WO2005058022A1

WO2005058022A1 - Combination-type plant sugarless tissue culture propagation device and method thereof

Info

Publication number: WO2005058022A1
Application number: PCT/CN2004/001315
Authority: WO
Inventors: Yongtai Zhang
Original assignee: Yongtai Zhang
Priority date: 2003-12-19
Filing date: 2004-11-19
Publication date: 2005-06-30

Abstract

The invention relates to an agriculture device and method thereof. The device comprises some culture containers combined into a honeycomb-type, which are made of a transparence material, and section surfaces of circumference sides are grade hexagon and airproof-type. There are nutrition liquid overflow equipments, nami lamps, and humidity control equipment in containers. C02 accommodate equipment are placed outside of containers and connected inside of containers via conduits. A venthole of a blower is connected into a loop conduit and a temperature control equipment. A combination of difference manners is adopted in plant culture. Manners involve a cycle gas disinfectant manner, a C02 gas equalization cycle accommodate manner, a gas cycle-type temperature control manner, a gas cycle-type humidity control manner, a plant nami Lamp irradiation manner, a cycle-type nutrition liquid overflow accommodate manner, a side-light irradiation rotation-type light compensation manner. The invention may settle proems that include contaminated rate high, plant growth hypogenesis, stunt and death and so on. It is improved observably in young plant quality. The invention may shorten culture time also.

Description

Combined plant sugar-free tissue culture rapid propagation device and method thereof

The invention relates to a plant sugar-free tissue culture rapid propagation device and a method thereof, in particular to a method using nanometer light, a method for circulating replenishment of nutrient solution, a method for reducing temperature and controlling humidity by gas circulation, and a side-light rotation type light. A self-rotating plant artificial cultivation device and a method thereof combined with compensation methods belong to the field of agricultural machinery. Background technique

Through artificial environment regulation, the environmental conditions for plant growth are improved to make the plants perform photosynthesis. Through good light and carbon source supplementation, the plants have the advantages of fast growth rate and survival rate. Co ₂ is used as a carbon source for plant growth. It is supplemented by good light and carbon sources and adjusted by artificial environment to improve the environmental conditions for plant growth, to make its plants perform photosynthesis, increase plant growth rate and change plant growth season. . At present, scientific and technological personnel are continuously adjusting the technical conditions and improving the technical design to further improve the plant survival rate, effectively improve plant varieties, and effectively save energy and occupy space.

In the prior art literature search of the plant sugar-free tissue culture rapid propagation device, it was found that the Chinese patent application number is 02221205. 1. The patent applicant is: Kunming Institute of Environmental Science, named: Combined Plant Tissue culture and rapid propagation device, the technology is described as follows: This device places culture containers in each layer of the culture rack, the air inlets of each culture container are connected with the air inlet pipe, the inlet of the air pipe is connected with the outlet of the disinfection container, and the disinfection container The inlet is connected to the outlet of the flow control board, and the inlet of the flow control board is connected to the carbon dioxide carbon source and the air pump respectively. The power supply control box that can control the illumination of the light source in each culture container is installed on the culture rack. Valves; air compression pumps are equipped with air filters in and out, and pulleys are installed at the bottom of the culture rack; the outlet of the culture container is equipped with an adjustable air outlet nut consisting of a gland, a filter membrane, and a connecting tube; one is connected to the bottom of the disinfection container Transparent U-shaped tube.

Due to the use of ordinary fluorescent lighting (light wave frequency) and factors from the top light source (fixed light source) for a long time, the above technology has a large difference between its working condition and natural ecology. At the same time, because of the humidification and temperature control of C0 ₂ Unsystematic supply, plant light Cooperative use cannot be fully exerted, and plant growth is constrained. The disadvantages of the above-mentioned device are the large footprint and the low survival rate of tissue culture plants. In addition, because of the unreasonable layout of the co ₂ outlet pipe, the co ₂ utilization efficiency is low.

After a literature search of the prior art on a method for rapid propagation of combined plant sugar-free tissue culture, it was found that: the patent application number is 98121867. 9, and the name is: A sugar-free box-type plant tissue culture method, the technology is described as : The present invention relates to a method for cultivating plant tissue. A culture container is used instead of a bottle as a culture container to facilitate the plant's full absorption and gas exchange of CO ₂ , and to provide a sugar-free + culture container + CO ₂ optimal cultivation method. Due to the use of a fixed light source, the tissue culture seedlings are not completely grown, and the CO ₂ distribution is uneven, the heat dissipation is poor, and the cost of tissue culture is high. The survival rate of tissue culture seedlings in the field planting process is low, and it is easy to form "glass seedlings" (that is, The growth environment has changed slightly, and the survival rate of the seedlings has dropped significantly). Summary of the invention

The purpose of the present invention is to overcome the shortcomings in the prior art and provide a combined plant sugar-free tissue culture rapid propagation device and method thereof. Under the premise of ensuring that the light and carbon source of sugar-free culture micro-propagation are fully satisfied, combined with the basic principles of botany, aerodynamics, optics, and physics, it overcomes the fragile growth and development of plant foliage, plant migration The problem of low survival rate of the plant is that the present invention can save the tissue culture space and energy to the maximum; and the gas circulation method is used to solve the problem of controlling the temperature and humidity of the tissue culture container during the sugar-free tissue culture process, which simplifies The prior art sugar-free tissue culture process.

The present invention is achieved through the following technical solutions. The combined plant sugar-free tissue culture rapid propagation device of the present invention includes: a culture container, a temperature control device, a nutrient solution diffusion device, a humidity control device, a nano-lamp, a co ₂ device, and a speed regulation device. The motor and fan are connected as follows: The culture container is made of transparent material, and its peripheral cross-sectional shape is equilateral hexagon, which is sealed. Several combinations of equilateral hexagon culture containers are arranged in a honeycomb shape. The nutrient solution diffusion device, nano lamp, humidity control device, and co ₂ device are arranged outside the culture container, and are connected to the culture container through pipes respectively. The speed regulating motor is arranged at the bottom of the culture container, and the speed regulating motor and the co ₂ in the culture container are arranged. The air diffuser is connected, the fan is arranged on the upper part of the culture container box, the fan inlet is connected with the culture container box, the fan outlet is connected with the circuit pipe, and the circuit pipe is connected with the temperature Controls are connected.

The culture container is made of transparent material, and its peripheral cross-sectional shape is an equilateral hexagon, which is sealed. Inside the equilateral hexagonal culture container, three sides are made of reflective glass, and the reflective glass sides are according to the six sides of the hexagon. They are arranged at intervals so that natural light outside the culture container can enter the inside of the culture container through a transparent material without reflective glass. In this way, the culture vessel contains both a reflective light source made of reflective glass and an external light source.

Several culture vessels can be combined and arranged in a honeycomb shape. The equilateral hexagonal box design can maximize the structural volume of the culture vessel and the structure is relatively firm. Multiple equilateral hexagonal culture vessel bodies are easier to maximize the use of space. At the same time, it is also conducive to the mutual transmission of light sources between culture vessels, which is convenient for large-scale, factory-based tissue culture management.

Several culture layers are set from the top to the bottom of the culture container. The culture layers are flat vertebral bodies, and the cross-section is ladder-shaped. Each culture layer is vertically connected from top to bottom by a co ₂ diffuser. .

Each culture layer is spirally connected from the vertical direction to form a nutrient solution diffusion device composed of a plurality of culture layers from top to bottom. The top of the nutrient solution diffusion device is connected to the nutrient solution inlet tube, and the bottom of the nutrient solution diffusion device is connected to the nutrition. The liquid outlet pipe is connected.

diffuser tube from top to bottom through co ₂ culture vessel, and means associated with the trickling nutrient solution, uniformly thick with an eye-shaped aperture on the C0 ₂ diffusing tube of the cabinets, C0 ₂ gas clouds by the C0 ₂ diffusing pipes The small holes enter the culture container, so that the CO ₂ gas in the culture container is evenly distributed.

Nano lamp disposed from top to bottom along the inner diameter of the casing, and the C0 ₂ diffusing pipe in a parallel shape, scattered around the co ₂ culture vessel is located within the tube, with co ₂ motion to the diffuser tube with the same speed. Nano lamps can uniformly emit red light with a wavelength of 640-660nm and blue light with a wavelength of 430-450nm.

The temperature control device is connected to the C0 ₂ inlet pipe, the C0 ₂ inlet pipe is connected to the C0 ₂ device with a carbon source, and the temperature control device is connected to the water mist outlet pipe through the C0 ₂ outlet pipe. It is connected to the humidity control device, which is connected to the water mist inlet pipe.

The fan is arranged on the upper part of the box of the culture container, the air inlet of the fan is connected with the box, the air outlet of the fan is connected with the return pipe, and the return pipe is connected with the temperature control device.

The working principle of the combined plant sugar-free tissue culture rapid propagation device of the present invention is as follows: The CO ₂ carbon source passes through the CO ₂ inlet pipe, enters the CO ₂ outlet pipe through the temperature control device, and the CO ₂ gas is in the outlet pipe After passing through the water mist inlet pipe, it is humidified to become a co ₂ gas rich in moisture (vegetative nutrient solution water). The CO ₂ gas rich in moisture enters the co ₂ diffuser in the culture container through the CO ₂ outlet pipe. Through the small pinhole-shaped air holes distributed on the surface of the co ₂ diffuser, the co ₂ gas floats from the bottom to the top, and evenly oozes out of the pin holes of the co ₂ outlet tube, so that the co ₂ gas in the culture container Evenly distributed.

The fan draws out the excessively high temperature co ₂ gas in the cabinet, and the higher temperature co ₂ gas enters the temperature control device through the loop pipe. The temperature control device cools down the high temperature co ₂ gas according to the set temperature value to make it cool. reached the standard temperature co ₂ gas while the temperature of the control means also co ₂ gas after cooling the gas concentration detection, and the gas concentration set value, the intake pipe from the automatic replenishment C0 ₂ C0 ₂ gas, by cooling, C0 ₂ gas and recycled to supplement the trachea co _2, and culturing the co ₂ outlet pipe into the container. This not only improves the efficiency of using CO ₂ gas, but also saves the cost of cooling the culture vessel.

While the CO ₂ gas is circulating, the nutrient solution required for plant growth flows into the nutrient solution diffusion device in the culture container continuously through the nutrient solution inlet pipe, and slowly flows down to the nutrient solution diffused flow along the nutrient solution diffused stream The bottom of the device flows out from the nutrient solution outlet pipe and flows into the nutrient solution inlet pipe again through external circulation.

In the present invention, a nano lamp is placed on the inner diameter of the culture container as a fixed light source in the culture container. Since the nano lamp installed on the CO ₂ outlet pipe can rotate with the bearing seat, it is used as a dynamic light source. According to the light wave theory, red light with a wavelength of 640-660nm can stimulate the chlorophyll photosynthesis ability, which is conducive to the accumulation of carbohydrates in plants. Blue light with a wavelength of 430-450IM can promote the accumulation of plant proteins and non-carbohydrates. The wavelength range of the present invention is determined according to the strongest region of the plant's chlorophyll absorption spectrum. This wavelength is most suitable for the growth of the plant and can maximize the photosynthesis efficiency of the plant. At the same time, the fixed light source installed in the culture container, with the slow rotation of the CO ₂ outlet tube, uses the inclination of the spiral nutrient solution diffuser and the wavelength difference of the light wave generated by the fixed light source to simulate the natural light irradiation of the plant in nature. Produces uninterrupted change of strong and weak light, so that the plant can grow like in the natural environment, the normal stomata opening and closing of the plant leaf surface can be fully calcined, and the plant has strong vitality. The invention overcomes the problem of fragile growth and development of chlorophyll spores and the photosynthetic phosphorylation synthase system of plants caused by the use of stationary top light irradiation in the original device, which makes the leaf surface stomata opening and closing of the plant in the culture container blocked, and The survival rate of transplantation is low. In addition, the plant can receive the light wave irradiation on both the leaf surface and the stem of the plant through the continuous change of side light waves in the culture container, and the photosynthesis of the leaves and the stem simultaneously, which accelerates the growth rate of the plant, thereby improving the plant's Survival regulation ability when transplanted into the natural environment.

Carbon dioxide concentration and light conditions are the two most important factors for photosynthesis in plants. The atmospheric carbon dioxide concentration is only 330 ppm. If expressed in volume, it is only 0.03% of the atmosphere. For every gram of glucose synthesized by a plant, the leaves must be from 2250. Sufficient (one gram of glucose) carbon dioxide can be absorbed evenly in liters of air, so carbon dioxide concentration often becomes a limiting factor for photosynthesis in plants. The effect of CO ₂ concentration on the photosynthetic rate of plants has both a "saturation point" and a "compensation point". Therefore, the efficiency of carbon dioxide gas absorption by the plants in the culture container is particularly important. Because the container of the original device is flat, its C0 ₂ inlet holes and C0 ₂ outlet holes are on the same plane and perpendicular. According to aerodynamic analysis, during the process of C0 ₂ gas from the inlet holes to the outlet holes, the periphery of the box gas recycle stream is generated due to death: ½, so that the C0 ₂ gas cabinets uneven distribution of gas chi pleiotropic effects area showed "olive" distribution in the housing. About 2/5 of the plants in the cabinet cannot be replenished with fresh CO ₂ gas, which seriously affects the growth and health of the plants. C0 ₂ gas according to the present invention, a temperature-controlled box into the circuit line, the control box through the humidity in the coolant pipe on the C0 ₂ gas humidification, C0 ₂ gas is humidified enters through the loop conduit built in the C0 ₂ outlet pipe culture vessel, and from C0 ₂ pinhole holes uniformly leaking pipe, so that every corner of C0 ₂ gas incubator evenly distributed within the container. Through the absorption and photosynthesis of the plants in the culture container, it is extracted by a fan placed outside the culture container. The temperature of the extracted CO ₂ gas is relatively high, and the high-temperature CO ₂ gas enters the temperature control box through the circuit pipe. after cooling, the circuit line C0 ₂ gas to obtain a carbon source supplemented, coupled to reflux wet tissue culture box, it will ensure the efficient use of co ₂ overcomes the gas "dead ends", but also to the temperature of the tissue culture tank Make effective adjustments.

The original device did not solve the design of circulating nutrient solution. The tissue culture substrate can only be generated using agar, perlite, sand, vermiculite and other carriers. The tissue culture efficiency is reduced, the tissue culture cost is high, and the tissue culture substrate must be subjected to 120 ° C. It can be used after autoclaving, which increases the probability of contamination of tissue culture plants. In addition, due to the small gap between the substrates and poor ventilation, it will cause root rot. If the pH value of the substrate is not properly controlled, the solid substrate will accumulate a lot of harmful components, which will greatly affect the growth of the plants. The invention adopts the nutrient solution flooding technology, and replaces the soil environment by artificially created crop root growth environment, so that the nutrient solution directly contacts the plant root system without using a substrate to fix the root Department of Tissue Culture. When the nutrient solution flows through the roots of the plant along the nutrient solution diffused spiral ladder, and the supply is circulated, the plant can obtain various nutrients required for growth from the nutrient solution. The method saves water, fertilizer, and labor, can also make the plant roots, stems, and leaves balanced and robust, does not pollute the environment, and is conducive to the application of large-scale and industrialized plant cultivation.

The combined plant artificial cultivation method of the present invention is to use a circulating gas disinfection method, a gas co ₂ carbon source balanced circulation supplement method, a gas circulation type temperature control method, a gas circulation type humidity control method, a plant nano lamp irradiation method, and a circulation type. Nutrient solution replenishment method and side light rotation type light compensation method are used in combination for plant artificial cultivation.

The following further details the methods of the present invention:

1.Circulating gas disinfection method

The existing process requires artificial alcohol wiping and disinfection on the inner wall of the culture container, which not only has high labor cost, but also has a “dead spot for disinfection”, and has a high probability of contamination of the culture container. 'The present invention makes full use of the multifunctional characteristics of the gas circulation channel. When sterilizing the culture container, ozone (0 ₃ ) gas is injected into the circulation channel for sterilization and disinfection, because the new ecological oxygen atoms contained in ozone have strong oxidation. Ability to directly penetrate the cell wall and combine with "unsaturated bonds" in the body to take bacterial life, it has a high sterilization efficiency. At the same time, ozone is a natural gas. It is taken from the air and reduced to oxygen by itself during the sterilization process and returned to the air. No residue left, no secondary pollution and side effects.

The ozone disinfection step of the present invention is −

① When the culture vessel is first cultured or finished, before the tissue culture plant is placed in the culture vessel, ozone gas with a concentration of 3 to 4 PPM is injected into the culture vessel through the gas circulation channel, and the gas flow rate is 5 to 15 L / min. , Disinfection time 5 minutes;

② After 5 minutes, the remaining ozone gas will be discharged through natural emptying, and the disinfection process of the culture container before tissue culture is completed.

Its bactericidal effect is shown in the following table:

Total number of bacteria before treatment Ozone density Action time Average bactericidal rate

(cfu-g ¹ ) (PPM) (min) (%)

3.2 X 10 ² 0.4-0.5 5 99.5 01315

The invention can quickly and thoroughly kill viruses and bacteria in the air and the culture container, completely eliminates the "dead-end dead spots" existing in the manual disinfection method of the prior art, not only has high sterilization efficiency, but also greatly reduces the disinfection cost of the culture container. Easier development of factory tissue culture technology.

2. Gas C0 ₂ carbon source equilibrium circulation supplement method

The concentration of carbon dioxide (C0 ₂ ) and light conditions are the two most important factors for photosynthesis in plants. The concentration of C0 ₂ in the air is only 330 ppm. If expressed in volume, it is only about 0.3% of air. Each gram of plant synthesis For glucose, plant leaves need to absorb evenly enough CO ₂ from 2250 liters of air as a carbon source for synthesizing one gram of glucose. Therefore, the CO ₂ concentration in the culture container becomes one of the determining factors for plant photosynthesis. As the original technical method of the culture container, the C0 ₂ air inlet hole and the C0 ₂ air outlet hole are on the same plane and vertical line. According to the aerodynamic analysis, the process of C0 ₂ gas from the air inlet hole to the air outlet hole around the culture container The dead end of gas circulation will be caused by the airflow, so that the C0 ₂ gas in the culture container is unevenly distributed, and the effective area of the gas is "olive" distributed in the culture container. About 2/5 of the plants in the culture container cannot Getting fresh CO ₂ gas supplementation will seriously affect the growth and health of tissue culture plants, resulting in a long tissue culture cycle and a low survival rate. In the original technical method, the CO ₂ gas has a low utilization rate and serious waste, so that the cost of the carbon (CO ₂ ) source rises.

The invention uses a gas circulation channel to make CO ₂ gas enter the culture container through countless gas outlets distributed on the channel, so that CO ₂ gas can be evenly distributed in every corner of the incubator, and overcomes the shortcomings of the "dead corner" of the gas circulation caused by the air flow. Based on the aerodynamic principle, an external power system is used to make the gas flow in the gas circulation channel. The automatic detection system detects the gas concentration in real time, monitors the saturation point and compensation point of CO ₂ concentration on the photosynthesis of plants, and controls it in time. Supply and recycle of co ₂ gas. The use efficiency of co ₂ gas is improved, and the cost of carbon (co ₂ ) source is reduced.

Because the present invention is a combination of multiple methods and uses a variety of advanced technologies in the tissue culture process, the photosynthesis rate of the plants in the culture container is much faster than the original method, that is, the plant production speed is much faster than the original method. . During the tissue culture period, the concentration of co _{2 in the} culture vessel is controlled to

100 (Tl500ppm, the number of gas ventilation of the culture vessel 3~10h- result of C0 ₂ gas circulation use, the supply of C0 ₂ within the culture vessel is sufficient to meet the needs of the plant.

3. Gas circulation temperature control method The internal temperature of the culture container is an element that affects the growth and development of the plant. The original technical method caused the temperature in the culture container to not be effectively reduced due to problems such as the manufacturing materials and structural layout of the culture container and the use of an external air conditioning system for the temperature control method. The growth and development of the plant, its temperature control method uses an external air-conditioning system of the culture container, because the external air-conditioning system measures the temperature of the outside of the culture container, and then adjusts the temperature of the outside air of the container. The internal temperature of the container is therefore insensitive to changes in the temperature of the culture container, low temperature control efficiency, and high power consumption costs, which cause the plant tissue culture operation cost to increase, and at the same time cause uneven temperature adjustment inside the culture container, which seriously affects the growth and development of the plant And plant survival.

The invention overcomes the shortcomings of the original method, and controls the temperature in the culture container through automatic temperature measurement and temperature adjustment of the circulating gas in the circulation channel by an external power system, so it is very sensitive to temperature adjustment in the culture container. When the temperature is detected to be too high, the gas in the circulation channel can be directly cooled, and the cooled gas can directly enter the culture container through the gas circulation channel, which can quickly cause the temperature change in the culture container to make it reach tissue culture technology. Required temperature. The invention improves the temperature control efficiency, reduces the temperature control cost, and can meet the ideal temperature for plant growth and development.

In the photoperiod, when the light or pure radiation light flux density at the boundary of the culture container is large and

At 35Wnf ² , the gas temperature (DIF) will change. Therefore, according to plant requirements, the temperature control of the plant during light and dark periods is as follows: 0 ~ 3 days, the temperature is controlled at 25 ° C / 15 ° C (+ 10DIF), 4 ~ 6 days, temperature control is 20 ° C / 20. C (0DIF), 7 ~ 15 days, temperature control is 15 ° C / 25 ° C (-10DIF) o

4. Gas circulation humidity control method

The original method uses an external air-conditioning system to cool down, so when the temperature of the outer wall of the culture container is lower than the temperature in the culture container, the water vapor in the culture container will condense on the wall and the top of the container, especially in large culture containers. Although the wall and top of the culture container are covered with steam condensate, the relative humidity in the container is difficult to reach more than 90%. This makes the air humidity in the culture container often unable to meet the needs of plant growth, and it is easy to cause the plants to lose water and wither, and the plant survival rate is low.

The invention overcomes the shortcomings of the original method. During the circulation of the gas in the culture vessel through the gas circulation channel, the humidity measurement and humidity adjustment are performed directly on the gas in the gas circulation channel. To keep the humidity in the culture container within the optimal humidity range to ensure the normal growth and development of the plant.

In order to ensure the seedling growth rate and rooting rate of the plant, the present invention applies the following measures to the humidity measurement and humidity conditioning of the air in the culture container: The external humidity control system uses water rich in plant nutrient solution to spray, The humidification of the co ₂ gas in the container can directly increase the humidity of the gas in the cultivation container. The plants can not only absorb the CO ₂ gas from the air, but also the roots, stems, and leaves can absorb nutrients from the water mist rich in nutrient solution. To make plants grow faster. Because the CO ₂ gas in the culture container is rich in plant nutrient solution, during the plant tissue culture cycle, the humidity is controlled to 90% to 100% on the 0th to 5th days, and the humidity is controlled to 75 to 85% on the 6th to 15th days. After 15 days, seedlings can be directly transplanted into the field. Tissue culture seedlings do not need to be cultivated during the boosting period and have strong adaptability.

5. Plant nano-light irradiation method

Plant leaves are the main organs for photosynthesis, and chloroplasts are important organelles for photosynthesis. According to the light wave theory, the light radiated by the sun to the ground has a wavelength of about 300 to 2600 nm, of which 380 to 760 nm is visible light. Scientific practice has proven that the most effective wavelength of visible light for photosynthesis in plants is in the range of 400-700 nm. In the original method, a fluorescent lamp was used as an illuminating light source in the cultivation container. The visible light wavelength emitted by the fluorescent lamp was 350 to 600 nm, which could not meet the optimal light wave requirements required by the plant.

The combined artificial plant cultivation method of the present invention analyzes the optical properties of chlorophyll from plants, according to the strongest region of the chlorophyll absorption spectrum: one in the red light portion with a wavelength of 64 {T660 nm, and the other in the blue light portion with a wavelength of 430 to 450 nm. The invention uses nanotechnology and uses a special nano lamp as a light source. The wavelength range of the light wave of the nano lamp is determined according to the strongest region of the plant's chlorophyll absorption spectrum. This wavelength is most suitable for plant growth and can maximize the photosynthesis efficiency of the plant. Good condition. The red light with a wavelength of 64 {T660nm emitted by the present invention can stimulate the photosynthetic capacity of chlorophyll and is beneficial to the accumulation of carbohydrates in plants. The blue light with a wavelength of 430 ^ 450nm can promote the accumulation of plant proteins and non-carbohydrates. .

The prior art method ignores the utilization rate of light energy, and the loss of light energy is large. The side-lighting system of the invention is beneficial to promoting the growth of plants and controlling the height of the plants under the condition of cost saving. In the culture container, the light intensity of the side light system is 5 times that of the vertical light system, and the light energy received by the plant leaves is more uniform than that of the vertical light system. N2004 / 001315 With shorter internodes and slightly more leaf numbers, plants can accept more spectral distribution. The present invention realizes side light irradiation, and by using the reflective light source in the culture container, the light energy is greatly saved, and about 50% of the light from the reflective light source is reflected upward, and the other 50% of the light is reflected downward, so that the The light intensity is increased by 1.8 times, saving 54% of light energy consumption, reducing the heat generation of light energy, and saving gas cooling costs by 75%.

According to the present invention, during the plant tissue culture period, the light intensity of the nano-lamp is 100 (T1500LX, through reflection light source, the light intensity in the culture container is 2000-3000LX, and the light time is 5 hours per day; 6-15 days, The light intensity of the nano lamp is 2500 ~ 3500LX, and the light intensity in the culture container is 500 (T7000LX, the light time is 15 hours per day, and the seedlings emerge after 15 days by reflecting the light source.

6.Circulation nutrient solution replenishment method

'The method of the prior art does not solve the design of circulating nutrient solution', and its tissue culture substrate can only be generated using agar, perlite, sand, vermiculite and other carriers, the tissue culture efficiency is reduced, the tissue culture cost is high, and the tissue culture substrate must be It can be used after high temperature sterilization at 12CTC, which increases the probability of contamination of tissue culture plants. In addition, due to the small gap between the substrates and poor ventilation, the root rot phenomenon of the plant will be caused. If the ra value of the substrate is not properly controlled, the solid substrate will accumulate a large number of harmful components, which will greatly affect the growth of the plant.

The combined plant artificial cultivation method of the present invention uses advanced "soilless cultivation technology" to replace the original culture medium quality environment by the artificially-created crop root growth environment, so that the plant nutrient solution directly contacts the plant root system in a diffuse state. The cyclic supply of nutrient solution to the plant root system, the plant obtains various nutrients required for growth from the nutrient solution. The plant root system is not affected by the quality of the culture medium, which accelerates the absorption of nutrients by the plant, and the plant root system can stretch freely . The strong root structure greatly improves the survival rate of the plant. This method not only overcomes the problems of slow root development and uneven plant growth in the original method, but also makes the plant's roots, stems, and leaves balanced and robust. Since the circulating supply of the nutrient solution is realized, the cost of the culture medium is reduced, and the surrounding environment is not polluted, which is conducive to the application of large-scale and factory tissue culture of plants.

The nutrient solution has a diffuse flow velocity of 5 mm / s, and a plant root immersion depth of 1 cm.

7.Side light rotation type light compensation method

As plants, especially angiosperms, their chloroplasts in the cell follow the direction of light and light. The light intensity shifts. Under strong light, the chloroplasts move parallel to the direction of the light source in the plant cell wall to avoid excessive heating. In low light, the chloroplast moves vertically in the plant cell wall with the direction of the light source, and absorbs light energy as much as possible. This is the survival law formed by plants after hundreds of millions of years of evolution in nature.

According to the above-mentioned natural survival theory of plants, the present invention causes the plants in the culture container to undergo a spatial change through external power. When the space between the plants and the spatial position of the light source in the culture container are relatively changed, light compensation for side illumination is formed, and This side light rotates around the culture container, and the wavelength of the light wave received by the plant will be relatively changed, thereby forming an uninterrupted change of the strong and weak light. The static light source in the culture container is changed to a relatively dynamic light source, which simulates the plant in nature. Receiving dynamic illumination from natural light sources, the leaf surface stomata of the plant are kept open according to the principle of photosynthesis, the chlorophyll photosynthetic capacity is fully calcined, and the vitality of the plant is strong, making the tissue culture plant more adaptable in the field transplanting process. The present invention overcomes the fragile growth and development of the plant chlorophyll and its photosynthetic phosphorylase system caused by the original method using a stationary light source, which hinders the opening and closing of the leaf surface stomata in the culture container and the survival of the plant in the field. The disadvantage of low rates.

In the external power system of the present invention, the rotation speed is determined according to the light intensity during the tissue culture period during the plant tissue culture period. 0 to 5 days, the rotation speed is 10 cycles / hour; with the increase of light intensity, the tissue culture period is 5 to 15 days, the rotation speed is 15 cycles / hour.

In addition, a reflective surface is used on the inside of the culture container of the present invention, so that the plants in the culture container can simultaneously receive the side light source and the reflected light source, so that different wavelengths of the central light source can be irradiated, which can maximize the utilization of light energy. The plant is illuminated by the changing side light waves in the culture container, so that the leaves and stems of the plant can be illuminated by the light waves, and the photosynthesis of the leaves and the stems is carried out at the same time, which accelerates the growth rate of the plant, thereby greatly improving the plant. The ability to adjust to survival in the natural environment avoids the generation of "glass seedlings".

The gas circulation channel of the present invention has multifunctional characteristics. The gas circulation channel is not only a control channel for a temperature control system and a humidity control system, but also a disinfection control channel. CO2 gas can be supplemented through the gas circulation channel, and the humidity and temperature of the culture container can be controlled by adjusting and controlling the gas temperature and humidity in the circulation channel. When the culture container tissue culture period ends, the original method requires manual disinfection of the culture container, the process is complicated, and labor costs The cost is high. Even so, there is still a high probability of pollution. When sterilizing the culture container, ozone gas can be injected into the circulation channel to sterilize and sterilize, which not only has high sterilization efficiency, but also greatly reduces the cost of sterilization.

The plant cultivated by the side-light rotation-type light compensation method of the present invention is significantly shorter than the tissue culture plant cultivated in the culture container adopted in the original method, but the plant weight is significantly increased, and the plant is robust, so the survival rate is high. The invention can be applied to the needs of tissue culture plants of various varieties. The application of the invention can improve the level of agricultural production technology in our country, reduce agricultural costs, increase farmers' income, guarantee the excellent quality of species, diversity of species, and improve the ecological environment. Is of great significance.

The invention has a great breakthrough in the traditional plant planting process, and solves the problems of high pollution rate, poor growth and development of plants, physiological morphological disorder, deformity, delayed growth or death in traditional tissue culture, which can significantly improve Seedling quality, shorten the cultivation period, improve labor productivity, and reduce production costs. The use of this technical rice is of great significance to improve the level of agricultural production technology, ensure the excellent quality of species, species diversity, improve the ecological environment, and increase farmers' income. BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 Schematic diagram of the invention (1)

Figure 2 Schematic diagram of the present invention (2)

Figure 3 Schematic diagram of the space layout of the present invention

As shown in FIGS. 1, 2, and 3, the combined plant sugar-free tissue culture rapid propagation device of the present invention includes: a culture container 1, a temperature control device 2, a nutrient solution diffusion device 3, a humidity control device 4, a nano lamp 5, C0 ₂ device 6, speed control motor 7, fan 8, C0 ₂ air diffuser 9, reflective glass 10, culture layer 11. The connection method is as follows: The culture container 1 is made of a transparent material, and its peripheral cross-sectional shape is an equilateral hexagon, which is sealed. The equilateral hexagon culture containers 1 are arranged in a honeycomb pattern, and nutrition is arranged in the culture container 1. The liquid diffusion device 3, the nano lamp 5, the humidity control device 4, and the C0 ₂ device 6 are installed in the culture container 1 and connected to the culture container 1 through pipes, respectively. The speed regulating motor 7 is provided at the bottom of the culture container 1 to adjust the speed. Motor 7 and training The C0 ₂ diffuser pipe 9 in the container is connected. The fan 8 is arranged on the upper part of the box of the culture container 1. The air inlet of the fan 8 is connected to the box of the culture container 1. The air outlet of the fan 8 is connected to the return pipe. The return pipe is connected to the temperature control. Device 4 is connected.

Inside the equilateral hexagonal culture container 1, three sides are provided with reflective glass 10, and the sides of the reflective glass 10 are arranged at intervals of the six sides of the hexagon.

Several culture layers 11 are arranged inside the culture container 1 from top to bottom. The culture layers 11 are flat circular vertebral bodies, and the cross section is a ladder shape. Each culture layer 11 is vertical from a C0 ₂ air diffuser 9 from top to bottom. Connected vertically.

Each culture layer 11 is spirally connected from a planar direction to form a nutrient solution diffusion device 3 composed of a plurality of culture layers 11 from top to bottom. The top of the nutrient solution diffusion device 3 is connected to a nutrient solution inlet pipe, and the nutrient solution diffuser 3 The bottom end is connected with the nutrient solution outlet pipe.

The CO ₂ diffuser 9 runs through the culture vessel 1 from top to bottom, and is connected to the nutrient solution diffusion device 3. The CO ₂ diffuser 9 is densely covered with pinhole-like holes.

The nano lamp 5 is arranged along the inner diameter of the culture container: from top to bottom, and is installed at six angular intervals of an equilateral hexagonal culture container. The nano lamp 5 and the C 0 ₂ air diffuser 9 are mounted on the C 0 ₂ air diffuser 9 It is parallel and located around the CO ₂ diffuser 9 in the culture container 1, and moves in the same direction and at the same speed as the CO ₂ diffuser 9. The red light with a wavelength of 640-660nm and the blue light with a wavelength of 430-450nm are emitted by the nano lamp 5.

The temperature control device 2 is connected to the C0 ₂ inlet pipe, the C0 ₂ inlet pipe is connected to the C0 ₂ device 6 with a carbon source, and the temperature control device 2 is connected to the water mist outlet pipe through the C0 ₂ outlet pipe. The mist outlet pipe is connected to the humidity control device 4, and the humidity control device 4 is connected to the water mist inlet pipe.

Examples and comparative tests of the combined plant artificial culture method of the present invention are as follows: Tissue culture types: Dioscorea seedlings

Cultivation method:

An equilateral hexagonal container made of a light-transmitting material + a reflective material installed along the inner space of the container + side light irradiation + C0 ₂ gas circulation cooling + nutrient solution (covering an area of 3.5 square meters).

Operation process:

This embodiment makes full use of the versatility of the CO ₂ gas circulation channel. At the time of tissue culture or tissue culture, before the tissue culture plant is placed in the culture container, ozone gas with a concentration of 3 to 4 PPM is injected into the culture container through a gas circulation channel, the gas flow rate is 5 to 15 L / min, and the disinfection time is 5 minutes. After 5 minutes, the remaining ozone gas is discharged through natural evacuation. During the sterilization process, the ozone gas is reduced to oxygen and returned to the air, leaving no residue, no secondary pollution and side effects. The disinfection method eliminates the "dead-end dead spots" caused by artificial disinfection, which not only has high sterilization efficiency, but also greatly reduces disinfection costs.

In this embodiment, the circulating nutrient solution for plant growth is culture medium. During the nutrient solution circulation cycle, the circulating nutrient solution is continuously sterilized by 12CTC, which solves the problem of plant growth and development caused by incomplete disinfection of the medium. The problem of contamination, this embodiment omits the process of "sterilizing first and then cultivating" the "culture substrate" of the tissue culture plant. On the basis of simplifying the tissue culture process and reducing the cost of the culture medium, the probability of contamination of the plant is greatly reduced.

In this embodiment, the principle of side-light dynamic irradiation is applied during the culture period. The light intensity of the nano lamp is 1000 to 1500 LX and the light time is 5 hours per day during 0 to 5 days. The light intensity of the nano lamp is 2500 to 6 to 15 days. 3500LX, the lighting time is 15 hours per day. Because the side light is used, and the light source is a dynamic and static combined light source, the light intensity can meet the photosynthesis needs of the plants in the culture container. The tissue culture period is 15 days. During the tissue culture period, the CO ₂ concentration in the culture container is controlled to be 1000 to 1500 ppm, and the nutrient solution diffuse flow rate is 5 mm / s. On day 0 ~ 5, the rotation speed of the culture layer driven by the external power system is 10 cycles / hour, the light intensity in the culture container is 2000 ~ 3000LX, the humidity control is 90% ~ 100%, and the temperature control is 23 ° C ~ 24 ° C . With the gradual growth of the plant, when the tissue culture period is 5 to 15 days, the light intensity in the culture container increases to 5000 to 7000 LX, the rotation speed is 15 cycles / hour, the light intensity in the culture container is 5000 to 7000 LX, and the humidity control is 75 to 85. %, Temperature control is 22 ° C ~ 25 ° C, root immersion depth of plant is lcm. After 15 days, the seedlings can be directly transplanted into the field, and the tissue culture seedlings do not need to be cultivated during the boosting period, and have strong adaptability. In this embodiment, the supply of CO ₂ gas, the humidity of the air, and the cooling of the culture vessel are achieved through an external power system. Therefore, the air circulation speed is adjustable, and the range of the controlled circulation speed is 3 to 10 hours for the gas exchange of the culture vessel— according to After the tissue culture test was performed on yam, the test results are compared as follows: Comparative Project Original Patent (02221205.1) This Example

Tissue culture period (days) 27 25

Lotus length (mm) 83 ± 15a 75 ± 10a

Fresh weight (mg) 750 ± 170a 800 ± 200a

Dry weight (rag) 74 ± 20a 80 ± 15a

Blade number 34 ± 0.6a 35 ± 0.3a

Leaf area (cm) 25 ± 5.4a 30 ± 6a

Number of transplanted plants 1505 3087

Number of viable plants 1083 3054

Survival rate (%) 72 98.9

Rooting radius (cm) 3 6.7

Cooling efficiency (%) 54 100

The above test results prove that the tissue culture period of this embodiment is short and the survival rate is high, and the cost of tissue culture is greatly reduced. The main reason for the decrease in cost is-

(1) Reduced raw material costs and labor costs. This embodiment completely replaces the original tissue culture substrate with the plant nutrient solution, which saves the cost of raw materials required for tissue culture. Since the disinfection process of the tissue culture substrate is omitted, the labor cost is also reduced accordingly.

(2) The consumption of electricity is reduced. The reflective material applied in the tissue culture container of this embodiment improves the light energy utilization rate by about 50%. Plant growth and development are accelerated, and the cultivation period is shortened by 409 ^ 60%, which reduces the lighting time and is the main reason for saving power consumption.

(3) The number of tissue culture plants increased. Compared with the prior art, in this embodiment, under the same cultivation area and the same planting density, the growth rate of the plant tissue culture is increased to 100%, and the seedling formation rate is also significantly improved.

(4) Plant loss is reduced. Compared with the prior art, this embodiment further reduces the influence of factors that cause the plant to be contaminated. In addition, with the cooperation of the automatic control system, it can produce high-quality tissue culture seedlings in a short tissue culture period.

Claims

1. A combined plant tissue culture and rapid propagation of sugar-free apparatus, comprising: a culture vessel (1), C0 ₂ (6), the fan (8), characterized in that, further comprising: a temperature control means (2), nutrients The liquid diffusion device (3), the humidity control device (4), the nanometer lamp (5), and the speed-regulating motor (7) are connected in the following manner: The culture container (1) is made of a transparent material and has a peripheral cross-sectional shape It is an equilateral hexagon, sealed, and several equilateral hexagonal culture vessels (1) are arranged in a honeycomb pattern. The culture vessel (1) is provided with a nutrient solution diffusion device (3), a nano lamp (5), and humidity. The control device (4) and ∞ ₂ device (6) are arranged in the culture container (1), and are connected to the culture container (1) through pipes, respectively, and the speed regulating motor (7) is arranged at the bottom of the culture container (1). The speed regulating motor (7) is connected to the C0 ₂ air diffuser (9) in the culture container (1), and the fan (8) is arranged in the ± part of the box of the culture container (1), and the air inlet of the fan (8) and the culture container (1) The box is connected, the air outlet of the fan (8) is connected with the circuit pipe, and the circuit pipe Temperature control means (4) is connected.

2. The combined plant sugar-free tissue culture rapid propagation device according to claim 1, characterized in that three sides of the culture container (1) are provided with reflective glass (10), and the sides of the reflective glass (10) are Arranged at six sides of the hexagon.

3. The combined plant sugar-free tissue culture rapid propagation device according to claim 1, characterized in that the culture container (1) is provided with a plurality of culture layers (11) from the top to the bottom, and the culture layers (11) It is a flat circular cone with a ladder-shaped cross section. Each culture layer (11) is vertically connected by a C0 ₂ air diffuser (9) from top to bottom.

4. The combined plant sugar-free tissue culture rapid propagation device according to claim 3, characterized in that each of said culture layers (11) is connected in a spiral shape from a planar direction to form a plurality of cultures from top to bottom The nutrient solution diffuser (3) is composed of a layer (11), the top of the nutrient solution diffuser (3) is connected to the nutrient solution inlet pipe, and the bottom of the nutrient solution diffuser (3) is connected to the nutrient solution outlet pipe.

5. The combined plant sugar-free tissue culture rapid propagation device according to claim 1 or 3, characterized in that the culture container (1) penetrates the C0 ₂ air diffuser (9), C0 ₂ powder from top to bottom The trachea (9) is connected to the nutrient solution diffuser (3), and the C0 ₂ diffuser (9) is uniformly and densely covered with pinhole-shaped holes.

6. The combined plant sugar-free tissue culture rapid propagation device according to claim 1, It is characterized in that the nano-lamps (5) are arranged from top to bottom along the inside diameter of the culture container (1), are installed at six angular intervals of an equilateral hexagonal culture container, and are installed in a CO ₂ diffuser ( nano lamp (5) and the C0 ₂ diffusing pipe (9) in a parallel shape, positioned culture vessel (C0 ₂ diffusing pipe (9)) within a surrounding, with the C0 ₂ diffusing pipe (9) with the same speed on 9) motion.

7. The combined plant sugar-free tissue culture rapid propagation device according to claim 6, wherein the nano-lamp (5) emits red light with a wavelength of 640-660 nm and blue light with a wavelength of 430-450 nm .

8. A modular plant according to claim 1, said sugar-free tissue culture and rapid propagation means, wherein said temperature control is connected to the intake pipe means C0 ₂ _(2), C0 2 with an intake pipe and a carbon source The C0 ₂ device (6) is connected, and the temperature control device (2) is connected to the water mist outlet pipe through the C0 ₂ outlet pipe, which is in a "bu" shape.

9. The combined plant sugar-free tissue culture rapid propagation device according to claim 8, wherein the water mist outlet pipe is connected to a humidity control device (4), and the humidity control device (4) It is also connected to the water mist inlet pipe.

10, a combined plant artificial culture method, characterized in that a circulating gas sterilization process, the carbon source gas C0 ₂ circulation complementary equalization method, the gas circulating a temperature control method, a control method for the gas circulating humidity, light plant nano The combination of the irradiation method, the circulating nutrient solution replenishment method, and the side-light rotation type light compensation method are used for plant artificial cultivation.

11. The combined plant artificial cultivation method according to claim 10, wherein the circulating gas sterilization method refers to injecting ozone gas into the circulation channel to sterilize and disinfect the culture container when sterilizing the culture container. The steps are:

12, a combined plant according to claim 10, wherein the artificial culture method, wherein said carbon source gas equilibrium C0 ₂ circulation complementary method is the application of air circulating passage so that C0 ₂ gas is distributed through the gas passage numerous The outlet enters the culture vessel, so that the CO ₂ gas can be uniform Distributed in all corners of the incubator, an external power system is used to form a gas flow in the gas circulation channel. The automatic detection system detects the gas concentration in real time, monitors the saturation point and compensation point of co ₂ concentration on photosynthesis of plants, and controls the co in time. ₂ Gas supply and recycling.

13. The combined plant artificial cultivation method according to claim 12, characterized in that during the tissue culture period, the concentration of CO _{2 in the} culture container is controlled to 1000 to 1500 ppm, and the number of gas exchanges of the culture container is 3 to 10 h - As a result of the use of C0 ₂ gas circulation mode, the supply of C0 ₂ within the culture vessel is sufficient to meet the needs of the plant.

14. The combined plant artificial cultivation method according to claim 10, wherein the gas circulation temperature control method is

When the temperature is detected to be too high, the gas in the circulation channel is directly cooled, and the cooled gas directly enters the culture container through the gas circulation channel, which rapidly causes the temperature change in the culture container to meet the technical requirements for tissue culture. Temperature, such as T: 0 ~ 3 days, temperature control: 25 ° C / 15 ° C (+ 10DIF), 4 ~ 6 days, temperature control: 20 ° C / 20 ° C (0DIF), 7 ~ 15 days, temperature control is 15. C / 25 ° C (-10DIF).

15. The combined plant artificial cultivation method according to claim 10, wherein the gas circulation type humidity control method refers to applying the following measures to the humidity measurement and humidity conditioning treatment of the air in the culture container: external type The humidity control system uses water rich in plant nutrient solution for spraying. By humidifying the CO ₂ gas entering the cultivation container, the humidity of the gas in the cultivation container can be directly increased. During the plant tissue culture cycle, on the 0th to 5th day, Humidity control is 90% ~ 100%, 6 ~ 15 days, humidity control is 75 ~ 85%, and after 15 days, the seedlings can emerge directly into the field for transplanting.

16. The combined plant artificial cultivation method according to claim 10, wherein the method for irradiating a plant with a nano-lamp means that the light intensity of the nano-lamp is within a range of 0 to 5 days during a plant tissue culture period. 1500LX, through the reflected light source, the illumination in the culture container is 2000-3000LX, the illumination time is 5 hours per day; 6 ~ 15 days, the light intensity of the nano lamp is 2500 ~ 3500LX, and the reflected light source, the illumination in the culture container is 5000 ~ 7000LX, the light time is 15 hours per day, and the seedlings emerge after 15 days.

17. The combined plant artificial cultivation method according to claim 10, wherein the circulating nutrient solution replenishment method refers to a crop root growth loop created artificially The original environment replaced the culture medium environment of the original method, so that the plant nutrient solution was in direct contact with the plant roots in a diffuse state. The diffuse flow rate of the nutrient solution was 5 mm / s, and the plant root immersion depth was 1 cm. The nutrient solution was used to supply the plant roots in a circulating manner. The liquid obtains various nutrients required for growth.

18. The combined plant artificial cultivation method according to claim 10, wherein the side-light rotation-type light compensation method refers to the use of external power to cause a spatial change in a plant in a culture container. The spatial distance changes relative to the spatial position of the illuminating light source in the culture container, forming a light compensation of the side light, and this side light rotates around the culture container, turning the static light source in the culture container into a relatively dynamic light source, simulating the plant in the natural world It receives dynamic illumination from natural light sources, and determines the rotation speed according to the light intensity during the tissue culture period. 0 to 5 days, the speed is 10 cycles / hour; with the increase of the light intensity, the tissue culture period is 5 to 15 days, and the rotation speed is 15 cycles / hour.

19. The combined plant artificial cultivation method according to claim 18, characterized in that, a reflective surface is used on the inside of the culture container, so that the plants in the culture container can simultaneously receive the side light source and the reflective light source, so that the middle light source Irradiation with different wavelengths can maximize the utilization of light energy.