CN108307926B - A kind of arbuscular mycorrhizal fungi15N isotope labelling device and production method - Google Patents

A kind of arbuscular mycorrhizal fungi15N isotope labelling device and production method Download PDF

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CN108307926B
CN108307926B CN201810073591.3A CN201810073591A CN108307926B CN 108307926 B CN108307926 B CN 108307926B CN 201810073591 A CN201810073591 A CN 201810073591A CN 108307926 B CN108307926 B CN 108307926B
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isotope
growth room
mycelia
injection pipe
plant
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CN108307926A (en
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纪宝明
董强
王平
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Beijing Forestry University
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Beijing Forestry University
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Abstract

The invention discloses a kind of arbuscular mycorrhizal fungi15N isotope labelling device and method, 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, is bonded securely in junction with hot melt adhesive, mycelia growth room and the inclination at 45 ° of isotope injection pipe are connected, meanwhile, keep isotope injection pipe and ground level at vertical relation.Can accurate quantification AMF mycelia to the relative contribution of nitrogen absorbed by plant, easily operated, production is simple, convenient for test setting.Meanwhile it is so effective that solve nutritive solution extravasation diffusion problem, it is widely used in indoor and outdoors and tests, eliminate the limitation that external environment carries out test, be convenient for plant and pass through the research using AMF mycelia absorption nutrient.

Description

A kind of arbuscular mycorrhizal fungi15N isotope labelling device and production method
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 mark Remember device and production method.
Background technique
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 The relationship of AMF formation 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 device fails to be used widely, and passes through benefit to plant Limitation is brought with the research that AMF mycelia absorbs nutrient.
Summary of the invention
The object of the present invention is to provide a kind of arbuscular mycorrhizal fungi15N isotope labelling device and production method.
The purpose of the present invention is what is be achieved through the following technical solutions:
Arbuscular mycorrhizal fungi of the invention15N isotope labelling device, including mycelia growth room, the mycelia growth room Side wall and the inclination at 45 ° of isotope injection pipe connect, and mycelia growth room opening is covered with filter, the filter filter Locking ring is fixed.
Above-mentioned arbuscular mycorrhizal fungi of the invention15The production method of N isotope labelling device, comprising steps of
Step 1: mycelia growth room is placed in 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 Securely with hot melt adhesive bonding, mycelia growth room and the inclination at 45 ° of isotope injection pipe are connected, meanwhile, keep isotope injection Pipe and ground level are at vertical relation；
Step 3: indoor pot experiment plant or outdoor wild test plant are chosen, determines plant rhizosphere growth district；
Step 4: it after the appropriate soil sieving removal root segment in plant growth region, sterilizes through high pressure high humidity and is 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 being 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 isotope 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 Measurement.
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 relative contribution of the accurate quantification AMF mycelia to nitrogen absorbed by plant, easily operated, production Simply, convenient for test setting.
Detailed description of the invention
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 labelling 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 top 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 device buries schematic diagram.
Specific embodiment
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.
Arbuscular mycorrhizal fungi of the invention15N isotope labelling device, preferable specific embodiment is:
Including mycelia growth room, side wall and isotope injection the pipe inclination at 45 ° of the mycelia growth room are connected, 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 is:
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 bottle.
The filter locking ring is to remove top and screwed bottle cap, the plastic bottle opening with production mycelia growth room Groove phase tight fits.
The isotope injection pipe is 1ml plastics aseptic injection pipe, nozzle diameter 0.3mm.
It is 1mm sieve and aperture is 30 μm of nylon wires that the filter, which includes aperture,.
Above-mentioned arbuscular mycorrhizal fungi of the invention15The production method of N isotope labelling device is preferably embodied Mode is:
Comprising steps of
Step 1: mycelia growth room is placed in 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 Securely with hot melt adhesive bonding, mycelia growth room and the inclination at 45 ° of isotope injection pipe are connected, meanwhile, keep isotope injection Pipe and ground level are at vertical relation；
Step 3: indoor pot experiment plant or outdoor wild test plant are chosen, determines plant rhizosphere growth district；
Step 4: it after the appropriate soil sieving removal root segment in plant growth region, sterilizes through high pressure high humidity and is 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 being 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 isotope 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 Measurement.
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.
Arbuscular mycorrhizal fungi of the invention15N isotope labelling device and production method, energy accurate quantification AMF mycelia is to plant The relative contribution of object absorption 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 inclination 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 and ground level at vertical relation.
Further, the mycelia growth room is 150mlPE vinyon bottle.
Further, the isotope injection pipe is 1ml plastics aseptic injection pipe, nozzle diameter 0.3mm.
Further, the filter is divided to two kinds, aperture is respectively 1mm sieve and 30 μm of nylon wires.
Further, the filter locking ring is to remove top and screwed bottle cap, with production mycelia growth room Plastic bottle port recess phase tight fits.
Further, the angle correction equipment material is PVC foam plate, angle is accurately measured 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 labelling device and production method, step are as follows: step 1: use 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 is bonded securely in junction with hot melt adhesive, connects mycelia growth room and the inclination at 45 ° of isotope injection pipe, meanwhile, Keep isotope injection pipe and ground level at vertical relation；Step 3: indoor pot experiment plant or outdoor wild examination are chosen 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 by experimental design requirement respectively 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: the holding of isotope labelling device and ground level inclination at 45 ° are buried 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 isotope 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.
The invention has the benefit that
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 It is widely applied.
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 sieve and 30 μm of nylon wires, wherein 1mm sieve 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 be concluded that plant, which only relies on AMF mycelia absorbed nitrogen amount of element, accounts for plant by tiny root system and AMF mycelia absorbed nitrogen amount of element Ratio, to quantify AMF mycelia to the relative contribution of 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 test.
In conclusion arbuscular mycorrhizal fungi of the invention15N isotope labelling device and production method, can accurate quantification AMF Mycelia is to the relative contribution of nitrogen absorbed by plant, and easily operated, production is simple, convenient for test setting.Meanwhile it is so effective that solve battalion Solution extravasation diffusion problem is supported, indoor and outdoors test is widely used in, the limitation that external environment carries out test is eliminated, is convenient for It carries out plant and passes through the research using AMF mycelia absorption nutrient.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto, Within the technical scope of the present disclosure, any changes or substitutions that can be easily thought of by anyone skilled in the art, 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 (6)

1. a kind of arbuscular mycorrhizal fungi15The production method of N isotope labelling device, it is characterised in that:
The arbuscular mycorrhizal fungi15N isotope labelling device includes mycelia growth room, the side wall of the mycelia growth room and same position Plain injection-tube inclination connection at 45 °, mycelia growth room opening are covered with filter, and the filter is solid with filter locking ring It is fixed；
Placement status of the device in underground is:
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；
The arbuscular mycorrhizal fungi15The production method of N isotope labelling device, comprising steps of
Step 1: mycelia growth room is placed in 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；
Isotope injection pipe: being connect with mycelia growth room by fixed angle by step 2 with angle correction device, in junction heat Melten gel bonding securely, connects mycelia growth room and the inclination 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: it after the appropriate soil sieving removal root segment in plant growth region, sterilizes through high pressure high humidity and is 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, 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 being 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 isotope is infused 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.
2. arbuscular mycorrhizal fungi according to claim 115The production method of N isotope labelling device, which is characterized in that institute Stating mycelia growth room is 150mlPE vinyon bottle.
3. arbuscular mycorrhizal fungi according to claim 115The production method of N isotope labelling device, which is characterized in that institute Stating filter locking ring is to remove top and screwed bottle cap, mutually close with the plastic bottle port recess of production mycelia growth room Suit.
4. arbuscular mycorrhizal fungi according to claim 115The production method of N isotope labelling device, which is characterized in that institute Stating isotope injection pipe is 1ml plastics aseptic injection pipe, nozzle diameter 0.3mm.
5. arbuscular mycorrhizal fungi according to claim 115The production method of N isotope labelling device, which is characterized in that institute It is 1mm sieve and aperture is 30 μm of nylon wires that state filter, which include aperture,.
6. arbuscular mycorrhizal fungi according to claim 115The production method of N isotope labelling device, which is characterized in that The angle correction equipment material is PVC foam plate, takes angle with angulation tolerance and cutting is bonded, including mycelia growth room is solid Determine area and isotope injection pipe clip plate.
CN201810073591.3A 2018-01-25 2018-01-25 A kind of arbuscular mycorrhizal fungi15N isotope labelling device and production method Active CN108307926B (en)

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Family Cites Families (7)

* Cited by examiner, † Cited by third party
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
CN105675705B (en) * 2016-02-25 2018-12-25 浙江大学 Measure the device and method of amino acid movement law in the 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
CN106834087B (en) * 2017-03-01 2019-02-22 厦门大学 A kind of the AM fungi numerous and functional experiment device of expansion and application method of timing cycle
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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