Composite culture medium and application thereof in rapid propagation of euphorbia colorata

Abstract

The invention discloses a composite culture medium and application thereof in rapid propagation of euphorbia colorata. The starting culture medium takes an MS culture medium as a basic culture medium and contains 6-benzyl purine, naphthylacetic acid, cane sugar and agar; the subculture medium takes an MS (Murashige and Skoog) culture medium as a basic culture medium, and contains 6-benzyl purine, methylthiodazine, thidiazuron, cane sugar and agar; the rooting culture medium takes a 1 / 2 MS culture medium as a basic culture medium, and contains indolebutyric acid and uniconazole, or diethyl aminoethyl hexanoate and isopentene adenine. By adopting the composite culture medium disclosed by the invention, the euphorbia colorata can be rapidly propagated. Meththiodazine and thidiazuron in the components of the culture medium are used in a combined mode, the propagation coefficient in the subculture stage is increased, synergistic interaction is achieved, and indolebutyric acid and uniconazole or diethyl aminoethyl hexanoate and isopenteneadenine are used in a combined mode, so that the number of roots and the average root length in the rooting stage are increased.

Description

Technical Field

[0001] The invention belongs to the technical field of tissue culture, and particularly relates to a composite culture medium and application thereof in the rapid propagation of colorful euphorbia. Background Art

[0002] Colored euphorbia, also known as Fei's euphorbia or Feilangkosi euphorbia, is a succulent plant belonging to the genus Euphorbia of the Euphorbiaceae family. The plant of colored euphorbia is low and clumps, with hypertrophic fleshy rhizomes. When the stem bark is broken, white milky slurry will flow out. Its leaves are diverse in shape, including round, long strips, diamonds, etc., with wrinkles or lace edges on the leaf edges, and rich leaf colors, including red, pink, white, light green, dark green and even close to black. Among the artificial variants, the colored leaf unicorn has yellow spots on the leaf surface. Colored euphorbia likes warm, dry and sunny environments, is drought-resistant, and is afraid of shade and waterlogging. Adequate light is required during planting, otherwise the plant is prone to leggy growth, affecting the ornamental value and health of the plant. In addition, the inflorescence of colored euphorbia is upright, a cup-shaped cyme, with larger bracts, and small flowers yellow-green or green.

[0003] In terms of medicinal value, it comes from the root of the plant. It is dug in spring before germination or in autumn when the stems and leaves wither. The residual stems and fibrous roots are removed, and it is washed and dried. It tastes bitter and pungent, is cold in nature, and has a slight toxicity. It enters the lung, spleen, and kidney meridians and has the effect of purging water and promoting bowel movements.

[0004] At present, the propagation of colorful euphorbia is mostly done by seed sowing and cuttings. When using seeds for propagation, its propagation cycle is long, the characteristics of the offspring are greatly separated, and the excellent characteristics of the mother plant cannot be maintained; when using cuttings for propagation, the incision secretes latex, the survival rate is low, which seriously affects the propagation coefficient of plants with excellent characteristics, and its growth potential in the later stage of cuttings is weak. That is, the immaturity of euphorbia cultivation and breeding technology has become an obstacle to the widespread application of euphorbia, and to a certain extent, it has limited the expansion of the planting range of euphorbia. Summary of the invention

[0005] The invention aims to provide a composite culture medium and application thereof in the rapid propagation of colorful euphorbia.

[0006] A composite culture medium comprises a starting culture medium, a subculture culture medium and a rooting culture medium. The starting culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, thioridazine, thidiazuron, sucrose and agar; and the rooting culture medium uses 1 / 2 MS culture medium as a basic culture medium and contains indolebutyric acid and uniconazole, or diethylaminoethyl hexanoate and isopentenyl adenine.

[0007] The concentration of 6-benzylpurine in the start-up culture medium is 0.5-1.5 mg / L; the concentration of naphthylacetic acid is 0.2-0.7 mg / L; the concentration of sucrose is 20-40 g / L; and the concentration of agar is 6-8 g / L.

[0008] The concentration of 6-benzylpurine in the subculture medium is 0.5-1.5 mg / L; the concentration of thioridazine is 0.1-0.3 mg / L; the concentration of thidiazuron is 0.02-0.1 mg / L; the concentration of sucrose is 20-40 g / L; and the concentration of agar is 6-8 g / L.

[0009] The concentration of indolebutyric acid in the rooting medium is 0.5-1.5 mg / L; the concentration of uniconazole is 0.5-1.5 mg / L.

[0010] The concentration of diethylaminoethyl hexanoate in the rooting medium is 1.5-4.5 mg / L; the concentration of isopentenyl adenine is 0.05-0.5 mg / L.

[0011] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0012] (1) Preparing explants: taking young stem segments and stem tips of Euphorbia pulex, washing, disinfecting, and cutting them in sequence to obtain explants;

[0013] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 20-22 days, and when the lateral buds germinate and grow to 2-3 cm, they are transferred to subculture;

[0014] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culture for 25-30 days to obtain a single plant;

[0015] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0016] (5) Transplantation: The rooted seedlings were moved to a greenhouse for acclimatization for 3-4 days, and then hardened for 2-3 days before transplanting.

[0017] In step (1), young stem segments and stem tips of Euphorbia pulegiae are taken and leaves are cut off, placed in a container, detergent is added, and rinsed with water for 30-35 minutes, then disinfected with 75% alcohol for 20-40 seconds under sterile conditions, then soaked in 0.1% mercuric chloride solution for 2-4 minutes, finally rinsed with sterile water for 3-5 times, and cut into 0.5-1.0 cm stem tips and stem segments with axillary buds, and the surface moisture is absorbed with sterile filter paper to obtain explants.

[0018] The transplanting in step (5) comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and sterilizing in sequence to obtain a sterilized matrix; sterilizing the roots of the rooting seedlings, then dipping them in rooting liquid and transplanting them into the sterilized matrix; controlling the temperature at the initial stage of transplanting to 18-28°C, the relative humidity to 70-80%, and the refractive index to 50-60%; applying a 10-20 ppm amino acid water-soluble fertilizer GGR solution once every other week and managing the growth for 30-40 days.

[0019] The transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0020] The transplanting medium is disinfected by using a thiophanate-methyl solution diluted 800 times; the roots of the rooted seedlings are disinfected by using a thiophanate-methyl solution.

[0021] The invention has the beneficial effects that the composite culture medium of the invention can be used to rapidly propagate colorful euphorbia. The combination of thioridazine and thidiazuron in the culture medium components improves the propagation coefficient in the subculture stage, and synergistically enhances the effect. The combination of indolebutyric acid and uniconazole, or ampicillin and isopentenyl adenine, increases the number of roots and the average root length in the rooting stage. DETAILED DESCRIPTION

[0022] In order to facilitate the understanding of the present invention, the present invention will be described more fully below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0023] Example 1

[0024] A composite culture medium comprises a start-up culture medium, a subculture culture medium and a rooting culture medium, wherein the start-up culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, thioridazine, thidiazuron, sucrose and agar; the rooting culture medium uses a 1 / 2MS culture medium as a basic culture medium and contains indolebutyric acid and uniconazole. The concentration of 6-benzylpurine in the start-up culture medium is 1 mg / L; the concentration of naphthylacetic acid is 0.5 mg / L; the concentration of sucrose is 30 g / L; and the concentration of agar is 7 g / L. The concentration of 6-benzylpurine in the subculture culture medium is 1 mg / L; the concentration of thioridazine is 0.2 mg / L; the concentration of thidiazuron is 0.05 mg / L; the concentration of sucrose is 30 g / L; and the concentration of agar is 7 g / L. The concentration of indolebutyric acid in the rooting medium is 1 mg / L; the concentration of uniconazole is 1 mg / L.

[0025] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0026] (1) Explant preparation: Take young stem segments and stem tips of Euphorbia pulegosa and cut off the leaves, put them in a container, add detergent and rinse with water for 32 minutes, then sterilize with 75% alcohol for 30 seconds under sterile conditions, soak in 0.1% mercuric chloride solution for 3 minutes, and finally rinse with sterile water 4 times, cut into 0.8 cm stem tips and stem segments with axillary buds, and dry the surface moisture with sterile filter paper to obtain explants;

[0027] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 21 days, and when the lateral buds germinate and grow to 2.5 cm, they are transferred to subculture;

[0028] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culturing for 28 days to obtain a single plant;

[0029] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0030] (5) Transplantation and cultivation: the rooting seedlings are moved to a greenhouse for acclimatization for 3 days, and then transplanted after hardening for 2 days. The transplanting comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and disinfecting (using 800-fold diluted thiophanate-methyl solution) in sequence to obtain a disinfected matrix; disinfecting the roots of the rooting seedlings (using thiophanate-methyl solution), then dipping them in rooting liquid and transplanting them into the disinfected matrix; at the initial stage of transplanting, the temperature is controlled at 23°C, the relative humidity is 75%, and the refractive index is 55%; applying 15 ppm amino acid water-soluble fertilizer GGR solution once every week and managing the growth for 35 days; the transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0031] Example 2

[0032] A composite culture medium comprises a start-up culture medium, a subculture culture medium and a rooting culture medium, wherein the start-up culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, thioridazine, thidiazuron, sucrose and agar; the rooting culture medium uses a 1 / 2MS culture medium as a basic culture medium and contains aminoethyl hexanoate and isopentenyl adenine. The concentration of 6-benzylpurine in the start-up culture medium is 0.8 mg / L; the concentration of naphthylacetic acid is 0.3 mg / L; the concentration of sucrose is 25 g / L; and the concentration of agar is 6 g / L. The concentration of 6-benzylpurine in the subculture culture medium is 0.8 mg / L; the concentration of thioridazine is 0.1 mg / L; the concentration of thidiazuron is 0.03 mg / L; the concentration of sucrose is 25 g / L; and the concentration of agar is 6 g / L. The concentration of diethylaminoethyl hexanoate in the rooting medium is 2 mg / L; the concentration of isopentenyl adenine is 0.2 mg / L.

[0033] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0034] (1) Explant preparation: Take young stem segments and stem tips of Euphorbia pulegosa and cut off the leaves, put them in a container, add detergent and rinse with water for 30 min, then sterilize with 75% alcohol for 25 s under sterile conditions, soak in 0.1% mercuric chloride solution for 2 min, and finally rinse with sterile water three times, cut into 0.5 cm stem tips and stem segments with axillary buds, and absorb the surface moisture with sterile filter paper to obtain explants;

[0035] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 20 days, and when the lateral buds germinate and grow to 2 cm, they are transferred to subculture;

[0036] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culture for 25 days to obtain a single plant;

[0037] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0038] (5) Transplantation and cultivation: the rooting seedlings are moved to a greenhouse for acclimatization for 3 days, and then transplanted after hardening for 2 days. The transplanting comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and disinfecting (using 800-fold diluted thiophanate-methyl solution) in sequence to obtain a disinfected matrix; disinfecting the roots of the rooting seedlings (using thiophanate-methyl solution), then dipping them in rooting liquid and transplanting them into the disinfected matrix; at the initial stage of transplanting, the temperature is controlled at 20°C, the relative humidity is 70%, and the refractive index is 50%; applying 12 ppm amino acid water-soluble fertilizer GGR solution once every week and managing the growth for 30 days; the transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0039] Example 3

[0040] A composite culture medium comprises a start-up culture medium, a subculture culture medium and a rooting culture medium, wherein the start-up culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, thioridazine, thidiazuron, sucrose and agar; the rooting culture medium uses a 1 / 2MS culture medium as a basic culture medium and contains indolebutyric acid and uniconazole. The concentration of 6-benzylpurine in the start-up culture medium is 1.5 mg / L; the concentration of naphthylacetic acid is 0.6 mg / L; the concentration of sucrose is 40 g / L; and the concentration of agar is 8 g / L. The concentration of 6-benzylpurine in the subculture culture medium is 1.5 mg / L; the concentration of thioridazine is 0.3 mg / L; the concentration of thidiazuron is 0.08 mg / L; the concentration of sucrose is 40 g / L; and the concentration of agar is 8 g / L. The concentration of indolebutyric acid in the rooting medium is 1 mg / L; the concentration of uniconazole is 1 mg / L.

[0041] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0042] (1) Explant preparation: Take young stem segments and stem tips of Euphorbia pulegosa and cut off the leaves, put them in a container, add detergent and rinse with water for 35 minutes, then sterilize with 75% alcohol for 40 seconds under sterile conditions, soak in 0.1% mercuric chloride solution for 4 minutes, and finally rinse with sterile water 5 times, cut into 1.0 cm stem tips and stem segments with axillary buds, and absorb the surface moisture with sterile filter paper to obtain explants;

[0043] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 22 days, and when the lateral buds germinate and grow to 3 cm, they are transferred to subculture;

[0044] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culture for 30 days to obtain a single plant;

[0045] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0046] (5) Transplantation and cultivation: the rooting seedlings are moved to a greenhouse for acclimatization for 4 days, and then transplanted after hardening for 3 days. The transplanting comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and disinfecting (using 800-fold diluted thiophanate-methyl solution) in sequence to obtain a disinfected matrix; disinfecting the roots of the rooting seedlings (using thiophanate-methyl solution), then dipping them in rooting liquid and transplanting them into the disinfected matrix; at the initial stage of transplanting, the temperature is controlled at 28°C, the relative humidity is 80%, and the refractive index is 60%; applying 20 ppm amino acid water-soluble fertilizer GGR solution once every week and managing the growth for 40 days; the transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0047] Comparative Example 1

[0048] A composite culture medium comprises a start-up culture medium, a subculture culture medium and a rooting culture medium, wherein the start-up culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, thioridazine, sucrose and agar; the rooting culture medium uses a 1 / 2MS culture medium as a basic culture medium and contains indolebutyric acid and uniconazole. The concentration of 6-benzylpurine in the start-up culture medium is 1 mg / L; the concentration of naphthylacetic acid is 0.5 mg / L; the concentration of sucrose is 30 g / L; and the concentration of agar is 7 g / L. The concentration of 6-benzylpurine in the subculture culture medium is 1 mg / L; the concentration of thioridazine is 0.25 mg / L; the concentration of sucrose is 30 g / L; and the concentration of agar is 7 g / L. The concentration of indolebutyric acid in the rooting culture medium is 1 mg / L; and the concentration of uniconazole is 1 mg / L.

[0049] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0050] (1) Explant preparation: Take young stem segments and stem tips of Euphorbia pulegosa and cut off the leaves, put them in a container, add detergent and rinse with water for 32 minutes, then sterilize with 75% alcohol for 30 seconds under sterile conditions, soak in 0.1% mercuric chloride solution for 3 minutes, and finally rinse with sterile water 4 times, cut into 0.8 cm stem tips and stem segments with axillary buds, and dry the surface moisture with sterile filter paper to obtain explants;

[0051] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 21 days, and when the lateral buds germinate and grow to 2.5 cm, they are transferred to subculture;

[0052] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culturing for 28 days to obtain a single plant;

[0053] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0054] (5) Transplantation and cultivation: the rooting seedlings are moved to a greenhouse for acclimatization for 3 days, and then transplanted after hardening for 2 days. The transplanting comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and disinfecting (using 800-fold diluted thiophanate-methyl solution) in sequence to obtain a disinfected matrix; disinfecting the roots of the rooting seedlings (using thiophanate-methyl solution), then dipping them in rooting liquid and transplanting them into the disinfected matrix; at the initial stage of transplanting, the temperature is controlled at 23°C, the relative humidity is 75%, and the refractive index is 55%; applying 15 ppm amino acid water-soluble fertilizer GGR solution once every week and managing the growth for 35 days; the transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0055] Comparative Example 2

[0056] A composite culture medium comprises a start-up culture medium, a subculture culture medium and a rooting culture medium, wherein the start-up culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, thidiazuron, sucrose and agar; the rooting culture medium uses a 1 / 2MS culture medium as a basic culture medium and contains indolebutyric acid and uniconazole. The concentration of 6-benzylpurine in the start-up culture medium is 1 mg / L; the concentration of naphthylacetic acid is 0.5 mg / L; the concentration of sucrose is 30 g / L; and the concentration of agar is 7 g / L. The concentration of 6-benzylpurine in the subculture culture medium is 1 mg / L; the concentration of thidiazuron is 0.25 mg / L; the concentration of sucrose is 30 g / L; and the concentration of agar is 7 g / L. The concentration of indolebutyric acid in the rooting culture medium is 1 mg / L; and the concentration of uniconazole is 1 mg / L.

[0057] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0058] (1) Explant preparation: Take young stem segments and stem tips of Euphorbia pulegosa and cut off the leaves, put them in a container, add detergent and rinse with water for 32 minutes, then sterilize with 75% alcohol for 30 seconds under sterile conditions, soak in 0.1% mercuric chloride solution for 3 minutes, and finally rinse with sterile water 4 times, cut into 0.8 cm stem tips and stem segments with axillary buds, and dry the surface moisture with sterile filter paper to obtain explants;

[0059] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 21 days, and when the lateral buds germinate and grow to 2.5 cm, they are transferred to subculture;

[0060] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culturing for 28 days to obtain a single plant;

[0061] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0062] (5) Transplantation and cultivation: the rooting seedlings are moved to a greenhouse for acclimatization for 3 days, and then transplanted after hardening for 2 days. The transplanting comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and disinfecting (using 800-fold diluted thiophanate-methyl solution) in sequence to obtain a disinfected matrix; disinfecting the roots of the rooting seedlings (using thiophanate-methyl solution), then dipping them in rooting liquid and transplanting them into the disinfected matrix; at the initial stage of transplanting, the temperature is controlled at 23°C, the relative humidity is 75%, and the refractive index is 55%; applying 15 ppm amino acid water-soluble fertilizer GGR solution once every week and managing the growth for 35 days; the transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0063] Comparative Example 3

[0064] A composite culture medium comprises a start-up culture medium, a subculture culture medium and a rooting culture medium, wherein the start-up culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, thioridazine, thidiazuron, sucrose and agar; the rooting culture medium uses a 1 / 2MS culture medium as a basic culture medium and contains diethylaminoethyl. The concentration of 6-benzylpurine in the start-up culture medium is 0.8 mg / L; the concentration of naphthylacetic acid is 0.3 mg / L; the concentration of sucrose is 25 g / L; and the concentration of agar is 6 g / L. The concentration of 6-benzylpurine in the subculture culture medium is 0.8 mg / L; the concentration of thioridazine is 0.1 mg / L; the concentration of thidiazuron is 0.03 mg / L; the concentration of sucrose is 25 g / L; and the concentration of agar is 6 g / L. The concentration of diethylaminoethyl hexanoate in the rooting medium is 2.2 mg / L.

[0065] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0066] (1) Explant preparation: Take young stem segments and stem tips of Euphorbia pulegosa and cut off the leaves, put them in a container, add detergent and rinse with water for 30 min, then sterilize with 75% alcohol for 25 s under sterile conditions, soak in 0.1% mercuric chloride solution for 2 min, and finally rinse with sterile water three times, cut into 0.5 cm stem tips and stem segments with axillary buds, and absorb the surface moisture with sterile filter paper to obtain explants;

[0067] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 20 days, and when the lateral buds germinate and grow to 2 cm, they are transferred to subculture;

[0068] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culture for 25 days to obtain a single plant;

[0069] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0070] (5) Transplantation and cultivation: the rooting seedlings are moved to a greenhouse for acclimatization for 3 days, and then transplanted after hardening for 2 days. The transplanting comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and disinfecting (using 800-fold diluted thiophanate-methyl solution) in sequence to obtain a disinfected matrix; disinfecting the roots of the rooting seedlings (using thiophanate-methyl solution), then dipping them in rooting liquid and transplanting them into the disinfected matrix; at the initial stage of transplanting, the temperature is controlled at 20°C, the relative humidity is 70%, and the refractive index is 50%; applying 12 ppm amino acid water-soluble fertilizer GGR solution once every week and managing the growth for 30 days; the transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0071] Comparative Example 4

[0072] A composite culture medium comprises a start-up culture medium, a subculture culture medium and a rooting culture medium, wherein the start-up culture medium uses an MS culture medium as a basic culture medium and contains 6-benzyladenine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzyladenine, thioridazine, thidiazuron, sucrose and agar; the rooting culture medium uses a 1 / 2MS culture medium as a basic culture medium and contains isopentenyladenine. The concentration of 6-benzyladenine in the start-up culture medium is 0.8 mg / L; the concentration of naphthylacetic acid is 0.3 mg / L; the concentration of sucrose is 25 g / L; and the concentration of agar is 6 g / L. The concentration of 6-benzyladenine in the subculture culture medium is 0.8 mg / L; the concentration of thioridazine is 0.1 mg / L; the concentration of thidiazuron is 0.03 mg / L; the concentration of sucrose is 25 g / L; and the concentration of agar is 6 g / L. The concentration of isopentenyl adenine in the rooting medium is 2.2 mg / L.

[0073] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0074] (1) Explant preparation: Take young stem segments and stem tips of Euphorbia pulegosa and cut off the leaves, put them in a container, add detergent and rinse with water for 30 min, then sterilize with 75% alcohol for 25 s under sterile conditions, soak in 0.1% mercuric chloride solution for 2 min, and finally rinse with sterile water three times, cut into 0.5 cm stem tips and stem segments with axillary buds, and absorb the surface moisture with sterile filter paper to obtain explants;

[0075] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 20 days, and when the lateral buds germinate and grow to 2 cm, they are transferred to subculture;

[0076] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culture for 25 days to obtain a single plant;

[0077] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0078] (5) Transplantation and cultivation: the rooting seedlings are moved to a greenhouse for acclimatization for 3 days, and then transplanted after hardening for 2 days. The transplanting comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and disinfecting (using 800-fold diluted thiophanate-methyl solution) in sequence to obtain a disinfected matrix; disinfecting the roots of the rooting seedlings (using thiophanate-methyl solution), then dipping them in rooting liquid and transplanting them into the disinfected matrix; at the initial stage of transplanting, the temperature is controlled at 20°C, the relative humidity is 70%, and the refractive index is 50%; applying 12 ppm amino acid water-soluble fertilizer GGR solution once every week and managing the growth for 30 days; the transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0079] Comparative Example 5

[0080] A composite culture medium comprises a start-up culture medium, a subculture culture medium and a rooting culture medium, wherein the start-up culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, thioridazine, thidiazuron, sucrose and agar; the rooting culture medium uses a 1 / 2MS culture medium as a basic culture medium and contains indolebutyric acid. The concentration of 6-benzylpurine in the start-up culture medium is 1.5 mg / L; the concentration of naphthylacetic acid is 0.6 mg / L; the concentration of sucrose is 40 g / L; and the concentration of agar is 8 g / L. The concentration of 6-benzylpurine in the subculture culture medium is 1.5 mg / L; the concentration of thioridazine is 0.3 mg / L; the concentration of thidiazuron is 0.08 mg / L; the concentration of sucrose is 40 g / L; and the concentration of agar is 8 g / L. The concentration of indolebutyric acid in the rooting medium is 2 mg / L.

[0081] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0082] (1) Explant preparation: Take young stem segments and stem tips of Euphorbia pulegosa and cut off the leaves, put them in a container, add detergent and rinse with water for 35 minutes, then sterilize with 75% alcohol for 40 seconds under sterile conditions, soak in 0.1% mercuric chloride solution for 4 minutes, and finally rinse with sterile water 5 times, cut into 1.0 cm stem tips and stem segments with axillary buds, and absorb the surface moisture with sterile filter paper to obtain explants;

[0083] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 22 days, and when the lateral buds germinate and grow to 3 cm, they are transferred to subculture;

[0084] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culture for 30 days to obtain a single plant;

[0085] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0086] (5) Transplantation and cultivation: the rooting seedlings are moved to a greenhouse for acclimatization for 4 days, and then transplanted after hardening for 3 days. The transplanting comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and disinfecting (using 800-fold diluted thiophanate-methyl solution) in sequence to obtain a disinfected matrix; disinfecting the roots of the rooting seedlings (using thiophanate-methyl solution), then dipping them in rooting liquid and transplanting them into the disinfected matrix; at the initial stage of transplanting, the temperature is controlled at 28°C, the relative humidity is 80%, and the refractive index is 60%; applying 20 ppm amino acid water-soluble fertilizer GGR solution once every week and managing the growth for 40 days; the transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0087] Comparative Example 6

[0088] A composite culture medium comprises a start-up culture medium, a subculture culture medium and a rooting culture medium, wherein the start-up culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses an MS culture medium as a basic culture medium and contains 6-benzylpurine, thioridazine, thidiazuron, sucrose and agar; the rooting culture medium uses a 1 / 2MS culture medium as a basic culture medium and contains uniconazole. The concentration of 6-benzylpurine in the start-up culture medium is 1.5 mg / L; the concentration of naphthylacetic acid is 0.6 mg / L; the concentration of sucrose is 40 g / L; and the concentration of agar is 8 g / L. The concentration of 6-benzylpurine in the subculture culture medium is 1.5 mg / L; the concentration of thioridazine is 0.3 mg / L; the concentration of thidiazuron is 0.08 mg / L; the concentration of sucrose is 40 g / L; and the concentration of agar is 8 g / L. The concentration of uniconazole in the rooting medium is 2 mg / L.

[0089] A rapid propagation method for colorful euphorbia is carried out according to the following steps:

[0090] (1) Explant preparation: Take young stem segments and stem tips of Euphorbia pulegosa and cut off the leaves, put them in a container, add detergent and rinse with water for 35 minutes, then sterilize with 75% alcohol for 40 seconds under sterile conditions, soak in 0.1% mercuric chloride solution for 4 minutes, and finally rinse with sterile water 5 times, cut into 1.0 cm stem tips and stem segments with axillary buds, and absorb the surface moisture with sterile filter paper to obtain explants;

[0091] (2) Initiation culture: The explants are placed in the initiation culture medium and cultured for 22 days, and when the lateral buds germinate and grow to 3 cm, they are transferred to subculture;

[0092] (3) Subculture: the germinated lateral buds are cut off and transferred to the subculture medium for culture for 30 days to obtain a single plant;

[0093] (4) Rooting culture: transferring the single plant into the rooting medium for culture to obtain rooted seedlings;

[0094] (5) Transplantation and cultivation: the rooting seedlings are moved to a greenhouse for acclimatization for 4 days, and then transplanted after hardening for 3 days. The transplanting comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and disinfecting (using 800-fold diluted thiophanate-methyl solution) in sequence to obtain a disinfected matrix; disinfecting the roots of the rooting seedlings (using thiophanate-methyl solution), then dipping them in rooting liquid and transplanting them into the disinfected matrix; at the initial stage of transplanting, the temperature is controlled at 28°C, the relative humidity is 80%, and the refractive index is 60%; applying 20 ppm amino acid water-soluble fertilizer GGR solution once every week and managing the growth for 40 days; the transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

[0095] Experimental Example 1:

[0096] The composite culture medium of Example 1, Comparative Example 1 and Comparative Example 2 was used as the experimental object. According to the rapid propagation method, samples were taken at the subculture stage to determine the propagation coefficient. Each group of experiments was measured 5 times and the average value was taken. SPSS24.0 software was used for statistical analysis. The quantitative data results were expressed as (mean ± standard deviation), the Kolmogorov-Smirnov test was used for data normality test, and the t-test was used to compare the mean differences between the two groups for data that met the normal distribution, with P < 0.05 as the difference being statistically significant. The results are shown in Table 1:

[0097] Table 1

[0098]

[0099] Note: * represents P < 0.05 compared with the group in Example 1.

[0100] Experimental Example 2:

[0101] The composite culture medium of Example 2, Comparative Example 3 and Comparative Example 4 was used as the experimental object. According to the rapid propagation method, samples were taken in the rooting culture stage to measure the average root length and the number of roots. Each group of experiments was measured 5 times and the average value was taken. The measurement results are shown in Table 2:

[0102] Table 2

[0103]

[0104] Note: * represents P < 0.05 compared with the group in Example 2.

[0105] The composite culture medium of Example 3, Comparative Example 5 and Comparative Example 6 was used as the experimental object. According to the rapid propagation method, samples were taken in the rooting culture stage to measure the average root length and the number of roots. Each group of experiments was measured 5 times and the average value was taken. The measurement results are shown in Table 3:

[0106] Table 3

[0107]

[0108] Note: * represents P < 0.05 compared with the group in Example 3.

[0109] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A composite culture medium comprising a starting culture medium, a subculture culture medium and a rooting culture medium, characterized in that: The initiation culture medium uses MS culture medium as the basic culture medium and contains 6-benzylpurine, naphthylacetic acid, sucrose and agar; the subculture culture medium uses MS culture medium as the basic culture medium and contains 6-benzylpurine, thioridazine, thidiazuron, sucrose and agar; the rooting culture medium uses 1 / 2MS culture medium as the basic culture medium and contains indolebutyric acid and uniconazole, or aminoethyl hexanoate and isopentenyl adenine.

2. The composite culture medium according to claim 1, characterized in that The concentration of 6-benzylpurine in the start-up culture medium is 0.5-1.5 mg / L; the concentration of naphthylacetic acid is 0.2-0.7 mg / L; the concentration of sucrose is 20-40 g / L; and the concentration of agar is 6-8 g / L.

3. The composite culture medium according to claim 1, characterized in that The concentration of 6-benzylpurine in the subculture medium is 0.5-1.5 mg / L; the concentration of thioridazine is 0.1-0.3 mg / L; the concentration of thidiazuron is 0.02-0.1 mg / L; the concentration of sucrose is 20-40 g / L; and the concentration of agar is 6-8 g / L.

4. The composite culture medium according to claim 1, characterized in that The concentration of indolebutyric acid in the rooting medium is 0.5-1.5 mg / L; the concentration of uniconazole is 0.5-1.5 mg / L.

5. The composite culture medium according to claim 1, characterized in that The concentration of ammonium hexanoate in the rooting medium is 1.5-4.5 mg / L; the concentration of isopentenyl adenine is 0.05-0.5 mg / L.

6. A method for rapid propagation of colorful euphorbia, characterized in that: Follow these steps: (1) Preparing explants: taking young stem segments and stem tips of Euphorbia pulex, washing, disinfecting, and cutting them in sequence to obtain explants; (2) Initiation culture: the explant is inoculated into the initiation culture medium of claim 1 and cultured for 20-22 days, and when the lateral buds germinate and grow to 2-3 cm, they are transferred to subculture; (3) Subculture: cutting off the germinated lateral buds and transferring them to the subculture medium of claim 1 for culturing for 25-30 days to obtain a single plant; (4) Rooting culture: transferring the single plant into the rooting medium of claim 1 for culturing to obtain rooted seedlings; (5) Transplantation: The rooted seedlings were moved to a greenhouse for acclimatization for 3-4 days, and then hardened for 2-3 days before transplanting.

7. The rapid propagation method of colorful euphorbia according to claim 6, characterized in that: In step (1), young stem segments and stem tips of Euphorbia pulegiae are taken and leaves are cut off, placed in a container, detergent is added, and rinsed with water for 30-35 minutes, then disinfected with 75% alcohol for 20-40 seconds under sterile conditions, then soaked in 0.1% mercuric chloride solution for 2-4 minutes, finally rinsed with sterile water for 3-5 times, and cut into 0.5-1.0 cm stem tips and stem segments with axillary buds, and the surface moisture is absorbed with sterile filter paper to obtain explants.

8. The rapid propagation method of colorful euphorbia according to claim 6, characterized in that: The transplanting in step (5) comprises the following steps: exposing the transplanting matrix to sunlight, mixing, stirring and sterilizing in sequence to obtain a sterilized matrix; sterilizing the roots of the rooting seedlings, then dipping them in rooting liquid and transplanting them into the sterilized matrix; controlling the temperature at the initial stage of transplanting to 18-28°C, the relative humidity to 70-80%, and the refractive index to 50-60%; applying a 10-20 ppm amino acid water-soluble fertilizer GGR solution once every other week and managing the growth for 30-40 days.

9. The rapid propagation method of colorful euphorbia according to claim 8, characterized in that: The transplanting matrix comprises the following raw materials in parts by weight: 7 parts of peat, 2 parts of perlite and 1 part of river sand.

10. The rapid propagation method of colorful euphorbia according to claim 8, characterized in that: The transplanting medium is disinfected by using a thiophanate-methyl solution diluted 800 times; the roots of the rooted seedlings are disinfected by using a thiophanate-methyl solution.

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