CN108307926A - A kind of arbuscular mycorrhizal fungi15N isotope labellings device and production method - Google Patents
A kind of arbuscular mycorrhizal fungi15N isotope labellings device and production method Download PDFInfo
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- CN108307926A CN108307926A CN201810073591.3A CN201810073591A CN108307926A CN 108307926 A CN108307926 A CN 108307926A CN 201810073591 A CN201810073591 A CN 201810073591A CN 108307926 A CN108307926 A CN 108307926A
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- isotope
- growth room
- mycelia
- plant
- mycelia growth
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/0098—Plants or trees
Abstract
The invention discloses a kind of arbuscular mycorrhizal fungi15N isotope labelling device and methods, including mycelia growth room, isotope injection pipe, mycelia growth room opening are covered with filter, and the filter is fixed with filter locking ring.Isotope injection pipe is connect with mycelia growth room by fixed angle with angle correction device, in junction, hot melt adhesive bonds secured, and mycelia growth room is made to be connected with the inclination at 45 ° of isotope injection pipe, meanwhile, keep isotope injection pipe with ground level at vertical relation.Can accurate quantification AMF mycelia to the relative contribution of nitrogen absorbed by plant, it is easily operated, make it is simple, convenient for experiment setting.Meanwhile it is so effective that solve nutritive solution extravasation diffusion problem, it is widely used in indoor and outdoors experiment, eliminates the limitation that external environment carries out experiment, is convenient for the research that plant absorbs nutrient by using AMF mycelia.
Description
Technical field
The present invention relates to a kind of microorganism nitrogen element absorption device more particularly to a kind of arbuscular mycorrhizal fungi15N isotope marks
Remember device and production method.
Background technology
Arbuscular mycorrhizal fungi (Arbuscular Mycorrhizal Fungi, abbreviation AMF) is to be distributed in soil most extensively
One kind and the microorganism of plant obligate symbiosis, in meadow, forest and farmland ecosystem, 80% or more plant all can be with
AMF forms the relationship of mutualistic symbiosis.AMF is one of the microbial functional group that ecological functions are mostly important in soil, to vegetation
Nutrient Cycling, productivity and stability all play important regulating and controlling effect.The mycelia network that AMF is formed can help plant absorption
The soil nutrients such as nitrogen, phosphorus, the water regime for improving plant improve plant drought resistance, enhance plant disease-resistant ability, improve soil knot
Structure etc..
In order to study different plant induced synthesis AMF function difference, quantization AMF mycelia plant nitrogen is absorbed it is opposite
Contribution usually uses locellus separation screen device.Since being put forward for the first time filter locellus system from 1973, the locellus culture of AMF is always
It is the key technology that people probe into, however, traditional locellus separation screen device is in a parallel set, is easy the shadow by other extraneous mushrooms
It rings, mycelia growth room cannot be isolated from the outside completely.Meanwhile locellus device is only adapted to laboratory test progress, to plant itself
Growing environment disturbance is larger, cannot completely separate root system of plant and AMF mycelia growth room, between root system of plant and AMF mycelia
There are still elements diffusion extravasation problems, are unfavorable for the relative contribution that accurate quantification AMF mycelia absorbs plant nitrogen.
Currently, outdoor and indoor universal AMF15N isotope labelling devices fail to be used widely, and pass through profit to plant
The research that nutrient is absorbed with AMF mycelia brings limitation.
Invention content
The object of the present invention is to provide a kind of arbuscular mycorrhizal fungi15N isotope labellings device and production method.
The purpose of the present invention is what is be achieved through the following technical solutions：
The arbuscular mycorrhizal fungi of the present invention15N isotope labelling devices, including mycelia growth room, the mycelia growth room
Side wall is connected with the inclination at 45 ° of isotope injection pipe, and mycelia growth room opening is covered with filter, the filter filter
Locking ring is fixed.
The above-mentioned arbuscular mycorrhizal fungi of the present invention15The production method of N isotope labelling devices, including step：
Step 1：Mycelia growth room is positioned over mycelia growth room fixed area with angle correction device, is grown apart from mycelia
A hole is bored at the opening 3cm of room, diameter 0.3mm is identical as isotope injection nozzle diameter；
Step 2：Isotope injection pipe is connect with mycelia growth room by fixed angle with angle correction device, in junction
It is secured with hot melt adhesive bonding, so that mycelia growth room is connected with the inclination at 45 ° of isotope injection pipe, meanwhile, keep isotope injection
Pipe is with ground level at vertical relation；
Step 3：Indoor pot experiment plant or outdoor wild test plant are chosen, determines plant rhizosphere growth district；
Step 4：After the appropriate soil sieving removal root segment in plant growth region, sterilizes through high pressure high humidity and be packed into mycelia
Growth room；
Step 5：The filter in two kinds of apertures is overlayed on into mycelia growth room bottleneck by experimental design requirement respectively, is sealed with filter
Lock ring fixes filter, and rotates filter locking ring, screw thread and mycelia growth room bottleneck groove tight fits, isotope labelling dress
It sets and completes；
Step 6：Isotope labelling device is kept to tilt 45 ° of inclinations with ground level, meanwhile, isotope injection pipe and ground
Plane is embedded in the depths plant root growth 20cm, in-situ soil is overlying on isotope labelling device at vertical relation, keeps plant
Grow primal environment；
Step 7：Plant will contain 2mg/L after growth after a period of time15The solution of N isotopes is along isotope injection
Pipe is injected into the middle part of mycelia growth room soil, and space-number in the future, collects Plant aboveground and under ground portion, carries out isotopic content
It measures.
As seen from the above technical solution provided by the invention, arbuscular mycorrhizal fungi provided in an embodiment of the present invention15N is same
The plain labelling apparatus in position and production method, can accurate quantification AMF mycelia to the relative contribution of nitrogen absorbed by plant, it is easily operated, make
Simply, it is convenient for experiment setting.
Description of the drawings
Fig. 1 is the structural schematic diagram of isotope injection pipe in the embodiment of the present invention.
Fig. 2 a and Fig. 2 b are respectively the structural schematic diagram of two kinds of filters in the embodiment of the present invention.
Fig. 3 is the structural schematic diagram of filter locking ring in the embodiment of the present invention.
Fig. 4 is arbuscular mycorrhizal fungi provided in an embodiment of the present invention15N isotope labellings device and production method structural representation
Figure.
Fig. 5 is the structural schematic diagram of angle correction device in the embodiment of the present invention.
Fig. 6 is angle correction device vertical view in the embodiment of the present invention.
Fig. 7 is that angle correction device corrects mycelia growth room and isotope labelling tube angulation schematic diagram in the embodiment of the present invention.
Fig. 8 is arbuscular mycorrhizal fungi in inventive embodiments15N isotope labelling devices bury schematic diagram.
Specific implementation mode
The embodiment of the present invention will be described in further detail below.What is be not described in detail in the embodiment of the present invention is interior
Appearance belongs to the prior art well known to professional and technical personnel in the field.
The arbuscular mycorrhizal fungi of the present invention15N isotope labelling devices, preferable specific implementation mode are：
Including mycelia growth room, side wall and the inclination at 45 ° of isotope injection pipe of the mycelia growth room connect, the bacterium
Silk growth room opening is covered with filter, and the filter is fixed with filter locking ring.
Placement status of the device in underground be：
The mycelia growth room opening tilts upward and ground level inclination at 45 °, the isotope injection pipe and ground level
At vertical relation.
The mycelia growth room is 150mlPE vinyon bottles.
The filter locking ring is to remove top and screwed bottle cap, with the plastic bottle opening for making mycelia growth room
Groove phase tight fits.
The isotope injection pipe is 1ml plastics aseptic injection pipes, nozzle diameter 0.3mm.
It is 1mm sieves and aperture is 30 μm of nylon wires that the filter, which includes aperture,.
The above-mentioned arbuscular mycorrhizal fungi of the present invention15The production method of N isotope labelling devices is preferably embodied
Mode is：
Including step：
Step 1：Mycelia growth room is positioned over mycelia growth room fixed area with angle correction device, is grown apart from mycelia
A hole is bored at the opening 3cm of room, diameter 0.3mm is identical as isotope injection nozzle diameter；
Step 2：Isotope injection pipe is connect with mycelia growth room by fixed angle with angle correction device, in junction
It is secured with hot melt adhesive bonding, so that mycelia growth room is connected with the inclination at 45 ° of isotope injection pipe, meanwhile, keep isotope injection
Pipe is with ground level at vertical relation；
Step 3：Indoor pot experiment plant or outdoor wild test plant are chosen, determines plant rhizosphere growth district；
Step 4：After the appropriate soil sieving removal root segment in plant growth region, sterilizes through high pressure high humidity and be packed into mycelia
Growth room；
Step 5：The filter in two kinds of apertures is overlayed on into mycelia growth room bottleneck by experimental design requirement respectively, is sealed with filter
Lock ring fixes filter, and rotates filter locking ring, screw thread and mycelia growth room bottleneck groove tight fits, isotope labelling dress
It sets and completes；
Step 6：Isotope labelling device is kept to tilt 45 ° of inclinations with ground level, meanwhile, isotope injection pipe and ground
Plane is embedded in the depths plant root growth 20cm, in-situ soil is overlying on isotope labelling device at vertical relation, keeps plant
Grow primal environment；
Step 7：Plant will contain 2mg/L after growth after a period of time15The solution of N isotopes is along isotope injection
Pipe is injected into the middle part of mycelia growth room soil, and space-number in the future, collects Plant aboveground and under ground portion, carries out isotopic content
It measures.
The angle correction equipment material is PVC foam plate, takes angle with angulation tolerance and cutting is bonded, including mycelia
Growth room's fixed area and isotope injection pipe clip plate.
The arbuscular mycorrhizal fungi of the present invention15N isotope labellings device and production method, energy accurate quantification AMF mycelia is to planting
Object absorbs the relative contribution of N.
Specific embodiment：
As shown in Figures 1 to 8, including mycelia growth room, isotope injection pipe, filter, filter locking ring, angle correction
Device.The mycelia growth room opening tilts upward at 45 ° with ground level；The mycelia growth room and isotope injection pipe are at 45 °
Angle tilts connection；The filter is overlying on mycelia growth room opening, is fixed with the filter locking ring；The angle correction device
For correcting mycelia growth room and isotope injection pipe clamp angle, meanwhile, keep isotope injection pipe with ground level at vertical relation.
It is further：The mycelia growth room is 150mlPE vinyon bottles.
It is further：The isotope injection pipe is 1ml plastics aseptic injection pipes, nozzle diameter 0.3mm.
It is further：The filter is divided to two kinds, and aperture is respectively 1mm sieves and 30 μm of nylon wires.
It is further：The filter locking ring is to remove top and screwed bottle cap, with making mycelia growth room
Plastic bottle port recess phase tight fits.
It is further：The angle correction equipment material is PVC foam plate, accurately measures angle with protractor, cutting is viscous
It closes, is divided into mycelia growth room fixed area and isotope injection pipe clip plate.
Arbuscular mycorrhizal fungi15N isotope labellings device and production method, step are：Step 1：With angle correction device by bacterium
Silk growth room is fixed on mycelia growth room fixed area, in one hole of brill at mycelia growth room opening 3cm, diameter 0.3mm, with
Isotope injection nozzle diameter is identical；Step 2：Isotope injection pipe and mycelia growth room are pressed into fixed angles with angle correction device
Degree connection, in junction, hot melt adhesive bonds secured, and mycelia growth room is made to be connected with the inclination at 45 ° of isotope injection pipe, meanwhile,
Keep isotope injection pipe with ground level at vertical relation；Step 3：Choose indoor pot experiment plant or outdoor wild examination
Plant is tested, determines plant rhizosphere growth district；Step 4：After the appropriate soil sieving removal root segment in plant growth region, warp
The sterilizing of high pressure high humidity is packed into mycelia growth room；Step 5：The filter in two kinds of apertures is overlayed on into mycelia respectively by experimental design requirement
Filter is fixed with filter locking ring, and rotates filter locking ring by growth room's bottleneck, and screw thread and mycelia growth room bottleneck groove are tight
It closes closely, isotope labelling device completes；Step 6：By the holding of isotope labelling device and ground level inclination at 45 °, bury
In the depths plant root growth 20cm, in-situ soil is overlying on isotope labelling device, keeps plant growth primal environment.Step
Seven：Plant will contain 2mg/L after growth after a period of time15The solution of N isotopes is injected into mycelia along isotope injection pipe
The middle part of growth room's soil, space-number in the future, collect Plant aboveground and under ground portion, carry out isotopic content measurement.
Beneficial effects of the present invention are:
1, isotope labelling device production method is simple, is easy to grasp.
2, isotope labelling device is cheap using material, can it is sterilized after repeatedly use, indoors can be with outdoor
Extensive use.
3, isotope labelling device and ground level tilt 45 ° and filter blocks band and mycelia growth room bottleneck is closely cut
It closes, solves the problems, such as nutritive solution extravasation diffusion, eliminate the test error as caused by solution extravasation diffusion.
4, the fixed filter operation of filter block band is simple, and block is close, can be by mycelia growth room using 30 μm of nylon wires
Effectively separate with plant root growth space.
5, filter uses two kinds of apertures of 1mm sieves and 30 μm of nylon wires, wherein 1mm sieves can be tiny by plant simultaneously
Root system and AMF mycelia, and 30 μm of nylon wires only allow AMF mycelia to pass through, by measuring Plant aboveground and underground isotopic content,
It can show that plant only relies on AMF mycelia absorbed nitrogen amount of element and accounts for plant by tiny root system and AMF mycelia absorbed nitrogen amount of element
Ratio, to quantify relative contribution of the AMF mycelia to nitrogen absorbed by plant.
6, setting isotope labelling device disturbs plant growth environment small, meanwhile, mycelia growth room and external environment
Isolation eliminates other influences of the extraneous mushroom to experiment.
In conclusion the arbuscular mycorrhizal fungi of the present invention15N isotope labellings device and production method, can accurate quantification AMF
Mycelia is to the relative contribution of nitrogen absorbed by plant, and easily operated, making is simple, convenient for experiment setting.Meanwhile it is so effective that solve battalion
Solution extravasation diffusion problem is supported, indoor and outdoors experiment is widely used in, the limitation that external environment carries out experiment is eliminated, is convenient for
Carry out the research that plant absorbs nutrient by using AMF mycelia.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Any one skilled in the art is in the technical scope of present disclosure, the change or replacement that can be readily occurred in,
It should be covered by the protection scope of the present invention.Therefore, protection scope of the present invention should be with the protection model of claims
Subject to enclosing.
Claims (8)
1. a kind of arbuscular mycorrhizal fungi15N isotope labelling devices, which is characterized in that including mycelia growth room, the mycelia growth
The side wall of room is connected with the inclination at 45 ° of isotope injection pipe, and mycelia growth room opening is covered with filter, and the filter is used
Filter locking ring is fixed.
2. arbuscular mycorrhizal fungi according to claim 115N isotope labelling devices, which is characterized in that the device is in underground
Placement status be：
The mycelia growth room opening is tilted upward with ground level inclination at 45 °, the isotope injection pipe and ground level at vertical
Straight relationship.
3. arbuscular mycorrhizal fungi according to claim 215N isotope labelling devices, which is characterized in that the mycelia growth
Room is 150mlPE vinyon bottles.
4. arbuscular mycorrhizal fungi according to claim 315N isotope labelling devices, which is characterized in that the filter block
Ring is to remove top and screwed bottle cap, with the plastic bottle port recess phase tight fits for making mycelia growth room.
5. arbuscular mycorrhizal fungi according to claim 215N isotope labelling devices, which is characterized in that the isotope note
It is 1ml plastics aseptic injection pipes, nozzle diameter 0.3mm to penetrate pipe.
6. arbuscular mycorrhizal fungi according to claim 215N isotope labelling devices, which is characterized in that the filter includes
Aperture is 1mm sieves and aperture is 30 μm of nylon wires.
7. a kind of claim 1 to 6 any one of them arbuscular mycorrhizal fungi15The production method of N isotope labelling devices, it is special
Sign is, including step：
Step 1：Mycelia growth room is positioned over mycelia growth room fixed area with angle correction device, is opened apart from mycelia growth room
A hole is bored at mouth 3cm, diameter 0.3mm is identical as isotope injection nozzle diameter；
Step 2：Isotope injection pipe is connect with mycelia growth room by fixed angle with angle correction device, in junction heat
Melten gel bonding is secured, and mycelia growth room is made to be connected with inclinations at 45 ° of isotope injection pipe, meanwhile, keep isotope injection pipe and
Ground level is at vertical relation；
Step 3：Indoor pot experiment plant or outdoor wild test plant are chosen, determines plant rhizosphere growth district；
Step 4：After the appropriate soil sieving removal root segment in plant growth region, sterilizes through high pressure high humidity and be packed into mycelia growth
Room；
Step 5：The filter in two kinds of apertures is overlayed on into mycelia growth room bottleneck respectively by experimental design requirement, with filter locking ring
Filter is fixed, and rotates filter locking ring, screw thread and mycelia growth room bottleneck groove tight fits, isotope labelling device system
It completes；
Step 6：Isotope labelling device is kept to tilt 45 ° of inclinations with ground level, meanwhile, isotope injection pipe and ground level
At vertical relation, it is embedded in the depths plant root growth 20cm, in-situ soil is overlying on isotope labelling device, keeps plant growth
Primal environment；
Step 7：Plant will contain 2mg/L after growth after a period of time15The solution of N isotopes is noted along isotope injection pipe
It is mapped to the middle part of mycelia growth room soil, space-number in the future, collects Plant aboveground and under ground portion, carries out isotopic content survey
It is fixed.
8. arbuscular mycorrhizal fungi according to claim 715N isotope labellings device and production method, which is characterized in that institute
It is PVC foam plate to state angle correction equipment material, takes angle with angulation tolerance and cutting is bonded, including mycelia growth room is fixed
Area and isotope injection pipe clip plate.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810073591.3A CN108307926B (en) | 2018-01-25 | 2018-01-25 | A kind of arbuscular mycorrhizal fungi15N isotope labelling device and production method |
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| CN201810073591.3A CN108307926B (en) | 2018-01-25 | 2018-01-25 | A kind of arbuscular mycorrhizal fungi15N isotope labelling device and production method |
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| CN108307926A true CN108307926A (en) | 2018-07-24 |
| CN108307926B CN108307926B (en) | 2019-08-09 |
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| CN203999584U (en) * | 2014-08-06 | 2014-12-10 | 兰州大学 | Combined type bush mycorrhizal fungi locellus culture apparatus |
| CN204291868U (en) * | 2014-12-08 | 2015-04-29 | 兰州大学 | Artificial medium is adopted to carry out the locellus culture apparatus of bush mycorrhizal fungi pure culture |
| CN105675705A (en) * | 2016-02-25 | 2016-06-15 | 浙江大学 | Device and method for measuring moving rule of amino acid in soil |
| CN205993154U (en) * | 2016-08-17 | 2017-03-08 | 四川农业大学 | Mycorrhizal fungi locellus culture apparatus |
| CN106834087A (en) * | 2017-03-01 | 2017-06-13 | 厦门大学 | A kind of AM fungies of timing cycle expand numerous and functional experiment device and application method |
| CN107041235A (en) * | 2017-03-27 | 2017-08-15 | 贵州大学 | Utilize nutrient device between atmospheric carbon dioxide regulation and control mycorrhizal fungi transfering plant |
| CN206440716U (en) * | 2017-01-18 | 2017-08-25 | 云南农业大学 | It is a kind of to study the experimental rig that AMF mycelia influences on soil eluviation |
-
2018
- 2018-01-25 CN CN201810073591.3A patent/CN108307926B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN203999584U (en) * | 2014-08-06 | 2014-12-10 | 兰州大学 | Combined type bush mycorrhizal fungi locellus culture apparatus |
| CN204291868U (en) * | 2014-12-08 | 2015-04-29 | 兰州大学 | Artificial medium is adopted to carry out the locellus culture apparatus of bush mycorrhizal fungi pure culture |
| CN105675705A (en) * | 2016-02-25 | 2016-06-15 | 浙江大学 | Device and method for measuring moving rule of amino acid in soil |
| CN205993154U (en) * | 2016-08-17 | 2017-03-08 | 四川农业大学 | Mycorrhizal fungi locellus culture apparatus |
| CN206440716U (en) * | 2017-01-18 | 2017-08-25 | 云南农业大学 | It is a kind of to study the experimental rig that AMF mycelia influences on soil eluviation |
| CN106834087A (en) * | 2017-03-01 | 2017-06-13 | 厦门大学 | A kind of AM fungies of timing cycle expand numerous and functional experiment device and application method |
| CN107041235A (en) * | 2017-03-27 | 2017-08-15 | 贵州大学 | Utilize nutrient device between atmospheric carbon dioxide regulation and control mycorrhizal fungi transfering plant |
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