CN110184217B - Salt-tolerant denitrifying bacterium taking nitrite as nitrogen source and application thereof - Google Patents
Salt-tolerant denitrifying bacterium taking nitrite as nitrogen source and application thereof Download PDFInfo
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Abstract
The invention discloses a salt-tolerant denitrifying bacterium taking nitrite as a nitrogen source and application thereof, relates to the field of environmental microorganisms, and particularly relates to a salt-tolerant denitrifying bacterium taking nitrite as a nitrogen source. The invention aims to solve the technical problem that nitrite nitrogen in salt-containing wastewater cannot be removed because the denitrification capability of the existing denitrifying bacteria is inhibited by salt. The strain is a plant Pennella panningnanensis Hansenula phragmitetus F1, is preserved in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms, has the preservation date of 3 and 25 months in 2019 and has the preservation number of CGMCC No. 17432. The strain can denitrify under the condition of salt content and carry out denitrification. The strain is used for removing nitrite nitrogen pollutants in water.
Description
Technical Field
The invention relates to the field of environmental microorganisms, in particular to salt-tolerant denitrifying bacteria taking nitrite as a nitrogen source.
Background
With the improvement of human life quality, a large amount of high-nitrogen wastewater is generated, and high-nitrogen pollution becomes a great problem to be solved urgently at present. Among them, nitrite nitrogen existing in water has high toxicity, easily causes water deterioration, affects normal growth of aquatic animals and human beings, and even endangers life. Nitrite nitrogen pollution sources are wide, and mainly come from domestic sewage, industrial wastewater, breeding wastewater and the like. Nitrogen contaminants in domestic wastewater are converted to ammonia by microbial degradation and ammoniation, and subsequently converted to nitrite by oxidation by nitrifying bacteria, resulting in nitrite nitrogen contamination. In the industrial production process, nitrite is easy to remain as a production raw material to pollute the environment, and simultaneously, a large amount of nitrogen-containing waste water and waste residue are generated, thereby providing a material basis for the formation of nitrite. In the aquaculture process, a large amount of nitrogen pollutants are accumulated due to a plurality of factors such as accumulation of fish and shrimp excrement, overhigh stocking density, excessive feeding of residual baits and the like, and nitrogen in different forms can be converted into nitrite through the action of microorganisms, so that nitrite nitrogen in aquaculture water is increased year by year. Therefore, the removal and prevention of nitrite nitrogen pollution in water bodies is a great problem to be solved urgently.
The method for removing nitrite nitrogen in wastewater comprises a physical method, a chemical method and a biological method, wherein the biological denitrification has the advantages of high efficiency, economy, easy operation, no secondary pollution and the like and is widely researched. The removal of nitrite nitrogen is mainly achieved by nitrification and denitrification by the microorganisms. However, the pollutant components in the wastewater are complex and various, and the treatment environment is different, so that the difficulty of removing nitrite nitrogen in the wastewater is increased. For example, the salt-containing wastewater containing a large amount of nitrite nitrogen generated in people's daily life, industry and aquaculture is applied in the form of seawater substitution. The denitrification capability of common denitrifying bacteria is inhibited by salt, and nitrite nitrogen in salt-containing wastewater cannot be removed.
Disclosure of Invention
The invention provides a salt-tolerant denitrifying bacterium taking nitrite as a nitrogen source and application thereof, aiming at solving the technical problem that the denitrification capability of the existing denitrifying bacterium is inhibited by salt and can not remove nitrite nitrogen in salt-containing wastewater.
The salt-tolerant denitrifying bacterium using nitrite as nitrogen source is used as a habitat plant Panlonibacterium panningibacterium phragmitetus F1, which is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation address is No. 3 of No.1 Xilu of Beijing Kogyo Chen, the preservation date is 3 months and 25 days in 2019, and the preservation number is CGMCC No. 17432.
The strain-roosting plant Pennella panningnanensis phragmitetus F1 has homology of 99.72% with the 16S rDNA sequence of the roosting plant Pannella panningnanensis phragmitetus (Pannelobacter phragmitetus) through 16S rDNA sequence alignment analysis. The strain F1 belongs to Pannonibacter phragmitetus strain, which is determined by combining the morphological characteristics, growth conditions and physiological and biochemical identification results of thalli.
The salt-tolerant denitrifying bacteria disclosed by the invention are applied to denitrification under the salt-containing condition.
Further, the salt-containing condition is that the NaCl concentration is less than or equal to 10 g/L.
Further, the salt-containing condition is that the pH is 7-11, and NaNO is added2The initial concentration is 0.4-1.6 g/L.
Further, the inoculation amount of the salt-tolerant denitrifying bacteria is 3-7% (v/v).
The invention has the beneficial effects that:
the method screens out a strain of a habitat plant Alcaligenes pannicus (Pannonibacter phragmitetus), which has the denitrification capability of efficiently removing nitrite nitrogen; it is further found that the bacteria have certain salt tolerance and effectively exert the denitrification capability in salt-containing conditions. By utilizing the physiological and biochemical characteristics and metabolic mechanism of the bacteria with salt-tolerant denitrification capability, the problem of nitrite nitrogen removal in wastewater under the salt-containing condition can be better solved.
The Pannonibacter phragmitis F1 provided by the invention is a denitrifying bacterium using nitrite as a nitrogen source, and when the inoculation amount is 5% (v/v), the strain can treat NO2 --N、NO3 -The higher N removal rate and the large amount of gas generated in the culture process show that the Pannonibacter phragmiteus F1 has high denitrification performance. In addition, the Pannonibacter phragmitetus F1 is judged to have certain salt resistance through the salt resistance measurement result, but the growth speed is slow due to overhigh salinity. The Pannonibacter phragmiteus F1 has high-efficiency NO under the condition of NaCl salinity of 0-10 g/L2 --N、NO3 -N removal capacity, removal rate of 98% and 91% or more respectively. Therefore, the strain Pannonibacter phragmitetus F1 has good practical value and application prospect in treating high-concentration nitrite wastewater under certain salt-containing conditions.
The salt-tolerant denitrifying bacterium using nitrite as nitrogen source is used as a habitat plant Panlonibacterium panningibacterium phragmitetus F1, which is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation address is No. 3 of No.1 Xilu of Beijing Kogyo Chen, the preservation date is 3 months and 25 days in 2019, and the preservation number is CGMCC No. 17432.
The strain is used for removing nitrite nitrogen pollutants in water.
Drawings
FIG. 1 is a photograph showing the gas generation phenomenon during the culture of Pannonibacter phragmitetus F1 according to the present invention;
FIG. 2 is an electrophoretogram of PCR amplification product of genomic DNA of Pannonibacter phragmitetus F1 of the present invention; in the figure, F1 is the strain Pannonibacter phragmitetus F1;
FIG. 3 is a phylogenetic tree diagram constructed from the 16S rDNA gene sequences of Pannonibacter phragmitetus F1 and similar strains according to the present invention;
FIG. 4 shows an example of Pannonibacter phragmitetus F1 vs NO2 --N and NO3 --N removal effect map; wherein a represents NO2 --N, b represents NO3 --N；
FIG. 5 is a graph of the Pannonibacter phragmitetus F1 vs NO at different salinity2 --N and NO3 --a graph of the removal capacity versus N; wherein ■ represents NO2 --N, ● represents NO3 --N。
Detailed Description
The technical solution of the present invention is not limited to the specific embodiments listed below, and includes any combination of the specific embodiments.
The first embodiment is as follows: in the embodiment, a salt-tolerant denitrifying bacterium taking nitrite as a nitrogen source is taken as a habitat plant Panlonibacterium panningibacterium phragmiteus F1, which is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation address is No. 3 of No.1 Xilu of the sunward area of Beijing, the preservation date is 3 months and 25 days in 2019, and the preservation number is CGMCC No. 17432.
The colony of the plant Pennella panningibacter phragmiteus F1 is in a round point shape, is extremely small, has neat edges, and has smooth and glossy surface. The rod shape of the bacteria is observed under a microscope, and the bacteria are gram negative bacteria.
The physiological and biochemical characteristics of Pannonibacter phragmitetus F1 in this embodiment are shown in table 1:
TABLE 1 physio-biochemical characteristics of Pannonibacter phragmitetus F1
+: growth or positive; -: no growth or negative.
The second embodiment is as follows: the screening method of the Alcaligenes pannicus Pannonibacter phragmitetus F1 is realized by the following steps:
the method comprises the following steps: sucking 2mL of seawall muddy water samples collected from the Mandarin city of Dalian city of Liaoning province (northern latitude 39 degrees 38 '31' east longitude 122 degrees 58 '19'), placing the samples into a 250mL conical flask, and adding 200mL of strain denitrification culture medium; sealing the bottle mouth by using a sealing film to manufacture an anoxic environment; standing and culturing at the constant temperature of 30 ℃ for 5-7 days to obtain an enrichment culture solution;
step two: performing gradient dilution on the enriched culture solution obtained in the step one by adopting a multiple dilution method, respectively sucking 0.5mL of each dilution suspension into a strain screening plate culture medium (which is placed overnight and does not grow with mixed bacteria), uniformly coating the enriched culture solution by using a glass triangular coating rod, pouring the enriched culture solution into a second layer of the sterilized screening plate culture medium with the temperature not higher than 40 ℃, and manufacturing an anoxic environment for culturing denitrifying bacteria; sealing the periphery of the flat plate by using a sealing film; inverting the plate after the second layer of culture medium is solidified, putting the plate into a constant temperature incubator at the temperature of 30 ℃, and culturing until obvious bacterial colonies grow out; selecting the separated strain, streaking and purifying the strain for multiple times on a double-layer strain screening plate culture medium until no mixed bacteria is observed under a microscope, then inoculating the strain into 200mL of denitrification culture medium, and standing and expanding the culture at constant temperature under the anoxic condition of 30 ℃;
step three: inoculating the bacterial liquid obtained by the enlarged culture in the step two into 200mL of fresh bacterial strain denitrification culture medium by the inoculation amount of 5%, culturing for 5 days under the anoxic condition at the temperature of 30 ℃, and observing the turbidity condition of the culture medium and whether gas is generated in the culture process; then using NO in the denitrifying culture medium of the strain2 --N、NO3 -Screening by using-N removal rate as an index, and selecting cultured NO2 --N、NO3 -And (4) separating and purifying the bacterial liquid with high-N removal rate (the removal rate is more than 84%) to obtain a strain with a high-efficiency denitrification function, namely completing the screening of the salt-tolerant denitrifying bacteria taking nitrite as a nitrogen source.
Wherein the denitrification medium (/ L) of the strain consists of 5g of CH3COONa, 1g of K2HPO40.8g of NaNO20.03g of CaCl21g of NaCO30.06g of FeSO4·7H2O, 0.2g MgSO4·7H2O and 1000mL of deionized water, wherein the pH is natural, and the mixture is sterilized for 30min under the high-pressure condition of the temperature of 121 ℃;
the strain screening plate culture medium (/ L) is prepared by adding 20g of agar per liter based on the strain denitrification culture medium, and sterilizing at 121 deg.C under high pressure for 30min under natural pH.
Identification of Pannonibacter phragmitetus F1
16S rDNA sequencing and evolutionary tree making:
extraction of total genomic DNA of strain F1: culturing strain, collecting thallus, cracking thallus, extracting DNA, depositing, washing, pumping out (drying) to obtain total genome DNA, taking 5 microliter, and detecting with 1% agarose gel electrophoresis.
PCR amplification of strain F1 genomic DNA: taking total DNA as a template, and taking an amplification primer as a pair of universal primers;
the forward primer was 27F: 5'-AGAGTTTGATCCTGGCTCAG-3', respectively;
the reverse primer is 1492R: 5'-GGTTACCTTGTTA CGACTT-3' are provided.
PCR amplification was performed using the genomic DNA as a template. The PCR reaction system is as follows:
the PCR reaction was carried out in a 25. mu.L system. The reaction system comprises the following components: template DNA 0.5. mu. L, dNTP (mix) 1. mu.L, Taq Buffer (with MgCl. RTM. MgCl)2)2.5 μ L, Taq enzyme 0.2 μ L, primer F0.5 μ L, primer R0.5 μ L, and double distilled water to 25 μ L.
PCR amplification requires pre-denaturation at 94 ℃ for 4min, denaturation at 94 ℃ for 45s, annealing at 55 ℃ for 45s, extension at 72 ℃ for 1min, 30 cycles in total, repair extension at 72 ℃ for 10min, termination of the reaction at 4 ℃, and the obtained fragments are used for sequencing.
The 16S rDNA of the Pannonibacter phragmitetus F1 is shown as SEQ ID NO: 1 is shown.
mu.L of PCR amplification product was detected by 1% agarose gel electrophoresis.
The electrophoretogram of the PCR amplification product is shown in FIG. 2.
After the verification, the gel strip was cut off, and the PCR product was purified using a DNA gel recovery kit (Shanghai Bioengineering Co., Ltd.).
Phylogenetic Tree As shown in FIG. 3, it can be seen from FIG. 3 that the strain F1 has 99.72% homology with the 16S rDNA sequence of the Thermus pennelliniella (Pannonibacter phragmitetus) plant.
According to the ' handbook of identifying common bacteria system ' and ' Bergey ' bacteria identification handbook ', physiological and biochemical characteristics and 16S rDNA sequencing are combined, the strain F1 is identified as the plant Alcaligenes panlongensis (Pannonibacter phragmitetus), and the homology of the 16S rDNA sequence is 99.72%. The named strain F1 was: the Pannonibacter phragmitetus F1 has a preservation date of 2019, 3 and 25 months, the preservation address is the China general microbiological culture Collection center of the culture Collection of microorganisms, and the preservation number is CGMCC No: 17432.
the third concrete implementation mode: the determination of the salt tolerance of the habitat of the plant Alcaligenes pannini Pannonibacter phragmitetus F1 is as follows:
preparing screening plate culture media with different salinity gradients, wherein the NaCl concentrations are respectively 0g/L, 30g/L, 50g/L, 70g/L and 100 g/L; the Pannonibacter phragmiteus F1 is inoculated to a culture medium plate with various salinity by streaking, and is put into a constant temperature incubator at 30 ℃ for culture for 5 days, and whether the strain grows or not is taken as the basis of whether the strain resists the corresponding salinity or not; the culture medium without denitrifying bacteria was used as a blank, and 3 cells were used in parallel.
Table 2 shows the growth of Pannonibacter phragmitetus F1 in different salinity gradient media. After 1d of culture, no bacterial colony appears in each salinity culture medium; after 2d culture, obvious colony growth is carried out on a culture medium containing 0g/L and 30g/L NaCl, a trace amount of colony growth is carried out on a culture medium containing 50g/L NaCl, and sterile colony growth is carried out on a culture medium containing 70g/L and 100g/L NaCl; after 3d culture, obvious bacterial colonies appear on a culture medium containing 50g/L NaCl, and micro bacterial colonies grow on the culture medium containing 70g/L NaCl; after 5 days of culture, colonies grew on the medium containing 0g/L, 30g/L, 50g/L and 70g/L NaCl, and no colony growth was observed on the medium containing 100g/L NaCl. Pannonibacter phragmitetus F1 is proved to have certain salt tolerance, but too high salinity can cause slow growth speed and even inhibit growth.
TABLE 2 Pannonibacter phragmitetus F1 growth in different salinity gradient media
++: the appearance of a clear colony on the surface of the medium indicates that Pannonibacter phragmitetus F1 grows normally;
+: micro-colonies appear on the surface of the culture medium, which shows that the Pannonibacter phragmitetus F1 has a growth phenomenon, but the growth speed is slow;
-: no colonies appeared on the surface of the medium, indicating that Pannonibacter phragmitetus F1 did not grow.
The fourth concrete implementation mode: the denitrification performance of the inhabitant plant Panlonibacterium panningnieri Panronibacter phragmitetus F1 is determined as follows:
preparing denitrification culture media with different salinity gradients, wherein the NaCl concentrations are respectively 0g/L, 10g/L, 30g/L, 50g/L, 70g/L and 100 g/L; transferring the strain to sterilized culture medium with various salinity according to the inoculation amount of 5%, sealing the bottle mouth, and performing static culture at constant temperature of 30 ℃ for 5 d; determination of NO in the Medium2 --N and NO3 -N concentration, the denitrification performance of the strain is tested. The culture medium without denitrifying bacteria was used as a blank, and 3 cells were used in parallel.
After 5 days of culture, Pannonibacter phragmitetus F1 has certain NO under various salinity conditions2 --N、NO3 --N removal, but too high salinity inhibits strain growth, thereby reducing its NO tolerance2 --N、NO3 --N removal capacity. The Pannonibacter phragmiteus F1 has high-efficiency NO under the condition of NaCl salinity of 0-10 g/L2 --N、NO3 -The N removal capacity is high, and the removal rate is respectively over 98% and 91%. Under the condition of NaCl salinity of 30-100 g/L, the excessive salinity inhibits the growth of the strain, so that the Pannonibacter phragmiteus F1 is used for treating NO2 --N、NO3 -the-N removal capacity is significantly reduced.
The following examples were used to demonstrate the beneficial effects of the present invention:
example one
Culture conditions of the strains:
the Pannonibacter phragmitetus F1 preserved on the preservation inclined tube culture medium is scraped by an inoculating loop to obtain 2-ring lawn, the 2-ring lawn is inoculated into a 250mL conical flask filled with 200mL of sterilized denitrification culture medium, the opening of the conical flask is sealed, and the conical flask is placed in a constant temperature incubator at 30 ℃ for 5 days to obtain fresh enriched bacterial liquid which is used as seed liquid. In the experiment, the seed solution is inoculated in the denitrification culture medium or the wastewater according to the inoculation amount of 5 percent.
Denitrification performance test of the strain:
enrichment culture of Pannonibacter phragmitetus F1, sealing the bottle, and standing at 30 deg.C for cultureThe culture medium is turbid and generates gas to be used as seed liquid; inoculating the mixture into 200mL of sterilized denitrification culture medium with an inoculation amount of 5%, filling the mixture into a 250mL conical flask, and placing the conical flask at a constant temperature of 30 ℃ for sealing and standing culture for 5 d; taking a fixed amount of culture solution at regular time, and measuring NO in the culture solution2 --N and NO3 -N, detecting the denitrification performance of the strain. The culture medium without denitrifying bacteria was used as a blank, and 3 cells were used in parallel. As a result, as shown in FIG. 4, NO in the denitrifying medium after 12 hours of cultivation3 --N and NO3 -The N concentration increases markedly, then decreases gradually; after 48h incubation, Pannonibacter phragmitetus F1 for NO2 --N、NO3 -The removal rates of-N were 50.79% and 37.76%, respectively; after 96h of incubation, NO in the culture broth3 -The concentration of-N is reduced from the initial 212.33 + -5.95 mg/L to 9.67 + -1.03 mg/L when NO is present3 -The removal rate of-N reaches a maximum of 95.45%; after 120h of culture, NO in the culture broth2 -The concentration of-N is reduced from the initial 579.50 + -23.13 mg/L to 7.17 + -0.62 mg/L when NO is present2 -The removal of-N reached a maximum of 98.76%.
Determination of oxygen demand of strains
Cooling the sterilized test tube filled with the screening separation culture medium to about 40 ℃, taking a small amount of bacterial colonies of the strain to be tested by using an inoculating needle, performing vertical puncture inoculation on the bottom of the test tube, and immediately placing the test tube in cold water for cooling and solidification. After coagulation, the tubes were placed upright in a 30 ℃ incubator for 5 days. The culture medium without denitrifying bacteria was used as a blank, and 3 cells were used in parallel. After culturing for 5 days, the Pannonibacter phragmiteus F1 grows in the culture medium along the puncture straight line, and the Pannonibacter phragmiteus F1 can be preliminarily judged to be facultative anaerobe.
Influence of salinity on the denitrifying Properties of the strains
Denitrifying culture mediums with different salinity gradients are prepared, namely NaCl concentrations of 0g/L, 10g/L, 30g/L, 50g/L, 70g/L and 100g/L respectively. The Pannonibacter phragmitetus F1 was inoculated into each salinity medium at 5% (v/v),sealing the bottle mouth, and standing and culturing at constant temperature of 30 ℃ for 5 d. Determination of NO in culture broth2 --N、NO3 -N concentration, the denitrification performance of Pannonibacter phragmitetus F1 was tested. The culture medium without denitrifying bacteria was used as a blank, and 3 cells were used in parallel. As shown in FIG. 5, Pannonibacter phragmitetus F1 for NO2 --N、NO3 -The removal ability of-N is significantly affected by salinity. After 5 days of culture, Pannonibacter phragmiteus F1 was cultured in denitrifying culture medium containing 0g/L and 10g/L NaCl for NO2 --N、NO3 -Significant removal of-N, NO2 --N、NO3 -the-N removal rate is respectively above 98% and 91%, which indicates that Pannonibacter phragmitetus F1 normally grows under the condition of containing 10g/L NaCl, and NO2 --N、NO3 -the-N removal capacity is not affected by salinity. Pannonibacter phragmitetus F1 for NO in culture broth containing 30g/L, 50g/L and 70g/L NaCl2 --N、NO3 -The removing capability of N is poor, the removing rate is in the range of 9-35%, which shows that Pannonibacter phragmiteus F1 grows slowly under the condition of containing 50-70 g/L NaCl, and NO is2 --N、NO3 -the-N removal capacity is significantly reduced by salinity. Pannonibacter phragmitetus F1 for NO in a culture medium containing 100g/L NaCl2 --N and NO3 -The removal rate of-N is about 9 percent, which indicates that the Pannonibacter phragmitetus F1 is inhibited from growing under the condition of containing 100g/L NaCl, and NO is2 --N、NO3 -the-N removal capability is severely affected.
Optimum salt-tolerant denitrification condition test of strain
200mL of sterilized denitrification medium containing 10g/L NaCl was placed in a 250mL conical flask, and the Pannonibacter phragmiteus F1 strain of the present invention was cultured in a sealed and static manner at 30 ℃ to examine NaNO at different pH values (3, 5, 7, 9, 10, 11)2Initial concentrations (0.4g/L, 0.8g/L, 1.6g/L, 2.4g/L, 3.2g/L) and inoculum size (v/v) (1%, 3%, 5%, 7%, 10%) of Pannonibacter phragmitetus F1 pairsNO2 --N、NO3 --N removal capacity. The result shows that the Pannonibacter phragmiteus F1 can adapt to wide neutral and alkaline environments under the condition of 10g/L NaCl salinity, and the Pannonibacter phragmiteus F1 can be used for NO under the condition that the pH is 7-112 -The N division rates are all over 99 percent; pannonibacter phragmitetus F1 for NO in alkaline environment3 -The removal rate of-N is within the range of 48-83%, and NO is obtained when pH is 103 -The highest N removal rate was 83.17%. The best NaNO of Pannonibacter phragmitetus F12The initial concentration is 0.4-1.6 g/L, and the Pannonibacter phragmitetus F1 is used for NO2 --N、NO3 -The removal rate of-N reaches more than 99% and 82% respectively. The optimal inoculation amount of Pannonibacter phragmitetus F1 is 3-7% (v/v), and the strain F2 is used for NO2 --N、NO3 -The removal rate of-N reaches more than 99% and 89% respectively.
Sequence listing
<110> institute of geography and agroecology of northeast China academy of sciences
<120> one strain of salt-tolerant denitrifying bacteria using nitrite as nitrogen source and application thereof
<160> 3
<210> 1
<211> 1440
<212> DNA
<213> the plant Alcaligenes panonensis (Pannonibacter phragmitetus)
<220>
<223> Alcaligenes penlongus (Pannonibacter phragmitetus) F1
<400> 1
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cccgcaaggg gagtggcaga cgggtgagta acacgtggga acataccctt tggtgcggaa 120
caacagctgg aaacggctgc taataccgca tgcgccctac gggggaaaga tttatcgccg 180
aaggattggc ccgcgtctga ttagctagtt ggtgaggtaa tggctcacca aggcgacgat 240
cagtagctgg tctgagagga tgatcagcca cactgggact gagacacggc ccagactcct 300
acgggaggca gcagtgggga atattggaca atgggcgcaa gcctgatcca gccatgccgc 360
gtgagtgatg aaggccctag ggttgtaaag ctctttcagc gaggaggata atgacgttac 420
tcgcagaaga agccccggct aacttcgtgc cagcagccgc ggtaatacga agggggctag 480
cgttgttcgg aatcactggg cgtaaagcgc acgtaggcgg acttttaagt caggggtgaa 540
atcccggggc tcaacctcgg aactgccttt gatactggaa gtctggagtc cgagagaggt 600
gagtggaact ccgagtgtag aggtgaaatt cgtagatatt cggaagaaca ccagtggcga 660
aggcggctca ctggctcggt actgacgctg aggtgcgaaa gcgtggggag caaacaggat 720
tagataccct ggtagtccac gccgtaaacg atggaagcta gccgtcaggg tgcatgcatc 780
ttggtggcgc agttaacgca ttaagcttcc cgcctgggga gtacggtcgc aagattaaaa 840
ctcaaaggaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa 900
cgcgcagaac cttaccagct cttgacattt ggtgctacca cgggagaccg tgggttccct 960
tcggggacgc caggacaggt gctgcatggc tgtcgtcagc tcgtgtcgtg agatgttggg 1020
ttaagtcccg caacgagcgc aaccctcgcc cttagttgcc agcattaagt tgggcactct 1080
agggggactg ccggtgataa gccgagagga aggtggggat gacgtcaagt cctcatggcc 1140
cttacgggct gggctacaca cgtgctacaa tggcggtgac agtgggcagc gaaggggtga 1200
cccggagcta atctccaaaa gccgtctcag ttcggattgt tctctgcaac tcgagagcat 1260
gaagttggaa tcgctagtaa tcgcgtaaca gcatgacgcg gtgaatacgt tcccgggcct 1320
tgtacacacc gcccgtcaca ccatgggagt tgggtttacc cgaaggtggt gcgctaaccg 1380
caaggaggca gccaaccacg gtaggctcag cgactggggt gaagtcgtaa caaggtaacc 1440
<210>2
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> Forward primer 27F
<400>2
<210>3
<211> 19
<212> DNA
<213> Artificial sequence
<220>
<223> reverse primer 1492R
<400>3
ggttaccttgttacgactt 19
Claims (5)
1. A salt-tolerant denitrifying bacterium using nitrite as nitrogen source is characterized in that: the strain is a plant Pennella pennyensis F1, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, has the preservation address of No. 3 Xilu No.1 of Beijing Kogyo Chen, 3 months and 25 days in 2019 and the preservation number of CGMCC No. 17432.
2. Use of the salt-tolerant denitrifying bacteria of claim 1 to denitrify nitrogen under salt-containing conditions.
3. Use according to claim 2, characterized in that the salt-containing conditions are a NaCl concentration ≤ 10 g/L.
4. The use according to claim 2, wherein the denitrification reaction is carried out under a pH of 7 to 11 and with NaNO2The initial concentration is 0.4-1.6 g/L.
5. The use according to claim 2, characterized in that the salt-tolerant denitrifying bacteria are inoculated in an amount of 3-7% (v/v).
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| CN106834184A (en) * | 2017-03-03 | 2017-06-13 | 中南大学 | A kind of composite flora and its application in Cr (VI) contaminated soil remediation |
| CN107309270A (en) * | 2017-06-23 | 2017-11-03 | 河南大学 | Application of one plant of BB bacterium in reduction red mud pH value |
| WO2018213568A1 (en) * | 2017-05-17 | 2018-11-22 | President And Fellows Of Harvard College | Biofertilzer and methods of making and using same |
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| CN106834184A (en) * | 2017-03-03 | 2017-06-13 | 中南大学 | A kind of composite flora and its application in Cr (VI) contaminated soil remediation |
| WO2018213568A1 (en) * | 2017-05-17 | 2018-11-22 | President And Fellows Of Harvard College | Biofertilzer and methods of making and using same |
| CN107309270A (en) * | 2017-06-23 | 2017-11-03 | 河南大学 | Application of one plant of BB bacterium in reduction red mud pH value |
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| High-efficiency inorganic nitrogen removal by newly isolated Pannonibacter phragmitetus B1;Hong Bai等;《Bioresource Technology 271》;20180918;第271卷;第91-99页 * |
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| 以聚丁酸丁二醇酯为碳源去除含盐水体硝酸盐的研究;罗国芝等;《环境污染与防治》;20130330;第35卷(第3期);参见第20-25页 * |
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