CN113234625A - Acinetobacter pittericus and application thereof - Google Patents
Acinetobacter pittericus and application thereof Download PDFInfo
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Abstract
The invention discloses acinetobacter cutaneus and application thereof, and relates to acinetobacter cutaneus and application thereof. The invention aims to provide an acinetobacter cutaneus for degrading 2,2 ', 4, 4' -tetrabromobisphenol, the acinetobacter cutaneus is acinetobacter cutaneus (Acinetobacter pittii) GB-2, and is preserved in China center for type culture collection, the preservation address is Wuhan university in Wuhan City, China, the preservation date is 1 month and 19 days in 2021 year, and the preservation number is CCTCCNO.M 2021107. The screened strain acinetobacter cutaneus GB-2 can be used for bioremediation of water bodies and soil polluted by BDE-47, and has high application value. The invention is applied to the technical field of environmental microorganisms.
Description
Technical Field
The invention relates to acinetobacter cutaneus and application thereof.
Background
Polybrominated diphenyl ethers (PBDEs) are one of persistent organic pollutants, are widely applied to various industries as an important additive flame retardant, such as the fields of plastics, electronic appliances, furniture, interior decoration, traffic, petroleum, mining and the like, and are detected in various environmental media. The PBDEs share 209 homologs, and the 2,2 ', 4, 4' -tetrabromobisphenol (BDE-47) is one of the PBDEs which are the most widely distributed, have the highest content in organisms and have the highest toxicity to human bodies and animals, although not the PBDEs homolog with the highest content in the environment. BDE-47 not only has high lipophilicity, persistence, bioaccumulation and mobility, but also has toxic effect on organisms as food chains are enriched in organisms and human bodies. Therefore, research on biodegradation of BDE-47 can accelerate degradation, and has important significance for protecting environmental health and reducing human safety risks.
Due to the long-term use of BDE-47 and the illegal disassembly of various electronic wastes containing BDE-47 in recent years, the BDE-47 can permeate into soil along with precipitation and surface runoff in modes of volatilization, sedimentation and the like. At present, the methods mainly adopted for degrading BDE-47 at home and abroad comprise a photodegradation method, a plant degradation method, a biodegradation method and a zero-valent iron reduction method, and in the research process of various degradation technical methods, the microbial degradation is considered to be one of the most promising means at present due to the characteristics of remarkable effect, low technical cost, no secondary pollution and the like.
The microbial degradation refers to the conversion of macromolecular organic matters into small molecular compounds and the complete decomposition of the organic matters into carbon dioxide and water under the action of microorganisms, and is considered as a main way for removing persistent organic pollutants in the environment. In the microorganisms obtained at present, the number of microorganisms capable of degrading BDE-47 is small, the degradation effect is not obvious, and aiming at the problem, the screening of the efficient degrading bacteria of BDE-47 is a hot problem of environmental management and repair research on BDE-47 pollution, so that a foundation is laid for reducing and eliminating the BDE-47 pollution soil problem.
Disclosure of Invention
The invention aims to provide an acinetobacter cutaneus for degrading 2,2 ', 4, 4' -tetrabromobisphenol.
The Acinetobacter pitteus strain is Acinetobacter pittii GB-2, is preserved in China center for type culture collection, is preserved at Wuhan university in Wuhan City, China, has the preservation date of 2021 year, 1 month and 19 days, and has the preservation number of CCTCC No. M2021107.
The invention discloses an application of acinetobacter cutaneus in degrading environmental pollutants, wherein the environmental pollutants are 2,2 ', 4, 4' -tetrabromobiphenyl ether.
The invention adopts the electronic dismantling of BDE-47 polluted soil in the industrial park of Taizhou area of Taizhou, Zhejiang, takes fresh soil (the sampling depth is 0-20cm), removes impurities in the sample in the collection process, places the sample in a sterile bag, places the sample in an ice box, brings the sample back to a laboratory, and preserves the sample in a refrigerator at 4 ℃ for traditional microbial culture. Domestication, separation and screening of BDE-47 aerobic degradation strains:
sieving soil collected from an electronic waste dismantling field, weighing 10g of the soil, placing the soil into a conical flask with glass beads, adding a strain screening culture medium taking BDE-47 as a unique carbon source, and culturing the soil in a shaking culture box at 35 ℃ and 150rpm to ensure that bacteria are dispersed in liquid in a single cell state and fully contact with nutrient substances;
absorbing 10mL of the bacterial suspension in the conical flask, transferring the bacterial suspension to 90mL of newly-configured BDE-47 strain screening culture medium, carrying out subculture under the same conditions, repeating the steps, and carrying out culture for several cycles;
thirdly, acclimating strains by adopting a pollutant concentration gradient acclimation method: the concentration of BDE-47 in the bacterial domestication system is increased according to 10 mug/L, 20 mug/L, 50 mug/L and 100 mug/L in turn, 7 days is a domestication period, and the domestication period is four periods in total;
fourthly, performing strain domestication by adopting a maximum pollutant concentration domestication method: the concentration of the BDE-47 of the bacterial domestication system is 100 mug/L, 7d is one domestication period, and the domestication period is four periods in total, so that a flora which can grow by taking the BDE-47 as a unique carbon source is obtained;
fifthly, absorbing 1mL of bacterial liquid in the last period of screening culture medium of the degrading bacterial strain, and diluting the obtained bacterial liquid to 10 times by a 10-fold dilution method-1-10-6Coating 100 μ L of gradient liquid on LB solid culture medium, uniformly coating with sterilizing coating rod, and placing in 37 deg.C constant temperature incubatorCulturing;
after independent colonies grow out, observing morphological characteristics of each colony for screening, selecting the screened colony by using an inoculating loop, carrying out streaking separation on a beef extract solid culture medium, and carrying out repeated streaking purification until a single colony is screened to obtain a pure bacterium;
seventhly, on the basis of primary screening, re-screening the bacterial strains with better degradation efficiency according to whether the bacterial strains have higher BDE-47 degradation rate or not, and naming the bacterial strains as GB-2 and identifying the bacterial strains. Under the condition of aseptic operation, diluting the bacterial suspension with sterile water, putting 100 mu L of bacterial liquid on an LB solid culture medium flat plate, uniformly coating the bacterial liquid with a sterilized coating rod, inversely placing the bacterial liquid in a constant-temperature incubator at 37 ℃ for culturing until a single bacterial colony grows out, wherein the bacterial colony is milky white, round, smooth and moist in surface, free of wrinkles, neat in edge and easy to pick up; the cell shape is short rod shape; gram stain negative; the physiological and biochemical characteristics of the method are positive catalase reaction, negative oxidase reaction, positive citric acid utilization, positive starch hydrolysis, negative methyl red test reaction, negative nitrate reduction reaction, negative VoP test reaction and positive indole production test. Extracting bacterial DNA, carrying out PCR amplification on the obtained bacterial genome DNA by using bacterial 16S rDNA universal primers, (27F: 5'-AGAGTTTGATCCTGGCTCAG-3', 1492R: 5'-GGTTACCTTGTTACGACTT-3'), tapping after electrophoresis to recover PCR products, entrusting 16S rDNA sequencing by Jinzhi Biotechnology Limited, Suzhou, carrying out homology comparison on sequencing results and 16S rDNA sequences registered in Genbank by using BLAST software, finding that the homology of the strain GB-2 obtained by screening the embodiment and the Pitotobacter (Acinetobacter pittii) reaches 100%, submitting the sequences to Genbank database, and obtaining the registration number of MT 150630. According to the sequencing result of 16S rDNA, a homologous sequence is found in a GenBank database by using Blast tools provided by NCBI and related software such as MEGA7.0 and the like, a phylogenetic tree is established, the homology of the strain and the Acinetobacter cutaneus is the highest and reaches 100%, and the strain is finally named as Acinetobacter cutaneus (Acinetobacter pittii) GB-2.
The screened strain acinetobacter cutaneus GB-2 is subjected to shaking culture at the temperature of 35 ℃ and 150rpm for 7 days at the pH value of 7.0, the degradation rate of BDE-47 with the initial concentration of 100 mu g/L can reach more than 70%, secondary pollution to the environment is reduced, and the strain can be used for bioremediation of water bodies and soil polluted by BDE-47 and has high application value.
Drawings
FIG. 1 is a colony morphology of Acinetobacter pitteus GB-2 plate;
FIG. 2 shows a phylogenetic tree of strain GB-2.
Detailed Description
The first embodiment is as follows: the Acinetobacter pitteus strain is Acinetobacter pittii (Acinetobacter pittii) GB-2 which is preserved in China center for type culture Collection, wherein the preservation address is Wuhan university in Wuhan City, China, the preservation date is 2021, 1 and 19 days, and the preservation number is CCTCC No. M2021107.
In the embodiment, BDE-47 polluted soil in an industrial park is electronically disassembled by adopting the mountain river town of the bridge area of Taizhou city of Zhejiang province, fresh soil (0-20cm) is taken, impurities in a sample are removed in the collection process, the sample is placed in a sterile bag, an ice box is placed into the sterile bag, and the sterile bag is brought back to a laboratory and stored in a refrigerator at 4 ℃ for traditional microbial culture. Domestication, separation and screening of BDE-47 aerobic degradation strains:
sieving soil collected from an electronic waste dismantling field, weighing 10g of the soil, placing the soil into a conical flask with glass beads, adding a strain screening culture medium taking BDE-47 as a unique carbon source, and culturing the soil in a shaking culture box at 35 ℃ and 150rpm to ensure that bacteria are dispersed in liquid in a single cell state and fully contact with nutrient substances;
absorbing 10mL of the bacterial suspension in the conical flask, transferring the bacterial suspension to 90mL of newly-configured BDE-47 strain screening culture medium, carrying out subculture under the same conditions, repeating the steps, wherein 7d is a period, and carrying out culture for 4 periods;
thirdly, acclimating strains by adopting a pollutant concentration gradient acclimation method: the concentration of BDE-47 in the bacterial domestication system is increased according to 10 mug/L, 20 mug/L, 50 mug/L and 100 mug/L in turn, 7 days is a domestication period, and the domestication period is four periods in total;
fourthly, performing strain domestication by adopting a maximum pollutant concentration domestication method: the concentration of the BDE-47 of the bacterial domestication system is 100 mug/L, 7d is one domestication period, and the domestication period is four periods in total, so that a flora which can grow by taking the BDE-47 as a unique carbon source is obtained;
fifthly, absorbing 1mL of bacterial liquid in the last period of screening culture medium of the degrading bacterial strain, and diluting the obtained bacterial liquid to 10 times by a 10-fold dilution method-1-10-6Coating 100 mu L of the gradient liquid on an LB solid culture medium, uniformly coating the gradient liquid by using a sterilization coating rod, and culturing in a constant-temperature incubator at 37 ℃;
after independent colonies grow out, observing morphological characteristics of each colony for screening, selecting the screened colony by using an inoculating loop, carrying out streaking separation on a beef extract solid culture medium, and carrying out repeated streaking purification until a single colony is screened to obtain a pure bacterium;
seventhly, on the basis of primary screening, re-screening the bacterial strains with better degradation efficiency according to whether the bacterial strains have higher BDE-47 degradation rate or not, and naming the bacterial strains as GB-2 and identifying the bacterial strains. Under the condition of aseptic operation, diluting the bacterial suspension with sterile water, taking 100 mu L of bacterial liquid on a yeast extract solid culture medium flat plate, uniformly coating the bacterial liquid with a sterilized coating rod, inversely placing the bacterial liquid in a constant-temperature incubator at 37 ℃ for culturing until a single bacterial colony grows out, wherein the bacterial colony is milky white and round, has a smooth and moist surface, does not have wrinkles, and is neat in edge and easy to pick up; the cell shape is short rod shape; gram stain negative; the physiological and biochemical characteristics of the method are positive catalase reaction, negative oxidase reaction, positive citric acid utilization, positive starch hydrolysis, negative methyl red test reaction, negative nitrate reduction reaction, negative VoP test reaction and positive indole production test. Extracting bacterial DNA, carrying out PCR amplification on the obtained bacterial genome DNA by using bacterial 16S rDNA universal primers, (27F: 5'-AGAGTTTGATCCTGGCTCAG-3', 1492R: 5'-GGTTACCTTGTTACGACTT-3'), tapping after electrophoresis to recover PCR products, entrusting 16S rDNA sequencing by Jinzhi Biotechnology Limited, Suzhou, carrying out homology comparison on sequencing results and 16S rDNA sequences registered in Genbank by using BLAST software, finding that the homology of the strain GB-2 obtained by screening the embodiment and the Pitotobacter (Acinetobacter pittii) reaches 100%, submitting the sequences to Genbank database, and obtaining the registration number of MT 150630. According to the sequencing result of 16S rDNA, a homologous sequence is found in a GenBank database by using Blast tools provided by NCBI and related software such as MEGA7.0 and the like, a phylogenetic tree is established, the homology of the strain and the Acinetobacter cutaneus is the highest and reaches 100%, and the strain is finally named as Acinetobacter cutaneus (Acinetobacter pittii) GB-2.
The second embodiment is as follows: the application of acinetobacter cutaneus in degrading environmental pollutants is provided, wherein the environmental pollutants are 2,2 ', 4, 4' -tetrabromobisphenol.
The screened strain acinetobacter cutaneus GB-2 is subjected to shaking culture at the temperature of 35 ℃ and 150rpm for 7 days at the pH value of 7.0, the degradation rate of BDE-47 with the initial concentration of 100 mu g/L can reach over 70 percent, secondary pollution to the environment is reduced, and the strain can be used for bioremediation of water bodies and soil polluted by BDE-47 and has high application value.
The third concrete implementation mode: the screening method of Acinetobacter pinteus (Acinetobacter pittii) GB-2 in the embodiment comprises the following steps: electronic disassembling of BDE-47 polluted soil in industrial park by adopting mountain river town of Taiwan bridge area of Taizhou city of Zhejiang, taking (0-20cm) fresh soil, removing impurities in sample in collection process, placing in sterile bag, placing in ice box, taking back to laboratory, preserving in refrigerator at 4 deg.C, and using for traditional microorganism culture. Domestication, separation and screening of BDE-47 aerobic degradation strains:
sieving soil collected from an electronic waste dismantling field, weighing 10g of the soil, placing the soil into a conical flask with glass beads, adding a strain screening culture medium taking BDE-47 as a unique carbon source, and culturing the soil in a shaking culture box at 35 ℃ and 150rpm to ensure that bacteria are dispersed in liquid in a single cell state and fully contact with nutrient substances;
absorbing 10mL of the bacterial suspension in the conical flask, transferring the bacterial suspension to 90mL of newly-configured BDE-47 strain screening culture medium, carrying out subculture under the same conditions, repeating the steps, and carrying out culture for 4 periods;
thirdly, acclimating strains by adopting a pollutant concentration gradient acclimation method: the concentration of BDE-47 in the bacterial domestication system is increased according to 10 mug/L, 20 mug/L, 50 mug/L and 100 mug/L in turn, 7 days is a domestication period, and the domestication period is four periods in total;
fourthly, performing strain domestication by adopting a maximum pollutant concentration domestication method: the concentration of the BDE-47 of the bacterial domestication system is 100 mug/L, 7d is one domestication period, and the domestication period is four periods in total, so that a flora which can grow by taking the BDE-47 as a unique carbon source is obtained;
fifthly, absorbing 1mL of bacterial liquid in the last period of screening culture medium of the degrading bacterial strain, and diluting the obtained bacterial liquid to 10 times by a 10-fold dilution method-1-10-6Coating 100 mu L of the gradient liquid on an LB solid culture medium, uniformly coating the gradient liquid by using a sterilization coating rod, and culturing in a constant-temperature incubator at 37 ℃;
after independent colonies grow out, observing morphological characteristics of each colony for screening, selecting the screened colony by using an inoculating loop, carrying out streaking separation on a beef extract solid culture medium, and carrying out repeated streaking purification until a single colony is screened to obtain a pure bacterium;
seventhly, on the basis of primary screening, re-screening the bacterial strains with better degradation efficiency according to whether the bacterial strains have higher BDE-47 degradation rate or not, and naming the bacterial strains as GB-2 and identifying the bacterial strains.
Wherein the main experimental reagents comprise: BDE-47 Standard (>99%), n-hexane (chromatographically pure), methanol (chromatographically pure), beef extract, peptone, sodium chloride, agar, yeast extract, Na2HPO4·2H2O，Ca(NO3)2·4H2O，(NH4)2SO4，MgCl2·6H2O，K2HPO4And the like.
The strain screening culture medium is an inorganic salt liquid culture medium: na (Na)2HPO4·2H2O，3.5g；K2HPO4，1g；(NH4)2SO4，0.5g；MgCl2·6H2O，0.1g；Ca(NO3)2·4H2O, 0.05 g. Sterilizing at 121 deg.C under high temperature steam for 30 min.
LB liquid/solid medium: 5g of yeast extract; peptone, 10 g; NaCL, 10 g; distilled water, 1000 mL. Adjusting pH to 7.0-7.2, sterilizing at 121 deg.C under high temperature steam for 20min, adding agar 15-20g into solid culture medium based on liquid culture medium, pouring the culture medium into conical flask, and sterilizing at 121 deg.C under high temperature steam for 20 min.
Beef extract solid medium: 3g of beef extract; 5g of NaCL; peptone, 10 g; 15-20g of agar; distilled water, 1000 mL. Adjusting pH to 7.0-7.2, and sterilizing at 121 deg.C under high temperature steam for 20 min.
The fourth concrete implementation mode: the identification of Acinetobacter pitteus (Acinetobacter pitttii) GB-2 of the embodiment:
1. strain morphology identification
Diluting the bacterial suspension with sterile water under aseptic conditions, placing 100 μ L of bacterial liquid on an LB solid medium plate, uniformly coating with a sterilized coating rod, placing the plate upside down in a constant temperature incubator at 37 ℃ for culturing until a single bacterial colony grows out, and observing the characteristics of the bacterial colony, such as shape, size, color, transparency, edge and the like, wherein the morphological diagram of the bacterial colony plate is shown in FIG. 1, and the observation results are shown in Table 1.
TABLE 1 morphological observation of Strain GB-2
2. Physiological and biochemical identification of strain
The strain GB-2 was subjected to physiological and biochemical experiments, and the results were analyzed according to Bergey's Manual of bacteriological identification, and the results are shown in Table 2, wherein "+" indicates positive reaction and "-" indicates negative reaction.
TABLE 2 physiological and biochemical experimental results of strain GB-2
3. Strain 16S rDNA identification
Extracting bacterial DNA, carrying out PCR amplification on the obtained bacterial genome DNA by using bacterial 16S rDNA universal primers, (27F: 5'-AGAGTTTGATCCTGGCTCAG-3', 1492R: 5'-GGTTACCTTGTTACGACTT-3'), wherein the total volume of a PCR amplification system (the total volume is 50 mu L) and reaction conditions are shown in tables 3 and 4, carrying out gel cutting after electrophoresis to recover PCR products, entrusting Cinzhi-Zhi-Biotech limited to carry out 16S rDNA sequencing, and the sequence of the 16S rDNA is shown in SEQ ID NO: 1 is shown.
TABLE 3 PCR amplification reaction System
TABLE 4 PCR amplification conditions
The sequencing result is compared with the 16S rDNA sequence registered in Genbank by using BLAST software, the homology of the strain GB-2 obtained by screening in the embodiment with the Acinetobacter (Acinetobacter pittii) is found to reach 100 percent, and the sequence is submitted to a Genbank database to obtain the registration number of MT 150630. According to the 16S rDNA sequencing result, homologous sequences are found in GenBank databases by using Blast tools provided by NCBI and related software such as MEGA7.0, and a phylogenetic tree is established, as shown in FIG. 2. As can be seen from FIG. 2, the strain has the highest homology with Acinetobacter pitteus, up to 100%, and is finally named Acinetobacter pitteus (Acinetobacter pitttii) GB-2.
The functionality of Acinetobacter pitteus (Acinetobacter pitttii) GB-2 in degrading BDE-47 is verified as follows:
test 1: determination of BDE-47 degrading ability of strain
And adding 100 mu L of BDE-47 n-hexane stock solution of 20mg/L into a 50mL conical flask, and adding 20mL of inorganic salt culture medium after the n-hexane is volatilized so that the final concentration of BDE-47 in the reaction system is 100 mu g/L. Inoculating the strain GB-2 into a degradation system, wherein the inoculation amount is 1%, culturing for 7d in a constant-temperature shaking incubator at 35 ℃ and 150rpm, performing HPLC analysis, determining the residual amount of BDE-47 in a culture solution, and calculating the degradation rate of the strain GB-2 on the BDE-47.
The test shows that the degradation rate of the BDE-47 reaches 70.17 percent when the BDE-47 is cultured for 7 days under the condition that the concentration of the BDE-47 is less than 100 mu g/L. Experiments prove that the BDE-47 degrading bacteria GB-2 have the function of repairing BDE-47 polluted soil, and can provide microbial agent resources for in-situ bioremediation of the BDE-47 polluted soil without adding other chemical agents, so that the BDE-47 degrading bacteria GB-2 have wide application prospects.
Sequence listing
<110> university of Harbin
<120> Acinetobacter pitteus and application thereof
<160> 3
<210> 1
<211> 1278
<212> DNA
<213> Acinetobacter pittii (Acinetobacter pittii)
<400> 1
gcggcggacg ggtgagtaat gcttaggaat ctgcctatta gtgggggaca acatttcgaa 60
aggaatgcta ataccgcata cgtcctacgg gagaaagcag gggatcttcg gaccttgcgc 120
taatagatga gcctaagtcg gattagctag ttggtggggt aaaggcctac caaggcgacg 180
atctgtagcg ggtctgagag gatgatccgc cacactggga ctgagacacg gcccagactc 240
ctacgggagg cagcagtggg gaatattgga caatgggcgc aagcctgatc cagccatgcc 300
gcgtgtgtga agaaggcctt atggttgtaa agcactttaa gcgaggagga ggctactgaa 360
gttaatacct tcagatagtg gacgttactc gcagaataag caccggctaa ctctgtgcca 420
gcagccgcgg taatacagag ggtgcaagcg ttaatcggat ttactgggcg taaagcgcgc 480
gtaggcggct aattaagtca aatgtgaaat ccccgagctt aacttgggaa ttgcattcga 540
tactggttag ctagagtgtg ggagaggatg gtagaattcc aggtgtagcg gtgaaatgcg 600
tagagatctg gaggaatacc gatggcgaag gcagccatct ggcctaacac tgacgctgag 660
gtgcgaaagc atggggagca aacaggatta gataccctgg tagtccatgc cgtaaacgat 720
gtctactagc cgttggggcc tttgaggctt tagtggcgca gctaacgcga taagtagacc 780
gcctggggag tacggtcgca agactaaaac tcaaatgaat tgacgggggc ccgcacaagc 840
ggtggagcat gtggtttaat tcgatgcaac gcgaagaacc ttacctggcc ttgacatagt 900
aagaactttc cagagatgga ttggtgcctt cgggaactta catacaggtg ctgcatggct 960
gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca acccttttcc 1020
ttatttgcca gcgagtaatg tcgggaactt taaggatact gccagtgaca aactggagga 1080
aggcggggac gacgtcaagt catcatggcc cttacggcca gggctacaca cgtgctacaa 1140
tggtcggtac aaagggttgc tacctagcga taggatgcta atctcaaaaa gccgatcgta 1200
gtccggattg gagtctgcaa ctcgactcca tgaagtcgga atcgctagta atcgcggatc 1260
agaatgccgc ggtgaata 1278
<210> 2
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> nucleotide sequence of PCR primer 27F.
<400> 2
AGAGTTTGAT CCTGGCTCAG 20
<210> 3
<211> 19
<212> DNA
<213> Artificial sequence
<220>
<223> PCR primer 1492R nucleotide sequence.
<400> 3
GGTTACCTTG TTACGACTT 19
Claims (2)
1. The Acinetobacter cutaneus is characterized in that the Acinetobacter cutaneus (Acinetobacter pittii) GB-2 is preserved in China center for type culture collection, the preservation address is Wuhan university in Wuhan City, China, the preservation date is 1 month and 19 days 2021, and the preservation number is CCTCC No. M2021107.
2. The use of an acinetobacter cutaneus strain according to claim 1 for degrading environmental pollutants, wherein said environmental pollutants are 2,2 ', 4, 4' -tetrabromobisphenol.
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Cited By (2)
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| CN113046267A (en) * | 2021-03-22 | 2021-06-29 | 哈尔滨师范大学 | Rhodococcus ruber and application thereof |
| CN115058362A (en) * | 2022-06-23 | 2022-09-16 | 哈尔滨师范大学 | Bacterial strain for degrading carbon tetrachloride and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113046267A (en) * | 2021-03-22 | 2021-06-29 | 哈尔滨师范大学 | Rhodococcus ruber and application thereof |
| CN113046267B (en) * | 2021-03-22 | 2022-11-15 | 哈尔滨师范大学 | Rhodococcus ruber and application thereof |
| CN115058362A (en) * | 2022-06-23 | 2022-09-16 | 哈尔滨师范大学 | Bacterial strain for degrading carbon tetrachloride and application thereof |
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| CN113234625B (en) | 2022-05-24 |
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