生物技术通报 ›› 2020, Vol. 36 ›› Issue (11): 76-84.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0187

• 研究报告 • 上一篇    下一篇

海鞘附着相关微生物膜中细菌原核群落结构解析

刘倩倩(), 史宏伟, 郭长禄, 张治洲()   

  1. 1.哈尔滨工业大学(威海)海洋科学与技术学院,威海 264209
    2.山东省海洋船舶防污工程技术研究中心,威海 264209
  • 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: 592181869@qq.com
  • 基金资助:
    国家自然科学基金项目(31071170);山东省重点研发计划(2016GSF115022);山东省自然科学基金项目(ZR2018MC002);威海市科技创新项目(WH2018019)

Composition Analysis of Prokaryotic Flora in the Marine Biofilm Correlated with Ascidian Settlement

LIU Qian-qian(), SHI Hong-wei, GUO Chang-lu, ZHANG Zhi-zhou()   

  1. 1. School of Marine Science and Technology,Harbin Institute of Technology,Weihai 264209
    2. Marine Antifouling Engineering Technology Center of Shandong Province,Weihai 264209
  • 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。

关键词: 生物膜, 铁红, 丙烯酸酯, 原核菌群多样性, 海鞘

Abstract:

Using iron oxide red paint and acrylate as coating base,4 different nanomaterials(0.1%,W/V)were added to make 2 types of antifouling coatings. The two types of coating performed similarly in the context of antifouling scores,but significantly more Ciona intestinalis attached on the iron oxide red paint coatings than acrylate ones. Biofilm samples were collected from marine hanging plates,the metagenome DNA samples from the 28th day biofilm were subjected to high-throughput sequencing of full-length 16S rRNA,and the composition differences of prokaryotic flora between 2 coatings were analyzed. The results indicated that:1)the alpha diversity of prokaryotic flora in the acrylate coating was higher than that in the iron oxide red paint coating;2)interestingly,nanomaterials didn’t result in the significant composition changes in the prokaryotic flora on the both coatings;and 3)there were significant differences in the abundance of several bacterial strains between two types of coatings,especially for two dominant bacteria Neptuniibacter sp. CAR-SF and Cellvibrionales unclassified at 4% on the iron oxide red paint coating but almost zero on the acrylate coating. The observed OTU numbers of the two bacteria on 2 coatings were of over 100-fold difference,and the qPCR results confirmed that the contents of Neptuniibacter genus on 2 coatings differed remarkably. Whether the decrease in the amount of ascidian attached to the acrylate-based coating is related to the above bacterial content difference is worthy of further study.

Key words: biofilm, iron oxide red paint, acrylate, diversity of prokaryotic flora, ascidian