CN102757907B - Endosulfan degradation stain and application thereof in soil remediation - Google Patents
Endosulfan degradation stain and application thereof in soil remediation Download PDFInfo
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Abstract
The invention discloses an endosulfan degradation stain and application thereof in soil remediation. The endosulfan degradation stain is a strain LD-6 of Stenotrophomonas rhizophila, is preserved in China general microbiological strain preservation administration center On March 2, 2012, and the preservation number is CGMCC No.5839. The endosulfan degradation stain can realize the complete degradation of 100 mg/L of endosulfan within 10 days under an aerobic condition and has low-toxic or non-toxic degradation products and fast environmental degradation; the strain LD-6 is sensitive to common antibiotics, such as tetracycline and kanamycin and tolerant to 13 tested heavy metals and can inhibit the growth of a plurality of phytopathogens; and the endosulfan degradation stain has capability of efficiently degrading the endosulfan in an actual soil environment, can outstandingly shorten the degrading time of the endosulfan and has practical application significance and value.
Description
Technical field
The invention belongs to environmental pollutant biologic treating technique field, be specifically related to bacterium and the application in Soil Environmental Pollution is repaired thereof of a high-efficiency degradation 5a,6,9,9a-hexahydro-6,9-methano-2,4.
Background technology
Agricultural chemicals, as a kind of important capital goods in the agricultural sector, is being brought into play extremely important effect at aspects such as protection agriculture production, raising overall productivity in agriculture, the volume increase of promotion stable food and Practices Sustainable Increasing Income of Farmers.China is pesticide producing big country, is also to use big country, and year usage quantity and average consumption are about respectively 1,200,000 tons and 2.34 kg/ha, rank the first in the world.Yet the utilization ratio of agricultural chemicals is but only 10-20%, rest part all enters soil, water body and atmosphere, has caused serious pollution.According to statistics, the whole nation is subject to the farmland of pesticide residual contamination that ten thousand hectares of 1300-1600 are approximately arranged, and accounts for and can plough 10% of area; The nearly half vegetable pesticide residue exceeds standard, and suffer exit loss nearly 80 hundred million dollar because of the residual problem of exceeding standard of agriculture every year.
China starts to develop organochlorine pesticide from the 50's of 20th century, mainly comprises the products such as phenyl-hexachloride, aldrin, Dieldrin-attapulgite mixture, Niran and heptachlor, and they once brought into play significant role in industrial and agricultural production.But because this compounds has extremely strong inrichment in food chain, be important environmental classes hormone and persistence organic pollutant, major part has stopped producing and using.So far, this pesticide residue is still constantly threatening national health, the benign development of restriction rural economy.
5a,6,9,9a-hexahydro-6,9-methano-2,4 (Endosulfan) is comprised of α and b 5a,6,9,9a-hexahydro-6,9-methano-2,4, commodity match by name red (Thionex), large pellet (Thiodan), be a kind of high-efficiency broad spectrum organochlorine insecticide, relative low price, be widely used in the pest control of the crops such as tealeaves, cotton, fruit tree and vegetables.5a,6,9,9a-hexahydro-6,9-methano-2,4 belongs to riskiest pesticide, poisonous to animal especially fish, can damage central nervous system, immunity system, the reproductive system of animal, and the organs such as brain, liver and kidney.The 5a,6,9,9a-hexahydro-6,9-methano-2,4 residual period, is long, is one of several verieties that in organochlorine insecticide, Environmental Residues concentration is the highest, has bioconcentration, and the transformation period in soil reaches 6-11 month.
At present, by technology such as artificial enrichment culture, many bacterium and fungies with degraded organochlorine pesticide compound ability have been isolated.Wherein, bacterium is adaptable owing to having, breeding is fast and the characteristics such as quick of degrading occupy main status.Yet, the efficient degrading bacteria of the tool 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation capability be separated to is still very limited, most bacterial strains can't be realized degradable to 100 mg/L 5a,6,9,9a-hexahydro-6,9-methano-2,4s in 10 days, and degraded product mostly be toxicity more greatly, residual period is longer and environmental hazard is larger 5a,6,9,9a-hexahydro-6,9-methano-2,4 vitriol (Endosulf sulfate).The people such as Li Wen once reported the efficient pale bacillus (C7) of a strain tool 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation capability, but this bacterial strain to the degradation capability of 50 mg/L 5a,6,9,9a-hexahydro-6,9-methano-2,4s in 90.2% left and right, can't realize degradable (degradation rate is mg/L every days 5.64) (Li Wen, Peng Xiang, Zhang Jingshun, Zhang Chen, Yan Yanchun.The screening of 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria and degradation property research.The Shandong agricultural sciences, 2009,1:67-70).Reported that the bacterial strain that degradation rate is the highest is viride (Trichoderma viride) H082, this bacterial strain can be in 7 days efficient degradation 100 mg/L 5a,6,9,9a-hexahydro-6,9-methano-2,4s, it is red that degradation rate reaches 95%(Zhu Li, Wu Jian, Sun Dongchang, Shi Yuefeng.The separation of 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria H082, screening and bacteriostasis property research), but can't realize degradable.In addition, bacterial strain H082 belongs to filamentous fungus, with bacterial isolates, compares, and exist following obviously not enough: 1) growth velocity is slow.The bacterium industrial fermentation is generally between 2-3 days, and the filamentous fungus fermentation is no less than 4, how between 6-8 day.With regard to the general industry fermentation, the ferment product cell concentration can reach 1-5 * 10
9cFU/mL(CFU, colony-forming unit), fungus solids fermentation spore count will hang down an order of magnitude; 2) the fermentation equipment versatility is poor.Filamentous fungus fermentation solid fermentation bed or the fermentor tanks of adopting, equipment interoperability is poor more.Fermentation using bacteria also can be used for secondary metabolite production with fermentor tank except producing thalline, and as microbiotic, pigment and VITAMIN etc., universal performance is good; 3) the microbial inoculum application effect is relatively slow.After the bacterium microbial inoculum sprays, bacterium directly acts on pesticide residue, and the filamentous fungus spore needs can act on after sprouting, forming mycelia.In addition, 5a,6,9,9a-hexahydro-6,9-methano-2,4 meta-bolites, antibiotics resistance and the heavy metal tolerance etc. of above-mentioned two strain degradation bacteria are all not yet verified, and exist certain environment to use risk.In sum, screening is efficient, the tolerance heavy metal, to antibiotic sensitive, and the 5a,6,9,9a-hexahydro-6,9-methano-2,4 bacterium for degrading of the non-5a,6,9,9a-hexahydro-6,9-methano-2,4 vitriol of degraded product becomes study hotspot.
Summary of the invention
What the objective of the invention is for above-mentioned existing 5a,6,9,9a-hexahydro-6,9-methano-2,4 microorganism, 5a,6,9,9a-hexahydro-6,9-methano-2,4 degraded to be existed can't realize degradable to 100 mg/L 5a,6,9,9a-hexahydro-6,9-methano-2,4s in 10 days, and degraded product mostly is the defect of the 5a,6,9,9a-hexahydro-6,9-methano-2,4 vitriol (Endosulf sulfate) that toxicity is larger, residual period is longer and environmental hazard is larger, and a kind of efficient 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria is provided.
Another object of the present invention is to provide the application of above-mentioned bacterial classification in repairing the 5a,6,9,9a-hexahydro-6,9-methano-2,4 contaminated soil.
Above-mentioned technical problem of the present invention is implemented by the following technical programs:
One strain 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria, this bacterium is the bacterial strain LD-6 that has a liking for root Stenotrophomonas (Stenotrophomonas rhizophila), on March 2nd, 2012 in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, be numbered CGMCC No.5839, this bacterium has the ability of efficient degradation 5a,6,9,9a-hexahydro-6,9-methano-2,4.The address of depositary institution is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, and the Classification And Nomenclature of this bacterium is for having a liking for root Stenotrophomonas Stenotrophomonas rhizophila.
The application of a kind of described 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria in being subject to the reparation of 5a,6,9,9a-hexahydro-6,9-methano-2,4 contaminated soil.
As preferably, described bacterial strain CGMCC No.5839 is seeded in the basic inorganic salt liquid substratum containing 5a,6,9,9a-hexahydro-6,9-methano-2,4 50-100 mg/L, more than 48h is cultivated in 120 r/min concussions, described basic inorganic salt liquid Media Components is (g/L): MgCl
20.2, NH
4nO
31, KH
2pO
42, K
2hPO
47.5 NaCl 1,5a,6,9,9a-hexahydro-6,9-methano-2,4 0.1, deionized water 1 L.
As preferably, described liquid culture temperature is 25-30 ℃, and the pH of basic inorganic salt liquid substratum is 5.4-6.8.
As preferably, described liquid culture temperature is 30 ℃.
As preferably, described bacterial strain CGMCC No.5839 is seeded to 30 ℃ of constant temperature culture in fermention medium and, more than 24 hours, obtains the thalline fermented liquid, to being subject to the 5a,6,9,9a-hexahydro-6,9-methano-2,4 contaminated soil to spray the thalline fermented liquid, degraded; The composition of fermention medium is (g/L): Semen Maydis powder 10 g/L, bean cake powder 8 g/L, (NH
4)
2sO
46 g/L, corn steep liquor 6 g/L, MgSO
47H
2o 1 g/L, K
2hPO
43H
2o 2 g/L, MnSO
40.05 g/L, pH 7.2.
5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria provided by the present invention
stenotrophomonas rhizophilalD-6 is derived near the mud of Tonglu, Zhejiang insecticide factory sewage draining exit, through artificial enrichment culture, pressure screening and separation and purification, obtains.This Pseudomonas is had a liking for the root Stenotrophomonas, and its Phylogenetic analysis as shown in Figure 1.Its physiological and biochemical property is: Gram-negative, the catalase positive, oxidase negative, V. P. negative, the indole test feminine gender, obligate is supported well, can be 4 ℃ of growths, but can't survive in 40 ℃, can take wood sugar as carbon source, thalli morphology is rod-short, that bacterium colony is is light yellow, circular, edge is neat, smooth moistening, the GenBank accession number of bacterial strain 16S rDNA is JQ670922, on March 2nd, 2012, in the preservation of Chinese common micro-organisms culture presevation administrative center, is numbered CGMCC No.5839.
The optimum growing condition of this bacterium is: pH 7.2, and temperature is 30 ℃.With 1/5th LB(Luria-Bertani) or industrial fermentation substratum (Semen Maydis powder 10 g/L, bean cake powder 8 g/L, (NH
4)
2sO
46 g/L, corn steep liquor 6 g/L, MgSO
47H
2o 1 g/L, K
2hPO
43H
2o 2 g/L, MnSO
40.05 g/L, pH 7.2) to cultivate after 24-36 hour, bacterial concentration can reach 5-8 * 10
9cFU/mL.
This bacterium is to tsiklomitsin and kantlex sensitivity, and the two is all common antibiotics.Can think: when thalline is released in physical environment, can does not produce superbacteria or propagate between indigenous bacterium because of anti-(anti-) property of medicine problem.
This bacterium to Candida albicans (
candida albicans), dry thread Pyrenomycetes (
rhizoctonia solani), sclerotinite (
sclerotinia sclerotiorum), verticillium wilt pathogen (
verticillium dahliae), fusarium graminearum (
fusarium graminearum) and the capsicum wilt bacterium (
fusarium oxysporum) present obvious restraining effect.Can think: after thalline is used, can play to a certain extent the effect that suppresses or alleviate the Plant diseases generation.
This bacterium is to 13 kinds of test metal ion (Li
+, Ag
+, Cu
+, Cu
2+, Hg
2+, Ba
2+, Pb
2+, Zn
2+, Mn
2+, Ni
2+, Fe
2+, Fe
3+and Co4
+) present tolerance in various degree.In view of China's Pollution In Soil---persistence organic pollutant pollution and heavy metal contamination are also deposited, and the pesticide residual degradation bacterial strain possesses heavy metal tolerance can be very important.Can think: under the general soil envrionment conditions, this thalline is used repairing effect and is subject to the heavy metal inhibitory effect less.
This bacterial strain is the (triangular flask that volume is 250 mL in the shaking flask Degrading experiment, 100 mL liquid minimal mediums are housed, 5a,6,9,9a-hexahydro-6,9-methano-2,4 concentration is 100 mg/L, shaking speed is 160 r/min, temperature is 30 ℃), can be by 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradable (degradation rate be mg/L every days 10) in 10 days, meta-bolites is 5a,6,9,9a-hexahydro-6,9-methano-2,4 ether and 5a,6,9,9a-hexahydro-6,9-methano-2,4 glycol, the two toxicity is much smaller than 5a,6,9,9a-hexahydro-6,9-methano-2,4, and environment degradable is quick.Be in 50 mg/kg situations at soil 5a,6,9,9a-hexahydro-6,9-methano-2,4 content, use this bacterium and within 28 days, can make to degrade for 68.6% 5a,6,9,9a-hexahydro-6,9-methano-2,4 in the examination soil sample, significantly promote the 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation rate, shorten the duration of degrading, possess the practical application meaning and value.
This bacterium is to have reported in degradation bacteria, and what a unique strain possessed the 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation capability has a liking for the root Stenotrophomonas.
In sum, compared to the prior art the present invention has following advantage:
Compared with prior art, of the present invention
stenotrophomonas rhizophilalD-6 be the first strain possess the 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation capability have a liking for the root Stenotrophomonas, the 5a,6,9,9a-hexahydro-6,9-methano-2,4 of can usining under aerobic condition carries out growth and breeding as sole carbon source and sulphur source, simultaneously by its fast degradation.Under the pure culture condition, the 5a,6,9,9a-hexahydro-6,9-methano-2,4 that it can be 100 mg/L by concentration was degraded fully in 10 days.With existing 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria strains, compare; this bacterium is when possessing higher 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation rate (being only second to above-mentioned viride H082); to the common antibiotics sensitivity; can tolerate 13 heavy metal species; the various plants pathogenic micro-organism is existed to restraining effect; degraded product low toxicity or nontoxic; the industrial scale fermentative production is convenient, fermented liquid cell concentration high (being better than viride H082); and possess the ability of performance 5a,6,9,9a-hexahydro-6,9-methano-2,4 Degradation under actual edatope condition, this bacterial strain is expected to play a significant role in the coenocorrelation reparation of 5a,6,9,9a-hexahydro-6,9-methano-2,4 contaminated soil.
The accompanying drawing explanation
The Phylogenetic analysis of Fig. 1, bacterial strain LD-6.
The degradation capability test of Fig. 2, bacterial strain LD-6, wherein ■: α 5a,6,9,9a-hexahydro-6,9-methano-2,4; ▲: the b 5a,6,9,9a-hexahydro-6,9-methano-2,4; ◆: α and b 5a,6,9,9a-hexahydro-6,9-methano-2,4 sum.
5a,6,9,9a-hexahydro-6,9-methano-2,4 degraded situation, wherein in Fig. 3, bacterial strain LD-6 edatope: α 5a,6,9,9a-hexahydro-6,9-methano-2,4 content (contrast) in the deactivation soil sample; ■: spray α 5a,6,9,9a-hexahydro-6,9-methano-2,4 content in LD-6 thalline deactivation soil sample; △: α 5a,6,9,9a-hexahydro-6,9-methano-2,4 content (contrast) in fresh soil sample; ▲: spray α 5a,6,9,9a-hexahydro-6,9-methano-2,4 content in LD-6 thalline fresh soil sample; ◇: b 5a,6,9,9a-hexahydro-6,9-methano-2,4 content (contrast) in the deactivation soil sample; ◆: spray b 5a,6,9,9a-hexahydro-6,9-methano-2,4 content in LD-6 thalline deactivation soil sample; Zero: b 5a,6,9,9a-hexahydro-6,9-methano-2,4 content (contrast) in fresh soil sample; ●: spray b 5a,6,9,9a-hexahydro-6,9-methano-2,4 content in LD-6 thalline fresh soil sample.
Embodiment
Below by specific embodiment, technical scheme of the present invention is described in further detail.Should be appreciated that enforcement of the present invention is not limited to the following examples, any pro forma accommodation that the present invention is made and/or change all will fall into protection domain of the present invention.
In the present invention, if not refer in particular to, all part, per-cents are weight unit, and all equipment and raw material etc. all can be buied from market or the industry is commonly used.
embodiment 1: stenotrophomonas rhizophilathe separation of LD-6 and 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation capability
Near mud collection in worksite Tonglu, Zhejiang insecticide factory sewage draining exit, with 150 mL triangular flask splendid attires, and take back rapidly laboratory.The sample of fetching is placed in to the inorganic salt liquid substratum domestication of 5a,6,9,9a-hexahydro-6,9-methano-2,4 selectivity basis to be cultivated.Media Components is (g/L): MgCl
20.2, NH
4nO
31, KH
2pO
42, K
2hPO
47.5 NaCl 1,5a,6,9,9a-hexahydro-6,9-methano-2,4 0.1, and deionized water 1 L, 6.8,115 ℃ of moist heat sterilizations of pH are standby after 30 minutes.Culture condition is: 28 ℃, 120 r/min concussions are cultivated, and the container liquid amount is vessel volume 1/3.After this, every 6-7 days, by shaking flask static a moment, abandon supernatant liquor, fill into subsequently equivalent fresh configuration the selected liq substratum and continue the concussion enrichment culture.After surrounding, above-mentioned culture is evenly coated on solid selectivity basis minimal medium (agar that adds again 20 grams in every liter of liquid nutrient medium).Coated flat-plate inverted postpone is put into to incubator, cultivate 72 hours for 28 ℃.With aseptic toothpick select single bacterium colony or with inoculating needle line until occur single bacterium colony again row be forwarded on the solid selectivity flat board of fresh configuration, and verify one by one that by vapor-phase chromatography the 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation capability of each bacterial strain, result obtain a strain 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria LD-6.
With OD
600the inoculum size that value is 0.5 be take bacterial strain LD-6 access in the selectivity basis minimal medium that 100 mg/L 5a,6,9,9a-hexahydro-6,9-methano-2,4s are sole carbon source and sulphur source, and 30 ℃, 160 r/min concussions are cultivated.Subsequently, every 2 days to substratum in the residual condition of 5a,6,9,9a-hexahydro-6,9-methano-2,4 tested, result is as shown in Figure 2.Therefrom can find out that bacterial strain LD-6 can 5a,6,9,9a-hexahydro-6,9-methano-2,4 be to be grown in sole carbon source and sulphur source, realize degradable to 5a,6,9,9a-hexahydro-6,9-methano-2,4 simultaneously.
The present embodiment explanation separation obtains
stenotrophomonas rhizophilalD-6 can utilize 5a,6,9,9a-hexahydro-6,9-methano-2,4 to carry out growth and breeding as sole carbon source and sulphur source, and has the ability of efficient degradation 5a,6,9,9a-hexahydro-6,9-methano-2,4.
embodiment 2: stenotrophomonas rhizophilathe resistance experiment of LD-6
Adopt the filter paper method, select tsiklomitsin and kantlex to be tested, the antibacterial circle diameter size with each microbiotic filter paper (30 μ g/ sheet) on cultivation degradation bacteria LD-6 flat board, as the criterion of sensitivity or resistance.Test-results shows, antibacterial circle diameter is respectively 24 and 16 mm, and (the medium sensitivity scope is: tsiklomitsin 23-25 mm to be the medium sensitivity reaction; Kantlex 14-22 mm).
The present embodiment explanation, can be because anti-(anti-) property of medicine problem produces superbacteria or propagates between indigenous bacterium, for its practical application from now on provides safety assurance when thalline is released in physical environment.
embodiment 3: stenotrophomonas rhizophilaheavy metal minimum inhibition concentration (MIC) test of LD-6
The MIC Test Design is carried out (Filali B.K. with reference to the people's such as Filali research, Taoufik J., Zeroual Y., Dzairi F.Z., Talbi M., Blaghen M. Waste water bacterial isolates resistant to heavy metals and antibiotics. Current Microbiology, 2000,41 (3): 151-156), picking etc. are large, the mono-bacterium colony of the LD-6 of activated processing is in culture test tube, 30 ℃, 160 rpm shaking culture 48 hours.Judge MICs according to the thalline growing way, establish 3 repetitions for every group, test respectively the tolerance situation of this bacterium to 13 metal ion species.Result is as shown in table 1, the multiple tolerance of this bacterial strain tool, and tolerance is strong.
embodiment 4: stenotrophomonas rhizophilathe phytopathogen inhibition test of LD-6
Method (Zhu Lihong, Wu Jian, Sun Dongchang, Shi Yuefeng with reference to the people such as Zhu Lihong report.The separation of 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria H082, screening and bacteriostasis property research) carry out the bacteriostasis test of bacterial strain LD-6.Result is as shown in table 2, and this bacterial strain presents restraining effect to multiple for the examination pathogenic bacteria.The present embodiment explanation thalline can play the effect that suppresses or alleviate the Plant diseases generation after using to a certain extent.
annotate:+ bacteriostasis rate 50-80%; ++ bacteriostasis rate is more than 80%.
embodiment 5: stenotrophomonas rhizophilathe test of 5a,6,9,9a-hexahydro-6,9-methano-2,4 in LD-6 degraded soil
Bacterial strain LD-6 30 ℃ of constant temperature culture in fermention medium, after 24 hours, are obtained to the thalline fermented liquid.The composition of fermention medium is (g/L): Semen Maydis powder 10 g/L, bean cake powder 8 g/L, (NH
4)
2sO
46 g/L, corn steep liquor 6 g/L, MgSO
47H
2o 1 g/L, K
2hPO
43H
2o 2 g/L, MnSO
40.05 g/L, pH 7.2.
Add a certain amount of 5a,6,9,9a-hexahydro-6,9-methano-2,4 (5a,6,9,9a-hexahydro-6,9-methano-2,4 is dissolved among acetone in advance) in sterilized water, after on shaking table, fully concussion mixes, soak soil to be tried (moisture soil that is pH 5.4 for examination soil), make soil particle can adsorb uniformly 5a,6,9,9a-hexahydro-6,9-methano-2,4,5a,6,9,9a-hexahydro-6,9-methano-2,4 content is 50 mg/kg.Spray the thalline fermented liquid, stir, make LD-6 cell concentration in soil sample be about 1 * 10
8individual/g.Soil water content is controlled at 60% left and right, 25 ℃ of constant temperature.Simultaneously, with reference to the people such as Li Wen research (Li W., Dai Y., Xue B.B., Li Y.Y., Peng X., Zhang J.S., Yan Y.C. Biodegradation and detoxification of endosulfan in aqueous medium and soil by
achromobacter xylosoxidansstrain CS5. Journal of Hazardous Materials, 2009,167:209-216), contrast is set.Subsequently, got 5 g soil sample measuring and calculating 5a,6,9,9a-hexahydro-6,9-methano-2,4 degraded situations every 7 days.
Result as shown in Figure 3, is degraded for the 5a,6,9,9a-hexahydro-6,9-methano-2,4 that has 68.6% in the examination soil sample in 28 days, sprays LD-6 and can significantly promote the 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation rate, shorten the degraded duration, possesses the practical application meaning and value.
Claims (6)
1. a strain 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria, this bacterium be have a liking for the root Stenotrophomonas (
stenotrophomonas rhizophila) bacterial strain LD-6, in the preservation of Chinese common micro-organisms culture presevation administrative center, be numbered CGMCC No.5839 on March 2nd, 2012, this bacterium has the ability of degraded 5a,6,9,9a-hexahydro-6,9-methano-2,4.
2. the application of the described 5a,6,9,9a-hexahydro-6,9-methano-2,4 degradation bacteria of claim 1 in being subject to the reparation of 5a,6,9,9a-hexahydro-6,9-methano-2,4 contaminated soil.
3. application claimed in claim 2, it is characterized in that: described bacterial strain CGMCC No.5839 is seeded to the basic inorganic salt liquid substratum for liquid culture containing 5a,6,9,9a-hexahydro-6,9-methano-2,4 50-100 mg/L, more than 48 h are cultivated in 120 r/min concussions, described basic inorganic salt liquid Media Components is: MgCl
20.2 g/L, NH
4nO
31 g/L, KH
2pO
42 g/L, K
2hPO
47.5 g/L, NaCl 1 g/L, 5a,6,9,9a-hexahydro-6,9-methano-2,4 0.1 g/L, deionized water 1 L.
4. application claimed in claim 3 is characterized in that: described liquid culture temperature is 25-30 ℃, and the pH of basic inorganic salt liquid substratum is 5.4-6.8.
5. application claimed in claim 3 is characterized in that: described liquid culture temperature is 30 ℃.
6. application claimed in claim 2 is characterized in that: described bacterial strain CGMCC No.5839 is seeded to 30 ℃ of constant temperature culture in fermention medium and, more than 24 hours, obtains the thalline fermented liquid, degraded to being subject to the 5a,6,9,9a-hexahydro-6,9-methano-2,4 contaminated soil to spray the thalline fermented liquid; The composition of fermention medium is: Semen Maydis powder 10 g/L, bean cake powder 8 g/L, (NH
4)
2sO
46 g/L, corn steep liquor 6 g/L, MgSO
47H
2o 1 g/L, K
2hPO
43H
2o 2 g/L, MnSO
40.05 g/L, pH 7.2.
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| CN103060233B (en) * | 2012-12-27 | 2014-07-23 | 山东农业大学 | A Bordetella petrii NS and applications of an immobilized enzyme thereof in the soil |
| CN105624063A (en) * | 2016-02-04 | 2016-06-01 | 云南圣清环保科技有限公司 | Efficient mixed flora for treating sludge |
| CN106434497B (en) * | 2016-12-12 | 2019-06-04 | 浙江农林大学 | Siderophore high yield bacteria preparation and its application in terms of Heavy Metals in Soil Contaminated reparation |
| CN115094013B (en) * | 2022-08-25 | 2022-11-22 | 江苏聚庚科技股份有限公司 | Stenotrophomonas rhizophila, microbial inoculum and application of stenotrophomonas rhizophila in wastewater treatment |
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| ID:JQ6709221;Yu F.B ET AL;《GenBank》;20120327;1-2 * |
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