CN114181850B - UV radiation resistant bacillus and application thereof - Google Patents
UV radiation resistant bacillus and application thereof Download PDFInfo
- Publication number
- CN114181850B CN114181850B CN202111368182.4A CN202111368182A CN114181850B CN 114181850 B CN114181850 B CN 114181850B CN 202111368182 A CN202111368182 A CN 202111368182A CN 114181850 B CN114181850 B CN 114181850B
- Authority
- CN
- China
- Prior art keywords
- bacillus
- radiatus
- radiation
- strain
- similarity
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
-
- 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
- C12N1/02—Separating microorganisms from their culture media
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Tropical Medicine & Parasitology (AREA)
- Veterinary Medicine (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Microbiology (AREA)
- Dermatology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a novel strain of Bacillus radiatus (Radioactive sp.), and the Bacillus radiatus (Radioactive sp.) 80 with the collection number of GDMCC 61438 is provided. The bacillus radiatus (Radiobacillus sp.) 80 provided by the invention can be directly used as a gene donor of UV radiation resistant transgenic organisms, and an excellent resistance gene of the bacillus radiatus can be transferred into other organisms, so that the bacillus radiatus can obtain an excellent stress resistance character. And meanwhile, the method can also be used for developing high UV radiation protection articles. Has important significance for researching the UV resistance mechanism of the DNA and promoting the development of new DNA technology in environmental protection, biological repair, human health and the like.
Description
Technical Field
The disclosure relates to the technical field of microorganisms, in particular to a strain of bacillus resistant to UV radiation and the technical field of application thereof.
Background
Suitable environmental conditions are a prerequisite for good plant growth, which is inhibited when the plant is subjected to abiotic stresses (radiation, oxidation, high salt, high osmotic pressure, high temperature, etc.). The UV radiation causes plant dwarfing, leaf thickening, reduction of photosynthetic rate, removal of apical dominance and uneven growth of plant organs, and directly causes the reduction of crop yield and quality.
The strain with the ultraviolet radiation resistance is screened, the related functional genes are identified and then transformed into the plant, the plant with the stress resistance is favorably cultivated, so that the strain is suitable for the external continuously changing climatic environment, and the method has important practical significance.
The Takrama dry desert has the advantages of severe natural climate conditions, large day and night temperature difference, extreme drought, windiness and high ultraviolet irradiation, is a typical extreme environment, has some extreme microbial populations, is not in a reclamation stage, and is a treasure for discovering new microbial resources with special functions. New microbial species isolated from desert samples are continuously reported, such as Jianglella gansuensis, lechevalieria xinjiangensis, planobacterium taklimakanensis, desertibacter roseus, yuhushiella deserti, siccubicoccus deserti, deinococcus malanensis and Radiobacterium deserti, most of which show better stress resistance, especially salt resistance and radiation resistance, and provide new genetic information and special metabolic pathways for the development of molecular biology, genetics, bioinformatics and bioengineering technologies.
Currently, researches on radiation-resistant bacteria and transformation of functional genes thereof are mostly focused on Deinococcus radiodurans.
Disclosure of Invention
The invention aims to separate and culture a new strain from a tacrarma dry desert soil sample, separate a new microorganism with UV radiation resistance, namely a radiating bacillus (Radiobacillus sp.) 80, and can be used for developing high UV radiation protection products. Has important significance for researching the UV resistance mechanism of the DNA and promoting the development of new DNA technology in environmental protection, biological repair, human health and the like.
The purpose of the invention is realized by the following technical scheme:
the technical scheme of the invention is as follows:
the invention provides a bacillus radiata with a preservation number of GDMCC NO:61438, wherein the bacillus radiata is bacillus radiata sp.80 resistant to UV radiation.
The invention also provides application of the bacillus radiatus as a strain with UV radiation resistance function.
The Bacillus radiobacter 80 of the present invention is a gram-positive bacterium of the genus Radiobacterium. By researching the UV resistance mechanism, the understanding of the generation mechanism of the UV resistance of people can be further deepened, and a new transgenic organism is constructed to obtain other excellent characters.
The bacillus radiatus (radio bacillus sp.) 80 provided by the invention is obtained by separating and screening a tacremor sand desert soil sample, the strain number is 80, and the bacillus radiatus (radio bacillus sp.) 80 is determined to be a potential new strain through microbial taxonomic detection and identification. At present, this strain has been deposited in the international collection of microorganisms under the Budapest treaty before the filing date: guangdong province culture collection of microorganisms (GDMCC). Address: address: the preservation date of the No. 5 storied building of No. 59 Dazhong 100 Jie of Guangzhou city is 2021 year, 1 month and 15 days, and the preservation number is GDMCC NO:61438.
The bacillus radiatus sp potential new strain 80 provided by the invention is gram-positive, the cell is long rod-shaped, the size is 0.25-0.28 μm wide, 1.4-4.9 μm long, the tail end generates expanded sporangium and endogenetic spore, the size of the spore is 0.6-0.7 x 1.0-1.4 μm, and after the bacillus radiatus sp grows on a culture medium and is cultured, the colony is milky and convex, round, opaque, the surface is moist and smooth, and the edge is tidy.
The 16S rDNA sequence of the potential new strain 80 of the bacillus radiatus (radio bacillus sp.) is amplified by taking the genome total DNA of the potential new strain as a template, sequencing is carried out, the 16S rDNA sequence (shown in a sequence table) with the length of 1436bp is obtained, homology comparison analysis is carried out on the sequence and related strains recorded in a GenBank database, and the similarity of the strain and the other strain of the same genus is 98.68 percent. And the same genus of Radiobacillus deserti TKL69 T Similarity 98.92%; the similarity with the genus Aquibacillus is 96.83% -95.88%, and the similarity with Aquibacillus halophilus B6B T Has a similarity of 96.83%; the similarity with the genus Salinibacillus is 96.61-95.21%, and the Salinibacillus xinjiangensis J4 T The similarity of (a) is 96.61%; the similarity with Ornithia sp is 96.61-95.05%, and the similarity with Ornithia salinisisoli LCB256T is 96.61%; the similarity with Sedimiibacillus is 96.23-95.77%, and the obtained sequence is commonAnd carrying out comparison analysis on the NCBI website, constructing a phylogenetic tree, and primarily determining the phylogenetic tree as a potential new species, wherein the name of a tentative strain is bacillus radiatus (radio bacillus sp.) potential new strain 80.
The bacillus radiatus sp potential new strain 80 is cultured by a liquid culture medium prepared by adding 1.5% of agar to a liquid culture medium or a solid culture medium prepared by adding 1.5% of peptone, 1.0g of yeast powder, 0.1g of ferric citrate, 19.45g of sodium chloride, 5.98g of magnesium chloride, 3.24g of sodium sulfate, 1.8g of calcium chloride, 0.55g of potassium chloride, 0.16g of sodium carbonate, 0.08g of potassium bromide, 0.034g of strontium chloride, 0.022g of boric acid, 0.004g of sodium silicate, 0.0024g of sodium fluoride, 0.0016g of sodium nitrate, 0.008g of disodium hydrogen phosphate, 1000ml of distilled water and 7.6 +/-0.2 of pH value, 120 ℃ and sterilizing for 15min.
The bacillus radiatus sp potential new strain 80 can grow in the temperature range of 15-45 ℃.
The bacillus radiatus sp (radio bacillus sp.) potential new strain 80 can grow in the range of 0-14% of NaCl concentration.
The bacillus radiatus sp (radio bacillus sp.) potential new strain 80 of the invention can grow with the pH value of 6.0-9.0.
The bacillus radiatus (Radiobacillus sp.) 80 can tolerate 84000 mu W.s/cm 2 A dose of radiation.
Further, the invention also provides application of the bacillus radiatus (radio bacillus sp.) potential new strain 80 in the development of UV radiation protection products.
Further, the invention also provides application of the bacillus radiatus (radio bacillus sp.) potential new strain 80 in preparation of anti-UV transgenic organisms.
Further, the invention also provides application of the bacillus radiatus (radio bacillus sp.) potential new strain 80 in preparation of anti-UV transgenic organisms, and the application is a gene donor.
The invention has the following beneficial technical effects:
(1) The bacillus radiatus (radio bacillus sp.) potential new strain 80 has good UV resistance, the survival rate reaches 21.95 percent after 10min of ultraviolet irradiation, and the bacillus radiatus potential new strain is compared with a positive control Deinococcus radiodurans DSM20539 T The survival rate was comparable (22.83%), and the negative control bacterium Escherichia coli BL21 was only 0.76% under the same conditions. After 20min of uv irradiation, 3.3% of the strain 80 was still alive, 4.92% of d.radiodurans DSM20539T, and all the escherichia coli died. The UV-resistant bacillus 80 of the invention is shown to have better UV resistance.
(2) The potential new strain 80 of the bacillus radiatus (radio bacillus sp.) can be directly used as a gene donor of an anti-UV transgenic organism, and an excellent resistance gene of the bacillus radiatus can be transferred into other organisms, so that the bacillus radiatus can obtain an excellent stress resistance character; meanwhile, the strain can also be used as a new genetic engineering strain, and other excellent genes are received to obtain more excellent characters; in addition, by improving the characteristics by the traditional mutation breeding method, more strains with excellent characteristics can be obtained. The bacillus radians 80 of the present invention can be used for the development of high UV radiation protective articles. Has important significance for researching the UV resistance mechanism of the DNA and promoting the development of new DNA technology in environmental protection, biological repair, human health and the like.
Drawings
FIG. 1 is a diagram showing the evolutionary analysis (based on 16S rDNA sequence analysis) between UV-resistant Bacillus 80 and other members of the genus and close to the genus of Bacillus;
FIG. 2 shows a comparison of UV-radiation resistant Bacillus 80 and the control strain Deinococcus radiodurans DSM20539 T Comparison graph of UV resistance of Escherichia coli DL 21;
FIG. 3 is a diagram showing the colony morphology of the UV-resistant Bacillus sp.80;
FIG. 4 shows a transmission electron micrograph of a UV-resistant Bacillus sp.80.
Detailed Description
In order to better explain the invention, the following further illustrate the main content of the invention in connection with specific examples, but the content of the invention is not limited to the following examples. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are commercially available products.
The reagents adopted in the invention are as follows: peptone, yeast powder, ferric citrate, sodium chloride, magnesium chloride, sodium sulfate, calcium chloride, potassium chloride, sodium carbonate, potassium bromide, strontium chloride, boric acid, sodium silicate, sodium fluoride, sodium nitrate, disodium hydrogen phosphate and distilled water. Casein hydrolysate, yeast extract, tryptone, glucose, soluble starch, dipotassium hydrogen phosphate, anhydrous magnesium sulfate, sodium pyruvate, sodium chloride and agar which are common reagents in the market, and 2 XPCR Green Mix is purchased from Changsheng biotechnology Limited liability company in Beijing Ding Guo.
The instrument adopted in the invention is as follows: a high-temperature sterilization pot, an electronic balance, a sterile operating platform, a Biolog strain identifier GNIII test board, a PCR instrument, a micropipettor, a gun head, a centrifuge tube, an electrophoresis instrument, a gel imager, a pH meter, a spectrophotometer, an electron microscope, a culture dish, a glass rod, an alcohol lamp, an inoculating loop, a centrifuge and a microwave oven.
The reagents and materials can be purchased through public channels, and the equipment and instruments adopted in the process are common equipment in the field.
All materials, reagents and equipment selected for use in the present invention are well known in the art, but do not limit the practice of the invention, and other reagents and equipment well known in the art may be suitable for use in the practice of the following embodiments of the invention.
The following examples further illustrate the present invention but are not to be construed as limiting the invention. Modifications or substitutions to methods, steps or conditions of the present invention may be made without departing from the spirit and scope of the invention.
The first embodiment is as follows: isolation of Bacillus radiobacter (Radiobacillus sp.) 80
And collecting soil samples in the dry desert of the Tara Karma in Xinjiang. Taking about 5.0g of soil sample, adding 45mL of sterile water, carrying out enrichment culture at 30 ℃ and 200rpm for 30min, taking 1.0mL of enrichment culture, sequentially diluting the enrichment culture in a gradient manner according to 10 times, respectively taking 100 mu L of the enrichment culture, coating the 100 mu L of the enrichment culture on an R2A agar medium plate containing 1 percent of NaCl, and inversely placing the enrichment culture on an incubator at 30 ℃ for culturing for 15 days to obtain the UV radiation resistant bacillus.
R2A agar medium: 0.5g/L of casein hydrolysate; yeast extract 0.5g/L; tryptone 0.5g/L; glucose 0.5g/L; 0.5g/L of soluble starch; dipotassium hydrogen phosphate 0.3g/L; anhydrous magnesium sulfate 0.024g/L; 0.3g/L of sodium pyruvate; 10g/L of sodium chloride; 15g/L of agar; pH =7.2 ± 0.2. Sterilizing at 121 deg.C for 15min. The culture temperature was 30 ℃.
Example two: identification of Bacillus radiatus (Radiobacillus sp.) 80
The culture conditions of the UV radiation resistant bacillus (Radiobacillus sp.) 80 are as follows:
adopting 2216E agar culture medium, the concrete formula is: 5.0g of peptone, 1.0g of yeast powder, 0.1g of ferric citrate, 19.45g of sodium chloride, 5.98g of magnesium chloride, 3.24g of sodium sulfate, 1.8g of calcium chloride, 0.55g of potassium chloride, 0.16g of sodium carbonate, 0.08g of potassium bromide, 0.034g of strontium chloride, 0.022g of boric acid, 0.004g of sodium silicate, 0.0024g of sodium fluoride, 0.0016g of sodium nitrate, 0.008g of disodium hydrogen phosphate and 1000ml of distilled water. The pH value is 7.6 +/-0.2; the culture temperature is as follows: 37 ℃ is carried out.
The UV radiation-resistant Bacillus sp (Radiobacillus sp.) 80 of the present invention has the following properties:
1. morphological characteristics
As can be seen by a transmission electron microscope, the cells are long rod-shaped, the size is 0.25-0.28 μm wide,
1.4-4.9 μm long. The end generates expanded sporangium and endogenic spore with size of 0.6-0.7 × 1.0-1.4 μm. As shown in fig. 4.
2. Characteristics on various media:
on 2216E agar medium, the medium is milky white, convex, round, opaque, moist, smooth in surface, and neat in edge, as shown in FIG. 3.
3. Physiological and biochemical characteristics
The strain is aerobically grown, gram-positive, positive in catalase, oxidase, nitrate reductase and beta-galactosidase, hydrolyzes Tween 20, liquefies gelatin,does not generate H 2 S, using D-mannose, cellobiose, turanose, D-galactose and L-histidine as a single carbon source. The cell has main peptidoglycan including DPG, APL, PL and L, and main fatty acid including anteiso-C 15:0 、iso-C 15:0 、anteiso-C 17:0 The predominant respiratory quinone is MK-7.
4. Carbon source utilization
As shown in Table 1, glycerol, L-arabinose, ribose, D-xylose, galactose, glucose, fructose, mannose, sorbose, rhamnose, mannitol, α -methyl-D-glucoside, N-acetyl-glucosamine, amygdalin, arbutin, esculetin, saligenin, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, inulin, melezitose, raffinose, starch, glycogen, gentiobiose, D-turanose, D-tagatose, and gluconate can be used as the carbon source, but erythritol, D-arabinose, L-xylose, adonitol, β -methyl-D-xyloside, dulcitol, inositol, sorbitol, α -methyl-D-mannoside, xylitol, D-lyxose, D-fucose, L-fucose, D-arabitol, L-arabitol, and 2-keto-gluconate can be used as the carbon source.
5. Other properties
The optimum growth temperature is 35 ℃. The optimum salinity is 2% -4%, and the optimum pH value is 6.0-8.5. Table 1: UV-radiation-resistant bacillus 80 and Radiobacillus desurtitkl 69 T Comparison of microbiological Properties of
Example three: 16S rDNA sequence identification
1. Method for extracting genome
The UV-radiation-resistant bacillus (Radiobacterium sp.) 80 of the present invention is extracted by using a bacterial genomic DNA extraction kit (DP 302) from TIANGEN corporation, as described in the specification.
PCR amplification method
The general primer sequence used for the PCR amplification of the 16S rDNA of the UV-radiation-resistant bacillus (radio bacillus sp.) 80 is synthesized by biological engineering (Shanghai) GmbH. 2 XPCR Green Mix for amplification was purchased from Changsheng Biotechnology, inc., beijing ancient China.
Forward primer 27F:5'-AGAGTTTGATCATGGCTCAG-3' of the formula,
reverse primer 1492R:5'-TACGGTTACCTTGTTACGACTT-3'.
The PCR amplification system is as follows: 50 μ L,2 XPCR Green Mix12.5 μ L, 1 μ L each of the upstream and downstream primers, 1 μ L of the DNA template, 9.5uL of ddH 2O.
PCR reaction procedure: 95 ℃ for 5min;30 cycles (95 ℃,30s, 51 ℃,30s, 72 ℃ for 1.5 min; 72 ℃ for 5min.
The 16S rDNA sequence obtained by amplification was sequenced by the company, weituo engineering bioengineering (Shanghai). The sequencing result comprises: the 16S rDNA sequence is shown in SEQ ID NO 1. The similarity of the 16S rDNA of the UV-radiation-resistant bacillus (radio bacillus sp 80) and the other bacterium of the same genus is 98.68 percent. With the same genus of Radiobacillus deserti TKL69 T Similarity 98.92%; the similarity with the genus Aquibacillus is 96.83% -95.88%, and the similarity with Aquibacillus halophilus B6B T Has a similarity of 96.83%; the similarity with the genus Salinibacillus is 96.61-95.21%, and the Salinibacillus xinjiangensis J4 T The similarity of (a) is 96.61%; the similarity with Ornithiacilli genus is 96.61% -95.05%, and the similarity with Ornithiacilli salinisinoli LCB256T is 96.61%; the similarity with Sedimiibacillus is 96.23-95.77%; the similarity with Salirhabdus is 96.14-95.77%.
Through 16S rDNA sequence similarity, 80 can be determined as a strain of the genus Bacillus radians, and further through whole genome sequencing and average nucleic acid identity (ANI) and dDDH analysis of the sequence, the ANI value of 80 and another standard mean TKL69T of the genus Bacillus radians are 78.39% (< 95%), the dDDH value is 22% (< 70%), and meanwhile, the physiological and biochemical property experimental results of the two are obviously different, which indicates that 80 is a new species of the genus Bacillus radians.
The inventor separates and cultivates a new strain from a Takrama dry desert soil sample, the strain is identified and confirmed to belong to the genus Bacillus radiobacter and is named as UV radiation resistant Bacillus (Radiobacillus sp.) 80, the strain is preserved in Guangdong province microbial strain preservation center of preservation unit appointed by the State intellectual property office, the preservation date is 1 month and 15 days at 2021 year, and the preservation registration number is GDMCC NO:61438.
Example four: UV radiation resistant Bacillus 80 UV radiation resistant test
1. Test method
(1) Selecting the UV-radiation-resistant bacillus glycerol storage tube bacterial liquid, carrying out plate streaking activation on a 2216E culture medium, and carrying out inverted culture at 30 ℃;
(2) Picking colonies on the plate, inoculating the colonies into 2216E liquid culture medium, performing shaking culture at 30 ℃ and 200rpm for 3d;
(3) Determination of OD 600 According to the object OD 600 = 0.1-0.2, adding into fresh 2216E liquid culture medium, culturing at 30 deg.C and 200rpm to OD 600 0.6 to 0.8;
(4) Adjusting the OD of the bacterial suspension with sterile water 600 Is 0.5. Note that OD was adjusted 600 When the culture medium is used, the corresponding non-inoculated culture medium is used as a blank control.
(5) 1ml of OD was taken 600 0.5 bacteria solution, placed in a 35mm petri dish (petri dish was opened under an ultraviolet lamp in advance, and the cover was kept open during the subsequent irradiation process), and irradiated under an ultraviolet lamp for 10min and 20min.
(6) The irradiated bacterial liquid quickly dilutes each sample to 10 -5 、10 -6 Then, 100. Mu.L of the solution was applied to a 2216E plate. Incubated at 37 ℃ for 5 days and counted.
Simultaneously respectively using Deinococcus radiodurans DSM20539 T And Escherichia coli BL21 strain as positive control and negative control, respectively culturing and irradiating in TGY and LB culture medium with 253.7nm and 20W ultraviolet lamp at 70 μ W/cm intensity 2 The dose of irradiation for 10min and 20min was 42000. Mu.W.s/cm, respectively 2 And 84000. Mu. Ws/cm 2 . The test and control bacteria were each subjected to 3 replicates.
2. Results
The experimental result shows that after 10min ultraviolet irradiation, the survival rate reaches 21.95 percent, and the survival rate is compared with positive control Deinococcus radiodurans DSM20539 T The survival rate was comparable (22.83%), and the negative control bacterium Escherichia coli BL21 was only 0.76% under the same conditions. After 20min uv irradiation, 3.3% of the strain 80 was still alive, 4.92% of d.radiodurans DSM20539T, and all the escherichia coli died. The UV-resistant bacillus 80 of the invention is shown to have better UV resistance. The UV-resistant bacillus 80 of the present invention can be used for the development of UV radiation protective products such as protective coatings and related cosmetics. The main sunscreen components of the existing sunscreen product are zinc oxide and titanium oxide, which are chemical components, and the UV radiation resistant bacillus 80 can be used for developing a sunscreen product containing biological components.
Example five: extraction of UV radiation resistant Bacillus 80 strain genome DNA
The invention also provides a genome extraction and 16S rDNA amplification method of the UV-radiation-resistant bacillus (radio bacillus sp.) 80, which is specifically carried out according to the following operations:
1. method for extracting genome
The UV-radiation-resistant bacillus 80 is extracted by using a bacterial genome DNA extraction kit (DP 302) of TIANGEN company, and the details are shown in the specification.
PCR amplification method
The general primer sequence for the PCR amplification of the 16S rDNA of the UV-radiation-resistant bacillus 80 is synthesized by the company of Biotechnology engineering (Shanghai). 2 XPCR Green Mix for amplification was purchased from Changsheng Biotechnology, inc., beijing ancient China.
Forward primer 27F:5'-AGAGTTTGATCATGGCTCAG-3' of the formula,
reverse primer 1492R:5'-TACGGTTACCTTGTTACGACTT-3'.
The PCR amplification system is as follows: 50 μ L,2 XPCR Green Mix12.5 μ L, 1 μ L each of the upstream and downstream primers, 1 μ L of the DNA template, 9.5uL of ddH 2O.
PCR reaction procedure: 95 ℃ for 5min;30 cycles (95 ℃,30s, 51 ℃,30s, 72 ℃ for 1.5 min; 72 ℃ for 5min.
Namely, the 16S rDNA sequence of the bacillus radiatus (Radiobacterium sp) 80 obtained by amplification is obtained by amplification, the sequencing result of the sequence with the size of 1436bp and 1696S rDNA is shown as SEQ ID NO: 1.
The preferred embodiments of the present disclosure are described in detail with reference to the accompanying drawings, however, the present disclosure is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present disclosure within the technical idea of the present disclosure, and these simple modifications all belong to the protection scope of the present disclosure.
It should be noted that, in the foregoing embodiments, various features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various combinations that are possible in the present disclosure are not described again.
In addition, any combination of various embodiments of the present disclosure may be made, and the same should be considered as the disclosure of the present disclosure, as long as it does not depart from the spirit of the present disclosure.
Sequence listing
<110> institute for the application of microorganisms of Sinkiang academy of agricultural sciences (Xinjiang-Yameinian bioengineering research and development center, china)
<120> UV radiation resistant bacillus and application thereof
<141> 2021-11-18
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1436
<212> DNA
<213> Bacillus radiatus 80 (Radiobacillus sp.80)
<400> 1
tatacatgca gtcgagcgcg ggaagccatc tgaccccttc ggggtgacga tggtggaacg 60
agcggcggac gggtgagtaa cacgtgggta acctgcctat aagactggga taactccggg 120
aaaccggggc taataccgga taatactttt tcctgcatgg gagaaagttg aaaggcggct 180
tcggctgtcg cttatagatg gacccgcggc gcattagcta gttggtaggg taatggccta 240
ccaaggcaac gatgcgtagc cgacctgaga gggtgatcgg ccacactggg actgagacac 300
ggcccagact cctacgggag gcagcagtag ggaatcttcc gcaatggacg aaagtctgac 360
ggagcaacgc cgcgtgaacg atgaaggttt tcggatcgta aagttctgtt gttagggaag 420
aacaagtaca agagtaactg cttgtacctt gacggtacct gacgagaaag ccccggctaa 480
ctacgtgcca gcagccgcgg taatacgtag ggggcaagcg ttgtccggaa ttattgggcg 540
taaagcgcgc gcaggcggtt tcttaagtct gatgtgaaag cccacggctt aaccgtggag 600
ggtcattgga aactggggaa cttgagtgca gaagaggaga gtggaattcc acgtgtagcg 660
gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag gcgactctct ggtctgtaac 720
tgacgctgag gcgcgaaagc gtggggagcg aacaggatta gataccctgg tagtccacgc 780
cgtaaacgat gagtgctagg tgttaggggg tttccgcccc ttagtgctga agttaacgca 840
ttaagcactc cgcctgggga gtacggccgc aaggctgaaa ctcaaaagaa ttgacggggg 900
cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt 960
cttgacatcc tctgarcact ctagagatag agctttccct tcggggacag agtgacaggt 1020
ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc 1080
aacccttgat cttagttgcc agcattttag ttgggcactc taaggtgact gccggtgaca 1140
aaccggagga aggtggggat gacgtcaaat catcatgccc cttatgacct gggctacaca 1200
cgtgctacaa tggatggtac aaagggcagc gaagccgcga ggtgaagcaa atcccataaa 1260
accattctca gttcggattg taggctgcaa ctcgcctaca tgaagccgga atcgctagta 1320
atcgcggatc agcatgccgc ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac 1380
accacgagag ttggcaacac ccgaagtcgg tgaggtaacc ttttaggagc cagccg 1436
Claims (2)
1. A strain of Bacillus radiatus (Radiobacillus sp.) 80, characterized in that said bacillus radiatus (b), (b)Radiobacillus sp.) 80 under the accession number GDMCC NO 61438.
2. The Bacillus radiobacter of claim 1 (A)Radiobacillus sp.) Use of 80 strains in the development of articles for protection against ultraviolet radiation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111368182.4A CN114181850B (en) | 2021-11-18 | 2021-11-18 | UV radiation resistant bacillus and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111368182.4A CN114181850B (en) | 2021-11-18 | 2021-11-18 | UV radiation resistant bacillus and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114181850A CN114181850A (en) | 2022-03-15 |
| CN114181850B true CN114181850B (en) | 2023-01-31 |
Family
ID=80540320
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111368182.4A Active CN114181850B (en) | 2021-11-18 | 2021-11-18 | UV radiation resistant bacillus and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114181850B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117448242B (en) * | 2023-12-20 | 2024-04-30 | 航天神舟生物科技集团有限公司 | Bacillus licheniformis and application thereof in field of planetary protection |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63317078A (en) * | 1987-06-18 | 1988-12-26 | Mitsubishi Kasei Corp | Novel microorganism |
| JP2008228572A (en) * | 2007-03-16 | 2008-10-02 | Akita Prefectural Univ | Rhizobium |
| CN101481672A (en) * | 2009-01-22 | 2009-07-15 | 广东省微生物研究所 | Radiation resistant microbacterium and use thereof |
| CN103911318A (en) * | 2014-03-07 | 2014-07-09 | 中国农业科学院生物技术研究所 | Radiation resistant Rhodobacter shamoensis W402, and applications thereof |
| CN104974957A (en) * | 2015-06-26 | 2015-10-14 | 滨州学院 | Bacillus (bacillus sp.) ZY bacterial strain and application thereof |
| CN107325980A (en) * | 2017-06-12 | 2017-11-07 | 新疆农业科学院微生物应用研究所（中国新疆-亚美尼亚生物工程研究开发中心） | A kind of radiation hardness series bacillus KH9 and its application in biological antitranspirant |
| CN109182215A (en) * | 2018-10-17 | 2019-01-11 | 湖南省林业科学院 | One bacillus licheniformis bacterial strain and its application in inhibition bacterial pathogen |
| CN112662580A (en) * | 2019-11-27 | 2021-04-16 | 中国农业科学院生物技术研究所 | Paenibacillus aerophilus and application thereof |
-
2021
- 2021-11-18 CN CN202111368182.4A patent/CN114181850B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63317078A (en) * | 1987-06-18 | 1988-12-26 | Mitsubishi Kasei Corp | Novel microorganism |
| JP2008228572A (en) * | 2007-03-16 | 2008-10-02 | Akita Prefectural Univ | Rhizobium |
| CN101481672A (en) * | 2009-01-22 | 2009-07-15 | 广东省微生物研究所 | Radiation resistant microbacterium and use thereof |
| CN103911318A (en) * | 2014-03-07 | 2014-07-09 | 中国农业科学院生物技术研究所 | Radiation resistant Rhodobacter shamoensis W402, and applications thereof |
| CN104974957A (en) * | 2015-06-26 | 2015-10-14 | 滨州学院 | Bacillus (bacillus sp.) ZY bacterial strain and application thereof |
| CN107325980A (en) * | 2017-06-12 | 2017-11-07 | 新疆农业科学院微生物应用研究所（中国新疆-亚美尼亚生物工程研究开发中心） | A kind of radiation hardness series bacillus KH9 and its application in biological antitranspirant |
| CN109182215A (en) * | 2018-10-17 | 2019-01-11 | 湖南省林业科学院 | One bacillus licheniformis bacterial strain and its application in inhibition bacterial pathogen |
| CN112662580A (en) * | 2019-11-27 | 2021-04-16 | 中国农业科学院生物技术研究所 | Paenibacillus aerophilus and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114181850A (en) | 2022-03-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Castillo et al. | Halovivax asiaticus gen. nov., sp. nov., a novel extremely halophilic archaeon isolated from Inner Mongolia, China | |
| Reddy et al. | Description of Hymenobacter arizonensis sp. nov. from the southwestern arid lands of the United States of America | |
| Lee et al. | Spirosoma radiotolerans sp. nov., a gamma-radiation-resistant bacterium isolated from gamma ray-irradiated soil | |
| Gorlas et al. | Thermococcus prieurii sp. nov., a hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent | |
| CN114181850B (en) | UV radiation resistant bacillus and application thereof | |
| Cha et al. | Deinococcus soli sp. nov., a gamma-radiation-resistant bacterium isolated from rice field soil | |
| CN110272850B (en) | New strain with algae dissolving capacity and application thereof to phaeocystis globosa | |
| Li et al. | Roseovarius marisflavi sp. nov., isolated from an amphioxus breeding zone in the coastal region of the Yellow Sea, China | |
| Wang et al. | Pseudomonas rhizoryzae sp. nov., isolated from rice | |
| CN113564081B (en) | Devorax SCS-3 for producing vomitoxin degrading enzyme and application thereof | |
| Doronina et al. | Ancylobacter moscoviensis sp. nov., novel facultatively methylotrophic bacteria from activated sludge and the reclassification of Starkeya novella (Starkey 1934) Kelly et al. 2000 as Ancylobacter novellus comb. nov., Starkeya koreensis Im et al. 2006 as Ancylobacter koreensis comb. nov., Angulomicrobium tetraedrale Vasil'eva et al. 1986 as Ancylobacter tetraedralis comb. nov., Angulomicrobium amanitiforme Fritz et al. 2004 as Ancylobacter amanitiformis comb. nov., and Methylorhabdus multivorans Doronina et al. 1996 as Ancylobacter multivorans comb. nov., and emended description of the genus Ancylobacter | |
| Márquez et al. | Aquisalibacillus elongatus gen. nov., sp. nov., a moderately halophilic bacterium of the family Bacillaceae isolated from a saline lake | |
| Li et al. | Spirosoma pomorum sp. nov., isolated from apple orchard soil | |
| Wang et al. | Flammeovirga agarivorans sp. nov., an agar-digesting marine bacterium isolated from surface seawater | |
| CN112662580B (en) | Paenibacillus aerophilus and application thereof | |
| Su et al. | Rhizobium flavescens sp. nov., isolated from a chlorothalonil-contaminated soil | |
| Lee et al. | Phaeobacter marinintestinus sp. nov., isolated from the intestine of a sea cucumber (Apostichopus japonicus) | |
| Ten et al. | Hymenobacter pomorum sp. nov., isolated from apple orchard soil | |
| Liu et al. | Phytohalomonas tamaricis gen. nov., sp. nov., an endophytic bacterium isolated from Tamarix ramosissima roots growing in Kumtag desert | |
| Liu et al. | Streptomyces polyasparticus sp. nov. isolated from cotton field soil by a medium applied with polyaspartic acid | |
| Lee et al. | Deinococcus knuensis sp. nov., a bacterium isolated from river water | |
| CN108148788B (en) | Antioxidant staphylococcus stewardii W17 and application thereof | |
| Wang et al. | Lysobacter lactosilyticus sp. nov., a Novel β-Galactosidase Producing Bacterial Strain Isolated from Farmland Soil Applied with Amino Acid Fertilizer | |
| Ten et al. | Deinococcus terrigena sp. nov., a novel member of the family Deinococcaceae | |
| Lee et al. | Paenibacillus swuensis sp. nov., a bacterium isolated from soil |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |