Tissue culture and rapid propagation method of corydalis nepalensis

Abstract

The invention discloses a tissue culture and rapid propagation method of corydalis nepalensis, and relates to the technical field of plant culture, according to the method, a leaf of the corydalis nepalensis is used as an explant, a tissue culture seedling of the corydalis nepalensis can be obtained only through two steps of induced differentiation culture and seedling strengthening and rooting culture, and compared with conventional tissue culture, one process is omitted. By means of the method, a large number of tissue culture seedlings of the corydalis nepalensis can be provided in a short time, compared with the prior art, the method has the advantages that materials are convenient to take, cost is saved, and the period is shortened, and a foundation is laid for overcoming artificial breeding barriers of the corydalis nepalensis and achieving industrial production of the seedlings.

Description

Technical Field

[0001] The present invention relates to the technical field of plant cultivation, and particularly relates to a method for tissue culture and rapid propagation of Corydalis hendersonii Hemsl. Background Art

[0002] Corydalis hendersonii Hemsl is a perennial herbaceous plant of the genus Corydalis in the Papaveraceae family. It is mainly distributed in the area with an altitude of 4200 - 5200 m in Tibet, China. It is the main origin of the Tibetan medicine Corydalis acuminata Franch. It has the effects of clearing heat and inflammation, cooling blood and relieving diarrhea, and can be used to treat high altitude polycythemia, ulcer pain, vasculitis, enteritis and other diseases.

[0003] At present, in addition to nearly a hundred hospital preparation products of Tibetan medicine medical institutions within Tibet, more than 40 kinds of Tibetan medicine products with national drug approval numbers, such as Honghua Ruyi Pills, Shiwei Longdan Flower Granules, Zhituo Jiebai Pills, Shisanwei Qinglan Powder, Ershiwuwei Guijiu, and Shiwei Longdan Flower Granules, etc., need to use Corydalis hendersonii Hemsl. The annual market demand exceeds 10 tons. Due to the extremely low natural reproduction ability and long growth cycle of Corydalis hendersonii Hemsl, coupled with the restriction of the extreme plateau environment, long-term predatory excavation has caused serious damage to its wild population, and the resources are facing the risk of exhaustion. Currently, it has been listed as a first-class endangered Tibetan medicine resource. At present, Corydalis hendersonii Hemsl still supplies the market with wild resources, and the supply is seriously insufficient. Some enterprises are facing the dilemma of having no medicine available.

[0004] Chinese Patent with publication number CN108522282A and publication date September 14, 2018 discloses a tissue culture method for the endangered Tibetan medicine Corydalis acuminata Franch. The steps include: (1) Selecting robust plant buds, first quickly rinsing them with ethanol, and then disinfecting them with HgC12 solution; (2) Rinsing the buds processed in step (1) with sterile water, and then sucking dry the water on the surface of the buds to obtain sterile explants for later use; (3) Under sterile conditions, inoculating the sterile explants described in step (2) onto the culture medium, first performing dark culture, and then transferring them to the cluster bud culture medium, and culturing and inducing cluster buds under light conditions to form cluster buds; (4) Transferring the cluster buds into the rooting culture medium, and obtaining rooted test-tube seedlings under light conditions; (5) Acclimatizing and transplanting the rooted test-tube seedlings into the substrate, culturing them into plants, and then planting them in the wild until harvest. However, the tissue culture process of this method requires the use of highly toxic reagents (mercuric chloride), and the explants used are buds (Corydalis hendersonii Hemsl are all underground buds, which are difficult to obtain, limited in quantity, and sampling causes great damage to the mother plants, even leading to death). The entire cultivation process needs to go through three key cultivation stages: dark culture, bud induction culture, and rooting culture. The process is cumbersome, the cycle is long, and the labor cost is high. Summary of the Invention

[0005] The object of the present invention is to provide a tissue culture and rapid propagation method for Corydalis hendersonii, using leaves as the material, which is not only convenient for material collection and will not cause irreversible damage to the mother plant, so this application has great advantages in terms of material collection; the sterilizing agent used in the present invention is sodium hypochlorite, which belongs to daily disinfection supplies, is safe and environmentally friendly, has no potential safety hazards during the operation process, and at the same time, this reagent is easy to purchase and has a low cost. This application only requires two steps of induction and differentiation culture and strong seedling rooting culture, that is, the callus induction and adventitious bud differentiation processes are completed simultaneously on the induction and differentiation medium, omitting the transfer process, which can not only greatly reduce the consumption of the medium, but also significantly reduce the labor cost and shorten the culture cycle.

[0006] The present invention is achieved through the following technical solutions:

[0007] A tissue culture and rapid propagation method for Corydalis hendersonii, comprising the following steps:

[0008] Step 1, explant acquisition: Take the young and tender leaves of Corydalis hendersonii, disinfect them and use them as explants for standby;

[0009] Step 2, induction and differentiation: In a clean bench, use a scalpel to transversely cut the above explants at an interval of 0.5 - 0.8 cm, and then inoculate the cut leaves into the induction and differentiation medium for culturing for 30 - 45 days to obtain cluster buds;

[0010] Step 3, strong seedling rooting: In a clean bench, divide the cluster buds obtained in Step 2 into small bud clusters with 3 - 5 leaves, and transfer them to the strong seedling rooting medium for culturing for 30 days to obtain rooted tissue culture seedlings;

[0011] Step 4, seedling hardening: Take out the rooted tissue culture seedlings from the culture bottle, wash the root medium, then transplant them into the seedling hardening substrate, cover the upper part with a transparent moisture-keeping cover, remove the moisture-keeping cover on the 15th day of seedling hardening, spray a nutrient solution once every 5 days during the seedling hardening period, and after culturing for 30 days, they can be transplanted into the field.

[0012] Preferably, the disinfection method in Step 1 is to sterilize the young and tender leaves of Corydalis hendersonii with alcohol for 30 seconds first, then rinse them with sterile water for 1 - 2 times, then sterilize them with a sodium hypochlorite solution for 5 minutes, and finally rinse them with sterile water for 3 - 5 times.

[0013] Preferably, the concentration of the alcohol is 75%, and the concentration of the sodium hypochlorite solution is 2 - 3%.

[0014] Preferably, in Step 2, the composition of the induction and differentiation medium includes: 0.5 - 2.0 mg / L of 6 - BA, 0.5 - 1.0 mg / L of NAA, 0.1 - 1.0 mg / L of IBA, 30 g / L of sucrose, 7 g / L of agar, the balance is MS, and the pH is 5.8 - 6.0.

[0015] Preferably, in the second step, the culture conditions for induction and differentiation are as follows: temperature is 20-24°C, light intensity is 1900-2100 lx, and light duration is 12 h / d.

[0016] Preferably, in the third step, the composition of the strong seedling rooting medium includes: 0.01-0.5 mg / L of IBA, 0.01-0.5 mg / L of NAA, 20 g / L of sucrose, 7 g / L of agar, the balance being 1 / 2 MS, and the pH is 5.8-6.0.

[0017] Preferably, in the third step, the culture conditions for strong seedling rooting are as follows: temperature is 20-24°C, light intensity is 1900-2100 lx, and light duration is 12 h / d.

[0018] Preferably, in the fourth step, the nutrient solution is 1 / 10 MS solution.

[0019] Preferably, in the fourth step, the temperature is controlled at 15-20°C under natural light.

[0020] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0021] 1. A tissue culture and rapid propagation method of Corydalis hendersonii provided in the present invention uses leaves as materials, which are not only convenient to obtain materials, but also will not cause irreversible damage to the mother plants. Therefore, this application has great advantages in terms of material collection; the sterilizing agent used in the present invention is sodium hypochlorite, which belongs to daily disinfection supplies, is safe and environmentally friendly, has no safety hazards during the operation process, and at the same time, this reagent is easy to purchase and has a low cost. This application only requires two steps of induction and differentiation culture and strong seedling rooting culture, that is, the callus induction and adventitious bud differentiation processes are completed simultaneously on the induction and differentiation medium, omitting the transfer process, which can not only greatly reduce the consumption of the medium, but also significantly reduce the labor cost and shorten the culture cycle. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 is a schematic diagram of the inoculation of the explant of Corydalis hendersonii in the invention;

[0023] Figure 2 is a schematic diagram of the cluster buds of Corydalis hendersonii (25 days after induction) in the invention;

[0024] Figure 3 is a schematic diagram of the cluster buds of Corydalis hendersonii (35 days after induction) in the invention;

[0025] Figure 4 is a schematic diagram of the proliferation of the cluster buds of Corydalis hendersonii in the invention;

[0026] Figure 5 is a schematic diagram of the strong seedlings and rooting of Corydalis hendersonii in the invention;

[0027] Figure 6 Schematic diagram of seedling acclimation of *Corydalis hendersonii* in the invention. Specific embodiments

[0028] The present invention will be further described in detail below in conjunction with embodiments, but the embodiments of the present invention are not limited thereto.

[0029] In the following embodiments, MS is the MS medium prepared with purchased MS powder, and the content of MS powder in the medium is 4.74 g / L; the 1 / 2MS used refers to the medium in which the concentration of all components of the MS medium is halved (that is, the content of MS powder is 2.37 g / L); the 1 / 10MS used refers to the medium in which the concentration of all components of the MS medium is reduced by 10 times (that is, the content of MS powder is 0.47 g / L). 6-BA refers to 6-benzylaminopurine; IBA refers to indolebutyric acid; NAA is naphthaleneacetic acid. MS, 6-BA, IBA, NAA, sucrose, and agar are all purchased from Shenzhen Zhongke Qianjin Technology Co., Ltd.

[0030] Example 1

[0031] As Figures 1 - 6 shown, a tissue culture and rapid propagation method for *Corydalis hendersonii* includes the following steps:

[0032] Step 1, explant acquisition: Take the young and tender leaves of *Corydalis hendersonii*, disinfect them and use them as explants for standby; the disinfection method is to first sterilize the young and tender leaves of *Corydalis hendersonii* with alcohol for 30 seconds, then rinse them once with sterile water, then sterilize them with sodium hypochlorite solution for 5 minutes, and finally rinse them 3 times with sterile water; the concentration of the alcohol is 75%, and the concentration of the sodium hypochlorite solution is 2%;

[0033] Step 2, induction and differentiation: In a clean bench, cut the above explants into small pieces at an interval of 0.5 cm with a scalpel, and then inoculate the cut small pieces into the induction and differentiation medium for culturing for 30 days; the composition of the induction and differentiation medium includes: 2.0 mg / L of 6-BA, 1.0 mg / L of NAA, 1.0 mg / L of IBA, 30 g / L of sucrose, 7 g / L of agar, and the balance is MS, and the pH is 5.8; the culture conditions for induction and differentiation are: the temperature is 24 °C, the light intensity is 2100 lx, and the light time is 12 h / d;

[0034] Step 3. Strengthening seedlings and promoting rooting: In a clean bench, divide the clustered buds obtained in Step 2 into small bud clusters with 3 leaves, and transfer them to the seedling strengthening and rooting medium for culturing for 30 days; the composition of the seedling strengthening and rooting medium includes: 0.5 mg / L of IBA, 0.5 mg / L of NAA, 20 g / L of sucrose, 7 g / L of agar, the balance is 1 / 2 MS, and the pH is 5.8; the culture conditions for strengthening seedlings and promoting rooting are: the temperature is 24 °C, the light intensity is 2100 lx, and the light time is 12 h / day;

[0035] Step 4. Hardening off: Take out the rooted tissue culture seedlings from the culture bottle, wash the root medium, then transplant them into the hardening off substrate, cover the upper part with a transparent plastic cup for moisturizing, then place the seedling tray in the greenhouse, control the temperature at 20 °C, and the light is natural light; Spray 1 / 10 MS nutrient solution once every 5 days during the hardening off period; On the 15th day of hardening off, remove the plastic cup above the seedling tray, and keep watering once in the morning and once in the evening, and carry out field transplantation after 30 days of hardening off.

[0036] Example 2

[0037] A tissue culture and rapid propagation method for Corydalis hendersonii Hemsl. includes the following steps:

[0038] Step 1. Obtaining explants: Take the young leaves of Corydalis hendersonii Hemsl., disinfect them and use them as explants for standby; the disinfection method is to sterilize the young leaves of Corydalis hendersonii Hemsl. with alcohol for 30 seconds first, then rinse them with sterile water 2 times, then sterilize them with sodium hypochlorite solution for 5 minutes, and finally rinse them with sterile water 4 times; the concentration of the alcohol is 75%, and the concentration of the sodium hypochlorite solution is 3%;

[0039] Step 2. Inducing differentiation: In a clean bench, cut the above explants into small pieces at an interval of 0.6 cm with a scalpel, and then inoculate the cut small pieces into the induction and differentiation medium for culturing for 38 days; the composition of the induction and differentiation medium includes: 1.25 mg / L of 6-BA, 0.75 mg / L of NAA, 0.5 mg / L of IBA, 30 g / L of sucrose, 7 g / L of agar, the balance is MS, and the pH is 5.9; the culture conditions for inducing differentiation are: the temperature is 22 °C, the light intensity is 2000 lx, and the light time is 12 h / day;

[0040] Step 3. Strengthening seedlings and promoting rooting: In a clean bench, divide the clustered buds obtained in Step 2 into small bud clusters with 5 leaves, and transfer them to the seedling strengthening and rooting medium for culturing for 30 days; the composition of the seedling strengthening and rooting medium includes: 0.2 mg / L of IBA, 0.2 mg / L of NAA, 20 g / L of sucrose, 7 g / L of agar, the balance is 1 / 2 MS, and the pH is 5.9; the culture conditions for strengthening seedlings and promoting rooting are: the temperature is 22 °C, the light intensity is 2000 lx, and the light time is 12 h / day;

[0041] Step 4. Hardening-off: Take out the rooted tissue culture seedlings from the culture bottle, wash the root medium, then transplant them into the hardening-off substrate, cover with a plastic film arch shed above for moisture preservation, then place the seedling tray in the greenhouse, control the temperature at 18°C, and the light is natural light; During the hardening-off period, spray 1 / 10 MS nutrient solution once every 5 days; On the 15th day of hardening-off, remove the plastic film above the seedling tray, and keep watering once in the morning and once in the evening, and carry out field transplantation after 30 days of hardening-off.

[0042] Example 3

[0043] A tissue culture and rapid propagation method for Corydalis hendersonii Hemsl. var. hendersonii, comprising the following steps:

[0044] Step 1. Explant acquisition: Take the young and tender leaves of Corydalis hendersonii Hemsl. var. hendersonii, disinfect them and use them as explants for standby; The disinfection method is to first sterilize the young and tender leaves of Corydalis hendersonii Hemsl. var. hendersonii with alcohol for 30 seconds, then rinse with sterile water 2 times, then sterilize with sodium hypochlorite solution for 5 minutes, and finally rinse with sterile water 5 times; The concentration of the alcohol is 75%, and the concentration of the sodium hypochlorite solution is 2%;

[0045] Step 2. Induction and differentiation: In a clean workbench, cut the above explants into small pieces at intervals of 0.8 cm with a scalpel, and then inoculate the cut small pieces into the induction and differentiation medium for cultivation for 45 days; The composition of the induction and differentiation medium includes: 0.5 mg / L of 6-BA, 0.5 mg / L of NAA, 0.1 mg / L of IBA, 30 g / L of sucrose, 7 g / L of agar, the balance is MS, and the pH is 6.0; The culture conditions for induction and differentiation are: the temperature is 20°C, the light intensity is 1900 lx, and the light time is 12 h / day;

[0046] Step 3. Seedling strengthening and rooting: In a clean workbench, divide the clustered buds obtained in Step 2 into small bud clusters with 3 - 5 leaves, and transfer them to the seedling strengthening and rooting medium for cultivation for 30 days; The composition of the seedling strengthening and rooting medium includes: 0.01 mg / L of IBA, 0.01 mg / L of NAA, 20 g / L of sucrose, 7 g / L of agar, the balance is 1 / 2 MS, and the pH is 6.0; The culture conditions for seedling strengthening and rooting are: the temperature is 20°C, the light intensity is 1900 - lx, and the light time is 12 h / day;

[0047] Step 4. Hardening-off: Take out the rooted tissue culture seedlings from the culture bottle, wash the root medium, then transplant them into the hardening-off substrate, cover with a transparent plastic cover above for moisture preservation, then place the seedling tray in the greenhouse, control the temperature at 18°C, and the light is natural light; During the hardening-off period, spray 1 / 10 MS nutrient solution once every 5 days; On the 15th day of hardening-off, remove the plastic cover, and keep watering once in the morning and once in the evening, and carry out field transplantation after 30 days of hardening-off.

[0048] Control Example 1

[0049] The difference between this comparative example and Example 1 lies in that the composition of the induction and differentiation medium includes: 2.0 mg / L of 6-BA, 1.0 mg / L of NAA, 30 g / L of sucrose, 7 g / L of agar, with the balance being MS, and the pH being 5.8.

[0050] Comparative Example 2

[0051] The difference between this comparative example and Example 1 lies in that the composition of the induction and differentiation medium includes: 2.0 mg / L of 6-BA, 1.5 mg / L of NAA, 1.0 mg / L of IBA, 30 g / L of sucrose, 7 g / L of agar, with the balance being MS, and the pH being 5.8.

[0052] Comparative Example 3

[0053] The difference between this comparative example and Example 1 lies in that the culture conditions for induction and differentiation are: temperature 26°C, light intensity 2200 lx, and light duration 12 h / d.

[0054] Comparative Example 4

[0055] The difference between this comparative example and Example 1 lies in that the culture conditions for induction and differentiation are: temperature 18°C, light intensity 2100 lx, and light duration 12 h / d.

[0056] Comparative Example 5

[0057] The difference between this comparative example and Example 1 lies in that the composition of the rooting medium includes: 0.6 mg / L of IBA, 0.6 mg / L of NAA, 20 g / L of sucrose, 7 g / L of agar, with the balance being 1 / 2 MS, and the pH being 5.8.

[0058] Comparative Example 6

[0059] The difference between this comparative example and Example 1 lies in that the composition of the rooting medium includes: 0.005 mg / L of IBA, 0.005 mg / L of NAA, 20 g / L of sucrose, 7 g / L of agar, with the balance being 1 / 2 MS, and the pH being 5.8.

[0060] Comparative Example 7

[0061] The difference between this comparative example and Example 1 lies in that the culture conditions for strong seedling rooting are: temperature 26°C, light intensity 2200 lx, and light duration 12 h / d.

[0062] Comparative Example 8

[0063] The difference between this comparative example and Example 1 lies in that the culture conditions for strong seedling rooting are: temperature 18°C, light intensity 2100 lx, and light duration 12 h / d.

[0064] Comparative Example 9

[0065] The difference between this comparative example and Example 1 is that the temperature is controlled at 25°C.

[0066] Comparative Example 10

[0067] The difference between this comparative example and Example 1 is that the temperature is controlled at 12°C.

[0068] Comparative Example 11

[0069] The difference between this comparative example and Example 1 is that during the seedling hardening period, 1 / 10 MS nutrient solution is sprayed once every 3 days.

[0070] Comparative Example 12

[0071] The difference between this comparative example and Example 1 is that during the seedling hardening period, 1 / 10 MS nutrient solution is sprayed once every 7 days.

[0072] Comparative Example 13

[0073] The difference between this comparative example and Example 1 is that during the seedling hardening period, 1 / 20 MS nutrient solution is sprayed once every 5 days.

[0074] Comparative Example 14

[0075] The difference between this comparative example and Example 1 is that during the seedling hardening period, 1 / 5 MS nutrient solution is sprayed once every 5 days.

[0076] The following table is a data table of the induction rate, rooting rate, and seedling hardening survival rate in Examples 1 - 3 and Comparative Examples 1 - 14.

[0077] Induction rate (%) Rooting rate (%) Survival rate of acclimatized seedlings (%) Example 1 81.5 88.3 91.5 Example 2 85.3 82.6 89.7 Example 3 78.5 90.6 88.7 Comparative Example 1 70.3 85.3 88.7 Comparative Example 2 67.7 86.8 90.8 Comparative Example 3 60.3 79.4 87.7 Comparative Example 4 70.9 84.5 92.6 Comparative Example 5 81.5 43.7 76.1 Comparative Example 6 81.5 11.6 66.8 Comparative Example 7 81.5 68.9 87.9 Comparative Example 8 81.5 71.6 90.2 Comparative Example 9 81.5 88.3 78.6 Comparative Example 10 81.5 88.3 82.8 Comparative Example 11 81.5 88.3 78.9 Comparative Example 12 81.5 88.3 85.3 Comparative Example 13 81.5 88.3 88.6 Comparative Example 14 81.5 88.3 78.5

[0078] Compared with Example 1, in Comparative Example 1, IBA in the induction differentiation medium plays an important role during the induction differentiation process; compared with Example 2, in Comparative Example 2, the concentration of NAA in the induction differentiation medium needs to be within a specific range during the induction differentiation process for better induction differentiation effect; compared with Example 1, in Comparative Examples 3 and 4, the induction differentiation temperature and light intensity play important roles during the induction differentiation process; compared with Example 1, in Comparative Examples 5 and 6, IBA and NAA in the strong seedling rooting medium during the rooting culture need to be within a specific range for better rooting culture effect; compared with Example 1, in Comparative Examples 7 and 8, the rooting culture temperature and light intensity play important roles during the rooting culture process; compared with Example 1, in Comparative Examples 9 and 10, the seedling hardening temperature plays an important role during the seedling hardening process; compared with Example 1, in Comparative Examples 11 - 14, the spraying interval and the amount of nutrient solution during the spraying of the nutrient solution during the seedling hardening period need to be within a specific range for a higher seedling hardening survival rate.

[0079] The above are only the preferred embodiments of the present invention, and do not impose any formal restrictions on the present invention. Any simple modifications or equivalent changes made to the above embodiments based on the technical essence of the present invention all fall within the protection scope of the present invention.

Claims

1. A tissue culture and rapid propagation method of Corydalis hendersonii Hemsl., characterized in that, It includes the following steps: Step 1, explant acquisition: Take the young and tender leaves of Corydalis hendersonii, disinfect them, and reserve them as explants; Step 2, induction and differentiation: In a clean bench, use a scalpel to transversely cut the above explants at an interval of 0.5 - 0.8 cm, and then inoculate the cut leaves into the induction and differentiation medium for 30 - 45 days to obtain cluster buds; Step 3, seedling strengthening and rooting: In a clean bench, divide the cluster buds obtained in Step 2 into small bud clusters with 3 - 5 leaves, and transfer them to the seedling strengthening and rooting medium for 30 days to obtain rooted tissue culture seedlings; Step 4, acclimatization: Take out the rooted tissue culture seedlings from the culture bottle, wash the root medium, then transplant them into the acclimatization substrate, cover it with a transparent moisture - retaining cover above. Remove the moisture - retaining cover on the 15th day of acclimatization. Spray the nutrient solution once every 5 days during the acclimatization period. After 30 days of cultivation, it can be transplanted to the field.

2. The tissue culture and rapid propagation method of Corydalis hendersonii Hemsl. according to claim 1, wherein: The disinfection method in Step 1 is to sterilize the young and tender leaves of Corydalis hendersonii with alcohol for 30 seconds first, then rinse them with sterile water 1 - 2 times, then sterilize them with sodium hypochlorite solution for 5 minutes, and finally rinse them with sterile water 3 - 5 times.

3. The tissue culture and rapid propagation method of Corydalis hendersonii Hemsl. according to claim 2, characterized in that: The concentration of the alcohol is 75%, and the concentration of the sodium hypochlorite solution is 2 - 3%.

4. The tissue culture and rapid propagation method of Corydalis hendersonii Hemsl. according to claim 1, characterized in that: In Step 2, the composition of the induction and differentiation medium includes: 0.5 - 2.0 mg / L of 6 - BA, 0.5 - 1.0 mg / L of NAA, 0.1 - 1.0 mg / L of IBA, 30 g / L of sucrose, 7 g / L of agar, the balance is MS, and the pH is 5.8 - 6.

0.

5. A tissue culture and rapid propagation method of Corydalis hendersonii Hemsl., according to claim 4, characterized in that: In Step 2, the culture conditions for induction and differentiation are: the temperature is 20 - 24°C, the light intensity is 1900 - 2100 lx, and the light time is 12 h / d.

6. The tissue culture and rapid propagation method of Corydalis hendersonii Hemsl. according to claim 1, characterized in that: In Step 3, the composition of the seedling strengthening and rooting medium includes: 0.01 - 0.5 mg / L of IBA, 0.01 - 0.5 mg / L of NAA, 20 g / L of sucrose, 7 g / L of agar, the balance is 1 / 2MS, and the pH is 5.8 - 6.

0.

7. A tissue culture and rapid propagation method of Corydalis hendersonii Hemsl., according to claim 1, characterized in that: In Step 3, the culture conditions for seedling strengthening and rooting are: the temperature is 20 - 242°C (there may be a mistake here, assuming it should be 20 - 24°C), the light intensity is 1900 - 2100 lx, and the light time is 12 h / d.

8. The tissue culture and rapid propagation method of Corydalis hendersonii Hemsl. according to claim 1, characterized in that: In Step 4, the nutrient solution is 1 / 10MS solution.

9. A tissue culture and rapid propagation method of Corydalis hendersonii Hemsl., according to claim 8, characterized in that: In Step 4, the temperature is controlled at 15 - 20°C, with natural light.

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