CN106556667A - A kind of assay method of rhizosphere area soil nitrogen fixation - Google Patents
A kind of assay method of rhizosphere area soil nitrogen fixation Download PDFInfo
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- CN106556667A CN106556667A CN201611049216.2A CN201611049216A CN106556667A CN 106556667 A CN106556667 A CN 106556667A CN 201611049216 A CN201611049216 A CN 201611049216A CN 106556667 A CN106556667 A CN 106556667A
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- G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
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Abstract
The present invention provides a kind of assay method of rhizosphere area soil nitrogen fixation, and sample ground is chosen in ecosystem, in sample ground, lays sample point by the field multiple spot mixing method of sampling, chooses the sample point junior unit with plant；In junior unit, the soil sampling mixing centered on plant, carefully takes the soil body with root system per orientation；The soil body is gently tapped with sterilized soil hammer, and is gently shaken, the bulky grain soil being attached on root system is gently rejected with sterilized soil knife；Then, the rhizosphere soil brush for being close to 1 5mm of root system is entered with sterilized hairbrush, the mixed soil sample of acquirement is immediately placed in into the storage of the couveuse with ice cube, laboratory is transported back；Finally, using soil incubation Kjeldahl's method, determine rhizosphere area's soil nitrogen fixation.The present invention is circulated with important effect for the geobiochemistry for further appreciating that whole rhizosphere area soil nitrogen.
Description
Technical field
The present invention relates to biogeochemistry field, specifically, is related to a kind of measure of rhizosphere area soil nitrogen fixation
Method.
Background technology
Soil nitrogen fixation（Nitrogen fixation）It is biogeochemistry nitrogen（N）The important step of element circulation,
In the presence of soil nitrogen-fixing microorganism, molecular nitrogen is reduced into the process of ammonification and other nitrogen-containing compounds.Contain in the earth's crust
There is few soluble inorganic nitrogen salt, and all biologies are almost required for relying on the nitrogen offer nutrient that diazotroph is fixed in air
Originate and survive, therefore biological nitrogen fixation plays particularly important effect to the nitrogen cycle for maintaining nature.Nitrogen fixation is ground
Study carefully and using the source of manure can be opened up for agricultural, have to maintaining and increasing soil fertility and know meaning.
Primary Actor of the soil nitrogen-fixing microorganism as nitrogen fixation, in rhizosphere area, soil ammonification microbial activity has
Unique characteristic, thus the determination of the assay method of rhizosphere area soil nitrogen fixation, for understanding rhizosphere area soil ecosystem
Have great importance carbon nitrogen balance is adjusted.It is rhizosphere soil to be commonly considered as apart from the shallow soil of root surface l-5mm,
Thus rhizosphere area soil refers to relatively weak that a part of soil scope for directly being affected by root system of plant.Given birth to by root system of plant
The impact of life activity, rhizosphere area root exudates are more enlivened to the material of soil ecosystem, the impact of energy exchange, thus
The soil nitrogen fixation process that rhizosphere area environment has unique physics, chemistry, biological characteristic, the region there is also rhizosphere effect
Should.Therefore, the measure of rhizosphere area soil nitrogen fixation is to further appreciating that soil ecosystem especially rhizosphere area soil nitrogen
Biogeochemical cycle process has important function.
In existing technology, only determine in the whole soil body soil nitrogen fixation intensity and adjusting studying soil ecosystem
Section carbon nitrogen balance, and the determination of the assay method of rhizosphere area soil nitrogen fixation process has not yet to see report.
The content of the invention
It is an object of the invention to provide a kind of assay method of rhizosphere area soil nitrogen fixation.
In order to realize the object of the invention, present invention firstly provides rhizosphere area soil nitrogen fixation process assay method is general
Read, it is desirable to provide a kind of quantitative determination rhizosphere area soil nitrogen fixation method.
The present invention provides a kind of method for determining rhizosphere area's soil nitrogen fixation, chooses 20m × 20m's in ecosystem
Sample ground, in sample ground, lays sample point by the field multiple spot mixing method of sampling, chooses 12 little lists of sample point with plant
Unit；In junior unit centered on plant, by east, south, west, north, the southeast, northeast, southwest, the orientation of northwest 8, in horizontal range
Soil sampling mixing at plant 20cm, 50cm, 80cm, carefully takes with the long 20cm of root system × wide 20cm × depth 50cm per orientation
The soil body；The soil body is gently tapped with sterilized soil hammer, and is gently shaken, gently rejected with sterilized soil knife and be attached to root
The bulky grain soil fastened；Then, the rhizosphere soil brush for being close to root system 1-5mm is entered with sterilized hairbrush and sterilize certainly
Envelope（Wide 17cm × long 24cm）, the mixed soil sample of acquirement is immediately placed in into the storage of the couveuse with ice cube, laboratory is transported back；
Finally, using soil incubation-Kjeldahl's method, determine rhizosphere area's soil nitrogen fixation.
Aforesaid method, the Non-rhizosphere soil sample of acquisition keep low temperature to keep soil fresh in storage process.
Aforesaid method, taking back carries out and completes the early stage culture of nitrogen fixation in 24 h of Non-rhizosphere soil sample of laboratory.
Aforesaid method, adds peptone culture, through the micro- life of fixed nitrogen after culture used in soil incubation method assay method
The nitrogen content that thing is fixed represents the intensity of nitrogen fixation.
Aforesaid method, the nitrogen content fixed through nitrogen-fixing microorganism after cultivating in soil incubation method are specifically determined using kelvin
Nitrogen method is determined.
Aforesaid method, it is described to take pedotheque in east, south, west, north, the southeast, northeast, southwest, the orientation of northwest 8, be
The pedotheque taken at horizontal range plant 20cm, 50cm, 80cm.
Aforesaid method, it is described to take pedotheque volume and be：Long 20cm × wide 20cm × depth 50cm carries root system
The soil body.
Aforesaid method, the valve bag size is：Wide 17cm × long 24cm, material is polyethylene.
Aforesaid method, the rhizosphere soil are the soil for being close to root system 1-5mm.
Described ecosystem includes forest, meadow, wetland or farmland.
Advantages of the present invention effect is as follows：
Soil nitrogen fixation process be biogeochemistry nitrogen cycle and in the presence of soil nitrogen-fixing microorganism by molecular state
Nitrogen is reduced the process of ammonification and other nitrogen-containing compounds, and in existing technology, only determining whole soil includes root system-micro- life
Thing-the litter hybrid system nitrogen fixing capacity studying soil ecosystem adjusting ecosystem carbon, nitrogen balance, and for root
The determination of border area soil nitrogen fixation there is no method intuitively to determine at present.Thus, the present invention is extracted by rejecting rhizosphere area soil
Rhizosphere area soil test rhizosphere area soil nitrogen fixation, for the active rhizosphere area participation for understanding whole material, energy exchange
Geobiochemistry nitrogen cycle has important effect, can incite somebody to action to studying rhizosphere area's soil nitrogen-fixing microorganism for agricultural sector
In air, molecular nitrogen is reduced to ammonia and other nitrogen-containing compounds and is supported using basic data is provided for crops；In addition, right
The pivotal role played in carbon, nitrogen balance in understanding rhizosphere area's biotic environment is important.
Description of the drawings
Schematic diagrams of the Fig. 1 for the soil nitrogen fixation of soybean rhizosphere area.
Schematic diagrams of the Fig. 2 for the soil nitrogen fixation of alfalfa rhizosphere area.
Fig. 3 is the multiple spot mixing method of sampling used in the embodiment of the present invention.
Fig. 4 is that 8 orientation used in the embodiment of the present invention fetch earth the method for sample.
Specific embodiment
Following examples are used to illustrate the present invention, but are not limited to the scope of the present invention.If not specializing, embodiment
In the conventional meanses that are well known to those skilled in the art of technological means used, it is raw materials used to be commercial goods.
The soil incubation method being related in following examples determines soil nitrogen fixation, using addition glucose culture, culture
The nitrogen content of nitrogen-fixing microorganism fixation is measured after one period using Kjeldahl's method.
Embodiment determines farmland（Semen sojae atricolor）, meadow（Alfalfa）The method of rhizosphere area soil nitrogen fixation
First, the sample ground of 20m × 20m is chosen in Semen sojae atricolor, alfalfa ground, in sample ground, mixes sampling side by field multiple spot
Method lays sample point（Fig. 3）, choose 10 sample point junior units with plant；In junior unit centered on plant, by east,
South, west, north, the southeast, northeast, southwest, the orientation of northwest 8（Fig. 4）, the soil sampling at horizontal range plant 20cm, 50cm, 80cm
Mixing, carefully takes the soil body with the long 20cm of root system × wide 20cm × depth 50cm per orientation；Hammered into shape gently with sterilized soil
The soil body is tapped, and is gently shaken, the bulky grain soil being attached on root system is gently rejected with sterilized soil knife；Then, use
The rhizosphere soil brush for being close to root system 1-5mm is entered by sterilized hairbrush, by the mixed soil sample for obtaining immediately
The storage of the couveuse with ice cube is put in, laboratory is transported back；Finally, using soil incubation-Kjeldahl's method, determine rhizosphere area soil
Earth nitrogen fixation.
Aforesaid method, fetching Non-rhizosphere soil sample carries out the early stage culture of nitrogen fixation in 24h.
Aforesaid method, adds glucose culture, through the micro- life of fixed nitrogen after culture used in soil incubation method assay method
The nitrogen content that thing is fixed represents the intensity of nitrogen fixation.
Aforesaid method, the nitrogen content fixed through nitrogen-fixing microorganism after cultivating in soil incubation method adopt Kjeldahl's method
It is measured.
Determination step is as follows：
1）The fresh soil that 100g crosses 2mm sieves is weighed, is positioned in the wide mouthed bottle of 150mL, added 2g glucoses, mix thoroughly（Do not add
Plus glucose is blank process）, add distilled water to water-holding capacity to be 60%, sealed with polyethylene film, incubated at room temperature 30d；
2）Pedotheque after culture is dried immediately, milled 0.149mm sieves, with full nitrogen in Kjeldahl nitrogen determination soil；
3）By adding the difference between the full nitrogen that glucose and blank process are determined, soil nitrogen fixation is calculated strong
Degree.
By calculating, the variation tendency of rhizosphere area soil nitrogen fixation is presented single peak type change in farmland-soybeans,
Summer reaches peak value.Its value is respectively：The soil nitrogen fixation of March rhizosphere area：0.151±0.108 mg/kg；April rhizosphere
Area's soil nitrogen fixation：0.357±0.118 mg/kg；The soil nitrogen fixation of May rhizosphere area：0.421±0.145 mg/kg；
The soil nitrogen fixation of June rhizosphere area：0.825±0.178 mg/kg；The soil nitrogen fixation of July rhizosphere area：0.868±
0.176 mg/kg；The soil nitrogen fixation of 8Yue Fen rhizospheres area：1.134±0.173 mg/kg；9Yue Fen rhizospheres area soil fixed nitrogen is made
With：0.738±0.141 mg/kg；The soil nitrogen fixation of October rhizosphere area：0.237±0.079 mg/kg；November rhizosphere
Area's soil nitrogen fixation：0.113±0.077 mg/kg；Soil nitrogen fixation in soybeans annual rhizosphere area is 0.538 ±
0.363 mg/kg。
Single peak type change is presented in the variation tendency of meadow-alfalfa Yang Didinei rhizospheres area's soil nitrogen fixation,
Summer reaches peak value.Its value is respectively：The soil nitrogen fixation of March rhizosphere area：0.087±0.019 mg/kg；April rhizosphere
Area's soil nitrogen fixation：0.234±0.115 mg/kg；The soil nitrogen fixation of May rhizosphere area：0.582±0.143 mg/kg；
The soil nitrogen fixation of June rhizosphere area：0.804±0.178 mg/kg；The soil nitrogen fixation of July rhizosphere area：1.113±
0.157 mg/kg；The soil nitrogen fixation of 8Yue Fen rhizospheres area：1.142±0.179 mg/kg；9Yue Fen rhizospheres area soil fixed nitrogen is made
With：0.641±0.139 mg/kg；The soil nitrogen fixation of October rhizosphere area：0.186±0.074 mg/kg；November rhizosphere
Area's soil nitrogen fixation：0.069±0.038 mg/kg；The soil nitrogen fixation of alfalfa ground annual rhizosphere area is 0.540 ±
0.421 mg/kg。
It is suitable for wetland or forest land, although, it is above right with a general description of the specific embodiments
The present invention has made detailed description, but on the basis of the present invention, it can be made some modifications or improvements, and this is to art technology
It is obvious for personnel.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to
In the scope of protection of present invention.
Claims (8)
1. a kind of assay method of rhizosphere area soil nitrogen fixation, chooses the sample ground of 20m × 20m, in ecosystem on sample ground
It is interior, sample point is laid by the field multiple spot mixing method of sampling, choose 12 sample point junior units with plant；In junior unit
Centered on plant, by east, south, west, north, the southeast, northeast, southwest, the orientation of northwest 8, horizontal range plant 20cm,
Soil sampling mixing at 50cm, 80cm, carefully takes the soil body with the long 20cm of root system × wide 20cm × depth 50cm per orientation；With
The soil hammer of sterilizing gently taps the soil body, and gently shakes, gently rejected with sterilized soil knife be attached to it is big on root system
Granular soil；Then, the rhizosphere soil brush for being close to root system l-5mm is entered with sterilized hairbrush, will be taken
The mixed soil sample for obtaining is immediately placed in the storage of the couveuse with ice cube, transports laboratory back；Finally, it is fixed using soil incubation-kelvin
Nitrogen method, determines rhizosphere area's soil nitrogen fixation.
2. it is according to claim 1 it is a kind of determine rhizosphere area's soil nitrogen fixation method, it is characterised in that acquisition it is non-
Rhizosphere sampling keeps low temperature to keep soil fresh in storage process.
3. it is according to claim 1 it is a kind of determine rhizosphere area's soil nitrogen fixation method, it is characterised in that take back experiment
The early stage culture of nitrogen fixation is carried out and is completed in 24 h of Non-rhizosphere soil sample of room.
4. it is according to claim 1 it is a kind of determine rhizosphere area's soil nitrogen fixation method, it is characterised in that described soil
Add glucose culture used in earth culture method assay method, after culture, nitrogen fixation is represented through nitrogen-fixing microorganism fixed nitrogen amount
Intensity.
5. it is according to claim 1 it is a kind of determine rhizosphere area's soil nitrogen fixation method, it is characterised in that described survey
Concrete grammar is determined for Kjeldahl's method.
6. it is according to claim 1 it is a kind of determine rhizosphere area's soil nitrogen fixation method, it is characterised in that it is described self-styled
Bag size be：Wide 17cm × long 24cm, material is polyethylene.
7. it is according to claim 1 it is a kind of determine rhizosphere area's soil nitrogen fixation method, it is characterised in that the rhizosphere
Soil is the soil for being close to root system 1-5mm.
8. it is according to claim 1 it is a kind of determine rhizosphere area's soil nitrogen fixation method, it is characterised in that described life
State system includes forest, meadow, wetland or farmland.
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| CN103728433A (en) * | 2013-12-19 | 2014-04-16 | 北京林业大学 | Method for measuring contribution rate of plant root system to nitrification and denitrification of soil |
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Non-Patent Citations (1)
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Application publication date: 20170405 |