CN105123019B - Ammonium nitrogen enhancing plant removes the application in liquid formaldehyde pollution - Google Patents
Ammonium nitrogen enhancing plant removes the application in liquid formaldehyde pollution Download PDFInfo
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- CN105123019B CN105123019B CN201510465205.1A CN201510465205A CN105123019B CN 105123019 B CN105123019 B CN 105123019B CN 201510465205 A CN201510465205 A CN 201510465205A CN 105123019 B CN105123019 B CN 105123019B
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- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 title claims abstract description 189
- 239000007788 liquid Substances 0.000 title claims abstract description 28
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 title claims abstract description 17
- 230000002708 enhancing effect Effects 0.000 title claims abstract description 9
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- 235000010469 Glycine max Nutrition 0.000 claims abstract description 34
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 abstract description 15
- 238000010521 absorption reaction Methods 0.000 abstract description 13
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 abstract description 8
- 238000002203 pretreatment Methods 0.000 abstract description 8
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- 238000012545 processing Methods 0.000 description 17
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- Cultivation Of Plants (AREA)
- Treating Waste Gases (AREA)
Abstract
The invention discloses the new application of ammonium nitrogen, specially application of the ammonium nitrogen in enhancing plant removes liquid formaldehyde pollution, in use, using 10mmol/L NH4Cl solution pretreatment black soybean root（Whole strain fresh weight 8g）12h, then with the replacement of 4mmol/L formalins handle, in 24h the absorption of plant PARA FORMALDEHYDE PRILLS(91,95) with without NH4The plant of Cl pretreatments utilizes simultaneously compared to adding 5%13NH is passed through in C nuclear-magnetism scientific discoveries4After Cl pretreatments, plant is to H13CHO metabolism enhancing, ammonium nitrogen is the accelerator of more satisfactory enhancing plant absorption liquid formaldehyde, and the pretreatment of ammonium nitrogen can significantly improve absorptivity of the plant root to liquid formaldehyde, prevent and treat significant to industrial wastewater formaldehyde pollution.
Description
Technical field
The invention belongs to the field of plant purification water pollution, and in particular to improve plant purification liquid formaldehyde to ammonium nitrogen
New application in pollution.
Background technology
Formaldehyde is widely used in the life of resin, plastics, leather, paper and fiber etc. as a kind of basic industrial chemicals
In production, all contain formaldehyde in these device institute effluents.Formaldehyde has active chemical property and biological property, energy and albumen
Matter, nucleic acid and lipid produce nonspecific reaction, it is lost biological function, therefore formaldehyde toxicity is extremely strong.Low dose of Long Term Contact
Amount formaldehyde can cause chronic respiratory disease, cause nasopharyngeal carcinoma, colon cancer, the cancer of the brain, paramenia, pregnancy syndrome, neonate
The gene mutation of chromosome abnormality, nucleus, DNA single-stranded interior commissure and DNA are remembered with protein commissure, leukaemia, teenager
Power and intelligence decline etc..
Because the harm of formaldehyde pollution is very extensive, thus have many researchs be directed to exploitation and control formaldehyde pollution technology and
Method, in numerous administering methods, the bioreactor based on microorganism and plant absorption formaldehyde capability development be a kind of row it
The method of efficient and cost-effective material benefit, this kind of reactor can be for the liquid of purifying contaminated in the environment, solid-state and gaseous formaldehyde.
In the system of potted plant purifying formaldehyde, formaldehyde on the one hand can be by the supracutaneous stomata of hole skin and blade on stem by sky
The formaldehyde absorption polluted in gas enters in plant, on the other hand can also be absorbed and be polluteed in soil by under ground portion root
Formaldehyde.Kim etc. (2008) research show it is potted plant it is anistree (Fatsia japonica) and banyan (Ficus benjamina)
Plant daytime root remove formaldehyde amount it is identical with aerial part, but it is 10 times of aerial part that evening root, which removes the amount of formaldehyde,
Illustrate that the removal of root system of plant PARA FORMALDEHYDE PRILLS(91,95) has important contribution.Many results of study show that different plant varieties have different formaldehyde
Absorbability.In order to better profit from plant purification formaldehyde pollution, there are many study tours plant different tissues and organ to inhale
Receive the dynamics of formaldehyde.Blade PARA FORMALDEHYDE PRILLS(91,95) when Zeng etc. (2013) research shows to handle in vitro Leaf of banana with liquid formaldehyde
Uptake and the relation of processing time are square root function, Song etc.（2010）Liquid is absorbed to model plant tobacco and arabidopsis
The dynamic analysis result of formaldehyde illustrates that arabidopsis is better than tobacco to liquid formaldehyde absorbability, and Arabidopsis plant is at formaldehyde
The amount and the relation of processing time for managing absorption formaldehyde in liquid are power function.Xu etc. (2010) etc. is investigated with macro-kinetic model
The dynamic process of Degradation Formaldehyde, passes through kinetic simulation in the biological filter system that the matrix such as plant, microorganism and haydite are constituted
Type predicts that the optimum condition of filtration system removing formaldehyde is consistent with experiment value.
One of a great number of elements necessary to nitrogen is plant, the nitrogen that usual plant absorbs in growth and development process is higher than other ore deposits
Prime element, thus influence of the nitrogen to growth and development of plants is larger.Ammonium nitrogen is a kind of available host inorganic nitrogen shape of plant
State.There are some researches show in Leaf Vegetables in Liquid Culture suitably result in more preferable yield with applying ammonium nitrogen.Gao Zhi is built（2015）Hair
Now leaf stomatal conductance, reduction Transpiration, increase leaf water can be increased using ammonium nitrogen to utilize.Conventional is a lot
Result of study confirms that the related enzyme of overexpression formaldehyde metabolism can improve plant generation in genetically modified plants by genetic manipulation
Thank, absorb and purifying formaldehyde ability.But change the formaldehyde metabolism and the research of absorbability of plant with Physiology and biochemistry method
It yet there are no relevant report.Therefore whether the application of this study tour ammonium nitrogen can promote the absorption of black soybean root system PARA FORMALDEHYDE PRILLS(91,95),
Excavating a kind of reinforcing agent that can improve black soybean root purification liquid formaldehyde pollution ability has important environment protection significance.
The content of the invention
Strengthening black big it is an object of the invention to provide a kind of reinforcing agent that liquid formaldehyde purification absorbs, i.e. ammonium nitrogen of improving
Beans（Glycine max var.）Purify the effect in liquid formaldehyde pollution.
In order to realize the above-mentioned purpose of the present invention, technical scheme is as follows：
（1）Select the autumn sowing black soybean full seed in Yunnan to rinse epidermis with deionized water at normal temperature, be laid in afterwards
In the culture dish for being lined with moistening filter paper, the dark Germination under 25 DEG C of constant temperature treats that root grows about 3cm and is transferred to pin hole eye
Thin cystosepiment on, be placed in the black plastic basin for filling Hoagland ' s nutrient solutions, in 25 DEG C of temperature, be with intensity daily
1200μmol·m-2·s-1（Intensity of illumination) the h of strong illumination 12 carry out floating culture, every 1 day change a nutrient solution；
（2）The consistent plant of growing way is selected to weigh whole strain fresh weight common 8g when black soybean grows to two pairs of blades（4 plant are fresh
Weight）, with the NH of 75ml various concentrations4Cl solution（3、6、10mmol/L）Pre-process after 12h, then with 75ml 4mmol/L formaldehyde
Solution is replaced and continued with, and determines different time（4、12、24h）Residual formaldehyde content in solution.Each processing sets five weights
It is multiple；NH is selected according to measurement result4The optium concentration of Cl pretreatments.
（3）Use step（2）In the black soybean of identical growing state weigh the whole common 8g of strain fresh weight, use 10mmol/L NH4Cl is molten
The ml of liquid 75 carries out pretreatment 12h to black soybean root, then uses 4mmol/L H13CHO solution replaces processing 2h, takes root 2g profits
With13C nuclear-magnetisms determine metabolite；Control CK is used as with the black soybean root 2g without any processing；Use 4mmol/L H13CHO is molten
Liquid processing 2h root is compareed as another.
The ammonium nitrogen that the present invention is provided as black soybean root absorption liquid formaldehyde accelerator, it is easy to use, into
This is cheap.Ammonium nitrogen significantly improves the ability of plant purification formaldehyde in waste water pollution, opens and promotes to plant using ammonium nitrogen
Thing administers the new way of formaldehyde pollution, contributes to scientific worker to promote plant purification formaldehyde in waste water pollution capacity to ammonium nitrogen
Molecular mechanism research, have broad prospects, also open in plant purification industrial production wastewater formaldehyde pollution prevention and control field
Industrial crops play the blank of new role in pollution field of purification.
Beneficial effects of the present invention：Ammonium nitrogen enhancing plant root of the present invention absorbs liquid formaldehyde, with input
It is low, simple to operate, the characteristics of purification efficiency is high；Ammonium nitrogen is the accelerator of more satisfactory enhancing plant absorption liquid formaldehyde,
The pretreatment of ammonium nitrogen can significantly improve absorptivity of the plant root to liquid formaldehyde, to the anti-tool of industrial wastewater formaldehyde pollution
It is significant.
Brief description of the drawings
Fig. 1 is various concentrations NH4Cl（3、6、10mmol/L）Influence to black soybean root liquid formaldehyde absorption dynamics；
Wherein A figures are residual formaldehyde content of the formaldehyde processing liquid in 4,12,24h；B figures are first of the black soybean root in 4,12,24h
Aldehyde uptake；Wherein volatilization HCHO represents the content of formaldehyde of volatilization；Absorption HCHO represents the suction of fixing black soybean root PARA FORMALDEHYDE PRILLS(91,95)
It is attached；Control represents the control without any processing；NH4 +- 3mM represents 3mmol/L NH4Cl, NH4 +- 6mM represents 6mmol/L
NH4Cl, NH4 +- 10mM represents 10mmol/L NH4Cl；
Fig. 2 is13C nmr analysis NH4Cl pretreatments are exposed to 4mmol/L H to black soybean root13Shadow is metabolized in CHO solution
Loud full spectrum；Wherein Ref is maleic acid internal reference signal peak, is compareed as the black soybean root without any processing；H13CHO 2h tables
Show by 4mmol/L H13CHO handles 2h black soybean root；NH4Cl pretreatment 12h+H13CHO 2h represent black soybean root
First pass through 10mmol/L NH4Cl pre-processes 12h, then using 4mmol/L H13CHO solution replaces processing 2h；
Fig. 3 is in the present invention13C nmr analysis NH4Cl pretreatments are exposed to 4mmol/L H to black soybean root13CHO solution
That listed metabolic intermediate is Icit respectively in the influence of middle metabolic intermediate, figure（Isocitric acid）, Cit（Lemon
Acid）, FA（Formic acid）, Ser（Serine）, Gly（Glycine）；Wherein internal reference Ref（Horse Lay acid）Signal peak is 130.66 ppm；
Fig. 4 integrates for Icit signal peaks in the present invention, and its signal peak corresponds to Icit5,182.01 ppm；Icit6,
181.39 ppm；Icit1,178.96 ppm；
Fig. 5 integrates for Cit signal peaks in the present invention, and its signal peak corresponds to Cit6,179.95 ppm；Cit3,70.11
ppm；Cit (2,4), 42.65 ppm；
Fig. 6 integrates for FA signal peaks in the present invention, and its signal peak corresponds to FA, 171.20 ppm；
Fig. 7 integrates for Ser signal peaks in the present invention, and its signal peak corresponds to [3-13C] Ser, 61.10 ppm；[2-13C]
Ser, 55.26 ppm；
Fig. 8 integrates for Gly signal peaks in the present invention, and its signal peak corresponds to [2-13C] Gly, 40.23 ppm.
Embodiment
The present invention is described in further detail below by embodiment and accompanying drawing, but the scope of the present invention is not limited to
The content.Unless otherwise specified, routinely operation is carried out method, the use of reagent is unless otherwise specified routine in embodiment
Purchase reagent or the reagent prepared according to a conventional method.
Embodiment 1：The water planting of black soybean plant and processing.Step is as follows：
1st, experiment material is black soybean water planting seedling, is seeded in after black soybean vernalization on perforated plastic thin plate, is used
Hoagland ' s suspension cultures, change a nutrient solution every other day, and this experiment is used for when seedling grows two pairs of blades；
2nd, various concentrations are configured（3、6、10mmol/L）NH4The HCHO solution and H of Cl solution and 4mmol/L13CHO is molten
Liquid；
3rd, with 3,6,10mmol/L NH4Cl solution 75ml respective pretreatment black soybean root（The whole common 8g of strain fresh weight）12h；
Treat NH4After Cl pretreatment times terminate, NH is outwelled4Cl treatment fluids, are separately added into 75ml 4mmol/L formalin（It is denoted as just
Beginning HCHO, base level is 100%）Continue with, determine different time（4、12、24h）Residual formaldehyde content in solution（Determine
Residual formaldehyde content is denoted as remaining HCHO%）, it is denoted as while not being put into plant under identical processing mode and determining formaldehyde volatile quantity
Volatilize HCHO%, and the black soybean root measure residual formaldehyde content that fixing is put under identical processing mode is denoted as adsorbing HCHO%
（Fixing refers to Fresh Plants being placed in 105 DEG C of high temperature lower 30 minutes, the enzyme in plant is lost activity, prevent metabolism from continuing
Carry out）, and the base level of formaldehyde processing liquid is denoted as 100%, then the formaldehyde amount of black soybean root absorption is absorptivity %=100%
（Base level）- residue HCHO%-volatilization HCHO%-absorption HCHO%；Blake bottle is using fresh-keeping during experiment process
Film carries out sealing treatment；Experiment process condition is 25 DEG C of continuous illumination intensity（1200 µmol m–2 s–1）；Every group of experiment
It is respectively provided with five independent repetitions；
4th, 10mmol/L NH are used4Cl solution pretreatment black soybean root（The whole common 8g of strain fresh weight）12h, treats NH4Cl is pre-processed
After end, by NH4Cl solution is outwelled, and adds 75ml 4mmol/L H13CHO solution continues with 2h, then going out using flowing
Bacterium distilled water flushing root, blotting paper blots root surface moisture, liquid nitrogen cryopreservation；Take 4mmol/L H13The black soybean of CHO processing
Root 2g takes the black soybean root 2g without any processing as the two of control as one of control；Experiment condition is with step 3.
Experimental example 2：Using the content of residual formaldehyde in the step 3 of Nash method determination experiments example 1；Specific determination step is as follows：
1st, Nash preparation of reagents：About 800 ml distilled water are injected into 1000 ml flasks, 150g ammonium acetates are dissolved, then
3 ml glacial acetic acids and 2 ml acetylacetone,2,4-pentanediones are added, plus distilled water is settled to 1000 ml and fully mixed；By the Nash prepared
Agent transfer to brown bottle is kept in dark place standby；Solution colour can be dimmed in the 12h initially prepared for Nash reagents, therefore fresh
The Nash reagents of preparation are reused after should placing 12h；
2nd, 1 ml reaction system includes Nash reagents 500 μ l, the μ l of distilled water 450 and the μ l of formalin 50, reaction
System is placed in 30 DEG C of min of water-bath 30 after mixing；Using 500 μ l Nash reagents plus 500 μ l distilled water reaction system as
Control zeroing, determines formaldehyde with Nash reagents in OD410Under chromogenic reaction value；
It will be seen from figure 1 that by 10mmol/L NH4After Cl pretreatments 12h, compared to 3,6mmol/L NH4Cl and
Without NH4The control of Cl pretreatments, black soybean root absorption formaldehyde ability enhancing is throughout managed residual formaldehyde in time point solution and contained
Amount reduces obvious（Fig. 1 A）, compared with the control, in 24h, the absorbability of black soybean PARA FORMALDEHYDE PRILLS(91,95) adds 5%（Fig. 1 B）.
Experimental example 3：Using13Metabolite in C-NMR technical Analysis step 4 plant roots, concrete operation step is as follows：
1st, treated black soybean root is rinsed 4-5 times with sterile purified water, then with blotting paper blotting material surface
Residual moisture, liquid nitrogen flash freezer and the grind into powder in mortar add the 100 mM phosphoric acid that 3 mL contain 100 mM horses Lays acid
Potassium buffer solution（KPB, pH 7.4）Extracting；
2nd, extract is centrifuged（12000×g）Remove cell fragment within 10 minutes；
3rd, supernatant freezing is dissolved in the mM kaliumphosphate buffers of 0.5 mL 100 after draining；
4th, centrifuge（12000×g）Take within 3 minutes supernatant to load nuclear magnetic tube, and add 5%2H2O（v/v）, NMR receipts
Collect data.In order to contrast the relative amount of mesostate, according to Ref internal references（It is set as 1）Target peak is integrated.
Fig. 2 is black soybean root13The full spectrum of C nuclear-magnetisms metabolism；Fig. 3 analyzes for metabolic intermediate, it can be seen that pass through NH4Cl
After pretreatment, metabolic intermediate such as isocitric acid in black soybean root（Icit, Fig. 4）, citric acid（Cit, Fig. 5）, formic acid
（FA, Fig. 6）、[2,3-13C]Ser（Fig. 7）[2-13C]Gly（Fig. 8）Dramatically increased compared to control, while as can be seen that except
[2-13C] Gly, by NH4The black soybean root metabolic intermediate relative amount of Cl pretreatments is above only H13CHO processing
Experimental group；This shows that ammonium nitrogen enhances metabolic adsorption of the black soybean root to liquid formaldehyde.
Claims (1)
1. application of the ammonium nitrogen in enhancing plant black soybean removes liquid formaldehyde pollution.
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