CN102453673B - Plant stomata bioanalysis tank and analytical method - Google Patents
Plant stomata bioanalysis tank and analytical method Download PDFInfo
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- CN102453673B CN102453673B CN2010106123238A CN201010612323A CN102453673B CN 102453673 B CN102453673 B CN 102453673B CN 2010106123238 A CN2010106123238 A CN 2010106123238A CN 201010612323 A CN201010612323 A CN 201010612323A CN 102453673 B CN102453673 B CN 102453673B
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- supply system
- cell state
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Abstract
The invention relates to a bioanalysis tank used for researching a response mechanism of plant stomata to environmental stress. The bioanalysis tank comprises a tank body, an air supply system and a cell state detection system and is used for carrying out real-time interference-free detection on the cell state. The bioanalysis tank provided by the invention is dedicated to research on a plant stomata signal transducer mechanism. By applying the bioanalysis tank provided by the invention, solution environment in which plant epidermis is located can be stabilized, test treatment is implemented simply, conveniently and accurately, cell detection and data image storage are synchronously completed, operation simplification, accurate processing and intelligent data collection and sorting are realized, and higher academic application value and commercial value are obtained.
Description
Technical field
The present invention relates to a kind of bioanalysis device, relate in particular to a kind of bioanalysis case of plant stomata to the response mechanism of environment-stress of studying.
Background technology
The pore of plant mainly is distributed in the surface of blade, by 2 guard cells, around forming, is that plant and its ambient air are carried out CO
2with the passage of water vapor exchange, also to plant materials, keep water balance to play an important role.The guard cell can adjust stomatal aperture by integrating inside and outside environmental factor information, thereby keeps water balance and the normal photosynthesis rate of plant.Be tested and appraised the involved in plant pore and reply the gene that environmental factor is relevant, can be the improvement of farm crop anti-adversity and provide fundamental basis.
To the plant stomata detection of dynamic, be mainly to rely on indirect method at present, for example, the leaf temperature that adopts infrared thermoviewer to detect under the varying environment condition changes, indirectly show the state of pore, adopt photosynthetic instrument to monitor the local blade transpiration rate and carry out indirect calculation pore opening degree, or monitor the transpiration rate of whole strain plant with large climatic chamber, infer the dynamic change of pore.
Therefore, lack at present the biochemical state of accurate simulating plant body physiological, conveniently apply environmental factor and process, from the equipment of cell levels monitoring guard cell dynamic change.Given this, Te Famingben equipment.
Summary of the invention
The object of the invention is to set up an accurate controlled plant stomata analytical system, the aperture of studying pore under the environmental factor conditions such as ion, plant hormone and light, CO2 changes, identify that participating in pore replys the signal component of the various cell internal and external environment factors and resolve corresponding molecular regulation network, for the degeneration-resistant molecular improvement of farm crop provides theoretical and genetic resources basis.And provide the analytical procedure of using equipment of the present invention to carry out the plant stomata bioanalysis.
Technical scheme of the present invention is: the present invention proposes a kind of plant stomata bioanalysis case, comprising:
-casing, put into physiological buffer and environmental factor to be studied;
-air supply system, by the air Injection of heterogeneity in physiological buffer;
-cell state detection system, for realizing the real-time noiseless detection to cell state.
Wherein, inject distilled water in casing; Built-in circulating water pump, can stir evenly water; On put sample splint, the embedded circular hole of sample splint, put culture dish in hole; Culture dish covers punching; Pour physiological buffer and environmental factor to be studied in culture dish into.
Concrete, can put the culture dish of 35 diameter 5cm on sample splint.
Preferably, the air supply system that air supply system is the carbonic acid gas filtering or high concentration carbon dioxide supply system.
Wherein, the air supply system of carbonic acid gas filtering mainly consists of pneumatic pump, carbonic acid gas filtering bottle, output gas flow of gas control valve and gas outlet terminal.
The high concentration carbon dioxide supply system comprises carbon dioxide canister, output gas flow of gas controlling valve and gas outlet terminal.
In cell state detection system described in the analysis case, mainly by opticmicroscope, computer, optical gate, camera and each parts joint control software, formed; By the joint control software in computer, optical gate, camera and three kinds of equipment of microscope are set and carry out linkage work, realize the real-time noiseless detection to cell state.
In the phytotron that analysis case of the present invention is controlled at temperature, humidity and light source or greenhouse, use.
Plant stomata bioanalytical method of the present invention, comprise the steps:
(1) the bioanalysis case is injected to distilled water, casing is placed on temperature, humidity and light source controlled phytotron or the culturing rack in greenhouse, guarantees the stable of the interior water temperature of light source and casing.
(2) by the punching of culture dish lid, be placed on the sample splint in casing.
(3) according to the environmental factor difference of research, add different physiological buffers in culture dish.
(4) the gas outlet terminal of air supply system is inserted in the physiological buffer in culture dish, the air of filtering carbonic acid gas (or high concentration carbon dioxide) is pumped in physiological buffer, pump into the flow of gas by gas flow control valve adjustment.
(5) move in advance the bioanalysis case 1 hour, make the indices of physiological buffer keep stable.
(6) plant epidermis that will study is stripped down, put into solution, process 2.5 hours.
(7) utilize the cell state detection system to observe the aperture of plant stomata change, the acknowledgement mechanism of research plant stomata guard cell to certain ambient signal factor in solution.
The present invention integrates environmental simulation, process and to apply and cell detection, is all highly controlled bio-analysis system of process of the test and test conditions.The main application of this equipment comprises:
(1) Plant Guard Cells is to CO
2response studies.
(2) Plant Guard Cells is to Cl
-and Ca
2+isoionic response studies.
(3) response studies of Plant Guard Cells to plant growth hormoness such as ABA, IAA.
(4) response studies of Plant Guard Cells to the environment pH value.
Beneficial effect of the present invention is: the present invention is exclusively used in the research of plant stomata signal transducers mechanism, by the present invention, can make plant epidermis solution environmental of living in stable, the test processing applies easy to be accurate, cell detection and data image storage synchronously complete, realized simple to operateization, process precision, data collection arrangement intellectuality, there is higher academic using value and commercial value.
The accompanying drawing explanation
Fig. 1 is that the present invention uses schematic diagram
Wherein: 1, casing 2, water circulating pump
3, supporting plate 4, circular hole
5, culture dish 6, output gas flow of gas control valve
7, pneumatic pump 8, carbonic acid gas filtering bottle
9, gas outlet terminal 10, carbon dioxide canister
11, opticmicroscope 12, camera
13, the computer 14 of joint control software, electronic optical gate are housed
Fig. 2 is that Abscisic Acid (ABA) suppresses the stomatal opening lab diagram.
Embodiment
Below by embodiment, the present invention will be further described.
Embodiment mono-: the response studies of Plant Guard Cells to plant growth hormoness such as ABA, IAA.
As shown in Figure 1, plant stomata bioanalysis case, comprise casing 1, air supply system, cell state detection system, for realizing the real-time noiseless detection to cell state.Wherein, built-in circulating water pump 2 in casing 1; On put sample splint 3, the embedded circular hole 4 of sample splint 3, put culture dish 5 in hole; Culture dish 5 covers punching; Pour physiological buffer and environmental factor to be studied in culture dish 5 into.35 circular holes 4 are arranged on supporting plate 3, can put the culture dish 5 of 35 diameter 5cm.
Air supply system adopts the air supply system of carbonic acid gas filtering, mainly pneumatic pump 7, carbonic acid gas filtering bottle 8, output gas flow of gas control valve 6 and gas outlet terminal 9, consists of.
The cell state detection system mainly consists of opticmicroscope 11, computer 13, optical gate, camera 12 and each parts joint control software; By the joint control software in computer 13, optical gate 14, camera 12 and 11 3 kinds of equipment of microscope are set and carry out linkage work, realize the real-time noiseless detection to cell state.
(1) the bioanalysis case is injected to distilled water, casing 1 is placed on temperature, humidity and light source controlled phytotron or the culturing rack in greenhouse, guarantees the stable of light source and casing 1 interior water temperature.
(2) by culture dish 5 lid punchings, be placed on the sample splint 3 in casing.
(3) mother liquor of usining containing different concns ABA plant growth hormones is as physiological buffer, add culture dish 5, ABA plant growth hormones mother liquor is that ethanolic soln is solvent, therefore in the present embodiment, also will be with ethanolic soln in contrast, and concentration can not be higher than 0.1%, in case ethanol is as the signaling molecule interference test.
(4) the gas outlet terminal of air supply system is inserted in the physiological buffer in culture dish, the air of filtering carbonic acid gas is pumped in physiological buffer, pump into the flow of gas by gas flow control valve adjustment.
(5) move in advance the bioanalysis case 1 hour, make the indices of physiological buffer keep stable.
(6) plant epidermis that will study is stripped down, put into solution, process 2.5 hours.
(7) utilize the cell state detection system (with the opticmicroscope 11 of optical gate, camera 12, the computer 13 of joint control software is housed) observe the aperture of plant stomata change, the acknowledgement mechanism of research plant stomata guard cell to ABA plant growth hormones in solution.
As shown in Figure 2, wherein X-coordinate is ABA concentration to analytical results, the first transaction of a day's business length that ordinate zou is plant stomata, and the ethanol that contrast is 0.1%, the post of grey is wild-type, the black post is mutant.
Embodiment bis-: Plant Guard Cells is to CO
2response studies.
CO in this experiment
2the detection of concentration and control extremely important.CO in contrast
2-free air and from CO
2the composition of the gas of gas cylinder and temperature all will meet the requirement of experiment.
Plant stomata bioanalysis case, comprise casing 1, air supply system, cell state detection system, for realizing the real-time noiseless detection to cell state.Wherein, built-in circulating water pump 2 in casing 1; On put sample splint 3, the embedded circular hole 4 of sample splint 3, put culture dish 5 in hole; Culture dish 5 covers punching; Pour physiological buffer and environmental factor to be studied in culture dish 5 into.
35 circular holes 4 are arranged on supporting plate 3, can put the culture dish 5 of 35 diameter 5cm.
Air supply system adopts the air supply system of carbonic acid gas filtering, mainly carbon dioxide canister 10, output gas flow of gas control valve 6 and gas outlet terminal 9, consists of.
The cell state detection system mainly forms by being inverted opticmicroscope 11, computer 13, optical gate 14, camera 12 and each parts joint control software; By the joint control software in computer 13, optical gate, camera 12 and 11 3 kinds of equipment of microscope are set and carry out linkage work, realize the real-time noiseless detection to cell state.
(1) the bioanalysis case is injected to distilled water, casing 1 is placed on temperature, humidity and light source controlled phytotron or the culturing rack in greenhouse, guarantees the stable of light source and casing 1 interior water temperature.
(2) by culture dish 5 lid punchings, be placed on the sample splint in casing.
(3) put into physiological buffer in culture dish.
(4) the gas outlet terminal of air supply system is inserted in the physiological buffer in culture dish, high concentration carbon dioxide is pumped in physiological buffer, pump into the flow of gas by gas flow control valve adjustment.
(5) move in advance the bioanalysis case 1 hour, make the indices of physiological buffer keep stable.
(6) plant epidermis that will study is stripped down, put into solution, process 2.5 hours.
(7) utilize the cell state detection system to observe the aperture of plant stomata change, research plant stomata guard cell is to CO
2acknowledgement mechanism.
Embodiment tri-: Plant Guard Cells is to Cl
-and Ca
2+isoionic response studies.
Plant stomata bioanalysis case, comprise casing 1, air supply system, cell state detection system, for realizing the real-time noiseless detection to cell state.Wherein, built-in circulating water pump 2 in casing 1; On put sample splint 3, the embedded circular hole 4 of sample splint 3, put culture dish 5 in hole; Culture dish 5 covers punching; Pour physiological buffer and environmental factor to be studied in culture dish 5 into.
35 circular holes 4 are arranged on supporting plate 3, can put the culture dish 5 of 35 diameter 5cm.
Air supply system adopts the air supply system of carbonic acid gas filtering, mainly pneumatic pump 7, carbonic acid gas filtering bottle 8, output gas flow of gas control valve 6 and gas outlet terminal 9, consists of.
The cell state detection system mainly forms by being inverted opticmicroscope 11, computer 13, optical gate, camera 12 and each parts joint control software; By the joint control software in computer 13, optical gate 14, camera 12 and 11 3 kinds of equipment of microscope are set and carry out linkage work, realize the real-time noiseless detection to cell state.
(1) the bioanalysis case is injected to distilled water, casing 1 is placed on temperature, humidity and light source controlled phytotron or the culturing rack in greenhouse, guarantees the stable of light source and casing 1 interior water temperature.
(2) by culture dish 5 lid punchings, be placed on the sample splint in casing.
(3) to contain Ca
2+isoionic solution, as physiological buffer, adds culture dish.Ionic concn can not be too high, avoids the murder by poisoning reaction to cell.
(4) the gas outlet terminal of air supply system is inserted in the physiological buffer in culture dish, the air of filtering carbonic acid gas is pumped in physiological buffer, pump into the flow of gas by gas flow control valve adjustment.
(5) move in advance bioanalysis case at least 30 minutes, make the indices of physiological buffer keep stable.
(6) plant epidermis that will study is stripped down, put into solution, process 2.5 hours.
(7) utilize the cell state detection system to observe the aperture of plant stomata change, research plant stomata guard cell is to Cl
-and Ca
2+isoionic acknowledgement mechanism.
Embodiment 4: the response studies of Plant Guard Cells to the environment pH value.
Usually understand the variation with pH value during the guard cell replys ambient signal, but the effect of PH in this reaction also known little about it at present.Native system is very suitable for carrying out correlative study.
Plant stomata bioanalysis case, comprise casing 1, air supply system, cell state detection system, for realizing the real-time noiseless detection to cell state.Wherein, built-in circulating water pump 2 in casing 1; On put sample splint 3, the embedded circular hole 4 of sample splint 3, put culture dish 5 in hole; Culture dish 5 covers punching; Pour physiological buffer and environmental factor to be studied in culture dish 5 into.
35 circular holes 4 are arranged on supporting plate 3, can put the culture dish 5 of 35 diameter 5cm.
Air supply system adopts the air supply system of carbonic acid gas filtering, mainly pneumatic pump 7, carbonic acid gas filtering bottle 8, output gas flow of gas control valve 6 and gas outlet terminal 9, consists of.
The cell state detection system mainly forms by being inverted opticmicroscope 11, computer 13, optical gate, camera 12 and each parts joint control software; By the joint control software in computer 13, optical gate 14, camera 12 and 11 3 kinds of equipment of microscope are set and carry out linkage work, realize the real-time noiseless detection to cell state.
(1) the bioanalysis case is injected to distilled water, casing 1 is placed on temperature, humidity and light source controlled phytotron or the culturing rack in greenhouse, guarantees the stable of light source and casing 1 interior water temperature.
(2) by culture dish 5 lid punchings, be placed on the sample splint in casing.
(3) using containing the solution with the compound of pH value as physiological buffer, add culture dish.Note to regulate other ion in the compound of pH value (Cl for example
-and Na
+interference effect).
(4) the gas outlet terminal of air supply system is inserted in the physiological buffer in culture dish, the air of filtering carbonic acid gas is pumped in physiological buffer, pump into the flow of gas by gas flow control valve adjustment.
(5) move in advance bioanalysis case one hour, make the indices of physiological buffer keep stable.
(6) plant epidermis that will study is stripped down, put into solution, process 2.5 hours.
(7) utilize the cell state detection system to observe the aperture of plant stomata change, the acknowledgement mechanism of research plant stomata guard cell to the environment pH value.
Claims (6)
1. a plant stomata bioanalysis case, is characterized in that, comprising:
-casing, inside establish water circulating pump and sample splint, and the culture dish of placing in supporting plate can be poured physiological buffer and environmental factor to be studied into;
-air supply system, by the air Injection of heterogeneity in physiological buffer;
-cell state detection system, for realizing the real-time noiseless detection to cell state;
-inject distilled water in casing; Start water circulating pump, water can be stirred evenly; On put sample splint, the embedded circular hole of sample splint, put culture dish in hole, can put the culture dish of 35 diameter 5cm on described sample splint; Culture dish covers punching; Pour physiological buffer and environmental factor to be studied in culture dish into.
2. a kind of plant stomata bioanalysis case according to claim 1, is characterized in that: the air supply system that described air supply system is the carbonic acid gas filtering or high concentration carbon dioxide supply system.
3. a kind of plant stomata bioanalysis case according to claim 2, it is characterized in that: the air supply system of described carbonic acid gas filtering mainly consists of pneumatic pump, carbonic acid gas filtering bottle, output gas flow of gas control valve and gas outlet terminal.
4. a kind of plant stomata bioanalysis case according to claim 2, it is characterized in that: described high concentration carbon dioxide supply system comprises carbon dioxide canister, output gas flow of gas controlling valve and gas outlet terminal.
5. a kind of plant stomata bioanalysis case according to claim 1, it is characterized in that: described cell state detection system mainly consists of opticmicroscope, computer, optical gate, camera and each parts joint control software; By the joint control software in computer, optical gate, camera and three kinds of equipment of microscope are set and carry out linkage work, realize the real-time noiseless detection to cell state.
6. a kind of plant stomata bioanalysis case according to claim 1, is characterized in that: in the phytotron that described analysis case is controlled at temperature, humidity and light source or greenhouse, use.
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| CN108398429B (en) * | 2018-03-07 | 2020-08-04 | 天津农学院 | Plant stomata monitoring system and method for analyzing response of plant stomata to environment by using same |
| CN108469434B (en) * | 2018-04-11 | 2022-01-25 | 山东农业大学 | Device and method for monitoring whether fruit trees lack water |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2204418Y (en) * | 1994-06-03 | 1995-08-02 | 北京农业大学 | Photosynthetic transpiration instrument |
| CN200989897Y (en) * | 2005-06-28 | 2007-12-12 | 北京益康农科技发展有限公司 | Photosynthetic transpiration analyzer |
-
2010
- 2010-12-29 CN CN2010106123238A patent/CN102453673B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2204418Y (en) * | 1994-06-03 | 1995-08-02 | 北京农业大学 | Photosynthetic transpiration instrument |
| CN200989897Y (en) * | 2005-06-28 | 2007-12-12 | 北京益康农科技发展有限公司 | Photosynthetic transpiration analyzer |
Non-Patent Citations (6)
| Title |
|---|
| FGC-2型14C植物光合速率测量装置;冯春生等;《核农学报》;19871231;第1卷(第2期);第105-111页 * |
| 光合作用中光、温、湿和CO2检测装置介绍;方志伟等;《植物生理学通讯》;19901231(第2期);第53-56页 * |
| 冯春生等.FGC-2型14C植物光合速率测量装置.《核农学报》.1987,第1卷(第2期), |
| 国外光合作用测定装置介绍;林植芳;《广西植物》;19841231;第4卷(第3期);第265-268页 * |
| 方志伟等.光合作用中光、温、湿和CO2检测装置介绍.《植物生理学通讯》.1990,(第2期),第53-56页. |
| 林植芳.国外光合作用测定装置介绍.《广西植物》.1984,第4卷(第3期),第265－268页. |
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