A planting method for increasing the chromone content in Saposhnikovia divaricata

Abstract

The present invention provides a planting method for increasing the chromone content in Saposhnikovia divaricata, which combines a medium and trace element composition and a microbial preparation in the planting of Saposhnikovia divaricata. The composition is CaCl2, MgSO4, FeCl3, MnCl2, CuCl2, ZnCl2. The strain in the microbial preparation is Bacillus subtilis. The medium and trace element composition has a significant effect on the contents of prim-O-glucosylcimifugin, prim-O-glucosylcimifugin glycoside and 5-O-methylvisamminol glycoside in Saposhnikovia divaricata.

Description

Technical Field

[0001] The present invention relates to the technical field of microbial fertilizers, and particularly relates to the application of a microbial preparation and medium and trace elements in the planting of Saposhnikovia divaricata. Background Art

[0002] The root of Saposhnikovia divaricata (Turcz.) Schischk. of the Umbelliferae family is used as the dried root of Saposhnikovia divaricata. The roots of plants that have not bolted are dug in spring and autumn, the fibrous roots and sediment are removed, and then dried in the sun. Saposhnikovia divaricata has the effects of expelling wind and relieving exterior syndrome, eliminating dampness and alleviating pain, and stopping convulsions. It is used for cold headache, rheumatic arthralgia, rubella itching, and tetanus.

[0003] Saposhnikovia divaricata contains volatile oils, coumarins, chromones and other components, among which chromone compounds are the main components for Saposhnikovia divaricata to exert its medicinal effects. Prim-O-glucosylcimifugin and 5-O-methylvisammioside are often used as the index components in the Chinese Pharmacopoeia to evaluate the quality of Saposhnikovia divaricata, and the total amount of the two is required to be not less than 0.24%. The latest research found that 3 ’ The changes of the two components of 3-O-angeloyl-hamaudol and prim-O-glucosylcimifugin contribute greatly to distinguishing Saposhnikovia divaricata with different growth modes and years, and they can be considered as quality markers for controlling the growth process of Saposhnikovia divaricata. 3 ’ 3-O-angeloyl-hamaudol can be used as a substrate for peroxidase, and significantly improve the antioxidant capacity and eliminate excessive hydrogen peroxide through the action of the enzyme.

[0004] Wang Yingfan et al. (Journal of Jilin Agricultural University, 2006, 28(3): 289-291) compared and analyzed the active ingredients of cultivated Saposhnikovia divaricata of one and two years with the contents of 4 chromones and polysaccharides as indicators. The results showed that the sum of the contents of prim-O-glucosylcimifugin and 5-O-methylvisammioside in Saposhnikovia divaricata of one and two years were 0.481% and 0.506% respectively, both far exceeding the specified limits in the Chinese Pharmacopoeia. However, the contents of prim-O-glucosylcimifugin, prim-O-glucosylcimifugin and sec-O-glucosylhamaudol in Saposhnikovia divaricata of two years were significantly higher than those of one-year-old Saposhnikovia divaricata, and only the content of 5-O-methylvisammioside in one-year-old Saposhnikovia divaricata was significantly higher than that of two-year-old Saposhnikovia divaricata.

[0005] Extending the planting years of Saposhnikovia divaricata can increase the contents of most chromones, but it also increases the planting time cost and may lead to a decrease in the content of 5-O-methylvisammioside. Due to the limited cultivated land area in the authentic production area, in order to pursue the economic benefits of planting Saposhnikovia divaricata, it is necessary not to extend the planting years as much as possible, which requires research on how to increase the content of chromones in one-year-old Saposhnikovia divaricata.

[0006] Guo Xu et al. (Journal of South China Agricultural University, 2020, 41(4): 30-37) studied the differences in the quality of radix fangfengi cultivated from different origins and the relationship between soil factors and the quality of the medicinal materials. The results showed that the total amount of radix fangfengi chromones was significantly negatively correlated with soil electrical conductivity, available phosphorus content and available calcium content, and significantly positively correlated with available manganese content. Available phosphorus, available manganese and total phosphorus content can explain 71.8% of the total chromone information of radix fangfengi.

[0007] Plant growth requires balanced nutrition. In the growth, development and metabolism of medicinal plants, in addition to the role played by macroelements such as nitrogen, phosphorus and potassium, trace elements such as calcium, magnesium, iron, manganese and zinc are also indispensable elements. Although plants do not require large amounts of trace elements, they are crucial to plant yield and quality. Lack or excess of these elements will affect the root nutrition and physiological functions of plants, thereby affecting the accumulation of their effective chemical components, leading to reduced yield or quality of medicinal plants. Summary of the invention

[0008] The invention provides a planting method for increasing the content of chromones in siler. A medium and trace element composition and a microbial preparation are jointly applied to siler planting. The strain in the microbial preparation is Bacillus subtilis. The chromones capable of increasing the content are cimicifuga glycoside, cimicifuga glycoside, 5-O-methylvisaminol glycoside, chelidonol glycoside and 3'-O-angeloylchelidonol.

[0009] The composition of trace elements is CaCl2, MgSO4, FeCl3, MnCl2, CuCl2 and ZnCl2.

[0010] In the medium and trace element composition, CaCl is 300-1200 weight parts, MgSO is 60-180 weight parts, FeCl is 15-120 weight parts, MnCl is 10-80 weight parts, CuCl is 10-80 weight parts, and ZnCl is 5-40 weight parts. Preferably, CaCl is 300-900 weight parts, MgSO is 60-120 weight parts, FeCl is 15-60 weight parts, MnCl is 10-40 weight parts, CuCl is 10-40 weight parts, and ZnCl is 5-20 weight parts. More preferably, CaCl is 600 weight parts, MgSO is 90 weight parts, FeCl is 30 weight parts, MnCl is 20 weight parts, CuCl is 20 weight parts, and ZnCl is 10 weight parts.

[0011] The combination of trace elements has a significant effect on the contents of cimicifuga, cimicifuga glycosides and 5-O-methylvisaminol glycosides in Saposhnikovia divaricata.

[0012] The present invention unexpectedly discovers that, compared with the single use of medium and trace elements, the combined application of a medium and trace element composition and a microbial preparation can significantly increase the chromone content in Saposhnikovia divaricata.

[0013] The strain in the microbial preparation is selected from Bacillus Subtilis tkm-1, which is preserved in the General Microbiology Center of the China Committee for Culture Collection of Microorganisms, with the preservation number CGMCC No. 14950, and has been published in Patent CN108865949A.

[0014] Inoculate Bacillus Subtilis tkm-1 into LB medium and culture it at 30 - 32 °C for 24 - 48 h to obtain the Bacillus Subtilis tkm-1 culture solution.

[0015] Among them, the LB medium consists of 5 g / L of yeast powder, 10 g / L of peptone, 10 g / L of sodium chloride, 20 g / L of glucose, and 20 g / L of agar powder.

[0016] The microbial preparation also includes shrimp and crab shells, collagen, sodium chloride, compound amino acids, carbohydrates, compound fertilizers. The weight ratio of the strain culture solution to shrimp and crab shells, collagen, sodium chloride, compound amino acids, carbohydrates, compound fertilizers is 1:(1 - 10):(1 - 10):(0.1 - 1):(10 - 50):(1 - 20):(1 - 10), preferably 1:4:3:0.3:20:10:5.

[0017] The shrimp and crab shells are from the shells of any chitin-rich aquatic animals in rivers, lakes, and seas, and their dried products are fine particle powders passed through a 20 - 100 mesh sieve. Preferably, they are shrimp shell powder, crab shell powder, or shrimp and crab powder.

[0018] The collagen is selected from one or more of porcine skin collagen, fish skin collagen, and bovine skin collagen.

[0019] The main components of the compound amino acids are glycine, alanine, leucine, isoleucine, valine, cystine, cysteine, methionine, threonine, serine, phenylalanine, tyrosine, tryptophan, proline, hydroxyproline, glutamic acid, and aspartic acid.

[0020] The carbohydrates are selected from one or more of glucose, fructose, and sucrose.

[0021] The weight ratio of nitrogen, phosphorus, and potassium in the compound fertilizer is (10 - 20):(10 - 20):(10 - 20), preferably 15:15:15.

[0022] The preparation method of the microbial preparation includes:

[0023] (1) Add Bacillus subtilis culture solution, shrimp and crab shells, collagen, and sodium chloride to water, mix them, ferment at a temperature of 25 - 35°C for 3 - 5 days, and bubble aeration to obtain a fermentation broth;

[0024] (2) Add compound amino acids, carbohydrates, and compound fertilizers to the fermentation broth obtained in step (1), ferment at a temperature of 25 - 35°C for 3 - 5 days, and bubble aeration to obtain a microbial preparation.

[0025] The viable bacteria count in the microbial preparation obtained in step (2) is not less than 1×10 9 ~2×10 10 cfu / ml, preferably the viable bacteria count is not less than 1×10 10 cfu / ml.

[0026] The application method of medium and trace elements and the microbial preparation is to dilute the microbial preparation 1 - 10 times with water to obtain a diluted solution of the microbial preparation, add the above-mentioned medium and trace element composition to the diluted solution of the microbial preparation, mix evenly, and irrigate the roots of Saposhnikovia divaricata. The usage amount of the diluted solution of the microbial preparation is 100L - 1000L per mu.

[0027] Compared with the clear water group, when the medium and trace elements and the microbial preparation of the present invention are jointly applied to the cultivation of Saposhnikovia divaricata, the content of 3'-O-angeloyl-hamaudol can be increased by more than 50%, the contents of prim-O-glucosylcimifugin and 5-O-methylvisammioside can be increased by more than 80%, and the contents of cimifugin and sec-O-glucosylhamaudol can be increased by more than 150%. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is the HPLC chromatogram of the reference solution in Test Example 1 (Peak 1 is prim-O-glucosylcimifugin, Peak 2 is cimifugin, Peak 3 is 5-O-methylvisammioside, Peak 4 is sec-O-glucosylhamaudol, Peak 5 is 3'-O-angeloyl-hamaudol).

[0029] Figure 2 It is the HPLC chromatogram of the test solution in Test Example 1 (Peak 1 is prim-O-glucosylcimifugin, Peak 2 is cimifugin, Peak 3 is 5-O-methylvisammioside, Peak 4 is sec-O-glucosylhamaudol, Peak 5 is 3'-O-angeloyl-hamaudol). DETAILED DESCRIPTION OF THE INVENTION

[0030] Example 1

[0031] Influence of Medium and Trace Elements on the Chromone Content of Saposhnikovia divaricata

[0032] In this study, the orthogonal test method was adopted, and the L 25 (5 6 ) orthogonal test table was selected. Each factor had 5 levels, with a total of 25 tests, 3 replicates for each test, a total of 75 bags. The factor levels are shown in Table 1, and the test design is shown in Table 2.

[0033] Table 1 Orthogonal Test Factor-Level Table (mg / kg)

[0034] Factor level <![CDATA[CaCl2 (A)]]> <![CDATA[MgSO4 (B)]]> <![CDATA[FeCl3 (C)]]> <![CDATA[MnCl2 (D)]]> <![CDATA[CuCl2 (E)]]> <![CDATA[ZnCl2(F)]]> 1 0 0 0 0 0 0 2 300 60 15 10 10 5 3 600 90 30 20 20 10 4 900 120 60 40 40 20 5 1200 180 120 80 80 40

[0035] Table 2 Experimental Design

[0036] Experiment A (mg / kg) B (mg / kg) C (mg / kg) D (mg / kg) E (mg / kg) F (mg / kg) 1 0 0 0 0 0 0 2 0 60 15 10 10 5 3 0 90 30 20 20 10 4 0 120 60 40 40 20 5 0 180 120 80 80 40 6 300 0 15 20 40 40 7 300 60 30 40 80 0 8 300 90 60 80 0 5 9 300 120 120 0 10 10 10 300 180 0 10 20 20 11 600 0 30 80 10 20 12 600 60 60 0 20 40 13 600 90 120 10 40 0 14 600 120 0 20 80 5 15 600 180 15 40 0 10 16 900 0 60 10 80 10 17 900 60 120 20 0 20 18 900 90 0 40 10 40 19 900 120 15 80 20 0 20 900 180 30 0 40 5 21 1200 0 120 40 20 5 22 1200 60 0 80 40 10 23 1200 90 15 0 80 20 24 1200 120 30 10 0 40 25 1200 180 60 20 10 0

[0037] Select plump and uniform Saposhnikovia divaricata seeds, soak them in 35°C water for 24 hours, then take them out and sow them in a petri dish lined with two layers of moist filter paper, and place them at room temperature for germination. After the seed embryos grow to about 1 cm, select seedlings with similar growth vigor and plant them into mushroom bags filled with substrate soil. Add the corresponding contents of medium and trace elements to the substrate soil according to Table 2, mix evenly. Each experiment is divided into 3 bags, a total of 75 bags. Plant 5 plants in each mushroom bag and place them in the greenhouse to grow. Cultivate under the conditions of a daytime temperature of 25°C, a nighttime temperature of 20°C, a humidity of 50%, ventilation for 8 - 12 hours, and a light of 16 hours / darkness of 8 hours. After two true leaves grow, thin out to 3 plants.

[0038] Samples are collected after four months of growth. Wash the roots clean, dry them, and then crush them to measure the chromone content.

[0039] Example 2

[0040] Preparation of Microbial Agent Dilute Solution

[0041] 1 Kg of Bacillus subtilis tkm-1 culture solution, 4 Kg of crab powder, 3 kg of porcine skin collagen, 0.3 kg of sodium chloride, 20 kg of compound amino acids, 10 kg of fructose, 5 kg of compound fertilizer (nitrogen:phosphorus:potassium ratio 15:15:15).

[0042] Bacillus subtilis tkm-1 is sourced from Beijing Tech-Max High-Tech Co., Ltd.

[0043] Preparation method of microbial agent:

[0044] (1) Add Bacillus subtilis culture solution, crab powder, collagen, and sodium chloride to 1000 L of water, and conduct cultivation by bubbling and aeration. Control the cultivation temperature at 28°C and the cultivation time at 4 days to obtain a fermentation broth;

[0045] (2) Add compound amino acids, fructose, and compound fertilizer to the fermentation broth in step (1), and continue to conduct cultivation by bubbling and aeration. Control the cultivation temperature at 32°C and the cultivation time at 3 days to obtain the microbial agent, which is reserved for use.

[0046] Detect the bacterial density of the obtained microbial agent, and the bacterial density is 2.2×10 11 cfu / ml.

[0047] The obtained microbial agent was diluted 10 times with water to obtain a diluted solution of the microbial agent.

[0048] Example 3

[0049] Effects of medium and trace elements and microbial agent on chromone in Saposhnikovia divaricata

[0050] Plump and uniform Saposhnikovia divaricata seeds were selected, soaked in water at 35 °C for 24 hours and then fished out. After being fished out, they were placed on a wet cloth, then placed in a room-temperature environment, covered with a moist hemp sheet to achieve the purpose of germination. After that, the seeds were stirred with sand, and then sown on the ridge surface. After covering the seeds with a gentle sweep of a broom, they were slightly compacted. After the seedlings emerged evenly, in addition to conventional fertilization and watering, 100 seedlings were selected in each group, and root irrigation solution was applied in the manner shown in Table 3 once a month for a total of 3 times.

[0051] Table 3 Root irrigation solution and volume

[0052] Group Root irrigation solution Volume of root irrigation solution Clear water group Clear water 50 ml / plant Group 1 Clear water + Medium and trace element composition 50 ml / plant Group 2 Diluent of Example 2 50 ml / plant Group 3 Diluent of Example 2 + Medium and trace element composition 50 ml / plant

[0053] The composition ratios of the medium and trace element compositions in Group 1 and Group 3 were the same as those in Level 3 of Example 1. The medium and trace elements were added to the diluted solution for root irrigation together. The ratio of the diluted solution to the medium and trace element composition was as follows: The medium and trace elements added to each milliliter of the diluted solution were 72 mg of CaCl2, 10.8 mg of MgSO4, 3.6 mg of FeCl3, 2.4 mg of MnCl2, 2.4 mg of CuCl2, and 1.2 mg of ZnCl2 respectively.

[0054] Test Example 1

[0055] Determination of chromone content in Saposhnikovia divaricata

[0056] Determination method

[0057] Preparation of reference substance solution: Appropriate amounts of prim-O-glucosylcimifugin reference substance, cimifugin reference substance, 5-O-methylvisammioside reference substance, sec-O-glucosylhamaudol reference substance, and 3'-O-angeloylsec-O-glucosylhamaudol reference substance were accurately weighed and dissolved in methanol to prepare a mixed reference substance solution containing 295.08 μg of prim-O-glucosylcimifugin, 76.51 μg of cimifugin, 356.13 μg of 5-O-methylvisammioside, 444.40 μg of sec-O-glucosylhamaudol, and 499.19 μg of 3'-O-angeloylsec-O-glucosylhamaudol in each 1 ml.

[0058] Preparation of test solution: About 1.25 g of the fine powder of this product was accurately weighed and placed in a stoppered conical flask. 50 mL of methanol was accurately added, weighed, refluxed in a water bath for 2 h, cooled, weighed again, and the lost weight was made up with methanol. After shaking well, it was filtered, and the subsequent filtrate was taken to obtain the test solution.

[0059] Detection conditions: Using a GL Sciences-C18 column (4.6 mm × 250 mm, 5 μm) as the chromatographic column, acetonitrile as mobile phase A, and 0.1% phosphoric acid solution as mobile phase B, eluting according to the gradient (0 - 11 min, 15% - 21% A; 11 - 20 min, 21% - 31% A; 20 - 40 min, 31% - 83% A; 40 - 48 min, 83% - 90% A; 48 - 55 min, 90% - 100% A); the detection wavelength is 254 nm, and the flow rate is 1.0 mL·min -1 , and the injection volume is 10 μl.

[0060] The HPLC chromatograms of the reference solution and the test solution are shown in Figure 1 and Figure 2 respectively. Peak 1 is cimifugin, peak 2 is cimifugin, peak 3 is 5-O-methylvisammioside, peak 4 is sec-O-glucosylhamaudol, and peak 5 is 3'-O-angeloylsec-O-glucosylhamaudol. The content of chromone in the test sample is calculated based on the ratio of the peak area of the test sample to that of the reference sample. The larger the peak area ratio, the higher the content of the chromone.

[0061] Determination results of chromone content in the sample of Example 1

[0062] Table 4 Determination results of chromone content in Saposhnikovia divaricata (mg / g)

[0063] Experiment Prim-O-glucosylcimifugin Cimifugin 5-O-methylvisammioside Sec-O-glucosylhamaudol 3’-O-Angeloylsec-O-glucosylhamaudol 1 2.04±0.07 0.22±0.03 2.41±0.12 0.21±0.03 0.15±0.04 2 1.95±0.12 0.37±0.05 2.55±0.23 0.30±0.04 0.20±0.04 3 3.46±0.21 0.27±0.05 3.12±0.15 0.52±0.09 0.35±0.04 4 4.25±0.09 0.70±0.06 3.53±0.22 0.49±0.06 0.23±0.05 5 3.41±0.26 0.68±0.04 3.52±0.18 0.42±0.06 0.21±0.03 6 3.47±0.16 0.38±0.04 3.19±0.17 0.24±0.06 0.23±0.06 7 3.42±0.22 0.53±0.06 3.15±0.20 0.34±0.06 0.23±0.05 8 2.63±0.20 0.65±0.04 2.73±0.15 0.35±0.06 0.28±0.04 9 3.21±0.13 0.29±0.05 2.53±0.19 0.37±0.04 0.28±0.05 10 3.65±0.17 0.42±0.04 3.18±0.16 0.33±0.05 0.23±0.06 11 3.19±0.11 0.62±0.07 2.95±0.11 0.36±0.03 0.20±0.05 12 3.75±0.13 0.46±0.07 2.68±0.13 0.29±0.04 0.30±0.03 13 3.04±0.10 0.31±0.03 3.37±0.38 0.39±0.06 0.29±0.05 14 3.46±0.11 0.45±0.06 3.15±0.25 0.38±0.06 0.25±0.06 15 2.99±0.12 0.38±0.03 3.30±0.33 0.48±0.07 0.19±0.03 16 4.03±0.12 0.59±0.05 2.99±0.15 0.38±0.04 0.24±0.05 17 3.68±0.15 0.48±0.05 3.67±0.31 0.39±0.04 0.31±0.02 18 2.79±0.24 0.36±0.04 3.35±0.24 0.34±0.04 0.26±0.04 19 3.23±0.06 0.35±0.07 2.94±0.14 0.45±0.07 0.35±0.07 20 4.13±0.16 0.52±0.05 3.27±0.27 0.43±0.06 0.34±0.06 21 2.78±0.25 0.58±0.07 3.41±0.24 0.29±0.03 0.37±0.05 22 2.78±0.22 0.23±0.05 2.99±0.12 0.32±0.06 0.31±0.04 23 3.06±0.28 0.74±0.08 3.15±0.19 0.40±0.06 0.26±0.05 24 3.52±0.20 0.22±0.04 3.05±0.24 0.40±0.05 0.30±0.07 25 2.90±0.23 0.19±0.03 2.94±0.18 0.38±0.03 0.28±0.07

[0064] Performing an analysis of variance on the above results, the results are shown in Table 5

[0065] The influence of trace elements on the chromone content in Table 5

[0066] Element Prim-O-glucosylcimifugin Cimifugin 5-O-methylvisammioside Sec-O-glucosylhamaudol 3’-O-Angeloylsec-O-glucosylhamaudol <![CDATA[CaCl2 (A)]]> P＜0.01 P＜0.01 P＜0.05 P＜0.01 P＜0.01 <![CDATA[MgSO4 (B)]]> P＜0.01 P＜0.01 P＜0.01 P＜0.01 P＜0.05 <![CDATA[FeCl3 (C)]]> P＜0.01 P＜0.01 P＜0.01 P＜0.01 P＞0.05 <![CDATA[MnCl2 (D)]]> P＜0.01 P＜0.01 P＜0.01 P＞0.05 P＞0.05 <![CDATA[CuCl2 (E)]]> P＜0.01 P＜0.01 P＜0.05 P＞0.05 P＜0.01 <![CDATA[ZnCl2(F)]]> P＜0.01 P＜0.01 P＜0.01 P＜0.05 P＞0.05

[0067] The results show that medium and trace elements have significant effects on the contents of cimifugin, cimifugin, and 5-O-methylvisammioside. The effects of MnCl2 (D) and CuCl2 (E) elements on sec-O-glucosylhamaudol are not significant, and the effects of FeCl3 (C), MnCl2 (D), and ZnCl2 (F) elements on 3'-O-angeloylsec-O-glucosylhamaudol are not significant.

[0068] Determination results of chromone content in the sample of Example 3

[0069] Table 6 Determination results of chromone content in Saposhnikovia divaricata (mg / g)

[0070] Group Prim-O-glucosylcimifugin Cimifugin 5-O-methylvisammioside Sec-O-glucosylhamaudol 3’-O-Angeloylsec-O-glucosylhamaudol Clear water group 2.66 0.37 2.23 0.22 0.33 Group 1 3.7 0.52 2.98 0.30 0.42 Group 2 3.36 0.68 3.34 0.41 0.40 Group 3 5.07 0.95 4.12 0.63 0.52

[0071] Compared with the water group, the percentage increase in the active ingredients is shown in Table 7:

[0072] Table 7 Percentage increase in active ingredients compared to the pure water group (%)

[0073] Group Prim-O-glucosylcimifugin Cimifugin 5-O-methylvisammioside Sec-O-glucosylhamaudol 3’-O-Angeloylsec-O-glucosylhamaudol (Group 1 - Clear water group) / Clear water group 39.10 40.54 33.63 36.36 27.27 (Group 2 - Clear water group) / Clear water group 26.32 83.78 49.78 86.36 21.21 (Group 3 - Clear water group) / Clear water group 90.60 156.76 84.75 186.36 57.58

[0074] As can be seen from Table 7, compared to the pure water group, the content of active ingredients in Saposhnikovia divaricata in Groups 1 - 3 was significantly higher. Compared to Groups 1 and 2, Group 3 could significantly increase the content of active ingredients in Saposhnikovia divaricata.

[0075] The above content describes in detail the preferred embodiments of the present invention. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention. These simple modifications all fall within the protection scope of the present invention.

[0076] In addition, it should be noted that the various specific technical features described in the above specific embodiments can be combined in any way without conflict. To avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

[0077] Furthermore, any combination can be made between various different embodiments of the present invention as long as it does not violate the idea of the present invention, and it should also be regarded as the content disclosed by the present invention.

Claims

1. A planting method for increasing the chromone content in Saposhnikovia divaricata, characterized in that, The combined application of the medium and trace element composition and the microbial agent in the cultivation of Saposhnikovia divaricata. The strain in the microbial agent is Bacillus subtilis, and the chromones are prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside, sec-O-glucosylhamaudol, and 3'-O-angeloylsec-O-glucosylhamaudol. The Bacillus subtilis is tkm-1, with the preservation number of CGMCC No. 14950. The Bacillus subtilis is inoculated into the LB medium and cultured at 30-32 °C for 24-48 h to obtain the Bacillus subtilis culture solution. The microbial agent also includes shrimp and crab shells, collagen, sodium chloride, compound amino acids, carbohydrates, and compound fertilizers. The weight ratio of the Bacillus subtilis culture solution to shrimp and crab shells, collagen, sodium chloride, compound amino acids, carbohydrates, and compound fertilizers is 1:(1-10):(1-10):(0.1-1):(10-50):(1-20):(1-10). The microbial agent is prepared by the following steps: (1) Add the Bacillus subtilis culture solution, shrimp and crab shells, collagen, and sodium chloride into water, mix them, ferment at 25-35 °C for 3-5 days, and bubble and ventilate to obtain the fermentation broth; (2) Add the compound amino acids, carbohydrates, and compound fertilizers into the fermentation broth obtained in step (1), ferment at 25-35 °C for 3-5 days, and bubble and ventilate to obtain the product.

2. The planting method according to claim 1, characterized in that, The medium and trace element composition is CaCl2, MgSO4, FeCl3, MnCl2, CuCl2, ZnCl2.

3. The planting method according to claim 2, characterized in that, In the medium and trace element composition, CaCl2 is 300-1200 parts by weight, MgSO4 is 60-180 parts by weight, FeCl3 is 15-120 parts by weight, MnCl2 is 10-80 parts by weight, CuCl2 is 10-80 parts by weight, and ZnCl2 is 5-40 parts by weight.

4. The planting method according to claim 3, characterized in that, In the medium and trace element composition, CaCl2 is 300-900 parts by weight, MgSO4 is 60-120 parts by weight, FeCl3 is 15-60 parts by weight, MnCl2 is 10-40 parts by weight, CuCl2 is 10-40 parts by weight, and ZnCl2 is 5-20 parts by weight.

5. The planting method according to claim 4, characterized in that In the medium and trace element composition, CaCl2 is 600 parts by weight, MgSO4 is 90 parts by weight, FeCl3 is 30 parts by weight, MnCl2 is 20 parts by weight, CuCl2 is 20 parts by weight, and ZnCl2 is 10 parts by weight.

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