In-vitro culture and rapid propagation method of chlorophytum comosum

Abstract

The embodiment of the invention discloses an in-vitro culture and rapid propagation method of chlorophytum comosum, and relates to the technical field of plant planting, the method comprises the following steps: selecting a healthy chlorophytum comosum plant as a stock plant to obtain an explant; performing disinfection treatment on the explant twice, performing stem culture and proliferation treatment, and performing rooting culture, seedling hardening and temporary planting after the treatment is completed; after the leaves are matured and new root systems grow out, the seedlings can be transplanted to a field. Compared with the prior art, the application successfully establishes a set of efficient bracketplant seedling tissue culture breeding technical system, not only improves the production efficiency of bracketplant seedlings, but also ensures the quality and growth speed of the seedlings. According to the technology, a large amount of chlorophytum comosum seedlings can be produced in a short time, so that the demand pressure on chlorophytum comosum in the current traditional Chinese medicinal material market is effectively relieved.

Description

Technical Field

[0001] The present application relates to the technical field of plant planting, and in particular to an in vitro culture and rapid propagation method of Chlorophytum comosum. Background Art

[0002] Chlorophytum laxum belongs to the genus Chlorophytum of the Liliaceae family. It is a perennial herb, also known as triangle grass, sparse-flowered Chlorophytum, mountain leek and wild ophiopogon, etc. It likes to grow in moist forests and grasslands at low altitudes and is mainly distributed in Guangdong, Guangxi and Hainan. The leaves of Chlorophytum laxum are arranged in two rows, with a distinct main vein in the middle. The roots are slender and clustered or scattered. Chlorophytum laxum is the source plant of the Chinese medicinal material triangle grass. The whole plant is used as medicine and is a commonly used local herb in South China. It is cool in nature, sweet and slightly bitter in taste, and toxic. It has the effects of anti-inflammatory, analgesic, improving microcirculation, clearing away heat and detoxifying, and reducing swelling and relieving pain. It is often used to treat snake bites and traumatic injuries in Zhongshan and Jiangmen, Guangdong. The medicinal ingredients of Chloromaloside A include steroidal saponins, flavonoids, and polyphenolic compounds, such as Chlorophytoside A, hyoscyproside A, 4',5,7-trihydroxy-6,8-dimethylflavone, stigmasterol, palmitic acid, darutoside, sitosterol, isorientin, and isovitexin. Among them, the steroidal saponin 25-R-spirosta-3,5-dien-12β-ol (1) in the roots of Chloromaloside A has a high cytotoxicity against human nasopharyngeal cancer cells. Chloromaloside A has a wider range of anti-toxicity against human cancer cells, such as lung cancer, epithelial cancer, and breast cancer cells. Chloromaloside A extract also has antioxidant capacity.

[0003] With the continuous deepening of the pharmacognosy research of Chlorophytum comosum, its clinical application will be further promoted. At present, Chlorophytum comosum has not been planted on a large scale and its yield is limited. This is mainly because the endosperm of Chlorophytum comosum is small, the seed coat is hard, the germination rate is low, and seed propagation is difficult. In addition, Chlorophytum comosum is in the wild. With the large-scale use of herbicides in agricultural production, the resources of Chlorophytum comosum are scarce and cannot meet the current market demand. Summary of the invention

[0004] The main purpose of this application is to provide an in vitro culture and rapid propagation method for Chlorophytum comosum, aiming to solve the problem that existing planting technology cannot meet the market demand for Chlorophytum comosum.

[0005] To achieve the above objectives, the technical solutions adopted in the embodiments of the present application are as follows:

[0006] The in vitro culture and rapid propagation method of Chlorophytum comosum comprises the following steps:

[0007] S1. Select a healthy Chlorophytum comosum plant as a mother plant, wash the plant, spread out the leaves and cut them off, completely remove the roots, and retain the stem base as an explant; disinfect the explant twice, and after disinfection, obtain a sterile explant;

[0008] S2, after cutting off the two ends of the stem segment of the sterile explant in contact with the disinfectant, insert the stem segment into an induction medium in an incubator at 28±2°C with the biological lower end facing downward, and first culture it in the dark for 2-3 days, and then culture it in alternating light / darkness with 16h / 8h per day for 5-7 days, and obtain the primary generation material after new leaves sprout;

[0009] S3, take the primary material, cut off the newly extracted leaves, inoculate the sterile explants into the bud proliferation medium for subculture for 20-25 days, and grow lateral buds around the mother stem; when the new leaves are 7 cm long, subculture, cut off the leaves, retain the stem base, and continue to inoculate into the bud proliferation medium for 20-25 days;

[0010] S4. When the leaves of the proliferating buds are against the cap of the culture bottle, cut off a single adventitious bud from the cluster buds, keep a 2-3 cm long leaf, and insert it upright into the rooting medium with the biological bottom facing downward; after 15-20 days, the regenerated plants of Chlorophytum comosum are obtained;

[0011] S5. Take the regenerated plants of Chlorophytum comosum with roots growing to about 1-2cm and seedling height over 7cm for seedling hardening and transplanting; when the leaves mature and new roots grow, they can be transplanted to the field.

[0012] Furthermore, in step S1, the process of performing two disinfection treatments on the explant includes:

[0013] First disinfection: Soak in 70%-75% alcohol for 30 seconds; then soak and wash with sterile water for 3 times, stirring with sterile tweezers during soaking, 5 minutes each time, and dry on sterile paper towels; then transfer to 0.1% mercuric chloride solution containing Tween and soak for 5-11 minutes, stirring with sterile tweezers during soaking, and disinfect thoroughly. Soak and wash the explants with sterile water for 3-5 times, 3 minutes each time;

[0014] Second disinfection: After the explants disinfected for the first time are dried, they are soaked in 500 mg / L Temeiting and 0.1 mg / L GA solution for 20 min.

[0015] Furthermore, in step S2, the induction medium includes MS medium, and the MS medium contains 3.0 mg / L 6-BA, 0.5 mg / L IBA and 0.5 mg / L glucose.

[0016] Furthermore, in step S2, the illuminance is 1900-2100Lx.

[0017] Preferably, in step S1, during the first disinfection, the alcohol concentration of the alcohol soaking is 70%, and the soaking time is 30 seconds; the soaking time in the 0.1% mercuric chloride solution containing Tween is 7-9 minutes.

[0018] Furthermore, the step of taking the regenerated plants of Chlorophytum comosum with roots growing to about 1-2 cm and a seedling height of more than 7 cm for seedling hardening and transplanting comprises:

[0019] Take the regenerated plants of Chlorophytum comosum with roots growing to about 1-2cm and seedling height over 7cm and put them into culture bottles, open the bottle caps of the culture bottles, and harden the seedlings indoors for 5-7 days; take out the tissue culture seedlings, wash the culture medium at the roots with water without damaging the root system; then soak the plants in a solution containing 0.1% carbendazim for 10 minutes, and finally transplant the tissue culture seedlings in a mixture of peat soil and garden soil in a ratio of 3:1, spray water every morning and evening to maintain the humidity of the environment.

[0020] Furthermore, the bud proliferation culture medium includes MS culture medium, and the MS culture medium contains 1.0 mg / L-5.0 mg / L 6-BA and 0 mg / L-0.5 mg / L IBA.

[0021] Preferably, the bud proliferation medium comprises MS medium, and the MS medium contains 3.0 mg / L 6-BA and 0.1 mg / L IBA.

[0022] Furthermore, the rooting medium includes MS medium, and the MS medium contains 0 mg / L-0.5 mg / L NAA and 0 mg / L-0.5 mg / L IBA.

[0023] Preferably, the rooting medium comprises MS medium containing 0.3 mg / L NAA and 0.2 mg / L IBA.

[0024] Compared with the existing technology, this application has deeply studied the main limiting factors faced by Chlorophytum comosum in the process of tissue culture and rapid propagation. Through detailed analysis and research on these limiting factors, we have successfully established a set of efficient Chlorophytum comosum seedling tissue culture and breeding technology system. This technical system not only improves the production efficiency of Chlorophytum comosum seedlings, but also ensures the quality and growth rate of the seedlings. In order to verify the actual application effect of this technical system, we conducted an intermediate test. The test results show that this method can produce a large number of Chlorophytum comosum seedlings in a short period of time, thereby effectively alleviating the current market demand pressure for Chlorophytum comosum seedlings. This achievement not only provides technical support for the large-scale production of Chlorophytum comosum, but also brings new opportunities for the development of related industries. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a diagram showing the results of an explant culture experiment of Chlorophytum comosum involved in an embodiment of the present application;

[0026] Figure 2 This is a graph showing the results of an experiment on adventitious bud induction of Chlorophytum comosum involved in the embodiments of the present application;

[0027] Figure 3 This is a graph showing the results of a Chlorophytum comosum proliferation experiment involved in an embodiment of the present application;

[0028] Figure 4 This is a diagram showing the results of a rooting induction experiment on Chlorophytum comosum involved in an embodiment of the present application;

[0029] Figure 5 This is a graph showing the experimental results of producing bagged seedlings of Chlorophytum comosum involved in the embodiments of the present application;

[0030] Figure 6 This is a diagram of the experimental results of field planting of Chlorophytum comosum involved in the embodiments of the present application. DETAILED DESCRIPTION

[0031] It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0032] Chlorophytum comosum, as a traditional local medicinal plant with a long history, has always attracted much attention for its unique medicinal value. However, due to the particularity and limitations of its growth environment, the natural yield of Chlorophytum comosum is far from meeting the growing market demand. In order to break through this bottleneck, this research team is committed to developing an efficient and rapid Chlorophytum comosum propagation technology.

[0033] The technical solution described in this application is described in detail below in conjunction with specific embodiments:

[0034] It should be noted that the materials used in the following examples and comparative examples are the same, namely:

[0035] Chlorophytum comosum: In the summer of 2022, healthy wild Chlorophytum comosum was selected in Zhongshan, Guangdong, and cultivated in the Key Laboratory of Crop Genetic Improvement of Guangdong Province.

[0036] Induction medium: Add 3.0 mg / L 6-BA, 0.5 mg / L IBA and 0.5 mg / L glucose to MS medium;

[0037] Cluster sprout proliferation medium: add 1.0mg / L-5.0mg / L 6-BA and 0mg / L-0.5mg / L IBA to MS medium;

[0038] Rooting medium: Add 0mg / L-0.5mg / L NAA and 0mg / L-0.5mg / L IBA to MS medium;

[0039] In actual use, 30 g / L sucrose and 5.8 g / L agar were added to the induction medium, bud proliferation medium and rooting medium, the pH value was adjusted to 5.8±0.2, and sterilized at 121°C for 15-20 min.

[0040] Culture conditions: temperature 28±2℃, humidity 65-80%, light intensity 1900-2100Lx, lighting time 12h / d.

[0041] 1) Explant disinfection:

[0042] Select healthy small-flowered Chlorophytum plants as mother plants, wash the plants, especially the soil at the roots, and wipe them dry with clean paper towels. Cut off the unfolded leaves on the clean bench, completely remove the roots, and retain the stem base, about 2 cm long, as explants. The first disinfection uses alcohol and mercuric chloride solution. Soak in 75% alcohol for 30 seconds, then soak and wash with sterile water 3 times. Stir with sterile tweezers during soaking, 5 minutes each time, dry on sterile paper towels, and then transfer to 0.1% mercuric chloride solution containing Tween and soak for 5-11 minutes. The time setting is as shown in Table 1. Stir with sterile tweezers during soaking to fully disinfect. Finally, soak and wash the material with sterile water 3-5 times, 3 minutes each time. Use sterile tweezers to place the treated stem segments on sterile filter paper to dry, and obtain the sterilized explants for use; the second disinfection is to disinfect and dry the explants of treatment group 1-treatment group in Table 1, and continue to soak them in 500 mg / L Temeiting and 0.1 mg / L GA solution for 20 minutes, and record them as composite treatment group 1-composite treatment group 4 respectively; and set the control treatment group for the first disinfection treatment: treatment group 1-treatment group 4 in Table 1; 20 stem segments were disinfected in each treatment group.

[0043] Table 1 Explant disinfection time

[0044]

[0045] 2) Stem segment culture:

[0046] Cut off both ends of the stem segment that was in contact with the disinfectant, cut each end into about 0.2 cm, and insert the stem segment into the induction medium in an incubator at 28±2°C with the biological lower end facing downward. The contamination rate and germination rate of each group after culture, the explants with new leaves sprouting and sterile are the primary generation materials.

[0047] After the explants were sterilized with alcohol and HgCl2, they began to sprout and new leaves grew after being directly cultured on the induction medium for 15 days. During the culture process of the induction medium, they were first cultured in the dark for 2-3 days, and then cultured in alternating light / darkness with 16h / 8h per day for 5 days. Then they began to sprout and new leaves grew (such as Figure 1 The contamination rate, germination rate and mortality rate were statistically analyzed, and the disinfection efficiency of Chlorophytum comosum explants and the germination effect of stem segment culture at different HgCl2 treatment times were analyzed. The results are shown in Table 2.

[0048] Table 2 Effects of different methods on explant disinfection and germination effects of stem segment culture

[0049]

[0050] As shown in Table 2, the contamination of Chlorophytum comosum explants is mainly caused by bacteria. It is possible that there are more endophytes, resulting in incomplete disinfection. The contamination rate of control treatment 4 is 40% lower than that of control treatment 1, but the germination rate of control treatment 4 is only 10%, which is 25% lower than that of control treatment 3; the mortality rate of control treatment 4 is the highest, up to 65%, while that of control treatment 1 is 35%. With the extension of HgCl2 treatment time, the contamination rate of explants decreases, the browning and mortality of explants continue to increase, the germination rate of explants also gradually decreases, and the leaves cultured in stem segments grow out in 12-15 days. Compared with the germination rate of other explants after disinfection, the germination rate of Chlorophytum comosum explants after disinfection is lower. It may be that after the leaves are removed, due to the large gap in the middle of the stem, the HgCl2 solution enters the vicinity of the growth point, and the extension of disinfection time increases the damage to the growth point. In the present application, treatment groups 1-4 can obtain sterile explants. Combined with the comprehensive analysis of germination rate and contamination rate, it can be seen that it is more appropriate to use 70% alcohol for 30s and 0.1% HgCl2 for 7-9 minutes to disinfect the small-flowered Chlorophytum explants; while the contamination rate of composite treatment 1-composite treatment 4 is significantly lower than that of control treatment 1-control treatment 4, the germination rate is significantly increased compared with control treatment 1-control treatment 4, the mortality rate is significantly lower than that of control treatment 1-control treatment 4, and the leaf growth time is significantly shortened compared with control treatment 1-control treatment 4. Therefore, the subsequent embodiments use 70% alcohol for 30s and 0.1% HgCl2 for 7 minutes for disinfection, and then in the process of stem segment cultivation, first dark culture for 3d, and then 16h / 8h light / dark alternating culture for 7d every day.

[0051] 3) Proliferation of buds:

[0052] Take the primary material, cut off the newly drawn leaves, and inoculate the sterile explants into the bud proliferation medium containing different hormone ratios. The hormone ratios are shown in Table 3. Subculture is carried out for 20-25 days, and lateral buds grow around the mother stem. When the new leaves are 7 cm long, subculture is carried out, the leaves are cut off, and the stem base is about 1.5 cm long, and then inoculated into the bud proliferation medium. Continue to culture for 20-25 days, count the number of 20 lateral buds, and compare the efficiency of 6-BA and IBA at different concentrations on the proliferation of buds of Chlorophytum parviflora. The results are shown in Table 4.

[0053] Table 3 Adventitious bud proliferation hormone table

[0054]

[0055] Table 4 Effects of different hormones and concentrations on adventitious bud proliferation

[0056]

[0057] The subculture proliferation culture of adventitious buds can proliferate a large number of Chlorophytum comosum tissue culture seedlings. The level of its proliferation efficiency is the key to the cost of Chlorophytum comosum factory seedling cultivation and determines the application prospects of tissue culture seedlings in production. As can be seen from Table 4, all eight treatment groups can induce adventitious buds of Chlorophytum comosum and proliferate them, but the proliferation efficiency is positively correlated with the concentration of 6-BA, which also shows that Chlorophytum comosum is more sensitive to 6-BA. With the increase of 6-BA concentration in the culture medium, the proliferation efficiency of Chlorophytum comosum tissue culture seedlings also increases. When the 6-BA concentration is 5 mg / L, the proliferation coefficient can reach 7.6, but the buds are too many and weak, which is not conducive to rooting culture (such as Figure 2 and Figure 3 As shown in Figure 2). At the same time, when only 6-BA was used for proliferation culture, the buds were too dense, the seedlings were relatively weak, and the survival rate after rooting culture was not high. Although low concentrations of IBA reduced the proliferation efficiency, the regenerated seedlings were relatively strong, which improved the uniformity of tissue culture seedlings.

[0058] The optimal bud proliferation medium screened out was: 3.0 mg / L 6-BA and 0.1 mg / L IBA were added to MS medium. Therefore, the following examples adopted: 3.0 mg / L 6-BA and 0.1 mg / L IBA were added to MS medium.

[0059] 4) Rooting induction of Chlorophytum comosum:

[0060] When the leaves of the proliferating buds are against the cap of the culture bottle, a single adventitious bud is cut from the cluster buds, and a 2-3 cm long leaf is retained. Then, the biological bottom of the leaf is downward and inserted upright into the rooting medium, about 0.3 cm. Different concentrations of NAA and IBA are added to the MS medium, and the concentrations are shown in Table 5. The rooting induction culture is carried out for 15-20 days, and the number and length of roots of 20 plants are counted. The results are shown in Table 6.

[0061] Table 5 Root inducing hormone table

[0062]

[0063] Table 6 Effects of different hormones on root induction of Chlorophytum comosum

[0064]

[0065]

[0066] After 20 days of rooting induction, the number of roots, root length and state of the tissue culture seedlings were counted. The results showed that the combination of IBA and NAA could induce rooting in the tissue culture seedlings of Chlorophytum comosum, and the induction effect of NAA was better (Table 6). NAA treatment could induce roots, and at the same concentration, the number of roots induced by NAA was greater than that of IBA ( Figure 4). And with the increase of concentration, the number of roots also increased. The number of roots in treatment 2 was 3.16, which was 1.77 times more than that in treatment 1. IBA can promote the thickening of roots and improve the quality of roots. Among them, the number of roots in the treatments of 0.3mg / LNAA and 0.2mg / LIBA was about 3.2, and the number and consistency of roots were good ( Figure 5 ).

[0067] Therefore, the rooting medium in the subsequent examples was: 0.3 mg / L NAA and 0.2 mg / L IBA were added to the MS medium.

[0068] 5) Transplanting and planting of Chlorophytum comosum:

[0069] Take the regenerated plants of the small-flowered spider plant with roots growing to about 1-2cm and a seedling height of more than 7cm for seedling hardening and transplanting. Open the bottle cap of the culture bottle and harden the seedlings indoors for 5-7 days; take out the tissue culture seedlings, wash the culture medium at the roots with water, and do not damage the root system; then soak the plants in a solution containing 0.1% carbendazim for 10 minutes, and finally transplant the tissue culture seedlings in a 3:1 mixture of peat soil and garden soil, spray water every morning and evening to maintain the humidity of the environment. When the leaves are mature and new roots grow, they can be transplanted to the field.

[0070] After 20 days of rooting induction, the roots of the Chlorophytum comosum seedlings generally reach 3 cm in length and the plant height is about 7 cm. After 15 days of transplanting, the leaves are upright, the length, width and thickness increase, and the survival rate is above 97%. The transplanted seedlings are transplanted into the field in spring. After 5 days of transplanting, the leaves of the plants turn green and the survival rate is above 95% ( Figure 6 ).

[0071] In summary, with the deepening of molecular pharmacognosy and pharmacology research, a large number of new active substances have been isolated from medicinal plants, but most of these medicinal plants are in a wild state, and the reserves of resources are low. Once the market is developed, the damage to wild resources will be great. Developing seedling breeding and efficient planting technology for medicinal plant resources is an important basis for protecting and utilizing medicinal plant resources, which is also an important part of the domestication and planting of Chinese medicinal materials. At present, many rare medicinal plants have been artificially domesticated and planted in a large-scale standard manner, such as Dendrobium officinale, Anoectochilus roxburghii, and Bletilla striata, which not only solves the contradiction between the protection of wild resources and the supply of market needs, but also produces good economic benefits. The domestication and planting of Chinese medicinal materials is a favorable guarantee measure for the sustainable development of the traditional Chinese medicine industry. As a traditional folk Chinese herbal medicine, Chlorophytum comosum is one of the important raw materials for the in-hospital preparations of Zhongshan Chinese Medicine Hospital, and it has received more and more attention, but due to the deteriorating site environment, it is in short supply in the market. Establishing the rapid propagation technology of Chlorophytum comosum seedlings can improve the production cost of seedlings, shorten the planting time, save the planting cost, and provide raw material guarantee for the industrialization of Chlorophytum comosum. The rapid propagation technology of medicinal plant tissue can provide a large number of seedlings in a short time, ensuring the large-scale planting of Chlorophytum comosum. At present, the technology of rapid propagation of seedlings using medicinal plant tissue culture technology is mature and widely used. Technically, the acquisition and proliferation efficiency of sterile explants are the key factors in the establishment of tissue culture system, which determines whether the tissue culture rapid propagation technology can be used to produce seedlings of this variety. In production, experienced researchers often combine their own experience to set a simple gradient first, and combine the fine adjustment of parameters to obtain an efficient tissue culture system. Most beginners use orthogonal design to conduct a large number of experiments on the culture medium to screen for a suitable tissue culture system. In plant tissue culture research, it is most common to use HgCl2 as a disinfectant to treat explants. It is also widely used in the tissue culture of Chinese herbal medicines, and the effect is also good, but the morphology and physiological state of the material have a greater impact on the disinfection effect. 0.1% HgCl2 treatment for 7-9 minutes is more suitable for the disinfection of stem segments of Chlorophytum comosum, and then continue to soak in 500mg / L Temeiting and 0.1mg / LGA solution for 20min; during the cultivation of stem segments, first culture them in the dark in the incubator for 3d, and then culture them in alternating light / darkness with 16h / 8h every day for 5-7d to improve the germination rate, thereby greatly shortening the cultivation time of stem segments. Auxin and cytokinin are the main exogenous hormones in plant tissues. Through different content ratios, they induce the proliferation and rooting of explants, and the optimal formula can be obtained through optimization. A large number of plants have established tissue culture technology systems. In the study of establishing tissue culture systems for new species, the results of plants of the same genus can be referred to. The development of the tissue culture system of Chlorophytum comosum, combined with previous research, set different 6-BA concentrations, and obtained a proliferation system suitable for the regeneration of Chlorophytum comosum through the proliferation coefficient and cluster bud quality. NAA and IBA are often used for rooting induction of regenerated plants. Through the gradient combination of content, the root induction system of Chlorophytum comosum was screened.The combination of tissue culture technology, seedling domestication and high-efficiency cultivation technology of medicinal plants can increase the planting scale and efficiency of Chlorophytum comosum and meet the current dilemma of insufficient market demand. It can also provide sufficient raw materials for research and product development, which is beneficial to protecting the wild resources of Chlorophytum comosum and promoting the healthy development of the Chlorophytum comosum industry.

[0072] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A method for in vitro culture and rapid propagation of Chlorophytum comosum, characterized in that: The following steps are involved: S1. Select a healthy Chlorophytum comosum plant as the mother plant, wash the plant, spread out the leaves and cut them off, completely remove the roots, and keep the stem base as the explant; The explant is sterilized twice to obtain a sterile explant; S2, after cutting off the two ends of the stem segment of the sterile explant in contact with the disinfectant, insert the stem segment into an induction medium in an incubator at 28±2°C with the biological lower end facing downward, and first culture it in the dark for 2-3 days, and then culture it in alternating light / darkness with 16h / 8h per day for 5-7 days, and obtain the primary generation material after new leaves sprout; S3, take the primary material, cut off the newly extracted leaves, inoculate the sterile explants into the bud proliferation medium for subculture for 20-25 days, and grow lateral buds around the mother stem; when the new leaves are 7 cm long, subculture, cut off the leaves, retain the stem base, and continue to inoculate into the bud proliferation medium for 20-25 days; S4. When the leaves of the proliferating buds are against the cap of the culture bottle, cut off a single adventitious bud from the cluster of buds, keep a 2-3 cm long leaf, and insert it upright into the rooting medium with the biological bottom facing downward; After 15-20 days, regenerated plants of Chlorophytum comosum were obtained; S5. Take the regenerated plants of Chlorophytum comosum with roots growing to about 1-2cm and seedling height over 7cm for seedling hardening and transplanting; wait until the leaves mature and new roots grow, and then transplant them to the field.

2. The in vitro culture and rapid propagation method of Chlorophytum comosum according to claim 1, characterized in that: In step S1, the process of performing two disinfection treatments on the explant includes: First disinfection: Soak in 70%-75% alcohol for 30 seconds; then soak and wash with sterile water for 3 times, stirring with sterile tweezers during soaking, 5 minutes each time, and dry on sterile paper towels; then transfer to 0.1% mercuric chloride solution containing Tween and soak for 5-11 minutes, stirring with sterile tweezers during soaking, and disinfect thoroughly. Soak and wash the explants with sterile water for 3-5 times, 3 minutes each time; Second disinfection: After the explants disinfected for the first time are dried, they are soaked in 500 mg / L Temeiting and 0.1 mg / L GA solution for 20 min.

3. The in vitro culture and rapid propagation method of Chlorophytum comosum according to claim 1, characterized in that: In step S2, the induction medium includes MS medium, and the MS medium contains 3.0 mg / L 6-BA, 0.5 mg / L IBA and 0.5 mg / L glucose.

4. The in vitro culture and rapid propagation method of Chlorophytum comosum according to claim 1, characterized in that: In the step S2, the illuminance is 1900-2100Lx.

5. The in vitro culture and rapid propagation method of Chlorophytum comosum according to claim 2, characterized in that: In the step S1, the alcohol concentration of the alcohol soaking during the first disinfection is 70%, and the soaking time is 30 seconds; the soaking time in the 0.1% mercuric chloride solution containing Tween is 7 minutes to 9 minutes.

6. The in vitro culture and rapid propagation method of Chlorophytum comosum according to claim 1, characterized in that: The steps of taking the regenerated plants of Chlorophytum comosum with roots growing to about 1-2 cm and a seedling height of more than 7 cm for seedling hardening and transplanting include: Take the regenerated plants of Chlorophytum comosum with roots growing to about 1-2cm and seedling height over 7cm and put them into culture bottles, open the bottle caps of the culture bottles, and harden the seedlings indoors for 5-7 days; take out the tissue culture seedlings, wash the culture medium at the roots with water without damaging the root system; then soak the plants in a solution containing 0.1% carbendazim for 10 minutes, and finally transplant the tissue culture seedlings in a mixture of peat soil and garden soil in a ratio of 3:1, spray water every morning and evening to maintain the humidity of the environment.

7. The in vitro culture and rapid propagation method of Chlorophytum comosum according to claim 1, characterized in that: The bud proliferation culture medium comprises MS culture medium, and the MS culture medium contains 1.0 mg / L-5.0 mg / L of 6-BA and 0 mg / L-0.5 mg / L of IBA.

8. The in vitro culture and rapid propagation method of Chlorophytum comosum according to claim 7, characterized in that: The bud proliferation culture medium comprises MS culture medium, and the MS culture medium contains 3.0 mg / L 6-BA and 0.1 mg / L IBA.

9. The in vitro culture and rapid propagation method of Chlorophytum comosum according to claim 1, characterized in that: The rooting medium includes MS medium, and the MS medium contains 0 mg / L-0.5 mg / L NAA and 0 mg / L-0.5 mg / L IBA.

10. The in vitro culture and rapid propagation method of Chlorophytum comosum according to claim 9, characterized in that: The rooting medium includes MS medium, and the MS medium contains 0.3 mg / L NAA and 0.2 mg / L IBA.

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