A rapid propagation method for Dioscorea alata tissue culture with Dioscorea alata tubers as explants

Abstract

The present invention discloses a rapid propagation method for arrowroot tissue culture with arrowroot tubers as explants, which comprises the following steps: selecting arrowroot tubers as explants and performing disinfection treatment; cutting the disinfected explants into small pieces and inoculating them into an induction medium for cluster bud induction culture; inoculating the induced arrowroot cluster buds into a proliferation medium for proliferation culture; and inoculating the arrowroot tissue culture seedlings obtained by proliferation culture into a rooting medium for rooting culture to obtain rooted seedlings. The present invention uses arrowroot tubers as explants and produces arrowroot seedlings through an in vitro rapid propagation method, which can rapidly and effectively produce a large number of high-quality arrowroot seedlings, is suitable for factory production of seedlings, realizes large-scale rapid propagation of arrowroot, breaks through the traditional ramet propagation method, solves the problem of the proliferation rate of arrowroot, and has important significance for large-scale planting and popularization and utilization of arrowroot.

Description

Technical Field

[0001] The present invention belongs to the technical field of plant tissue culture, and particularly relates to a rapid propagation method for arrowroot tissue culture with arrowroot tubers as explants. Background Art

[0002] Arrowroot, also known as arrowroot powder, scientific name Maranta arundinacea, is a relatively rare ingredient. Its appearance is similar to bamboo shoots, light yellow, 5 to 7 cm long, with white flesh rich in starch, and almost all of its rhizome part is starch.

[0003] Arrowroot belongs to the Marantaceae family, is a perennial herb. Its leaves are long oval, the ground stem is a scaly stem, the flower stalk is slender, the flower is light white, the tuber is round or oval, with a sweet and light taste and a cool nature. It can clear heat and relieve cough, and promote bowel movement.

[0004] Arrowroot tubers are rich in dietary fiber, easy to digest, and can be used for making biscuits and cooking. Eating them can increase the sense of satiety, promote intestinal peristalsis to improve constipation, help with internal environmental protection, beauty care, promote metabolism, detoxify and reduce unnecessary burdens on the body. It is a traditional health care ingredient with the effects of relieving summer heat and fatigue.

[0005] Currently, arrowroot is usually propagated by dividing plants. In a specific season, the mother plant is divided and planted, and generally, some newly grown strong plants are separated from the mother plant as seedlings. However, this propagation method is not only limited by the seasonal climate, but also has a very low multiplication rate, seriously hindering the large-scale rapid propagation of arrowroot and unable to ensure the stability of the horticultural traits of arrowroot. Summary of the Invention

[0006] To solve the above-mentioned drawbacks of the existing technology, the purpose of the present invention is to provide a rapid propagation method for arrowroot tissue culture with arrowroot tubers as explants. The propagation method of the present invention uses tubers for in vitro rapid propagation, solving the problem of low multiplication rate in traditional plant division propagation.

[0007] To achieve the above purpose, the present invention provides the following technical solutions:

[0008] A rapid propagation method for arrowroot tissue culture with arrowroot tubers as explants, comprising the following steps:

[0009] (1) Select arrowroot tubers as explants and perform disinfection treatment;

[0010] (2) Cut the disinfected explants into small pieces and inoculate them into the induction medium for adventitious bud induction culture; the induction medium is: MS+(2.5 - 3.0)mg / L 6 - BA+(1.5 - 2.0)mg / L IBA+(25 - 30)g / L sugar+(7 - 9)g / L carrageenan;

[0011] (3) Inoculate the induced Manihot esculenta Crantz adventitious buds into the proliferation medium for proliferation culture; the proliferation medium is: MS+(2.0 - 2.5)mg / L 6 - BA+(1.0 - 1.5)mg / L IBA+(0.08 - 0.12)mg / L NAA+(25 - 30)g / L sugar+(7 - 9)g / L carrageenan;

[0012] (4) Inoculate the proliferated Manihot esculenta Crantz buds obtained from proliferation culture into the rooting medium for rooting culture, the rooting medium is: MS or 1 / 2MS+(0.08 - 0.12)mg / L NAA+(25 - 30)g / L sugar+(7 - 9)g / L carrageenan+(0.4 - 0.6)g / L activated carbon, to obtain rooted seedlings.

[0013] Preferably, in step (1), the Manihot esculenta Crantz tuber is a mature tuber with bud eyes.

[0014] Preferably, in step (1), the disinfection treatment includes: first wash the sediment on the Manihot esculenta Crantz tuber with water, then soak it in a diluted solution of laundry detergent, then rinse it with running water, then disinfect it with 75% alcohol by volume on the ultra - clean workbench, then disinfect it with 0.1% mercuric chloride solution by mass, and finally wash it with sterile water.

[0015] Preferably, in step (2), the induction medium is: MS + 2.5mg / L 6 - BA + 2.0mg / L IBA + 30g / L sugar + 8g / L carrageenan.

[0016] Preferably, in step (2), the culture temperature for adventitious bud induction culture is 25 - 28°C, and the culture time is 20 - 25 days.

[0017] Preferably, in step (3), the proliferation medium is: MS + 2.0mg / L 6 - BA + 1.5mg / L IBA + 0.1mg / L NAA + 30g / L sugar + 8g / L carrageenan.

[0018] More preferably, coconut water is added to the proliferation medium, and the proliferation medium is: MS + 2.0mg / L 6 - BA + 1.5mg / L IBA + 0.1mg / L NAA + 15mg / L coconut water + 30g / L sugar + 9g / L carrageenan.

[0019] Preferably, in step (3), the conditions for proliferation culture are as follows: culture for 30 - 45 days, culture temperature of 25 - 28°C, light exposure time of 8 - 10 h / day, and light intensity of 2500 - 4000 lx.

[0020] Preferably, in step (3), the number of times of proliferation culture is 3 - 5 times.

[0021] Preferably, the rooting medium is: 1 / 2MS + 0.1 mg / L NAA + 25 g / L sugar + 9 g / L carrageenan + 0.5 g / L activated carbon.

[0022] Preferably, the conditions for rooting culture are as follows: culture for 45 - 60 days, culture temperature of 26 - 30°C, light exposure time of 8 - 10 h / day, and light intensity of 2500 - 4000 lx.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0024] (1) In the present invention, the arrowroot tuber is used as the explant, and arrowroot seedlings are produced by an in vitro rapid propagation method. By optimizing and screening the induction medium for the explant, the proliferation medium for clustered buds, and the rooting medium for tissue culture seedlings, the induction rate of the explant reaches more than 86%, the proliferation coefficient of the clustered buds reaches more than 4.3, and the rooting rate of the tissue culture seedlings reaches more than 100%. The present invention can rapidly and effectively produce a large number of high-quality arrowroot seedlings, is suitable for factory production of seedlings, realizes large-scale rapid propagation of arrowroot, breaks through the traditional ramet propagation method, solves the problem of the proliferation rate of arrowroot, and has important significance for the large-scale planting and popularization and utilization of arrowroot.

[0025] (2) In the present invention, a certain amount of fresh coconut water is added to the proliferation medium, which effectively improves the proliferation coefficient of the clustered buds and further improves the effect of rapid propagation of arrowroot tissue. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a diagram of the arrowroot tuber explant of Example 1 of the present invention;

[0027] Figure 2 It is a diagram of the induced arrowroot clustered buds of Example 1 of the present invention;

[0028] Figure 3 It is a diagram of the proliferated arrowroot clustered buds of Example 1 of the present invention;

[0029] Figure 4 It is a diagram of the rooted arrowroot seedlings of Example 1 of the present invention during rooting culture. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0030] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0031] The sugar used in the embodiments of the present invention is sucrose.

[0032] The activated carbon used in the embodiments of the present invention is 200-mesh activated carbon powder.

[0033] The coconut water used in the embodiments of the present invention is the coconut water taken out after breaking the shell of fresh coconuts (fruit age 11 - 12 months).

[0034] Example 1

[0035] A rapid propagation method for Dioscorea alata tissue culture with tubers as explants, which comprises the following steps:

[0036] (1) Selection and disinfection of explants: Select mature Dioscorea alata tubers with bud eyes as explants, wash the sand on the tubers with tap water, then soak them in a diluted solution of laundry detergent for 30 min, then rinse with running water for 30 min, then disinfect them with 75% alcohol by volume for 30 s on a super clean bench, and then disinfect them with 0.1% mercuric chloride by mass for 7 min, and finally wash them with sterile water 3 - 4 times;

[0037] (2) Induction of cluster buds: Cut the disinfected explants into small pieces, inoculate them into the induction medium for cluster bud induction culture for 20 days, and the culture temperature is 25 - 28 °C; the induction medium is: MS + 2.5 mg / L 6 - BA + 2.0 mg / L IBA + 30 g / L sugar + 8 g / L carrageenan;

[0038] (3) Proliferation culture of cluster buds: Inoculate the induced Dioscorea alata cluster buds into the proliferation medium for proliferation culture for 30 days, the culture temperature is 25 - 28 °C, the light time is 10 h / day, and the light intensity is 2500 lx; the proliferation medium is: MS + 2.0 mg / L 6 - BA + 1.5 mg / L IBA + 0.1 mg / L NAA + 30 g / L sugar + 9 g / L carrageenan;

[0039] (4) Rooting culture: Inoculate the Dioscorea alata proliferation buds obtained by proliferation culture 5 times into the rooting medium for rooting culture for 50 days, the culture temperature is 26 - 30 °C, the light time is 10 h / day, and the light intensity is 2500 lx to obtain rooted seedlings; the rooting medium is 1 / 2MS + 0.1 mg / L NAA + 25 g / L sugar + 9 g / L carrageenan + 0.5 g / L activated carbon.

[0040] Example 2

[0041] A rapid propagation method for cassava tissue culture with tuber as explant, which comprises the following steps:

[0042] (1) Selection and disinfection of explant: Select mature cassava tubers with bud eyes as explants, wash the sand on the tubers with tap water, then soak them in a diluted solution of laundry detergent for 30 min, then rinse with running water for 30 min, then disinfect with 75% (v / v) alcohol for 30 s on a super clean bench, then disinfect with 0.1% (w / v) mercuric chloride solution for 7 min, and finally wash with sterile water 3 - 4 times;

[0043] (2) Induction of cluster buds: Cut the disinfected explants into small pieces, inoculate them into the induction medium for cluster bud induction culture for 20 days, and the culture temperature is 25 - 28 °C; The induction medium is: MS + 3.0 mg / L 6 - BA + 2.0 mg / L IBA + 30 g / L sugar + 8 g / L carrageenan;

[0044] (3) Multiplication culture of cluster buds: Inoculate the induced cassava cluster buds into the multiplication medium for multiplication culture for 30 days, the culture temperature is 25 - 28 °C, the light time is 9 h / day, and the light intensity is 3000 lx; The multiplication medium is: MS + 2.5 mg / L 6 - BA + 1.5 mg / L IBA + 0.1 mg / L NAA + 30 g / L sugar + 9 g / L carrageenan; The multiplication coefficient reaches 2.3;

[0045] (4) Rooting culture: Inoculate the cassava proliferated buds obtained by 5 - time multiplication culture into the rooting medium for rooting culture for 50 days, the culture temperature is 26 - 30 °C, the light time is 9 h / day, and the light intensity is 3000 lx to obtain rooted seedlings; The rooting medium is: MS + 0.1 mg / L NAA + 30 g / L sugar + 9 g / L carrageenan + 0.5 g / L activated carbon.

[0046] Example 3

[0047] The main difference from Example 1 is that coconut water is also added to the multiplication medium.

[0048] A rapid propagation method for cassava tissue culture with tuber as explant, which comprises the following steps:

[0049] (1) Selection and disinfection of explant: Select mature cassava tubers with bud eyes as explants, wash the sand on the tubers with tap water, then soak them in a diluted solution of laundry detergent for 30 min, then rinse with running water for 30 min, then disinfect with 75% (v / v) alcohol for 30 s on a super clean bench, then disinfect with 0.1% (w / v) mercuric chloride for 7 min, and finally wash with sterile water 3 - 4 times;

[0050] (2) Induction of clustered buds: Cut the disinfected explants into small pieces, inoculate them into the induction medium for 20 days of induction culture of clustered buds, and the culture temperature is 25 - 28°C; the induction medium is: MS + 2.5 mg / L 6 - BA + 2.0 mg / L IBA + 30 g / L sugar + 8 g / L carrageenan;

[0051] (3) Proliferation culture of clustered buds: Inoculate the induced clustered buds of cassava into the proliferation medium for 30 days of proliferation culture, the culture temperature is 25 - 28°C, the light time is 10 h / day, and the light intensity is 2500 lx; the proliferation medium is: MS + 2.0 mg / L 6 - BA + 1.5 mg / L IBA + 0.1 mg / L NAA + 15 mg / L coconut water + 30 g / L sugar + 9 g / L carrageenan;

[0052] (4) Rooting culture: Inoculate the proliferated buds of cassava obtained by 5 - time proliferation culture into the rooting medium for 50 days of rooting culture, the culture temperature is 26 - 30°C, the light time is 10 h / day, and the light intensity is 2500 lx to obtain rooted seedlings; the rooting medium is 1 / 2MS + 0.1 mg / L NAA + 25 g / L sugar + 9 g / L carrageenan + 0.5 g / L activated carbon.

[0053] Comparative Example 1

[0054] The main difference from Example 1 is that the concentration of 6 - BA in the induction medium and the proliferation medium is different. A rapid propagation method for tissue culture of cassava using tubers as explants includes the following steps:

[0055] (1) Selection and disinfection of explants: Select mature cassava tubers with bud eyes as explants, wash the sand on the tubers with tap water, then soak them in a diluted solution of laundry detergent for 30 min, then rinse them with running water for 30 min, then disinfect them with 75% alcohol by volume for 30 s on the ultra - clean workbench, then disinfect them with a 0.1% mercuric chloride solution by mass for 7 min, and finally wash them 3 - 4 times with sterile water;

[0056] (2) Induction of clustered buds: Cut the disinfected explants into small pieces, inoculate them into the induction medium for 20 days of induction culture of clustered buds, and the culture temperature is 25 - 28°C; the induction medium is: MS + 2.0 mg / L 6 - BA + 2.0 mg / L IBA + 30 g / L sugar + 8 g / L carrageenan;

[0057] (3) Proliferation culture of cluster buds: The induced cluster buds of arrowroot were inoculated into the proliferation medium for 30 days of proliferation culture. The culture temperature was 25 - 28 °C, the light time was 8 h / day, and the light intensity was 4000 lx. The proliferation medium was: MS + 3.0 mg / L 6 - BA + 1.5 mg / L IBA + 0.1 mg / L NAA + 30 g / L sugar + 9 g / L carrageenan;

[0058] (4) Rooting culture: The arrowroot proliferation buds obtained after 3 times of proliferation culture were inoculated into the rooting medium for 50 days of rooting culture. The culture temperature was 26 - 30 °C, the light time was 8 h / day, and the light intensity was 4000 lx to obtain rooted seedlings. The rooting medium was: 1 / 2MS + 0.1 mg / L NAA + 25 g / L sugar + 9 g / L carrageenan + 0.5 g / L activated carbon.

[0059] Comparative Example 2

[0060] The main difference from Example 1 is that the concentration of 6 - BA in the induction medium and the proliferation medium is different. A rapid propagation method for arrowroot tissue culture with tubers as explants includes the following steps:

[0061] (1) Selection and disinfection of explants: Select mature arrowroot tubers with bud eyes as explants. Wash the sand on the tubers with tap water, then soak them in a diluted solution of laundry detergent for 30 min, then rinse with running water for 30 min, then disinfect them with 75% (v / v) alcohol for 30 s on a super clean bench, and then disinfect them with 0.1% (w / v) mercuric chloride solution for 9 min, and finally wash them 3 - 4 times with sterile water;

[0062] (2) Induction of cluster buds: Cut the disinfected explants into small pieces and inoculate them into the induction medium for 20 days of cluster bud induction culture. The culture temperature was 25 - 28 °C. The induction medium was: MS + 3.5 mg / L 6 - BA + 2.0 mg / L IBA + 30 g / L sugar + 8 g / L carrageenan;

[0063] (3) Proliferation culture of cluster buds: The induced cluster buds of arrowroot were inoculated into the proliferation medium for 30 days of proliferation culture. The culture temperature was 25 - 28 °C, the light time was 10 h / day, and the light intensity was 2500 lx. The proliferation medium was: MS + 1.5 mg / L 6 - BA + 1.5 mg / L IBA + 0.1 mg / L NAA + 30 g / L sugar + 9 g / L carrageenan;

[0064] (4) Rooting culture: The multiplied bamboo potato buds obtained after 5 times of multiplication culture were inoculated into the rooting medium for 50 days of rooting culture at a culture temperature of 26 - 30°C, a light duration of 10 h / day, and a light intensity of 2500 lx to obtain rooted seedlings; the rooting medium was: 1 / 2MS + 0.1 mg / L NAA + 25 g / L sugar + 9 g / L carrageenan + 0.5 g / L activated carbon.

[0065] Comparative Example 3

[0066] The main difference from Example 3 was the different addition amounts of coconut water in the multiplication medium.

[0067] A rapid propagation method for bamboo potato tissue culture using tubers as explants, which includes the following steps:

[0068] (1) Selection and disinfection of explants: Select mature bamboo potato tubers with bud eyes as explants, wash the sand on the tubers with tap water, then soak them in a diluted solution of laundry detergent for 30 min, then rinse with running water for 30 min, then disinfect them with 75% (v / v) alcohol for 30 s on a super clean bench, and then disinfect them with 0.1% (w / v) mercuric chloride for 7 min, and finally wash them 3 - 4 times with sterile water;

[0069] (2) Induction of cluster buds: Cut the disinfected explants into small pieces and inoculate them into the induction medium for 20 days of cluster bud induction culture at a culture temperature of 25 - 28°C; the induction medium was: MS + 2.5 mg / L 6 - BA + 2.0 mg / L IBA + 30 g / L sugar + 8 g / L carrageenan;

[0070] (3) Multiplication culture of cluster buds: Inoculate the induced bamboo potato cluster buds into the multiplication medium for 30 days of multiplication culture at a culture temperature of 25 - 28°C, a light duration of 10 h / day, and a light intensity of 2500 lx; the multiplication medium was: MS + 2.0 mg / L 6 - BA + 1.5 mg / L IBA + 0.1 mg / L NAA + 60 mg / L coconut water + 30 g / L sugar + 9 g / L carrageenan;

[0071] (4) Rooting culture: The multiplied bamboo potato buds obtained after 5 times of multiplication culture were inoculated into the rooting medium for 50 days of rooting culture at a culture temperature of 26 - 30°C, a light duration of 10 h / day, and a light intensity of 2500 lx to obtain rooted seedlings; the rooting medium was 1 / 2MS + 0.1 mg / L NAA + 25 g / L sugar + 9 g / L carrageenan + 0.5 g / L activated carbon.

[0072] Test Example

[0073] The experiments were carried out according to Examples 1-3 and Comparative Examples 1-3, which were Experimental Groups 1-6 in sequence. There were 100 small pieces of explants for each experimental treatment. The number of adventitious buds formed was recorded, the induction rate of explants in each experimental group was calculated, the number of new bud proliferations was recorded, the proliferation coefficient of adventitious buds was calculated, and the rooting rate of tissue culture seedlings was calculated. The results are shown in Table 1.

[0074] The induction rate of explants = the number of explants forming adventitious buds / the total number of inoculated explants * 100%;

[0075] The proliferation coefficient of adventitious buds = the number of proliferated buds / the total number of inoculated adventitious buds;

[0076] The rooting rate = the number of tissue culture seedlings with roots / the total number of Manihot esculenta Crantz tissue culture seedlings * 100%.

[0077] Table 1 Tissue culture situation of Manihot esculenta Crantz

[0078]

[0079]

[0080] In the present invention, the tuber of Manihot esculenta Crantz is used as the explant, and the seedlings of Manihot esculenta Crantz are produced by in vitro rapid propagation method. By optimizing and screening the induction medium for explants, the proliferation medium for adventitious buds and the rooting medium for tissue culture seedlings, the induction rate of explants can reach more than 86%, the proliferation coefficient of adventitious buds can reach more than 4.3, and the rooting rate of tissue culture seedlings can reach more than 100%, and the root growth vitality is strong (the average root length of Examples 1-3 is 10-15 cm).

[0081] Among them, in Comparative Examples 1-2, the concentration of 6-BA in the induction medium for explants and the proliferation medium for adventitious buds was adjusted, and the induction rate of explants and the proliferation coefficient of adventitious buds decreased significantly, indicating that adding a certain concentration of 6-BA combined with IBA and NAA in the present invention effectively improves the induction and proliferation effects of the induction medium for explants and the proliferation medium for adventitious buds.

[0082] In Example 3 of the present invention, a certain amount of coconut water was added to the proliferation medium, which effectively improved the proliferation coefficient of adventitious buds. Among them, compared with Example 3, in Comparative Example 3, more coconut water was added, but the increase amplitude of the proliferation coefficient of adventitious buds decreased instead.

[0083] Although the present invention has been described in detail with reference to the foregoing embodiments, for those skilled in the art, they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for tissue culture propagation of bamboo yam using bamboo yam tubers as explants, characterized in that: The steps include: (1) Selecting bamboo shoot tubers as explants and disinfecting them; (2) cutting the sterilized explants into small pieces and inoculating them into an induction medium for cluster bud induction culture; the induction medium comprises: MS + (2.5-3.0) mg / L 6-BA + (1.5-2.0) mg / L I BA + (25-30) g / L sugar + (7-9) g / L carrageenan; (3) inoculating the induced bamboo shoots into a proliferation medium for proliferation culture; the proliferation medium comprises: MS + (2.0-2.5) mg / L 6-BA + (1.0-1.5) mg / L I BA + (0.08-0.12) mg / L NAA + (25-30) g / L sugar + (7-9) g / L carrageenan; (4) inoculating the bamboo shoots obtained by proliferation culture into a rooting medium for rooting culture, wherein the rooting medium is: MS or 1 / 2MS + (0.08-0.12) mg / L NAA + (25-30) g / L sugar + (7-9) g / L carrageenan + (0.4-0.6) g / L activated carbon to obtain rooted seedlings; The bamboo yam tuber is a mature bamboo yam tuber with buds.

2. The method for tissue culture and propagation of bamboo yam using bamboo yam tubers as explants according to claim 1, characterized in that: Step (2), the induction medium is: MS+2.5mg / L6-BA+2.0mg / L IBA+30g / L sugar+8g / L carrageenan.

3. The method for tissue culture and propagation of bamboo yam using bamboo yam tubers as explants according to claim 1 or 2, characterized in that: Step (2), the culturing temperature of the clustered bud induction culture is 25-28°C, and the culturing time is 20-25 days.

4. The method for tissue culture and propagation of bamboo yam using bamboo yam tubers as explants according to claim 1, characterized in that: Step (3), the proliferation medium is: MS+2.0mg / L6-BA+1.5mg / L IBA+0.1mg / L NAA+30g / L sugar+8g / L carrageenan.

5. A method for tissue culture propagation of bamboo yam using bamboo yam tubers as explants as claimed in claim 1, characterized in that: Replace the proliferation medium in step (3) with: MS + 2.0 mg / L 6-BA + 1.5 mg / L IBA + 0.1 mg / L NAA + 15 mg / L coconut water + 30 g / L sugar + 9 g / L carrageenan.

6. The method for tissue culture propagation of bamboo yam using bamboo yam tubers as explants according to claim 1 or 5, characterized in that: Step (3), the conditions of the proliferation culture are: culture for 30-45 days, culture temperature of 25-28°C, illumination time of 8-10h / day, and illumination intensity of 2500-4000lx.

7. The method for tissue culture and propagation of bamboo yam using bamboo yam tubers as explants according to claim 1, characterized in that: In step (3), the proliferation culture is performed 3-5 times.

8. The method for tissue culture and propagation of bamboo yam using bamboo yam tubers as explants according to claim 1, characterized in that: The rooting medium is: 1 / 2MS+0.1mg / LNAA+25g / L sugar+9g / L carrageenan+0.5g / L activated carbon.

9. The method for tissue culture propagation of bamboo yam using bamboo yam tubers as explants according to claim 1 or 8, characterized in that: The rooting culture conditions are: culture for 45-60 days, culture temperature of 26-30° C., light duration of 8-10 h / day, and light intensity of 2500-4000 lx.

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