Culture medium capable of prolonging viewing period of micro landscape of tissue culture bottle

Abstract

The invention relates to a culture medium capable of prolonging the ornamental period of a tissue culture bottle microlandscape, the culture medium is prepared by adding a plant growth regulator, sucrose and agar into an MS culture medium, and the pH value of the culture medium is 5.6-6.0. The requirements of plant growth on moisture and nutrition can be met, and meanwhile, a proper amount of hormone for plant growth and the like are added into the culture medium, so that the ornamental period can be prolonged. The culture medium can be used for tissue culture of micro-landscape plants, and compared with an existing method for manufacturing micro-landscapes through fresh cut flowers, matched leaves and the like, the ornamental period of the micro-landscapes can be remarkably prolonged, and meanwhile the cost is reduced. Test results show that the ornamental period of the obtained product can reach 8 months or even longer.

Description

Technical Field

[0001] The invention relates to the field of plant culture medium, in particular to a culture medium capable of prolonging the viewing period of micro-landscape in tissue culture bottles. Background Art

[0002] Micro landscape is to combine various elements such as plants, fences, gravel, cartoon characters and animals in a bottle to form a specific scene. Traditional micro landscape adopts soil cultivation, and its normal growth depends on regular watering, fertilization, soil replacement and pest control. However, the management and maintenance of this traditional potted plant is cumbersome, time-consuming and laborious.

[0003] Tissue culture bottle micro landscape is a technology that cultivates plants in tissue culture bottles to form a micro landscape. It combines plant tissue culture and micro landscape design concepts, and has the characteristics of high ornamental value and small space occupation. This kind of micro landscape has many advantages. On the one hand, it solves the problem of time-consuming and labor-intensive management and maintenance of traditional potted plants; on the other hand, tissue culture bottles of different shapes and sizes can be used, which improves the ornamental value.

[0004] Micro-landscapes in tissue culture bottles usually contain a variety of plants, and different plants have significant differences in their demand for and absorption capacity for nutrient elements. Universal culture media cannot adapt well to this diversity, resulting in poor growth of some plants due to excess or insufficient nutrition, affecting the ornamental cycle and ornamental value. At the same time, different plants have different sensitivities to growth regulators. In a complex system such as micro-landscapes in tissue culture bottles that contains a variety of plants, it is difficult to accurately control the concentration and duration of action of growth regulators to meet the optimal growth needs of each plant. If the concentration is too high, it may cause plant growth deformities or other abnormal phenomena; if the concentration is too low, the expected growth promotion effect cannot be achieved.

[0005] The culture environment of the micro-landscape in tissue culture bottles is relatively closed and easily contaminated by microorganisms. The existing culture medium itself does not have sufficient antibacterial ability, and during the long-term culture process, microorganisms can easily invade from the outside or breed inside the culture medium. Once microbial contamination, such as bacterial or fungal contamination, occurs, plant tissues may rot and necrotize, causing the entire micro-landscape to lose its ornamental value.

[0006] Moreover, most of the existing tissue culture bottle micro-landscape technologies are to sterilize cut flowers and leaves (with higher costs) purchased from the market and then put them into tissue culture bottles, rather than using plant tissue culture. This technology is easily contaminated and has not established a stable regeneration system. Tissue culture containers have problems with insufficient space and nutrition. These factors directly restrict the survival time of plants, which in turn greatly limits the ornamental value and viewing time of tissue culture bottle micro-landscapes, making it difficult to achieve ideal ornamental effects and duration. Summary of the invention

[0007] In view of the above-mentioned deficiencies in the prior art, the present invention proposes a culture medium that can prolong the viewing period of micro-landscapes in tissue culture bottles. The culture medium can construct a stable regeneration system by culturing the tissues of micro-landscape plants in tissue culture bottles, thereby prolonging the viewing period of micro-landscapes in tissue culture bottles.

[0008] The specific technical scheme of the present invention is as follows: a culture medium capable of prolonging the viewing period of micro-landscape in tissue culture bottles, wherein the culture medium is prepared by adding plant growth regulators, sucrose and agar to MS culture medium, and the pH of the culture medium is 5.6-6.0.

[0009] MS culture medium can provide basic nutrition for plants, sucrose can control plant flowering, and plant growth agents can regulate plant rooting and flowering. Under pH conditions suitable for plant growth, these ingredients work together to achieve continuous growth and flowering effect regulation of micro-landscape plants in tissue culture bottles, build a stable regeneration system, extend the viewing period of tissue culture micro-landscapes, and enhance their ornamental value.

[0010] Preferably, the plant growth regulator is at least one of 6-BA and NAA. 6-BA is a synthetic cytokinin that can promote plant proliferation and simulate the vibrant scene of spring and summer. The plant hormone NAA can promote rooting and inhibit the proliferation of micro-landscape plants in the tissue culture bottle. The two cooperate with each other to maintain the state of the micro-landscape in the tissue culture bottle for a long time.

[0011] Further preferably, the culture medium is prepared by adding 0.1-1 mg / L 6-BA, 0.1-0.2 mg / L NAA, 20-40 g / L sucrose and 6.5-7.5 g / L agar to 1 / 4-1 / 2 MS culture medium. The obtained culture medium is used for micro-landscape tissue culture, which can make the product's ornamental period up to 8 months or even longer.

[0012] Preferably, the concentration ratio of 6-BA to NAA is 2 to 3:1. Within this ratio range, the state of the micro-landscape plants can be preserved for a longer time.

[0013] Preferably, the culture medium also contains pigments to increase the ornamental value.

[0014] In another aspect of the present invention, there is also provided a method for extending the viewing period of micro-landscape in a tissue culture bottle, comprising the following steps:

[0015] 1) Prepare the culture medium: place the culture medium that can prolong the viewing period of the micro-landscape in the tissue culture bottle into the tissue culture bottle and sterilize it;

[0016] 2) Inoculation of tissue culture materials: In a sterile environment, inoculate the micro-landscape plant tissue culture materials on the culture medium according to the set method;

[0017] 3) Tissue culture: seal the tissue culture bottle, and then culture at a temperature of 24±2°C, a light intensity of 2000-3000Lx, and a light duration of 16 hours / day.

[0018] This method can easily build a stable system and prolong the viewing period of the micro-landscape in the tissue culture bottle by constructing a sterile initial environment, matching with a suitable culture medium, and selecting tissue materials to culture the micro-landscape in a closed manner. It can also greatly reduce the dependence on fresh cut flowers and leaves in the market, and greatly reduce the cost of micro-landscape production.

[0019] Preferably, the tissue culture bottle is a borosilicate glass bottle. The reason is that it is resistant to high temperatures and is suitable for high-temperature sterilization operations. In addition, the mouth of this bottle is moderately sized, easy to seal, convenient for moving the prepared plant tissue into the bottle, and can ensure a low contamination rate. Furthermore, it is easy to shape, and borosilicate glass bottles of various shapes and sizes can meet the needs of various plants to fully display the states of different growth periods.

[0020] Preferably, the sterilization treatment temperature is 121°C.

[0021] Select micro-plants for tissue culture. Micro-plants are generally small in size, beautiful in shape, highly ornamental in color and relatively long in life cycle. Flowers or leaves. The shape is exquisite and small, deeply loved by people, and has broad application prospects in interior decoration, small landscape construction, etc. Moreover, micro-plants can complete the entire life cycle in the tissue culture bottle, and can even bloom in the tissue culture bottle, thereby achieving the purpose of extending the viewing period. In addition, plants with consistent growth temperature can be selected to build micro-landscapes: avoid the impact of temperature on micro-landscapes and reduce the regulation of plant growth temperature by micro-landscape holders, reduce the maintenance cost of micro-landscape holders, and facilitate consumers in different regions and seasons. Preferably, various micro-plants with established regeneration systems in the laboratory can be selected to design landscapes, including at least one of micro-roses, lithops, succulents, sundews, begonias, Venus flytraps, pansy, and pitcher plants. These plants are deeply loved by the public, small and exquisite, and have a long life cycle, which can prevent the occurrence of browning and withering of some micro-landscapes in the tissue culture bottle. Before the nutrients in the culture medium are exhausted, these plants can be inoculated into a new culture medium together, which will not affect the subsequent ornamental effect.

[0022] Preferably, a light-transmitting plastic bottle cap is used to seal instead of the traditional sealing film. The sealing film used for the micro-landscape of the tissue culture bottle in the prior art is light-transmitting and breathable, but in the actual operation process, the sealing film is easily damaged and leaks due to friction during transportation, which leads to contamination in the tissue culture bottle and damages the micro-landscape. During plant tissue culture, the entire culture system needs to remain aseptic, and the micro-landscape in the tissue culture bottle constructed by the present application is also aseptic at the beginning, but the subsequent environment is more complicated. To ensure that the micro-landscape in the tissue culture bottle is in an aseptic state for a long time, even if it is in various complex environments, it is not affected, and the sealing film selects an airtight plastic bottle cap to prevent external gas from entering the tissue culture bottle, and prevents the aseptic state of the micro-landscape in the tissue culture bottle from being changed. At the same time, in order not to affect the normal growth of the micro-landscape plant in the tissue culture bottle, the sealing film selects a light-transmitting plastic bottle cap to prevent the light conditions received by the micro-landscape plant in the tissue culture bottle from being changed due to the use of a light-transmitting sealing film, and then avoids adverse effects on the growth of the micro-landscape plant in the tissue culture bottle.

[0023] The beneficial effects of the present invention are as follows: the present invention provides a culture medium formula that can improve the viewing period of micro-landscape in tissue culture bottles, which can not only meet the needs of plant growth for water and nutrition, but also add an appropriate amount of plant growth hormones to the culture medium, which can extend the viewing period. In combination with the culture medium, a method for extending the viewing period of micro-landscape in tissue culture bottles is provided, which can significantly extend the viewing period of micro-landscape compared to the existing methods of making micro-landscapes by fresh cut flowers, matching leaves, etc., while reducing costs. The test results show that the viewing period of the obtained product can reach 8 months or even longer, which can make up for the shortcomings of the existing products on the market, such as the single color, time-consuming and labor-intensive management and maintenance of traditional potted plants, and the short survival time, and fill the market gap. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a flow chart of the method for prolonging the viewing period of micro-landscape in tissue culture bottles of the present invention;

[0025] Figure 2 This is a schematic diagram of the micro landscape of the tissue culture bottle of Example 1 of the present invention;

[0026] Figure 3 This is a schematic diagram of the micro landscape of the tissue culture bottle of Example 2 of the present invention;

[0027] Figure 4 This is a schematic diagram of the micro landscape of the tissue culture bottle of Example 3 of the present invention;

[0028] Figure 5 This is a schematic diagram of the micro landscape of the tissue culture bottle of Example 4 of the present invention;

[0029] Figure 6 This is a schematic diagram of the micro landscape of the tissue culture bottle of Comparative Example 1 of the present invention;

[0030] Figure 7 Schematic diagram of tissue culture bottles with different structures. DETAILED DESCRIPTION

[0031] The present invention is further described below with reference to specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0032] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0033] Example 1

[0034] A culture medium capable of extending the viewing period of a micro-landscape in a tissue culture bottle is prepared by adding 0.1 mg / L 6-BA, 0.2 mg / L NAA, 25 g / L sucrose and 6.5 g / L agar to a 1 / 2MS culture medium and adjusting the pH value to 5.6.

[0035] refer to Figure 1 , a method for extending the viewing period of micro-landscape in tissue culture bottles, comprising the following steps:

[0036] 1) Prepare the culture medium: Place the prepared culture medium in a borosilicate glass tissue culture bottle, then place it in an autoclave and sterilize it at 121°C for 20 minutes. After sterilization, take it out and wait for it to cool to room temperature.

[0037] 2) Inoculation of tissue culture materials: Select various micro-plants with established regeneration systems in the laboratory to design the landscape, including sterile seedlings of pitaya, miniature roses, and one or more sterile seedlings of lithops, succulents, sundews, begonias, Venus flytraps, butterfly traps, and pitcher plants. Put the cooled and solidified culture medium and the selected plant tissue culture materials into the clean bench and sterilize them with ultraviolet light for 20-30 minutes. After that, open the clean bench for air blowing for 5-10 minutes, and inoculate the tissue culture materials on the culture medium according to the set method.

[0038] 3) Tissue culture: Seal the tissue culture bottle with a transparent plastic bottle cap, and then culture at a temperature of 24±2°C, a light intensity of 2000-3000Lx, and a light duration of 16 hours / day.

[0039] The micro landscape of the tissue culture bottle obtained in Example 1 is as follows Figure 2 shown.

[0040] Example 2

[0041] A culture medium capable of extending the viewing period of a micro-landscape in a tissue culture bottle is prepared by adding 0.2 mg / L 6-BA, 0.1 mg / L NAA, 25 g / L sucrose and 6.5 g / L agar to a 1 / 2MS culture medium and adjusting the pH value to 5.6.

[0042] refer to Figure 1 , a method for extending the viewing period of micro-landscape in tissue culture bottles, comprising the following steps:

[0043] 1) Prepare the culture medium: Place the prepared culture medium in a borosilicate glass tissue culture bottle, then place it in an autoclave and sterilize it at 121°C for 20 minutes. After sterilization, take it out and wait for it to cool to room temperature.

[0044] 2) Inoculation of tissue culture materials: Select various micro-plants with established regeneration systems in the laboratory to design the landscape, including sterile seedlings of pitaya, miniature roses, and one or more sterile seedlings of lithops, succulents, sundews, begonias, Venus flytraps, butterfly traps, and pitcher plants. Put the cooled and solidified culture medium and the selected plant tissue culture materials into the clean bench and sterilize them with ultraviolet light for 20-30 minutes. After that, open the clean bench for air blowing for 5-10 minutes, and inoculate the tissue culture materials on the culture medium according to the set method.

[0045] 3) Tissue culture: Seal the tissue culture bottle with a transparent plastic bottle cap, and then culture at a temperature of 24±2°C, a light intensity of 2000-3000Lx, and a light duration of 16 hours / day.

[0046] The micro landscape of the tissue culture bottle obtained in Example 2 is as follows Figure 3 shown.

[0047] Example 3

[0048] A culture medium capable of extending the viewing period of a micro-landscape in a tissue culture bottle is prepared by adding 0.4 mg / L 6-BA, 0.2 mg / L NAA, 30 g / L sucrose and 7.5 g / L agar to a 1 / 4MS culture medium and adjusting the pH value to 5.8.

[0049] refer to Figure 1 , a method for extending the viewing period of micro-landscape in tissue culture bottles, comprising the following steps:

[0050] 1) Prepare the culture medium: Place the prepared culture medium in a borosilicate glass tissue culture bottle, then place it in an autoclave and sterilize it at 121°C for 20 minutes. After sterilization, take it out and wait for it to cool to room temperature.

[0051] 2) Inoculation of tissue culture materials: Select various micro-plants with established regeneration systems in the laboratory to design the landscape, including sterile seedlings of pitaya, miniature roses, and one or more sterile seedlings of lithops, succulents, sundews, begonias, Venus flytraps, butterfly traps, and pitcher plants. Put the cooled and solidified culture medium and the selected plant tissue culture materials into the clean bench and sterilize them with ultraviolet light for 20-30 minutes. After that, open the clean bench for air blowing for 5-10 minutes, and inoculate the tissue culture materials on the culture medium according to the set method.

[0052] 3) Tissue culture: Seal the tissue culture bottle with a transparent plastic bottle cap, and then culture at a temperature of 24±2°C, a light intensity of 2000-3000Lx, and a light duration of 16 hours / day.

[0053] The micro landscape of the tissue culture bottle obtained in Example 3 is as follows Figure 4 shown.

[0054] Example 4

[0055] A culture medium capable of extending the viewing period of a micro-landscape in a tissue culture bottle is prepared by adding 0.6 mg / L 6-BA, 0.2 mg / L NAA, 30 g / L sucrose and 7.5 g / L agar to a 1 / 4MS culture medium and adjusting the pH value to 6.0.

[0056] refer to Figure 1 , a method for extending the viewing period of micro-landscape in tissue culture bottles, comprising the following steps:

[0057] 1) Prepare the culture medium: Place the prepared culture medium in a borosilicate glass tissue culture bottle, then place it in an autoclave and sterilize it at 121°C for 20 minutes. After sterilization, take it out and wait for it to cool to room temperature.

[0058] 2) Inoculation of tissue culture materials: Select various micro-plants with established regeneration systems in the laboratory to design the landscape, including sterile seedlings of pitaya, miniature roses, and one or more sterile seedlings of lithops, succulents, sundews, begonias, Venus flytraps, butterfly traps, and pitcher plants. Put the cooled and solidified culture medium and the selected plant tissue culture materials into the clean bench and sterilize them with ultraviolet light for 20-30 minutes. After that, open the clean bench for airing for 5-10 minutes, and inoculate the tissue culture materials on the culture medium according to the set method; finally, use a plastic bottle cap to seal the inoculated tissue culture bottle.

[0059] 3) Tissue culture: Seal the tissue culture bottle, and then culture it at a temperature of 24±2℃, a light intensity of 2000-3000Lx, and a light duration of 16 hours / day. It will bloom in about 10 days after inoculation. Figure 5shown.

[0060] Comparative Example 1

[0061] This comparative example is a method for constructing an artificial combined landscape in the related art. In this method, the culture medium of the combined potted plants is sphagnum moss, and the container is an acrylic landscaping tank.

[0062] Its construction specifically includes the following steps: First, the plants are prepared. The selected plants are all plant seedlings grown in natural environments, such as Phalaenopsis, butterfly orchids, sphagnum moss and succulents. Inoculate to construct the combined potted plants, fill the sphagnum moss in a slope shape to 1 / 3-1 / 2 of the height of the acrylic landscaping tank, and then arrange and combine according to the different colors of the plants and the natural plant growth style, and finally move the prepared succulents, butterfly orchids and other plants into the flowerpots to construct the combined potted plants. The maintenance conditions of the combined potted plants are a temperature of 24±2℃, a humidity of 85%, a light intensity of 2000-3000Lx, and a light duration of 16h / d.

[0063] like Figure 6 As shown, if there is no water and fertilizer management for a period of time, the plants in the micro landscape will wither and even die.

[0064] Comparative Example 2

[0065] This comparative example provides a culture medium for micro-landscape, which is prepared by adding 0 mg / L 6-BA, 0.2 mg / L NAA, 25 g / L sucrose and 7.5 g / L agar to MS culture medium and adjusting the pH value to 5.6.

[0066] The obtained culture medium is used to construct a micro-landscape in a tissue culture bottle, which specifically includes the following steps:

[0067] 1) Prepare the culture medium: Place the prepared culture medium in a borosilicate glass tissue culture bottle, then place it in an autoclave and sterilize it at 121°C for 20 minutes. After sterilization, take it out and wait for it to cool to room temperature.

[0068] 2) Plant inoculation: linear plants, loose plants and clumps of plants purchased from the market. Put the cooled and solidified culture medium and the selected plants into the clean bench and sterilize them with ultraviolet light for 20-30 minutes. After that, open the clean bench to blow air for 5-10 minutes, and inoculate the plants on the culture medium according to the set method.

[0069] 3) Cultivation: Seal the inoculated tissue culture bottle with a sealing film, and then cultivate at a temperature of 24±2°C, a light intensity of 1500-3000Lx, and a light duration of 16 hours / day.

[0070] Table 1 Parameters of Examples and Comparative Examples

[0071]

[0072] As shown in Table 1, through the parameters of the maintenance process of the micro-landscape constructed by the embodiments and comparative examples, it can be known that the micro-landscape in the tissue culture bottle constructed by the method of the present application does not need watering or fertilization during the maintenance period; at the same time, because the micro-plants with exquisite and delicate characteristics that have established a regeneration system are used, the ornamental value can be greatly improved compared with comparative example 1. The micro-plants used are exquisite and delicate micro-plants that are now popular with the public, such as the popular plants such as Venus flytraps, Venus flytraps, and sundews, which have higher ornamental value than comparative example 1. During the maintenance process, the micro-landscape in the landscaping tank of comparative example 1 needs to be watered once a week and fertilized once a month. It is more susceptible to the invasion of pests and diseases. In Examples 1-4 of the present invention, micro-plant tissue culture is used. Because of the closed maintenance, there is no need to worry about the influence of pests and diseases, and it also has a longer viewing period.

[0073] Moreover, it can be seen from the comparison that adding 6-BA and NAA at the same time is more conducive to prolonging the viewing period. Moreover, the viewing period can be further delayed by adjusting the dosage ratio of 6-BA and NAA in the culture medium. Generally speaking, the viewing period increases with the increase of dosage, and when the concentration ratio of 6-BA to NAA is 2-3:1, it has a better viewing period.

[0074] The high borosilicate glass tissue culture bottles used in the embodiments of the present invention can be selected from high borosilicate glass bottles of different shapes and sizes. Figure 2-5 The structure shown in Figure 7 Various structural types in. High borosilicate glass bottles of various shapes and sizes can meet the needs of various plants to fully display the states of different growth periods.

[0075] The technical features of the above-mentioned embodiments can be combined arbitrarily. To make the description concise, all possible combinations of the technical features in the above-mentioned embodiments are not described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification. 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, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the attached claims.

Claims

1. A culture medium capable of extending the viewing period of micro-landscape in tissue culture bottles, characterized in that: The culture medium is prepared by adding plant growth regulator, sucrose and agar into MS culture medium, and the pH value of the culture medium is 5.6-6.

0.

2. The culture medium according to claim 1, characterized in that The plant growth regulator is at least one of 6-BA and NAA.

3. The culture medium according to claim 2, characterized in that The culture medium is prepared by adding 0.1-1 mg / L 6-BA, 0.1-0.3 mg / L NAA, 20-40 g / L sucrose and 6.5-7.5 g / L agar to 1 / 4-1 / 2 MS culture medium.

4. The culture medium according to claim 3, characterized in that The concentration ratio of 6-BA to NAA is 2 to 3:

1.

5. The culture medium according to any one of claims 1 to 4, characterized in that The culture medium also contains pigment.

6. A method for extending the viewing period of micro-landscape in tissue culture bottles, characterized in that: The steps include: 1) Preparing a culture medium: placing the culture medium according to any one of claims 1 to 5 into a tissue culture bottle and sterilizing it; 2) Inoculating tissue culture materials: In a sterile environment, inoculate micro-plant tissue culture materials on the culture medium according to the set method; 3) Tissue culture: seal the tissue culture bottle, and then culture at a temperature of 24±2°C, a light intensity of 2000-3000Lx, and a light duration of 16 hours / day.

7. The method according to claim 6, characterized in that The tissue culture bottle is a high borosilicate glass bottle.

8. The method according to claim 6, characterized in that The sterilization temperature is 121°C.

9. The method according to claim 6, characterized in that The micro-plants are micro-plants for which a regeneration system has been established in the laboratory, including at least one of micro-roses, lithops, succulents, sundews, begonias, Venus flytraps, pansy, and pitcher plants.

10. The method according to claim 6, characterized in that Use a light-transmitting plastic bottle cap for sealing.

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