A method for differentiating formula fertilization of scutellaria baicalensis

Abstract

This invention discloses a differentiated fertilization method for Scutellaria baicalensis, belonging to the field of traditional Chinese medicinal herb cultivation technology. This method selects a corresponding ratio from multiple predetermined nitrogen, phosphorus, and potassium fertilizer application ratios based on the accumulation requirements of the target medicinal components, thereby achieving targeted regulation of different medicinal components. The multiple predetermined nitrogen-phosphorus-potassium ratios include a first ratio that increases baicalin, a second ratio that increases baicalein, a third ratio that increases wogonin, and a fourth ratio that increases wogonein. This invention allows for flexible selection of ratios according to production targets, achieving precise regulation of different medicinal components while simultaneously increasing Scutellaria baicalensis yield, and has good prospects for widespread application.

Description

Technical Field

[0001] This invention relates to the field of Chinese medicinal herb cultivation technology, specifically to a differentiated fertilizer application method for Scutellaria baicalensis. Background Technology

[0002] Scutellaria baicalensis Georgi is a perennial herb belonging to the genus Scutellaria in the family Lamiaceae. Its dried root is used medicinally and has the effects of clearing heat and drying dampness, purging fire and detoxifying, stopping bleeding and calming the fetus. It is one of the commonly used bulk Chinese medicinal materials in my country.

[0003] The main medicinal components of Scutellaria baicalensis include flavonoids such as baicalin, baicalein, wogonin, and wogonin. Among them, baicalin is listed as a quality control indicator component of Scutellaria baicalensis in the Chinese Pharmacopoeia. Scientific fertilization is a key aspect of the ecological cultivation of medicinal herbs. Studies have shown that using nitrogen, phosphorus, and potassium (NPK) formulated fertilizers instead of single-element fertilizers is beneficial for promoting the growth of medicinal plants and the accumulation of effective components, improving the quality and yield of medicinal herbs, while maintaining the balance of the soil micro-ecosystem.

[0004] However, existing technologies have the following shortcomings: There is a lack of research on fertilization for Scutellaria baicalensis, especially a lack of systematic research on Scutellaria baicalensis fertilization. Standardized cultivation lacks scientific fertilization data to support it. Current Scutellaria baicalensis cultivation generally suffers from blind fertilization and single-ratio fertilization, leading to unstable yields and large fluctuations in the content of effective components. Furthermore, traditional fertilization methods often aim for "high yield" or "high quality" with a fixed nitrogen, phosphorus, and potassium ratio, but research has found that different medicinal components accumulate in Scutellaria baicalensis according to different patterns. The differences in the medicinal materials mean that the demand for nitrogen, phosphorus, and potassium also vary. Fixed-ratio fertilization methods cannot simultaneously meet the accumulation needs of multiple medicinal components, making it difficult to achieve precise control over the quality of medicinal materials. In addition, existing technologies lack the concept of "fertilization on demand," that is, differentiated fertilization strategies that select different fertilizer ratios based on different production goals (such as focusing on increasing baicalin or baicalein). This makes it difficult to directionally control the content of specific medicinal components in Scutellaria baicalensis cultivation, and thus cannot meet the diversified demands of the Chinese medicinal materials market for medicinal materials of different quality and specifications.

[0005] Therefore, developing a differentiated fertilization method that can selectively use different nitrogen, phosphorus, and potassium ratios based on the accumulation requirements of the target medicinal components is of great significance for improving the yield and quality of Scutellaria baicalensis and promoting its standardized cultivation. Summary of the Invention

[0006] To address the aforementioned shortcomings in existing technologies, this invention provides a differentiated fertilization method for Scutellaria baicalensis. This method aims to selectively apply different ratios of nitrogen, phosphorus, and potassium fertilizers based on the accumulation requirements of the target medicinal components, thereby achieving targeted regulation of different medicinal components such as baicalin, baicalein, wogonin, and wogonin, while simultaneously increasing the yield of Scutellaria baicalensis.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A differentiated fertilizer application method for Scutellaria baicalensis involves selecting a corresponding ratio from multiple predetermined nitrogen, phosphorus, and potassium fertilizer application ratios based on the accumulation requirements of the target medicinal components, in order to achieve targeted regulation of different medicinal components. The multiple predetermined nitrogen, phosphorus, and potassium ratios include a first ratio that increases baicalin, a second ratio that increases baicalein, a third ratio that increases wogonin, and a fourth ratio that increases wogonein.

[0008] Furthermore, the nitrogen fertilizer is urea, the phosphate fertilizer is superphosphate, and the potassium fertilizer is potassium sulfate.

[0009] Furthermore, fertilization includes two applications: basal fertilizer and topdressing. The basal fertilizer consists of all the phosphorus fertilizer, all the potassium fertilizer, and 50% of the nitrogen fertilizer. The topdressing is applied the following year during the greening and growth period of Scutellaria baicalensis, with the remaining 50% of the nitrogen fertilizer applied.

[0010] Furthermore, the application rate of nitrogen fertilizer is 106.3–318.7 kg / hm². 2 The application rate of phosphate fertilizer is 126.5–382.2 kg / hm². 2 The application rate of potassium fertilizer is 88–264 kg / hm². 2 .

[0011] Furthermore, the first ratio is: nitrogen fertilizer application rate is 215.5 kg / hm². 2 The application rate of phosphate fertilizer was 254.8 kg / hm², and the application rate of potassium fertilizer was 176 kg / hm².

[0012] Furthermore, the second ratio is: nitrogen fertilizer application rate of 106.3 kg / hm². 2 The application rate of phosphate fertilizer was 126.5 kg / hm². 2 The application rate of potassium fertilizer is 176 kg / hm. 2 .

[0013] Furthermore, the third ratio is: nitrogen fertilizer application rate of 215.5 kg / hm². 2 The application rate of phosphate fertilizer was 126.5 kg / hm². 2 The application rate of potassium fertilizer is 88 kg / hm. 2 .

[0014] Furthermore, the fourth ratio is: nitrogen fertilizer application rate of 215.5 kg / hm². 2 The application rate of phosphate fertilizer was 254.8 kg / hm². 2 The application rate of potassium fertilizer is 88 kg / hm. 2 .

[0015] Furthermore, Scutellaria baicalensis is a biennial cultivated Scutellaria baicalensis.

[0016] Furthermore, the base fertilizer should be applied before or on the day of sowing Scutellaria baicalensis, and the topdressing fertilizer should be applied in early May of the following year during the greening and growth period of Scutellaria baicalensis.

[0017] Compared with the prior art, the present invention has the following beneficial effects: 1. This invention proposes a differentiated fertilization strategy that selectively employs different nitrogen, phosphorus, and potassium ratios based on the accumulation requirements of the target medicinal components. Through systematic field trials, the optimal fertilization ratio for enhancing different medicinal components was identified: for enhancing baicalin, the first ratio (i.e., N2P2K2 ratio: nitrogen fertilizer 215.5 kg / hm²) was used. 2 Phosphate fertilizer 254.8 kg / hm 2 Potassium fertilizer 176 kg / hm 2 To enhance baicalin production, a second ratio (i.e., N1P1K2 ratio: nitrogen fertilizer 106.3 kg / hm²) was used. 2 Phosphate fertilizer 126.5 kg / hm 2 Potassium fertilizer 176 kg / hm 2 To increase baicalin levels, a third ratio (i.e., N2P1K1 ratio: nitrogen fertilizer 215.5 kg / hm²) is used. 2 Phosphate fertilizer 126.5 kg / hm 2 Potassium fertilizer 88 kg / hm 2 To enhance baicalin production, a fourth ratio (i.e., N2P2K1 ratio: nitrogen fertilizer 215.5 kg / hm²) was adopted. 2 Phosphate fertilizer 254.8 kg / hm 2 Potassium fertilizer 88 kg / hm 2 This differentiated fertilization method can be flexibly selected according to production goals, achieving precise control of different medicinal components and meeting the diversified needs of the Chinese medicinal materials market for medicinal materials of different quality and specifications.

[0018] 2. The fertilization method described in this invention is simple to operate, has a wide range of fertilizer sources, and allows for clear timing of fertilization. It is easy to promote and apply in the main production areas of Scutellaria baicalensis and has good prospects for industrialization. Attached Figure Description

[0019] Figure 1 This is the HPLC chromatogram of the mixed reference standard and the Scutellaria baicalensis sample.

[0020] Wherein: A is the chromatogram of the mixed reference standard (containing baicalin, baicalein, wogonin, and wogonin); B is the chromatogram of the sample treated with Scutellaria baicalensis N3P2K2.

[0021] In the figure, 1 represents baicalin, 2 represents baicalein, 3 represents wogonin, and 4 represents wogonein. Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0023] In the description of this invention, it should be noted that the terms "first", "second", "third" and "fourth" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0024] Example 1, as a preferred embodiment of the present invention, the fertilization method of this embodiment is as follows.

[0025] This embodiment uses two-year-old Scutellaria baicalensis Georgi. The sample was collected from the demonstration base of the Chengde Comprehensive Experimental Station of the National Traditional Chinese Medicine Industry Technology System. It was identified as a perennial herb belonging to the Scutellaria genus of the Lamiaceae family.

[0026] The experimental site for this embodiment was located at the Chengde Comprehensive Experimental Station of the National Traditional Chinese Medicine Industry Technology System (geographical coordinates: 41°25′N, 117°34′E). The area has distinctive topographic features, with an average altitude of 607m and a temperate continental monsoon climate, with an average annual temperature of 6.7℃ and annual precipitation of 510.7mm. Soil fertility testing revealed the following basic soil fertility indicators at the experimental site: organic matter content 7.11g / kg, available nitrogen content 72.7mg / kg, available phosphorus content 7.0mg / kg, and available potassium content 91.6mg / kg.

[0027] This embodiment employs a "3414" fertilization scheme to conduct a field trial of Scutellaria baicalensis, which involves three factors (nitrogen (N), phosphorus (P2O5), and potassium (K2O)) at four levels, with 14 fertilization treatments, each with three replicates, for a total of 42 experimental plots, randomly distributed, each plot being 50 m². 2 The fertilization levels are shown in Table 1. The experimental fertilizers were: urea (containing 46% N) for nitrogen, superphosphate (containing 46% N) and superphosphate (containing 18% P2O5) for phosphate, and potassium sulfate (containing 2% K2O5) for potassium. Scutellaria baicalensis seeds were sown on May 9, 2023. The fertilization plan followed the nutrient management principle of "one-time basal fertilizer as the main component, supplemented by periodic topdressing." Specifically, phosphate, potassium, and 50% of the nitrogen fertilizer were applied as basal fertilizer in one application; the remaining 50% of the nitrogen fertilizer was applied as topdressing on May 9, 2024.

[0028] Table 1. Treatment and fertilization amount in the “3414” experiment.

[0029]

[0030] Sampling for this experiment was conducted on September 20, 2024, collecting plant and soil samples from each experimental plot. An "S-shaped sampling method" was implemented within each plot, with each sampling point covering an area of ​​1 square meter. 2 (1m×1m), with 3 replicate sampling points set up for each treatment (i.e., 3 different locations in each plot). Complete plant samples were collected from each plot, and topsoil samples were also collected at a depth of 20cm as samples.

[0031] This embodiment measures the growth indicators and yield of Scutellaria baicalensis from collected samples. The specific methods are as follows: 1. Determination of growth indicators of Scutellaria baicalensis: The diameter of the taproot of Scutellaria baicalensis was measured, and the fresh weight of the roots of a single plant was weighed using an electronic balance. The total root length and taproot diameter of Scutellaria baicalensis were determined using a root scanner.

[0032] 2. Scutellaria baicalensis yield determination: The rhizome tissues of the collected Scutellaria baicalensis samples were brought back to the room, the soil attached to the rhizomes was removed, the rhizomes were rinsed with clean water, the surface moisture of the rhizomes was dried, and then dried in an oven at 60℃-65℃ until constant weight. After drying, the rhizomes of each Scutellaria baicalensis plant were weighed using an electronic analytical balance, and the dry weight of the rhizomes of each plant was recorded. The yield of medicinal material per unit area was calculated by summing the dry weights of all individual plants in each replicate plot.

[0033] The results of the growth indicators and yield measurements of Scutellaria baicalensis are shown in Table 2.

[0034] Table 2. Growth indicators and yield of Scutellaria baicalensis under different nitrogen, phosphorus and potassium application levels.

[0035]

[0036]

[0037] Table 2 shows that different combinations of nitrogen, phosphorus, and potassium application increased the root length, root diameter, and fresh root weight of Scutellaria baicalensis to varying degrees. The N3P2K2 treatment showed significantly higher root fresh weight and yield than other treatment groups, with the largest increases compared to the control group (N0P0K0), at 32.01% and 32.96%, respectively. Considering both growth indicators and yield, the N2P2K2 and N3P2K2 treatments performed better.

[0038] In this embodiment, the contents of baicalin, baicalein, wogonin, and baicalein in each treated Scutellaria baicalensis sample were determined by HPLC. The specific method is as follows: 1. Reference standards and reagents: Baicalin (C 21 H 18 O 11Batch number: 21121404, purity ≥98%), flavonoids (C 15 H 10 O5, batch number: PS000472, purity ≥98%), baicalin (C 22 H 20 O 11 Batch number: PS011071, purity ≥98%), baicalin (C 16 H 12 O5 (batch number: PS011541, purity ≥98%) was purchased from Chengdu Pusi Biotechnology Co., Ltd. Methanol was chromatographically pure, water was ultrapure water, and phosphoric acid was analytically pure.

[0039] 2. Preparation of mixed reference solution: Accurately weigh appropriate amounts of baicalin, baicalein, wogonin, and wogonin reference standards, place them in the same volumetric flask, dissolve and dilute to the mark with methanol, shake well, and prepare a mixed reference solution containing 1.20 mg / mL baicalin, 0.28 mg / mL baicalein, 0.14 mg / mL wogonin, and 0.05 mg / mL wogonin.

[0040] 3. Preparation of test solution: Accurately weigh 0.10 g of dried rhizome sample powder (passed through No. 3 sieve) obtained from each fertilization level, place it in a microwave extraction vessel, add 3 mL of 70% ethanol solution, microwave extract for 45 min, cool to room temperature, filter, and collect the filtrate to obtain the test solution.

[0041] 4. Chromatographic conditions: The column was an Agilent ZORBAX SB-C18 (4.5×250mm, 5μm); the stationary phase was octadecylsilane-bonded silica gel; the mobile phase was methanol (A)-water-phosphoric acid (B) (47:53:0.2); the gradient elution program was: 0~18min, 48%~65%A; 18~25min, 65%A; 25~26min, 65~100%A; 26~28min, 100%A; column temperature 25℃; injection volume 10μL; flow rate 1 mL / min; detection wavelength 280nm; detection time 28min.

[0042] 5. System suitability test: Accurately pipette 10 μL of the mixed reference solution, inject and determine the chromatogram under the above chromatographic conditions, and record the results as follows. Figure 1 As shown. Among them. Figure 1 -A is the chromatogram of the mixed reference standards. Figure 1 -B is the chromatogram of the sample treated with Scutellaria baicalensis N3P2K2. Figure 1As can be seen, all analytes were well separated within 28 min, and the chromatographic peaks were symmetrical. Peak 1 in the figure is baicalin (retention time approximately 9.4 min), peak 2 is baicalein (retention time approximately 13.4 min), peak 3 is wogonin (retention time approximately 17.6 min), and peak 4 is wogonein (retention time approximately 23.2 min). The resolution between each chromatographic peak and its adjacent peak was greater than 1.5, and the theoretical plate number, calculated based on the baicalin peak, was not less than 5000, meeting the suitability requirements for the chromatographic system as stipulated in the Chinese Pharmacopoeia.

[0043] 6. Preparation of Standard Curve: Accurately pipette the mixed reference solution and dilute it stepwise with methanol to obtain a series of mixed reference solutions of different concentrations. Inject and determine the peak areas separately under the chromatographic conditions described above. Plot the standard curve with the concentration of each reference standard as the abscissa (X) and the peak area as the ordinate (Y) to obtain the regression equation. The results of the linear relationship investigation of each component are shown in Table 3. The results show that each compound has a good linear relationship within the set concentration range (R0). 2 All reached 0.9999).

[0044] Table 3. Results of linear relationship study of four medicinal components in Scutellaria baicalensis root.

[0045]

[0046] 7. Sample Content Determination: Take the Scutellaria baicalensis powder samples obtained at each fertilization level, prepare the test solution according to the method under "Preparation of Test Solution", inject and determine the sample under the above chromatographic conditions, record the peak area, and substitute it into the standard curve to calculate the content of each component. The determination results are shown in Table 4.

[0047] Table 4. Content of main effective components in Scutellaria baicalensis under different nitrogen, phosphorus and potassium application levels.

[0048]

[0049]

[0050] Table 4 shows that there are significant differences in the optimal fertilization ratios of different active ingredients: baicalin content was highest in N2P2K2 at 163.95 mg / g, an increase of 23.82% compared to the control group; baicalein content was highest in N1P1K2 at 41.04 mg / g, an increase of 29.52% compared to the control group; wogonin content was highest in N2P1K1 at 14.32 mg / g, an increase of 71.70% compared to the control group; and wogonein content was highest in N2P2K1 at 6.19 mg / g, an increase of 102.95% compared to the control group.

[0051] This result indicates that different active ingredients have different requirements for nitrogen, phosphorus, and potassium, and that a differentiated fertilization strategy can achieve targeted regulation of different ingredients.

[0052] Based on the above results, the overall effects of each preferred ratio in the differentiated fertilization method described in this invention are as follows: First ratio (N2P2K2: Nitrogen 215.5 kg / hm) 2 Phosphorus 254.8 kg / hm 2 Potassium 176 kg / hm 2 The highest content of baicalin was found in this variety, reaching 163.95 mg / g; the yield was also relatively high, reaching 832.99 kg / hm². 2 The root system is well developed, with a taproot diameter of 10.78 mm and a fresh root weight of 22.18 g.

[0053] The second ratio (N1P1K2: Nitrogen 106.3 kg / hm) 2 Phosphorus 126.5 kg / hm 2 Potassium 176 kg / hm 2 The highest content of baicalein was found in 41.04 mg / g; the content of wogonin was also relatively high, reaching 5.89 mg / g.

[0054] The third ratio (N2P1K1: Nitrogen 215.5 kg / hm) 2 Phosphorus 126.5 kg / hm 2 Potassium 88 kg / hm 2 The highest content of baicalin was found in 14.32 mg / g.

[0055] Fourth ratio (N2P2K1: Nitrogen 215.5 kg / hm) 2 Phosphorus 254.8 kg / hm 2 Potassium 88 kg / hm 2 The highest content of baicalein was 6.19 mg / g; the content of baicalin was also relatively high, reaching 14.12 mg / g.

[0056] Therefore, in production, the corresponding ratio can be selected from the above four preferred ratios according to the accumulation requirements of the target active ingredient, so as to achieve targeted regulation of different active ingredients.

[0057] Finally, it should be noted that the above embodiments are merely preferred embodiments of the present invention used to illustrate the technical solutions of the present invention, and are not intended to limit the invention, nor are they intended to limit the patent scope of the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. These modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention. That is to say, any changes or refinements made to the main design concept and spirit of the present invention that are not of substantial significance, but whose technical problems are still consistent with the present invention, should be included within the protection scope of the present invention. In addition, the direct or indirect application of the technical solutions of the present invention to other related technical fields are similarly included within the patent protection scope of the present invention.

Claims

1. A differentiated fertilizer application method for Scutellaria baicalensis, characterized in that, Based on the accumulation requirements of the target active ingredients, the corresponding ratio is selected from multiple predetermined nitrogen, phosphorus, and potassium fertilizer application ratios for application, in order to achieve targeted regulation of different active ingredients; the multiple predetermined nitrogen, phosphorus, and potassium ratios include a first ratio to increase baicalin, a second ratio to increase baicalein, a third ratio to increase wogonin, and a fourth ratio to increase wogonein.

2. The fertilization method according to claim 1, characterized in that, Nitrogen fertilizer is urea, phosphorus fertilizer is superphosphate, and potassium fertilizer is potassium sulfate.

3. The fertilization method according to claim 1, characterized in that, Fertilization includes two applications: basal fertilizer and topdressing. The basal fertilizer consists of all the phosphorus fertilizer, all the potassium fertilizer, and 50% of the nitrogen fertilizer. The topdressing is applied the following year during the greening and growth period of Scutellaria baicalensis, with the remaining 50% of the nitrogen fertilizer applied.

4. The fertilization method according to claim 1, characterized in that, The application rate of nitrogen fertilizer is 106.3–318.7 kg / hm². 2 The application rate of phosphate fertilizer is 126.5–382.2 kg / hm². 2 The application rate of potassium fertilizer is 88–264 kg / hm². 2 .

5. The fertilization method according to claim 1, characterized in that, The first ratio is: nitrogen fertilizer application rate of 215.5 kg / hm². 2 The application rate of phosphate fertilizer was 254.8 kg / hm. 2 Potassium fertilizer application rate: 176 kg / hm 2 .

6. The fertilization method according to claim 1, characterized in that, The second ratio is: nitrogen fertilizer application rate of 106.3 kg / hm². 2 The application rate of phosphate fertilizer was 126.5 kg / hm². 2 The application rate of potassium fertilizer is 176 kg / hm. 2 .

7. The fertilization method according to claim 1, characterized in that, The third ratio is: nitrogen fertilizer application rate of 215.5 kg / hm². 2 The application rate of phosphate fertilizer was 126.5 kg / hm². 2 The application rate of potassium fertilizer is 88 kg / hm. 2 .

8. The fertilization method according to claim 1, characterized in that, The fourth ratio is: nitrogen fertilizer application rate of 215.5 kg / hm². 2 The application rate of phosphate fertilizer was 254.8 kg / hm². 2 The application rate of potassium fertilizer is 88 kg / hm. 2 .

9. The fertilization method according to claim 1, characterized in that, Scutellaria baicalensis is a biennial cultivated Scutellaria baicalensis.

10. The fertilization method according to claim 3, characterized in that, The base fertilizer should be applied before or on the day of sowing Scutellaria baicalensis, and the top dressing should be applied in early May of the following year during the greening and growth period of Scutellaria baicalensis.

Images