CN114080924B - Cultivation device and method for marking woody plant nitrogen stable isotope - Google Patents
Cultivation device and method for marking woody plant nitrogen stable isotope Download PDFInfo
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- CN114080924B CN114080924B CN202111341581.1A CN202111341581A CN114080924B CN 114080924 B CN114080924 B CN 114080924B CN 202111341581 A CN202111341581 A CN 202111341581A CN 114080924 B CN114080924 B CN 114080924B
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 title claims abstract description 142
- 229910052757 nitrogen Inorganic materials 0.000 title claims abstract description 71
- 238000000034 method Methods 0.000 title claims abstract description 30
- 241000196324 Embryophyta Species 0.000 claims abstract description 96
- 239000000463 material Substances 0.000 claims abstract description 40
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 29
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 29
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 29
- 238000001035 drying Methods 0.000 claims abstract description 25
- 238000003306 harvesting Methods 0.000 claims abstract description 9
- 239000002689 soil Substances 0.000 claims description 40
- -1 polypropylene Polymers 0.000 claims description 22
- 239000003337 fertilizer Substances 0.000 claims description 19
- 239000004743 Polypropylene Substances 0.000 claims description 15
- 229920001155 polypropylene Polymers 0.000 claims description 15
- 239000001963 growth medium Substances 0.000 claims description 13
- 239000004576 sand Substances 0.000 claims description 13
- 241000219492 Quercus Species 0.000 claims description 11
- 239000000758 substrate Substances 0.000 claims description 10
- 239000002686 phosphate fertilizer Substances 0.000 claims description 9
- 238000002372 labelling Methods 0.000 claims description 8
- 239000004698 Polyethylene Substances 0.000 claims description 7
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 7
- 229920000573 polyethylene Polymers 0.000 claims description 7
- 229940072033 potash Drugs 0.000 claims description 7
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Substances [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 7
- 235000015320 potassium carbonate Nutrition 0.000 claims description 7
- 230000003014 reinforcing effect Effects 0.000 claims description 3
- 238000011160 research Methods 0.000 description 7
- 239000007787 solid Substances 0.000 description 6
- 230000012010 growth Effects 0.000 description 5
- 238000001948 isotopic labelling Methods 0.000 description 5
- 230000008635 plant growth Effects 0.000 description 5
- 239000004033 plastic Substances 0.000 description 5
- 229920003023 plastic Polymers 0.000 description 5
- 239000002609 medium Substances 0.000 description 4
- 239000000618 nitrogen fertilizer Substances 0.000 description 4
- 239000001166 ammonium sulphate Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000002344 surface layer Substances 0.000 description 3
- 229910021642 ultra pure water Inorganic materials 0.000 description 3
- 239000012498 ultrapure water Substances 0.000 description 3
- 241000209082 Lolium Species 0.000 description 2
- 241000219000 Populus Species 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000003621 irrigation water Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241001480057 Quercus salicina Species 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000004173 biogeochemical cycle Methods 0.000 description 1
- 230000033558 biomineral tissue development Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 210000003298 dental enamel Anatomy 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000002655 kraft paper Substances 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 238000004382 potting Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000000475 sunscreen effect Effects 0.000 description 1
- 239000000516 sunscreening agent Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
- A01G9/021—Pots formed in one piece; Materials used therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
- C05D1/00—Fertilisers containing potassium
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Ecology (AREA)
- Organic Chemistry (AREA)
- Environmental Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Forests & Forestry (AREA)
- Inorganic Chemistry (AREA)
- Pest Control & Pesticides (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Wood Science & Technology (AREA)
- Botany (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention belongs to the technical field of nitrogen stable isotopes, and particularly relates to a cultivation device and a marking method for marking a woody plant nitrogen stable isotope. The invention utilizes a specific cultivation device, adds a cultivation medium into the cultivation device, plants woody plant seedlings, and applies the woody plant seedlings to the root of the seedlings after the woody plant seedlings survive completely 15 N abundance of 30.0atom% 15 N marks ammonium sulfate; and (5) harvesting after 1 year of planting of woody plant seedlings, and drying to obtain the nitrogen stable isotope labeled material. Can obtain a sufficient amount of nitrogen stable isotope labeled test material, and 15 high N abundance, measured 15 The result of the N abundance data is stable and reliable.
Description
Technical Field
The invention belongs to the technical field of nitrogen stable isotopes, and particularly relates to a cultivation device and a marking method for marking a woody plant nitrogen stable isotope.
Background
The nitrogen stable isotope technology is taken as an important technology for researching the terrestrial ecosystem nitrogen bio-geochemical cycle, is widely adopted and applied in domestic and foreign researches, and is considered as the most valuable research tool. In previous studies, stable isotope labeling of a specific type of plant was required for different ecological systems. For example, in agricultural ecosystems and grassland ecosystems, nitrogen stable isotopes @, are shown 15 N) labeled ryegrass (lolium spp.) was used to study the mineralization and assimilation process of soil nitrogen; nitrogen stable isotope label in southwest needle leaf forest ecosystem in siberiaThe noted Populus tremulosa (Populus tremulosa l.) was used to study the main direction of apoptosis nitrogen in soil. Thus, the acquisition of the marker material is particularly critical and important in nitrogen cycling studies in different ecosystems and in different regions. However, the current method for preparing the nitrogen stable isotope labeled plant material is imperfect, and the obtained test material has low abundance, unstable abundance and small quantity, which brings a plurality of inconveniences for subsequent research and analysis.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, and provides a cultivation device and a marking method for marking woody plant nitrogen stable isotopes, which have wide applicability, can obtain a sufficient amount of marking materials with high nitrogen abundance through simple operation, are used for data determination and analysis, and improve the stability and reliability of research.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a cultivation device for woody plant nitrogen stable isotope labeling, which comprises a cultivation unit 8 and a mat net 5; the cultivation unit 8 consists of an upper cultivation element 4 and a lower cultivation element 7 which are vertically arranged, and the mat net 5 is placed at the joint of the upper cultivation element 4 and the lower cultivation element 7;
the bottom of the upper cultivating element 4 is provided with a hole 6.
Preferably, the material of the cultivating unit 8 includes polypropylene or polyethylene; the outer wall of the cultivation unit 8 is provided with reinforcing stripes.
Preferably, the upper cultivation element 4 and the lower cultivation element 7 have the same specification; the diameter of the bottom opening of the upper cultivation element 4 is 10-15 mm.
The invention also provides a method for labeling the woody plant nitrogen stable isotope, which comprises the following steps:
adding a cultivation substrate to the upper cultivation element 4 of the cultivation apparatus;
planting woody plant seedlings on the culture medium, and applying the woody plant seedlings to the root of the seedlings after the woody plant seedlings are all survived 15 N abundance of 30.0atom% 15 N marks ammonium sulfate;
and (5) harvesting after 1 year of planting of woody plant seedlings, and drying to obtain the nitrogen stable isotope labeled material.
Preferably, each woody plant seedling is applied 15 The amount of N-labeled ammonium sulfate is 60.0 to 72.0g.
Preferably, the drying includes a first drying and a second drying which are sequentially performed;
the temperature of the first drying is 105 ℃ and the time is 0.5h; the temperature of the second drying was 80 ℃.
Preferably, the culture medium comprises soil and river sand; the mass ratio of the soil to the river sand is 3:2 to 4:1.
preferably, the culture medium further comprises a base fertilizer; the base fertilizer comprises phosphate fertilizer and potash fertilizer;
the application amount of the phosphate fertilizer is P 2 O 5 Each Kg of cultivation substrate comprises 0.3g of phosphate fertilizer;
the application amount of the potassium fertilizer is K 2 O is calculated, and each Kg of cultivation substrate comprises 0.3g of potash fertilizer.
Preferably, the plant comprises evergreen broadleaf woods dominant vegetation comprising oak.
The invention also provides application of the nitrogen stable isotope labeled material in exploring a nitrogen transfer conversion process, tracking a nitrogen source, tracking a nitrogen direction or revealing a generation mechanism.
The invention provides a cultivation device marked by a woody plant nitrogen stable isotope, which comprises a box body 8 and a mat net 5; the box body consists of an upper cultivation unit 4 and a lower cultivation unit 7 which are vertically arranged, and the cushion net 5 is placed at the joint of the upper cultivation unit 4 and the lower cultivation unit 7; the bottom of the upper cultivation unit 7 is provided with a hole 6. The bottom of the upper cultivation unit 4 of the cultivation device is provided with the holes 6, and the pad net 5 is placed at the joint of the upper cultivation unit 4 and the lower cultivation unit 7, so that soil particles can be prevented from being scattered to the lower cultivation unit at the bottom, and the fixed growth of plant root systems in the cultivation process can be ensured.
Further, the cultivation device of the invention is used for planting woody plant seedlingsAfter the woody plant seedlings in the cultivation device are all survived, applying the woody plant seedlings to the root parts of the seedlings 15 N abundance of 30.0atom% 15 N marks ammonium sulfate; and harvesting leaves, branches and roots after 1 year of plant seedling planting, and drying to obtain the nitrogen stable isotope labeled material. The invention utilizes 15 N marks fertilizer, provides nitrogenous fertilizer for seedling growth, obtains sufficient abundance of marked plant test material, determines the applied in 1 year period according to the abundance of the required plant test material 15 N marks the amount of ammonium sulfate. Research results show that the obtained marked plant test material 15 The abundance of N is stable, and the abundance variation coefficient of the marking materials of different plant parts is low.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments will be briefly described below.
FIG. 1 is a perspective view of a cultivation apparatus provided in example 1;
fig. 2 is a side view of the planting device shown in fig. 1.
Wherein: 1-woody plant seedlings; 2-a cultivation substrate; 3-seedling roots in soil; 4-upper cultivation units; 5-a mat net; 6-hole; 7-a lower cultivation unit; 8-cultivation elements.
Detailed Description
The invention provides a cultivation device for woody plant nitrogen stable isotope labeling. As shown in fig. 1, the cultivation device of the present invention comprises a case 8 and a mat 5; the box body 8 consists of an upper cultivation unit 4 and a lower cultivation unit 7 which are vertically arranged, and the mat net 5 is placed at the joint of the upper cultivation unit 4 and the lower cultivation unit 7;
the bottom of the upper cultivation unit 4 is provided with an opening 6, and the lower cultivation unit 7 is not provided with an opening.
The structure of the cultivation apparatus for nitrogen stable isotope labeling of woody plants according to the present invention will be described with reference to fig. 1 to 2. The cultivation device provided by the invention comprises a box body 8 and a mat net 5. The material of the box body of the invention preferably comprises polypropylene or polyethylene; the outer wall of the box body 8 is preferably provided with reinforcing stripes. The height of the box body 8 is preferably 290-310 mm, and more preferably 300mm; the length of the case 8 is preferably 480 to 660mm, more preferably 500 to 600mm, and even more preferably 550mm; the width of the case 8 is preferably 270 to 500mm, more preferably 300 to 450mm, and even more preferably 350 to 400mm; the wall thickness of the case 8 is preferably 2.0 to 4.0mm, more preferably 2.5 to 3.5mm, and still more preferably 3.0mm. The box body 8 can resist high pressure, and the invention has no special requirement on the structure of the reinforced stripes on the outer wall of the box body 8.
The cultivation element 8 of the invention consists of an upper cultivation unit 4 and a lower cultivation unit 7 which are vertically arranged, wherein a hole 6 is arranged at the bottom of the upper cultivation unit 4. The height of the upper cultivation unit 4 is preferably 145-155 mm, more preferably 150mm; the length of the upper cultivating unit 4 is preferably 480 to 660mm, more preferably 500 to 600mm, and even more preferably 550mm; the width of the upper cultivating unit 4 is preferably 270 to 500mm, more preferably 300 to 450mm, and even more preferably 350 to 400mm; the wall thickness of the upper cultivation element 4 is preferably 2.0 to 4.0mm, more preferably 2.5 to 3.5mm, and even more preferably 3.0mm. The diameter of the bottom opening 6 of the upper cultivating unit 4 is preferably 10-15 mm, more preferably 12-14 mm. The number of the holes 6 at the bottom of the upper cultivating unit 4 is preferably 9 to 12, more preferably 10. The invention has no special requirement on the position of the holes 6 at the bottom of the upper cultivation unit 4, ensures the diameter and the number of the holes, and preferably uniformly opens the holes. The upper cultivation unit provided by the invention has the advantages that the soil ventilation can be increased, the drainage is facilitated, and the normal growth of plants is ensured.
The height of the lower cultivation unit 7 is preferably 145-155 mm, more preferably 150mm; the length of the lower cultivation unit 7 is preferably 480 to 660mm, more preferably 500 to 600mm, and even more preferably 550mm; the width of the lower cultivation unit 7 is preferably 270 to 500mm, more preferably 300 to 450mm, and even more preferably 350 to 400mm; the wall thickness of the lower cultivation element 7 is preferably 2.0 to 4.0mm, more preferably 2.5 to 3.5mm, and even more preferably 3.0mm. The present invention preferably has the same specifications for the upper cultivation unit 4 and the lower cultivation unit 7.
The invention places the mat 5 at the junction of the upper cultivation unit 4 and the lower cultivation unit 7. The mat web 5 according to the invention is preferably a 6-8 needle mat web. The width of the mat web 5 according to the present invention is preferably 400 to 410mm, more preferably 405mm; the length of the mat web 5 is preferably 660 to 680mm, more preferably 665 to 675mm. The invention has no strict requirements on the color and the material of the pad net 5, and the color of the pad net 5 is preferably black; the mat 5 is preferably made of polyethylene, polypropylene or other materials which do not contain toxic or harmful substances, and which do not pollute the soil and do not affect plant growth.
In the invention, the upper cultivation unit 4 cultures woody plant seedlings, ensures the fixed growth of plant root systems in the cultivation process, and the lower cultivation unit 7 can collect irrigation water leached by soil in the cultivation process of the woody plant seedlings, so that the irrigation water is recycled, the repeatability is good, the applicability is wide, and the cultivation cost is reduced. According to the invention, the mat net 5 is placed at the joint of the upper cultivation unit 4 and the lower cultivation unit 7, so that the fixed growth of plant root systems and the phenomenon that soil particles are scattered to the bottom of the lower cultivation unit solution collecting box in the cultivation process can be ensured.
The invention uses the cultivation device to label the woody plant nitrogen stable isotope to obtain the nitrogen stable isotope labeled material, and the labeling method comprises the following steps:
adding a culture medium to an upper culture unit 4 of the culture device;
planting woody plant seedlings on the culture medium, and applying the woody plant seedlings to the root of the seedlings after the woody plant seedlings are all survived 15 N abundance of 30.0atom% 15 N marks ammonium sulfate;
and (5) harvesting after 1 year of planting of woody plant seedlings, and drying to obtain the nitrogen stable isotope labeled material.
The present invention adds a culture medium to the upper cultivation element 4 of the cultivation apparatus. The culture medium of the invention preferably comprises soil, river sand and base fertilizer. The soil of the invention is preferably soil at a position 0-20 cm away from the surface layer, more preferably soil at a position 5-15 cm away from the surface layer, and even more preferably soil at a position 8-12 cm away from the surface layer; and is also provided withThe nitrogen content of the soil is preferably<0.5%. The mass ratio of the soil to the river sand is preferably 3:2 to 4:1, more preferably 2:1 to 3:1, more preferably 7:3. The type of the soil is not particularly limited, and red soil is preferable in the examples. The mass of the base fertilizer in the culture medium of the present invention is preferably 0.5 to 0.6g, more preferably 0.6g, based on 1kg of the total mass of the soil and the river sand. The base fertilizer is preferably phosphate fertilizer and potash fertilizer, and the phosphate fertilizer adopts P 2 O 5 Counting the potassium fertilizer by K 2 The mass ratio of the phosphate fertilizer to the potash fertilizer is preferably 2:1 to 1:1, more preferably 1:1.
After a cultivation medium is added into a cultivation device, the invention plants woody plant seedlings on the cultivation medium, and after the woody plant seedlings survive completely, the seedlings are applied to the roots of the seedlings 15 N abundance of 30.0atom% 15 N marks ammonium sulfate. The plant height of the woody plant seedling is preferably 9.0 cm-11.0 cm, and more preferably 9.5 cm-10.5 cm; the dry weight of the woody plant seedlings (excluding the dry weight of the seedling roots) is preferably 1 to 2g, more preferably 1.5g. The woody plant seedling of the invention is preferably a 1 year old woody plant seedling with regular growth and no lateral branches, and the woody plant preferably comprises evergreen broad-leaved forest dominant vegetation, and more preferably comprises oak. The number of seedlings of the woody plant according to the present invention is conventionally selected according to the specifications of the cultivation apparatus, and preferably, it is planted according to the specifications of 3 (rows) ×6 (columns).
The pair of the invention 15 N abundance of 30.0atom% 15 The source of the N-labeled ammonium sulfate is not particularly limited and is commercially available by conventional means.
When woody plant seedlings are planted, the seedlings are uniformly transplanted and planted in a cultivation device, soil moisture is fully poured, and a rain-proof shed is installed. And (5) watering regularly, removing weeds in the cultivation device and checking the growth condition of seedlings. The present invention preferably embeds the removed weeds in the soil of the cultivation device.
After the woody plant seedlings of the invention survive completely, the root parts of the seedlings are applied 15 N abundance of 30.0atom% 15 N marks ammonium sulfate. Application of the inventionThe said 15 The amount of N-labeled ammonium sulfate is preferably 3 to 5g, more preferably 4g, per woody plant seedling. The invention applies the following 15 In the case of N-labelled ammonium sulphate, it is preferably applied in stages, more preferably comprising: 1 application of 30.0atom% abundance every 1 month in spring and summer 15 N-labeled ammonium sulfate was applied 1 time every two months in autumn and winter with an abundance of 30.0atom% 15 N-labeled ammonium sulfate, more preferably, the abundance ratio is 30.0atom% per application 15 The amount of N-labelled ammonium sulphate was the same. The invention is described in 15 N abundance of 30.0atom% 15 The application amount of N marked ammonium sulfate is equal to 1.2-1.4 g/Kg of nitrogen fertilizer. In the cultivation process, if the seedlings which do not survive appear, the seedlings need to be planted in a supplementary mode in time.
In the invention, the ammonium sulfate is preferably ammonium sulfate solid, and more preferably pure ammonium sulfate solid with purity of more than or equal to 99.5%. In particular embodiments of the invention, the applied ammonium sulfate solids are preferably purchased from Shanghai chemical institute.
The invention preferably applies the seedlings to the roots of the seedlings after the seedlings survive 15 N abundance of 30.0atom% 15 N-labelled ammonium sulphate is further preferably applied 1 month after seedling transplanting. The invention transplants the seedling for 1 month to ensure the survival of the seedling.
Woody plant seedlings were grown for 1 year and then the woody plant seedlings were harvested for leaves, branches and roots. In the harvesting process, the invention preferably measures the plant height in the cultivation device, cuts off the plant root, places the branch and leaf parts of the plant in the net bag, and pours out the residual root and soil from the cultivation device. Classifying all plant materials according to leaves, branches and roots to obtain the leaves, branches and roots of the cultivated woody plants.
The invention harvests woody plant seedlings, and after obtaining the leaves, branches and roots of the woody plant after cultivation, the leaves, branches and roots of the woody plant after cultivation are dried to obtain the nitrogen stable isotope labeled material. The drying according to the present invention preferably includes a first drying and a second drying which are sequentially performed; the temperature of the first drying is preferably 100 to 105 ℃, and more preferably 105 ℃; the time for the first drying is preferably 0.4 to 0.6 hours, more preferably 0.5 hours. The temperature of the second drying according to the present invention is preferably 75 to 85 ℃, and more preferably 80 ℃. The invention has no strict requirement on the time of the second drying, and the leaves, branches and roots of the cultivated woody plants are dried to constant weight.
The invention also provides application of the nitrogen stable isotope labeled material obtained by the method in exploring a nitrogen transfer and conversion process, tracking a nitrogen source, tracking a nitrogen destination or revealing a generation mechanism. The nitrogen stable isotope labeled material obtained by the invention has large quantity, 15 high N abundance, measured 15 The result of N abundance data is stable and reliable, and the N abundance data is tracked 15 N can explore the transfer and conversion process of nitrogen, trace the source of nitrogen, trace the direction of nitrogen or reveal the occurrence mechanism.
For further explanation of the present invention, the method for labeling nitrogen stable isotopes of woody plants provided by the present invention will be described in detail with reference to the accompanying drawings and examples, which should not be construed as limiting the scope of the invention.
Example 1
Referring to fig. 1 and 2, the present embodiment provides a cultivation device for nitrogen stable isotope labeling of woody plants, which is composed of a green oak seedling 1, a cultivation substrate 2, a seedling root 3 in soil, an upper cultivation element 4 made of polypropylene, a mat 5, and a lower cultivation element 7 made of polypropylene.
The oak seedlings 1 are planted in the polypropylene upper cultivating element 4, the polypropylene upper cultivating element 4 is provided with a cultivating substrate 2 with specific configuration, holes 6 are formed in the bottom of the polypropylene upper cultivating element 4, a black pad net 5 is placed at the bottom of the polypropylene upper cultivating element 4, the polypropylene upper cultivating element 4 and a polypropylene lower cultivating element 7 are placed in a stacked mode to be the same whole, and holes are not formed in the bottom of the polypropylene lower cultivating element 7.
Example 2
1. Preparation of a cultivation substrate:
materials and tools: spades, hammers, typical red soil, river sand, 20 mesh screens, phosphate fertilizer and potash fertilizer.
When preparing the culture medium, selecting a forest in the Changsha county of Changsha city in Hunan provinceRed soil (nitrogen content 4.0g/kg, phosphorus content 0.15 g/kg) of park evergreen broadleaf woodland. When sampling, firstly removing the ground falling object, digging the surface soil (0-20 cm) by a shovel, and taking about 20.0kg of fresh soil. The soil was brought back to the laboratory and naturally air-dried in the shade for 1 week, after which visible stones and root systems were removed, and the hammers crushed the soil and passed through a 20 mesh screen. 14.0kg of air-dried soil sample (containing 4.0g/kg of N and 0.15g/kg of P), 6.0kg of river sand (the ratio is 7:3) and base fertilizer (NaH) are taken 2 PO 4 ，0.3g P 2 O 5 /kg；KCl，0.3g K 2 O/kg), the air-dried soil, river sand and a base fertilizer shovel are fully and uniformly mixed on the ground, and the cultivation substrate is obtained.
2. Seedling cultivation:
materials and tools: the subtropical land ecosystem is characterized in that the evergreen broad-leaved forest dominant vegetation type, namely, the oak seedlings, a cultivation medium and two cultivation devices are adopted, wherein one bottom part of the cultivation medium is uniformly perforated; a black mat net; watering can; a rain-proof shed and a bracket.
In seedling cultivation, 1 year-old Qinggang oak seedlings with uniform growth and no lateral branches and about 10.0cm in plant height and about 1.5g (without roots) in plant dry weight are purchased in local nursery. 18 seedlings of Quercus cyclobalanopsis are uniformly transplanted into the cultivation device described in example 1 in 3-7.2017 according to the arrangement of 3 (rows) ×6 (columns), and soil moisture is fully filled with watering pots. And placing the cultivation soil box above the non-perforated plastic box, and stacking and placing. And then, the box body is moved to a fixed place, and a rain-proof shed is installed. Regularly watering, removing weeds in the cultivation device, checking the growth condition of seedlings, and timely supplementing and planting the non-surviving seedlings, wherein the removed weeds are buried in the soil of the cultivation device.
3. Application of 15 N-labeled ammonium sulfate:
materials and tools: 15 ammonium sulfate solids with an N abundance of 30.0atom% (purchased from Shanghai chemical institute), precision electronic analytical balances (ME 204, mertrele tolidox), measuring cylinders, ultrapure water, beakers or enamel cups, glass rods, plastic bottles and syringes.
After the seedlings of the Quercus salicina in the cultivation device are all survived for 1 month, accurately weighing the weight of about 7.02g in a laboratory, 15 the N abundance is 30.0atom% 15 N-labeled ammonium sulfate (. About.) 15 NH 4 ) 2 SO 4 ) Solid and formulated as 180.0mL solution. All the solutions were transferred to 250mL polypropylene (PP) plastic bottles, brought to field potting, and applied quantitatively and evenly to the roots of seedlings using a syringe, with 10.0mL of solution applied per seedling. Thereafter, a labeled ammonium sulfate solution having an abundance of 30.0atom% was applied 1 time every two months in spring and summer and every two months in autumn and winter. For 1 year of planting and nitrogen fertilizer application, the total application amount of ammonium sulfate solids was about 63.20g, which corresponds to 1.3g/kg of applied nitrogen fertilizer.
4. And (3) harvesting plants:
materials and tools: tape measure, scissors, nylon net bag, polyethylene plastic bag, ultrapure water, kraft paper envelope and constant temperature drying oven.
Young oak seedlings are harvested 3 months and 7 days in 2018. When harvesting, firstly, the plant height in the cultivation basin is measured by using a rolling ruler, then, the plant is sheared from the root of the plant by using scissors, the branch and leaf parts of the plant are placed in a net bag and brought back to a laboratory, and the residual root and soil are poured out from the cultivation basin. The plant root system attached to the black sun screen is cleaned, and put into a plastic bag to be brought back to a laboratory, and the root muddy sand is carefully cleaned by tap water. Cleaning all plant parts with ultrapure water again, air-drying, and weighing fresh weight.
After the oak plant grows for 1 year, the average plant height of each oak plant is 870mm (+ -92 mm), the average leaf fresh weight is 19.20g (+ -2.62 g), the branch fresh weight is 19.00g (+ -2.60 g), and the root fresh weight is 21.47g (+ -6.31 g). And (3) classifying all plant materials according to leaves, branches and roots, fully and uniformly mixing, filling the plant materials into envelopes, placing the envelopes in a constant-temperature drying oven, deactivating enzymes at 105 ℃ for 0.5h, and drying the plants at 80 ℃ until the weight is constant, thus obtaining the nitrogen stable isotope labeled material.
Example 3
The difference from example 2 is that in step 1, 12.0kg of air-dried soil sample and 8.0kg of river sand (the ratio is 3:2) are taken when preparing the culture medium.
Example 4
The procedure is as in example 2, except that in step 1, 16.0kg of air-dried soil sample and 4.0kg of river sand (the ratio is 4:1) are taken when the cultivation substrate is prepared.
Test example 1
The nitrogen-stable isotope-labeled material obtained in example 2 was analyzed and detected by using an isotope mass spectrometer, a precision electronic analytical balance, a pulverizer, an 80 mesh screen, and a polyethylene self-sealing bag.
The detection method is that the example 2 is dried 15 The N-labeled leaves, branches and roots were again accurately weighed, a portion of the plant sample was uniformly taken according to the quartering method, pulverized into fine powder by a pulverizer, and passed through an 80-mesh screen. The crushed samples are stored in polyethylene self-sealing bags and then sent to an isotope mass spectrometer analysis laboratory for measuring and analyzing each part of plants 15 N abundance (atomic%) and nitrogen content, and the data results are shown in table 1.
TABLE 1 Nitrogen stable isotope labeled materials of each fraction 15 N abundance and Nitrogen content
As can be seen from Table 1, the nitrogen stable isotope labeled leaves in the nitrogen stable isotope labeled material obtained by the method for labeling nitrogen stable isotopes of woody plants of the present invention 15 The N abundance is 16.39atom, and the nitrogen content is 20.62g/kg; nitrogen stable isotope labeled branch 15 The N abundance is 15.93%, and the nitrogen content is 11.70g/kg; nitrogen stable isotope labeled root 15 The N abundance is 15.28atom, and the nitrogen content is 15.50g/kg. The nitrogen stable isotope labeled material obtained by the invention 15 The N abundance is high, nitrogen stable marking conditions of different plant parts are measured, the abundance variation coefficient of the marking material is low, and the abundance and the content are stable.
As can be seen from the above examples, the method for labeling woody plant nitrogen stable isotopes provided by the present invention not only can obtain enough labeling materials (about 600.0g of total dry weight), but also the nitrogen abundance of the labeling materials is about 15.28-16.69 atom%, and the nitrogen content is 11.7020.62g/kg， 15 The method for marking the nitrogen stable isotope of the woody plant has the advantages of high abundance, stable abundance and stable content, has referential property in the research of different ecological systems and environmental nitrogen circulation, and can be used in the research of the nitrogen circulation of the land ecological system in multiple fields.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (6)
1. A method for labeling a nitrogen stable isotope of a woody plant, comprising the steps of:
adding a cultivation substrate into a cultivation device;
planting woody plant seedlings on the culture medium, wherein the woody plant seedlings are 1-year-old woody plant seedlings which grow symmetrically and have no lateral branches, and applying the woody plant seedlings to the root of the seedlings after the woody plant seedlings survive 15 N abundance of 30.0atom% 15 N-labeled ammonium sulfate solution applied 1 time every 1 month in spring and summer with abundance of 30.0atom% 15 N-labeled ammonium sulfate was applied 1 time every two months in autumn and winter with an abundance of 30.0atom% 15 N marks ammonium sulfate; wherein each woody plant seedling is applied 15 The amount of N-labeled ammonium sulfate is 60.0-72.0 g;
harvesting after planting woody plant seedlings for 1 year, and drying to obtain a nitrogen stable isotope labeled material;
the culture medium consists of soil, river sand and base fertilizer; the mass ratio of the soil to the river sand is 3:2-4:1; the base fertilizer is phosphate fertilizer and potash fertilizer; the mass ratio of the phosphate fertilizer to the potash fertilizer is 2:1-1:1;
the cultivation device consists of a cultivation unit (8) and a mat net (5); the cultivation unit (8) consists of an upper cultivation element (4) and a lower cultivation element (7) which are vertically arranged, and the mat net (5) is placed at the joint of the upper cultivation element (4) and the lower cultivation element (7);
the bottom of the upper cultivation element (4) is provided with a hole (6);
the adding position of the culture medium is an upper culture element (4).
2. A method according to claim 1, characterized in that the material of the cultivation element (8) comprises polypropylene or polyethylene; the outer wall of the cultivation unit (8) is provided with reinforcing stripes.
3. A method according to claim 1, characterized in that the upper (4) and lower (7) cultivation elements are of the same specification; the diameter of the bottom opening of the upper cultivation element (4) is 10-15 mm.
4. The method of claim 1, wherein the drying comprises a first drying and a second drying performed sequentially;
the temperature of the first drying is 105 ℃, the time is 0.5h, and the temperature of the second drying is 80 ℃.
5. The method of claim 1, wherein the plant comprises evergreen broadleaf forest-dominant vegetation comprising quercus cyclobalanopsis.
6. Use of a nitrogen stable isotope labelled material obtainable by the method of any one of claims 1 to 5 for exploring the transfer process of nitrogen, tracking the source of nitrogen, tracking the direction of nitrogen or revealing the mechanism of occurrence.
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