CN112458024B - Brevundimonas deep sea and application thereof in induction of adhesion of mytilus coruscus - Google Patents
Brevundimonas deep sea and application thereof in induction of adhesion of mytilus coruscus Download PDFInfo
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- CN112458024B CN112458024B CN202011469883.2A CN202011469883A CN112458024B CN 112458024 B CN112458024 B CN 112458024B CN 202011469883 A CN202011469883 A CN 202011469883A CN 112458024 B CN112458024 B CN 112458024B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/50—Culture of aquatic animals of shellfish
- A01K61/54—Culture of aquatic animals of shellfish of bivalves, e.g. oysters or mussels
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Abstract
The invention discloses deep sea shortwave monad and application thereof in inducing adhesion of thick shell mussels, belonging to the technical field of marine microorganisms. The Brevundimonas abyssal is named Brevundimonas abyssal sp.c7200a and is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M2019802. The 16S rRNA gene of the bacterium is shown as SEQ ID NO. 1, and is derived from sea water of the North Pacific Lepidium meyenii at an altitude of-7200 meters. The invention obtains the deep sea shortwave monad for the first time, obtains the deep sea shortwave monad Brevundimonas sp.c7200a through separation, purification and identification, verifies that the biofilm formed by the bacteria has the activity of inducing the adhesion of the thick-shell mussel, and provides a new technical approach for solving the problem of seedling and utilizing the bacteria to form the biofilm to induce the adhesion of the thick-shell mussel.
Description
Technical Field
The invention relates to the technical field of marine microorganisms, in particular to deep-sea shortwave monads and application thereof in inducing adhesion of thick-shell mussels.
Background
Mytilus coruscus (Mytilus coruscus) belongs to the phylum Mollusca (Mollusca), class Bivalvia (Bivalvia), order Mytilodia (Mytilodia), family Mytilidae (Mytilidae), is an important marine shellfish culture variety in China, and is distributed on the coasts of yellow sea, bohai sea and east sea. Mytilus coruscus has two life forms, including planktonic life and adhesive life, and mytilus coruscus larvae adhere to and become young larvae at the later stages of planktonic life, and eventually grow into adult shellfish. In the process of growing the young shellfish into adult shellfish, once the environment changes, the young shellfish of the thick-shell mussel can automatically cut off the podites and re-select a proper attachment base for reattachment.
The mytilus coruscus has wide breeding prospect and market development potential as a seawater shellfish breeding variety with high nutritive value, and the fries are mainly obtained by natural fries and semi-artificial fries collection. However, with the excessive development of fishery resources, natural seedlings cannot meet the development of the mytilus coruscus culture industry, and the development of bioengineering technologies such as solving the seedling problem and utilizing bacteria to form a biofilm to induce the adhesion of mytilus coruscus is urgent.
Disclosure of Invention
The first object of the present invention is to provide a strain of Brevundimonas sp.
The second object of the invention is to provide the application of the deep sea shortwave monad in inducing the adhesion of thick shell mussel.
In order to achieve the first object, the present invention adopts the following technical scheme: the deep-sea Brevundimonas sp.c7200a is named as deep-sea Brevundimonas sp.c7200a, and is preserved in China Center for Type Culture Collection (CCTCC) NO: M2019802, the preservation date is 10 months 11 in 2019, and the preservation address is university of Wuhan in China.
According to an embodiment of the invention, the 16S rRNA gene of Brevundimonas sp.c7200a is shown in SEQ ID NO. 1.
According to an embodiment of the invention, the Brevundimonas sp.c7200a is from sea water (148 ° 53.2473'e,53.9694' n) from the sea ditch of the Pacific ocean, altitude-7200 meters.
According to an embodiment of the invention, the biofilm formed by Brevundimonas deep sea sp.c7200a has the activity of inducing adhesion of mytilus coruscus.
In order to achieve the second object, the present invention adopts the following technical scheme: the application of the deep sea shortwave monad in inducing the adhesion of the thick shell mussel is provided.
According to an embodiment of the invention, the mytilus coruscus is a juvenile mollusk.
By adopting the technical scheme, the invention has the following beneficial effects: the invention obtains the deep sea shortwave monad for the first time, obtains the deep sea shortwave monad Brevundimonas sp.c7200a through separation, purification and identification, verifies that the biofilm formed by the bacteria has the activity of inducing the adhesion of the thick-shell mussel, and provides a new technical approach for solving the problem of seedling and utilizing the bacteria to form the biofilm to induce the adhesion of the thick-shell mussel.
Drawings
FIG. 1 is the induction of adhesion of biofilms formed at different initial bacterial densities to juvenile mytilus coruscus.
Detailed Description
The present invention is further illustrated by the following examples and comparative examples, which are only for illustration of the present invention, and the present invention is not limited to the following examples. All modifications and equivalent substitutions to the technical proposal of the invention are included in the protection scope of the invention without departing from the scope of the technical proposal of the invention.
In the following examples, brevundimonas sp was designated Brevundimonas sp.c7200a, and was deposited at China center for type culture Collection with the accession number CCTCC NO: M2019802, the date of deposit was 2019, 10 months and 11 days, and the deposit address was the university of Wuhan, china.
The 16S rRNA gene of Brevundimonas sp.c7200a in deep sea is shown as SEQ ID NO:1, pure colonies are obtained by separation and purification of Zobell 2216E solid medium, and the pure colonies are uploaded to NCBI database after comparison with nucleic acid data in GENEBANK (http:// www.ncbi.nlm.nih.gov /) through 16S rRNA gene sequencing, and the sequence number is FR754553.1, and the method is as follows:
acacatgcagtcgaacgaactcttcggagttagtggcggacgggtgagtaacacgtgggaacgtgcctttaggttcggaataactcaggga aacttgtgctaataccgaatgtgcccttcgggggaaagatttatcgcctttagagcggcccgcgtctgattagctagttggtgaggtaaaggc tcaccaaggcgacgatcagtagctggtctgagaggatgatcagccacattgggactgagacacggcccaaactcctacgggaggcagca gtggggaatcttgcgcaatgggcgaaagcctgacgcagccatgccgcgtgaatgatgaaggtcttaggattgtaaaattctttcagtaggga cgataatgacggtacctacagaagaagccccggctaacttcgtgccagcagccgcggtaatacgaagggggctagcgttgctcggaatta ctgggcgtaaagggagcgtaggcggacatttaagtcaggggtgaaatcccggggctcaacctcggaattgcctttgatactgggtgtcttg agtgtgagagaggtatgtggaactccgagtgtagaggtgaaattcgtagatattcggaagaacaccagtggcgaaggcgacatactggct cattactgacgctgaggctcgaaagcgtggggagcaaacaggattagataccctggtagtccacgccgtaaacgatgattgctagttgtcg ggatgcatgcatttcggtgacgcagctaacgcattaagcaatccgcctggggagtacggtcgcaagattaaaactcaaaggaattgacggg ggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgcagaaccttaccaccttttgacatgcctggaccgccagagagatc tggctttcccttcggggactaggacacaggtgctgcatggctgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgca accctcgccattagttgccatcatttagttgggaactctaatgggactgccggtgctaagccggaggaaggtggggatgacgtcaagtcctc atggcccttacagggtgggctacacacgtgctacaatggcgactacagagggttaatccttaaaagtcgtctcagttcggattgtcctctgca actcgagggcatgaagttggaatcgctagtaatcgcggatcagcatgccgcggtgaatacgttcccgggccttgtacacaccgcccgtcac accatgggagttggttctacccgaaggcgctgcgctgaccgcaagggggcaggcgaccacg。
in one embodiment, brevundimonas sp.c7200a may be derived from sea water from the sea ditch of Pacific ocean, at an altitude of-7200 meters (148℃53.2473'E,53.9694' N).
The following specific examples demonstrate that the biofilm formed by Brevundimonas deep sea sp.c7200a has activity of inducing adhesion of mytilus coruscus, and can be used for inducing adhesion of mytilus coruscus, especially young shellfish.
Example 1
The experimental method for inducing adhesion of Brevundimonas sp.c7200a to juvenile mollusk of mytilus coruscus comprises the following specific steps: the pure Brevundimonas deep sea sp.c7200a strain is usedExpanding culture in seawater 2216E liquid culture medium for 48 hr, centrifuging to obtain bacterial precipitate, washing bacteria with sterilized seawater, preparing into suspension, repeating for 3 times to obtain bacterial liquid with constant volume of 50mL, staining with acridine orange, randomly selecting 10 points under 100 times lens fluorescent microscope, counting, and determining total bacterial liquid density of 2.5X10 9 cells/mL. Placing a sterilized glass slide in each culture dish, and adding corresponding sterilized seawater and bacterial liquid to make initial bacterial density 1.0X10 respectively 6 cells/mL、1.0×10 7 cells/mL、1.0×10 8 cells/mL and 5X 10 8 culturing cells/mL at 18deg.C under dark condition for 48 hr to form desired biofilm, adding slide glass with biofilm and 10 young shellfish of Mytilus coruscus (shell length 0.80+ -0.05 mm, shell height 0.55+ -0.05 mm) into culture dish containing sterile filtered seawater, and recording adhesion rate of young shellfish at 24 hr, which is the percentage of number of young shellfish adhered on slide glass to total number of young shellfish in culture dish.
As shown in FIG. 1, the biofilm formed under the conditions of different initial bacterial densities has significantly different induced activities on juvenile mollusks, and the initial density is 1×10 8 The adhesion induction rate of the biofilm formed in cells/mL to juvenile mollusk was 27.8%, and the initial density was 1.0X10% 6 The adhesion induction rate of the biofilm formed at the time of cells/mL to juvenile mollusk was 15.6%. The biofilm formed by the bacteria has the activity of inducing the adhesion of the juvenile mollusks of the mytilus coruscus, can be used for inducing the adhesion of the juvenile mollusks of the mytilus coruscus, and can provide a new technical approach for solving the problem of seedling and utilizing bacteria to form the biofilm to induce the adhesion of the juvenile mollusks of the mytilus coruscus.
The previous description of the embodiments is provided to facilitate a person of ordinary skill in the art in order to make and use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments. Those skilled in the art will appreciate that, in light of the principles of the present invention, improvements and modifications can be made without departing from the scope of the invention.
Sequence listing
<110> Shanghai university of ocean
<120> Brevundimonas deep sea and application thereof in inducing adhesion of mytilus coruscus
<141> 2020-11-23
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1338
<212> DNA
<213> Natural seawater ()
<400> 1
acacatgcag tcgaacgaac tcttcggagt tagtggcgga cgggtgagta acacgtggga 60
acgtgccttt aggttcggaa taactcaggg aaacttgtgc taataccgaa tgtgcccttc 120
gggggaaaga tttatcgcct ttagagcggc ccgcgtctga ttagctagtt ggtgaggtaa 180
aggctcacca aggcgacgat cagtagctgg tctgagagga tgatcagcca cattgggact 240
gagacacggc ccaaactcct acgggaggca gcagtgggga atcttgcgca atgggcgaaa 300
gcctgacgca gccatgccgc gtgaatgatg aaggtcttag gattgtaaaa ttctttcagt 360
agggacgata atgacggtac ctacagaaga agccccggct aacttcgtgc cagcagccgc 420
ggtaatacga agggggctag cgttgctcgg aattactggg cgtaaaggga gcgtaggcgg 480
acatttaagt caggggtgaa atcccggggc tcaacctcgg aattgccttt gatactgggt 540
gtcttgagtg tgagagaggt atgtggaact ccgagtgtag aggtgaaatt cgtagatatt 600
cggaagaaca ccagtggcga aggcgacata ctggctcatt actgacgctg aggctcgaaa 660
gcgtggggag caaacaggat tagataccct ggtagtccac gccgtaaacg atgattgcta 720
gttgtcggga tgcatgcatt tcggtgacgc agctaacgca ttaagcaatc cgcctgggga 780
gtacggtcgc aagattaaaa ctcaaaggaa ttgacggggg cccgcacaag cggtggagca 840
tgtggtttaa ttcgaagcaa cgcgcagaac cttaccacct tttgacatgc ctggaccgcc 900
agagagatct ggctttccct tcggggacta ggacacaggt gctgcatggc tgtcgtcagc 960
tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc aaccctcgcc attagttgcc 1020
atcatttagt tgggaactct aatgggactg ccggtgctaa gccggaggaa ggtggggatg 1080
acgtcaagtc ctcatggccc ttacagggtg ggctacacac gtgctacaat ggcgactaca 1140
gagggttaat ccttaaaagt cgtctcagtt cggattgtcc tctgcaactc gagggcatga 1200
agttggaatc gctagtaatc gcggatcagc atgccgcggt gaatacgttc ccgggccttg 1260
tacacaccgc ccgtcacacc atgggagttg gttctacccg aaggcgctgc gctgaccgca 1320
agggggcagg cgaccacg 1338
Claims (4)
1. A strain of Brevundimonas deep sea is characterized by being classified and named as Brevundimonas deep sea（Brevundimonas sp.）c7200a，The Brevundimonas sp.c7200a is preserved in China center for type culture collection, the preservation number is CCTCC NO: M2019802, the preservation date is 10 months 11 days in 2019, and the preservation address is in the eight-path 299 university of Wuhan in Wuhan district of Wuhan, hubei province.
2. The Brevundimonas deep sea according to claim 1, wherein the 16S rRNA gene of Brevundimonas deep sea sp.c7200a is shown in SEQ ID NO. 1.
3. Use of the deep sea shortwave monad of claim 1 or 2 for inducing adhesion of thick shell mussel.
4. The use of claim 3, wherein the mytilus coruscus is juvenile mollusk.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103881947A (en) * | 2014-02-26 | 2014-06-25 | 东北农业大学 | Brevendimonas diminuta strain for forming biofilm and application thereof in cyanide-containing wastewater treatment |
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| ES2477815B1 (en) * | 2013-01-17 | 2015-04-27 | Jorge DÍAZ CRESPO CARDONA | Brine and procedure for its manufacture |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103881947A (en) * | 2014-02-26 | 2014-06-25 | 东北农业大学 | Brevendimonas diminuta strain for forming biofilm and application thereof in cyanide-containing wastewater treatment |
Non-Patent Citations (2)
| Title |
|---|
| Distinctive protein expression patterns of the strain Brevundimonas sp. ZF12 isolated from the aqueous zone containing high levels of radiation to cadmium-induced stress;N. Masoudzadeh et al.;Journal of Biotechnology;第186卷;第49-57页 * |
| 低湿度表面的海洋附着细菌对厚壳贻贝附着的影响;周轩 等;大连海洋大学学报;第30卷(第1期);第30-35页 * |
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