The fourth and seventh groups changed the timing of topdressing, and the effects described in the present application could not be achieved. This shows that the timing of fertilization also has a great influence on the growth of kudzu vine. How to choose the appropriate time for topdressing is crucial to the growth of kudzu vine. The high-iron polypeptide fertilizer of the fifth group removed porphyrin iron, and the sixth group was formulated into liquid fertilizer, resulting in a decrease in puerarin content in the fertilizer lacking porphyrin iron.

[0052] The above examples only express several embodiments of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the present invention. It should be pointed out that, for a person of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the attached claims.

Claims

1. A cultivation method for increasing the puerarin content of Pueraria lobata, characterized in that: The method includes site selection, planting and fertilizer management; The fertilization management includes applying base fertilizer and topdressing, specifically: (1) applying base fertilizer before planting kudzu seedlings and after land preparation; (2) applying topdressing with a special formula fertilizer for kudzu in the early stage of kudzu root expansion, wherein the special formula fertilizer for kudzu is composed of the following raw materials in parts by weight: 25-30 parts of urea, 15-20 parts of monoammonium phosphate, 30-50 parts of potassium sulfate, 0.3-0.5 parts of ammonium molybdate, 0.3-0.7 parts of high iron polypeptide fertilizer and 10-15 parts of auxiliary materials; The high-iron polypeptide fertilizer is prepared by the following method: separating blood cells from animal blood, enzymatically hydrolyzing the collected blood cells, then selecting a high-iron polypeptide stock solution through molecular screening, concentrating the high-iron polypeptide stock solution, and spray-drying the concentrated solution obtained after concentration to obtain the high-iron polypeptide fertilizer; the auxiliary material is one of humic acid and biochemical fulvic acid or a combination thereof; the base fertilizer is composed of the following raw materials in parts by weight: 15-20 parts of rapeseed cake, 3-8 parts of sodium selenite, 5-10 parts of wood ash, 2-5 parts of humic acid, 6-12 parts of peanut straw and 3-9 parts of bone meal; the special formula fertilizer for kudzu is applied by root application.

2. The method according to claim 1, characterized in that The special formula fertilizer for kudzu vine is prepared by the following method: urea, monoammonium phosphate, potassium sulfate, ammonium molybdate, high-iron polypeptide fertilizer and auxiliary materials are crushed, mixed, and fully stirred, and then granulated, dried, and sieved to obtain the special formula fertilizer for kudzu vine.

3. The method according to claim 1, characterized in that: The enzymatic hydrolysis specifically comprises enzymatic hydrolysis treatment with protease.

4. The method according to claim 1, characterized in that The site selection and planting specifically includes: selecting fertile and loose humus soil or sandy loam with good drainage as the planting site, removing weeds and deep plowing the land; and then planting the kudzu seedlings in the treated plots.

5. The method according to claim 1, characterized in that The special formula fertilizer for Pueraria lobata is applied through the roots, specifically: dig a pit with a diameter of 8-12cm and a depth of 12-15cm between two Pueraria lobata plants, fill the pit with soil so that the depth of the pit reaches 6-9cm, then put the special formula fertilizer for Pueraria lobata into the pit, then take soil and put it into the pit to fill it up, cover the soil around the roots of the Pueraria lobata seedlings, and apply appropriate amount of water to moisten the soil.

6. The method according to claim 1, characterized in that The application amount of the kudzu vine special fertilizer is 25-50kg per mu of root.

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