CN104531560A - 一株海洋希瓦氏细菌及其在诱导厚壳贻贝稚贝附着中的应用 - Google Patents
一株海洋希瓦氏细菌及其在诱导厚壳贻贝稚贝附着中的应用 Download PDFInfo
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- 241000490596 Shewanella sp. Species 0.000 title claims abstract description 8
- 230000006698 induction Effects 0.000 title claims abstract description 7
- 241000894006 Bacteria Species 0.000 claims description 24
- 241000237852 Mollusca Species 0.000 claims description 22
- 230000001580 bacterial effect Effects 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 2
- 235000015097 nutrients Nutrition 0.000 claims description 2
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- 238000005406 washing Methods 0.000 claims description 2
- 238000004321 preservation Methods 0.000 abstract description 5
- 238000002474 experimental method Methods 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 description 18
- 235000015170 shellfish Nutrition 0.000 description 4
- 241001029977 Shewanella sp. ECSMB56 Species 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 241000237519 Bivalvia Species 0.000 description 2
- 241000237525 Mytilidae Species 0.000 description 2
- 239000000853 adhesive Substances 0.000 description 2
- 230000001070 adhesive effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 241000143233 Mytilus coruscus Species 0.000 description 1
- 241000237536 Mytilus edulis Species 0.000 description 1
- DPKHZNPWBDQZCN-UHFFFAOYSA-N acridine orange free base Chemical compound C1=CC(N(C)C)=CC2=NC3=CC(N(C)C)=CC=C3C=C21 DPKHZNPWBDQZCN-UHFFFAOYSA-N 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000010062 adhesion mechanism Effects 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- DZBUGLKDJFMEHC-UHFFFAOYSA-N benzoquinolinylidene Natural products C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 230000029052 metamorphosis Effects 0.000 description 1
- 235000020638 mussel Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
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Abstract
本发明公开了一株海洋希瓦氏细菌及其在诱导厚壳贻贝稚贝附着中的应用,保藏编号为CGMCC No:9798,分类命名:希瓦氏菌Shewanella sp。实验证实,本发明成功分离、纯化和鉴定的海洋希瓦氏细菌菌株ECSMB56形成的微生物被膜对厚壳贻贝稚贝附着具有诱导作用。
Description
技术领域
本发明涉及一株海洋希瓦氏细菌及其在诱导厚壳贻贝稚贝附着方面的应用,具体涉及一株海洋希瓦氏细菌(Shewanella sp.ECSMB56)的分离、纯化、鉴定和保藏以及其在不同初始细菌密度条件下形成的微生物被膜对厚壳贻贝稚贝附着的影响作用。
背景技术
因此,现有技术存在缺陷,需要改进厚壳贻贝(Mytilus coruscus),隶属于软体动物门(Mollusca),双壳纲(Bivalvia),贻贝目(Mytilodia),贻贝科(Mytilidae),是我国重要的海产贝类养殖品种,分布于黄海、渤海和东海沿岸。在厚壳贻贝生活史中,浮游生活阶段的幼虫成熟后,便会附着变态成为稚贝,并最终发育为成贝。在由稚贝向成贝生长发育的过程中,当环境发生变化时,厚壳贻贝稚贝能够自行切断足丝,开始爬行并重新选择适宜的附着基进行再次附着。另外,厚壳贻贝具有生长快、抗病能力和环境适应能力强等特点,但由于自然资源的大量采捕和人工培育技术的不足,制约了厚壳贻贝养殖产业的发展。因此,探索利用细菌形成微生物被膜诱导厚壳贻贝附着技术等生物工程技术显得尤为必要。
发明内容
本发明涉及的海洋希瓦氏细菌(Shewanella sp.ECSMB56)来源于浙江省舟山市枸杞岛海域(122°46′E;30°43′N)的自然微生物被膜。利用ZoBell 2216E固体培养基分离、纯化得到纯种菌落,并通过16S rRNA基因测序与GENEBAN(http://www.ncbi.nlm.nih.gov/)中核酸数据进行比对后上传至NCBI数据库。最终确定菌名为希瓦氏细菌,获取序列号为KM369861,该细菌菌株ECSMB56保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编100101,保藏日期为2014年10月20日,保藏编号为CGMCC No:9798,分类命名:希瓦氏菌Shewanella sp.
实验证实,本发明成功分离、纯化和鉴定的海洋希瓦氏细菌菌株ECSMB56形成的微生物被膜对厚壳贻贝稚贝附着具有诱导作用。
附图说明
图1不同初始细菌密度形成的微生物被膜对厚壳贻贝稚贝附着的诱导作用;
具体实施方式
以下结合具体实施例,对本发明进行详细说明。
菌株的分离及保藏:
本发明涉及的海洋希瓦氏细菌(Shewanella sp.ECSMB56)来源于浙江省舟山市枸杞岛海域(122°46′E;30°43′N)的自然微生物被膜。利用ZoBell 2216E固体培养基分离、纯化得到纯种菌落,并通过16S rRNA基因测序与GENEBAN(http://www.ncbi.nlm.nih.gov/)中核酸数据进行比对后上传至NCBI数据库。最终确定菌名为希瓦氏细菌,获取序列号为KM369861,该细菌菌株ECSMB56保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编100101,保藏日期为2014年10月20日,保藏编号为CGMCC No:9798,分类命名:希瓦氏菌Shewanella sp.
本发明涉及的海洋希瓦氏细菌对厚壳贻贝稚贝附着的诱导实验是将纯种菌株先接到ZoBell 2216E液体培养基中扩大培养,离心洗涤并制成悬浮液。然后用吖啶橙染色并在荧光显微镜1000倍镜头下随机选取10个点进行计数,确定菌液总密度为3.4×108cells/ml。之后将一枚灭菌的载玻片放在盛有20ml菌液初始细菌密度为1.0×106cells/ml、3.0×106cells/ml、6.0×106cells/ml和10×106cells/ml的培养皿中,在18℃、黑暗条件下培养48h形成微生物被膜。最后将一枚附有微生物被膜的载玻片及10只厚壳贻贝稚贝(壳长1.22±0.05mm;壳高0.75±0.02mm)放入含有灭菌过滤海水的培养皿中,在12h时记录稚贝的附着率,即载玻片上附着的稚贝个数占该培养皿中稚贝总个数的百分比。
研究发现该细菌形成的微生物被膜对厚壳贻贝稚贝的附着有较高的诱导活性,实验结果如图1所示,在初始密度为10×106cells/ml时(当初始细菌密度大于10×106cells/ml时,附着率基本保持不变),形成的微生物被膜对稚贝附着的诱导率高达55%,相比与无微生物被膜的对照组提高了45%。另外,不同初始密度条件下形成的微生物被膜对稚贝的诱导活性也有所不同,初始密度为1.0×106cells/ml时形成的微生物被膜对稚贝的诱导活性仅为37%。由此可得,该株海洋希瓦氏细菌形成的微生物被膜能够有效诱导海产贝类厚壳贻贝稚贝的附着,本研究成果对于阐明厚壳贻贝稚贝附着机制和厚壳贻贝培育与养殖均具有重要的意义。
应当理解的是,对本领域普通技术人员来说,可以根据上述说明加以改进或变换,而所有这些改进和变换都应属于本发明所附权利要求的保护范围。
Claims (3)
1.一株海洋希瓦氏细菌,保藏编号为CGMCC No:9798,分类命名:希瓦氏菌Shewanella sp.。
2.根据权利要求1所述的海洋希瓦氏细菌在诱导厚壳贻贝稚贝附着中的应用。
3.根据权利要求2所述的应用,其特征在于,将纯种菌株先接种到ZoBell 2216E液体培养基中扩大培养,离心洗涤并制成悬浮液,制得海洋希瓦氏细菌菌液初始细菌密度为1-10×106cells/ml。
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108611298A (zh) * | 2018-05-05 | 2018-10-02 | 上海海洋大学 | 一种深海芽孢杆菌及其在诱导厚壳贻贝稚贝附着中的应用 |
CN110959562A (zh) * | 2019-12-25 | 2020-04-07 | 上海海洋大学 | 牡蛎肽在促进海洋希瓦氏菌诱导厚壳贻贝稚贝附着上的应用 |
CN110982747A (zh) * | 2019-12-25 | 2020-04-10 | 上海海洋大学 | 人工海水组合物、其培养的海洋希瓦氏菌和应用 |
CN112159785A (zh) * | 2020-11-03 | 2021-01-01 | 厦门大学 | 麦氏交替单胞菌e6及其应用 |
Families Citing this family (2)
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CN108531423B (zh) * | 2018-04-08 | 2021-01-26 | 上海海洋大学 | 一种深海盐单胞菌及其在诱导厚壳贻贝稚贝附着中的应用 |
CN108315280B (zh) * | 2018-04-08 | 2021-03-23 | 上海海洋大学 | 一种海洋除烃海杆细菌及其在诱导厚壳贻贝稚贝附着中的应用 |
-
2014
- 2014-12-09 CN CN201410741373.4A patent/CN104531560B/zh active Active
Non-Patent Citations (4)
Title |
---|
GUO,X.P.等: "Shewanella sp. ECSMB56 16S ribosomal RNA gene, partial sequence", 《GENBANK: KM369861.1》 * |
JIN-LONG YANG 等: "Larval settlement and metamorphosis of the mussel Mytilus coruscus in response to monospecific bacterial biofilms", 《BIOFOULING: THE JOURNAL OF BIOADHESION AND BIOFILM》 * |
YI-FENG LI 等: "Effects of bacterial biofilms on settlement of plantigrades of the mussel Mytilus coruscus", 《AQUACULTURE》 * |
杨金龙 等: "微生物膜对厚壳贻贝稚贝附着的影响", 《水产学报》 * |
Cited By (5)
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CN108611298A (zh) * | 2018-05-05 | 2018-10-02 | 上海海洋大学 | 一种深海芽孢杆菌及其在诱导厚壳贻贝稚贝附着中的应用 |
CN108611298B (zh) * | 2018-05-05 | 2021-03-23 | 上海海洋大学 | 一种深海芽孢杆菌及其在诱导厚壳贻贝稚贝附着中的应用 |
CN110959562A (zh) * | 2019-12-25 | 2020-04-07 | 上海海洋大学 | 牡蛎肽在促进海洋希瓦氏菌诱导厚壳贻贝稚贝附着上的应用 |
CN110982747A (zh) * | 2019-12-25 | 2020-04-10 | 上海海洋大学 | 人工海水组合物、其培养的海洋希瓦氏菌和应用 |
CN112159785A (zh) * | 2020-11-03 | 2021-01-01 | 厦门大学 | 麦氏交替单胞菌e6及其应用 |
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