Method for rooting induction of aulaclausia serpentina tissue culture seedlings

Abstract

The application discloses a method for rooting induction of Cynanchum coccineum Schneid. tissue culture seedlings, and comprises the following steps: inoculating Cynanchum coccineum Schneid. tissue culture seedlings into MS modified culture medium to obtain sterile seedlings; inoculating the sterile seedlings into a proliferation culture medium for proliferation culture for 28-35 days to obtain adventitious bud clusters; inoculating the adventitious bud clusters into a rooting induction culture medium for rooting induction to obtain Cynanchum coccineum Schneid. seedlings; wherein the MS modified culture medium is a basic culture medium added with 0.1-0.2 g / L of activated carbon; the proliferation culture medium is a basic culture medium added with 0.2-1.0 mg / L of IBA, 0.1-0.5 mg / L of IAA and 0.3-1.0 mg / L of 6-BA; and the rooting induction culture medium is a basic culture medium added with 0.5-2.0 mg / L of NAA or 0.5-2.0 mg / L of IBA. The application can quickly induce Cynanchum coccineum Schneid. seedlings, and solves the problems of rooting difficulty and introduction and cultivation difficulty of Cynanchum coccineum Schneid. tissue culture.

Description

Technical Field

[0001] This invention belongs to the field of medicinal plant propagation technology, specifically relating to a method for inducing rooting in tissue culture seedlings of Corydalis yanhusuo. Background Technology

[0002] Corydalis ophiocarpa Hook. f. et .Thoms. is an annual herb belonging to the genus Corydalis in the Papaveraceae family. The whole plant is used medicinally. Ancient Tibetan medical books record that Corydalis ophiocarpa is also called Ye Dong Sai Guo, and it is often used as a substitute for Wa Xia Ga to treat injuries, boils, and carbuncles. Modern researchers have found that its roots can be used to treat rheumatism, and the alkaloids extracted from Corydalis ophiocarpa have antiarrhythmic, anti-myocardial ischemia, analgesic, antihypertensive, and antimalarial activities. The Chinese Materia Medica includes Corydalis ophiocarpa in prescriptions to treat blood fever, headache, chest tightness, and red eyes.

[0003] Currently, cultivating *Corydalis amabilis* through introduction into the market is quite difficult. The seed production of *Corydalis amabilis* is low, and both the seeds and plants have high requirements for their growing environment. For example, if the humidity exceeds the suitable range, mold and plant death are likely to occur. *Corydalis amabilis* has a generally low reproductive capacity, poor adaptability, slow growth rate, and is prone to death, making it challenging to expand its cultivation scale and fully develop its medicinal effects. The inventors' tissue culture experiments using *Corydalis amabilis* seeds only yielded tissue culture seedlings with stems and leaves; no root system was observed, with only a small number developing hairy roots. Long-term culture, multiple subcultures, and rooting induction failed to reveal a taproot system. Compared to related species like *Corydalis lithoides*, rooting of tissue culture seedlings is extremely difficult.

[0004] Therefore, there is an urgent need for a method to induce rooting of Corydalis rubra tissue culture seedlings that can promote the proliferation and rooting of Corydalis rubra in order to achieve its large-scale production and development. Summary of the Invention

[0005] The object of the present invention is to overcome at least the aforementioned defects and to provide advantages that will be described later.

[0006] To achieve these objectives and other advantages of the present invention, a method for inducing rooting in tissue culture seedlings of Corydalis fragilis is now provided, comprising:

[0007] Sterile seedlings of Corydalis cambogia were obtained by inoculating tissue culture seedlings into MS modified medium.

[0008] After cutting the sterile seedlings, they were inoculated into the proliferation medium and cultured for 28-35 days to obtain adventitious bud clusters.

[0009] Adventitious bud clusters were divided into individual plants and inoculated into rooting induction medium to induce rooting, thus obtaining seedlings of Corydalis rubra.

[0010] The MS modified medium uses MS solid medium as the base medium, and 0.1-0.2 g / L of activated carbon is added to the base medium by volume. The proliferation medium uses MS solid medium as the base medium, and IBA 0.2-1.0 mg / L, IAA 0.1-0.5 mg / L, and 6-BA 0.3-1.0 mg / L are added to the base medium by volume. The rooting induction medium uses MS solid medium as the base medium, and NAA 0.5-2.0 mg / L or IBA 0.5-2.0 mg / L are added to the base medium by volume.

[0011] Specifically, in the above scheme, Corydalis cambogia tissue culture seedlings are inoculated into MS solid medium supplemented with 0.1% activated carbon and cultured for 3-5 weeks, with subculture every 25 days.

[0012] The present invention relates to a method for inducing rooting of Corydalis rubra tissue culture seedlings. By aseptically cultivating Corydalis rubra tissue culture seedlings and then subjecting them to proliferation and induction cultivation, Corydalis rubra seedlings can be rapidly induced, thus solving the problem of difficulties in introducing and cultivating Corydalis rubra.

[0013] Preferably, the amount of IBA added is 0.2 mg / L, 0.5 mg / L, or 1.0 mg / L; the amount of IAA added is 0.1 mg / L, 0.2 mg / L, or 0.5 mg / L; and the amount of 6-BA added is 0.3 mg / L, 0.6 mg / L, or 1.0 mg / L. The rooting induction medium is based on MS solid medium, and based on the volume of the basal medium, 0.5 mg / L, 1.0 mg / L, or 2.0 mg / L of NAA, or 0.5 mg / L, 1.0 mg / L, or 2.0 mg / L of IBA, or IBA, or IBA, or IBA, are added to the basal medium.

[0014] In the above scheme, the method for inducing rooting of tissue culture seedlings of Corydalis fragilis in this invention achieves better proliferation effect by adjusting the proliferation medium, with a proliferation multiple of up to 8 times, laying a better foundation for expanding the planting scale of Corydalis fragilis; by adjusting the rooting medium, the diameter and length of the main root of Corydalis fragilis are induced to grow, thereby giving full play to the advantages of the main root and better absorbing nutrients, while inducing the growth of the number of lateral roots to better assist the main root in fully absorbing nutrients.

[0015] Preferably, the proliferation medium is based on MS solid medium, and IBA 0.2 mg / L, IAA 0.1 mg / L and 6-BA 0.3 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and NAA 0.5 mg / L is added to the basal medium by volume.

[0016] Preferably, the proliferation medium is based on MS solid medium, and IBA 1.0 mg / L, IAA 0.1 mg / L and 6-BA 0.6 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and NAA 0.5 mg / L is added to the basal medium by volume.

[0017] Preferably, the proliferation medium is based on MS solid medium, and IBA 1.0 mg / L, IAA 0.5 mg / L and 6-BA 1.0 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and NAA 0.5 mg / L is added to the basal medium by volume.

[0018] Preferably, the proliferation medium is based on MS solid medium, and IBA 0.2 mg / L, IAA 0.1 mg / L and 6-BA 0.3 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and IBA 1.0 mg / L is added to the basal medium by volume.

[0019] Preferably, the proliferation medium is based on MS solid medium, and IBA 1.0 mg / L, IAA 0.1 mg / L and 6-BA 0.6 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and IBA 1.0 mg / L is added to the basal medium by volume.

[0020] Preferably, the proliferation medium is based on MS solid medium, and IBA 1.0 mg / L, IAA 0.5 mg / L and 6-BA 1.0 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and IBA 1.0 mg / L is added to the basal medium by volume.

[0021] In the above-described method, the rooting induction method for tissue culture seedlings of Corydalis rubra of the Red Delicious, through adjustments to the proliferation medium and rooting induction medium, results in Corydalis rubra seedlings with distinct taproots, which facilitates taproot development and improves the transplant survival rate. Simultaneously, the acquisition of more lateral roots enhances the seedlings' resistance to adverse conditions and facilitates the absorption of nutrients from the environment, further improving the transplant survival rate.

[0022] Preferably, the conditions for proliferation culture include: during the first 1-7 days of proliferation culture, alternating between 6 hours of blue light irradiation and 5 hours of red light irradiation; during the first 8-14 days of proliferation culture, alternating between 8 hours of blue light irradiation and 6 hours of red light irradiation; and after the first 14 days of proliferation culture, alternating between 8 hours of blue light irradiation and 5 hours of red light irradiation.

[0023] In the above scheme, by adjusting the light source, the adventitious buds of Corydalis rubra are promoted, and the proliferation rate of Corydalis rubra is increased, laying a better foundation for further expanding the planting scale of Corydalis rubra.

[0024] Specifically, during the first 1-7 days of propagation culture, blue light irradiation is used to stimulate the sterile seedlings to perform photosynthesis better, thus storing more nutrients for the formation of adventitious buds. At the same time, it helps the sterile seedlings to better absorb the nutrients in the propagation culture medium and utilize the energy generated by photosynthesis to induce the formation of adventitious buds in Corydalis rubra. After the blue light irradiation is completed, red light is used to continue irradiation, giving the sterile seedlings a certain period of recovery time, while preventing the decomposition of chlorophyll during the recovery period and reducing the loss of nutrients.

[0025] During the 8-14 day propagation culture, the duration of blue light irradiation is increased to promote photosynthesis in the sterile seedlings, thereby accumulating nutrients and laying a better foundation for the production of more adventitious buds. At the same time, the duration of red light irradiation is also increased to prevent the decomposition of chlorophyll and to give the sterile seedlings sufficient time to recover.

[0026] After 14 days of propagation culture, the duration of blue light irradiation was kept constant to promote the continuous growth of adventitious buds in sterile seedlings under photosynthesis, ensure the quality of adventitious buds, and improve the survival rate of adventitious buds.

[0027] Preferably, the conditions for rooting induction include: during the first 1-5 days of rooting induction, alternating between 6 hours of strong light and 8 hours of weak light; during the first 6-20 days of rooting induction, alternating between 6 hours of ultra-strong light, 5 hours of dim light, and 5 hours of weak light; and from the 21st day onwards, alternating between 10 hours of strong light and 8 hours of dim light. The ultra-strong light concentration is 12,000-15,000 Lux; the strong light concentration is 9,000-10,000 Lux; the weak light concentration is 1-5 Lux; and the dim light concentration is below 0.5 Lux.

[0028] In the above scheme, by alternating stimulation with different light intensities, the vitality of the main root of Corydalis rubra is kept stable. While the main root grows strong, more lateral roots can be induced to grow, which can better assist the main root in fully absorbing nutrients.

[0029] Specifically, during the first 1-5 days of rooting induction, strong light is used to stimulate photosynthesis in the plants. The accumulation of photosynthetic products and the nutrient support of the rooting induction medium promote the continuous increase in the diameter of adventitious buds, which then grow into the taproot of Corydalis fragilis. Then, the strong light is switched to weak light, and the nutrients in the rooting induction medium are used to further promote the continuous growth of the taproot diameter and the continuous elongation of Corydalis fragilis to fully utilize the advantages of the taproot, thereby improving the survival rate after rooting.

[0030] During the 6-20 days of rooting induction, strong light is used to stimulate the plant to continuously accumulate nutrients through photosynthesis, and the nutrients in the rooting induction medium are used to promote the continuous growth of the diameter of the taproot. The strong light is then switched to dim light and then to weak light to stimulate the taproot to elongate and stimulate the formation of lateral roots.

[0031] 21 days after rooting induction, the generation of lateral roots in the plant is stimulated by switching between strong light and dark light, thus increasing the number of lateral roots.

[0032] Advantages of this invention:

[0033] First, the method for inducing rooting of Corydalis rubra tissue culture seedlings in this invention involves aseptic cultivation followed by proliferation and induction cultivation of Corydalis rubra tissue culture seedlings, thereby rapidly inducing Corydalis rubra seedlings and solving the problem of difficulty in introducing and cultivating Corydalis rubra.

[0034] Secondly, the method for inducing rooting of Corydalis rubra tissue culture seedlings in this invention achieves better proliferation results by adjusting the proliferation medium, with a proliferation multiple of up to 8 times, laying a better foundation for expanding the planting scale of Corydalis rubra tissue culture.

[0035] Furthermore, the method for inducing rooting in tissue culture seedlings of Corydalis rubra in this invention, by adjusting the proliferation medium and the rooting induction medium, results in Corydalis rubra seedlings with obvious taproots, which is beneficial to taproot development and improves the transplant survival rate of Corydalis rubra plants.

[0036] Furthermore, the method for inducing rooting of tissue culture seedlings of Corydalis rubra in this invention, by adjusting the proliferation medium and the rooting induction medium, results in Corydalis rubra seedlings with robust taproots and a greater number of lateral roots. This is beneficial for enhancing the seedlings' resistance to adverse conditions and for fully absorbing nutrients from the environment, thereby improving the transplant survival rate of Corydalis rubra plants.

[0037] Furthermore, the method for inducing rooting of Corydalis rubra tissue culture seedlings in this invention screens the optimal hormone concentration for inducing proliferation and rooting of Corydalis rubra tissue culture seedlings, which is of great significance for increasing the yield of Corydalis rubra seedlings, improving seedling quality, and stabilizing the medicinal properties of the herb. At the same time, it is also conducive to further improving the rapid propagation system of Corydalis rubra tissue culture and provides valuable reference for the propagation of Corydalis rubra tissue culture. Detailed Implementation

[0038] The present invention will now be described in further detail so that those skilled in the art can implement it based on the description.

[0039] Example 1

[0040] The method for inducing rooting in tissue culture seedlings of Corydalis fragilis includes the following steps:

[0041] Step 1: Using MS solid medium as the basal medium, add 0.1 g / L activated carbon to the basal medium by volume to obtain MS modified medium; using MS solid medium as the basal medium, add 0.2 mg / L IBA, 0.1 mg / L IAA, and 0.3 mg / L 6-BA to the basal medium by volume to obtain proliferation medium; using MS solid medium as the basal medium, add 0.5 mg / L NAA to the basal medium by volume to obtain rooting induction medium.

[0042] Step 2: Inoculate the tissue culture seedlings of Corydalis rubra into MS modified medium and culture for 28 days. Subculture once on day 25 to obtain sterile seedlings.

[0043] Step 3: After pruning the sterile seedlings, inoculate them into the proliferation medium and culture for 28 days to obtain adventitious bud clusters.

[0044] Step 4: Divide the adventitious bud clusters into individual plants and inoculate them into rooting induction medium to induce rooting and obtain Corydalis rubra seedlings.

[0045] The cultivation conditions were: temperature 25℃, humidity 50%, and light exposure for 12 hours.

[0046] Example 2

[0047] Based on Example 1, the proliferation medium was replaced with a medium containing 0.2 mg / L IBA + 0.2 mg / L IAA + 1.0 mg / L 6-BA.

[0048] Example 3

[0049] Based on Example 1, the proliferation medium was replaced with a medium containing 0.2 mg / L IBA + 0.5 mg / L IAA + 0.6 mg / L 6-BA.

[0050] Example 4

[0051] Based on Example 1, the proliferation medium was replaced with a medium containing 0.5 mg / L IBA + 0.1 mg / L IAA + 1.0 mg / L 6-BA.

[0052] Example 5

[0053] Based on Example 1, the proliferation medium was replaced with a medium containing 0.5 mg / L IBA + 0.2 mg / L IAA + 0.6 mg / L 6-BA.

[0054] Example 6

[0055] Based on Example 1, the proliferation medium was replaced with a medium containing 0.5 mg / L IBA + 0.5 mg / L IAA + 0.3 mg / L 6-BA.

[0056] Example 7

[0057] Based on Example 1, the proliferation medium was replaced with a medium containing 1.0 mg / L IBA + 0.1 mg / L IAA + 0.6 mg / L 6-BA.

[0058] Example 8

[0059] Based on Example 1, the proliferation medium was replaced with a medium containing 1.0 mg / L IBA + 0.2 mg / L IAA + 0.3 mg / L 6-BA.

[0060] Example 9

[0061] Based on Example 1, the proliferation medium was replaced with a medium containing 1.0 mg / L IBA + 0.5 mg / L IAA + 1.0 mg / L 6-BA.

[0062] Example 10

[0063] Based on Example 1, the rooting induction medium was replaced with a medium containing 1.0 mg / L NAA.

[0064] Example 11

[0065] Based on Example 1, the rooting induction medium was replaced with a medium containing 2.0 mg / L NAA.

[0066] Example 12

[0067] Based on Example 1, the rooting induction medium was replaced with a medium containing 0.5 mg / L IBA.

[0068] Example 13

[0069] Based on Example 1, the rooting induction medium was replaced with a medium containing 1.0 mg / L IBA.

[0070] Example 14

[0071] Based on Example 1, the rooting induction medium was replaced with a medium containing 2.0 mg / L IBA.

[0072] Example 15

[0073] The method for inducing rooting in tissue culture seedlings of Corydalis fragilis includes the following steps:

[0074] Step 1: Using MS solid medium as the basal medium, add 0.1~0.2 g / L activated carbon to the basal medium by volume to obtain MS modified medium; using MS solid medium as the basal medium, add 0.2 mg / L IBA, 0.1 mg / L IAA, and 0.3 mg / L 6-BA to the basal medium by volume to obtain proliferation medium; using MS solid medium as the basal medium, add 0.5 mg / L NAA to the basal medium by volume to obtain rooting induction medium.

[0075] Step 2: Inoculate the tissue culture seedlings of Corydalis rubra into MS modified medium and culture for 28 days. Subculture once on day 25 to obtain sterile seedlings.

[0076] Step 3: After pruning the sterile seedlings, inoculate them into the proliferation medium and culture for 28-35 days to obtain adventitious bud clusters.

[0077] Step 4: Divide the adventitious bud clusters into individual plants and inoculate them into rooting induction medium to induce rooting and obtain Corydalis rubra seedlings.

[0078] The culture environment was maintained at a temperature of 25℃ and a humidity of 50%. The conditions for proliferation culture included: during the first 1-7 days of proliferation culture, alternating between 6 hours of blue light irradiation and 5 hours of red light irradiation; during the first 8-14 days of proliferation culture, alternating between 8 hours of blue light irradiation and 6 hours of red light irradiation; and after the 14th day of proliferation culture, alternating between 8 hours of blue light irradiation and 5 hours of red light irradiation.

[0079] Example 16

[0080] The method for inducing rooting in tissue culture seedlings of Corydalis fragilis includes the following steps:

[0081] Step 1: Using MS solid medium as the basal medium, add 0.1~0.2 g / L activated carbon to the basal medium by volume to obtain MS modified medium; using MS solid medium as the basal medium, add 0.2 mg / L IBA, 0.1 mg / L IAA, and 0.3 mg / L 6-BA to the basal medium by volume to obtain proliferation medium; using MS solid medium as the basal medium, add 0.5 mg / L NAA to the basal medium by volume to obtain rooting induction medium.

[0082] Step 2: Inoculate the tissue culture seedlings of Corydalis rubra into MS modified medium and culture for 28 days. Subculture once on day 25 to obtain sterile seedlings.

[0083] Step 3: After pruning the sterile seedlings, inoculate them into the proliferation medium and culture for 28-35 days to obtain adventitious bud clusters.

[0084] Step 4: Divide the adventitious bud clusters into individual plants and inoculate them into rooting induction medium to induce rooting, thereby obtaining Corydalis rubra seedlings.

[0085] The culture environment was maintained at a temperature of 25℃ and a humidity of 50%. Rooting induction conditions included: during days 1-5 of rooting induction, alternating between 6 hours of strong light and 8 hours of weak light; during days 6-20 of rooting induction, alternating between 6 hours of extremely strong light, 5 hours of dim light, and 5 hours of weak light; and from day 21 onwards, alternating between 10 hours of strong light and 8 hours of dim light. The values ​​for extremely strong light were 12,000-15,000 Lux; strong light was 9,000-10,000 Lux; weak light was 1-5 Lux; and dim light was below 0.5 Lux.

[0086] Example 17

[0087] The method for inducing rooting in tissue culture seedlings of Corydalis fragilis includes the following steps:

[0088] Step 1: Using MS solid medium as the basal medium, add 0.1~0.2 g / L activated carbon to the basal medium by volume to obtain MS modified medium; using MS solid medium as the basal medium, add 0.2 mg / L IBA, 0.1 mg / L IAA, and 0.3 mg / L 6-BA to the basal medium by volume to obtain proliferation medium; using MS solid medium as the basal medium, add 0.5 mg / L NAA to the basal medium by volume to obtain rooting induction medium.

[0089] Step 2: Inoculate the tissue culture seedlings of Corydalis rubra into MS modified medium and culture for 28 days. Subculture once on day 25 to obtain sterile seedlings.

[0090] Step 3: After pruning the sterile seedlings, inoculate them into the proliferation medium and culture for 28-35 days to obtain adventitious bud clusters.

[0091] Step 4: Divide the adventitious bud clusters into individual plants and inoculate them into rooting induction medium to induce rooting, thereby obtaining Corydalis rubra seedlings.

[0092] The culture environment was maintained at a temperature of 25℃ and a humidity of 50%. The conditions for propagation culture included: during days 1-7, alternating between 6 hours of blue light and 5 hours of red light; during days 8-14, alternating between 8 hours of blue light and 6 hours of red light; and after day 14, alternating between 8 hours of blue light and 5 hours of red light. The conditions for rooting induction included: during days 1-5, alternating between 6 hours of strong light and 8 hours of weak light; during days 6-20, alternating between 6 hours of extremely strong light, 5 hours of dim light, and 5 hours of weak light; and from day 21 onwards, alternating between 10 hours of strong light and 8 hours of dim light. Extremely strong light was 12000-15000 Lux; strong light was 9000-10000 Lux; weak light was 1-5 Lux; and dim light was below 0.5 Lux.

[0093] Comparative Example 1

[0094] The method for inducing rooting in tissue culture seedlings of Corydalis fragilis includes the following steps:

[0095] Step 1: Inoculate the tissue culture seedlings of Corydalis cambogia into MS solid medium supplemented with 0.1% activated carbon and culture for 3-5 weeks, subculturing every 25 days to obtain sterile seedlings.

[0096] Step 2: After pruning the sterile seedlings, inoculate them into the proliferation medium and culture for 30 days to obtain adventitious bud clusters.

[0097] Step 3: Divide the adventitious bud clusters into individual plants and inoculate them into rooting induction medium to obtain Corydalis rubra seedlings.

[0098] The culture conditions were: temperature 25℃, humidity 50%, and light exposure of 12 hours. Both the proliferation medium and the rooting induction medium were MS medium.

[0099] Comparative Example 2

[0100] Based on Comparative Example 1, the proliferation medium was replaced with a medium supplemented with 0.2 mg / L IBA, 0.1 mg / L IAA, and 0.3 mg / L 6-BA.

[0101] Comparative Example 3

[0102] Based on Comparative Example 1, the proliferation medium was replaced with a medium containing 1.0 mg / L IBA.

[0103] Comparative Example 4

[0104] Based on Comparative Example 1, the proliferation medium was replaced with a medium containing 0.5 mg / L IAA.

[0105] Comparative Example 5

[0106] Based on Comparative Example 1, the proliferation medium was replaced with a medium containing 1.0 mg / L 6-BA.

[0107] Comparative experiments on root induction of Corydalis fragilis using the methods of Examples 1-17 and Comparative Examples 1-5.

[0108] The methods of Examples 1-17 and Comparative Examples 1-3 were used to conduct propagation culture and rooting induction of Corydalis rubra. The number of Corydalis rubra tissue culture seedlings in each group of propagation culture was 60. After 30 days of culture, the propagation observation was recorded, and the number of seedlings after propagation, the number of seedlings without propagation, and the number of seedlings that survived after propagation were recorded. The survival rate and propagation multiple were calculated. The results are shown in Table 1. For rooting induction, the adventitious buds of the plants obtained from propagation culture were divided into individual plants and then induced. The number of Corydalis rubra plants in each group of rooting induction was 60. After 30 days of culture, the number of dead plants, the number of rooted seedlings, and the root condition were observed and recorded. The survival rate and rooting rate after rooting were calculated. The results are shown in Table 2. Among them, the survival rate after propagation = number of seedlings that survive after propagation / number of seedlings after propagation; the propagation multiple = number of seedlings that survive after propagation / number of seedlings used for propagation; the survival rate after rooting = (number of seedlings used for rooting - number of dead seedlings) / number of seedlings used for rooting; the rooting rate = number of rooted seedlings / (number of seedlings used for rooting - number of dead seedlings).

[0109] Table 1: Proliferation Culture Status

[0110]

[0111] Table 2: Rooting Induction Results

[0112]

[0113] As shown in Table 1, the tissue culture seedlings of *Codonopsis pilosula* survived well in the proliferation medium designed in this invention. Furthermore, the proliferation rate of *Codonopsis pilosula* tissue culture seedlings after cultivation in this medium reached more than 4 times. The highest proliferation rate, exceeding 8 times, was achieved when the medium was supplemented with IBA 0.2 mg / L, IAA 0.1 mg / L, and 6-BA 0.3 mg / L. During proliferation culture, adjusting the light source effectively promoted the production of adventitious buds in *Codonopsis pilosula*, increasing the proliferation rate to 9 times.

[0114] As shown in Table 2, after the adventitious buds of *Codora sinensis* were divided into individual plants, they exhibited good rooting and survival rates in the rooting induction medium designed in this invention. Specifically, the survival rate reached 100% in the medium supplemented with 0.5 mg / L NAA and 1.0 mg / L IBA. Rooting induction in the NAA-supplemented medium increased the diameter of the taproot, giving it dominance and enhancing the seedlings' resistance to stress and nutrient absorption. However, rooting induction in a medium with IBA concentrations below 2.0 mg / L had little effect on taproot growth, resulting in thin, elongated taproots similar in diameter to the lateral roots, which was detrimental to root development. During rooting induction, adjusting the light intensity could promote taproot elongation and increase taproot diameter, while also inducing the growth of more lateral roots.

[0115] Although embodiments of the present invention have been disclosed above, they are not limited to the applications listed in the specification and embodiments. It can be applied to various fields suitable for the present invention. Other modifications can be readily implemented by those skilled in the art. Therefore, the present invention is not limited to the specific details without departing from the general concept defined by the claims and their equivalents.

Claims

1. A method for inducing rooting in tissue culture seedlings of Corydalis rubra, characterized in that, include: Sterile seedlings of Corydalis cambogia tissue culture were obtained by inoculating them into MS modified medium. After cutting the sterile seedlings, they were inoculated into the proliferation medium and cultured for 28-35 days to obtain adventitious bud clusters. Adventitious bud clusters were divided into individual plants and inoculated into rooting induction medium for rooting induction to obtain Corydalis rubra seedlings; The MS modified medium uses MS solid medium as the base medium, and 0.1~0.2 g / L of activated carbon is added to the base medium based on the volume of the base medium. The proliferation medium is based on MS solid medium, and IBA 0.2~1.0 mg / L, IAA 0.1~0.5 mg / L, and 6-BA 0.3~1.0 mg / L are added to the basic medium by volume. The rooting induction medium is based on MS solid medium, and NAA 0.5~2.0 mg / L or IBA 0.5~2.0 mg / L is added to the basic medium by volume. The conditions for proliferation culture include: During the first 1-7 days of proliferation culture: alternate between 6 hours of blue light irradiation and 5 hours of red light irradiation; During the 8-14 day propagation culture: blue light irradiation for 8 hours and red light irradiation for 6 hours were used alternately. After 14 days of proliferation culture: blue light irradiation for 8 hours and red light irradiation for 5 hours were applied alternately. The conditions for rooting induction include: During the first 1-5 days of rooting induction: alternate between strong light exposure for 6 hours and weak light exposure for 8 hours. During the 6-16 days of rooting induction: alternate between 6 hours of intense light, 5 hours of dim light, and 5 hours of weak light. During the 17-20 day period of rooting induction: alternate between 6 hours of intense light irradiation and 8 hours of weak light irradiation. Rooting induction 21 days and beyond: alternate between 10 hours of strong light irradiation and 8 hours of dim light irradiation; Among them, super strong light is 12,000~15,000 Lux; strong light is 9,000~10,000 Lux; weak light is 1~5 Lux; and dark light is less than 0.5 Lux.

2. The method for inducing rooting of tissue culture seedlings of Corydalis rubra as described in claim 1, characterized in that, The proliferation medium was based on MS solid medium. Based on the volume of the basal medium, the addition amount of IBA was 0.2 mg / L, 0.5 mg / L, or 1.0 mg / L; the addition amount of IAA was 0.1 mg / L, 0.2 mg / L, or 0.5 mg / L; and the addition amount of 6-BA was 0.3 mg / L, 0.6 mg / L, or 1.0 mg / L. The rooting induction medium is based on MS solid medium. Based on the volume of the basal medium, NAA 0.5 mg / L, NAA 1.0 mg / L, NAA 2.0 mg / L, IBA 0.5 mg / L, IBA 1.0 mg / L, or IBA 2.0 mg / L are added to the basal medium.

3. The method for inducing rooting of Corydalis rubra tissue culture seedlings as described in claim 1, characterized in that, The proliferation medium is based on MS solid medium, and IBA 0.2 mg / L, IAA 0.1 mg / L and 6-BA 0.3 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and NAA 0.5 mg / L is added to the basal medium by volume.

4. The method for inducing rooting of Corydalis rubra tissue culture seedlings as described in claim 1, characterized in that, The proliferation medium is based on MS solid medium, and IBA 1.0 mg / L, IAA 0.1 mg / L and 6-BA 0.6 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and NAA 0.5 mg / L is added to the basal medium by volume.

5. The method for inducing rooting of Corydalis rubra tissue culture seedlings as described in claim 1, characterized in that, The proliferation medium was based on MS solid medium, and IBA 1.0 mg / L, IAA 0.5 mg / L and 6-BA 1.0 mg / L were added to the basic medium by volume. The rooting induction medium was based on MS solid medium, with NAA 0.5 mg / L added to the basal medium by volume.

6. The method for inducing rooting of Corydalis rubra tissue culture seedlings as described in claim 1, characterized in that, The proliferation medium is based on MS solid medium, and IBA 0.2 mg / L, IAA 0.1 mg / L and 6-BA 0.3 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and IBA 1.0 mg / L is added to the basal medium by volume.

7. The method for inducing rooting of Corydalis rubra tissue culture seedlings as described in claim 1, characterized in that, The proliferation medium is based on MS solid medium, and IBA 1.0 mg / L, IAA 0.1 mg / L and 6-BA 0.6 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and IBA 1.0 mg / L is added to the basal medium by volume.

8. The method for inducing rooting of Corydalis rubra tissue culture seedlings as described in claim 1, characterized in that, The proliferation medium is based on MS solid medium, and IBA 1.0 mg / L, IAA 0.5 mg / L and 6-BA 1.0 mg / L are added to the basal medium by volume; the rooting induction medium is based on MS solid medium, and IBA 1.0 mg / L is added to the basal medium by volume.

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