CN113184998B - Phenol root secretion for promoting co-metabolism degradation of alkylphenol - Google Patents
Phenol root secretion for promoting co-metabolism degradation of alkylphenol Download PDFInfo
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- CN113184998B CN113184998B CN202110503820.2A CN202110503820A CN113184998B CN 113184998 B CN113184998 B CN 113184998B CN 202110503820 A CN202110503820 A CN 202110503820A CN 113184998 B CN113184998 B CN 113184998B
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/34—Organic compounds containing oxygen
- C02F2101/345—Phenols
Abstract
The method selects root exudates as a co-metabolism substrate of alkylphenol, wherein p-coumaric acid (phenols) and oxalic acid (small molecular organic acids) are respectively used as specific root exudates and non-specific root exudates, common wetland plant reed is used as a tested plant, and the co-metabolism degradation behavior of alkylphenol under the synergistic action of plant-microorganism is discussed by combining a removal effect pilot test and a rhizosphere effect pilot test. In the removal effect experiment, pilot-plant constructed wetland experiments are utilized to illustrate the removal rule and the influence factors of the alkylphenol under different treatment conditions, including the concentration ratio of the root exudates to the alkylphenol, the existence of the root exudates, the types of the root exudates, the existence of plants and the like. In rhizosphere effect experiments, degradation pathways of alkylphenol and microbial response mechanisms thereof are revealed through adsorption degradation experiments, plant absorption experiments, microbial abundance and diversity analysis and enzyme activity analysis.
Description
Technical Field
The invention relates to the technical field of phenol root exudates, in particular to phenol root exudates for promoting co-metabolism degradation of alkylphenol.
Background
As a common chemical raw material, p-tert butyl phenol (PTBP) is widely used for producing phenolic resin and a surfactant, waste water containing the PTBP is collected to a sewage treatment plant through an urban water pipe network system and is discharged into environmental water and soil along with tail water and sludge of the sewage treatment plant, the PTBP is a common environmental endocrine disruptor and has physiological toxicity and estrogen effect, and the wide use of the PTBP causes the substance to be frequently detected in the environment and has adverse effects on the ecological environment, food safety and human health.
Although the PTBP can be effectively degraded by physicochemical methods such as advanced oxidation and the like, the PTBP has the disadvantages of high cost, high energy consumption, easy secondary pollution and the like. The biological method can effectively solve the problems, so the biological method is widely used for urban sewage treatment and environmental water purification, but the biological method still has the defect of long reaction period, particularly for refractory organic pollutants (such as PTBP). Therefore, in order to realize the economic and efficient removal of PTBP, the development of a means capable of effectively promoting the biodegradation of PTBP is urgently needed.
Phenolic Root Secretions (PREs) released by plants have the potential to promote the degradation of aromatic compounds, but the reaction processes and mechanisms involved behind them are not clear. Therefore, representative wetland plants PREs such as p-hydroxybenzoic acid (PHA), p-coumaric acid (PCA), Caffeic Acid (CA), Ferulic Acid (FA) and the like are selected as research objects in the research, and the research aims to verify the promotion effect of the PREs on the degradation of PTBP in the rhizosphere of reed; and the mechanism of action of PREs in influencing PTBP degradation is revealed.
Disclosure of Invention
In order to achieve the purpose, the invention provides the following technical scheme:
the phenol root exudates promote the co-metabolism degradation of the alkylphenol, and the phenol root exudates promote the co-metabolism degradation of the alkylphenol comprises the following steps:
s1: experimental setup and culture solution: eight small-test artificial wetlands are arranged, and each artificial wetland device consists of plastic cylinders with the height of 60cm multiplied by the diameter of 20 cm.
S2: pilot test for removal effect: in order to examine the influence of different influencing factors on the removal of p-tert-butylphenol (PTBP), four groups of treatments were set in total in the experiment, two treatments were set in parallel for each treatment, wherein the influence of different analyte Secretions (REs) on the removal of PTBP was compared by setting the PCA group to which p-coumaric acid was added and the OA group to which oxalic acid was added, the influence of the presence or absence of exogenous root secretions on the removal of PTBP was compared by setting the CK group to which no root secretions were added, and the influence of the presence or absence of plants on the removal of PTBP was compared by setting the UP group to which p-coumaric acid was added and no plants were added.
S3: removal experiments: the device starts to test after the plant growth is stable after the device is operated for half a year in a test mode, the device adopts a discontinuous circulating water inlet mode, and each circulation period is two days.
S4: rhizosphere effect bench test: after the experiment is finished, all the constructed wetland devices are immediately dismantled, plants and soil are taken out, rhizosphere layer soil samples with plant systems and upper layer soil samples without plant systems are respectively collected, then the plants are cleaned by tap water, cleaned by ultrapure water and wiped by a towel for later use, and plant root system tissue samples are collected.
S5: soil adsorption degradation experiment: adding rhizosphere soil sample from a plant system and upper soil sample from a plant-free system into a triangular flask filled with 10mg/LPTBP microorganism inorganic culture solution, plugging a bottle mouth with cotton, placing in an oscillator for two days for culture, setting a sterilization control group (treated by a steam sterilizer before the experiment starts), and measuring the content of PTBP and TOC in culture solution of an unsterilized group and a sterilized group in different time periods.
S6: plant uptake experiments: rinsing a root tissue sample of a reed plant to be tested with a sodium hypochlorite solution for 5min, then washing with sterile deionized water twice to achieve a sterilization effect, autoclaving a plant inorganic culture solution for later use, adding the sterilized plant sample into a triangular flask filled with a 10mg/LPTBP sterilized plant inorganic culture solution, plugging the flask with cotton, then placing the flask in an oscillator for two days, and measuring the content of PTBP and TOC in the culture solution in different time periods.
S7: and (3) analyzing the abundance and diversity of the microorganisms: total DNA of the microorganisms was extracted from each soil sample using a soil genomic DNA extraction kit according to the instructions.
S8: and (3) enzyme activity analysis: the PTBP-related degrading enzyme activities of the rhizosphere soil sample taken from the plant system and the upper soil sample without the plant system, namely, monophenol oxidase, polyphenol oxidase, monooxygenase and dioxygenase, were measured.
S9: statistical analysis: the statistical analysis of this study used SPSS software, where the difference analysis and correlation analysis between two variables were calculated using one-way analysis of variance and pearson correlation coefficients, respectively; wherein, six constructed wetlands are provided with plant groups and planted with reeds, two constructed wetlands are provided with no plant groups, plant seedlings are purchased from local markets and planted in the constructed wetlands in 12 months in 2018 according to the density of 1 plant/bed, and Hoagland nutrient solution is added into the device every week to culture the plant growth before the effect experiment is started.
Preferably, the experimental feed water is 10mg/L of the Hoagland nutrient solution of PTBP. In order to study the influence of concentration ratios of different REs and PTBP on pollutant removal, three REs concentration gradients are set in the experiment, and three RE/PTBP concentration gradients are correspondingly obtained.
Preferably, the operation is repeated for three periods under each RE/PTBP concentration gradient, the operation is performed for one period after the concentration gradient is switched every time, the sampling is started after the device is stabilized, and the water inflow sample with each concentration gradient in 0h and the water outflow samples in 3 h, 12 h, 24 h and 48h are respectively collected.
Preferably, the resulting samples are used in a series of rhizosphere effect bench tests: the soil sample is used for adsorption degradation experiments, microbial abundance and diversity analysis and enzyme activity analysis, the plant root system tissue sample is used for plant absorption experiments, and in addition, the fresh weight and the plant height of the plant are also detected to be used for calculating the plant biomass growth.
Preferably, the sterilized plant sample is added into a triangular flask filled with 10mg/L PTBP sterilized plant inorganic culture solution, the bottle mouth is plugged with cotton, then the triangular flask is placed into an oscillator for culturing for two days, then the culture solution is poured off, the plant sample is taken out and transferred into 100mL of ultrapure water, the flask is sequentially oscillated on an ultrasonic cleaning machine and an eddy oscillator for 1min for six times, and then the content of PTBP and TOC in the obtained suspension is determined and respectively used as the adsorption component of the PTBP root surface and the adhesion component of the TOC root surface.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, the existence of oxalic acid promotes the growth of plants, so that the plant absorption process plays a certain role in removing PTBP, but the existence of coumaric acid inhibits the activity of the plants, so that the plant absorption effect is not greatly influenced in a PCA group, although the addition of coumaric acid prevents the plants from absorbing PTBP, the plant absorption effect can obviously enrich PTBP degradation functional flora, and particularly, the plant absorption effect can catalyze PTBP cleavage reaction by stimulating dioxygenase activity in a plant-free system.
Drawings
Fig. 1 is a design diagram of four groups of artificial wetland treatment devices.
FIG. 2 shows ammonia Nitrogen (NH) in different treatments under different RE/PTBP gradients according to the present invention4-N), Total Organic Carbon (TOC), removal rate constant for p-tert-butylphenol (PTBP).
FIG. 3 is a graph showing the change law of PTBP concentration under unsterilized and sterilized conditions in the soil adsorption degradation experiment of the present invention.
FIG. 4 is a graph showing the concentration change and composition of PTBP in the uptake test of plants according to the present invention.
FIG. 5 is a graph showing the results of analysis of abundance and diversity of the microorganism of the present invention.
FIG. 6 is a graph showing the results of the analysis of the enzyme activity of the present invention (MPO: monophenol oxidase; PPO: polyphenol oxidase; MO: monooxygenase; DO: dioxygenase).
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example one
Referring to fig. 1-4, the present invention provides a phenol root exudate for promoting the co-metabolic degradation of alkylphenol, comprising the following steps:
s1: experimental setup and culture solution: eight small-test artificial wetlands are arranged, and each artificial wetland device consists of plastic cylinders with the height of 60cm multiplied by the diameter of 20 cm.
S2: pilot test for removal effect: in order to investigate the influence of different influence factors on PTBP removal, four groups of treatments are set in the experiment, wherein each treatment is set in two parallel, the influence of different REs on PTBP removal is compared by setting a PCA group added with p-coumaric acid and an OA group added with oxalic acid, and whether the REs have the influence on PTBP removal is verified by setting a blank control CK group (CWS 5-6) without adding any root exudates; the effect of plant presence on PTBP removal was determined by setting the UP group without plant control (CWS 7-8) to which p-coumaric acid was added.
S3: removal experiments: the device starts to test after the plant growth is stable after the device is operated for half a year in a test mode, the device adopts a discontinuous circulating water inlet mode, and each circulation period is two days.
S4: rhizosphere effect bench test: after the experiment is finished, all devices are immediately dismantled, plants and soil are taken out, rhizosphere soil samples with plant systems and upper soil samples without plant systems are respectively collected, then the plants are cleaned by tap water, cleaned by ultrapure water and wiped by a towel for standby, and plant root system tissue samples are collected.
S5: soil adsorption degradation experiment: adding rhizosphere soil samples from a plant system and upper soil samples from a plant-free system into a triangular flask filled with 10mg/L-PTBP microorganism inorganic culture solution, plugging the flask with cotton, placing the flask in an oscillator for two days, setting a sterilization control group (treated by a steam sterilizer before the experiment starts), and measuring the content of PTBP and TOC in culture solutions of an unsterilized group and a sterilized group in different time periods (0, 3, 9, 24 and 48 h).
S6: plant absorption test: the root tissue sample of the reed taken from the tested plant is rinsed for 5min by sodium hypochlorite solution and then rinsed twice by sterile deionized water to achieve the sterilization effect. The plant inorganic culture solution was autoclaved for use, and a sterilized plant sample (10g) was added to a Erlenmeyer flask (500mL) containing 10mg/L of PTBP sterilized plant inorganic culture solution (250mL), and the bottle mouth was closed with cotton, followed by culturing in a shaker (120rpm, 25 ℃ C.) for two days. The content of PTBP and TOC in the culture solution is measured in different time periods (0, 3, 9, 24 and 48 h).
S7: and (3) analyzing the abundance and diversity of the microorganisms: total DNA of the microorganisms was extracted from the soil samples using a soil genomic DNA extraction kit according to the instructions.
S8: and (3) enzyme activity analysis: the PTBP-related degrading enzyme activities of the rhizosphere soil sample taken from the plant system and the upper soil sample without the plant system, namely, monophenol oxidase, polyphenol oxidase, monooxygenase and dioxygenase, were measured.
S9: statistical analysis: statistical analysis of this study used SPSS software. Wherein, the difference analysis and the correlation analysis between the two variables are respectively calculated by using single-factor variance analysis and Pearson correlation coefficient.
In the present invention, the concentration of phenolic compounds is determined using a high performance liquid chromatograph, and the concentration of TOC is determined using a total organic carbon analyzer; microbial cell density was determined using an ultraviolet spectrophotometer.
In the invention, six constructed wetlands are planted with reed and plant group, two constructed wetlands are not planted with reed, plant seedlings are purchased from local markets and planted in the constructed wetlands in 12 months in 2018 according to the density of 1 plant/bed, and before the effect experiment is started, Hoagland nutrient solution is added into the device every week to culture the plant growth.
In the present invention, the experimental feed water was 10mg/L of the Hoagland nutrient solution of PTBP. In order to study the influence of concentration ratios of different REs and PTBP on pollutant removal, three REs concentration gradients are set in the experiment, and three RE/PTBP concentration gradients are correspondingly obtained.
In the invention, the operation is repeated for three periods under each RE/PTBP concentration gradient, the operation is performed for one period after the concentration gradient is switched every time, the sampling is started after the device is stabilized, and the water inlet samples with each concentration gradient of 0h and the water outlet samples with each concentration gradient of 3 h, 12 h, 24 h and 48h are respectively collected.
In the present invention, the resulting samples were used in a series of rhizosphere effect bench tests: the soil sample is used for adsorption degradation experiments, microbial abundance and diversity analysis and enzyme activity analysis, the plant root system tissue sample is used for plant absorption experiments, and in addition, the fresh weight and the plant height of the plant are also detected to be used for calculating the plant biomass growth.
In the invention, a sterilized plant sample is added into a triangular flask filled with 10mg/L PTBP sterilized plant inorganic culture solution, a bottle mouth is plugged by cotton, then the triangular flask is placed in an oscillator for culturing for two days, then the culture solution is poured out, the plant sample is taken out and transferred into 100mL of ultrapure water, the plant sample is sequentially oscillated on an ultrasonic cleaning machine and an eddy oscillator for 1min for six times, and then the content of PTBP and TOC in the obtained suspension is measured and respectively used as a PTBP root surface adsorption component and a TOC root surface adhesion component.
1. As a result: the growth condition of the reed is good in the whole experiment period, the root system is developed, the root zone is expanded to the periphery to occupy the whole rhizosphere layer, the growth amounts of the fresh weight and the plant height of the plant are 489-778 g and 55-87 cm respectively, wherein the biomass growth amount of the OA group is higher than that of the CK group, and the biological growth amount of the PCA group is lower than that of the CK group.
2. Physicochemical parameters: along with the increase of RE/PTBP concentration gradient, the inflow load of acidic REs is increased, the pH of inflow water is sharply reduced from 6.58 to 3.77 in an OA group, and is slightly reduced from 6.67 to 6.29 in a PCA group, but the pH of outflow water is close to neutral after being adjusted by a wetland system, which shows that the wetland system has strong neutralizing capacity for strong-acid oxalic acid or weak-acid p-coumaric acid, and the whole device is in an anoxic environment due to the fact that the wetland system is in a submerged state, so that relatively low ORP is beneficial to the activity of denitrifying bacteria.
3. Hydration parameters: according to fig. 1, NH when RE/PTBP ═ 1:14The removal rates of N, TOC and PTBP in each treatment group are not greatly different and are respectively 0.28-0.69/d, 0.09-0.21/d and 0.81-1.08/d, and when RE/PTBP is 5:1, NH4The removal rate of-N in the plant-containing group is higher than that in the plant-free group, and the removal rate in the-PREs group is higher than that in the + PGroup of REs. The removal rate of TOC in the + PREs group is higher than that of the-PREs group, each reed bed system obtains higher removal rate to PTBP, and NH is added when RE/PTBP is 10:14The removal rate of-N in the plant-containing group is higher than that in the plant-free group, the removal rate of TOC in the + PREs group is higher than that in the-PREs group, the removal rate of PTBP in the plant-containing group is higher than that in the plant-free group, and the removal rate in the-PREs group is higher than that in the + PREs group.
4. The removal effect was compared under different concentration gradients: NH (NH)4N can be effectively removed under each concentration ratio and each treatment condition, the removal rate at the medium-high concentration ratio is higher than that at the low-high concentration ratio, the removal rate at the plant group is higher than that of the plant-free group, TOC can be effectively removed in the + PREs group with medium-high concentration gradient but can not be effectively removed in the-PREs group and the low concentration gradient, which is probably related to the low TOC water inlet load of the-PREs group and the low concentration ratio, PTBP can be effectively removed under each concentration ratio and each treatment condition, and the removal rate at the medium concentration ratio is the highest, which indicates that the growth of degradation functional flora can be stimulated by adding a certain amount of growth substrate, key enzymes beneficial to the co-metabolism of the refractory organic pollutants can be released, however, when the growth substrate is too much, competitive inhibition can be caused, so that the removal rate of the pollutants is slowed down, and the removal reaction rules of each pollutant are very similar, are removed mainly in the first 3 hours of entering the system, after which the removal rate gradually slows down, and the reaction almost stops after 24 hours. Corresponding thereto, NH4The concentration changes among-N, TOC, PTBP were found to be in significant positive correlation, indicating that the abatement reactions between these contaminants cross each other, interact.
5. Rhizosphere effect bench test: 1. according to FIG. 2: wherein, the organic pollutants of the unsterilized group can be removed through two ways of soil adsorption and the degradation of microorganisms on a biological membrane, the organic pollutants of the sterilized group can only be removed through the soil adsorption, the removal rate of PTBP between different treatment systems is not greatly different, but the removal rate of PTBP in the unsterilized group (62-71%) is higher than that of the sterilized group (28-35%), according to a liquid phase map, almost no PTBP miscellaneous peak appears in the unsterilized group, and the PTBP miscellaneous peak frequently appears in the sterilized group during the whole experiment. This shows that the PTBP of the unsterilized group is degraded by the open loop of the microorganism after being adsorbed by soil, while the PTBP of the sterilized group is difficult to break by the open loop due to lack of biodegradation, so that a plurality of benzene derivatives are generated; 2. according to FIG. 3: the PTBP content precipitated from the plant root system after the ultrasonic and oscillation processes is used as a root surface adsorption component, the difference value obtained by subtracting the root surface adsorption component from the total PTBP removal amount is used as a plant adsorption component, the removal rate of the PTBP in a PCA group is only 16%, about 56% of the removed PTBP is a plant adsorption component, and 44% is a root surface adsorption component.
6. And (3) analyzing the abundance and diversity of the microorganisms: according to fig. 4, wherein the abundance of the microorganisms is represented by a Chao 1 index, the larger the index is, the more the species number of the sample is, the higher the abundance is; the diversity of the microorganisms is expressed by using a Shannon index, the larger the index is, the more species types and the higher diversity of the samples are shown, the abundance and the diversity of the bacteria in the + REs group are higher than those in the-REs group, the bacteria growth can be effectively promoted and the bacterial types can be enriched by adding REs by external aid, generally, the bacterial content of a plant-free system is obviously lower than that of a plant-based system, the abundance and diversity of bacteria in the plant-free system added with p-coumaric acid in the research are not obviously different from those in the plant system, therefore, the release of the REs is a key factor for promoting the growth and the activity of rhizosphere bacteria of plants, the REs are added by external source to generate the effect similar to the plant, the abundance and diversity of the fungus in each treatment system have no obvious change, this may suggest that the fungus is relatively stable in reed bed systems and not susceptible to the addition of exogenous REs.
7. And (3) enzyme activity analysis: according to fig. 5, polyphenol oxidase and monophenol oxidase were classified by nature, and monooxygenase and dioxygenase were classified by function, wherein the monophenol oxidase activity was significantly higher in the + PREs group than in the-PREs group, the polyphenol oxidase activity was significantly higher in the + REs group than in the-REs group, the monooxygenase activity did not change much in each treatment group, and the dioxygenase activity was significantly higher in the plant-free system to which PCA was added, and further, the polyphenol oxidase activity and the bacterial abundance were significantly positively correlated in each treatment group, and the monooxygenase activity and the fungal abundance were significantly positively correlated.
8. Discussion: in the research, the removal rate of PTBP when RE/PTBP is 5:1 can reach 88-98% at most, because RE/PTBP gradient can not only influence co-metabolism to improve the removal rate of PTBP, but also can slow down the removal rate of PTBP through competitive inhibition, soil adsorption, microbial degradation and plant absorption are considered as main removal ways of PTBP in a wetland system, although REs are added to have little influence on the community distribution of fungi, the community abundance and diversity of bacteria are greatly changed, thereby the PTBP is promoted to be used as a carbon source by anaerobic ammonium oxidation bacteria and used as an electron donor by denitrifying bacteria, the REs can strengthen the activity of non-specific enzymes, but only coumaric acid can stimulate the release of specific enzymes, further the hydroxylation process of PTBP and the subsequent ring-opening cracking process are promoted, the growth of plants is promoted by the existence of oxalic acid, the plant absorption process also plays a certain role in removing the PTBP, but the plant activity is inhibited by the existence of the coumaric acid, so the plant absorption effect has little influence in a PCA group, although the plant absorption effect is hindered by the addition of the coumaric acid, the plant absorption effect can obviously enrich PTBP degradation functional flora, and particularly the plant-free system can catalyze the PTBP cleavage reaction by stimulating the activity of dioxygenase.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (5)
1. A research method for promoting co-metabolism and degradation of alkylphenol by phenol root exudates is characterized in that: the method comprises the following steps:
s1: experimental setup and culture solution: arranging eight small-test constructed wetlands, wherein each constructed wetland device consists of plastic cylinders with the height of 60cm multiplied by the diameter of 20 cm;
s2: pilot test for removal effect: in order to investigate the influence of different influence factors on the removal of p-tert-butylphenol (PTBP), four groups of treatments are arranged, wherein each treatment is arranged in two parallel, wherein the influence of different root exudates REs on the removal of PTBP is compared by arranging a PCA group added with p-coumaric acid and an OA group added with oxalic acid, the influence of the existence of exogenous root exudates on the removal of PTBP is compared by arranging a CK group not added with any root exudates, and the influence of the existence of plants on the removal of PTBP is compared by arranging an UP group added with p-coumaric acid and no plants; wherein, six constructed wetlands are provided with plant groups, the plant is reed, two constructed wetlands are provided with no plant groups, plant seedlings are purchased from local markets and planted in the constructed wetlands according to the density of 1 plant/bed, and Hoagland nutrient solution is added into the device every week to culture the plant growth before the effect experiment is started;
s3: removal experiments: the device starts the test after the plant growth is stable after the device is operated for half a year in a test mode, the device adopts a discontinuous circulation water inlet mode, and each circulation period is two days;
s4: rhizosphere effect bench test: after the experiment is finished, all the constructed wetland devices are dismantled, plants and soil are taken out, rhizosphere layer soil samples with plant systems and upper layer soil samples without plant systems are respectively collected, then the plants are cleaned by tap water, cleaned by ultrapure water and wiped by a towel for later use, and plant root system tissue samples are collected;
s5: soil adsorption degradation experiment: respectively adding rhizosphere soil samples of a self-contained plant system and upper soil samples of a non-contained plant system into a triangular flask filled with a microorganism inorganic culture solution, wherein the microorganism inorganic culture solution contains 10mg/LPTBP, plugging a bottle opening with cotton, then placing the bottle opening into an oscillator for two days, setting a sterilization control group at the same time, and measuring the contents of PTBP and TOC in culture solutions of a non-sterilized group and a sterilized group in different time periods;
s6: plant uptake experiments: rinsing a root tissue sample of a reed, which is a plant to be tested, with a sodium hypochlorite solution for 5min, then rinsing with sterile deionized water twice to achieve a sterilization effect, autoclaving a plant inorganic culture solution for later use, adding the sterilized plant sample into a triangular flask filled with the sterilized plant inorganic culture solution, wherein the sterilized plant inorganic culture solution contains 10mg/LPTBP, plugging a bottle opening with cotton, then culturing for two days in an oscillator, and determining the contents of PTBP and TOC in the culture solution in different periods of time;
s7: and (3) analyzing the abundance and diversity of the microorganisms: extracting total DNA of microorganisms from each soil sample by using a soil genome DNA extraction kit;
s8: and (3) enzyme activity analysis: measuring the activity of PTBP degrading enzymes of rhizosphere soil samples with plant systems and upper soil samples without plant systems, wherein the degrading enzymes are monophenol oxidase, polyphenol oxidase, monooxygenase and dioxygenase;
s9: statistical analysis: statistical analysis used SPSS software, where the analysis of the differences between two variables and the analysis of the correlations were calculated using one-way analysis of variance and pearson correlation coefficients, respectively.
2. The research method for promoting co-metabolism and degradation of alkylphenol by using phenol root exudates as claimed in claim 1, wherein the research method comprises the following steps: in the experiment, a Hoagland nutrient solution containing 10mg/LPTBP is used for feeding water, and in order to research the influence of the concentration ratio of different root exudates REs to PTBP on pollutant removal, three root exudate REs concentration gradients are set in total, so that the three root exudate REs/PTBP concentration gradients are correspondingly obtained.
3. The research method for promoting co-metabolism and degradation of alkylphenol by using phenol root exudates as claimed in claim 1, wherein the research method comprises the following steps: and (3) repeatedly operating for three periods under the concentration gradient of each root secretion REs/PTBP, operating for one period after the concentration gradient is switched every time, starting sampling after the device is stabilized, and respectively collecting the water inflow water sample with the concentration gradient of 0h and the water outflow water samples of 3 h, 12 h, 24 h and 48 h.
4. The research method for promoting co-metabolism and degradation of alkylphenol by using phenol root exudates as claimed in claim 1, wherein the research method comprises the following steps: the resulting samples were used in a series of rhizosphere effect bench tests: the soil sample is used for adsorption degradation experiments, microbial abundance and diversity analysis and enzyme activity analysis, the plant root system tissue sample is used for plant absorption experiments, and in addition, the fresh weight and the plant height of the plant are also detected to be used for calculating the plant biomass growth.
5. The research method for promoting co-metabolism and degradation of alkylphenol by using phenol root exudates as claimed in claim 1, wherein the research method comprises the following steps: adding a sterilized plant sample into a triangular flask filled with a sterilized plant inorganic culture solution, wherein the sterilized plant inorganic culture solution contains 10mg/LPTBP, plugging a bottle opening with cotton, then placing the bottle opening into an oscillator for two days, then pouring out the culture solution, taking out the plant sample, transferring the plant sample into 100mL of ultrapure water, sequentially oscillating the plant sample on an ultrasonic cleaning machine and an eddy oscillator for 1min for six times, and then measuring the content of PTBP and TOC in the obtained suspension to respectively serve as a PTBP root surface adsorption component and a TOC root surface adhesion component.
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| CN104458948A (en) * | 2014-12-05 | 2015-03-25 | 江苏省农业科学院 | Detection method of straw phenolic acid compound |
| CN111066407A (en) * | 2020-01-17 | 2020-04-28 | 河北工程大学 | Method for degrading phenolic acid substances in continuous cropping soil |
| EP3737740A1 (en) * | 2018-01-10 | 2020-11-18 | Bayer CropScience LP | Improved microbes and methods for producing the same |
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| CN104458948A (en) * | 2014-12-05 | 2015-03-25 | 江苏省农业科学院 | Detection method of straw phenolic acid compound |
| EP3737740A1 (en) * | 2018-01-10 | 2020-11-18 | Bayer CropScience LP | Improved microbes and methods for producing the same |
| CN111066407A (en) * | 2020-01-17 | 2020-04-28 | 河北工程大学 | Method for degrading phenolic acid substances in continuous cropping soil |
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