A tissue culture rapid propagation method for Feige Blue potted flowers

Abstract

The present invention "a tissue culture rapid propagation method of the blue pot flower" belongs to the field of plant tissue culture rapid propagation technology, and is characterized in that it includes the following steps: S1, preparing a culture medium; the culture medium includes: MS culture medium, callus induction culture medium, callus differentiation and proliferation culture medium and rooting induction culture medium; S2, inducing callus culture; inoculating an explant sample into the callus culture medium until callus tissue is formed; S3, callus differentiation and proliferation; inoculating an explant with callus tissue into the callus differentiation and proliferation culture medium, and cultivating it to form a bud cluster; S4, inducing rooting culture; inoculating the bud cluster into the rooting induction culture, and performing rooting cultivation; S5, transplanting tissue seedlings; after the bud cluster is greater than 4 cm in height, transplanting it into a substrate, ensuring the environmental humidity, and performing normal cultivation; the substrate is a mixture of peat soil, perlite, and sand in a volume ratio of 4:3:2.

Description

Technical Field

[0001] The present application belongs to the technical field of plant tissue culture rapid propagation, and specifically relates to a tissue culture rapid propagation method of the Flying Pigeon Blue Pot Flower. Background Art

[0002] Scabiosa columbaria 'Butterfly Blue' is a perennial herb and an important flower species. Its vibrant blue-purple flowers, beautifully shaped, and long-lasting blooms make it a highly ornamental plant, suitable for use in potted plants, flower beds, and other garden settings. The plant also has medicinal properties. In Europe and Africa, its leaves, roots, and the entire plant are used to treat respiratory ailments and skin infections.

[0003] In horticultural production, the excellent characteristics of the Flying Pigeon Blue Flower cannot be maintained through seed propagation. Currently, the main method of preserving the excellent germplasm is cutting propagation. However, in practice, Flying Pigeon Blue Flower cuttings are difficult to root and have low propagation efficiency, which seriously limits the preservation of Flying Pigeon Blue Flower germplasm, market supply, and industrial development. Tissue culture rapid propagation systems may be a potential solution to this problem.

[0004] In terms of existing tissue culture technology, there are only a few technical literatures that conduct preliminary research on this.

[0005] Xu Dongting et al. (Tissue Culture and Plant Regeneration of Scabiosa columbaria, Acta Botanica Sinica, 2011, 46(4): 476-478; DOI: 10.3724 / SP.J.1259.2011.00476) and patent CN114208673A (A Tissue Culture Method for Scabiosa columbaria) provided some basic data but did not propose specific treatment methods and lacked technical support. Romeijn and Andr'e AM van Lammeren (Plant regeneration through callusinitiation from anthers and ovules of Scabiosa columbaria, Plant cell, tissue and organ culture, 1999, 56: 169-177) only analyzed the proliferation of gamete cells of Scabiosa columbaria by in vitro culture of anthers and ovules.

[0006] Takashi Hosoki and Sachie Nojima ("Micropropagation of Scabiosa caucasicabieb. cv. Caucasica blue," In Vitro Cellular & Developmental Biology-Plant; 2004, 40:482-484.) studied explants of Scabiosa caucasica, including stem tips, leaves, and petioles, and found that 6-benzyladenine (6-BA) and indolebutyric acid (IBA) played a significant role in tissue culture. In summary, rapid tissue culture propagation technology for Scabiosa caucasica remains largely undeveloped, with no specific treatment methods or culture systems specifically tailored to this plant. Further research is urgently needed to achieve rapid propagation and industrial application. Summary of the Invention

[0007] In order to address the gaps in the existing technology, the present invention has explored and tried the culture conditions of callus induction, callus differentiation and proliferation, rooting induction and other links in the tissue culture process of Feige Blue Potted Flower one by one, and obtained a set of efficient and systematic Feige Blue Potted Flower tissue culture rapid propagation system.

[0008] Specifically, the scheme of the present invention is as follows:

[0009] A tissue culture rapid propagation method for the Flying Pigeon Blue pot flower, characterized by comprising the following steps:

[0010] S1. Prepare culture medium:

[0011] MS basal medium: Dissolve 4.74 g / L of MS powder without agar and sucrose, 30 g / L of sucrose, and 7 g / L of carrageenan in water, adjust the pH to 5.8-6, and sterilize at 121°C for 20 minutes.

[0012] Callus induction medium: 2.0 mg / L phenylthiadiazolyl urea and 0.5 mg / L α-naphthaleneacetic acid were added to the MS basal medium;

[0013] The callus differentiation and proliferation medium is the same as the MS basal medium;

[0014] The root induction medium is prepared by adding 2.0 mg / L of indolebutyric acid to MS medium;

[0015] S2. Callus induction: inoculate the explant sample of the Flying Pigeon Blue Pot Flower into the callus induction medium to culture and form callus;

[0016] S3. Callus differentiation and proliferation: inoculate the callus tissue into callus differentiation and proliferation medium and cultivate it until adventitious buds are differentiated;

[0017] S4. Root induction: The adventitious buds were inoculated into a root induction medium and cultured to grow into sterile seedlings;

[0018] S5. Organize seedling transplanting: When the sterile seedlings are greater than 4 cm in height, transplant them into the substrate and carry out conventional cultivation.

[0019] Preferably, in the tissue culture rapid propagation method, step S1 further comprises:

[0020] S21. Disinfection of explant samples: Select disease-free leaf samples or petiole samples of the Flying Pigeon Blue Pot Plant. Rinse for 30 minutes, then disinfect with 75% alcohol for 30 seconds twice, rinse with sterile water, disinfect with 2% sodium hypochlorite for 9 minutes, and rinse again with sterile water. Drain and cut into pieces for later use.

[0021] Preferably, in the tissue culture rapid propagation method, the callus differentiation and proliferation culture medium does not contain 6-benzyladenine.

[0022] Preferably, in the tissue culture rapid propagation method, the explant sample of the Flying Pigeon Blue Flower is the leaf tissue of the Flying Pigeon Blue Flower cut into a size of 1 cmX1 cm; or the petiole of the Flying Pigeon Blue Flower cut into a length of 1 cm.

[0023] Preferably, in the tissue culture rapid propagation method, the culture conditions for inducing callus tissue in S2 are: culturing at 25±2° C. for 30 days until callus tissue is formed.

[0024] Preferably, in the tissue culture rapid propagation method, the cultivation conditions for callus differentiation and proliferation in S3 are: culturing at 25±2°C until adventitious buds are formed.

[0025] Preferably, in the tissue culture rapid propagation method, the culture conditions for inducing rooting in S4 are: culturing at 25±2°C for 15-20 days until the bud cluster grows roots.

[0026] The invention provides a tissue culture rapid propagation method for the Flying Pigeon Blue Pot Flower, and proposes a specific and efficient technical solution to address the problems of low propagation efficiency, vague processing methods and lack of specific technical guidance in the prior art.

[0027] The present invention achieves callus induction rates as high as 92.38% (petioles) and 86.67% (leaves) by optimizing the ratio of plant growth regulators (e.g., TDZ 2.0 mg / L + NAA 0.5 mg / L). The present invention also significantly improves callus differentiation rate and adventitious bud proliferation number by creatively omitting 6-BA from the differentiation and proliferation medium (see Table 3 and Table 4 of the Examples). Figure 3During the rooting culture stage, IBA (2.0 mg / L) was added to promote root growth, ensuring the rooting rate and plant survival rate, avoiding complex environmental control, and further reducing production difficulty;

[0028] During the seedling transplanting stage, the use of a mixed matrix of peat soil, perlite and sand significantly improved the survival rate of the seedlings.

[0029] The present invention provides a complete, systematic and standard method for rapid tissue culture propagation of the Flying Pigeon Blue Flower, which significantly improves production efficiency. It is not only suitable for large-scale seedling cultivation in horticultural production, but can also be used for the subculture preservation of excellent Flying Pigeon Blue Flower germplasm, providing effective technical support for the commercial supply and medicinal industry development of the Flying Pigeon Blue Flower group, and has significant economic and social benefits. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is a graph showing the effects of different concentrations of plant hormones on the induction of petiole callus;

[0031] Figure 2 This is a graph showing the effects of different concentrations of plant hormones on the induction of leaf callus.

[0032] Figure 3 This is a diagram showing the effects of different concentrations of 6-BA in inducing callus differentiation and adventitious bud proliferation;

[0033] Figure 4 This is the result graph of the effect of different concentrations of IBA on rooting;

[0034] Figure 5 This is a picture of the growth of transplanted seedlings in Shenzhong. DETAILED DESCRIPTION

[0035] To further clarify the objectives, technical solutions, and advantages of this application, the following exemplary descriptions of the technical solutions of this application are provided in conjunction with the following embodiments and accompanying drawings. Those skilled in the art will appreciate that these exemplary descriptions are intended to support the technical solutions claimed in this application and are not intended to limit the scope of protection.

[0036] The following is combined with Figure 1-5 The tissue culture rapid propagation method of the Feige Blue potted flower of the present application is further described in detail.

[0037] The present application discloses a tissue culture rapid propagation method of the Flying Pigeon Blue Flower, wherein the materials are selected from the leaves and petioles of the Flying Pigeon Blue Flower obtained from the Institute of Vegetables and Flowers of the Chinese Academy of Agricultural Sciences, and the method specifically comprises the following steps:

[0038] S1. Prepare the culture medium. The culture medium includes: MS medium, callus induction medium, callus differentiation and proliferation medium, and rooting induction medium. MS basal medium: Mix 4.74 g / L of MS powder (without agar and sucrose), 30 g / L of sucrose, and 7 g / L of carrageenan, adjust the pH to 5.8-6, and sterilize in a horizontal autoclave at 121°C for 20 minutes.

[0039] Callus induction medium: prepared by adding plant growth regulators to the MS basic culture;

[0040] Callus differentiation and proliferation medium: Same as MS basal medium, that is, 4.74 g / L of MS powder without agar and sucrose, 30 g / L of sucrose, and 7 g / L of carrageenan are mixed, the pH is adjusted to 5.8-6, and the mixture is sterilized in a horizontal autoclave at 121°C for 20 minutes.

[0041] Root induction medium: prepared by adding plant growth regulators to the MS basic culture.

[0042] The plant growth regulators used in this study include indolebutyric acid (IBA), α-naphthaleneacetic acid (NAA), phenylthiadiazolyl urea (TDZ), or 6-benzyladenine (6-BA).

[0043] S21. Sample disinfection. Select disease-free plant leaf and / or petiole samples, clean the dust on the leaf surface with tap water and detergent, and rinse under running tap water for 30 minutes. After rinsing, pour the sample into a beaker, move it to the clean bench, and perform the following disinfection steps in sequence: disinfect with 75% alcohol for 30 seconds, repeat twice, and then rinse thoroughly with sterile water to remove alcohol residue; then, soak the sample in 2% sodium hypochlorite solution for 9 minutes for deep disinfection, and rinse again with sterile water to remove disinfectant residue; finally, gently dry the surface of the sample with sterile filter paper, cut the leaves into 1cm*1cm explants; cut the petioles into 1cm long explants for later use.

[0044] S22. Callus induction culture. Disinfected explants were inoculated into callus induction media of varying proportions, each supplemented with different concentrations of plant growth regulators (see Tables 1 and 2 for specific ingredients and proportions). Seven culture flasks were set up for each medium, with five explants inoculated per flask. Culture was maintained at a constant temperature of 25 ± 2°C for 30 days, until callus tissue formed. Periodically photographed during the culture process, and the callus induction rate was calculated at the end of the culture period.

[0045] S3. Callus differentiation and proliferation. Explants containing callus tissue were cut into small pieces of uniform size and inoculated into callus differentiation and proliferation medium with different compositions (see Table 3 for specific compositions and ratios). Eight culture flasks were set up for each medium composition, with three callus explants inoculated into each flask. Culture was carried out at 25 ± 2°C for approximately 60 days until bud clusters formed. Periodic photographs were taken during the culture process. At the end of the experiment, the differentiation rate and the number of adventitious buds differentiated from each callus piece were counted.

[0046] S4. Root induction culture. Select well-growing, uniformly sized adventitious bud clusters and inoculate them in rooting medium with different ratios (see Table 4 for specific ingredients and ratios). For each medium ratio, set up 10 culture bottles, inoculating three bud clusters per bottle. Incubate at 25 ± 2°C for approximately 30 days, until the bud clusters have formed a complete root system and the plant height reaches at least 4 cm. Take photos regularly during the culture process, and record the rooting rate, root length, and number of roots.

[0047] S5. Organize seedling transplanting. When the bud clusters exceed 4 cm in height, select well-rooted seedlings and transplant them into a medium composed of peat moss, perlite, and sand in a 4:3:2 volume ratio. After transplanting, water thoroughly and place the seedlings in a greenhouse, maintaining an ambient temperature of 20-25°C and a humidity of 80%-90%, for normal cultivation. After 15 days of cultivation, observe the growth of the seedlings and calculate the survival rate.

[0048] Experiment and result analysis

[0049] 1. Effects of different concentrations of plant growth regulators on petiole callus induction

[0050] As shown in Table 1, the disinfected petioles were inoculated into MS-based culture medium, and plant growth regulators of different concentrations and combinations were added: TDZ (0.00, 1.00, 2.00 mg / L), TDZ (1.00, 2.00 mg / L) + NAA (0.5 mg / L), 6-BA (0.8, 1.00, 2.00 mg / L) + NAA (0.5 mg / L). Swelling appeared around the wound on about 7 days, and light green callus was induced at the edge of the wound. The callus grew rapidly within 30 days, and a large amount of callus appeared. The callus had different states and colors, including yellow-green, green, and dark green.

[0051] Table 1: Effects of different concentrations of plant growth regulators on the induction of petiole callus

[0052]

[0053] like Figure 1As shown in the figure, the observation results after 3 days of culture showed that no callus tissue could be induced in the MS medium without adding any hormones; when only TDZ was added, although there was a certain callus induction effect, the callus tissue formed was small; when TDZ and NAA were used in combination, the callus induction effect was the best, and the callus tissue formed was dark green in color and compact in structure. Among them, A6 medium (i.e. MS medium + TDZ 2.00 mg / L + NAA 0.05 mg / L) had the best effect of inducing petiole callus; in contrast, when 6-BA and NAA were used in combination, the callus induction effect was the worst, and the callus tissue produced was small and prone to browning.

[0054] Table 1 also shows that when TDZ was used alone, the callus induction rate gradually increased with increasing TDZ concentrations within the 0.00 mg / L to 1.50 mg / L range. When TDZ was used in combination with NAA, the induction effect was significantly enhanced, with the A6 medium achieving the highest induction rate of 92.38%. In contrast, the callus induction effect was poor in the combination of 6-BA and NAA, with induction rates below 40%. The control group, A1 (which did not contain any plant growth regulators), had an induction rate of 0, indicating no callus formation.

[0055] In summary, the optimal culture medium for petiole callus induction was: MS powder (without agar and sucrose) 4.74 g / L, sucrose 30 g / L, carrageenan 7 g / L, TDZ 2.0 mg / L, and NAA 0.5 mg / L.

[0056] 2. Effects of different concentrations of plant growth regulators on leaf callus induction

[0057] As shown in Table 2, the disinfected leaves were inoculated into MS-based culture medium, and different concentrations of TDZ (0.00, 1.00, 2.00 mg / L), TDZ (2.00 mg / L) + NAA (0.5 mg / L), and 6-BA (0.8, 1.00, 2.00 mg / L) + NAA (0.5 mg / L) were added. Around the 7th day, swelling appeared around the wound of the leaf, and light green callus tissue was induced at the edge of the wound.

[0058] Table 2: Effects of different concentrations of plant growth regulators on leaf callus induction

[0059]

[0060] like Figure 2As shown in the table and the statistical data in Table 2, after 30 days of culture, no callus could be induced without the addition of any plant growth regulators. While TDZ alone showed some callus induction, the resulting calli were small, and some showed adventitious bud differentiation. The combination of TDZ and NAA significantly enhanced callus induction. The medium containing 4.74 g / L B5:MS powder (without agar and sucrose), 30 g / L sucrose, 7 g / L carrageenan, 2.0 mg / L TDZ, and 0.5 mg / L NAA was the most effective for callus induction in petioles, with a maximum induction rate of 86.67%. In contrast, the medium containing both 6-BA and NAA exhibited poor callus induction, resulting in smaller calli that were prone to browning, with induction rates below 10%. Increasing 6-BA concentrations increased callus browning, further reducing the induction efficiency.

[0061] In summary, the optimal culture medium for callus induction from leaves was: MS powder (without agar and sucrose) 4.74 g / L, sucrose 30 g / L, carrageenan 7 g / L, TDZ 2.0 mg / L, and NAA 0.5 mg / L.

[0062] 3. Effects of different concentrations of 6-BA on callus differentiation and proliferation

[0063] Calluses inoculated into different callus differentiation and proliferation media were observed to produce adventitious buds in all groups, but the extent of differentiation and proliferation varied greatly. The callus darkened in color around day 10, and adventitious buds sprouted after day 20. After switching to the same culture medium on day 30, the adventitious buds rapidly proliferated by day 60, forming numerous clusters of buds.

[0064] like Figure 3 As shown, the callus differentiation and proliferation of group C1 was the best, and the adventitious buds grew well, were dark green in color, and were tall. With the increase of 6-BA concentration, the color of the adventitious buds after callus differentiation and proliferation turned yellow to a certain extent, and most of the adventitious buds were short.

[0065] As shown in Table 3, the highest callus differentiation and proliferation induction rate was achieved in Group C1, which used a basal medium without 6-BA, achieving an induction rate of 82.22%. Under these conditions, each callus produced over 10 adventitious buds, demonstrating strong differentiation capacity. In contrast, the poorest differentiation effect was observed in Group C5, which, at a 6-BA concentration of 4.00 mg / L, achieved an induction rate of only 11.76%, with fewer than three adventitious buds produced per callus. The overall trend showed that the induction rate of callus differentiation and proliferation decreased significantly with increasing 6-BA concentration.

[0066] Table 3: Effects of different concentrations of 6-BA on callus differentiation and adventitious bud proliferation

[0067]

[0068] In summary, the optimal culture medium for inducing callus differentiation and proliferation is the culture medium without 6-BA, with the specific formula being: MS powder (without agar and sucrose) 4.74 g / L, sucrose 30 g / L, and carrageenan 7 g / L.

[0069] 4. Effects of different concentrations of IBA on rooting culture

[0070] like Figure 4 As shown, well-grown, uniformly sized adventitious buds were inoculated into rooting-inducing medium with varying concentrations of IBA. Roots successfully formed in all treatments. Rooting occurred approximately three days earlier in group D3 than in the other two groups. By day 30 of culture, all buds showed varying degrees of growth, with an increased number of roots and further elongation. Observations revealed that the root system in group D3 was longer, the rhizome was relatively robust, and rooting efficiency and growth vigor were superior to those in the other groups.

[0071] As shown in Table 4, the rooting rates of sterile seedlings in the three different root induction media were all high. When the IBA concentration in the culture medium was 1.00 mg / L and 2.00 mg / L, the rooting rate of sterile seedlings was the highest, reaching 93.33%. Among them, when the IBA concentration was 2.00 mg / L, the average number of roots of sterile seedlings was 9.10, and the proportion of plants with roots longer than 4 cm reached 86.67%, which was significantly higher than the other two groups. Comprehensive analysis of rooting rate and root growth showed that the appropriate addition of IBA can effectively promote the rooting induction of adventitious buds and significantly increase the number of roots and root length.

[0072] Table 4: Effects of different concentrations of IBA on rooting

[0073]

[0074] In summary, the optimal culture medium for root induction was: MS powder (without agar and sucrose) 4.74 g / L, sucrose 30 g / L, carrageenan 7 g / L and IBA 2.0 mg / L.

[0075] 5. Organize seedling transplanting test

[0076] like Figure 5 As shown, sterile seedlings with well-developed root systems and a height greater than 4 cm were removed from the conical flask. After cleaning the remaining culture medium from the roots, the seedlings were transplanted into the substrate and watered thoroughly. After 16 days of incubation, the plants were in good condition, with green leaves and strong growth. A total of 25 seedlings were transplanted, with a survival rate of 100%.

[0077] 6. Final conclusion of the experiment

[0078] The optimal medium for inducing petiole and leaf callus was 4.74 g / L MS powder (without agar and sucrose), 30 g / L sucrose, 7 g / L carrageenan, 2.0 mg / L TDZ, and 0.5 mg / L NAA. In this medium, the callus induction rate for petioles was 92.38%, and for leaves was 86.67%. During the culture process, callus began to appear on the petioles and leaves around the 10th day and entered a rapid growth phase after the 20th day. Its color gradually changed from light green to dark green, and its tissue structure gradually became compacted from loose, indicating good differentiation potential. Furthermore, the callus grew evenly without browning, making it suitable for subsequent adventitious bud differentiation.

[0079] The optimal medium for inducing callus differentiation and proliferation consisted of 4.74 g / L MS powder (without agar and sucrose), 30 g / L sucrose, and 7 g / L carrageenan, without the addition of any plant growth regulators. In this medium, the callus differentiation induction rate reached 82.22%, with an average of 11.8 adventitious buds per callus. Adventitious buds began to appear after the 30th day. These buds were bright green and uniform in size. Over 90% of the buds were in good condition, showing no signs of browning or necrosis, demonstrating excellent growth vitality.

[0080] The optimal culture medium for root induction consisted of 4.74 g / L MS powder (without agar and sucrose), 30 g / L sucrose, 7 g / L carrageenan, and 2.0 mg / L IBA. Under these conditions, the rooting rate of adventitious buds was 93.33%, with an average of 9.10 roots. 86.67% of plants had roots exceeding 4 cm in length. During the culture process, adventitious buds in the D3 group began to root on the 12th day, approximately three days earlier than the other groups. The rooted roots were robust, with abundant root hairs and numerous branches. The root system was a healthy milky white color, demonstrating high vitality and providing a high-quality root foundation for subsequent transplanting.

[0081] The transplanting medium consisted of peat moss, perlite, and sand in a volume ratio of 4:3:2. For transplanting, robust seedlings with well-developed root systems and a height exceeding 4 cm were cleaned of the root culture medium and transplanted into the medium. After being well-watered, the seedlings were incubated in a greenhouse (20-25°C, 80%-90% humidity) for 16 days. A total of 25 seedlings were transplanted in the experiment, with a 100% survival rate. Observation 16 days after transplanting revealed that all seedlings were in good condition, with new leaves sprouting and continued root growth. Their leaves were green and healthy, with no signs of wilting or yellowing, demonstrating strong environmental adaptability and vigor, validating the effectiveness and reliability of this rapid propagation method.

[0082] The above description is merely a specific embodiment of the present application, but the scope of protection of the present application is not limited thereto. Any changes or substitutions that can be easily conceived by a person skilled in the art within the technical scope disclosed in this application should be included in the scope of protection of the present application. Therefore, the scope of protection of the present application should be based on the scope of protection of the claims.

Claims

1. A tissue culture rapid propagation method for the bluebell flower, characterized in that: The following steps are involved: S1. Prepare culture medium: MS basal medium: Dissolve 4.74 g / L of MS powder without agar and sucrose, 30 g / L of sucrose, and 7 g / L of carrageenan in water, adjust the pH to 5.8-6, and sterilize at 121°C for 20 minutes. Callus induction medium: 2.0 mg / L phenylthiadiazolyl urea and 0.5 mg / L α-naphthaleneacetic acid were added to the MS basal medium; Callus differentiation and proliferation medium: Same as MS basal medium, but without 6-benzyladenine; The root induction medium was prepared by adding 2.0 mg / L indolebutyric acid to MS medium; S2. Callus induction: inoculate the explant sample of the Flying Pigeon Blue Pot Flower into the callus induction medium to culture and form callus; S3. Callus differentiation and proliferation: The callus tissue is inoculated into callus differentiation and proliferation medium and cultured until adventitious buds are differentiated; S4. Root induction: The adventitious buds were inoculated into a root induction medium and cultured to grow into sterile seedlings; S5. Organize seedling transplanting: When the sterile seedlings are greater than 4 cm in height, transplant them into the substrate and carry out conventional cultivation.

2. The tissue culture rapid propagation method according to claim 1, wherein step S2 further comprises: S21. Disinfection of explant samples: Select disease-free leaf samples or petiole samples of the Flying Pigeon Blue Pot Plant. Rinse for 30 minutes, then disinfect with 75% alcohol for 30 seconds twice, rinse with sterile water, disinfect with 2% sodium hypochlorite for 9 minutes, and rinse again with sterile water. Drain and cut into pieces for later use.

3. tissue culture rapid propagation method according to claim 1, described Fei Ge Blue Potted Flower explant sample is to be cut into Fei Ge Blue Potted Flower leaf tissue of 1cmX1cm size; Or be cut into Fei Ge Blue Potted Flower petiole of 1cm length.

4. The tissue culture rapid propagation method according to claim 1, wherein the culture conditions for inducing callus in S2 are: culturing at 25±2°C for 30 days until callus is formed.

5. The tissue culture rapid propagation method according to claim 1, wherein the cultivation conditions for callus differentiation and proliferation in S3 are: culturing at 25±2°C until adventitious buds are formed.

6. The tissue culture rapid propagation method according to claim 1, wherein the culture conditions for inducing rooting in S4 are: culturing at 25±2°C for 15-20 days until the bud cluster grows roots.

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