生物技术通报 ›› 2020, Vol. 36 ›› Issue (11): 76-84.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0187
收稿日期:
2020-02-27
出版日期:
2020-11-26
发布日期:
2020-11-20
作者简介:
刘倩倩,女,硕士研究生,研究方向:海洋污损生物菌群;E-mail: 基金资助:
LIU Qian-qian(), SHI Hong-wei, GUO Chang-lu, ZHANG Zhi-zhou()
Received:
2020-02-27
Published:
2020-11-26
Online:
2020-11-20
摘要:
本研究利用环氧铁红防锈漆和丙烯酸脂成膜物作为涂层基质,添加4种纳米材料(0.1%,W/V)制备了2类防污涂层,发现两类涂层的防污性能打分接近,但是基于环氧铁红防锈漆的涂层其玻璃海鞘附着程度显著高于丙烯酸酯。通过实海挂板采集涂层表面生物膜,提取其宏基因组DNA并进行16S核糖体RNA全长基因的高通量测序,研究了第28天涂层表面原核菌群的组成差异。结果显示:(1)丙烯酸酯涂层生物膜原核菌群多样性高于铁红涂层;(2)纳米材料的添加没有明显改变涂层表面菌群结构组成;(3)两类涂层表面菌群结构差异明显,尤其是Neptuniibacter sp. CAR-SF和Cellvibrionales unclassified菌的含量在铁红涂层中含量均在4%左右,而在丙烯酸涂层中几乎为零,对应的OTU序列读数在两类涂层中相差100倍以上,差异性明显;而qPCR结果进一步证实Neptuniibacter属在两类涂层中含量相差很大。基于丙烯酸酯基质的涂层其海鞘附着量明显减少是否与上述细菌含量差异相关值得进一步研究。
刘倩倩, 史宏伟, 郭长禄, 张治洲. 海鞘附着相关微生物膜中细菌原核群落结构解析[J]. 生物技术通报, 2020, 36(11): 76-84.
LIU Qian-qian, SHI Hong-wei, GUO Chang-lu, ZHANG Zhi-zhou. Composition Analysis of Prokaryotic Flora in the Marine Biofilm Correlated with Ascidian Settlement[J]. Biotechnology Bulletin, 2020, 36(11): 76-84.
编号 | 材料 | 外径 | 长度或颗粒大小 | 纯度 |
---|---|---|---|---|
D19 | Multi-wall Carbon Nanotubes | 10-20 nm | 30-100 μm | >95% |
D33 | Carboxyl Multi-wall Carbon Nanotubes(-COOH) | 10-20 nm | 10-30 μm | >95% |
D34 | Carboxyl Multi-wall Carbon Nanotubes(-COOH) | 20-30 nm | ~30 μm | >95% |
DPt | Platinum nanopowder(CAS:7440-06-4) | —— | 200 nm | 99.9% |
表1 纳米材料信息
编号 | 材料 | 外径 | 长度或颗粒大小 | 纯度 |
---|---|---|---|---|
D19 | Multi-wall Carbon Nanotubes | 10-20 nm | 30-100 μm | >95% |
D33 | Carboxyl Multi-wall Carbon Nanotubes(-COOH) | 10-20 nm | 10-30 μm | >95% |
D34 | Carboxyl Multi-wall Carbon Nanotubes(-COOH) | 20-30 nm | ~30 μm | >95% |
DPt | Platinum nanopowder(CAS:7440-06-4) | —— | 200 nm | 99.9% |
涂层 | 高通量 测序分组 | 海鞘面积占比/% | 海鞘面积 平均占比 |
---|---|---|---|
铁红/D19 | S4 | 0/3.97/8.02/6.77 | 4.69% |
铁红/D33 | 0.98/0.89/5.64/5.01 | 3.13% | |
铁红/D34 | 0.5/3.65/7.38/7.63 | 4.79% | |
铁红/DPt | 3.89/4.68/6.08/7.23 | 5.47% | |
铁红 | S3 | 3.51/1.03/0.67/4.15 | 2.34% |
丙烯酸/D19 | S8 | 0/0/0/0 | 0 |
丙烯酸/D33 | 0/0/0/0 | 0 | |
丙烯酸/D34 | 0/0/0/0 | 0 | |
丙烯酸/DPt | 0/0/0/0 | 0 | |
丙烯酸 | S7 | 0/0/0/0 | 0 |
表2 第28天观察结果
涂层 | 高通量 测序分组 | 海鞘面积占比/% | 海鞘面积 平均占比 |
---|---|---|---|
铁红/D19 | S4 | 0/3.97/8.02/6.77 | 4.69% |
铁红/D33 | 0.98/0.89/5.64/5.01 | 3.13% | |
铁红/D34 | 0.5/3.65/7.38/7.63 | 4.79% | |
铁红/DPt | 3.89/4.68/6.08/7.23 | 5.47% | |
铁红 | S3 | 3.51/1.03/0.67/4.15 | 2.34% |
丙烯酸/D19 | S8 | 0/0/0/0 | 0 |
丙烯酸/D33 | 0/0/0/0 | 0 | |
丙烯酸/D34 | 0/0/0/0 | 0 | |
丙烯酸/DPt | 0/0/0/0 | 0 | |
丙烯酸 | S7 | 0/0/0/0 | 0 |
ace | chao | coverage | shannon | simpson | observed_otus | |
---|---|---|---|---|---|---|
S3 | 844 | 677 | 0.966615 | 3.93 | 0.0689 | 461 |
S4 | 721 | 693 | 0.981109 | 3.85 | 0.0605 | 384 |
S7 | 753 | 656 | 0.967809 | 4.22 | 0.0492 | 422 |
S8 | 610 | 579 | 0.974431 | 4.27 | 0.0428 | 413 |
表3 原核菌落多样性参数
ace | chao | coverage | shannon | simpson | observed_otus | |
---|---|---|---|---|---|---|
S3 | 844 | 677 | 0.966615 | 3.93 | 0.0689 | 461 |
S4 | 721 | 693 | 0.981109 | 3.85 | 0.0605 | 384 |
S7 | 753 | 656 | 0.967809 | 4.22 | 0.0492 | 422 |
S8 | 610 | 579 | 0.974431 | 4.27 | 0.0428 | 413 |
涂层 测试液体 | 铁红 | 铁红/D19 | 铁红/D33 | 铁红/D34 | 铁红/DPt | 丙烯酸 | 丙烯酸/D19 | 丙烯酸/D33 | 丙烯酸/D34 | 丙烯酸/DPt |
---|---|---|---|---|---|---|---|---|---|---|
纯水 | 90 | 83 | 86 | 82 | 79 | 82 | 79 | 79 | 83 | 84 |
海水 | 85 | 78 | 81 | 82 | 75 | 86 | 84 | 83 | 85 | 87 |
表4 全部涂层的静态接触角
涂层 测试液体 | 铁红 | 铁红/D19 | 铁红/D33 | 铁红/D34 | 铁红/DPt | 丙烯酸 | 丙烯酸/D19 | 丙烯酸/D33 | 丙烯酸/D34 | 丙烯酸/DPt |
---|---|---|---|---|---|---|---|---|---|---|
纯水 | 90 | 83 | 86 | 82 | 79 | 82 | 79 | 79 | 83 | 84 |
海水 | 85 | 78 | 81 | 82 | 75 | 86 | 84 | 83 | 85 | 87 |
[1] | 严涛, 韩帅帅, 王建军, 等. 污损性海鞘的生态特点研究展望[J]. 生态学报, 2017,37(20):6647-6655. |
Yan T, Han SS, Wang JJ, et al. An overview of fouling ascidian[J]. Acta Ecologica Sinica, 2017,37(20):6647-6655. | |
[2] | 杨金龙, 王冲, 顾忠旗, 等. 微生物膜对海洋无脊椎动物幼体附着变态的影响研究[J]. 海洋科学, 2012,36(8):116-121. |
Yang JL, Wang C, Gu ZQ, et al. A review on the role of marine biofilms on larval settlement and metamorphosis of marine invertebrates[J]. Marine Sciences, 2012,36(8):116-121. | |
[3] |
Lau SCK, Thiyagarajan V, Cheung SCK, et al. Roles of bacterial community composition in biofilms as a mediator for larval settlement of three marine invertebrates[J]. Aquatic Microbial Ecology, 2005,38(1):41-51.
doi: 10.3354/ame038041 URL |
[4] | Huang SY, Hadfield MG. Composition and density of bacterial biofilms determine larval settlement of the polychaete Hydroides elegans[J]. Marine Ecology Progress Series, 2003,260:161-172. |
[5] | Aldred N, Hoeg JT, Maruzzo D, et al. Analysis of the behaviours mediating barnacle cyprid reversible adhesion[J]. PLoS One, 2013,8:10. |
[6] | Petrone L, Kumar A, Sutanto CN, et al. Mussel adhesion is dictated by time-regulated secretion and molecular conformation of mussel adhesive proteins[J]. Nature Communications, 2015,6:12. |
[7] | Gohad NV, Aldre N, Hartshorn CM, et al. Synergistic roles for lipids and proteins in the permanent adhesive of barnacle larvae[J]. Nature Communications, 2014,5:9. |
[8] |
Misic C, Harrlague AC. Development of marine biofilm on plastic:ecological features in different seasons, temperatures, and light regimes[J]. Hydrobiologia, 2019,835:129-145.
doi: 10.1007/s10750-019-3934-7 URL |
[9] | Olivier F, Tremblay R, Bourget E, et al. Barnacle settlement:field experiments on the influence of larval supply, tidal level, biofilm quality and age on Balanus amphitrite cyprids[J]. Marine Ecology Progress Series, 2000,199:185-204. |
[10] | Szewzyk U, Holmstrom C, Wrangstadh M, et al. Relevance of the exopolysaccharide of marine Pseudomonas sp. Strain S9 for the attachment of Ciona intestinalis larvae[J]. Marine Ecology Progress Series, 1991,75(2-3):259-265. |
[11] |
Holmstrom C, Rittschof D, Kjelleberg S. Inhibition of settlement by larvae of Balanus amphitrite and Ciona intestinalis by a surface-colonizing marine bacterium[J]. Applied and Environmental Microbiology, 1992,58(7):2111-5.
URL pmid: 16348728 |
[12] | Keough MJ, Raimondi PT. Responses of settling invertebrate larvae to bioorganic films:effects of different types of films[J]. Journal of Experimental Marine Biology and Ecology, 1995,185:235-253. |
[13] | Holmstrom C, Rittschof D, Kjelleberg S, et al. Inhibition and facilitation of bryozoan and ascidian settlement by natural multispecies biofilms:effects of film age and the roles of active and passive larval attachment[J]. Marine Biology, 1997,128:463-473. |
[14] | 张朝霞, 柯才焕, 冯丹青, 等. 海洋附着细菌对冠瘤海鞘幼体附着和变态的影响[J]. 海洋学报, 2005(5):96-102. |
Zhang ZX, Ke CH, Feng DQ, et al. Influences of marine adhesive bacteria on settlement and metamorphosis of Styela conopus Savigny larvae[J]. Haiyang Xuebao, 2005(5):96-102. | |
[15] |
Zardus JD, Nedved BT, Huang Y, et al. Microbial biofilms facilitate adhesion in biofouling invertebrates[J]. Biological Bulletin, 2008,214:91-98.
doi: 10.2307/25066663 URL pmid: 18258779 |
[16] | 孙源. CNTs/PDMS复合涂层对污损早期微生物群落影响研究[D]. 威海:哈尔滨工业大学, 2016. |
Sun Y. The study on the effect of the CNTs/PDMS composites on the pioneer fouling microbial communities[D]. Weihai: Harbin Institute of Technology, 2016 | |
[17] | Sun Y, Zhang ZZ. New anti-biofouling carbon nanotubes-filled polydimethylsiloxane composites against colonization by pioneer eukaryotic microbes[J]. International Biodeterioration & Biodegradation, 2016,110, 147-154. |
[18] | 刘倩倩, 李俊薇, 曹润洁, 等. 耐酸乳酸杆菌物种特异性引物设计及其在古井贡酒窖泥酒醅质量评价中的初步应用[J]. 食品与发酵工业, 2019,45(6):16-22. |
Liu QQ, Li JW, Cao RJ, et al. Employment of Primer-Blast to design speciesspecific primers for Lactobacillus acetotolerans and their implications in quality assessment of pitmud or Zaopei samples in Gujing Tribute liquor[J]. Food and Fermentation Industries, 2019,45(6):16-22. | |
[19] | 栾鑫, 段继周, 陈永伟, 等. 两种船用防污涂层实海挂片表面细菌群落多样性分析[J]. 应用与环境生物学报, 2013,19(3):471-477. |
Luan X, Duan JZ, Chen YW, et al. Diversity of bacterial community on the surface of marine paint in diversity of bacterial community on the surface of marine paint in temperate coastal marine waters temperate coastal marine waters[J]. Chinese Journal of Applied and Environmental Biology, 2013,19(3):471-477. | |
[20] |
Khandavalli LVNS, Lodha T, Abdullah M, et al. Insights into the carbonic anhydrases and autotrophic carbon dioxide fixation pathways of high CO2 tolerant Rhodovulum viride JA756[J]. Microbiological Research, 2018,215, 130-140.
doi: 10.1016/j.micres.2018.07.006 URL pmid: 30172299 |
[21] | 胡晓娟, 文国樑, 李卓佳, 等. 养殖中后期高位池对虾水体微生物群落结构及水体理化因子[J]. 生态学杂志, 2018,37(1):171-178. |
Hu XJ, Wen GL, Li, ZJ, et al. Microbial community structure and physicochemical factors in the water of upland pond of Litopenaeus vannamei during the middle and late stage of culture[J]. Chinese Journal of Ecology, 2018,37(1):171-178. | |
[22] |
Nagashima H, Bin Zulkharnain A, Maeda R, et al. Cloning and nucleotide sequences of carbazole degradation genes from marine bacterium Neptuniibacter sp strain CAR-SF[J]. Current Microbiology, 2010,61:50-56.
doi: 10.1007/s00284-009-9575-8 URL pmid: 20039169 |
[23] | Dieguez AL, Romalde JL. Draft genome sequences of Neptuniibacter sp. strains LFT 1. 8 and ATR 1. 1[J]. Microbiology Resource Announcements, 2017,5:2. |
[24] |
Beigbeder A, Degee P, Conlan SL, et al. Preparation and characterization of silicone-based coatings filled with carbon nanotubes and natural sepiolite and their application as marine fouling-release coatings[J]. Biofouling, 2008,24(4), 291-302.
URL pmid: 18568667 |
[25] |
Beigbeder A, Mincheva R, Pettitt ME, et al. Marine fouling release silicone/carbon nanotube nanocomposite coatings:on the importance of the nanotube dispersion state[J]. Journal of Nanoscience and Nanotechnology, 2010,10:2972-2978.
URL pmid: 20358887 |
[26] | 邓冰锋, 谢志鹏, 王晶晶, 等. 丙烯酸锌自抛光防污涂料的配套性及防污性能研究[J]. 材料开发与应用, 2017,32(1):12-17. |
Deng BF, Xie ZP, Wang JJ, et al. Research on matching and antifouling performance of Zinc acrylate self-polishing antifouling Paint[J]. Development and Application of Materials, 2017,32(1):12-17. |
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[9] | 石玉杰;姚洪军;. 生物膜脂及脂肪酸的分析方法评述[J]. , 2007, 0(05): 89-92. |
[10] | . 国外动态[J]. , 2003, 0(01): 16-22. |
[11] | . 环境保护及农业废物利用[J]. , 1996, 0(05): 106-107. |
[12] | . 环境保护及农业废物利用[J]. , 1995, 0(06): 104-108. |
[13] | 朱遐. 来自海洋的工业产品和药物[J]. , 1995, 0(05): 29-30. |
[14] | . 环境保护及农业废物利用[J]. , 1994, 0(03): 109-114. |
[15] | . 环境保护及农业废物利用[J]. , 1993, 0(11): 88-98. |
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