海鞘附着相关微生物膜中细菌原核群落结构解析

doi:10.13560/j.cnki.biotech.bull.1985.2020-0187

摘要/Abstract

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 28^th 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

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.

图/表 10

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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%

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

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