Warped plate type liquid culture device and culture method

Abstract

The invention discloses a seesaw type liquid culture device. The device comprises a seesaw type shaking table; the culture box is arranged on the seesaw type shaking table, a plurality of partition plates are arranged in the culture box, and the culture box is divided into a plurality of culture spaces by the partition plates; the culture bottle is correspondingly arranged in the culture space of the culture box, the culture bottle comprises an outer bottle and an inner bottle, the outer bottle is used for containing a liquid culture solution, the inner bottle is arranged in the outer bottle, a supporting part is arranged at the bottom of the inner bottle, a spacing distance exists between the inner bottle and the bottom of the outer bottle, a plurality of water permeable holes are formed in the bottom of the inner bottle, and the water permeable holes are communicated with the outer bottle. The inner surface is provided with at least one partition plate and forms a culture area. According to the seesaw type liquid culture device, the culture box is arranged on the seesaw type shaking table, the shaking table shakes regularly, and culture materials are blocked by the partition plates, so that the culture materials cannot be continuously soaked in a culture solution, but can be periodically supplied with nutrients, and tissue culture materials can grow quickly.

Description

Technical Field

[0001] The invention relates to the technical field of plant tissue culture, and in particular to a seesaw type liquid culture device and a culture method. Background Art

[0002] In the process of plant tissue culture industrialization, the common culture methods are solid culture and liquid culture. Solid culture is currently more widely used, while liquid culture is not widely used because it can easily lead to restricted respiration of plant materials and hindered growth. However, liquid culture has a fast reproduction speed, can make the materials obtain better nutrition, and can more accurately control the temperature of the solution and the concentration of nutrients. In view of the problem that liquid culture can easily lead to restricted respiration of plant materials, various solutions have been proposed in production, among which intermittent immersion culture is a liquid culture method that has been reported more frequently. This method can periodically immerse tissue culture materials in culture solution and requires an air pump device, a timing device, a conduction device, and a bacterial filter membrane. The devices are numerous and complex, and the key is to have a good sealing effect. Once there is a leak, it cannot be achieved. This shortcoming makes it difficult to apply and promote intermittent immersion culture in production.

[0003] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0004] The object of the present invention is to provide a seesaw type liquid culture device, so as to overcome the shortcomings of the liquid culture device such as complex structure and easy leakage.

[0005] Another object of the present invention is to provide a method for culturing using a seesaw type liquid culture device.

[0006] To achieve the above-mentioned purpose, the present invention provides a seesaw liquid culture device, comprising: a seesaw shaker; an incubator, which is arranged on the seesaw shaker, and a plurality of partitions are arranged in the incubator, and the plurality of partitions divide the incubator into a plurality of culture spaces; and a culture bottle, which is placed in the culture space of the incubator accordingly, and the culture bottle comprises an outer bottle and an inner bottle, the outer bottle is used to hold liquid culture solution, the inner bottle is placed in the outer bottle, a support portion is provided at the bottom of the inner bottle, and there is a spacing distance between the inner bottle and the bottom of the outer bottle, a plurality of water permeable holes are provided at the bottom of the inner bottle, and at least one partition is provided on the inner surface to form a culture area.

[0007] Preferably, in the above technical solution, the incubator is a sealed structure, provided with an air inlet and air holes, and a breathable filter membrane is provided at the air holes.

[0008] Preferably, in the above technical solution, the partitions in the incubator are arranged crosswise with each other.

[0009] Preferably, in the above technical solution, a plurality of water-permeable holes are provided on the surrounding bottle walls of the inner bottle close to the bottle bottom.

[0010] Preferably, in the above technical solution, the bottle walls around the inner bottle are fence-shaped.

[0011] A seesaw liquid culture method, using the above-mentioned culture device for culture, comprises the following steps:

[0012] (1) Preparation of nutrient solution: Dispense the prepared nutrient solution into the inner bottle of the culture bottle, add a quantity of culture solution just enough to cover the bottom of the inner bottle, then place in an incubator, seal, and sterilize;

[0013] (2) Inoculation: Select seedlings and place them into the inner bottle of a culture bottle, then place the culture bottle into the culture space of an incubator and cover the incubator with a lid;

[0014] (3) Cultivation: Place the incubator on a seesaw shaker, connect the air inlet of the incubator to the air inlet pipe, introduce CO2, start the shaker, and start culturing.

[0015] Preferably, in the above technical solution, the culture conditions of step (3) are: culture temperature 23-27°C, light intensity 2400-3200lx, light cycle 10-12h, and culture for 50-70 days.

[0016] Preferably, in the above technical solution, the rocking frequency of the seesaw rocking table is 40-70 times / min.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] (1) The seesaw liquid culture device of the present invention is used for liquid culture. Compared with solid culture, liquid culture can more accurately control the temperature of the solution and the concentration of nutrients, and the propagation speed is faster. The device places the incubator on a seesaw shaker, which shakes regularly. The culture material is blocked by a partition and will not be continuously immersed in the culture solution, but it can obtain nutrition periodically, so that the tissue culture material can grow rapidly. The use of periodic immersion culture solves the problem of limited respiration in liquid culture, and provides nutrition while ensuring the normal respiration of the culture material.

[0019] (2) The liquid culture method of the present invention uses CO2 as a carbon source and introduces CO2 into the culture box. Sugar needs to be added to the culture medium of conventional solid culture as a carbon source for the culture material. Therefore, the tissue culture seedlings of solid culture generally have weak respiration capacity, and the sugar content in the culture medium is also prone to pollution. The present invention uses CO2 as a carbon source, allowing the material to photosynthesize autonomously, and has a strong respiration capacity, and a high survival rate of transplantation in the later stage. After sugar is not added to the nutrient solution, the pollution rate is also reduced. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 is a schematic structural diagram of a culture box in a seesaw type liquid culture device according to the present invention;

[0021] Figure 2 It is a schematic diagram of the structure of a culture bottle in a seesaw type liquid culture device according to the present invention. DETAILED DESCRIPTION

[0022] The specific implementation modes of the present invention are described in detail below in conjunction with the accompanying drawings, but it should be understood that the protection scope of the present invention is not limited by the specific implementation modes.

[0023] Unless explicitly stated otherwise, throughout the specification and claims, the term “comprise” or variations such as “include” or “comprising”, etc., will be understood to include the stated elements or components but not to exclude other elements or components.

[0024] like Figure 1 to Figure 2 As shown, a seesaw liquid culture device according to a specific embodiment of the present invention comprises a seesaw shaker, an incubator 1 and a culture bottle 2. When in use, the seedlings to be cultured and the liquid culture solution are placed in the culture bottle 2, the culture bottle 2 is placed in the incubator 1, and the incubator 1 is placed on the seesaw shaker, and the shaker swings regularly for culture.

[0025] The specific structure is as follows: the incubator 1 is placed on a seesaw shaker (not shown) when in use. A plurality of partitions 11 are provided in the incubator 1. In this embodiment, two partitions are provided. The two partitions 11 are arranged perpendicularly and crosswise to divide the incubator into four culture spaces. The incubator 1 is a sealed structure, provided with an air inlet 12 and an air vent 13, and a breathable filter membrane is provided at the air vent. The air inlet 12 is connected to the air inlet pipe 14, and CO2 gas is introduced into the incubator 1. The culture bottle 2 is placed in the culture space of the incubator. The culture bottle 2 includes an inner bottle 21 and an outer bottle 22. The outer bottle 22 is an open structure for holding liquid culture fluid. The inner bottle and the outer bottle of the culture bottle 2 are both made of high temperature resistant plastic material. The outer bottle 22 is cylindrical and is sized to fit into the small cube culture space area of ​​the incubator. A support portion 23 is provided at the bottom of the inner bottle 21. The support portion 23 is a support rod. Four support rods are provided at the bottom of the inner bottle 21. The inner bottle 21 is placed in the outer bottle 22. The diameter of the inner bottle 21 is 0.5 cm smaller than that of the outer bottle 22. Since a support rod is provided at the bottom of the inner bottle, there is a spacing of 1 cm between the bottom of the inner bottle 21 and the bottom of the outer bottle 22. The bottom of the inner bottle 21 is provided with a plurality of water-permeable holes so that liquid culture medium can enter the inner bottle. In this embodiment, a water-permeable filter screen is laid at the bottom of the inner bottle 21. Preferably, a plurality of water-permeable holes are provided on the surrounding walls of the inner bottle 21 near the bottom. The surrounding walls of the inner bottle are fence-shaped. It is convenient for the liquid culture medium to enter the inner bottle. At least one partition 24 is provided on the inner surface of the bottom of the inner bottle 21, and one partition is provided in this embodiment. The partition 24 separates the inner bottle into two culture areas.

[0026] The method of using the seesaw liquid culture device of the present invention is as follows: during culture, first pour the prepared nutrient solution into the inner bottle 21, the volume is preferably just enough to soak the bottom of the inner bottle 21, then place it in the incubator 1, cover it with a lid, and sterilize it at high temperature. In the inoculation room, place the inoculated material in two areas of the inner bottle 21. After the inoculation is completed, directly put the culture bottle 2 into the incubator 1, rotate and adjust the culture bottle 2 so that the partition 24 is perpendicular to the shaker to ensure that the liquid can shake left and right, then cover the lid of the incubator 1 and connect the CO2 ventilation tube. Place the incubator on the seesaw shaker, start the shaker, and shake at a frequency of 50 times / min. The culture solution shakes left and right like waves, and the culture material will not be continuously immersed in the culture solution because it is blocked by the partition, but it can obtain nutrient supply periodically, so that the tissue culture material can grow rapidly.

[0027] Example 1

[0028] The seesaw liquid culture device is used to culture Dendrobium officinale, and the culture method comprises the following steps:

[0029] (1) Nutrient solution preparation: MS basic culture medium + NAA 1.5 mg / L + banana juice 30 g / L, pH 5.6, no sugar and no agar. Mix well and dispense into culture bottles, 60 ml per bottle, just enough to cover the bottom of the inner bottle. After dispensing, put into the incubator, 4 per box, cover the incubator lid, sterilize in an autoclave, and cool for use.

[0030] (2) Inoculation: Select a bottle of healthy, uncontaminated Dendrobium officinale seedlings, take out the seedlings in a clean bench, separate them into individual plants and place them in the inner bottle of a culture bottle, 20 plants on each side. After four bottles are connected, place them directly in the incubator, rotate and adjust the culture bottle so that the partition is perpendicular to the shaker, ensure that the liquid can shake left and right, and then cover the incubator lid.

[0031] (3) Cultivation: Bring the incubator to the culture room, connect the CO2 ventilation tube to the incubator input port, then place the incubator on a seesaw shaker, start the shaker, and shake at a frequency of 50 times / min. The temperature in the culture room is 23℃~27℃, the light intensity is 2400lx~3200lx, and the photoperiod is 10h~12h. After 60 days of cultivation, the seedlings become taller and stronger, and grow roots, and can be transplanted and planted.

[0032] Example 2

[0033] The ornamental fern is cultivated using the seesaw type liquid cultivation device, and the cultivation method comprises the following steps:

[0034] (1) Nutrient solution: 1 / 2MS basic culture solution + IBA 2.0mg / L + NAA 0.1mg / L + activated carbon 1.5g / L, pH 5.6, no sugar and no agar. Mix well and dispense into culture bottles, 60ml per bottle, just enough to cover the bottom of the inner bottle. After dispensing, put into the incubator, 4 per box, cover the incubator lid, sterilize in the autoclave, and cool for use.

[0035] (2) Inoculation: Select a bottle of healthy, uncontaminated ornamental fern seedlings. Take out the seedlings in a clean bench, separate them into individual plants, and place them in the inner bottle of a culture bottle, 20 plants on each side. After four bottles are connected, place them directly in the incubator. Turn and adjust the culture bottle so that the partition is perpendicular to the shaker, ensure that the liquid can shake left and right, and then cover the incubator lid.

[0036] (3) Cultivation: Bring the incubator to the culture room, connect the CO2 ventilation tube to the incubator input port, then place the incubator on a seesaw shaker, start the shaker, and shake at a frequency of 50 times / min. The temperature in the culture room is 23℃~27℃, the light intensity is 2400lx~3200lx, and the photoperiod is 10h~12h. After 60 days of cultivation, the seedlings become taller and stronger, and grow roots, and can be transplanted and planted.

[0037] Example 3

[0038] The jasmine is cultured using the seesaw type liquid culture device, and the culture method comprises the following steps:

[0039] (1) Nutrient solution preparation: 1 / 2MS basic culture solution + IBA 0.8mg / L, pH 5.6, no sugar and no agar. After mixing evenly, dispense into culture bottles, 60ml per bottle, just enough to immerse the bottom of the inner bottle. After dispensing, put into the incubator, 4 per box, cover the incubator lid, sterilize in the autoclave, and cool for use.

[0040] (2) Inoculation: Select a bottle of healthy, uncontaminated jasmine seedlings, take out the seedlings in a clean bench, cut off the lower half, leave 3 cm-5 cm of the upper half, and place them vertically on the edge of the inner bottle of the culture bottle, with 10 plants on each side. After connecting 4 bottles, place them directly in the incubator, rotate and adjust the culture bottle so that the partition is perpendicular to the shaker, ensure that the liquid can shake left and right, and then cover the incubator lid.

[0041] (3) Cultivation: Bring the incubator to the culture room, connect the CO2 ventilation tube to the incubator input port, then place the incubator on a seesaw shaker, start the shaker, and shake at a frequency of 50 times / min. The temperature in the culture room is 23℃~27℃, the light intensity is 2400lx~3200lx, and the photoperiod is 10h~12h. After 60 days of cultivation, the seedlings become taller and stronger, and grow roots, and can be transplanted and planted.

[0042] Control group

[0043] Dendrobium, ornamental fern and jasmine were cultured using the traditional solid culture method of the control group. The control group used traditional solid rooting culture, and the seedlings were inoculated into the corresponding culture medium containing agar and sugar. Among them, the culture medium of Dendrobium is: MS+NAA1.5mg / L+banana juice 30g / L+sugar 30g / L+agar 4.2g / L. The culture medium of ornamental fern is: 1 / 2MS+IBA2.0mg / L+NAA0.1mg / L+activated carbon 1.5g / L+sugar 30g / L+agar 4.2g / L. The culture medium of jasmine is: 1 / 2MS+IBA0.8mg / L+sugar 30g / L+agar 4.2g / L. The temperature of the culture room is 23℃~27℃, the light intensity is 2400lx~3200lx, the photoperiod is 10h~12h, and it is cultured for 60 days before transplanting.

[0044] The survival rates of seedlings transplanted after the rooting culture of the present invention and the traditional method are compared, as shown in Table 1.

[0045] Table 1

[0046]

[0047] As shown in Table 1, the transplant survival rate of dendrobium, ornamental fern and jasmine cultured by the seesaw liquid culture of the present invention is higher than that of the traditional solid rooting culture.

[0048] The foregoing description of specific exemplary embodiments of the present invention is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present invention to the precise form disclosed, and it is clear that many changes and variations can be made based on the above teachings. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present invention and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present invention and various different selections and changes. The scope of the present invention is intended to be limited by the claims and their equivalents.

Claims

1. A seesaw type liquid culture device, characterized in that: include: Rocker shaker; An incubator, which is arranged on the seesaw shaker, wherein a plurality of partitions are arranged in the incubator, and the plurality of partitions partition the incubator into a plurality of culture spaces; as well as A culture bottle is placed in the culture space of the culture box, the culture bottle includes an outer bottle and an inner bottle, the outer bottle is used to hold liquid culture solution, the inner bottle is placed in the outer bottle, a support portion is provided at the bottom of the inner bottle, and there is a distance between the inner bottle and the bottom of the outer bottle, a plurality of water permeable holes are provided at the bottom of the inner bottle, and at least one partition is provided on the inner surface to form a culture area.

2. The seesaw type liquid culture device according to claim 1, characterized in that: The incubator is a sealed structure, provided with an air inlet and air holes, and an air permeable filter membrane is provided at the air holes.

3. The seesaw type liquid culture device according to claim 1, characterized in that: The partitions in the incubator are arranged crosswise with each other.

4. The seesaw type liquid culture device according to claim 1, characterized in that: The inner bottle has a plurality of water-permeable holes arranged on the bottle wall around the bottle bottom.

5. The seesaw type liquid culture device according to claim 1, characterized in that: The bottle walls around the inner bottle are in a fence shape.

6. A seesaw liquid culture method, characterized in that: The method of using the culture device according to any one of claims 1 to 5 for culture comprises the following steps: (1) Preparation of nutrient solution: Dispense the prepared nutrient solution into the inner bottle of the culture bottle, add a quantity of culture solution just enough to cover the bottom of the inner bottle, then place in an incubator, seal, and sterilize; (2) Inoculation: Select seedlings and place them into the inner bottle of a culture bottle, then place the culture bottle into the culture space of an incubator and cover the incubator with a lid; (3) Cultivation: Place the incubator on a seesaw shaker, connect the air inlet of the incubator to the air inlet pipe, introduce CO2, start the shaker, and start culturing.

7. The seesaw liquid culture method according to claim 6, characterized in that: The culture conditions of step (3) are: culture temperature 23-27° C., light intensity 2400-3200 lx, light cycle 10-12 h, and culture for 50-70 days.

8. The seesaw liquid culture method according to claim 6, characterized in that: The rocking frequency of the seesaw shaker is 40-70 times / min.

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