Method for inducing regenerated plant based on small spike-stalks of green-stalk bamboos

Abstract

The invention discloses a method for inducing a regenerated plant based on a small spike stalk of bamboo-stalk, and relates to the technical field of regenerated plants.The method comprises the steps that the small spike of flowering bamboo-stalk is taken, disinfected, washed with sterile water and placed on sterile filter paper to absorb moisture, and dissected to obtain the small spike stalk as an explant; the small spike stalks are inoculated into a callus induction culture medium and cultured for 1-2 weeks under the dark condition, callus is formed through induction, and the induction culture medium comprises an MS culture medium, 2, 4-D, KT, ferulic acid, aspartic acid, Yb, an amino acid mixture and cane sugar; the callus is inoculated into a differential culture medium and cultured under the illumination condition, adventitious buds are differentiated, and the rooting culture medium comprises a 1 / 2 MS culture medium, IBA, NAA, ferric citrate, beta-salicylic acid and cane sugar. According to the method disclosed by the invention, a callus induction system starting from reproductive tissues of the bamboo-stalk bamboo is established, regenerated plants are successfully obtained, and a technical means is provided for rapid propagation of bamboo-stalk bamboo seedlings.

Description

Technical Field

[0001] The invention relates to the technical field of regenerated plants, in particular to a method for inducing regenerated plants based on spikelet axes of green-stalked bamboo. Background Art

[0002] Green-stalked bamboo, also known as green-pole bamboo, is a species of bamboo belonging to the genus Bambusa in the Poaceae family (Bambusa tuldoides Munro). It is 6 to 10 m tall and 3 to 5 cm in diameter. The bamboo stalk is cylindrical and grooved. There is a circle of gray-white hair above and below the first or second sheath ring at the base. It is mostly distributed in Fujian, Yunnan, Guangdong and Guangxi, and often grows on hillsides or on both sides of streams and rivers. Green-stalked bamboo is an economical and practical plant resource. Its stalk has good toughness and ductility, and can be used as a building material for houses and processed into bamboo crafts. The dried middle layer of the bamboo stalk after removing the outer skin can be used as medicine. The Chinese medicine name is "Zhu Ru", which can clear away heat and resolve phlegm, relieve restlessness and stop vomiting. In theory, green-stalked bamboo can be bred by traditional methods such as burying whips, burying stalks or secondary branch cuttings, but these methods have the disadvantages of large seed source requirements, inconvenient transportation, and high labor intensity. Plant tissue culture has the advantages of large reproduction coefficient, rapid growth, disease removal and rejuvenation, and has been widely used in the cultivation of bamboo seedlings. Node buds, anthers and embryos are commonly used explant materials for bamboo plant tissue culture. Among them, node buds are suitable for large-scale reproduction and preservation of excellent varieties, but not for fundamental improvement of varieties; while anther and embryo breeding can achieve variety innovation and genetic improvement by introducing new genetic variations. Summary of the invention

[0003] In view of the above existing problems, the present invention is proposed.

[0004] Therefore, the present invention provides a method for inducing regeneration of plants based on spikelet axes of green-stalked bamboo to solve the shortcomings in existing bamboo plant propagation technologies.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions: In a first aspect, an embodiment of the present invention provides a method for inducing plant regeneration based on spikelet axis of green-stalked bamboo, comprising taking spikelets of flowering green-stalked bamboo, disinfecting them, washing them with sterile water and placing them on sterile filter paper to absorb water, and dissecting the spikelet axis to obtain it as an explant; The spikelet axis is inoculated into a callus induction medium, and cultured in the dark for 1-2 weeks to induce callus formation, wherein the induction medium components include MS medium, 2,4-D, KT, ferulic acid, aspartic acid, Yb, amino acid mixture and sucrose; Inoculating the callus tissue into a differentiation medium, culturing under light conditions, and differentiating adventitious buds, wherein the rooting medium components include 1 / 2MS medium, IBA, NAA, ferric citrate, β-salicylic acid, and sucrose; The adventitious buds are transferred to a rooting medium, and after culturing for a certain period of time, regenerated seedlings with thicker root systems are selected for seedling hardening, and transplanted to a suitable cultivation medium. After survival, they are placed in a greenhouse for growth. The components of the differentiation medium include MS medium, 6-BA, NAA, caproic acid, adenine sulfate and sucrose.

[0006] As a preferred embodiment of the method for inducing regeneration of plants based on spikelet axes of green bamboo described in the present invention, the concentration of 2,4-D in the callus induction medium is 2-5 mg / L, the concentration of KT is 0.2-1 mg / L, the concentration of sucrose is 20-30 g / L, the concentration of ferulic acid is 0.02-0.05 mg / L, the concentration of aspartic acid is 0.05-0.15 mg / L, the concentration of Yb is 3-5 μM, and the concentration of the amino acid mixture is 0.3-0.5 g / L.

[0007] As a preferred embodiment of the method for inducing regeneration of plants based on the spikelet axis of green bamboo described in the present invention, the concentration of 6-BA in the differentiation medium is 4-8 mg / L, the concentration of NAA is 0.2-1 mg / L, the concentration of caproic acid is 0.1-0.5 mg / L, the concentration of adenine sulfate is 20-30 mg / L, and the concentration of sucrose is 20-30 g / L.

[0008] As a preferred embodiment of the method for inducing plant regeneration based on spikelet axes of green-stalked bamboo described in the present invention, the spikelet axis explant is at least one of the base spikelet axis, top spikelet axis or middle spikelet axis of flowering green-stalked bamboo.

[0009] As a preferred embodiment of the method for inducing regeneration of plants based on the spikelet axis of green bamboo described in the present invention, the concentration of IBA in the rooting medium is 0.05-0.15 mg / L, the concentration of NAA is 0.5-1.5 mg / L, the concentration of sucrose is 15-25 g / L, the concentration of ferric citrate is 0.2-1 mg / L, and the concentration of β-salicylic acid is 0.03-0.05 mg / L.

[0010] As a preferred embodiment of the method for inducing plant regeneration based on spikelet axis of green bamboo described in the present invention, the induction culture step is carried out at a temperature of 24-28°C and the culture time is 1-2 weeks.

[0011] As a preferred embodiment of the method for inducing plant regeneration based on the spikelet axis of green bamboo described in the present invention, the rooting culture step of the regenerated seedlings is carried out at a temperature of 22-26°C and the culture time is 4-6 weeks.

[0012] As a preferred embodiment of the method for inducing plant regeneration based on spikelet axis of green-stalked bamboo described in the present invention, the induction culture medium components also include plant peptides and circular RNA.

[0013] As a preferred embodiment of the method for inducing plant regeneration based on spikelet axes of green bamboo described in the present invention, during the treatment of the spikelet axis explants, the mercuric chloride disinfection time is 6-8 minutes, and the spikelet axis explants are rinsed with sterile water 5 times.

[0014] The beneficial effects of the present invention are as follows: a callus induction system starting from the reproductive tissue of green-stalked bamboo is established, and regenerated plants are successfully obtained, thereby providing a technical means for the rapid propagation of green-stalked bamboo seedlings. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.

[0016] Figure 1 This is a schematic diagram of the effects of spikelets of different lengths and different concentrations of 2,4-D on the callus induction rate of spikelet axis of green bamboo. DETAILED DESCRIPTION

[0017] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the accompanying drawings.

[0018] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0019] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0020] Example 1 is the first example of the present invention, which provides a method for inducing plant regeneration based on spikelet axis of green bamboo, comprising the following steps: This embodiment also provides a method for inducing plant regeneration based on spikelet axis of green bamboo, comprising: Take the spikelets of flowering green bamboo, disinfect them, rinse them with sterile water, place them on sterile filter paper to absorb the water, and dissect to obtain the spikelet axis as explant; The spikelet axis is inoculated into a callus induction medium and cultured in the dark for 1-2 weeks to induce callus formation. The induction medium components include MS medium, 2,4-D, KT, ferulic acid, aspartic acid, Yb, amino acid mixture and sucrose, which optimizes the callus induction stage. The culture medium can effectively reduce the oxidative stress of plant cells in the early stage and enhance the vitality of cells, thereby accelerating the formation and proliferation of callus. The addition of Ytterbium (Yb) ions can significantly improve the induction efficiency of callus and provide a more stable basis for the regeneration of bamboo species; The callus tissue is inoculated into a differentiation medium and cultured under light conditions to differentiate adventitious buds. The rooting medium components include 1 / 2MS medium, IBA, NAA, ferric citrate, β-salicylic acid and sucrose. The medium effectively promotes the rapid differentiation and healthy development of adventitious buds. Hexanoic acid can regulate the balance of plant hormones, avoid excessive bias of bud differentiation towards the root system or branches, and ensure the normal development of buds, while the addition of adenine sulfate increases the quantity and quality of adventitious buds. The adventitious buds are transferred to a rooting medium, and after a certain period of cultivation, regenerated seedlings with thicker roots are selected for hardening, and transplanted to a suitable cultivation medium. After survival, they are placed in a greenhouse for growth. The differentiation medium components include MS medium, 6-BA, NAA, caproic acid, adenine sulfate and sucrose. The medium effectively promotes the healthy growth of the root system, reduces oxidative damage, and significantly improves the development speed and quality of the root system. Ferric citrate can increase the absorption of nutrients by the root system, and β-salicylic acid, as an antioxidant, effectively reduces root growth problems caused by environmental stress.

[0021] Furthermore, the concentration of 2,4-D in the callus induction medium is 2-5 mg / L, the concentration of KT is 0.2-1 mg / L, the concentration of sucrose is 20-30 g / L, the concentration of ferulic acid is 0.02-0.05 mg / L, the concentration of aspartic acid is 0.05-0.15 mg / L, the concentration of Yb is 3-5 μM, and the concentration of the amino acid mixture is 0.3-0.5 g / L; the concentration of 6-BA in the differentiation medium is 4-8 mg / L, the concentration of NAA is 0.2-1 mg / L, the concentration of hexanoic acid is 0.1-0.5 mg / L, the concentration of adenine sulfate is 20-30 mg / L, and the concentration of sucrose is 20-30 g / L.

[0022] Preferably, the explant of the spikelet axis is at least one of the base spikelet axis, the top spikelet axis or the middle spikelet axis of the flowering green bamboo, the concentration of IBA in the rooting medium is 0.05-0.15 mg / L, the concentration of NAA is 0.5-1.5 mg / L, the concentration of sucrose is 15-25 g / L, the concentration of ferric citrate is 0.2-1 mg / L, and the concentration of β-salicylic acid is 0.03-0.05 mg / L.

[0023] It should be noted that the induction culture step is carried out at a temperature of 24-28°C for 1-2 weeks, and the rooting culture step of the regenerated seedlings is carried out at a temperature of 22-26°C for 4-6 weeks.

[0024] The induction medium also includes plant peptides and circular RNA, which can effectively promote information transmission between plant cells, enhance the responsiveness of hormone signals, and enhance the induction and stability of callus. These exogenous factors can theoretically enhance the stress response ability of plant cells and provide signal support in the early stage of callus, ultimately increasing the differentiation rate of adventitious buds.

[0025] Preferably, during the treatment of the spikelet axis explants, the mercuric chloride disinfection time is 6-8 minutes, and the spikelet axis explants are rinsed with sterile water 5 times.

[0026] Example 2, reference Figure 1 This embodiment provides a specific implementation method of a method for inducing plant regeneration based on spikelet axis of green bamboo.

[0027] Obtaining and inoculating explants: Take the clustered spikelets with branches of flowering green-stalked bamboo, wipe the surface of the spikelets gently with 75% alcohol for disinfection, and place them at 4 ℃ overnight; select the spikelets with full development and tightly closed florets, grade them according to their length (grade I: less than 15 mm; grade II: 15-25 mm; grade III: more than 25 mm), and then disinfect them separately; disinfect them with 75% alcohol for 30-45 s, rinse them with sterile water 3 times, and then use 0.2% mercuric chloride for shock disinfection for 6-8 min, rinse them with sterile water 5 times and set aside, use sterile gauze to cover the bottle mouth to filter the disinfectant and rinse solution; place the disinfected spikelets on sterile filter paper to absorb moisture, dissect the spikelets with pointed tweezers in both hands, remove the sterile florets at the base and top, dissect the 3-4 usable florets in the middle, and peel out the spikelet axis as explants for later use.

[0028] (2) Callus induction culture: Spikelets of different lengths were inoculated into the induction medium. The callus induction medium was supplemented with 4.405 g / L MS, 0-5 mg / L 2,4-D, 0.5 mg / L KT, 25 g / L sucrose, 7 g / L agar, 0.05 mg / L ferulic acid (Apigenin), 0.1 mg / L aspartic acid (Aspartic Acid), and 0.5 g / L plant peptide (Peptone); the pH was adjusted to 5.8-6.2; the induction conditions were: dark culture, temperature 25 ± 1 ℃. Callus was formed after 2 weeks of induction culture.

[0029] (3) Callus differentiation culture: Inoculate the callus that grows to more than 5 mm, milky white, dense granular, in the differentiation medium. The callus differentiation medium is supplemented with 4.405 g / L MS, 2-6 mg / L 6-BA, 0.1-1 mg / L NAA; 20 mg / L adenine sulfate, 30 g / L sucrose, 7 g / L agar, 0.1 mg / L hexanoic acid; the pH is adjusted to 5.8-6.2; light culture, light intensity 27μmol m-2 s-1, 16 hours light / 8 hours dark photoperiod, temperature 25 ± 1 ℃; after 2-4 weeks of culture, the callus surface turns green and green buds appear; after 8-12 weeks of continuous culture, the green buds differentiate into adventitious buds, and then the buds and leaves gradually expand.

[0030] (4) Rooting culture of regenerated seedlings: Inoculate the well-growing regenerated seedlings into the rooting medium. The rooting medium was supplemented with 1 / 2 MS, 0.1 mg / L IBA, 1.0 mg / L NAA, 25 g / L sucrose, 7 g / L agar, 1 mg / L ferric citrate, and 0.05 mg / L β-salicylic acid; the pH was adjusted to 5.8-6.2; light culture was performed with a light intensity of 27 μmol m-2 s-1, a photoperiod of 16 h light / 8 h dark, and a temperature of 25 ± 1 ℃.

[0031] (5) Hardening and transplanting of regenerated plants: After one month of rooting culture, select the regenerated seedlings with good growth and strong root system for hardening and transplanting. First, place the regenerated seedlings in a half-opened tissue culture bottle for 2-3 days, then remove the culture medium and fill it with tap water. The seedlings are placed in the bottle for another day. At this time, the temperature of the culture room is 25 ± 1℃, the light intensity is 27 μmol m-2 s-1, and the photoperiod is 16 hours of light / 8 hours of darkness; the hardened seedlings are soaked in 0.25% carbendazim for 5 minutes, and transplanted to a culture medium with a ratio of perlite to humus of 1:1. Use white transparent film or plastic bags for shade, maintain a relative air humidity of about 70%, and remove the bags after 2 weeks.

[0032] The spikelet axis of green bamboo was inoculated into the medium with concentration gradient of 1, 2, 3, 4, 5 mg / L 2, 4-D and 0.5 mg / L KT, and the callus induction rate was counted. The concentration of sucrose in this callus induction medium was 20 g / L, the concentration of ferulic acid was 0.02 mg / L, the concentration of aspartic acid was 0.05 mg / L, the concentration of Yb was 3μM, and the concentration of amino acid mixture was 0.3 g / L. It can be seen from Table 1 that 2 mg / L and 3 mg / L 2, 4-D had good callus induction effect on the spikelet axis of green bamboo I (spikelet length less than 15 mm) and II (spikelet length 15-25 mm). Considering the induction rate of spikelets of III (spikelet length more than 25 mm), 3 mg / L 2, 4-D was the best concentration for spikelet callus induction.

[0033] Table 1 Effects of spikelets of different lengths and different concentrations of 2,4-D on the callus induction rate of spikelet axis of green bamboo Spikelet length 2，4-D Callus induction rate% Level I 1 mg / L <![CDATA[57.78±3.80 ab ]]> 2 mg / L <![CDATA[64.44±2.10 ab ]]> 3 mg / L <![CDATA[66.67±3.33 a ]]> 4 mg / L <![CDATA[51.11±2.99 bc ]]> 5 mg / L <![CDATA[42.22±3.55 c ]]> Level II 1 mg / L <![CDATA[35.56±3.46 b ]]> 2 mg / L <![CDATA[66.67±3.88 a ]]> 3 mg / L <![CDATA[64.44±2.86 a ]]> 4 mg / L <![CDATA[40.00±3.24 b <!-- 4 -->]]> 5 mg / L <![CDATA[55.56±3.22 a ]]> Grade III 1 mg / L <![CDATA[8.89±3.47 b ]]> 2 mg / L <![CDATA[13.33±3.56 b ]]> 3 mg / L <![CDATA[28.89±3.55 a ]]> 4 mg / L <![CDATA[33.33±3.66 a ]]> 5 mg / L <![CDATA[31.11±3.79 a ]]> In Table 1, for spikelet length, level I means less than 15 mm, level II means less than 15-25 mm, and level III means more than 25 mm; different lowercase letters indicate significant differences between different concentrations at the same level (P < 0.05).

[0034] The spikelet axis of green bamboo was inoculated into the medium with concentration gradient of 2, 3 mg / L 2, 4-D, 0.02 m, 0.03, 0.04 mg / L ferulic acid, and 0.08, 0.1, 0.12 mg / L aspartic acid, and the callus induction rate was calculated, and the concentrations of other components were constant. As can be seen from Table 2, the combination of 2-3 mg / L 2, 4-D + 0.03 mg / L ferulic acid + 0.1 mg / L aspartic acid had the best effect in the induction of grade I and grade II spikelets. For grade III spikelets, the best combination was 3 mg / L 2, 4-D + 0.03 mg / L ferulic acid + 0.1 mg / L aspartic acid. Considering the induction rate of grade III (spikelet length above 25 mm), 3 mg / L 2, 4-D + 0.03 mg / L ferulic acid + 0.1 mg / L aspartic acid was the best concentration for spikelet axis callus induction.

[0035] Table 2 Effect of different callus induction medium on the induction rate of spikelet axis of green bamboo Spikelet length 2,4-D concentration (mg / L) Ferulic acid concentration (mg / L) Aspartic acid concentration (mg / L) Callus induction rate (%) Level Ⅰ (less than 15mm) 2 0.02 0.08 64.44±2.10 2 0.02 0.1 65.56±2.90 2 0.02 0.12 66.67±3.33 2 0.03 0.08 69.00±2.80 2 0.03 0.1 73.67±3.33 2 0.03 0.12 68.22±2.70 3 0.02 0.08 64.67±3.33 3 0.02 0.1 69.22±2.70 3 0.03 0.12 68.33±2.80 Level II (15-25 mm) 2 0.02 0.08 66.67±3.88 2 0.02 0.1 68.00±3.20 2 0.02 0.12 69.44±3.10 2 0.03 0.08 68.33±3.30 2 0.03 0.1 72.44±3.20 2 0.03 0.12 69.00±3.40 3 0.02 0.08 64.44±2.86 3 0.02 0.1 65.56±3.10 3 0.03 0.12 66.67±3.30 Level III (25 mm and above) 2 0.02 0.08 13.33±3.56 2 0.02 0.1 14.44±3.20 2 0.03 0.08 16.67±3.30 2 0.03 0.1 18.89±3.10 3 0.02 0.08 28.89±3.55 3 0.03 0.1 31.11±3.60 3 0.03 0.12 32.22±3.70 Example 3. This example provides a specific implementation method of a method for inducing plant regeneration based on spikelet axes of green bamboo.

[0036] The callus induced from the spikelet axis of green bamboo was inoculated into the medium prepared by orthogonal design, in which the concentration gradient of 6-BA was 2, 4, 6 mg / L, the concentration gradient of NAA was 0.1, 0.5, 1 mg / L, and the concentration gradient of TDZ was 0.1, 0.5, 1 mg / L, and the callus induction rate was counted. On the 5th day, one callus in group 5 turned green, and the number of green callus increased gradually with time. On the 10th day, the callus continued to differentiate, the green range increased, and one callus differentiated into adventitious roots. After 15 days, the number of green callus differentiation tended to be stable, and then gradually differentiated into bud points, and finally grew buds and leaves. In addition, 4 callus differentiated into flower buds and grew spikelets and anthers. The callus that grew spikelets was distributed in group 7 (1 case) and group 8 (3 cases). The results of the orthogonal experiment in Table 3 show that the effects of the three hormone types on callus differentiation rate are: 6-BA>NAA>TDZ, indicating that high concentrations of 6-BA are more likely to induce callus differentiation, but also more likely to promote callus browning and increase the possibility of callus flowering. In summary, the optimal hormone ratio combination formula for inducing callus differentiation of green bamboo spikelet axis is 4.0 mg / L 6-BA +0.5 mg / L NAA +0.5 mg / L TDZ.

[0037] Table 3 Orthogonal experimental design and result analysis of hormone ratios for callus differentiation of spikelet axis of green bamboo Group 6-BA (mg / L) NAA (mg / L) TDZ (mg / L) Callus differentiation rate （A） （B） （C） （%） 1 2 0.1 0.1 16.67 2 2 0.5 1 23.33 3 2 1 0.5 13.33 4 4 0.1 1 16.67 5 4 0.5 0.5 30.00 6 4 1 0.1 16.67 7 6 0.1 0.5 43.33 8 6 0.5 0.1 53.33 9 6 1 1 36.67 T1 53.33 76.67 86.67 T2 63.33 106.67 86.67 T3 133.33 66.67 76.67 T1 average 17.78 25.56 28.89 T2 average 21.11 35.56 28.89 T3 Average 44.44 22.22 25.56 R 26.67 13.33 3.33 In Table 3, Ti refers to the sum of the i-th level, Ti average refers to the average of the i-th level, and i = 1, 2, 3 R refers to the range.

[0038] In summary, the present invention establishes a callus induction system starting from the reproductive tissue of green-stalked bamboo, and successfully obtains regenerated plants, which provides a technical means for the rapid propagation of green-stalked bamboo seedlings.

[0039] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A method for inducing plant regeneration based on spikelet axis of green bamboo, characterized in that: include, Take the spikelets of flowering green bamboo, disinfect them, rinse them with sterile water, place them on sterile filter paper to absorb the water, and dissect to obtain the spikelet axis as explant; Inoculating the spikelet axis into a callus induction medium, culturing for 1-2 weeks in the dark, and inducing callus formation, wherein the induction medium components include MS medium, 2,4-D, KT, ferulic acid, aspartic acid, Yb, an amino acid mixture and sucrose; inoculating the callus into a differentiation medium, culturing under light conditions, and differentiating adventitious buds; The adventitious buds are transferred to a rooting medium, and after being cultured for a certain period of time, regenerated seedlings with relatively strong root systems are selected for seedling hardening, and transplanted to a suitable cultivation medium. After survival, they are placed in a greenhouse for growth. The rooting medium components include 1 / 2MS medium, IBA, NAA, ferric citrate, β-salicylic acid and sucrose.

2. The method for inducing plant regeneration based on spikelet axis of green bamboo as claimed in claim 1, characterized in that: The differentiation medium components include MS medium, 6-BA, NAA, hexanoic acid, adenine sulfate and sucrose.

3. The method for inducing plant regeneration based on spikelet axis of green bamboo as claimed in claim 1, characterized in that: The callus induction medium contains 2,4-D at a concentration of 2-5 mg / L, KT at a concentration of 0.2-1 mg / L, sucrose at a concentration of 20-30 g / L, ferulic acid at a concentration of 0.02-0.05 mg / L, aspartic acid at a concentration of 0.05-0.15 mg / L, Yb at a concentration of 3-5 μM, and an amino acid mixture at a concentration of 0.3-0.5 g / L.

4. The method for inducing plant regeneration based on spikelet axis of green bamboo as claimed in claim 1, characterized in that: The concentration of 6-BA in the differentiation medium is 4-8 mg / L, the concentration of NAA is 0.2-1 mg / L, the concentration of caproic acid is 0.1-0.5 mg / L, the concentration of adenine sulfate is 20-30 mg / L, and the concentration of sucrose is 20-30 g / L.

5. The method for inducing plant regeneration based on spikelet axis of green bamboo as claimed in claim 1, characterized in that: The explant of the spikelet axis is at least one of the base spikelet axis, the top spikelet axis or the middle spikelet axis of the flowering green bamboo.

6. The method for inducing plant regeneration based on spikelet axis of green bamboo as claimed in claim 1, characterized in that: The concentration of IBA in the rooting medium is 0.05-0.15 mg / L, the concentration of NAA is 0.5-1.5 mg / L, the concentration of sucrose is 15-25 g / L, the concentration of ferric citrate is 0.2-1 mg / L, and the concentration of beta-salicylic acid is 0.03-0.05 mg / L.

7. The method for inducing plant regeneration based on spikelet axis of green bamboo as claimed in claim 1, characterized in that: The induction culture step is carried out at a temperature of 24-28°C and the culture time is 1-2 weeks.

8. The method for inducing plant regeneration based on spikelet axis of green bamboo as claimed in claim 1, characterized in that: The rooting culture step of the regenerated seedlings is carried out at a temperature of 22-26° C. and the culture time is 4-6 weeks.

9. The method for inducing plant regeneration based on spikelet axis of green bamboo as claimed in claim 1, characterized in that: The induction medium components also include plant peptides and circular RNA.

10. The method for inducing plant regeneration based on spikelet axis of green bamboo as claimed in claim 1, characterized in that: During the treatment of the spikelet axis explants, the mercuric chloride disinfection time is 6-8 minutes, and the spikelet axis explants are rinsed with sterile water 5 times.

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