CN107047265B - Circulating system for culturing high-abundance isotope carbon and nitrogen double-labeled plant sample - Google Patents

Circulating system for culturing high-abundance isotope carbon and nitrogen double-labeled plant sample Download PDF

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CN107047265B
CN107047265B CN201611267319.6A CN201611267319A CN107047265B CN 107047265 B CN107047265 B CN 107047265B CN 201611267319 A CN201611267319 A CN 201611267319A CN 107047265 B CN107047265 B CN 107047265B
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transparent cover
culture
circulating system
nutrient solution
carbon
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CN107047265A (en
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艾超
周卫
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Institute of Agricultural Resources and Regional Planning of CAAS
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Institute of Agricultural Resources and Regional Planning of CAAS
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G31/00—Soilless cultivation, e.g. hydroponics
    • A01G31/02—Special apparatus therefor
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00—Botany in general
    • A01G7/02—Treatment of plants with carbon dioxide
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
    • Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures

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  • Environmental Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biodiversity & Conservation Biology (AREA)
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Abstract

The invention relates to the technical field of isotope labeling, in particular to a method for culturing high-abundance isotope carbon,The circulating system of the nitrogen double-labeled plant sample comprises a closed culture chamber and CO2The device comprises an injector, a gas balance device, a nutrient solution injector and a nutrient solution circulating system; the sealed culture chamber comprises a transparent cover and a base, wherein a semiconductor condensation and dehumidification device is arranged in the transparent cover, and the transparent cover is connected with CO2An injector and a gas balancing device; the culture basin, the liquid storage chamber, the outward discharge peristaltic pump, the ultraviolet sterilization device, the circulating peristaltic pump and the converging pool are sequentially connected to form a circulating loop of the nutrient solution circulating system; the converging pool is connected with a semiconductor condensation dehumidifying device and a nutrient solution injector; the culture basin and the liquid storage chamber are arranged in the base, the top of the culture basin is communicated with the transparent cover, and quartz sand is arranged in the culture basin. The invention can simultaneously mark carbon and nitrogen isotopes and realize the recycling of the isotopes, and has no common isotope12C and14n pollution, and obtaining high abundance carbon and nitrogen isotope double-labeled plants.

Description

Circulating system for culturing high-abundance isotope carbon and nitrogen double-labeled plant sample
Technical Field
The invention relates to the technical field of isotope labeling, in particular to a circulating system for culturing a high-abundance isotope carbon and nitrogen double-labeled plant sample.
Background
In recent years, the circulation process of carbon and nitrogen elements contained in crop straws in soil has become one of research hotspots in plant nutrition and soil science. The isotope tracing technology is a key technology for researching the decomposition and conversion process of crop straws in soil, and can effectively reveal the release rule of straw elements and the biological effectiveness of organic nutrients. Stabilized isotope of nitrogen (15N) applying the marked crop straw into the soil, and determining the different components of the soil15The change of the N abundance can accurately calculate the straw15The amount of N nutrients transferred to soil, plants, and the proportion lost by gas, leaching, etc. Similarly, using stable isotopic carbons (C:)13C) Tracing, combining with modern molecular biology methods, a series of stable isotope probe technologies (SIP) are created to study and describe the decomposition direction of straw carbon and the biological process of synthesizing biomacromolecules through biochemical action, thereby further disclosing the microbiological mechanism of straw decomposition. However, the carbon and nitrogen conversion of the straws is an interconnected cyclic process, the carbon and the nitrogen conversion of the straws and the straw occur simultaneously, and the conversion mechanism is closely connected and inseparable. Therefore, the basis and the premise for researching the carbon and nitrogen conversion process of the straws are to obtain high-abundance isotopic carbon and nitrogen double-labeled plant samples.
The existing plant isotope labeling technology mainly marks single elements. Chinese invention patents CN200610019742.4, CN201310513820.6 and utility model patent cn201420736248.x all disclose a carbon dioxide isotope (c: (c) (r))13C-CO2) The marking device takes the plants cultured in the soil as the marking objects, and does not involve the simultaneous marking of two isotopes; although the invention patent CN201410342182.0 discloses an isotope stalk double-label tracing method, the method mainly utilizes13C-glucose and15the N-urea is directly injected into the plant body and is mainly used for researching root system secretion, the marking method does not belong to the normal assimilation category of the plant, and the marking method has great difference compared with the method that the plant assimilates carbon dioxide through normal photosynthesis and assimilates nitrogen through normal root system absorption.
Most isotope labeling methods use soil as culture medium, because the soil itself contains a large amount of common carbon atoms: (12C) By respiration of the microorganism, these carbon atoms are converted into12C-CO2The morphology is released in the air in large quantities and is absorbed and utilized by plants, resulting in marked plant samples13The C abundance decreases. In the process of researching the decomposition of crop straws, low-abundance isotope plant samples cannot trace carbon atoms at a molecular level (such as a DNA level); in addition, the method is carried out by taking soil as a culture medium15N-number of ordinary nitrogen atoms in the soil14N) is also absorbed by the plant, resulting in plant bodies15The N abundance is too low. Therefore, the selection of an appropriate culture mode is a prerequisite for obtaining a high-abundance isotope carbon and nitrogen double-labeled plant sample.
The prior plant isotope labeling technology is difficult to realize continuous and cyclic labeling of the whole plant growth period, which causes serious isotope waste and high labeling cost, for example,13C-CO2in the pulse marking process, when the closed space is opened at regular time, a large amount of the pulse marking is carried out13C-CO2Releasing; as in nutrient solutions under hydroponic conditions15The N nutrient can not be recycled, and a large amount of the old nutrient solution15N is wasted. The technical difficulty of continuous cycle labeling is the humidity of the air in the enclosed environmentThe isotope labeling of plants is failed due to the fact that the plant growth environment is worsened, the diseases such as mildew and the like are aggravated and the plant growth is blocked because of continuous enlargement; how to realize continuous cyclic labeling of stable isotope carbon and nitrogen is the key point for obtaining high-abundance double-labeled plant samples.
Currently, the art does not provide a systematic solution to the above problems. However, obtaining a high-abundance double-labeled plant sample is an essential material for researching the decomposition process and mechanism of carbon and nitrogen of plant residues. Therefore, a low-cost, high-efficiency and integrated circulatory system for culturing isotopic carbon-nitrogen double-labeled plants is needed.
Disclosure of Invention
Technical problem to be solved
The invention provides a circulating system for culturing a high-abundance isotope carbon and nitrogen double-labeled plant sample, which is used for solving the problems that the existing isotope labeling technology cannot realize the circulating culture of isotope carbon and nitrogen double-labeling and cannot obtain the high-abundance double-labeled plant sample.
(II) technical scheme
In order to solve the technical problems, the invention provides a circulating system for culturing a high-abundance isotope plant sample with double-labeled carbon and nitrogen, which comprises a closed culture chamber and CO2The device comprises an injector, a gas balance device, a nutrient solution injector and a nutrient solution circulating system; the closed culture chamber comprises a transparent cover on the upper part and a base on the lower part, the base is connected with the transparent cover, a semiconductor condensation dehumidifying device is arranged in the transparent cover, and the outer part of the transparent cover is hermetically connected with the CO2An injector and the gas balancing device; the nutrient solution circulating system comprises a culture basin, a liquid storage chamber, an external discharge peristaltic pump, an ultraviolet sterilization device, a circulating peristaltic pump and a confluence tank, wherein the culture basin, the liquid storage chamber, the external discharge peristaltic pump, the ultraviolet sterilization device, the circulating peristaltic pump and the confluence tank are sequentially connected to form a circulating loop; the merging pool is also connected with the semiconductor condensation dehumidifying device and the nutrient solution injector; cultivate the basin with stock solution room from the top down locates in proper order in the base, the top of cultivating the basin communicates the translucent cover, be equipped with in the cultivation basinThe quartz sand is filled with plant seeds which are accelerated to germinate.
The culture basin is communicated with the liquid storage chamber through a guide pipe, and a first floating ball liquid level switch is arranged in the liquid storage chamber.
The ultraviolet sterilization device comprises an ultraviolet sterilization chamber, an ultraviolet lamp and a second floating ball liquid level switch are arranged in the ultraviolet sterilization chamber, and the lower limit water level of the second floating ball liquid level switch is higher than the bottom of the ultraviolet sterilization chamber.
The semiconductor condensation and dehumidification device comprises a box body, and a radiator, a semiconductor refrigeration piece and a condenser which are connected in sequence are arranged in the box body.
The box body is internally provided with a fan, the fan is arranged at a position close to the radiator, and the side wall of the box body is provided with a vent hole.
The bottom of the box body is provided with a funnel close to the condenser, and the box body is sequentially communicated with the funnel and the confluence pool.
The transparent cover and the base are cylindrical parts with flanges at the opening ends, the flanges of the transparent cover and the flanges of the base are correspondingly connected, and a silica gel pad is arranged between the flanges of the transparent cover and the flanges of the base and used for achieving sealing connection of the transparent cover and the base.
The gas balance device comprises a gas balance tank, a piston transversely arranged is arranged in the gas balance tank, an opening communicated with the transparent cover is formed in the gas balance tank, and the opening is located on the lower portion of the piston.
Wherein the liquid storage area is hermetically connected with a waste liquid injector.
Wherein, CO is arranged in the transparent cover2An infrared sensor.
(III) advantageous effects
Compared with the prior art, the circulating system for culturing the high-abundance isotope carbon and nitrogen double-labeled plant sample has the following characteristics:
1. the invention cultures the high-abundance isotope carbon-nitrogen doubleThe circulation system for marking the plant sample realizes the simultaneous marking of carbon and nitrogen isotopes through the organic combination of the closed culture chamber, the gas balancing device and the nutrient solution circulation system, can realize the cyclic utilization of the nitrogen isotopes, is wholly sealed, and has no common12C and14and N pollutes the plants, so that the carbon and nitrogen isotopic abundance of the double-labeled plants obtained by culture is very high.
2. The nutrient solution circulating system is provided with an ultraviolet sterilization device which can ensure that the nutrient solution is in15N can be recycled after ultraviolet sterilization, does not change nitrogen form, does not affect plant absorption, and can be used for treating diabetes15The N-nutrient solution is secondarily utilized by plants, so that the isotope utilization rate is improved, and the marking cost is reduced;
3. the quartz sand as a culture medium is arranged in the culture area, so that the problem of soil respiration in the soil culture process can be solved12C-CO2And large amount of liquid absorption in hydroponics13C-CO2Thereby avoiding the influence on the reduction of the isotopic abundance of the plants in the processes of soil culture and water culture;
4. the problem of overhigh humidity of the culture room under a closed condition can be solved through the semiconductor condensation dehumidifying device in the transparent cover; by connecting the semiconductor condensation dehumidification device with the nutrient solution circulating system, condensed water can enter a culture area filled with quartz sand through the confluence tank, so that the water utilization rate can be improved, and the watering frequency can be reduced; the fan in the semiconductor condensing and dehumidifying device can promote the gas flow in the closed culture chamber and accelerate the CO2And the mixture is uniformly mixed, so that the utilization rate of crops is improved.
Drawings
FIG. 1 is a schematic diagram showing the composition of a circulatory system for culturing a carbon and nitrogen double-labeled plant sample with abundant isotopes provided by the invention;
FIG. 2 is a block diagram of a nutrient solution circulation system provided by the present invention;
FIG. 3 is a schematic diagram of a closed culture chamber according to the present invention;
in the figure, 1: sealing the culture chamber; 101: a transparent cover; 102: a base; 2: CO 22An injector; 3: gas balance devicePlacing; 301: a gas balancing tank; 302: a piston; 4: a nutrient solution injector; 5: a semiconductor condensation dehumidification device; 501: a box body; 502: a heat sink; 503: a semiconductor refrigeration sheet; 504: a condenser; 505: a fan; 6: a culture pot; 7: a liquid storage chamber; 8: an efflux peristaltic pump; 9: an ultraviolet sterilization device; 901: an ultraviolet sterilization chamber; 902: an ultraviolet lamp; 10: a circulating peristaltic pump; 11: a confluence tank; 12: quartz sand; 13: a plant; 14: a flow guide pipe; 15: a first float level switch; 16: a second float level switch; 17: a funnel; 18: a silica gel pad; 19: a waste liquid injector; 20: CO 22An infrared sensor; 21: and (4) a rubber plug.
Detailed Description
To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. The above description is only a preferred embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by using the contents of the present specification and the accompanying drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The invention provides a circulating system for culturing a high-abundance isotope carbon and nitrogen double-labeled plant sample, which is used for solving the problems that the existing isotope labeling technology cannot realize the circulating culture of isotope carbon and nitrogen double-labeling and cannot obtain the high-abundance double-labeled plant sample.
As shown in figures 1-3, the embodiment of the invention provides a circulating system for culturing a high-abundance isotope plant sample with double-labeled carbon and nitrogen, which comprises a closed culture chamber 1 and CO2Injector 2, gas balance device 3, campA nutrient solution injector 4 and a nutrient solution circulating system; the closed culture chamber 1 comprises an upper transparent cover 101 and a lower base 102, the base 102 is connected with the transparent cover 101 to form a completely closed culture chamber 1, a semiconductor condensation dehumidifying device 5 is arranged in the transparent cover 101, the semiconductor condensation dehumidifying device can be used for dehumidifying the inside of the closed culture chamber 1 at intervals in the culture process of plant samples, and the transparent cover 101 is externally connected with CO in a sealing mode2Injector 2 and gas balance 3, CO2The syringe 2 can be injected into the closed culture chamber 113C-CO2Thereby providing the plants with the desired marker13C-CO2(ii) a The transparent cover 101 is connected to an external gas balance device 3 through a pipe, and can be continuously fed into the closed culture chamber 113C-CO2The atmospheric pressure in the rear sealed culture chamber 1 is kept normal, and when the temperature of the sealed culture chamber 1 is reduced and the volume of the gas is reduced, the gas in the gas balance device 3 can flow back to the sealed culture chamber 1 in time, so that the gas can flow back into the sealed culture chamber 113C-CO2Can be fully utilized. In addition, the nutrient solution circulating system comprises a culture basin 6, a liquid storage chamber 7, an efflux peristaltic pump 8, an ultraviolet sterilization device 9, a circulating peristaltic pump 10 and a confluence tank 11, wherein the culture basin 6, the liquid storage chamber 7, the efflux peristaltic pump 8, the ultraviolet sterilization device 9, the circulating peristaltic pump 10 and the confluence tank 11 are sequentially connected to form a circulating loop; the confluence tank 11 is also connected with a semiconductor condensation and dehumidification device 5 and a nutrient solution injector 4; cultivate basin 6 and stock solution room 7 from the top down and locate in proper order in base 102, cultivate the top intercommunication translucent cover 101 of basin 6, be equipped with quartz sand 12 in cultivating basin 6, put plant 13 in the quartz sand 12, set up as culture medium quartz sand in cultivating basin 6, can solve soil and cultivate in-process soil respiration and produce12C-CO2And large amount of liquid absorption in hydroponics13C-CO2Thereby avoiding the influence on the reduction of the isotopic abundance of the plants in the processes of soil culture and water culture; when the plant 13 is illuminated through the transparent cover 101, the leaves of the plant 13 absorb CO by photosynthesis2The injector 2 is injected into the sealed culture chamber 113C-CO2Thereby realizing the labeling of the isotopic carbon; the root system absorption of the plant 13 flows into the culture pot 6 from the confluence tank 11Is/are as follows15N-nutrient solution, thereby realizing the labeling of isotope nitrogen; in the nutrient solution circulation system, the nutrient solution injector 4 can inject new nutrient solution into the confluence tank 11, thereby providing the plants with the new nutrient solution15N；15The N-nutrient solution enters the culture pot 6 from the confluence tank 11, and excessive N-nutrient solution enters the culture pot 615The N-nutrient solution flows into the liquid storage chamber 7 when the liquid storage chamber 7 is filled with the N-nutrient solution15When the N-nutrient solution is too much,15the N-nutrient solution enters an ultraviolet sterilization device 9 for sterilization under the action of an external peristaltic pump 8, and the sterilized N-nutrient solution15The N-nutrient solution is converged into the converging pool 11 again under the action of the circulating peristaltic pump 10, the problem that the nutrient solution cannot be frequently replaced under the closed condition can be solved through the nutrient solution circulating system, and the ultraviolet sterilization device 9 is arranged in the nutrient solution circulating system to ensure that the nutrient solution in the nutrient solution can be sterilized15N can be recycled after ultraviolet sterilization, does not change nitrogen form, does not affect plant absorption, and can be used for treating various diseases15The N-nutrient solution is secondarily utilized by plants, so that the utilization rate of isotopes is improved, and the marking cost is reduced; the semiconductor condensation dehumidifying device utilizes the Peltier effect of the semiconductor refrigerating sheet, partial water in the air can be condensed into water, the condensed water is converged into the converging pool 11 through a pipeline, the condensed water enters the culture basin 6 through the converging pool 11, the water-steam virtuous cycle in the sealed culture chamber 1 is realized, the air humidity in the sealed culture chamber 1 can be reduced, the condensed water can be reutilized by plants, the water utilization rate is improved, the watering frequency in the plant culture process is reduced, the dehumidifying process of the semiconductor condensation dehumidifying device belongs to physical dehumidification, other harmful chemical substances are not introduced, the size is small, the power consumption is low, the heat dissipation is small, and the semiconductor condensation dehumidifying device can continuously work.
The circulating system for culturing the high-abundance isotope carbon-nitrogen double-labeled plant sample realizes the simultaneous labeling of carbon and nitrogen isotopes and the cyclic utilization of the carbon and nitrogen isotopes through the organic combination of the closed culture chamber, the gas balancing device and the nutrient solution circulating system, is sealed as a whole, and has no common isotope12C and14and due to N pollution, the abundance of carbon and nitrogen isotopes of the double-labeled plants obtained by culture is high. The circulating culture system has high efficiency, easy operation and low cost; the whole of the plantThe marking process is completely closed, continuous marking can be realized, and prevention of13C-CO2The leakage of the water is prevented,15the N-nutrient solution can be recycled; the semiconductor condensation dehumidifying device can strictly control humidity, promote plant growth and reduce marking cost.
In this embodiment, as shown in fig. 2, the culture pot 6 is communicated with the liquid storage chamber 7 through the flow guide tube 14, and the liquid storage chamber 7 is provided with a first float level switch 15. Specifically, the bottom of the culture basin 6 is connected with the top of the liquid storage chamber 7 through a guide pipe 14 with the diameter of 1cm, and liquid in the quartz sand 12 permeates to the bottom of the culture basin 6 and enters the liquid storage chamber 7 through the guide pipe 14; the volume of the liquid storage chamber 7 is 1.5L, a small floating ball liquid level switch, namely a first floating ball liquid level switch 15 is arranged in the liquid storage chamber, and the volume of a space between the upper limit water level and the lower limit water level of the liquid storage chamber is 1L; the liquid storage chamber 7 is connected with an external ultraviolet sterilization device 9 through a drain pipe, and an external peristaltic pump 8 is additionally arranged on the drain pipe; when the liquid in the liquid storage chamber 7 rises to the upper limit water level, the first floating ball liquid level switch 15 starts the outer discharge peristaltic pump 8 to work, the outer discharge peristaltic pump 8 sets the flow rate to be 50ml/min, the liquid in the liquid storage chamber 7 is pumped into the ultraviolet sterilization device 9, and when the liquid in the liquid storage chamber 7 falls to the lower limit water level, the first floating ball liquid level switch 15 closes the outer discharge peristaltic pump 8.
In this embodiment, the ultraviolet sterilization apparatus 9 includes an ultraviolet sterilization chamber 901, an ultraviolet lamp 902 and a second float level switch 16 are disposed in the ultraviolet sterilization chamber 901, and a lower limit water level b of the second float level switch 16 is higher than the bottom of the ultraviolet sterilization chamber 901. Specifically, the ultraviolet sterilization chamber 901 is a darkroom, and ultraviolet lamps 902 are installed therein, it is understood that a plurality of ultraviolet lamps 902 may be provided, in this embodiment, 2 ultraviolet lamps 902 are additionally installed in the ultraviolet sterilization chamber 901, and the 253.7nm ultraviolet intensity radiated by the ultraviolet lamps is not lower than 70uW/cm 2; a small float level switch, namely a second float level switch 16, is arranged in the ultraviolet sterilization chamber 901, the volume of a space between the upper limit water level a and the lower limit water level b of the second float level switch 16 is 0.5L, and the volume of a space between the lower limit water level b of the second float level switch 16 and the bottom of the ultraviolet sterilization chamber 902 is 0.5L; when the liquid in the ultraviolet sterilization chamber rises to the upper limit water level a, the second float level switch 16 starts the circulating peristaltic pump 10 to work, so as to lead the liquid in the ultraviolet sterilization chamber 901 to flow15N-nutrientThe nutrient solution is pumped to a confluence tank 11 and finally returns to a culture basin 6 filled with quartz sand 12 culture medium, and when the liquid in the ultraviolet sterilization chamber 901 falls to a lower limit water level b, a second floating ball liquid level switch 16 closes the circulating peristaltic pump 10; a space of 0.5L is left between the lower limit water level b of the second float level switch 16 and the bottom of the ultraviolet sterilization chamber 901, so that liquid in the ultraviolet sterilization chamber 901 can be always in a sterilization state.
In this embodiment, the semiconductor condensing and dehumidifying device 5 includes a box 501, and a heat sink 502, a semiconductor cooling plate 503 and a condenser 504 which are connected in sequence are disposed in the box 501.
In this embodiment, a fan 505 is further disposed in the box 501, the fan 505 is mounted at a position close to the heat sink 502, and a vent hole is formed in a side wall of the box 501.
In this embodiment, the bottom of the box 501 is provided with a funnel 17 near the condenser 504, and the box 501 is communicated with the funnel 17 and the merging tank 11 in sequence.
The side close to the condenser 504 absorbs heat and refrigerates through the Peltier effect of the semiconductor refrigerating sheet 503, the side close to the radiator 502 releases heat, when moist air passes through the condenser 504, part of water in the air is condensed into water, the condensed water flows into the converging pool 11 through the funnel 17 and returns to the culture basin 6 through the converging pool 11, the fan 505 is arranged at one side of the radiator 502, and because the side wall of the box body 501 is provided with the vent hole, the air flow in the closed culture chamber 1 can be accelerated under the action of the fan 505, so that the air humidity in the closed culture chamber 1 is rapidly reduced; in addition, the fan 505 accelerates the gas flow in the sealed cultivation room 1 to promote the plant pairing13C-CO2The assimilation and utilization of (2) and the improvement of plant body13C abundance.
The maximum dehumidification capacity of the semiconductor condensation dehumidification device 5 is 500ml/24h, and the working time is set as follows: the semiconductor condensation dehumidifying device 5 works for 1 hour every 3 hours; after continuous marking, the air humidity of the closed culture chamber 1 reaches 100%, the fan is started to drive the air in the closed culture chamber 1 to flow, the humid air passes through the pipeline in the condenser 504, part of water in the air is condensed into water by using the Peltier effect of the semiconductor refrigerating sheet, and the condensed water returns to the culture basin 6 filled with the quartz sand 12 matrix through the funnel 17 and the drain pipe.
In this embodiment, as shown in fig. 3, in order to facilitate the connection between the transparent cover and the base, to form a sealed culture chamber 1 and to facilitate the detachment of the sealed culture chamber 1, a culture pot 6 and a liquid storage chamber 7 are conveniently placed, and after the plant 13 is cultured, the plant 13 is conveniently taken out, the transparent cover 101 and the base 102 are both cylindrical members with flanges at the opening ends, the flange of the transparent cover 101 corresponds to the flange of the base 102 and is connected, and a silica gel pad 18 is arranged between the flange of the transparent cover 101 and the flange of the base 102, so that the sealing performance between the flange of the transparent cover 101 and the flange of the base 102 can be ensured, the flange of the transparent cover 101 and the flange of the base 102 are both provided with a plurality of screw holes, and the bolts pass through the screw holes, so that the flange of the transparent cover 101 and the flange of the base 102 are connected.
Before the connection by the bolt, the culture pot 6 is placed in the base 102, and the outer diameter of the culture pot 6 is equal to the inner diameter of the base 6; putting quartz sand 12 into the culture pot 6, sterilizing the quartz sand 12 at 400 ℃ for 4 hours before use, pouring 0.5 times of Hoagland nutrient solution after cooling to saturate the water content of the quartz sand, and after adding the quartz sand 12, keeping the distance between the surface layer of the quartz sand and the upper edge of the culture pot 6 at 3 cm; the plant seeds which are accelerated to germinate are added into the quartz sand 12, the burying depth is 2cm, the seeds are soaked in 95% ethanol for 3 minutes before being accelerated to germinate, then soaked in 5% sodium hypochlorite for 8 minutes, and then washed with sterilized water for 3 times.
In this embodiment, the gas balance device 3 includes a gas balance tank 301, a piston 302 transversely disposed in the gas balance tank 301, and an opening communicating with the transparent cover 101 is disposed on the gas balance tank 301 and located at a lower portion of the piston 302.
The closed culture chamber 1 is externally connected with a gas balance device 3 through a pipeline, a piston 302 capable of freely moving up and down is arranged in a gas balance tank 301, and a guide pipe inlet is positioned below the piston 302; the volume of the gas balance tank 301 is more than 2L; while continuously adding into the closed culture chamber 113C-CO2Then, the internal gas pressure is increased, so that the excessive gas in the closed culture chamber 1 moves into the gas balance device 3, and the piston 302 moves upwards; on the contrary, when the culture chamber 1 is closedTemperature reduction or plant absorption of13C-CO2The piston 302 moves downward due to the decrease in the air pressure caused by the factors, and the air in the air balance device 3 moves toward the closed culture chamber 1, thereby maintaining the normal atmospheric pressure in the closed culture chamber 1.
In this embodiment, the liquid storage chamber 7 is hermetically connected with a waste liquid injector 19, and in the process of recycling the nutrient solution, the nutrient solution is recycled15After the N nutrient is gradually absorbed by the plants 13, the waste nutrient solution is extracted from the liquid storage chamber 7 through a waste liquid injector 19 connected outside the liquid storage chamber 7, and new nutrient solution is added into a culture basin 6 filled with quartz sand 12 matrix through a nutrient solution injector 4 connected with the confluence tank 11.
In this embodiment, CO is provided in the transparent cover 1012An infrared sensor 20. After the transparent cover 101 and the base 102 are sealed, CO of the three-way valve is used2The syringe 20 is quantitatively injected into the closed culture chamber 113C-CO2By means of CO2The infrared sensor 20 detects CO in the closed culture chamber 12Concentration of (2), CO in the sealed culture chamber 12The concentration is maintained at 300 ppm-600 ppm to ensure that plant 13 is not luxuriantly absorbed13C-CO2(ii) a Wherein, CO2The probe of the infrared sensor 20 is located inside the closed culture chamber 1, CO2The display screen of the infrared sensor 20 is positioned outside the closed culture chamber 1, the connecting line of the probe and the display screen passes through the transparent cover 1 through the rubber plug 21, CO2The detection range of the infrared sensor 20 is 1-2000ppm, and the accuracy is 1%. In addition, the transparent cover 101 can be made of transparent organic glass, and the base 102 can be made of PVC.
In this embodiment, the confluence tank 11 is disposed inside the transparent cover 101, and the connections of the circulation systems are all connected through pipes, such as: the nutrient solution circulating system is connected through a pipeline, the semiconductor condensation and dehumidification device 5 is connected with the confluence tank 11 through a pipeline, and the transparent cover 101 is connected with the gas balance device 3 through a pipeline. In order to strictly ensure the tightness of the closed culture chamber 1, all syringes and pipes are connected to the internal pipes of the closed culture chamber 1 through the rubber stopper 21 by means of needles.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims (7)

1. A circulating system for culturing a high-abundance isotope carbon and nitrogen double-labeled plant sample is characterized by comprising a closed culture chamber and CO2The device comprises an injector, a gas balance device, a nutrient solution injector and a nutrient solution circulating system; the closed culture room comprises a transparent cover on the upper part and a base on the lower part, the transparent cover and the base are cylindrical parts with flanges at the opening ends, the flanges of the transparent cover and the flanges of the base are correspondingly connected, a silica gel pad is arranged between the flanges of the transparent cover and the flanges of the base to realize the sealing connection of the transparent cover and the base, a semiconductor condensation dehumidifying device is arranged in the transparent cover, and the outer sealing connection of the transparent cover is realized for CO2An injector and the gas balancing device; the nutrient solution circulating system comprises a culture basin, a liquid storage chamber, an external discharge peristaltic pump, an ultraviolet sterilization device, a circulating peristaltic pump and a confluence tank, wherein the culture basin, the liquid storage chamber, the external discharge peristaltic pump, the ultraviolet sterilization device, the circulating peristaltic pump and the confluence tank are sequentially connected to form a circulating loop; the confluence tank is also connected with the semiconductor condensation dehumidifying device and the nutrient solution injector; the culture pot and the liquid storage chamber are sequentially arranged in the base from top to bottom, the top of the culture pot is communicated with the transparent cover, quartz sand is arranged in the culture pot, and plant seeds which are accelerated to germinate are placed in the quartz sand; the ultraviolet sterilization device comprises an ultraviolet sterilization chamber, an ultraviolet lamp and a second floating ball liquid level switch are arranged in the ultraviolet sterilization chamber, and the lower limit water level of the second floating ball liquid level switch is higher than the bottom of the ultraviolet sterilization chamber; the gas balance device comprises a gas balance tank, a piston transversely arranged is arranged in the gas balance tank, an opening communicated with the transparent cover is formed in the gas balance tank, and the opening is located on the lower portion of the piston.
2. The circulating system for culturing the carbon-nitrogen double-labeled plant sample with high abundance isotope according to claim 1, wherein the culture pot is communicated with the liquid storage chamber through a flow guide pipe, and a first float level switch is arranged in the liquid storage chamber.
3. The circulating system for culturing the high-abundance isotope carbon and nitrogen double-labeled plant sample as claimed in claim 1, wherein the semiconductor condensing and dehumidifying device comprises a box body, and a radiator, a semiconductor refrigerating sheet and a condenser are sequentially arranged in the box body.
4. The circulating system for culturing the carbon and nitrogen double-labeled plant sample with the abundant isotope as claimed in claim 3, wherein a fan is further arranged in the box body, the fan is installed at a position close to the heat sink, and a vent hole is formed in a side wall of the box body.
5. The circulating system for culturing the high-abundance isotope carbon-nitrogen double-labeled plant sample as claimed in claim 3, wherein the bottom of the box body is provided with a funnel close to the condenser, the top end of the funnel is communicated with the box body, and the bottom end of the funnel is communicated with the confluence pool.
6. The circulating system for culturing the high-abundance isotope carbon and nitrogen double-labeled plant sample as claimed in claim 1, wherein the reservoir is hermetically connected with a waste liquid injector.
7. The circulating system for culturing the high-abundance isotope carbon-nitrogen double-labeled plant sample as claimed in claim 1, wherein CO is arranged in the transparent cover2An infrared sensor.
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KR102383609B1 (en) * 2019-11-27 2022-04-06 주식회사 미드바르 Plant growing system using water extracted from the air
BR112022021723A2 (en) * 2020-05-01 2022-12-06 Pivot Bio Inc PLANT CULTIVATION SYSTEM, METHODS FOR DETECTING NITROGEN INCORPORATION IN A PLANT AND FOR IDENTIFICATION OF A NITROGEN FIXING BACTERIAL STRAIN, GENETICALLY MODIFIED BACTERIA, AND COMPOSITION
CN114080924B (en) * 2021-11-12 2024-03-01 中南林业科技大学 Cultivation device and method for marking woody plant nitrogen stable isotope
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