A rapid propagation method for Sedum sarmentosum via tissue culture with minerals

Abstract

This invention provides a method for rapid propagation of *Sedum aizoon* via tissue culture, belonging to the field of plant tissue culture technology. The method includes the following steps: 1) Using *Sedum aizoon* stem segments as explants, the stem segments are disinfected, cut, and inoculated into a callus induction medium for callus induction culture to obtain callus tissue; 2) The obtained callus tissue is inoculated into a shoot induction medium for shoot induction culture to obtain adventitious shoots; 3) The obtained adventitious shoots are inoculated into a shoot proliferation medium for shoot proliferation culture to obtain regenerated seedlings; 4) The regenerated seedlings are inoculated into a rooting medium for root induction culture to obtain transplanted seedlings; 5) The transplanted seedlings are transplanted into soil for cultivation. The method provided by this invention can significantly improve the differentiation rate of callus tissue and the survival rate of regenerated seedlings.

Description

Technical Field

[0001] This invention belongs to the field of plant tissue culture technology, and in particular relates to a method for rapid propagation of Sedum sarmentosum via tissue culture. Background Technology

[0002] Sedum sarcodactylon Sedum plumbizincicola It is a new species of Sedum in the Crassulaceae family, a perennial succulent herb that mainly reproduces asexually; it has wide adaptability, large biomass, and fast growth rate. Sedum sarmentosum can hyperaccumulate zinc and cadmium, and has great application potential.

[0003] Seed propagation and cutting propagation are the main traditional methods of propagating Sedum sarmentosum. However, these methods often result in the separation of desirable traits, difficulty in preserving the inherent characteristics of the parent plants, and the occurrence of variations. In addition, the propagation coefficient is low and the growth rate is relatively slow. Large-scale seedling cultivation and shortening the seedling cycle are urgent problems that need to be solved for the promotion and application of Sedum sarmentosum.

[0004] For rapid tissue culture propagation methods of *Sedum aizoon*, existing techniques mostly use leaves as explants. For example, the paper "Induction of Callus and Plant Regeneration from *Sedum aizoon* Leaves" (doi:10.11833 / j.issn.2095-0756.2018.03.024) describes the use of young leaves of *Sedum aizoon* as explants to induce callus and plant regeneration. This experiment used 300 mg·L⁻¹ of [unspecified ingredient]. －1 MS medium containing hydrolyzed casein was used as the basal medium. The medium was first sterilized with 75.0% ethanol for 30 seconds, followed by rinsing with sterile water 1-2 times. Then, it was sterilized with 1.5% sodium hypochlorite for 10 minutes, followed by rinsing with sterile water 2-3 times. After drying on sterile filter paper, the explants were treated. Different concentrations of 2,4-dichlorophenoxyacetic acid and 6-benzyladenine were used to induce callus formation, callus differentiation, shoot proliferation, and root induction in young *Sedum aizoon* leaves. Finally, hardening and transplanting were performed. Using young *Sedum aizoon* leaves as explants required strict selection of materials. Over-sterilization of young *Sedum aizoon* leaves during sterile treatment could easily lead to leaf inactivation, while short sterilization times could not completely eliminate bacteria on the leaves, making sterile treatment challenging. Although the best callus induction rate of this technology was 92.01%, the callus differentiation rate was only 27.44%, indicating that the vast majority of Sedum sarmentosum callus could not differentiate into adventitious buds. This also resulted in the waste of most of the young leaves of Sedum sarmentosum and the callus induced from them. Summary of the Invention

[0005] In view of this, the purpose of this invention is to provide a rapid propagation method for Sedum sarmentosum tissue culture with high callus differentiation rate.

[0006] This invention provides a method for rapid propagation of Sedum sarmentosum via tissue culture, comprising the following steps:

[0007] 1) Using the stem segments of Sedum sarmentosum as explants, the stem segments were disinfected, cut, and then inoculated into callus induction culture medium to induce callus culture and obtain callus tissue.

[0008] 2) The obtained callus tissue was inoculated into a bud induction medium for bud induction culture to obtain adventitious buds;

[0009] 3) The obtained adventitious buds are inoculated into bud proliferation medium for bud proliferation culture to obtain regenerated seedlings;

[0010] 4) The regenerated seedlings are inoculated into a rooting medium for rooting induction culture to obtain transplanted seedlings;

[0011] 5) Transplant the seedlings into nutrient soil for cultivation;

[0012] The callus induction culture medium was MS + 0.45~0.55 mg / L NAA + 2.8~3.2 mg / L 6-BA;

[0013] The bud induction medium was MS + 0.15~0.25 mg / L NAA + 2.8~3.2 mg / L 6-BA;

[0014] The bud proliferation medium was MS + 0.15~0.25 mg / L NAA + 2.8~3.2 mg / L 6-BA;

[0015] The rooting medium was MS + 0.95~1.05 mg / L IBA.

[0016] Preferably, the stem segment disinfection involves immersing the stem segments in 70-80% alcohol for 25-35 seconds, rinsing 4-5 times, then placing them in 0.09-0.11 wt% HgCl2 solution for 6-8 minutes, and rinsing with water 4-5 times.

[0017] Preferably, the stem segment disinfection process includes a cleaning step, in which gauze soaked in laundry detergent is used for cleaning, followed by rinsing with running water for 25-35 minutes.

[0018] Preferably, the length of the cut stem segment is 0.45~0.55cm.

[0019] Preferably, the temperature for inducing and culturing the callus is 23~27℃, and the time for inducing and culturing the callus is 28~32 days.

[0020] Preferably, the callus tissue is cut into 0.5-1.0 cm pieces before being inoculated into the bud induction medium.

[0021] Preferably, the temperature for bud induction culture is 23~27℃, and the bud induction culture time is 35~45 days.

[0022] Preferably, the temperature for the bud proliferation culture is 23~27℃, and the culture time is 28~32 days.

[0023] Preferably, the temperature for the rooting induction culture is 23~27℃, and the rooting induction culture time is 28~32 days.

[0024] Preferably, the nutrient soil includes peat, coconut coir, perlite and vermiculite; the mass ratio of the peat, coconut coir, perlite and vermiculite is (4~6):(1~3):(1~3):1.

[0025] Compared with the prior art, the present invention has the following beneficial effects: The rapid propagation method of Sedum sarmentosum tissue culture provided by the present invention uses Sedum sarmentosum stem segments as explants, and obtains Sedum sarmentosum regenerated seedlings through callus induction culture, bud induction culture, bud proliferation culture medium, and rooting induction culture. By setting specific culture medium composition and culture conditions at each culture stage, the differentiation rate of callus tissue and the survival rate of regenerated seedlings can be significantly improved. Attached Figure Description

[0026] Figure 1 This represents the situation where stem segments were induced in callus induction medium for 30 days.

[0027] Figure 2 This is the result of callus tissue being cultured in bud induction medium for 40 days.

[0028] Figure 3 This represents the bud proliferation process after 30 days in the bud proliferation medium.

[0029] Figure 4 This shows the plants after 30 days of cultivation in rooting medium.

[0030] Figure 5 This is a photo of the regenerated plant after transplanting. Detailed Implementation

[0031] This invention provides a method for rapid propagation of Sedum sarmentosum via tissue culture, comprising the following steps: 1) using Sedum sarmentosum stem segments as explants, disinfecting and cutting the stem segments, and then inoculating them into a callus induction culture medium to obtain callus tissue; 2) inoculating the obtained callus tissue into a shoot induction culture medium to obtain adventitious shoots; 3) inoculating the obtained adventitious shoots into a shoot proliferation culture medium to obtain regenerated seedlings; 4) inoculating the regenerated seedlings into a rooting culture medium to obtain transplanted seedlings; 5) transplanting the transplanted seedlings into soil for cultivation.

[0032] In this invention, *Sedum aizoon* stem segments are used as explants. After disinfection and cutting, the stem segments are inoculated into a callus induction culture medium to induce callus formation and obtain callus tissue. First, *Sedum aizoon* stem segments are collected, rinsed clean, and then further cleaned. Preferably, the cleaning is done using gauze soaked in detergent for 4-6 minutes; then, the segments are rinsed with running water for 25-35 minutes, preferably 28-32 minutes. After cleaning, the stem segments are disinfected. Preferably, the stem segments are immersed in 70-80% alcohol for 25-35 seconds, rinsed 4-5 times, and then placed in 0.09-0.11 wt% HgCl2 solution for 6-8 minutes, followed by rinsing with water 4-5 times. More preferably, the stem segments are immersed in alcohol for 28-32 seconds, rinsed 4-5 times, and then placed in 0.10 wt% HgCl2 solution for 6.5-7.5 minutes, followed by rinsing with water 4-5 times. After disinfecting the stem segment, the brown wounds at both ends are preferably removed, and then the stem segment is cut. The length of the cut stem segment is preferably 0.45~0.55cm, more preferably 0.5cm. The cut stem segment is inoculated into a callus induction medium for callus induction culture to obtain callus tissue. In this invention, the callus induction medium is preferably MS + 0.45~0.55mg / L NAA + 2.8~3.2mg / L 6-BA, more preferably MS + 0.48~0.52mg / L NAA + 2.9~3.1mg / L 6-BA; even more preferably MS + 0.5mg / L NAA + 3.0mg / L 6-BA. In this invention, the temperature for callus induction culture is 23~27℃, preferably 24~26℃, and the culture time is 28~32 days, preferably 30 days.

[0033] Before being inoculated into the bud induction medium, the callus tissue of this invention is preferably cut into pieces of 0.5-1.0 cm. The obtained callus tissue is then inoculated into the bud induction medium for bud induction culture to obtain adventitious buds. In this invention, the bud induction medium is preferably MS + 0.15-0.25 mg / L NAA + 2.8-3.2 mg / L 6-BA, more preferably MS + 0.18-0.22 mg / L NAA + 2.9-3.1 mg / L 6-BA, and even more preferably MS + 0.2 mg / L NAA + 3.0 mg / L 6-BA. In this invention, the temperature for bud induction culture is 23-27℃, preferably 24-26℃, and the bud induction culture time is preferably 35-45 days, more preferably 38-42 days, and even more preferably 40 days. After bud induction culture, clustered adventitious buds with a height of 2.5-3.5 cm are obtained.

[0034] In this invention, the obtained adventitious buds are inoculated into a bud proliferation medium for bud proliferation culture to obtain regenerated seedlings. The bud proliferation medium is preferably MS + 0.15~0.25 mg / L NAA + 2.8~3.2 mg / L 6-BA, more preferably MS + 0.18~0.22 mg / L NAA + 2.9~3.1 mg / L 6-BA, and even more preferably MS + 0.20 mg / L NAA + 3.0 mg / L 6-BA. The temperature for bud proliferation culture is 23~27℃, preferably 24~26℃, and the culture time is preferably 28~32 days, more preferably 29~31 days, and even more preferably 30 days. After the bud proliferation culture is completed, regenerated seedlings of 2.5~3.5 cm are obtained.

[0035] In this invention, the regenerated seedlings are inoculated into a rooting medium for rooting induction culture to obtain transplanted seedlings. The rooting medium is preferably MS + 0.95~1.05 mg / L IBA, more preferably MS + 1.00 mg / L IBA. The rooting induction culture temperature is 23~27℃, preferably 24~26℃, and the rooting induction culture time is preferably 28~32 days, more preferably 29~31 days, and even more preferably 30 days.

[0036] After the rooting induction culture is completed, the transplanted seedlings are transplanted into nutrient soil for further cultivation. Before transplanting, the seedlings are removed and the residual rooting culture medium on the roots is rinsed off. Preferably, the seedlings are then transplanted into pots of nutrient soil that has been thoroughly watered (water is poured onto the nutrient soil until a small amount of water flows out of the drainage hole at the bottom of the pot, indicating thorough watering). The water-to-nutrient soil mass ratio is preferably 1:(15~17), more preferably 1:16. The nutrient soil comprises peat moss, coconut coir, perlite, and vermiculite; the mass ratio of peat moss, coconut coir, perlite, and vermiculite is (4~6):(1~3):(1~3):1, preferably 5:2:2:1. The seedlings are then cultured at 25±2 ℃.

[0037] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0038] Example 1

[0039] Callus induction medium: MS + 0.50 mg / L NAA + 3.0 mg / L 6-BA;

[0040] Bud induction medium: MS + 0.20 mg / L NAA + 3.0 mg / L 6-BA;

[0041] Shoot proliferation medium: MS + 0.20 mg / L NAA + 3.0 mg / L 6-BA;

[0042] Rooting medium: MS + 1.00 mg / L IBA.

[0043] First, cut young stem segments of *Sedum aizoon*, rinse them thoroughly, then wash them for 5 minutes with gauze soaked in laundry detergent, followed by rinsing with tap water for 30 minutes. Next, soak them in 75% alcohol for 30 seconds, then rinse 4-5 times with sterile water. Finally, place them in a 0.1% HgCl2 solution for 7 minutes, and rinse 4-5 times with sterile water to complete the disinfection process. (When disinfecting in the 75% alcohol and 0.1% HgCl2 solution, gently agitate the beaker containing the disinfectant.)

[0044] The second step is to remove the browned wounds at both ends of the disinfected explant, cut the stem segment into 0.5cm lengths, and inoculate it into callus induction medium for 30 days.

[0045] The third step involves selecting callus tissue at the base of the *Sedum aizoon* stem segment where approximately 0.5 cm of callus tissue can be clearly observed. This is then cut into small pieces of 0.5–1.0 cm and cultured in a bud induction medium for 40 days. Clustered buds approximately 3 cm high are then separated and subcultured, inoculated into a bud proliferation medium, and allowed to proliferate for 30 days.

[0046] The fourth step is to cultivate the test-tube seedlings to about 3cm in height on a rooting medium to induce adventitious roots. Transplant them after 30 days of induction.

[0047] Fifth, take healthy Sedum sarmentosum seedlings with well-developed adventitious roots, open the culture bottle cap for 2 days, then take out the seedlings, wash off the root culture medium, and then transplant them into pots of nutrient soil (peat: coconut coir: perlite: vermiculite = 5:2:2:1) that has been soaked in water for 2 hours in advance, and place them in an environment of 25±2 ℃ for cultivation.

[0048] During the culture process, the induction rate, differentiation rate, contamination rate, and survival rate of callus tissue were statistically analyzed.

[0049] The above culture process was carried out in a sterile culture room at 25±2 ℃, and cultured for 12 h daily under a light intensity of 1500~2000 Lx. The experiment was repeated three times, with 30 explants inoculated for each treatment.

[0050] The experiments showed that MS + 0.50 mg / L NAA + 3.0 mg / L 6-BA was the optimal medium for callus induction, with a callus induction rate of 96.67%; MS + 3.0 mg / L NAA + 0.20 mg / L 6-BA was the optimal medium for callus differentiation, with a callus differentiation rate of 83.33%; MS + 3.0 mg / L NAA + 0.20 mg / L 6-BA was the optimal medium for proliferation, with a proliferation coefficient of 16.13; MS + 1.00 mg / L IBA was the optimal medium for rooting, with a rooting rate of 63.33%; and the survival rate of the tissue culture seedlings obtained in the experiments was over 90% in outdoor culture.

[0051] Experimental Example 1

[0052] Comparison of callus induction rate and contamination rate of different explants

[0053] Take tender stem segments and leaves of Sedum aizoon. Rinse them thoroughly and wash them with gauze soaked in laundry detergent for 5 minutes. Then rinse them with tap water for 30 minutes. After treatment with 75% alcohol for 30 seconds, disinfect them with 0.1% HgCl2 solution for 5, 6, 7, and 8 minutes (when disinfecting in 75% alcohol and 0.1% HgCl2 solution, gently shake the beaker containing the disinfectant) and record them as T1, T2, T3, and T4 respectively. Then rinse them with sterile water 4-5 times, cut off the brown wounds at both ends, cut the stem segments into 0.5 cm lengths, and cut the leaves into 5 mm × 5 mm sizes. Then inoculate the stem segments and leaves into MS medium + 0.5 mg / L NAA + 3.0 mg / L 6-BA medium for callus induction. Simultaneously, the explants, after being thoroughly rinsed, were cleaned with gauze soaked in laundry detergent for 5 minutes, followed by rinsing with running water for 30 minutes. They were then treated with 75% alcohol for 30 seconds (T5) and disinfected in 0.1% HgCl2 solution for 7 minutes (T6). Afterward, they were rinsed 4-5 times with sterile water to remove any brown wounds at both ends. Stem segments were cut into 0.5 cm lengths, and leaves were cut into 5 mm × 5 mm pieces. The stem segments and leaves were then inoculated into MS medium supplemented with 0.5 mg / L NAA and 3.0 mg / L 6-BA for callus induction. At 10 and 30 days post-inoculation, the number of stem segments exhibiting contamination or whitening was counted, and the induction rate, contamination rate, and callus induction rate were statistically analyzed.

[0054] Table 1. Effects of different disinfection methods on explant contamination rates

[0055]

[0056] Note: Different letters after the numerical value indicate significant differences (P<0.05).

[0057] In the experiment, the explants were disinfected with 75% alcohol for 30 seconds, rinsed with sterile water, disinfected with 0.1% HgCl2 solution for 7 minutes, and rinsed with sterile water 4-5 times. After one week, clear signs of growth were observed in the explants, and numerous protrusions appeared on the surface of the explants after 20 days. Further culture revealed a small number of green, spherical protrusions, and adventitious buds appeared around 50 days. The survival rate of the treated explants was high, with a contamination rate of only 15.56% and an induction rate of 78.89%. This demonstrates that the sterilization method is more suitable, and the optimal disinfection method for stem segments is: 75% alcohol disinfection for 30 seconds + sterile water rinsing + 0.1% HgCl2 solution disinfection for 7 minutes + sterile water rinsing 4-5 times.

[0058] The highest induction rate was achieved with stem segments of *Sedum aizoon* (78.89% ± 1.93%), while the highest induction rate was achieved with leaves (60.00% ± 3.33%). Conversely, the lowest contamination rate was found in young stem segments (15.56% ± 1.93%) and leaves (18.89% ± 1.11%). This demonstrates that under the same conditions, the contamination and mortality rates of young stem segments of *Sedum aizoon* were lower than those of leaves, indicating a higher induction rate.

[0059] Experiment Example 2

[0060] Comparison of the induction effects of different callus induction media

[0061] Cleaned young stem segments of Sedum aizoon were treated with 75% alcohol for 30 seconds, then disinfected in 0.1% HgCl2 solution for 7 minutes. After rinsing with sterile water 4-5 times to remove browned wounds at both ends, the stem segments were cut into 0.5 cm lengths and inoculated into MS medium containing different concentrations of NAA (0.1, 0.5 mg / L) and 6-BA (1.0, 2.0, 3.0, 5.0 mg / L) to induce callus tissue. The number of adventitious buds was counted after 30 days.

[0062] In a medium supplemented with 0.5 mg / L NAA and 3.0 mg / L 6-BA, callus growth on the surface of stem explants was rapid. After 30 days of inoculation, 100% of the *Sedum aizoon* explants inoculated in W6 medium showed callus growth, and the callus growth was more vigorous than in other treatments (e.g., ...). Figure 1 (As shown in Table 2). However, under the same culture time, the young stem segments of *Sedum aizoon* inoculated in other media did not show significant changes in the early stages, with small callus volume and slow growth rate. Table 2 shows that the W6 medium had a significantly higher healing rate than the others, making it more suitable for callus production. The optimal hormone ratio for *Sedum aizoon* callus induction culture was: MS + 0.5 mg / L NAA + 3.0 mg / L 6-BA.

[0063] Table 2. Effects of different plant growth regulator concentrations on callus induction in stem segments.

[0064]

[0065] Note: Different letters after the numerical value indicate significant differences (P<0.05).

[0066] Comparison of the effects of different adventitious bud induction media and bud proliferation culture.

[0067] When callus tissue of 0.5 cm in height can be clearly observed at the base of the callus tissue in the stem segment of Sedum sarmentosum, it is cut into small pieces of 0.5-1.0 cm and added to MS basal medium containing different concentrations of NAA (0.1, 0.2, 0.3 mg / L) and 6-BA (1.0, 2.0, 3.0 mg / L) for adventitious shoot induction. The number of newly added adventitious shoots is counted after 30 days.

[0068] Fifteen days after inoculation, most treatments still only produced callus tissue. However, in the MS + 0.2 mg / L NAA + 3.0 mg / L 6-BA treatment, the callus tissue inoculated with *Sedum aizoon* explants showed relatively obvious green buds on the surface, and some even had distinct small green buds. The ends of the stem segments swelled, and adventitious buds began to grow, appearing singly or in clusters, with the highest budding rate of 33.33%. In other treatments, green buds were not clearly observed until 30 days of culture, while at this time, this treatment showed more green spherical bodies on the surface of *Sedum aizoon* callus tissue, and bud differentiation clearly began.

[0069] In MS medium supplemented with 3.0 mg / L 6-BA and 0.2 mg / L NAA, the callus tissue exhibited relatively strong mitotic activity, initially turning green (as shown in the image). Figure 2 As shown in the figure), there are more green spherical bodies, and the adventitious shoot induction rate reaches 83.33%. Therefore, the optimal shoot induction medium for callus differentiation of Sedum aizoon is 0.2 mg / L NAA and 3.0 mg / L 6-BA.

[0070] The clustered buds were separated and passaged, and inoculated into each proliferation medium. The proliferation of the buds was observed after 20 days.

[0071] Test-tube seedlings approximately 3 cm tall were inoculated into different proliferation media containing MS medium supplemented with (0.1-0.3 mg / L) NAA and (1.0-3.0 mg / L) 6-BA. The proliferation coefficients were all above 4.47. The seedlings grown in the medium supplemented with 0.2 mg / L NAA and 3.0 mg / L 6-BA showed the most robust shoot growth and the most vibrant green leaves. Figure 3 The proliferation coefficient was 16.13. Therefore, the optimal bud proliferation medium for Sedum sarmentosum is MS medium supplemented with 0.2 mg / L NAA and 3.0 mg / L 6-BA.

[0072] Table 3. Effects of different hormone concentration ratios on adventitious bud induction in *Sedum morganianum* stem segments.

[0073]

[0074] Note: Different letters after the numerical value indicate significant differences (P < 0.05).

[0075] Comparison of the effects of different rooting media on root induction

[0076] When the test-tube seedlings reached a height of 3 cm, their roots were cultured on MS basal medium with different concentrations of IBA (0, 0.1, 0.2, 0.3, 0.5 mg / L), and the rooting status was recorded.

[0077] The rooting rate of explants on media supplemented with auxin reached 16.67% or higher, while the rooting rate of explants on media without auxin was only 6.67%, with shorter roots and slower growth. When adventitious shoots were transferred to MS medium supplemented with 1.0 mg / L IBA, the rooting rate was 63.33%, with longer and thicker roots and faster growth (e.g., ...). Figure 4 As shown in the figure, low IBA concentrations significantly promote root formation. In conclusion, the optimal rooting medium is determined to be MS + 1.0 mg / L IBA.

[0078] Table 4. Effects of different concentrations of auxin IBA on the rooting of adventitious buds of Sedum aizoon.

[0079]

[0080] Note: Different letters after the numerical value indicate differences (P < 0.05).

[0081] As can be seen from the above embodiments and experimental examples, the rapid propagation method of Sedum sarmentosum tissue culture provided by the present invention can significantly improve the differentiation rate of callus tissue and the survival rate of regenerated seedlings.

[0082] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for rapid propagation of Sedum morganianum via tissue culture with mineral inclusions, characterized in that, Includes the following steps: 1) Using the stem segments of Sedum sarmentosum as explants, the stem segments were disinfected, cut, and then inoculated into callus induction culture medium to induce callus culture and obtain callus tissue. 2) The obtained callus tissue was inoculated into a bud induction medium for bud induction culture to obtain adventitious buds; 3) The obtained adventitious buds are inoculated into bud proliferation medium for bud proliferation culture to obtain regenerated seedlings; 4) The regenerated seedlings are inoculated into a rooting medium for rooting induction culture to obtain transplanted seedlings; 5) Transplant the seedlings into nutrient soil for cultivation; The callus induction culture medium was MS + 0.45~0.55 mg / L NAA + 2.8~3.2 mg / L 6-BA; The bud induction medium was MS + 0.15~0.25 mg / L NAA + 2.8~3.2 mg / L 6-BA; The bud proliferation medium was MS + 0.15~0.25 mg / L NAA + 2.8~3.2 mg / L 6-BA; The rooting medium was MS + 0.95~1.05 mg / L IBA.

2. The method for rapid propagation of Sedum sarmentosum via tissue culture according to claim 1, characterized in that, The stem segment disinfection process involves immersing the stem segments in 70-80% alcohol for 25-35 seconds, rinsing 4-5 times, then placing them in 0.09-0.11wt% HgCl2 solution for 6-8 minutes, and rinsing with water 4-5 times.

3. The rapid propagation method of Sedum sarmentosum tissue culture according to claim 2, characterized in that, Before disinfecting the stem segments, a cleaning step is also included. The cleaning is carried out using gauze soaked in laundry detergent, followed by rinsing with running water for 25-35 minutes.

4. The method for rapid propagation of Sedum sarmentosum via tissue culture according to any one of claims 1 to 3, characterized in that, The length of the cut stem segment is 0.45~0.55cm.

5. The method for rapid propagation of Sedum sarmentosum via tissue culture according to claim 1, characterized in that, The temperature for inducing and culturing the callus is 23~27 ℃, and the induction and culturing time is 28~32 days.

6. The method for rapid propagation of Sedum sarmentosum via tissue culture according to claim 1 or 5, characterized in that, Before being inoculated into the bud induction medium, the callus tissue was cut into pieces of 0.5–1.0 cm.

7. The method for rapid propagation of Sedum sarmentosum via tissue culture according to claim 1 or 5, characterized in that, The temperature for bud induction culture is 23~27℃, and the bud induction culture time is 35~45 days.

8. The method for rapid propagation of Sedum sarmentosum via tissue culture according to claim 7, characterized in that, The bud The temperature for proliferation culture was 23~27℃, and the duration of the bud proliferation culture was 28~32 days.

9. The method for rapid propagation of Sedum sarmentosum via tissue culture according to claim 1, characterized in that, The rooting induction culture temperature is 23~27 ℃, and the rooting induction culture time is 28~32 days.

10. The method for rapid propagation of Sedum sarmentosum via tissue culture according to claim 1, characterized in that, The nutrient soil comprises peat, coconut coir, perlite and vermiculite; the mass ratio of the peat, coconut coir, perlite and vermiculite is (4~6):(1~3):(1~3):1.

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