A north chicken head rhizome bud induction medium and preparation and culture method

Abstract

The application discloses a kind of northern chicken head rhizome bud induction medium and its preparation and culture method, belong to medicinal plant tissue culture technical field.The medium is based on MS, contains 6-BA 0.5~3.5 mg, NAA 0.1~2.0 mg, PVP 100~1500 mg, Vc 20~150 mg, sucrose 30 g, carrageenan 7 g per liter, pH 5.8.Its preparation method includes: S1 dissolves constant volume, S2 adds hormone and antioxidant, S3 adjusts pH, S4 sterilization, S5 is cooled to 95 ℃ and is shaken to eliminate condensate water after cooling solidification.Culture method is: after disinfection, rhizome bud explant is inoculated on the medium, is cultured under the condition of 24±2 ℃, 2000 lx, 12h light and dark alternation and induces adventitious bud.The application is brown by PVP and Vc synergistic, optimizes hormone ratio, carrageenan stabilizes solidification and is treated after sterilization and eliminates condensate water, effectively reduces browning rate, shortens induction period, improves the number of shoots, applicable to factory seedling raising.

Description

Technical Field

[0001] This invention relates to the field of medicinal plant tissue culture technology, specifically to a root and stem bud induction culture medium for Polygonatum odoratum from northern China and its preparation and culture method. Background Technology

[0002] Plant tissue culture is an effective technique for the large-scale rapid propagation of Polygonatum odoratum seedlings. Currently, publicly available research on Polygonatum odoratum tissue culture and its culture medium formulations in China mainly focuses on southern varieties such as Polygonatum multiflorum and Polygonatum yunnanense. Regarding culture medium preparation, MS medium is the most commonly used basal medium, with sucrose often used as the carbon source and agar as the solidifying agent. Plant growth regulators typically include 6-benzylaminopurine and naphthaleneacetic acid, but the specific dosage varies depending on the variety and culture stage, and there is no unified standard. Regarding browning suppression measures, some studies have attempted to add a single antioxidant to the culture medium to reduce the browning rate of explants. Furthermore, the routine procedures for tissue culture in existing technologies generally include basic steps such as culture medium preparation, pH adjustment, high-temperature sterilization, cooling and solidification, explant disinfection and inoculation, and constant temperature and light culture.

[0003] However, existing technologies still have the following shortcomings: The polyphenol content in the rhizomes and buds of *Polygonatum sibiricum* from northern China is high. When conventional MS medium and formulations for southern varieties are directly applied, a single antioxidant system is insufficient to effectively inhibit explant browning, leading to extensive necrosis of the inoculated material. Current hormone formulations are mostly fixed concentration combinations, lacking precise and synergistic regulation for the differentiation of *Polygonatum sibiricum* rhizomes and buds, easily resulting in problems such as excessive callus, insufficient adventitious bud differentiation, and bud malformation. Traditional agar solidifying agents are prone to liquefaction during culture, increasing the risk of contamination. Furthermore, the condensation generated on the inner wall of the flask during the cooling stage after high-temperature sterilization of the medium is often overlooked, causing explants to rot due to prolonged soaking. These defects combine to result in a long adventitious bud induction period, a low number of buds, and a low success rate, severely restricting the industrial application of rapid propagation technology for *Polygonatum sibiricum* tissue culture.

[0004] Existing technologies lack a primary induction culture medium specifically for the root, stem, and bud of Polygonatum odoratum from northern China. Single antioxidants are insufficient to inhibit browning, hormone ratios are unsuitable, coagulants are prone to liquefaction, condensation water causes explant rot, and the induction period is long with few buds. Summary of the Invention

[0005] (a) Technical problems to be solved To address the shortcomings of existing technologies, this invention provides a root and stem bud induction culture medium for *Polygonatum sibiricum* from northern China, along with its preparation and cultivation method. The medium is based on MS medium, supplemented with 0.5–3.5 mg / L 6-BA, 0.1–2.0 mg / L NAA, 100–1500 mg / L PVP, 20–150 mg / L Vc, 30 g / L sucrose, and 7 g / L carrageenan, at pH 5.8. After dissolving, adding hormones, adjusting pH, and sterilizing, the medium is shaken clockwise at 95°C to remove condensation and cooled for later use. The sterilized root and stem bud explants are inoculated onto this medium and cultured at 24±2°C, 2000 lx, under alternating light and dark conditions for 12 hours to induce adventitious buds.

[0006] (II) Technical Solution To achieve the above objectives, the present invention provides the following technical solution: a root and stem bud induction culture medium for *Polygonatum sibiricum* from northern China and its preparation and cultivation method, comprising using MS medium as a base, wherein each liter of medium contains: 0.5–3.5 mg of 6-benzylaminopurine, 0.1–2.0 mg of naphthaleneacetic acid, 100–1500 mg of polyvinylpyrrolidone, 20–150 mg of L-ascorbic acid, 30 g of sucrose, and 7 g of carrageenan, and is diluted to 1 L with distilled water; the pH of the culture medium is 5.8.

[0007] Preferably, the culture medium contains 2.5 mg of 6-benzylaminopurine, 0.8 mg of naphthaleneacetic acid, 800 mg of polyvinylpyrrolidone, and 80 mg of L-ascorbic acid per liter.

[0008] Preferably, the culture medium is used for the primary induction culture of explants from the rhizome buds of Polygonatum odoratum from northern China.

[0009] Preferably, it includes the following steps: S1: Dissolve MS dry powder, sucrose and carrageenan in distilled water and bring the volume to 1L; S2: Add 6-benzylaminopurine, naphthaleneacetic acid, polyvinylpyrrolidone and L-ascorbic acid, and mix well; S3: Adjust pH to 5.8; S4: After dispensing, sterilize at 121℃ for 20 minutes; S5: After sterilization, cool to 95℃, remove the culture bottle, shake it clockwise once to spread the culture medium on the inner wall of the bottle, and then lay it flat to cool and solidify.

[0010] Preferably, the shaking time in step S5 is 3 to 5 seconds.

[0011] Preferably, the sterilized rhizome shoot explants are inoculated onto the culture medium described in any one of claims 1 to 3 and cultured at 24±2℃, light intensity of 2000 lx, and a photoperiod of 12h light / 12h darkness.

[0012] Preferably, the disinfection includes treatment with 75% alcohol for 30 seconds and treatment with 0.1% sodium hypochlorite for 8 minutes.

[0013] Preferably, the culture time is 10 to 28 days.

[0014] Preferably, the concentrations are as follows: 6-benzylaminopurine is 2.0–3.0 mg / L, naphthaleneacetic acid is 0.5–1.5 mg / L, polyvinylpyrrolidone is 500–1200 mg / L, and L-ascorbic acid is 50–120 mg / L.

[0015] Preferably, the shaking operation in step S5 is performed while the culture medium is in a molten state.

[0016] Compared with the prior art, the present invention provides a culture medium for inducing rhizome buds of Polygonatum odoratum from northern China, as well as its preparation and cultivation method, which has the following beneficial effects: 1. The present invention adds polyvinylpyrrolidone and L-ascorbic acid to the culture medium at the same time. The two work together to adsorb polyphenols and scavenge free radicals, effectively inhibiting explant browning.

[0017] 2. This invention precisely regulates the balance between cell division and elongation by limiting the synergistic ratio of 6-benzylaminopurine and naphthaleneacetic acid, thereby significantly improving the adventitious bud induction rate and the number of buds.

[0018] 3. This invention uses carrageenan instead of agar as a coagulant, resulting in strong coagulation stability of the culture medium, which is less prone to liquefaction after sterilization, thus reducing the risk of contamination.

[0019] 4. In this invention, after sterilization and cooling to 95°C, the culture bottle is shaken clockwise to allow the molten culture medium to coat the inner wall of the bottle and absorb condensate, effectively eliminating water accumulation inside the bottle and preventing explant rot.

[0020] 5. This invention provides a complete set of processes from culture medium preparation to adventitious bud induction, with standardized operation, suitable for industrialized mass seedling production. Attached Figure Description

[0021] Figure 1 This is a schematic diagram of the method steps of the present invention. Detailed Implementation

[0022] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0023] This invention provides a culture medium for inducing root and stem buds of Polygonatum odoratum from northern China, as well as its preparation and cultivation method. Please refer to [link to relevant documentation]. Figure 1The method includes the following steps: S1: Dissolve MS dry powder, sucrose and carrageenan in distilled water and bring the volume to 1L.

[0024] S2: Add 6-benzylaminopurine, naphthaleneacetic acid, polyvinylpyrrolidone and L-ascorbic acid, and mix well.

[0025] S3: Adjust pH to 5.8.

[0026] S4: After dispensing, sterilize at 121℃ for 20 minutes.

[0027] S5: After sterilization, cool to 95℃, remove the culture bottle, shake it clockwise once to spread the culture medium on the inner wall of the bottle, and then lay it flat to cool and solidify.

[0028] Explant treatment and culture conditions: Newly sprouted rhizomes and shoots from three-year-old, disease-free *Polygonatum sibiricum* were selected as explants. After rinsing the rhizomes and shoots thoroughly with running water, they were sterilized in a clean bench: first, treated with 75% ethanol for 30 seconds, rinsed once with sterile water, then treated with 0.1% sodium hypochlorite for 8 minutes, and rinsed five times with sterile water. After blotting off surface moisture with sterile filter paper, the sterilized rhizome and shoot explants were inoculated onto the aforementioned culture medium and cultured at 24±2℃, light intensity of 2000 lx, and a photoperiod of 12h light / 12h dark.

[0029] Example 1: The purpose of this embodiment is to verify the actual effect of the culture medium under the optimal ratio conditions of the present invention on the induction of adventitious buds from the rhizome of Polygonatum odoratum in northern China.

[0030] The specific implementation steps of this embodiment are as follows: Prepare the culture medium according to the aforementioned method. Each liter of medium contains MS dry powder, 2.5 mg of 6-benzylaminopurine, 0.8 mg of naphthaleneacetic acid, 800 mg of polyvinylpyrrolidone, 80 mg of L-ascorbic acid, 30 g of sucrose, and 7 g of carrageenan. Adjust the pH to 5.8. S1: Dissolve the MS dry powder, sucrose, and carrageenan in distilled water and bring the volume to 1 L. S2: Add 2.5 mg of 6-benzylaminopurine, 0.8 mg of naphthaleneacetic acid, 800 mg of polyvinylpyrrolidone, and 80 mg of L-ascorbic acid, and mix well. S3: Adjust the pH to 5.8. S4: After dispensing, sterilize at 121℃ for 20 min. S5: After sterilization, cool to 95℃, remove the culture flasks, shake clockwise once to coat the inner wall of the flasks with the culture medium, and then lay flat to cool and solidify.

[0031] Newly sprouted rhizomes and buds from three-year-old, disease-free *Polygonatum sibiricum* plants were selected as explants. After rinsing the rhizomes and buds thoroughly with running water, they underwent sterilization in a clean bench: first, treatment with 75% alcohol for 30 seconds, followed by rinsing once with sterile water; then, treatment with 0.1% sodium hypochlorite for 8 minutes, followed by rinsing five times with sterile water. After blotting the surface moisture with sterile filter paper, the sterilized rhizomes and bud explants were inoculated onto the aforementioned culture medium and cultured at 24±2℃, with a light intensity of 2000 lx and a photoperiod of 12h light / 12h darkness.

[0032] The implementation results of this embodiment are as follows: After 15 days of culture, the browning rate of explants was 8.7%, the induction rate of adventitious buds was 92.3%, and an average of 5.2 buds were produced per explant. The induced adventitious buds were robust, green, and vigorous, without any deformities or vitrification.

[0033] Example 2 The purpose of this embodiment is to verify the effectiveness of the culture medium in inducing adventitious buds from the rhizomes of Polygonatum odoratum in northern China under the lower concentration conditions of the formulation of the present invention.

[0034] The specific implementation steps of this embodiment are as follows: Prepare the culture medium according to the aforementioned method. Each liter of medium contains MS dry powder, 0.5 mg of 6-benzylaminopurine, 0.1 mg of naphthaleneacetic acid, 100 mg of polyvinylpyrrolidone, 20 mg of L-ascorbic acid, 30 g of sucrose, and 7 g of carrageenan. Adjust the pH to 5.8. S1: Dissolve the MS dry powder, sucrose, and carrageenan in distilled water and bring the volume to 1 L. S2: Add 0.5 mg of 6-benzylaminopurine, 0.1 mg of naphthaleneacetic acid, 100 mg of polyvinylpyrrolidone, and 20 mg of L-ascorbic acid, and mix well. S3: Adjust the pH to 5.8. S4: Dispense the medium and sterilize at 121°C for 20 min. S5: After sterilization, cool to 95°C, remove the culture flasks, shake clockwise once to coat the inner wall of the flasks with the culture medium, and then lay them flat to cool and solidify.

[0035] Newly sprouted rhizomes and buds from three-year-old, disease-free *Polygonatum sibiricum* plants were selected as explants. After rinsing the rhizomes and buds thoroughly with running water, they underwent sterilization in a clean bench: first, treatment with 75% alcohol for 30 seconds, followed by rinsing once with sterile water; then, treatment with 0.1% sodium hypochlorite for 8 minutes, followed by rinsing five times with sterile water. After blotting the surface moisture with sterile filter paper, the sterilized rhizomes and bud explants were inoculated onto the aforementioned culture medium and cultured at 24±2℃, with a light intensity of 2000 lx and a photoperiod of 12h light / 12h darkness.

[0036] The implementation results of this embodiment are as follows: After 28 days of culture, the browning rate of explants was 13.9%, the adventitious bud induction rate was 82.3%, and an average of 3.1 buds emerged from a single explant. The adventitious buds grew normally, with light green color and good growth.

[0037] Example 3 The purpose of this embodiment is to verify the effectiveness of the culture medium in inducing adventitious buds from the rhizomes of Polygonatum odoratum in northern China under the upper limit concentration conditions of the formulation of the present invention.

[0038] The specific implementation steps of this embodiment are as follows: Prepare the culture medium according to the aforementioned method. Each liter of medium contains MS dry powder, 3.5 mg of 6-benzylaminopurine, 2.0 mg of naphthaleneacetic acid, 1500 mg of polyvinylpyrrolidone, 150 mg of L-ascorbic acid, 30 g of sucrose, and 7 g of carrageenan. Adjust the pH to 5.8. S1: Dissolve the MS dry powder, sucrose, and carrageenan in distilled water and bring the volume to 1 L. S2: Add 3.5 mg of 6-benzylaminopurine, 2.0 mg of naphthaleneacetic acid, 1500 mg of polyvinylpyrrolidone, and 150 mg of L-ascorbic acid, and mix well. S3: Adjust the pH to 5.8. S4: After dispensing, sterilize at 121℃ for 20 min. S5: After sterilization, cool to 95℃, remove the culture flasks, shake clockwise once to coat the inner wall of the flasks with the culture medium, and then lay flat to cool and solidify.

[0039] Newly sprouted rhizomes and buds from three-year-old, disease-free *Polygonatum sibiricum* plants were selected as explants. After rinsing the rhizomes and buds thoroughly with running water, they underwent sterilization in a clean bench: first, treatment with 75% alcohol for 30 seconds, followed by rinsing once with sterile water; then, treatment with 0.1% sodium hypochlorite for 8 minutes, followed by rinsing five times with sterile water. After blotting the surface moisture with sterile filter paper, the sterilized rhizomes and bud explants were inoculated onto the aforementioned culture medium and cultured at 24±2℃, with a light intensity of 2000 lx and a photoperiod of 12h light / 12h darkness.

[0040] The implementation results of this embodiment are as follows: After 22 days of culture, the browning rate of explants was 7.6%, the adventitious bud induction rate was 91.5%, and an average of 4.5 buds emerged from a single explant. The adventitious buds were slightly clustered and robust, and subsequent subculture was not affected.

[0041] Comparative Example Comparative Example 1: The purpose of this comparative study is to verify the effect of conventional MS medium (without a compound antioxidant system) on the induction of adventitious buds from the rhizomes of Polygonatum odoratum in northern China, and to compare it with the technical solution of this invention.

[0042] The specific implementation steps for this comparative example are as follows: Prepare standard MS medium, containing MS powder, 1.0 mg 6-benzylaminopurine, 0.2 mg naphthaleneacetic acid, 30 g sucrose, and 7 g agar per liter, adjusting the pH to 5.8. Dissolve the MS powder, sucrose, and agar in distilled water, bringing the volume to 1 L. Add 1.0 mg 6-benzylaminopurine and 0.2 mg naphthaleneacetic acid, mix well, adjust the pH to 5.8, dispense into flasks, and sterilize at 121°C for 20 min. After sterilization, remove the culture flasks and allow them to cool and solidify horizontally before use. Do not shake clockwise to remove condensate.

[0043] Newly sprouted rhizomes and buds from three-year-old, disease-free *Polygonatum sibiricum* plants were selected as explants. After rinsing the rhizomes and buds thoroughly with running water, they underwent sterilization in a clean bench: first, treatment with 75% alcohol for 30 seconds, followed by rinsing once with sterile water; then, treatment with 0.1% sodium hypochlorite for 8 minutes, followed by rinsing five times with sterile water. After blotting the surface moisture with sterile filter paper, the sterilized rhizomes and bud explants were inoculated onto the aforementioned culture medium and cultured at 24±2℃, with a light intensity of 2000 lx and a photoperiod of 12h light / 12h darkness.

[0044] The results of this comparative study are as follows: After 38 days of culture, the browning rate of explants was 66.2%, and the adventitious bud induction rate was only 35.7%. A large number of explants underwent browning and necrosis, and the surviving buds were weak, yellow, and grew slowly. Obvious condensation droplets were visible on the inner wall of the culture bottle, and some explants showed signs of rotting at their base.

[0045] Comparative Example 2: The purpose of this comparative study is to verify the effect of culture medium with only the single antioxidant PVP added without L-ascorbic acid on the induction of adventitious buds from the rhizomes of Polygonatum odoratum in northern China, and to compare it with the composite antioxidant system of this invention.

[0046] The specific implementation steps for this comparative example are as follows: Prepare the culture medium according to the aforementioned method. Each liter of medium contains MS dry powder, 2.0 mg of 6-benzylaminopurine, 0.8 mg of naphthaleneacetic acid, 800 mg of polyvinylpyrrolidone, 30 g of sucrose, and 7 g of carrageenan. Adjust the pH to 5.8. Do not add L-ascorbic acid. S1: Dissolve the MS dry powder, sucrose, and carrageenan in distilled water and bring the volume to 1 L. S2: Add 2.0 mg of 6-benzylaminopurine, 0.8 mg of naphthaleneacetic acid, and 800 mg of polyvinylpyrrolidone, and mix well. S3: Adjust the pH to 5.8. S4: Dispense the medium and sterilize at 121°C for 20 min. S5: After sterilization, cool to 95°C, remove the culture flasks, shake clockwise once to coat the inner wall of the flasks with the culture medium, and then lay them flat to cool and solidify.

[0047] Newly sprouted rhizomes and buds from three-year-old, disease-free *Polygonatum sibiricum* plants were selected as explants. After rinsing the rhizomes and buds thoroughly with running water, they underwent sterilization in a clean bench: first, treatment with 75% alcohol for 30 seconds, followed by rinsing once with sterile water; then, treatment with 0.1% sodium hypochlorite for 8 minutes, followed by rinsing five times with sterile water. After blotting the surface moisture with sterile filter paper, the sterilized rhizomes and bud explants were inoculated onto the aforementioned culture medium and cultured at 24±2℃, with a light intensity of 2000 lx and a photoperiod of 12h light / 12h darkness.

[0048] The results of this comparative study are as follows: After 25 days of culture, the browning rate of explants was 27.5%, and the induction rate of adventitious buds was 73.6%. Both the browning inhibition effect and the induction rate were significantly lower than those of Example 1.

[0049] A comparative analysis of the experimental results of Examples 1-3 and Comparative Examples 1-2 shows that: Regarding the browning rate of explants, the browning rate was 8.7% in Example 1, 13.9% in Example 2, and 7.6% in Example 3, while the browning rate of Comparative Example 1 was as high as 66.2%, and that of Comparative Example 2 was 27.5%. This indicates that the composite antioxidant system composed of polyvinylpyrrolidone and L-ascorbic acid added to the culture medium in this invention can effectively reduce the browning rate of explants from the rhizomes and buds of Polygonatum odoratum from northern China. Its browning inhibition effect is significantly better than that of Comparative Example 1 without added antioxidants and Comparative Example 2 with only a single antioxidant added.

[0050] Regarding the adventitious shoot induction rate, the induction rate of Example 1 was 92.3%, Example 2 was 82.3%, and Example 3 was 91.5%, while the induction rate of Comparative Example 1 was only 35.7% and that of Comparative Example 2 was 73.6%. This indicates that the synergistic ratio of 6-benzylaminopurine and naphthaleneacetic acid specified in this invention can precisely regulate the balance between cell division and cell elongation, specifically adapting to the differentiation needs of rhizome shoots of Polygonatum sibiricum in northern China, and the adventitious shoot induction rate is significantly better than that of Comparative Example 1 and Comparative Example 2.

[0051] Regarding the average number of shoots per explant, Example 1 had 5.2 shoots, Example 2 had 3.1 shoots, and Example 3 had 4.5 shoots, while Comparative Example 1 had less than 2 shoots and the shoots were thin, weak, and yellowish. This indicates that the hormone ratio of the present invention is beneficial for inducing a greater number of adventitious shoots, and the shoot quality is excellent.

[0052] Regarding the solidification stability of the culture medium, in Examples 1-3 and Comparative Example 2, carrageenan was used as the solidifying agent. After sterilization and cooling, the culture medium remained solid and stable without liquefaction. However, in Comparative Example 1, agar was used as the solidifying agent, and some of the culture medium softened during the cultivation process, increasing the risk of contamination. This indicates that replacing agar with carrageenan as the solidifying agent results in stronger solidification stability of the culture medium, which is beneficial for stable industrial seedling cultivation.

[0053] Regarding the effectiveness of the condensate treatment, Examples 1-3 and Comparative Example 2 involved sterilizing the culture flasks, cooling them to 95°C, removing them, and shaking them clockwise once. This allowed the molten culture medium to coat the inner wall of the flasks and absorb the condensate. No obvious condensate droplets were observed on the inner wall of the culture flasks, and no rotting occurred at the base of the explants. In contrast, Comparative Example 1 did not undergo this treatment, and obvious condensate droplets were visible on the inner wall of the culture flask, with some explants showing signs of rotting at their base. This indicates that the treatment process effectively eliminates condensate accumulation inside the flasks, preventing explant rotting due to soaking.

[0054] In summary, Example 1 (containing 2.5 mg of 6-benzylaminopurine, 0.8 mg of naphthaleneacetic acid, 800 mg of polyvinylpyrrolidone, 80 mg of L-ascorbic acid, 30 g of sucrose, and 7 g of carrageenan per liter of culture medium, pH 5.8) is the optimal formulation. After 15 days of culture, an induction rate of 92.3% and an average number of 5.2 buds were obtained, with low browning rate and robust buds, resulting in the best overall effect.

[0055] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A culture medium for inducing root and stem buds of Polygonatum odoratum from northern China, characterized in that, Based on MS medium, each liter of medium contains: 0.5–3.5 mg of 6-benzylaminopurine, 0.1–2.0 mg of naphthaleneacetic acid, 100–1500 mg of polyvinylpyrrolidone, 20–150 mg of L-ascorbic acid, 30 g of sucrose, and 7 g of carrageenan, and is diluted to 1 L with distilled water; the pH of the medium is 5.

8.

2. The root and stem bud induction culture medium of Polygonatum odoratum from northern China according to claim 1, characterized in that: The culture medium contains 2.5 mg of 6-benzylaminopurine, 0.8 mg of naphthaleneacetic acid, 800 mg of polyvinylpyrrolidone, and 80 mg of L-ascorbic acid per liter.

3. The rootstock bud induction culture medium of *Polygonatum sibiricum* according to claim 2, characterized in that: The culture medium was used for the primary induction culture of explants from the rhizome buds of Polygonatum odoratum from northern China.

4. A method for preparing the culture medium according to any one of claims 1 to 3, characterized in that, Includes the following steps: S1: Dissolve MS dry powder, sucrose and carrageenan in distilled water and bring the volume to 1L; S2: Add 6-benzylaminopurine, naphthaleneacetic acid, polyvinylpyrrolidone and L-ascorbic acid, and mix well; S3: Adjust pH to 5.8; S4: After dispensing, sterilize at 121℃ for 20 minutes; S5: After sterilization, cool to 95℃, remove the culture bottle, shake it clockwise once to spread the culture medium on the inner wall of the bottle, and then lay it flat to cool and solidify.

5. The method for cultivating rhizome buds of Polygonatum odoratum from northern China according to claim 4, characterized in that: The shaking time in step S5 is 3 to 5 seconds.

6. A method for cultivating rhizome buds of Polygonatum odoratum from northern China using a culture medium, characterized in that: The sterilized rhizome shoot explants were inoculated onto the culture medium described in any one of claims 1 to 3 and cultured at 24±2℃, light intensity of 2000 lx, and photoperiod of 12h light / 12h dark.

7. The method for cultivating rhizome buds of Polygonatum odoratum from northern China according to claim 6, characterized in that: The disinfection process includes treatment with 75% alcohol for 30 seconds and treatment with 0.1% sodium hypochlorite for 8 minutes.

8. A method for cultivating rhizome buds of Polygonatum odoratum from northern China according to claim 6 or 7, characterized in that: The cultivation period is 10 to 28 days.

9. A method for cultivating rhizome buds of Polygonatum odoratum from northern China according to any one of claims 1 to 3, characterized in that: The concentrations of 6-benzylaminopurine were 2.0–3.0 mg / L, naphthaleneacetic acid was 0.5–1.5 mg / L, polyvinylpyrrolidone was 500–1200 mg / L, and L-ascorbic acid was 50–120 mg / L.

10. A method for cultivating rhizome buds of Polygonatum odoratum from northern China according to claim 4 or 5, characterized in that: The shaking operation in step S5 is performed while the culture medium is in a molten state.

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