CN105483046A - Cyanobacteria culture medium - Google Patents

Cyanobacteria culture medium Download PDF

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Publication number
CN105483046A
CN105483046A CN201511023126.1A CN201511023126A CN105483046A CN 105483046 A CN105483046 A CN 105483046A CN 201511023126 A CN201511023126 A CN 201511023126A CN 105483046 A CN105483046 A CN 105483046A
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China
Prior art keywords
substratum
blue
green algae
culture medium
cyanobacteria
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CN201511023126.1A
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马奎
王铜山
石书缘
赵伟波
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China Petroleum and Natural Gas Co Ltd
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China Petroleum and Natural Gas Co Ltd
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Priority to CN201511023126.1A priority Critical patent/CN105483046A/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The invention provides a cyanobacteria culture medium. The cyanobacteria culture medium is prepared from a Bg11 culture medium and triuranium octoxide, wherein the mass concentration of the triuranium octoxide is 1 ppm-100 ppm when the total mass of the Bg11 culture medium is taken as the criterion. The invention further provides a method for verifying the influence of radioactive substances on cyanobacteria culture. The method comprises the steps that the triuranium octoxide is added into the Bg11 culture medium, and a test for the growth speed and organic matter enrichment is performed on obtained cyanobacteria, wherein the dosage of the triuranium octoxide is 1 ppm-100 ppm when the total mass of the Bg11 culture medium is taken as the criterion. The cyanobacteria culture medium can enable the cyanobacteria to be prosperous and enable cyanobacteria organic matter to be extensively enriched.

Description

A kind of blue-green algae substratum
Technical field
The present invention relates to a kind of substratum, particularly a kind of blue-green algae substratum, belong to ancient oil-gas exploration experimental technique field.
Background technology
Blue-green algae is as the main source material of Neoproterozoic hydrocarbon source rock in North China, and its developmental potentiality and scale are to the ancient oil-gas generation important of Singapore dollar in North China.Probe into the geologic agent affecting blue-green algae and grow, developmental potentiality and the Scale Revenue Ratio of centering Neoproterozoic blue-green algae are significant.Qie Erruo Baily and Fukushima nuclear incident zone of pollution make the extraordinary growth of the biology of Adjacent Sea, the normal gamma ray curve of Ordos Basin XX well (containing radioactivity) and organic carbon curve have good anastomose property, and the volcanics section of concentrating (high radioactivity band) usually grows high quality source rock etc., this series of embodiment implies radioactive substance and is conducive to biological super misgrowth and has direct or indirect contact.
The domestic research of the growing state about blue-green algae under radioactivity is at present in the blank stage, and more is the impact of growing blue-green algae in research radiativity condition, and the conclusion drawn is that the growth of radiation to blue-green algae has destruction.And radiation and radiation have essential distinction, what kind of impact radioactive material confrontation blue-green algae has also unknown.
Summary of the invention
In order to solve the problems of the technologies described above, the object of the present invention is to provide a kind of blue-green algae substratum, this blue-green algae substratum can impel blue-green algae to grow prosperity, and impels blue-green algae organic matter that a large amount of enrichment occurs.
In order to realize above-mentioned technical purpose, the invention provides a kind of blue-green algae substratum, this blue-green algae substratum comprises Bg11 substratum and uranous uranic oxide, and wherein, with the total mass of Bg11 substratum for benchmark, the mass concentration of uranous uranic oxide is 1ppm-100ppm.
In above-mentioned blue-green algae substratum provided by the invention, preferably with the total mass of Bg11 substratum for benchmark, the mass concentration of uranous uranic oxide is 3ppm, 15ppm and 75ppm.
According to the specific embodiment of the present invention, the uranous uranic oxide of employing extract from rock asphalt uranium ore.
In above-mentioned blue-green algae substratum provided by the invention, preferably, the pH value of the Bg11 substratum of employing is 5-9.
In above-mentioned blue-green algae substratum provided by the invention, preferably, the pH value of the Bg11 substratum of employing is 6-8.
In above-mentioned blue-green algae substratum provided by the invention, preferably, intensity of illumination when this blue-green algae substratum is specifically for cultivating blue-green algae is 10 μm of ol/m 2/ s-50 μm of ol/m 2/ s.
In above-mentioned blue-green algae substratum provided by the invention, preferably, intensity of illumination when this blue-green algae substratum is specifically for cultivating blue-green algae is 20 μm of ol/m 2/ s-30 μm of ol/m 2/ s.
In above-mentioned blue-green algae substratum provided by the invention, preferably, envrionment temperature when this blue-green algae substratum is specifically for cultivating blue-green algae is 10 DEG C-40 DEG C.
In above-mentioned blue-green algae substratum provided by the invention, preferably, envrionment temperature when this blue-green algae substratum is specifically for cultivating blue-green algae is 25 DEG C-27 DEG C.
Present invention also offers a kind of method verifying the impact that radioactive material confrontation blue-green algae is cultivated, the method comprises the following steps:
In Bg11 substratum, add uranous uranic oxide, to the blue-green algae obtained carry out the speed of growth and and the test of Organic Matter Enrichment, wherein, with the total mass of Bg11 substratum for benchmark, the addition of described uranous uranic oxide is 1ppm-100ppm.
In the method for the impact that above-mentioned checking radioactive material confrontation blue-green algae provided by the invention is cultivated, preferably, with the total mass of Bg11 substratum for benchmark, the addition of uranous uranic oxide is 3ppm, 15ppm and 75ppm.
In the method for the impact that above-mentioned checking radioactive material confrontation blue-green algae provided by the invention is cultivated, when cultivating blue-green algae, the pH value of the Bg11 substratum of employing is 5-9, and preferably, the pH value of the Bg11 substratum of employing is 6-8; Intensity of illumination is 10 μm of ol/m 2/ s-50 μm of ol/m 2/ s, preferably, intensity of illumination is 20 μm of ol/m 2/ s-30 μm of ol/m 2/ s; Envrionment temperature is 10 DEG C-40 DEG C, and preferably, envrionment temperature is 25 DEG C-27 DEG C;
Above-mentioned blue-green algae substratum provided by the invention can make blue-green algae oil quantity while normal proliferative increase substantially, for research radioactive substance impels the low microfossil Organic Matter Enrichment that waits to provide a kind of research method.
Accompanying drawing explanation
Fig. 1 is the electron micrograph sheet that reference examples 1 adopts corresponding culture medium culturing Microcystis aeruginosa gained;
Fig. 2 is the electron micrograph sheet that embodiment 1 adopts corresponding culture medium culturing Microcystis aeruginosa gained;
Fig. 3 is the electron micrograph sheet that embodiment 2 adopts corresponding culture medium culturing Microcystis aeruginosa gained;
Fig. 4 is the electron micrograph sheet that embodiment 3 adopts corresponding culture medium culturing Microcystis aeruginosa gained;
Fig. 5 is the electron micrograph sheet that reference examples 2 adopts corresponding culture medium culturing anabena gained;
Fig. 6 is the electron micrograph sheet that embodiment 4 adopts corresponding culture medium culturing anabena gained;
Fig. 7 is the electron micrograph sheet that embodiment 5 adopts corresponding culture medium culturing anabena gained;
Fig. 8 is the electron micrograph sheet that embodiment 6 adopts corresponding culture medium culturing anabena gained;
Fig. 9 is that reference examples 1 adopts micro-algae change of production figure every day of corresponding culture medium culturing Microcystis aeruginosa gained to embodiment 1-3;
Figure 10 is that reference examples 2 adopts micro-algae change of production figure every day of corresponding culture medium culturing anabena gained to embodiment 4-6.
Embodiment
In order to there be understanding clearly to technical characteristic of the present invention, object and beneficial effect, existing following detailed description is carried out to technical scheme of the present invention, but can not be interpreted as to of the present invention can the restriction of practical range.
Reference examples is set respectively and embodiment is described blue-green algae substratum of the present invention.By often organizing before reference examples and embodiment cultivate micro-algae, the process obtaining algae kind can also being comprised, by adopting a large amount of algae kind of quick obtaining over full culture medium culturing 6-10 days, also can obtain algae kind by commercial means.
Following often organize reference examples and embodiment except adopted substratum and cultural method different except, all the other parameters of the algae kind that all the other adopt, culture device, external environment and substratum are as all identical in pH value etc.
Respectively two groups of reference examples and two groups of embodiments and experimental example are described.
The microcystic aeruginosa kind cultivated in the Anabaena Flos-aquae kind of cultivating in reference examples 1 and embodiment 1 and reference examples 2 and embodiment 2, all puts into the trilateral glass dish of 250mL.After inoculation, initial copper Anabaena Flos-aquae liquid OD value is about 0.025, and microcystic aeruginosa liquid OD value is about 0.07.Culture temperature controls at 25 DEG C-27 DEG C.There is provided light source by fluorescent lamp, Light To Dark Ratio example is 12h:12h, and intensity of illumination is 20 μm of ol/m 2/ s-30 μm of ol/m 2/ s, pH value control 7-8.The cultivation time is 12 days.
The cultivation of blue-green algae kind: mainly by a large amount of algae kind of quick obtaining over culture medium culturing 7-10 days.Also algae kind can be obtained by commercial means.
Radioactive mineral: uranous uranic oxide pressed powder, for extracting in rock asphalt uranium ore, is obtained by commercial means.
Method for testing and analyzing:
Growth indexes measures: every 1 day after inoculation, get different parallel group of algae liquid 3mL respectively and detect OD680 in Shimadzu UV-2000 spectrophotometer
Micro-and the transmission electron microscope observing of algae kind: the algae kind of different treatment of learning from else's experience is observed under OlympusBX51 microscope and projection Electronic Speculum
Grease extracts and assay:
Prepare a 5mL size, with the vial of tetrafluoroethylene lid, accurately take lyophilize algae powder M1mg, the algae powder claimed is transferred in above-mentioned vial, and add magnetic stir bar, add the dimethyl sulfoxide (DMSO) methanol solution (V/V) of 10%, extracting 30 minutes is stirred at 50 DEG C of water bath condition lower magnetic forces, with after through 4000rpm centrifugal 10 minutes, supernatant liquor is transferred in another 25mL vial, extract twice in this way, ether is added: normal hexane mixed solution (1:1V/V) in residue algae-residue, magnetic agitation drawer 30 minutes, 4000rpm is centrifugal afterwards, supernatant liquor is transferred in above-mentioned 25ml vial, extract 3 times in this way, pure water is added in the supernatant liquor of 5 extractions, make normal hexane, ether, methyl alcohol, water four kinds of ratios are 1:1:1:1, through the centrifugal 5min of 1500rpm after abundant concussion mixing, pipette upper organic phase, lower floor's inorganic phase adds ether again: normal hexane mixed solution (1:1), centrifugal extraction twice, prepare the vial of a 1.5ml capacity and the M2 that weighs, after upper organic phase merges after nitrogen is concentrated, transfer concentrated solution is to above-mentioned vial, nitrogen is concentrated into lyophilize again after constant weight, weigh, total lipid content LC (%DW) is calculated as follows: %Totallipids=100 × [M3-M2]/M1.
Reference examples 1
What this reference examples adopted is the Bg11 substratum of the 100mL of standard;
Anabaena Flos-aquae kind is inoculated into described Bg11 substratum, and arranges 3 parallel group.
The Anabaena Flos-aquae kind Bg11 substratum prepared is diluted, is inoculated in the trilateral glass cylinder container of 250mL.
Embodiment 1
What the present embodiment adopted is the Bg11 substratum of the 100mL of standard, and adds the uranous uranic oxide of 3ppm mass concentration in the medium respectively;
Add substratum described in microcystic aeruginosa kind being inoculated into dilute, the substratum with the uranous uranic oxide of this mass concentration arranges 3 parallel group.
Embodiment 2
What the present embodiment adopted is the Bg11 substratum of the 100mL of standard, and adds the uranous uranic oxide of 15ppm mass concentration in the medium respectively;
Add substratum described in microcystic aeruginosa kind being inoculated into dilute, the substratum with the uranous uranic oxide of this mass concentration arranges 3 parallel group.
Embodiment 3
What the present embodiment adopted is the Bg11 substratum of the 100mL of standard, and adds the uranous uranic oxide of 75ppm mass concentration in the medium respectively;
Add substratum described in microcystic aeruginosa kind being inoculated into dilute, the substratum with the uranous uranic oxide of this mass concentration arranges 3 parallel group.
Reference examples 2
What this reference examples adopted is the Bg11 substratum of the 100mL of standard;
Anabena kind is inoculated into Bg11 substratum, and arranges 3 parallel group;
The Anabaena Flos-aquae kind Bg11 substratum prepared is diluted, is inoculated in the trilateral glass cylinder container of 250mL.
Embodiment 4
What the present embodiment adopted is the Bg11 substratum of the 100mL of standard, and adds the uranous uranic oxide of 3ppm mass concentration in the medium respectively;
Add substratum described in anabena kind being inoculated into dilute, the substratum with the uranous uranic oxide of this mass concentration arranges 3 parallel group.
Embodiment 5
What the present embodiment adopted is the Bg11 substratum of the 100mL of standard, and adds the uranous uranic oxide of 15ppm mass concentration in the medium respectively;
Add substratum described in anabena kind being inoculated into dilute, the substratum with the uranous uranic oxide of this mass concentration arranges 3 parallel group.
Embodiment 6
What the present embodiment adopted is the Bg11 substratum of the 100mL of standard, and adds the uranous uranic oxide of 75ppm mass concentration in the medium respectively;
Add substratum described in anabena kind being inoculated into dilute, the substratum with the uranous uranic oxide of this mass concentration arranges 3 parallel group.
Experimental result
See Fig. 1, Fig. 2, Fig. 3 and Fig. 4, cell microscopic photographic result also shows, under uranous uranic oxide material radioactivity, the cellular form of Microcystis aeruginosa, without considerable change, does not morph;
See Fig. 9, with reference to reference examples 1, in embodiment 1-3, the increment of Microcystis aeruginosa is relatively many, and the Microcystis aeruginosa promoter action in embodiment 1 is the most obvious, embodiment 2 and embodiment 3 promoter action relatively little;
See table 1, for being the total lipid content of capsule algae after cultivation in reference examples 1 and embodiment 1-3, the fat content of relative comparison example 1, the fat content of embodiment 1 and embodiment 2 increases on a small quantity, and the fat content of embodiment 3 increases considerably, and is about three times of reference examples.
The different uranium ore material concentration of table 1 is on the impact of Microcystis aeruginosa fat content
Numbering Reference examples 1 Embodiment 1 Embodiment 2 Embodiment 3
Fat content (%) 9.09 10.4 9.55 31.52
See Fig. 5, Fig. 6, Fig. 7 and Fig. 8, cell microscopic photographic result also shows, under uranous uranic oxide material radioactivity, the cellular form of anabena, without considerable change, does not morph;
See Figure 10, contrast reference example 2, in embodiment 4-6, the increment of raw meat algae is relatively many, and the anabena promoter action in embodiment 4 is the most obvious, embodiment 5 and embodiment 6 promoter action relatively little;
See table 2, for cultivating the total lipid content of rear raw meat algae in reference examples 2 and embodiment 4-6, the fat content of relative comparison example 2, the fat content of embodiment 5 and embodiment 6 increases on a small quantity, and the fat content of embodiment 4 increases considerably, and is reference examples about two times.
The different uranium ore material concentration of table 2 is on the impact of Anabaena Flos-aquae fat content
Numbering Reference examples 2 Embodiment 4 Embodiment 5 Embodiment 6
Fat content (%) 8.97 20.63 10.44 9.39
Above embodiment illustrates, blue-green algae substratum, can make blue-green algae oil quantity while normal proliferative increase substantially.

Claims (10)

1. a blue-green algae substratum, is characterized in that, this blue-green algae substratum comprises Bg11 substratum and uranous uranic oxide, and wherein, with the total mass of Bg11 substratum for benchmark, the mass concentration of described uranous uranic oxide is 1ppm-100ppm.
2. blue-green algae substratum according to claim 1, is characterized in that, with the total mass of Bg11 substratum for benchmark, the mass concentration of described uranous uranic oxide is 3ppm, 15ppm and 75ppm.
3. blue-green algae substratum according to claim 1, is characterized in that, the pH value of described Bg11 substratum is 5-9.
4. the blue-green algae substratum according to claim 1 or 3, is characterized in that, the pH value of described Bg11 substratum is 6-8.
5. blue-green algae substratum according to claim 1, is characterized in that, intensity of illumination when this blue-green algae substratum is specifically for cultivating blue-green algae is 10 μm of ol/m 2/ s-50 μm of ol/m 2/ s.
6. blue-green algae substratum according to claim 1 or 5, is characterized in that, intensity of illumination when this blue-green algae substratum is specifically for cultivating blue-green algae is 20 μm of ol/m 2/ s-30 μm of ol/m 2/ s.
7. blue-green algae substratum according to claim 1, is characterized in that, envrionment temperature when this blue-green algae substratum is specifically for cultivating blue-green algae is 10 DEG C-40 DEG C.
8. the blue-green algae substratum according to claim 1 or 7, is characterized in that, envrionment temperature when this blue-green algae substratum is specifically for cultivating blue-green algae is 25 DEG C-27 DEG C.
9. verify a method for the impact that radioactive material confrontation blue-green algae is cultivated, it is characterized in that, the method comprises the following steps:
In Bg11 substratum, add uranous uranic oxide, to the blue-green algae obtained carry out the speed of growth and and the test of Organic Matter Enrichment, wherein, with the total mass of Bg11 substratum for benchmark, the addition of described uranous uranic oxide is 1ppm-100ppm.
10. the method for the impact of checking radioactive material confrontation blue-green algae cultivation according to claim 9, it is characterized in that, with the total mass of Bg11 substratum for benchmark, the addition of described uranous uranic oxide is 3ppm, 15ppm and 75ppm.
CN201511023126.1A 2015-12-30 2015-12-30 Cyanobacteria culture medium Pending CN105483046A (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103882093A (en) * 2012-12-22 2014-06-25 青岛中仁药业有限公司 Method for inhibiting blue algae by using ferulic acid
CN104211146A (en) * 2014-09-15 2014-12-17 深圳职业技术学院 Method for inhibiting blue algae from growth by using plant fallen leaves

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103882093A (en) * 2012-12-22 2014-06-25 青岛中仁药业有限公司 Method for inhibiting blue algae by using ferulic acid
CN104211146A (en) * 2014-09-15 2014-12-17 深圳职业技术学院 Method for inhibiting blue algae from growth by using plant fallen leaves

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
唐东山 等: "铀胁迫对两种蓝藻生长及抗氧化酶活性的影响", 《生态毒理学报》 *
宋东辉 等: "生物柴油原料资源高油脂微藻的开发利用", 《生物工程学报》 *
张荣芳: "铀胁迫对两种蓝藻生长及抗氧化酶活性的影响", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 *
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Application publication date: 20160413