Open type tissue culture method and application of dionaea muscipula

Abstract

The invention relates to the technical field of plant culture, and particularly discloses an open type tissue culture method and application of dionaea muscipula. Comprising the following steps: (1) selecting leaves of dionaea muscipula, cleaning and disinfecting; (2) preparing a liquid culture medium A; (3) adding a liquid culture medium A and the disinfected dionaea muscipula leaves into a container, sealing, and culturing under red light to obtain dionaea muscipula leaves with dense adventitious buds; (4) preparing a liquid culture medium B; (5) replacing the culture medium with a liquid culture medium B in the container, adding the dionaea muscipula leaves with dense adventitious buds, sealing, and culturing under the common illumination of red light and blue light to obtain the dionaea muscipula leaves with roots; and (6) taking out the leaves of the dionaea muscipula with roots, cleaning, planting in a matrix, and performing seedling hardening culture under natural light to complete open tissue culture of the dionaea muscipula. The open type tissue culture method of the dionaea muscipula provided by the invention achieves the effects of shorter tissue culture time and more efficient breeding.

Description

Technical Field

[0001] The invention relates to the technical field of plant cultivation, and in particular to an open tissue culture method and application of Venus flytrap. Background Art

[0002] The Venus flytrap is an emerging ornamental plant with unique ornamental value and is loved by the market. The Venus flytrap belongs to the Droseraceae family and the Venus flytrap genus. The Venus flytrap is a vascular plant and a very popular insectivorous plant with complete roots, stems, leaves, flowers and seeds. Its leaves are the most important and obvious part, which has the function of preying on insects. The Venus flytrap is 10 to 30 cm tall; the stem is upright and slender; the basal leaves are small and round, and wither when the flowers bloom; the stem leaves are alternate, with thin petioles, about 1 cm long, crescent-shaped or fan-shaped, with a concave base, divided into two halves, and the edges and leaf surfaces are covered with many fine hairs, secreting mucus, in the shape of dewdrops, and there are 3 roots in the middle of the leaf margin that are sensitive to stimuli. The leaves usually open outward, and the nectaries on the leaf margins emit a sweet smell. Scape 30 cm high; racemose inflorescence; florets white; 5 sepals, united at base, ovate, with irregular marginal teeth, glandular hairs on margins; 5 petals, white, narrowly obovate, longer than sepals, with colored longitudinal stripes; 5 stamens, slender filaments; 1 pistil, superior ovary, 1-chambered; 3 styles, finger-like 4-lobed; capsule; seeds small, elliptic reticulate, with longitudinal stripes.

[0004] Although Venus flytraps have a high market demand, conventional tissue culture currently has problems such as high environmental requirements and long cycles. In order to shorten the cultivation cycle, reduce cultivation costs, and improve economic benefits, there is a strong demand for new rapid propagation technologies. Therefore, it is necessary to provide an open tissue culture method and application of Venus flytraps to quickly breed Venus flytraps and achieve a shorter and more efficient proliferation effect than conventional tissue culture. Summary of the invention

[0005] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the present invention provides an open tissue culture method and application of Venus flytrap to quickly breed Venus flytrap and achieve a shorter and more efficient proliferation effect than conventional tissue culture.

[0006] A first aspect of the present invention provides an open tissue culture method for Venus flytrap.

[0007] Specifically, the following steps are included:

[0008] (1) selecting a leaf of Venus flytrap, washing it, and soaking it in 0.1-0.2% chlorobromoisocyanuric acid to disinfect it, thereby obtaining a disinfected leaf of Venus flytrap;

[0009] (2) Add water to a 1.5-2 g / L solution of sugar-free and agar-free MS medium, boil it, add 0.1-3 g / L sucrose, 0.01-1 g / L chlorobromoisocyanuric acid, 0.2-0.5 g / L potassium rosinate, 0.05-0.1 g / L potassium acetate, 0.1-0.2 g / L salicylic acid, 0.1-0.2 g / L glutaraldehyde, and 1.5-3 mg / L cytokinin, and stir to prepare a liquid medium A;

[0010] (3) adding the liquid culture medium A described in step (2) into a container to form a liquid layer with a thickness of 2 to 3 mm, adding the disinfected Venus flytrap leaves described in step (1), sealing, and culturing under 950 to 1050 lux red light for 10 to 15 days to obtain Venus flytrap leaves with dense adventitious buds;

[0011] (4) Add water to the sugar-free and agar-free MS medium to make a 1.5-2 g / L solution, boil it, add 0.2-0.3 g / L sucrose, 0.01-1 g / L chlorobromoisocyanuric acid, 0.2-0.5 g / L potassium rosinate, 0.05-0.1 g / L potassium acetate, 0.1-0.2 g / L salicylic acid, 0.1-0.2 g / L glutaraldehyde, and 0.5-1 mg / L indolebutyric acid, and stir to prepare a liquid medium B;

[0012] (5) replacing the culture medium in the container with the liquid culture medium B described in step (4) to form a liquid layer with a thickness of 2 to 3 mm, adding the Venus flytrap leaves with dense adventitious buds described in step (3), sealing, and culturing for 10 to 15 days under the co-illumination of 950 to 1050 lux red light and 1950 to 2050 lux blue light to obtain Venus flytrap leaves with roots;

[0013] (6) Take out the leaf of the Venus flytrap with roots described in step (5), clean it, and plant it in a substrate, and cultivate it under natural light for 4 to 6 days to complete the open tissue culture of the Venus flytrap.

[0014] Preferably, in step (1), the Venus flytrap leaf is a healthy Venus flytrap leaf with buds.

[0015] Preferably, in step (1), the immersion disinfection time is 20 to 40 minutes.

[0016] Further preferably, in step (1), the immersion disinfection time is 25 to 35 minutes.

[0017] More preferably, in step (1), the immersion disinfection time is 30 minutes.

[0018] Preferably, in step (2), the cytokinin comprises at least one of indolebutyric acid.

[0019] Preferably, in step (3), the wavelength of the red light is 620-720 nm.

[0020] Preferably, in step (3), the illumination intensity of the red light is 980 to 1020 lux.

[0021] Further preferably, in step (3), the illumination intensity of the red light is 1000 lux.

[0022] Preferably, it is characterized in that, in step (5), the wavelength of the blue light is 400-520 nm.

[0023] Preferably, in step (5), the illumination intensity of the blue light is 1980-2020 lux.

[0024] Further preferably, in step (5), the illumination intensity of the blue light is 2000 lux.

[0025] Preferably, in step (6), the culturing time is 4 to 5 days.

[0026] Further preferably, in step (6), the culturing time is 5 days.

[0027] Preferably, in step (6), the substrate is sphagnum moss.

[0028] Preferably, in step (6), the illumination intensity of the natural light is 2000-2500 lux.

[0029] Further preferably, in step (6), the illumination intensity of the natural light is 2000-2300 lux.

[0030] More preferably, in step (6), the illumination intensity of the natural light is 2000 lux.

[0031] Preferably, in step (6), the humidity of the seedling training culture is 70-90%.

[0032] A second aspect of the present invention provides an application of an open tissue culture method for Venus flytraps in the rapid propagation of Venus flytraps.

[0033] Specifically, the open tissue culture method of Venus flytrap is provided by the first aspect of the present invention.

[0034] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0035] The open tissue culture method of Venus flytrap provided by the present invention omits the dedifferentiation step and uses hormone catalysis and heterotrophic conditions to quickly breed Venus flytrap leaves with buds, thereby achieving the effect of shorter tissue culture time and more efficient breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 This is a picture of the Venus flytrap mother plant selected during tissue culture in Example 1 of the present invention;

[0037] Figure 2 This is a picture of the Venus flytrap seedlings cultured in Example 1 of the present invention. DETAILED DESCRIPTION

[0038] In order to make the technical scheme of the present invention more clearly understood by those skilled in the art, the following embodiments are listed for illustration. It should be pointed out that the following embodiments do not limit the protection scope of the present invention.

[0039] Unless otherwise specified, the raw materials, reagents or devices used in the following examples can be obtained from conventional commercial sources or by existing known methods.

[0040] Example 1

[0041] The invention discloses an open tissue culture method for Venus flytrap.

[0042] The following steps are involved:

[0043] (1) Select a healthy Venus flytrap mother (such as Figure 1 As shown), select healthy Venus flytrap leaves with buds, wash with distilled water, and use 0.1% chlorobromoisocyanuric acid to immerse and disinfect for 30 minutes to obtain disinfected Venus flytrap leaves, and then wash and disinfect the tissue culture bottles;

[0044] (2) Add distilled water to the sugar-free and agar-free MS medium to make a 2 g / L solution, boil it, add 0.2 g / L sucrose, 0.01 g / L chlorobromoisocyanuric acid, 0.2 g / L potassium rosinate, 0.1 g / L potassium acetate, 0.2 g / L salicylic acid, 0.1 g / L glutaraldehyde, and 1.5 mg / L indolebutyric acid, and stir to prepare liquid medium A. Liquid medium A needs to be prepared and used immediately;

[0045] (3) adding the liquid culture medium A in step (2) to the tissue culture bottle to form a liquid layer with a thickness of 3 mm, adding the disinfected Venus flytrap leaves in step (1), sealing, and culturing for 12 days under a light intensity of 1000 lux and a wavelength of 650 nm red light to obtain Venus flytrap leaves with dense adventitious buds;

[0046] (4) Add distilled water to the sugar-free and agar-free MS medium to make a 2 g / L solution, boil it, add 0.2 g / L sucrose, 0.01 g / L chlorobromoisocyanuric acid, 0.2 g / L potassium rosinate, 0.1 g / L potassium acetate, 0.2 g / L salicylic acid, 0.1 g / L glutaraldehyde, and 1 mg / L indolebutyric acid, and stir to prepare liquid medium B. Liquid medium B needs to be prepared and used immediately;

[0047] (5) replacing the culture medium in the tissue culture bottle with the liquid culture medium B in step (4) to form a liquid layer with a thickness of 3 mm, adding the Venus flytrap leaves with dense adventitious buds in step (3), sealing, and culturing for 14 days under a light intensity of 1000 lux, 650 nm red light and 2000 lux, 460 nm blue light, to obtain Venus flytrap leaves with roots;

[0048] (6) Take out the leaf of the Venus flytrap with roots from step (5), clean it and plant it in a sphagnum moss substrate, and cultivate it under natural light with a light intensity of 2000 lux. Control the humidity of the environment to be 80%. Cultivate it for 5 days. When the bud in the center of the plant grows, the open tissue culture of the Venus flytrap is completed. The Venus flytrap seedlings are as follows: Figure 2 shown.

[0049] Comparative Example 1

[0050] The invention discloses a water-insertion propagation method for Venus flytrap.

[0051] The following steps are involved:

[0052] (1) Select healthy Venus flytrap leaves with buds, wash them with clean water and set aside;

[0053] (2) Tear up the peat moss and spread it flat in the container;

[0054] (3) Seal the container and place it in a place with diffuse light for 30 to 45 days. When the petiole buds are divided and roots are produced, take them out and place them in a greenhouse with 70% humidity for 10 days to train the seedlings.

[0055] Therefore, the actual cultivation time of the present invention is 31 days, while the comparative example 1 actually requires 55 days for cultivation, which indicates that the open tissue culture method of Venus flytrap provided in the embodiment 1 of the present invention has a shorter tissue culture time and a better breeding effect.

[0056] The preferred specific embodiments of the present invention are described in detail above. It should be understood that a person skilled in the art can make many modifications and changes based on the concept of the present invention without creative work. Therefore, any technical solution obtained by any modification, equivalent replacement, improvement, etc. made by a person skilled in the art based on the concept of the present invention through logical analysis, reasoning or limited experiments on the basis of the prior art shall be within the scope of protection determined by the claims.

Claims

1. A method for the open tissue culture of Venus flytrap, characterized in that: The following steps are involved: (1) selecting a leaf of Venus flytrap, washing it, and then soaking and disinfecting it in 0.1-0.2% chlorobromoisocyanuric acid to obtain a disinfected leaf of Venus flytrap; (2) Add water to a 1.5-2 g / L solution of sugar-free and agar-free MS medium, boil it, add 0.1-3 g / L sucrose, 0.01-1 g / L chlorobromoisocyanuric acid, 0.2-0.5 g / L potassium rosinate, 0.05-0.1 g / L potassium acetate, 0.1-0.2 g / L salicylic acid, 0.1-0.2 g / L glutaraldehyde, and 1.5-3 mg / L cytokinin, and stir to prepare a liquid medium A; (3) adding the liquid culture medium A described in step (2) into a container to form a liquid layer with a thickness of 2 to 3 mm, adding the disinfected Venus flytrap leaves described in step (1), sealing, and culturing under 950 to 1050 lux red light for 10 to 15 days to obtain Venus flytrap leaves with dense adventitious buds; (4) Add water to the sugar-free and agar-free MS medium to make a 1.5-2 g / L solution, boil it, add 0.2-0.3 g / L sucrose, 0.01-1 g / L chlorobromoisocyanuric acid, 0.2-0.5 g / L potassium rosinate, 0.05-0.1 g / L potassium acetate, 0.1-0.2 g / L salicylic acid, 0.1-0.2 g / L glutaraldehyde, and 0.5-1 mg / L indolebutyric acid, and stir to prepare a liquid medium B; (5) replacing the culture medium in the container with the liquid culture medium B described in step (4) to form a liquid layer with a thickness of 2 to 3 mm, adding the Venus flytrap leaves with dense adventitious buds described in step (3), sealing, and culturing for 10 to 15 days under the co-illumination of 950 to 1050 lux red light and 1950 to 2050 lux blue light to obtain Venus flytrap leaves with roots; (6) Take out the leaf of the Venus flytrap with roots described in step (5), clean it, and plant it in a substrate, and cultivate it under natural light for 4 to 6 days to complete the open tissue culture of the Venus flytrap.

2. The tissue culture method according to claim 1, characterized in that In step (1), the Venus flytrap leaf is a healthy Venus flytrap leaf with buds.

3. The tissue culture method according to claim 1, characterized in that In step (1), the soaking and disinfection time is 20 to 40 minutes.

4. The tissue culture method according to claim 1, characterized in that In step (2), the cytokinin includes at least one of indolebutyric acid.

5. The tissue culture method according to claim 1, characterized in that: In step (3), the wavelength of the red light is 620-720 nm.

6. The tissue culture method according to claim 1, characterized in that: In step (5), the wavelength of the blue light is 400 to 520 nm.

7. The tissue culture method according to claim 1, characterized in that: In step (6), the substrate is sphagnum moss.

8. The tissue culture method according to claim 1, characterized in that: In step (6), the illumination intensity of the natural light is 2000-2500 lux.

9. The tissue culture method according to claim 1, characterized in that: In step (6), the humidity of the seedling training culture is 70-90%.

10. Use of the tissue culture method according to any one of claims 1 to 9 in the rapid propagation of Venus flytraps.

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