CN103636416A - Application of MgCl2 in plant nitrate nitrogen absorption promoting - Google Patents
Application of MgCl2 in plant nitrate nitrogen absorption promoting Download PDFInfo
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- CN103636416A CN103636416A CN201310630601.6A CN201310630601A CN103636416A CN 103636416 A CN103636416 A CN 103636416A CN 201310630601 A CN201310630601 A CN 201310630601A CN 103636416 A CN103636416 A CN 103636416A
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- Prior art keywords
- mgcl
- india canna
- nitrate
- plant
- processing
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- MMDJDBSEMBIJBB-UHFFFAOYSA-N [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] Chemical compound [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] MMDJDBSEMBIJBB-UHFFFAOYSA-N 0.000 title claims abstract description 26
- 238000010521 absorption reaction Methods 0.000 title abstract description 28
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 title abstract description 16
- 229910001629 magnesium chloride Inorganic materials 0.000 title abstract description 8
- 230000001737 promoting effect Effects 0.000 title abstract description 4
- 239000002351 wastewater Substances 0.000 claims abstract description 14
- 239000000126 substance Substances 0.000 claims abstract description 6
- 229910002651 NO3 Inorganic materials 0.000 claims description 28
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 claims description 28
- 230000012010 growth Effects 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 46
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 23
- 102000004190 Enzymes Human genes 0.000 abstract description 11
- 108090000790 Enzymes Proteins 0.000 abstract description 11
- 108090000913 Nitrate Reductases Proteins 0.000 abstract description 11
- 102000005396 glutamine synthetase Human genes 0.000 abstract description 9
- 108020002326 glutamine synthetase Proteins 0.000 abstract description 9
- 230000004060 metabolic process Effects 0.000 abstract description 4
- 102000015176 Proton-Translocating ATPases Human genes 0.000 abstract 1
- 108010039518 Proton-Translocating ATPases Proteins 0.000 abstract 1
- 230000002255 enzymatic effect Effects 0.000 abstract 1
- 238000007781 pre-processing Methods 0.000 abstract 1
- 235000005273 Canna coccinea Nutrition 0.000 description 109
- 241000234587 Canna Species 0.000 description 108
- 238000012545 processing Methods 0.000 description 56
- 241000196324 Embryophyta Species 0.000 description 36
- 230000000694 effects Effects 0.000 description 35
- 238000000034 method Methods 0.000 description 32
- 239000007788 liquid Substances 0.000 description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- 235000018102 proteins Nutrition 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 14
- 108090000623 proteins and genes Proteins 0.000 description 14
- 210000000170 cell membrane Anatomy 0.000 description 11
- 230000008569 process Effects 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000000284 extract Substances 0.000 description 8
- 239000002689 soil Substances 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 7
- 238000010586 diagram Methods 0.000 description 7
- 239000011777 magnesium Substances 0.000 description 7
- 229910052749 magnesium Inorganic materials 0.000 description 7
- 239000012528 membrane Substances 0.000 description 6
- 150000002823 nitrates Chemical class 0.000 description 6
- 238000001556 precipitation Methods 0.000 description 6
- 230000008859 change Effects 0.000 description 5
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 238000004065 wastewater treatment Methods 0.000 description 5
- YXIWHUQXZSMYRE-UHFFFAOYSA-N 1,3-benzothiazole-2-thiol Chemical compound C1=CC=C2SC(S)=NC2=C1 YXIWHUQXZSMYRE-UHFFFAOYSA-N 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 4
- 101001091385 Homo sapiens Kallikrein-6 Proteins 0.000 description 4
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- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- DPKHZNPWBDQZCN-UHFFFAOYSA-N acridine orange free base Chemical compound C1=CC(N(C)C)=CC2=NC3=CC(N(C)C)=CC=C3C=C21 DPKHZNPWBDQZCN-UHFFFAOYSA-N 0.000 description 3
- DZBUGLKDJFMEHC-UHFFFAOYSA-N benzoquinolinylidene Natural products C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 3
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- DEFVIWRASFVYLL-UHFFFAOYSA-N ethylene glycol bis(2-aminoethyl)tetraacetic acid Chemical compound OC(=O)CN(CC(O)=O)CCOCCOCCN(CC(O)=O)CC(O)=O DEFVIWRASFVYLL-UHFFFAOYSA-N 0.000 description 3
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- RUFPHBVGCFYCNW-UHFFFAOYSA-N 1-naphthylamine Chemical compound C1=CC=C2C(N)=CC=CC2=C1 RUFPHBVGCFYCNW-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 108090000854 Oxidoreductases Proteins 0.000 description 2
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- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical class [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
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- 230000031700 light absorption Effects 0.000 description 2
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- 235000013923 monosodium glutamate Nutrition 0.000 description 2
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 2
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- FDDDEECHVMSUSB-UHFFFAOYSA-N sulfanilamide Chemical compound NC1=CC=C(S(N)(=O)=O)C=C1 FDDDEECHVMSUSB-UHFFFAOYSA-N 0.000 description 2
- 229940124530 sulfonamide Drugs 0.000 description 2
- 241000234588 Canna indica Species 0.000 description 1
- 241000234586 Cannaceae Species 0.000 description 1
- WTDHULULXKLSOZ-UHFFFAOYSA-N Hydroxylamine hydrochloride Chemical class Cl.ON WTDHULULXKLSOZ-UHFFFAOYSA-N 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 108010025915 Nitrite Reductases Proteins 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- JVMRPSJZNHXORP-UHFFFAOYSA-N ON=O.ON=O.ON=O.N Chemical compound ON=O.ON=O.ON=O.N JVMRPSJZNHXORP-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 1
- 229910021502 aluminium hydroxide Inorganic materials 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229930002875 chlorophyll Natural products 0.000 description 1
- 235000019804 chlorophyll Nutrition 0.000 description 1
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 1
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- 235000013922 glutamic acid Nutrition 0.000 description 1
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- 229910052751 metal Inorganic materials 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 239000000618 nitrogen fertilizer Substances 0.000 description 1
- -1 nitrogen-containing compound Chemical class 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
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- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- RMJPDRUNCDRUQC-MCDZGGTQSA-M sodium;[[[(2r,3s,4r,5r)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-hydroxyphosphoryl] hydrogen phosphate Chemical compound [Na+].C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)([O-])=O)[C@@H](O)[C@H]1O RMJPDRUNCDRUQC-MCDZGGTQSA-M 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a novel application of MgCl2, namely an application of MgCl2 in plant nitrate nitrogen absorption promoting. During using, an MgCl2 solution with the amount-of-substance concentration of 130-180 micro-mol/L is used for preprocessing plants for 6-8 hours, then the plants are placed into farmland waste water with nitrate nitrogen, and the growing index of the plants is observed; and test results show that the absorption capacity on nitrate nitrogen in the farmland waste water of the plants processed with MgCl2 is enhanced, the enzymatic activity of plasmalemma H+-ATPase and nitrogen metabolism relevant enzymes such as nitrate reductase and glutamine synthetase is enhanced, and MgCl2 can promote nitrogen absorption assimilation of the plants.
Description
Technical field
The invention belongs to the field that promotes that plant nitrate nitrogen absorbs, be specifically related to inorganic chemical reagent MgCl
2new purposes in promoting the absorption of plant nitrate nitrogen.
Background technology
In recent years, the development of China's agricultural has been accelerated in the maturation of establishment planting technology and popularization.The area of the polyhouse-based agriculture of China production at present constantly expands, and within 2011, domestic facilities horticulture area under cultivation surpasses 2,400,000 hm
2, rank first in the world.But the environmental problem that greenhouse-environment produces is in process of production more serious than open-air atmosphere.Facilities vegetable, the floriculture cycle is short, cropping index is high, pattern of farming is single, adds that facility cultivation soil covers throughout the year to can not get rain-out, generally after using 3-5, all secondary salinization in various degree can occur.The salinity anion of facility soil is mainly NO
3 -, be the Main Barrier Factors that facilities vegetable is produced.It is the farmland that is soaked in water that soil after can not cultivating goes saliferous mode, further makes the waste water that contains a large amount of nitrogens enter rivers and lakes, further causes the eutrophication of water body.Above-mentioned resource and the ecological environment problem of both having related to, also closely bound up with agriculture sustainable development, be problem demanding prompt solution during current facilities vegetable is produced.Therefore, invent a kind of efficient, easy, cheap and easy received plant nitrate nitrogen sorbefacient significant.
India canna (
canna indica) be Cannaceae corm kind flower plant, perennial vertical herbage, happiness high temperature and sunlight, flowering stage is long, and strong adaptability has higher ornamental value.In recent years along with the development of Wastewater Treatment by Constructed Wetlands technology, people show its research, and India canna root system is more flourishing, better to the removal effect of TN, TP, COD, the conventional plant that has become eutrophied water treatment, has wide application and promotion prospect.
Magnesium has participated in whole process of growths of all life body, and the fourth-largest of plant must nutritive element afterwards by clay fertilizer scholar, to be regarded as nitrogen, phosphorus, potassium.Known magnesium is chlorophyllous important component part, is also metallic element unique in chlorophyll.Magnesium is fundamental and the most active part of various enzymes, has participated in widely the metabolic processes of plant, is that plant forms starch and the indispensable part of protein.Magnesium has become the hot issue of international agronomy Jie He clay fertilizer circle research on the impact of plant.Existing research shows, plant is a very complicated process to the absorption of nitrogen, assimilation.Nitrate nitrogen is one of inorganic nitrogen form of mainly absorbing of plant, after root system of plant absorbs, also needs could be assimilated by plant through a series of effects such as nitrate reductase, nitrite reductase, glutamine synthelase, glutamate synthetases.Plasma membrane H
+-ATPase is functional protein important on plant cell membrane, is called as " the dominant force enzyme " of plant vital activity process, can set up cross-film proton electromotive force, promotes the transmembrane transport of ion and nutriment.Nitrate reductase (NR) can be nitrite by the nitrate transformation in body, is the first step of nitrate assimilation process, is the rate-limiting enzyme of nitrogen metabolism.Glutamine synthelase (GS) is one of key enzyme of plant nitrogen metabolism, and it is catalysis glutamic acid (Glu) and NH in nitrogen assimilation circulation
3condensation forms glutamine (Gln), and the metabolic pathway of involved in plant nitrogen-containing compound.At present, about the existing greater advance of research of nitrogen absorbed by plant element mechanism, but also do not find the method that directly effectively improves nitrogen absorbed by plant element.People or mainly improves the absorption of plant to nitrogen by increasing the usage amount of nitrogenous fertilizer in actual production, but produce effects very limited and caused the problems referred to above.
Summary of the invention
The object of this invention is to provide a kind of MgCl
2new purposes, as plant nitrate nitrogen sorbefacient, i.e. MgCl
2in the application as in the absorption of the efficient Nitrate in Plants of promotion fast, assimilation.
In order to realize above-mentioned purpose of the present invention, technical scheme of the present invention is as follows:
(1) preparation amount of substance concentration is the MgCl of 130-180 μ mol/L
2solution;
(2) the India canna seedling of plant height 30-40cm in wetland soil is placed in to above-mentioned steps (1) MgCl
2in solution, India Canna Rhizome is carried out to the pretreatment of 6-8h;
(3) pretreated plant is placed in the agricultural wastewater of variable concentrations nitrate nitrogen, and observation of plant is to the absorption rate of nitrate nitrogen and measure the activity that nitrate nitrogen absorbs, assimilates key enzyme.
MgCl provided by the invention
2promoter is easy to use, and cost is lower; This promoter has significantly increased absorption and the assimilation of Nitrate in Plants.The present invention has opened up and has promoted that plant nitrogen absorbs, the new way of assimilation, contributes to scientific worker to MgCl
2the research of the molecule mechanism of stimulating plant growth holds out broad prospects in agricultural production.
Beneficial effect of the present invention: the MgCl of promotion plant nitrogen assimilation of the present invention
2promoter, has the feature low, simple to operate, efficiency is high that drops into, at normal temperatures, and MgCl
2be the promoter that more satisfactory promotion plant nitrate nitrogen absorbs, magnesium chloride pretreatment can improve H on India canna cell membrane
+the activity of the Nitrogen Absorption such as-ATPase, nitrate reductase and glutamine synthelase and assimilation relevant enzyme, absorption and the assimilatory efficiency of raising nitrogen; Contribute to reduce the problems such as the soil salinization that cause because of excessive fertilization, significant to agriculture sustainable development.
Accompanying drawing explanation
Fig. 1 is that the present invention adds promoter 150 μ mol/L MgCl
2process the purify liquid waste result schematic diagram of efficiency of India canna before and after 8h, wherein A figure is without MgCl
2process the absorption result of India Canna Rhizome to nitrate; B figure is through MgCl
2the absorption result of India Canna Rhizome to nitrate after processing;
Fig. 2 is that the present invention adds captax gCl
2to India canna plasma membrane H
+-ATPase activity affect result schematic diagram, wherein A figure is MgCl
2plasma membrane H in India Canna Rhizome before and after processing
+the active result of-ATPase, CK is without MgCl
2the India canna of processing; B figure is MgCl
2plasma membrane H in India canna leaf before and after processing
+the active result of-ATPase, CK is without MgCl
2the India canna of processing;
Fig. 3 is that the present invention adds captax gCl
2to India canna root plasma membrane H
+-pump activity influence result schematic diagram, wherein A figure is not for passing through MgCl
2the control group of processing, represents with CK; B figure is through MgCl
2the experimental group of processing;
Fig. 4 is that the present invention adds captax gCl
2the result schematic diagram that affects on India canna nitrate reductase activity, wherein A figure is MgCl
2nitrate reductase activity result in India Canna Rhizome before and after processing, CK is without MgCl
2the India canna of processing; B figure is MgCl
2nitrate reductase activity result in India canna leaf before and after processing, CK is without MgCl
2the India canna of processing;
Fig. 5 is that the present invention adds captax gCl
2the result schematic diagram that affects on India canna activity of glutamine synthetase, wherein A figure is MgCl
2activity of glutamine synthetase result in India Canna Rhizome before and after processing, CK is without MgCl
2the India canna of processing; B figure is MgCl
2activity of glutamine synthetase result in India canna leaf before and after processing, CK is without MgCl
2the India canna of processing;
Fig. 6 is that the present invention adds promoter 130 μ mol/L MgCl
2process the purify liquid waste result schematic diagram of efficiency of India canna before and after 6h, wherein A figure is not for passing through MgCl
2process the absorption result of India Canna Rhizome to nitrate; B figure is through MgCl
2the absorption result of India Canna Rhizome to nitrate after processing.
Fig. 7 is that the present invention adds promoter 180 μ mol/L MgCl
2process the purify liquid waste result schematic diagram of efficiency of India canna before and after 7h, wherein A figure is not for passing through MgCl
2process the absorption result of India Canna Rhizome to nitrate; B figure is through MgCl
2the absorption result of India Canna Rhizome to nitrate after processing.
Embodiment
Below by drawings and Examples, the present invention is described in further detail, but protection domain of the present invention is not limited to described content.In embodiment, method if no special instructions, carry out routinely, the reagent that uses if no special instructions reagent to be conventional commercial reagent or to prepare according to a conventional method by operation.
Embodiment 1: the MgCl that promotes India canna nitrate nitrogen absorption and assimilation
2the application of promoter, comprises the steps:
(1) experiment material is India canna seedling; Select the India canna seedling of plant height 30-40cm in wetland soil to dig out, be placed on water planting in running water, change every three days water one time, cultivate about 30 days extremely new roots and grow up to for this experiment;
(2) preparation amount of substance concentration is the MgCl of 150 μ mol/L
2solution;
(3) India canna in (1) is placed in to above-mentioned steps (2) MgCl
2in solution, India Canna Rhizome is carried out to 8h pretreatment; Then the plant of processing is transferred to volume and be in the agricultural wastewater that 3L contains different nitrates; Take without the pretreated India canna of superchlorination magnesium is contrast (CK) simultaneously, is also placed in volume and is in the agricultural wastewater that 3L contains different nitrates; Measure the impact of magnesium chloride on nitrate nitrogen efficiency and relative physiologic index in India canna absorption agricultural wastewater.
Embodiment 2: the India canna checking MgCl after adopting embodiment 1 to process
2promoter absorbs the impact of nitrate speed on India canna, comprise the steps:
At tri-time points of 2h, 12h, 22h of processing, get respectively agricultural wastewater water sample 5 mL of experimental group and the processing of control group India canna, add 2 aluminium hydroxide suspension, standing flocculation, centrifugal rear supernatant is measured for nitrate, it during mensuration, with optical length, is the quartz colorimetric utensil of 10 mm, take fresh deionized water as reference, at 220 nm and 275 nm wavelength places, detect respectively, the corrected value of nitrate nitrogen absorbance is that OD220 value deducts the OD275 value of 2 times, checks in nitrate from calibration curve.3 repetitions are done in each processing, get the mean value of measurement result; By the method, verify MgCl
2activator is used the impact that India canna is absorbed to nitrate speed.
Fig. 1 demonstration, A figure is without MgCl
2process the absorption result of India Canna Rhizome to nitrate, B figure is through MgCl
2the absorption result of India Canna Rhizome to nitrate after processing.MgCl
2the result that affects that processing absorbs nitrate speed to India canna shows, adopts MgCl of the present invention
2inorganic agent is processed after India canna, and India canna seedling is significantly higher than and does not pass through MgCl the absorption rate of nitrate
2the control group of processing.
Embodiment 3: the India canna checking MgCl after adopting embodiment 1 to process
2promoter is to India canna plasma membrane H
+the impact of-ATPase activity, comprises the steps:
After A, the wastewater treatment of variable concentrations nitrate nitrogen, the root, 0.5 g of leaf texture that get India canna under different disposal, use liquid nitrogen Rapid-Freezing Method, adds Extraction of plasma membrane liquid (the 0.25 mol/L sorbierite of 1 mL after liquid nitrogen grinding is powder, 1 mmol/L EDTA, 5 mmol/L MgSO
4, 10 mmol/L Tris-HCl pH7.4); Homogenate is with 9000 g, and 4 ℃ of centrifugal 20min remove precipitation, and supernatant is through 30000g, 4 ℃ of centrifugal 1h; Collecting precipitation is also suspended in 0.5mL said extracted liquid it, and gained is cytoplasma membrane albumen;
B, by Brodford method, measure plasmalemma protein concentration: in the distilled water of 800 μ L, add the plasmalemma protein of 5 μ L to mix, then the commercial Brodford solution that adds 200 μ L, under OD595 wavelength, detect protein concentration, and calculate volume corresponding to 500 μ g plasmalemma protein;
C, H
+in the reaction system that is determined at 0.5 mL of-ATPase activity, carry out; Reaction system comprises 50 mmol/L BTP/MES, 5 mmol/L MgSO
4, 50 mmol/L KCl, 0.02% Brij-35 (w/v), 50 mmol/L KNO
3, 1 mmol/L(NH
4)
2moO
4, 1 mmol/L NaN
3, 4 mmol/L ATP-Na
2, start reaction after adding the plasmalemma protein extract of 500 μ g;
Reactant mixture is placed in after 30 ℃ of water-bath 30min, adds reaction terminating liquid 1 ml (2 % H
2sO
4(v/v), 5 % SDS (w/v) and 0.7 % (NH
4)
2moO
4(w/v)), add immediately 50 μ L 10% Vc (w/v) and under room temperature, place 20 min, measuring wavelength is the light absorption value at 700 nm places.
D, according to Phos calibration curve, calculate the release content of Phos, the plasma membrane H of 1 unit
+-ATPase activity is defined as: under the reaction condition of 30 ℃, in 1 minute, every milligram of albumen catalysis ATP decomposes the micromole's number that discharges inorganic phosphate.
Fig. 2 demonstration, A figure is MgCl
2plasma membrane H in India Canna Rhizome before and after processing
+-ATPase activity change, wherein without MgCl
2the India canna of processing is control group, represents, through MgCl with CK
2the India canna of processing is experimental group; B figure is MgCl
2plasma membrane H in India canna leaf before and after processing
+-ATPase activity change, wherein without MgCl
2the India canna of processing is control group, represents, through MgCl with CK
2the India canna of processing is experimental group.MgCl
2processing is to India canna plasma membrane H
+the result that affects of-ATPase activity shows, adopts MgCl of the present invention
2inorganic agent is processed after India canna, plasma membrane H in India canna shoot root and leaf
+-ATPase activity is significantly higher than control group CK.
Embodiment 4: the India canna checking MgCl after adopting embodiment 1 to process
2promoter is to H in India Canna Rhizome
+the impact of-pump activity, comprises the steps:
After A, the wastewater treatment of variable concentrations nitrate nitrogen, get root 0.5 g of India canna under different disposal, use liquid nitrogen Rapid-Freezing Method, after liquid nitrogen grinding is powder, add Extraction of plasma membrane liquid (0.25 mol/L sorbierite, 1 mmol/L EDTA, the 5 mmol/L MgSO of 1 mL
4, 10 mmol/L Tris-HCl pH7.4); Homogenate is with 9000g, and 4 ℃ of centrifugal 20 min remove precipitation, and supernatant is through 30000g, 4 ℃ of centrifugal 1h; Collecting precipitation is also suspended in 0.5ml said extracted liquid it, and gained is cytoplasma membrane albumen;
B, use Brodford method are measured plasmalemma protein concentration.In the distilled water of 800 μ L, add the plasmalemma protein extract of 5 μ L to mix, then add the commercial Brodford solution of 200 μ L, under OD595 wavelength, detect protein concentration, and calculate volume corresponding to 100 μ g plasmalemma protein;
In C, reaction system 1.5 ml, contain 5 mmol/L BTP/MES (pH 6.0), 12 μ mol/L AO, 300 mmol/L KCl, 250 mmol/L sucrose, 0.5 mmol/L EGTA(is used BTP to adjust pH to 6.0), 1 mmol/L NaN
3, 1 mmol/L Na
2moO
4, 50 mmol/L KNO
3, 0.05% Brij-35 (w/v) and 100 μ g plasmalemma proteins; Add abstergent Brij-35 and make the upset of original position film, reaction mixture is at room temperature placed after 20 min, adds 5 mmol/L ATP/BTP (pH 6.0) to start reaction;
D, proton are to carry out in the method for 492 nm light absorption value cancellation according to measuring acridine orange (AO) from outside pumping in film; By the method, verify MgCl
2activator is used H in India Canna Rhizome
+the impact of-pump activity.
Fig. 3 demonstration, A figure is not for passing through MgCl
2the control group of processing, B figure is through MgCl
2the experimental group of processing.Through MgCl
2processing is to India canna root H
+the result that affects of-pump activity shows, adopts MgCl of the present invention
2inorganic agent is processed after India canna, India canna root H
+-pump activity is significantly higher than control group CK.
Embodiment 5: the India canna checking MgCl after adopting embodiment 1 to process
2the impact of promoter on India canna nitrate reductase activity, comprises the steps:
After A, the wastewater treatment of variable concentrations nitrate nitrogen, get root, the 0.5g of leaf texture of India canna under different disposal, use liquid nitrogen Rapid-Freezing Method, after liquid nitrogen grinding is powder, add 1.5mL extract (25 mmol/L phosphate buffer pH7.4,1 mmol/L EDTA, 10 mmol/L cysteines) grind homogenate, be transferred in 2 mL EP pipes 4000 rpm, 4 ℃, centrifugal 15 min, remove precipitation supernatant and are zyme extract;
B, get 0.2 mL zyme extract in 2 mL EP pipes, add 0.5 mL 0.1 mmol/L KNO
3phosphate buffer and 0.3 mL NADH solution, mix, 25 ℃ of water-bath 30 min, after finishing, water-bath adds 0.5 mL sulfanilamide (SN) cessation reaction, add again 0.5 mL alpha-naphthylamine, mix, after colour developing 15 min, with spectrophotometer, in 520 nm places, carry out colorimetric estimation, write down OD value, using and do not add NADH(and add 0.2mL water) as blank;
C, according to calibration curve, calculate reactant liquor Central Asia nitrate nitrogen content, the nitrite nitrogen micrograms producing with every gram of fresh weight per hour represents nitrate reductase activity;
Fig. 4 demonstration, A figure is MgCl
2before and after processing, in India Canna Rhizome, nitrate reductase activity changes, wherein without MgCl
2the India canna of processing is control group, represents, through MgCl with CK
2the India canna of processing is experimental group; B figure is MgCl
2before and after processing, in India canna leaf, nitrate reductase activity changes, wherein without MgCl
2the India canna of processing is control group, represents, through MgCl with CK
2the India canna of processing is experimental group.MgCl
2processing shows the result that affects of India canna nitrate reductase activity, adopts MgCl of the present invention
2inorganic agent is processed after India canna, and in India Canna Rhizome, leaf, the activity of nitrate reductase is significantly higher than control group CK.
Embodiment 6: the India canna checking MgCl after adopting embodiment 1 to process
2the impact of promoter on India canna activity of glutamine synthetase, comprises the steps:
After A, the wastewater treatment of variable concentrations nitrate nitrogen, the root, the 0.5g of leaf texture that get India canna under different disposal, use liquid nitrogen Rapid-Freezing Method, adds 1.5 mL extracts to grind homogenate after liquid nitrogen grinding is powder, transfer them to 4500 rpm/min in centrifuge tube, 4 ℃ of centrifugal 10 min, abandon precipitation, get supernatant centrifugal 10 min of 4500 rpm again, be precipitated as chloroplast part, get centrifugal 20 min of supernatant 12000rpm at 4 ℃, be precipitated as mitochondria part, supernatant is crude enzyme liquid;
B, by Brodford method, measure thick liquid protein concentration, in the distilled water of 800 μ L, add the GS albumen crude enzyme liquid of 5 μ L to mix, then the commercial Brodford solution that adds 200 μ L detects protein concentration under OD595 wavelength, and calculates 0.7 mL crude enzyme liquid soluble protein content;
C, get the thick zyme extract of 0.7 mL and 0.7 mL ATP (40 mmol/L) solution adds (80 mmol/L hydroxylamine hydrochlorides, 80 mmol/L MgSO in 1.6 mL reactant liquor B
4, 20 mmol/L sodium glutamates, 20 mmol/L cysteines, 2 mmol/L EGTA, 0.1 mol/L Tris-HCl pH7.4), put upside down and mix, 37 ℃ of standing 30 min, add 1ml developer (0.2 mol/L TCA, 0.37 mol/L FeCl
3with 0.6 mol/L HCl mixed liquor), put upside down and mix, centrifugal 10 min of 5000 rpm, get supernatant and read absorbance at 540nm place.With not overstriking zyme extract, add 0.7mL reactant liquor A(80 mmol/L MgSO
4, 20 mmol/L sodium glutamates, 20 mmol/L cysteines, 2 mmol/L EGTA, 0.1 mol/L Tris-HCl pH7.4) and as blank;
D, GS vigor (A/mg proteinh)=OD540nm/(soluble protein content * 0.5) calculate enzyme and live; By the method, verify MgCl
2promoter is respectively organized the impact of activity of glutamine synthetase on India canna.
Fig. 5 demonstration, A figure is MgCl
2before and after processing, in India Canna Rhizome, activity of glutamine synthetase changes, wherein without MgCl
2the India canna of processing is control group, represents, through MgCl with CK
2the India canna of processing is experimental group; B figure is MgCl
2before and after processing, in India canna leaf, activity of glutamine synthetase changes, wherein without MgCl
2the India canna of processing is control group, represents, through MgCl with CK
2the India canna of processing is experimental group.MgCl
2processing shows the result that affects of India canna activity of glutamine synthetase, adopts MgCl of the present invention
2inorganic agent is processed after India canna, and in India Canna Rhizome and leaf, the activity of glutamine synthelase is significantly higher than control group CK.
Embodiment 7: the MgCl that promotes India canna nitrate nitrogen absorption and assimilation
2the application of promoter, comprises the steps:
(1) preparation amount of substance concentration is the MgCl of 130 μ mol/L
2solution;
(2) select the India canna seedling of plant height 30-40cm in wetland soil to dig out, be placed in running water and cultivate, change every three days water one time, cultivate about 30 days extremely new roots and grow up to for this experiment;
(3) India canna in (2) is placed in to above-mentioned steps (1) MgCl
2in solution, India Canna Rhizome is carried out to 6h pretreatment; Then the plant of processing is transferred to volume and be in the agricultural wastewater that 3L contains different nitrates; Take without the pretreated India canna of superchlorination magnesium is contrast (CK) simultaneously, is also placed in volume and is in the agricultural wastewater that 3L contains different nitrates; Measure MgCl
2promoter absorbs nitrate speed to India canna, and method is with embodiment 2.
Fig. 6 demonstration, A figure is not for passing through MgCl
2the control group of processing, B figure is through MgCl
2the experimental group of processing.MgCl
2the result that affects that processing absorbs nitrate speed to India canna shows, adopts MgCl of the present invention
2inorganic agent is processed after India canna, India canna seedling to the absorption rate of nitrate higher than control group CK.
Embodiment 8: the MgCl that promotes India canna nitrate nitrogen absorption and assimilation
2the application of promoter, comprises the steps:
(1) preparation amount of substance concentration is the MgCl of 180 μ mol/L
2solution;
(2) select the India canna seedling of plant height 30-40cm in wetland soil to dig out, be placed in running water and cultivate, change every three days water one time, cultivate about 30 days extremely new roots and grow up to for this experiment;
(3) India canna in (2) is placed in to above-mentioned steps (1) MgCl
2in solution, India Canna Rhizome is carried out to 7h pretreatment; Then the plant of processing is transferred to volume and be in the agricultural wastewater that 3L contains different nitrates; Take without the pretreated India canna of superchlorination magnesium is contrast (CK) simultaneously, is also placed in volume and is in the agricultural wastewater that 3L contains different nitrates; Measure MgCl
2promoter absorbs nitrate speed to India canna, and method is with embodiment 2.
Fig. 7 demonstration, A figure is not for passing through MgCl
2the control group of processing, B figure is through MgCl
2the experimental group of processing.MgCl
2the result that affects that processing absorbs nitrate speed to India canna shows, adopts MgCl of the present invention
2inorganic agent is processed after India canna, India canna seedling to the absorption rate of nitrate higher than control group CK.
Claims (2)
1.MgCl
2application as plant nitrate nitrogen sorbefacient.
2. MgCl according to claim 1
2application as plant nitrate nitrogen sorbefacient, is characterized in that: adopting amount of substance concentration is the MgCl of 130-180 μ mol/L
2solution is processed 6-8h to plant, subsequently plant is placed in the agricultural wastewater containing nitrate nitrogen to the growth indexes of nitrate and plant in detection waste water.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104886128A (en) * | 2015-05-14 | 2015-09-09 | 昆明理工大学 | Application of magnesium ion to improvement of plant photosynthesis efficiency |
| CN112575005A (en) * | 2021-01-04 | 2021-03-30 | 昆明理工大学 | Method for improving heavy metal cadmium stress resistance of tobacco and reducing cadmium enrichment |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112575005A (en) * | 2021-01-04 | 2021-03-30 | 昆明理工大学 | Method for improving heavy metal cadmium stress resistance of tobacco and reducing cadmium enrichment |
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