Search

CN-122004129-A - Culture medium of barbed skullcap herb and preparation method thereof

CN122004129ACN 122004129 ACN122004129 ACN 122004129ACN-122004129-A

Abstract

The invention relates to the technical field of barbed skullcap herb culture, in particular to a barbed skullcap herb culture medium and a preparation method thereof, wherein the barbed skullcap herb culture medium is prepared from an MS culture medium and basic raw materials according to a mass ratio of 6-7:1, and the basic raw materials comprise, by weight, 2-4 parts of novel nano silicon dioxide, 0.05-0.08 part of cerium nitrate, 1-2 parts of organic microbial inoculum, 0.7-0.8 part of 6-benzyl adenine, 0.2-0.4 part of alpha-naphthylacetic acid, 0.1-0.3 part of methyl jasmonate and 2-4 parts of functional auxiliary agents. The invention enhances the stability of cell walls and realizes the slow release regulation of hormone by novel nano silicon dioxide, and improves the cerium nitrate to activate flavonoid compound synthase system, the organic microbial inoculum inhibits pathogenic bacteria and promotes nitrogen conversion by means of the exocrine lipopeptide antibiotics and the nitrogen fixation enzyme system, and the functional auxiliary agent realizes the efficient retention and stable release of active ingredients.

Inventors

  • HU WEIQI
  • LI YU
  • ZHANG QIAOLI

Assignees

  • 广州营发科技有限公司

Dates

Publication Date
20260512
Application Date
20260120

Claims (10)

  1. 1. A culture medium of barbed skullcap herb is characterized by being prepared from an MS culture medium and basic raw materials according to a mass ratio of 6-7:1, wherein the basic raw materials comprise, by weight, 2-4 parts of novel nano silicon dioxide, 0.05-0.08 part of cerium nitrate, 1-2 parts of an organic microbial inoculum, 0.7-0.8 part of 6-benzyladenine, 0.2-0.4 part of alpha-naphthylacetic acid, 0.1-0.3 part of methyl jasmonate and 2-4 parts of a functional auxiliary agent.
  2. 2. The culture medium of the sculellaria barbata according to claim 1, wherein the preparation method of the functional auxiliary agent comprises the following steps: S1, cleaning the barbed skullcap herb by deionized water, draining, drying in a 35-40 ℃ hot air drying oven until the water content is less than or equal to 5%, crushing, and sieving with a 50-60 mesh sieve to obtain barbed skullcap herb powder; S2, preparing an extraction solvent, mixing the barbed skullcap herb powder with the extraction solvent according to the mass ratio of 1:18-22, placing the mixture in an ultrasonic extractor at 45-50 ℃ for 30-35 min to obtain a crude extract, centrifuging the crude extract, collecting a supernatant and filtering to obtain the extract; S3, mixing the extracting solution, cysteine and disodium ethylenediamine tetraacetate according to the mass ratio of 1:0.0008-0.012:0.0002-0.0004, stirring for 25-35 min at 23-27 ℃ to form an inhibiting extracting solution, mixing the inhibiting extracting solution, ascorbic acid, glutathione and citric acid according to the mass ratio of 1:0.0002-0.0004:0.0008-0.0012:0.0004-0.0006, and adding sodium hydroxide solution to adjust the pH value to 5.0-5.2 to obtain an antioxidation treatment solution; and S4, filtering and purifying the antioxidant treatment liquid to obtain a clarified liquid, and treating the clarified liquid to prepare the functional auxiliary agent.
  3. 3. The culture medium of Scutellaria barbata according to claim 2, wherein the extraction solvent in the step S2 is prepared by the following method: respectively preparing a citric acid solution and a disodium hydrogen phosphate solution, mixing the citric acid solution and the disodium hydrogen phosphate solution according to a volume ratio of 1:1.2-1.4 to obtain a buffer solution, dissolving ascorbic acid in the buffer solution according to a solid-to-liquid ratio of 0.0015-0.002g/mL, and stirring for 10-15 min; And (3) introducing nitrogen into the buffer solution containing the ascorbic acid, and stirring for 25-35 min to obtain the extraction solvent.
  4. 4. The culture medium of Scutellaria barbata according to claim 2, wherein the antioxidant treatment solution is filtered and purified to obtain a clarified solution, and the specific steps are as follows: Filtering the oxidation resisting treatment solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 3kDa, collecting the ultrafiltrate, adsorbing by using a macroporous adsorption resin column, eluting by using 70% ethanol after the adsorption is saturated to form an eluent, and evaporating the eluent to one fifth of the original volume to obtain a concentrated solution; adding active carbon into the concentrated solution according to the solid-liquid ratio of 0.0015-0.002g/mL, stirring in a water bath at 50-55 ℃ for 40-50 min, and filtering after stirring to obtain a clear liquid.
  5. 5. The culture medium of Scutellaria barbata according to claim 2, wherein the clarified liquid is further treated by the following steps: mixing the clarified liquid and beta-cyclodextrin according to a mass ratio of 4.5-5.5:1, stirring in a water bath at 40-45 ℃ for 1.5-2.5 hours to form an inclusion compound solution, and conveying the inclusion compound solution to a spray dryer to obtain microcapsule powder; Mixing gelatin and Arabic gum according to a mass ratio of 1:1, dissolving the mixture in deionized water to prepare a wall material solution with a concentration of 8% -10%, mixing microcapsule powder and the wall material solution according to a mass ratio of 1:4.8% -5.2, stirring the mixture at 55-65 ℃ to form an emulsion, adjusting the pH of the emulsion to 4.4-4.6, adding transglutaminase with a mass of 0.4% -0.6% of the wall material solution after the adjustment is completed, reacting for 0.8-1 h at 35-45 ℃, after the reaction is completed, dripping a calcium chloride solution with a mass fraction of 5%, preparing a microcapsule, and washing the microcapsule for 2-4 times by using deionized water to obtain the functional auxiliary agent.
  6. 6. The culture medium of the sculellaria barbata according to claim 3, wherein the citric acid solution is prepared by mixing citric acid and deionized water according to a mass ratio of 1:35-40; the disodium hydrogen phosphate solution is prepared by mixing disodium hydrogen phosphate and deionized water according to a mass ratio of 1:20-24.
  7. 7. A culture medium of sculellaria barbata according to claim 1, wherein the novel nano-silica is prepared by the steps of: Mixing nano silicon dioxide with the particle size of 20nm, absolute ethyl alcohol and 3-aminopropyl triethoxysilane according to the mass ratio of 6-7:13-15:1, stirring for 5.5-6.5 hours under the reflux condition of 65-75 ℃ to obtain a reaction liquid, centrifuging and washing 2-4 times by using ethanol after the reaction liquid is cooled to 25 ℃, and drying for 10-12 hours under the vacuum environment of 55-65 ℃ to obtain modified silicon dioxide; Grafting the modified silicon dioxide to obtain grafted silicon dioxide, mixing the grafted silicon dioxide, phosphate buffer solution and 6-benzyl adenine according to the mass ratio of 23-27:80-85:1, stirring for 20-25 h at 23-27 ℃ to obtain a load solution, dialyzing the load solution, collecting a dialyzate, and freeze-drying to obtain the novel nano silicon dioxide.
  8. 8. The culture medium of Scutellaria barbata according to claim 7, wherein the grafting of the modified silica comprises the following steps: Mixing modified silicon dioxide, deionized water and polyethylene glycol according to a mass ratio of 1:2.5-3.5:1.5-2.5 to obtain a polymer solution, adding ammonium persulfate with a mass of 0.4-0.6% of the polyethylene glycol into the polymer solution, and stirring for 7.5-8.5 hours at 55-65 ℃ under the protection of nitrogen to form a grafting solution; Dialyzing the grafting solution with deionized water, and freeze-drying after the dialysis is finished to obtain the grafted silicon dioxide.
  9. 9. The culture medium of Scutellaria barbata according to claim 1, wherein the organic microbial inoculum is any one of Bacillus subtilis, bacillus bailii and Bacillus amyloliquefaciens.
  10. 10. The preparation method of the culture medium of the sculellaria barbata according to any one of claims 1 to 9, which is characterized by comprising the following steps: Weighing each salt according to a standard formula of an MS culture medium, dissolving in deionized water, heating and dissolving, adding sucrose and agar, adjusting the pH to 5.8-6, and sterilizing at a high pressure of 121-125 ℃ for 15-20 min; Step two, activating the organic bacterial to obtain an organic bacterial agent, preparing a functional auxiliary agent and novel nano silicon dioxide according to the steps, respectively adding the novel silicon dioxide, cerium nitrate, the organic bacterial agent, 6-benzyl adenine, alpha-naphthylacetic acid, methyl jasmonate and the functional auxiliary agent after cooling and solidifying the MS culture medium to 45 ℃, and obtaining the culture medium of the sculellaria barbata.

Description

Culture medium of barbed skullcap herb and preparation method thereof Technical Field The invention relates to the technical field of barbed skullcap herb culture, in particular to a barbed skullcap herb culture medium and a preparation method thereof. Background The herba Scutellariae Barbatae is a herb plant of the genus herba Scutellariae Barbatae of the family Labiatae, which generally grows in warm and humid environments and is widely distributed in Europe, asia and North America, has low plant content, generally has opposite leaves and purple or white flowers, has compact inflorescence, has a lip-shaped corolla, has certain ornamental value in gardening, is applied to the field of Chinese herbal medicines, has certain medicinal value, and is commonly used for clearing heat, detoxicating, promoting urination, reducing swelling and the like. The conventional barbed skullcap herb culture medium has the problems that firstly, the conventional culture medium only depends on basic nutrient salt and single hormone combination, so that the growth efficiency is low, the accumulation of active ingredients is insufficient, the growth of plants and the improvement of active ingredients are difficult to achieve, secondly, the conventional nano material is easy to agglomerate and settle in the culture medium, so that the bioavailability is low, even toxicity is possibly caused to cells, the culture effect is influenced, the conventional system is too serious in bud differentiation and rooting, the regulation of adversity stress and the induction of secondary metabolites are neglected, the plant stress resistance is insufficient, the active ingredient synthesis efficiency is low, and finally, the browning problem in the barbed skullcap herb tissue culture is serious, and the conventional method is not obvious due to the fact that the activity of polyphenol oxidase is too high, so that the brown change rate is easy to be high, and the retention of the active ingredients and the plant growth health are influenced. Accordingly, the present invention provides a culture medium of Scutellaria barbata and a preparation method thereof, which are used for solving the above-mentioned related technical problems. Disclosure of Invention The invention aims to provide a culture medium of barbed skullcap herb and a preparation method thereof, wherein an MS culture medium is taken as a basic nutrition carrier, the stability of a cell wall is enhanced and hormone slow-release regulation and control are realized through novel nano silicon dioxide, cerium nitrate is improved to activate a flavonoid compound synthase system, an organic microbial agent is used for inhibiting pathogenic bacteria and promoting nitrogen conversion through a lipo-peptide antibiotic and a nitrogen fixation enzyme system, 6-benzyl adenine and alpha-naphthylacetic acid are combined to cooperate with a novel methyl jasmonate hormone, so that bud differentiation and rooting are promoted, stress resistance and accumulation of secondary metabolites are induced, and functional auxiliaries are used for realizing efficient retention and stable release of active ingredients. In order to achieve the above purpose, the present invention provides the following technical solutions: the culture medium of the sculellaria barbata is prepared from an MS culture medium and basic raw materials according to a mass ratio of 6-7:1, wherein the basic raw materials comprise, by weight, 2-4 parts of novel nano silicon dioxide, 0.05-0.08 part of cerium nitrate, 1-2 parts of an organic microbial inoculum, 0.7-0.8 part of 6-benzyl adenine, 0.2-0.4 part of alpha-naphthylacetic acid, 0.1-0.3 part of methyl jasmonate and 2-4 parts of a functional auxiliary agent. Preferably, 6-benzyl adenine is purchased from Shandong Hengke Biotechnology Co., ltd, whereas cerium nitrate is selected from Shandong Liang New Material technologies Co., ltd, and α -naphthalene acetic acid is purchased from Beijing Washingchao, methyl jasmonate is selected from Hubei Yamaide biological medicine Co., ltd. Further, as shown in fig. 1, the preparation method of the functional auxiliary agent comprises the following steps: S1, cleaning the barbed skullcap herb by deionized water, draining, drying in a 35-40 ℃ hot air drying oven until the water content is less than or equal to 5%, crushing, and sieving with a 50-60 mesh sieve to obtain barbed skullcap herb powder; S2, preparing an extraction solvent, mixing the barbed skullcap herb powder with the extraction solvent according to the mass ratio of 1:18-22, placing the mixture in an ultrasonic extractor at 45-50 ℃ for 30-35 min to obtain a crude extract, centrifuging the crude extract, collecting a supernatant and filtering to obtain the extract; S3, mixing the extracting solution, cysteine and disodium ethylenediamine tetraacetate according to the mass ratio of 1:0.0008-0.012:0.0002-0.0004, stirring for 25-35 min at 23-27 ℃ to form an inhibiting extracting solution,