CN115046968A - System and method for synchronously measuring photosynthetic rates of different tissues and organs of plant - Google Patents
System and method for synchronously measuring photosynthetic rates of different tissues and organs of plant Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/44—Sample treatment involving radiation, e.g. heat
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N2021/635—Photosynthetic material analysis, e.g. chrorophyll
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- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/25—Greenhouse technology, e.g. cooling systems therefor
Abstract
The invention discloses a system and a method for synchronously measuring photosynthetic rates of different tissues and organs of a plant, wherein the system comprises a control module, a control output end of the control module is connected with a control input end of a relay, two control output ends of the relay are respectively connected with control input ends of a first electromagnetic valve and a second electromagnetic valve, and a gas input end of the first electromagnetic valve is connected with a gas input end of the second electromagnetic valve 13 CO 2 The mist air supply is connected, the gas output end of first solenoid valve is connected with an input of flowmeter, the gas input end of second solenoid valve is connected with the air pump, the gas output end of second solenoid valve is connected with another input of flowmeter, the output of flowmeter communicates with the assimilation case through a plurality of pipeline, the assimilation incasement is provided with the plant that awaits measuring. What is needed isThe system has the advantages of wide tissue and organ, high measuring efficiency, consistent marking time and the like.
Description
Technical Field
The invention relates to the technical field of photosynthetic rate measurement methods and systems, in particular to a system and a method for synchronously measuring photosynthetic rates of different tissues and organs of a plant.
Background
It is generally accepted that carbon assimilation in plants depends primarily on leaf photosynthesis. However, some recent studies have found that chlorophyll-containing non-leaf organs such as pericarp, stalk, rice and wheat ear, etc. also have strong photosynthetic carbon assimilation ability (Chang et al, 2020; Simkin et al, 2020). The rice leaf sheath also contains chlorophyll, but its photosynthetic function is rarely studied. In fact, the photosynthetic function of the leaf sheath may be of great significance to the plant at seedling stage. Firstly, the volume of the leaf sheath of the rice plant in the seedling stage accounts for about 70% of the volume of the leaf, secondly, the early plant is small in size, the mutual shielding among individuals is small, the ratio of the light energy intercepted by the leaf sheath is high, and the leaf sheath is an important photosynthetic production tissue in the seedling stage of the rice. The traditional photosynthetic determination system is mainly used for determining the level of single leaves and population, and the lack of a high-flux in-situ determination tool for each organ of rice is one of the bottlenecks. Therefore, a rapid and accurate photosynthetic rate determination method for evaluating photosynthetic potential and contribution of each organ of rice is urgently needed at present.
The determination of the gas exchange rate is the most direct way to evaluate the photosynthetic rate of plants, and its main working principle is to seal all or part of the leaves in an assimilation box and to determine the CO in the gas entering and leaving the assimilation box by using an infrared gas analyzer (IRGA) 2 The photosynthetic rate was calculated from the difference in concentration of (c). In general, the plant growing environment is as followsThe environmental conditions in the assimilation box of the gas exchange determination system are different, and the blade needs a period of time to adapt to the new environment in the assimilation box (5-30min), which greatly limits the determination efficiency of the gas exchange determination system. In addition, the measurement time of the photosynthetic rate and the measurement of the leaf position in one day affect the measurement result.
Disclosure of Invention
The invention aims to solve the technical problem of how to provide a synchronous determination system for photosynthetic rate of different tissues and organs of plants, which has wide measurable tissues and organs, high determination efficiency and consistent marking time.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows: a system for synchronously measuring photosynthetic rates of different tissues and organs of a plant is characterized by comprising: the control output end of the control module is connected with the control input end of the relay, the two control output ends of the relay are respectively connected with the control input ends of the first electromagnetic valve and the second electromagnetic valve, and the gas input end of the first electromagnetic valve is connected with the control input end of the second electromagnetic valve 13 CO 2 The gas source of the mixed gas is connected, the gas output end of the first electromagnetic valve is connected with one input end of a flow meter, the gas input end of the second electromagnetic valve is connected with an air pump, the gas output end of the second electromagnetic valve is connected with the other input end of the flow meter, the output end of the flow meter is communicated with an assimilation box through a plurality of pipelines, a plant to be tested is arranged in the assimilation box, an air stirring device and an environmental parameter measuring sensor are arranged in the assimilation box, and the control input end of the air stirring device is connected with the control output end of the control module and used for acting under the control of the control module; the environment parameter measuring sensor is connected with the signal input end of the control module and used for transmitting measured data to the control module for processing; the artificial light source is positioned at the outer side of the assimilation box, is controlled by the control module and is used for providing illumination for the system; the gas treatment device is connected with the gas outlet of the assimilation box through a pipeline, and gas exhausted from the assimilation box is treated by the gas treatment device and then exhausted into the air.
The invention also discloses a method for synchronously measuring the photosynthetic rate of different tissues and organs of the plant, which uses the measuring system and is characterized by comprising the following steps:
selecting and planting test materials;
constructing and assembling a measuring system;
13 CO 2 determining the marking time;
13 CO 2 labelling experiments
13 C abundance determination;
and (6) analyzing the data.
The further technical proposal is that 13 CO 2 The method for determining the marking time comprises the following steps:
photosynthetic gas exchange rate was accomplished using LI-COR 6800, setting photosynthetic apparatus reference cell CO 2 The concentration is 400 mu mol -1 VPD is 1.2KPa, and the horizontal illumination intensity of leaves is 1500 mu mol m -2 s -1 (ii) a Recording photosynthetic gas exchange data after the variation amplitude of the net photosynthetic rate within 30s is lower than 5%; then the first solenoid valve and the second solenoid valve are controlled to switch the air inlet to 13 CO 2 Mixing gas, starting an automatic illumination control program of the photosynthetic apparatus, respectively marking different leaves for 20s, 40s, 60s, 120s, 180s, 240s, 300s, 600s, 1200s and 1800s, and automatically switching the illumination intensity to 0 mu mol m after the respective marking time is finished -2 s -1 And then cutting leaves in the assimilation box, photographing for calculating the area of the leaves, quickly placing the leaves in liquid nitrogen, drying at 80 ℃ to constant weight, and grinding a tissue sample to be detected after a powder state by using a ball mill.
The further technical proposal is that 13 CO 2 The method for labeling the assay comprises the steps of:
placing prepared test material in assimilation box under the treatment of light shielding and non-light shielding of leaf sheath, supplying air for 30min with air pump, allowing the plant body to adapt to the environment in assimilation box, controlling the second solenoid valve to close, the first solenoid valve to open, and controlling the flow rateThe meter control comprises 13 CO 2 The mixed gas enters an assimilation box at the speed of 20L/min, and the gas discharged from the assimilation box absorbs waste gas through 0.1M NaOH solution; respectively communicate with 13 CO 2 5min, 10min, 15min and 20 min; the control module controls the first electromagnetic valve to be closed after being opened, simultaneously opens the second electromagnetic valve, closes the lamp source or uses shading cloth to cover the assimilation box, adjusts the flow rate of the flowmeter to 100L/min, ventilates for 2min, 13 CO 2 the marking process ends.
The further technical proposal is that 13 The method for measuring the abundance of C comprises the following steps:
13 CO 2 after marking, the plant is taken out rapidly, liquid nitrogen is used for stopping the tissue organ to be detected, the plant is dried to constant weight by using an oven at the temperature of 80 ℃, a ball mill is used for grinding the tissue sample to be powder state, and an isotope mass spectrometer is used for measuring 13 C abundance, and tissue organ within 5min calculated by using unlabeled sample as control 13 Accumulation amount of C.
Adopt the produced beneficial effect of above-mentioned technical scheme to lie in: 1) the measurable tissues and organs are wide: the traditional photosynthetic gas exchange measuring system can only meet the measurement of the leaf level, not only can the photosynthetic efficiency of different tissues and organs be measured, but also can be used for evaluating the photosynthetic rate of different parts of each tissue and organ.
2) The determination efficiency is high: taking 4-5 leaf stage rice as an example, the length, width and height of the assimilation box are respectively 70cm, 50cm and 30cm for measurement, and 100 rice plants in the assimilation box can be obtained within 5min 13 CO 2 And (4) marking.
3) The marking time is consistent: the traditional photosynthetic gas exchange determination system is limited by the number of analyzers, and the photosynthetic determination time of different plants may differ by hours or days; but the technology can meet the synchronous measurement of the photosynthetic rate of the whole plant and each organ, and further eliminate the system error caused by different measurement time.
Drawings
The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.
FIG. 1 is a functional block diagram of a system according to an embodiment of the present invention;
FIG. 2 is a flow chart of a method according to an embodiment of the present invention;
FIG. 3 illustrates a method according to an embodiment of the present invention 13 A flow chart for C abundance determination;
FIG. 4 is a plot of the marker time in an embodiment of the present invention;
FIG. 5 is a graph showing a leaf sheath after shading treatment according to an embodiment of the present invention;
wherein: 1. a control module; 2. a relay; 3. 13 CO 2 a source of a mixed gas; 4. a first solenoid valve; 5. a second solenoid valve; 6. an air pump; 7. a flow meter; 8. an air agitation device; 9. an environmental parameter measurement sensor; 10. an outdoor daylight source; 11. a gas processing device; 12. a data acquisition line; 13. assimilation box.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.
As shown in fig. 1, the embodiment of the invention discloses a system for synchronously measuring photosynthetic rates of different tissues and organs of a plant, which comprises a control module 1, wherein the control module 1 can use an Arduino or microcontroller such as a single chip microcomputer;
the control output end of the control module 1 is connected with the control input end of the relay 2, and the two relays 2The control output end is respectively connected with the control input ends of the first electromagnetic valve 4 and the second electromagnetic valve 5, and the gas input end of the first electromagnetic valve 4 is connected with the gas input end of the second electromagnetic valve 5 13 CO 2 The mixed gas source 3 is connected, the gas output end of the first electromagnetic valve 4 is connected with one input end of a flow meter 7, the gas input end of the second electromagnetic valve 5 is connected with an air pump 6, the gas output end of the second electromagnetic valve 5 is connected with the other input end of the flow meter 7, the output end of the flow meter is communicated with an assimilation box 13 through a plurality of pipelines, a plant to be tested is arranged in the assimilation box 13, an air stirring device 8 and an environmental parameter measuring sensor 9 are arranged in the assimilation box 13, and preferably, a plurality of fans can be used for the air stirring device 8; the control input end of the air stirring device 8 is connected with the control output end of the control module 1 and is used for acting under the control of the control module 1; the environment parameter measuring sensor 9 is connected with the signal input end of the control module 1 and is used for transmitting measured data to the control module 1 for processing; the artificial light source is positioned at the outer side of the assimilation box, is controlled by the control module 1 and is used for providing illumination for the system; the gas treatment device 11 is connected to an exhaust port of the assimilation box 13 through a pipeline, and the gas discharged from the assimilation box 13 is treated by the gas treatment device and then discharged into the air. Further, the gas treatment device 11 may include a container and a NaOH solution in the container, and the exhausted gas is absorbed by the NaOH solution.
Further, the main principle of the system of the present application is to supply the plants with the above-mentioned ingredients 13 CO 2 The plants will be cultivated by photosynthesis 13 C was assimilated and fixed in plant tissue organs and detected by using an isotope mass spectrometer (Elementar, ISOprime100, Germany) 13 C abundance, and calculating by using plant tissue and organ not labeled by the above mixed gas as control 13 The accumulation amount of C is used for characterizing the photosynthesis rate of each organ tissue of the plant.
Further, the air pump is used for supplying air to the plants in the process of adapting to the environment of the assimilation box and photoinduction of the plants, and 13 CO 2 for assimilating residues in boxes after marking 13 CO 2 Recovering; the gas flow meter is used for controlling and detecting the flow rate of air flowing into the assimilation box.
Further, the Arduino and the relay control the connection state and the connection time of the two electromagnetic valves together; the two electromagnetic valves are respectively connected with 13 CO 2 The air pump and the air mixture of (1).
Further, the assimilation box comprises a lower assimilation chamber and an upper sealing cover; the lower assimilation chamber comprises an air inlet, an air outlet, a fan and a temperature and humidity sensor; the fan is used for mixing the gas in the assimilation box; the environmental factor sensor is arranged on the inner wall of the assimilation box and used for detecting the humidity, the humidity and the change of the photosynthetically active radiation in the assimilation box.
Furthermore, the signal acquisition device is mainly used for acquiring temperature, humidity and photosynthetic active radiation data detected by the environmental factor sensor in the assimilation box.
As shown in FIG. 2, the embodiment of the invention also discloses a method for synchronously measuring the photosynthetic rate of different tissues and organs of a plant, wherein the measuring method uses the measuring system, and the method comprises the following steps:
the method comprises the following steps: test material selection and planting
The method disclosed by the application is used for constructing experiments and verifying that the selected material is the conventional rice variety Huanghuazhan which is widely planted in China. The Huanghuazhan is planted by a plug cultivation mode, the specification of each plug is 5 multiplied by 11cm (length multiplied by width multiplied by height), and a culture medium is commercial nutrient soil. The material is grown in an environment-controllable CONVIRON step-in plant growth box, the temperature of the incubator is controlled to be 28/22 ℃ (day/night), and the illumination intensity of the leaf level is set to be 1000 mu mol m -2 s -1 . Fine management is carried out during planting, and the occurrence of diseases, pests and weeds is strictly controlled. After 17 days of sowing, the rice seedlings are subjected to follow-up 13 CO 2 And (5) labeling experiments.
Step two: construction and Assembly of marking devices
The measurement system is assembled as shown in fig. 1, wherein a control module 1(Arduino) is used for opening and closing and opening and closing control of the first solenoid valve 4 and the second solenoid valve 5, and signal data of an environmental parameter measuring sensor 9 (temperature and humidity sensor) is recorded through a data acquisition line 12. After the plants are placed in the assimilation box 13, an artificial light source or an outdoor sunlight source 10 is started, the Arduino is used for controlling the second electromagnetic valve 5 to be opened, the first electromagnetic valve 4 is closed, and meanwhile, under the control of the flowmeter 7, the air pump 6 enables outside air to enter the assimilation box at the speed of 20L/min to supply gas requirements for normal photosynthesis and respiration of the plants in the assimilation box; meanwhile, the heterogeneity of the concentration and the air temperature of the gas components in the assimilation box is avoided, a plurality of micro fans are used for stirring the air in the assimilation box, and the gas exhausted by the assimilation box is directly exhausted to the atmosphere without passing through the device 11.
Step three: 13 CO 2 determination of a mark time
Photosynthetic gas exchange rates were accomplished using a LI-COR 6800 portable instrument (LI-COR inc., Lincoln, NE, USA). Setting a reference chamber CO of a photosynthetic apparatus 2 The concentration is 400 mu mol -1 VPD is 1.2KPa, and the horizontal illumination intensity of leaves is 1500 mu mol m -2 s -1 . And recording photosynthetic gas exchange data after the variation amplitude of the net photosynthetic rate within 30s is lower than 5%. Subsequently switching the air inlet to the above 13 CO 2 Mixing gas, starting an automatic illumination control program of the photosynthetic apparatus, respectively marking different leaves for 20s, 40s, 60s, 120s, 180s, 240s, 300s, 600s, 1200s and 1800s, and automatically switching the illumination intensity to 0 mu mol m after the respective marking time is finished -2 s -1 . Then cutting leaves in a leaf chamber, photographing for calculating the area of the leaves, quickly placing the leaves in liquid nitrogen, drying the leaves at 80 ℃ to constant weight, and grinding a tissue sample to be in a powder state by using a ball mill for detection.
Step four: 13 CO 2 labelling experiments
Placing the test material prepared in the first step into an assimilation box under the treatment of shading and non-shading of the leaf sheath respectively, and basically adapting the plant body to the environment in the leaf chamber after air is supplied for 30min by an air pump. The second electromagnetic valve 5 is controlled to be closed through Arduino, the first electromagnetic valve 4 is opened, and the flowmeter7 control contains 13 CO 2 The mixed gas enters the assimilation box at the speed of 20L/min, and the gas discharged from the assimilation box absorbs waste gas through 0.1M NaOH solution. Respectively communicate with 13 CO 2 5min, 10min, 15min and 20 min. The Arduino controls the first electromagnetic valve to be closed after the first electromagnetic valve is opened, simultaneously opens the second electromagnetic valve, closes the lamp source or covers the assimilation box by using shading cloth, adjusts the flow rate of the flowmeter to 100L/min, ventilates for 2min, 13 CO 2 the marking process ends.
Step five: 13 c abundance determination
As shown in figure 3 of the drawings, 13 CO 2 after marking, the plant is taken out rapidly, liquid nitrogen is used for stopping the tissue organ to be detected, the plant is dried to constant weight by using an oven at the temperature of 80 ℃, a ball mill is used for grinding the tissue sample to be powder state, and an isotope mass spectrometer is used for measuring 13 C abundance, and tissue organ within 5min calculated by using unlabeled sample as control 13 Accumulation amount of C.
Step six: data analysis
As can be seen from FIG. 4, the marking time is within 20min 13 CO 2 Measured by marking techniques 13 The coincidence degree of the C accumulation rate and the photosynthetic rate measured by the photosynthetic measurement system is high, and after the marking time exceeds 30min, the method and the measured value of the photosynthetic measurement system have large deviation, so that the technology is used for evaluating the photosynthetic rate of the rice 13 CO 2 The labeling time should be less than 20min, and in view of the economy and efficiency of the assay, a labeling time of 5min is recommended.
When the measurement of each tissue and organ is performed, the tissue and organ are verified 13 And C, if the interference is caused, shading and non-shading treatment are respectively carried out on the leaf sheaths. As can be seen from FIG. 5, the light shielding treatment of the leaf sheath is performed within 20min 13 CO 2 The marking does not affect the inside of the leaf sheath 13 And (4) C content. In addition, of leaf sheaths 13 The C accumulation amount has a better linear relation with the marking time. The recommended labeling time is 5min, considering the economy and efficiency of the assay.
The photosynthetic determination system for each tissue organ of a rice single plant is established, the problems that the photosynthetic determination device is low in determination efficiency, non-leaf organ is limited, determination time is inconsistent and the like are solved, the technical blank between the level of photosynthetic determination from a single leaf to each tissue organ is made up, the research on photosynthetic physiological ecology is facilitated, and therefore a high-photosynthetic-efficiency breeding process is promoted.
Claims (10)
1. A system for synchronously measuring photosynthetic rates of different tissues and organs of a plant is characterized by comprising: the control module (1), the control output of control module (1) is connected with the control input of relay (2), two control outputs of relay (2) are connected with the control input of first solenoid valve (4) and second solenoid valve (5) respectively, the gas input of first solenoid valve (4) and 13 CO 2 the device comprises a mixed gas source (3), a gas output end of a first electromagnetic valve (4) is connected with one input end of a flow meter (7), a gas input end of a second electromagnetic valve (5) is connected with an air pump (6), a gas output end of the second electromagnetic valve (5) is connected with the other input end of the flow meter (7), an output end of the flow meter is communicated with an assimilation box (13) through a plurality of pipelines, a plant to be tested is arranged in the assimilation box (13), an air stirring device (8) and an environmental parameter measuring sensor (9) are arranged in the assimilation box (13), and a control input end of the air stirring device (8) is connected with a control output end of a control module (1) and is used for acting under the control of the control module (1); the environment parameter measuring sensor (9) is connected with the signal input end of the control module (1) and is used for transmitting measured data to the control module (1) for processing; the artificial light source is positioned at the outer side of the assimilation box and is controlled by the control module (1) to provide illumination for the system; the gas treatment device (11) is connected with an exhaust port of the assimilation box (13) through a pipeline, and gas exhausted from the assimilation box (13) is treated by the gas treatment device and then exhausted into the air.
2. The system for synchronously measuring photosynthetic rate of different tissues and organs of a plant as claimed in claim 1, wherein: the control module (1) uses Arduino or a single chip microcomputer.
3. The system for synchronously measuring photosynthetic rate of different tissues and organs of a plant as claimed in claim 1, wherein: the air stirring device (8) comprises a plurality of fans.
4. The system for synchronously measuring photosynthetic rate of different tissues and organs of a plant as claimed in claim 1, wherein: the environment parameter measuring sensor (9) is a temperature and humidity sensor.
5. The system for synchronously measuring photosynthetic rate of different tissues and organs of a plant as claimed in claim 1, wherein: the gas treatment device (11) comprises a container and NaOH solution positioned in the container, and the exhausted gas is absorbed by the NaOH solution.
6. A method for synchronously measuring photosynthetic rate of different tissues and organs of a plant, wherein the measuring method uses the measuring system as claimed in any one of claims 1 to 5, and the method comprises the following steps:
selecting and planting test materials;
constructing and assembling a measuring system;
13 CO 2 determining the marking time;
13 CO 2 labelling experiments
13 C abundance determination;
and (6) analyzing the data.
7. The method for synchronously measuring photosynthetic rates of different tissues and organs of a plant as claimed in claim 6, wherein the method comprises 13 CO 2 The method for determining the marking time comprises the following steps:
photosynthetic gas exchange rate was accomplished using LI-COR 6800, setting photosynthetic apparatus reference cell CO 2 The concentration is 400 mu mol -1 VPD of 1.2KPa, leaf levelThe illumination intensity is 1500 mu mol m -2 s -1 (ii) a Recording photosynthetic gas exchange data after the variation amplitude of the net photosynthetic rate within 30s is lower than 5%; then the first solenoid valve (4) and the second solenoid valve (5) are controlled to switch the air inlet to 13 CO 2 Mixing gas, starting an automatic illumination control program of the photosynthetic instrument, marking different leaves for 20s, 40s, 60s, 120s, 180s, 240s, 300s, 600s, 1200s and 1800s respectively, and automatically switching the illumination intensity to 0 mu mol m after marking time is finished respectively - 2 s -1 And then cutting leaves in the assimilation box, photographing for calculating the area of the leaves, quickly placing the leaves in liquid nitrogen, drying at 80 ℃ to constant weight, and grinding a tissue sample to be detected after a powder state by using a ball mill.
8. The method for synchronously measuring photosynthetic rates of different tissues and organs of a plant as claimed in claim 6, wherein the method comprises 13 CO 2 The method of labeling experiments comprises the following steps:
placing prepared test materials in an assimilation box under the treatment of shading and non-shading of leaf sheaths respectively, supplying air for 30min by an air pump (6), allowing the plant body to adapt to the environment in the assimilation box basically, controlling the second electromagnetic valve (5) to close by a control module, opening the first electromagnetic valve (4), and controlling the content of the plant material by a flowmeter (7) 13 CO 2 The mixed gas enters an assimilation box at the speed of 20L/min, and the gas discharged from the assimilation box (13) absorbs waste gas through 0.1M NaOH solution; respectively communicate with 13 CO 2 5min, 10min, 15min and 20 min; the control module (1) controls the first electromagnetic valve (4) to be closed after being opened, simultaneously opens the second electromagnetic valve (5), closes the lamp source or uses shading cloth to cover the assimilation box (13), the flow rate of the flowmeter (7) is adjusted to 100L/min, after 2min of ventilation, 13 CO 2 the marking process ends.
9. The method for synchronously measuring photosynthetic rates of different tissues and organs of a plant as claimed in claim 6, wherein the method comprises 13 The method for measuring the abundance of C comprises the following steps:
13 CO 2 after marking, the plant is taken out rapidly, liquid nitrogen is used for stopping the tissue organ to be detected, the plant is dried to constant weight by using an oven at the temperature of 80 ℃, a ball mill is used for grinding the tissue sample to be powder state, and an isotope mass spectrometer is used for measuring 13 C abundance, and tissue organ within 5min calculated by using unlabeled sample as control 13 Accumulation amount of C.
10. The method for synchronously measuring photosynthetic rates of different tissues and organs of a plant as claimed in claim 7, wherein the labeling time used in the test is 5 min.
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| DE102007031680A1 (en) * | 2007-07-06 | 2009-01-08 | Thermo Fisher Scientific (Bremen) Gmbh | Apparatus for providing gases, in particular for the isotope ratio analysis |
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