2种海洋饵料微藻藻际环境的细菌多样性研究
Bacterial Diversity in the Phycosphere of Two Marine Bait-Microalgae
投稿时间:2016-08-09  修订日期:2017-09-13
中文关键词:角毛藻  扁藻  藻际环境  高通量测序
英文关键词:Chaetoceros sp.  Platymonas sp.  phycosphere  high-throughput sequencing
基金项目:浙江省科技厅项目-水产养殖环境微生物种群结构分析及健康养殖微生物系统构建研究(2014C32079);浙江省重点实验室开放基金项目-海洋微藻对多氯联苯的胁迫应答及吸收吸附作用(J2013010)。
作者单位E-mail
黄振华 浙江省海洋水产养殖研究所浙江省近岸水域生物资源开发与保护重点实验室温州 325005 huangzhenhua0112@163.com 
陈星星 浙江省海洋水产养殖研究所浙江省近岸水域生物资源开发与保护重点实验室温州 325005 695495@qq.com 
柯爱英 浙江省海洋水产养殖研究所浙江省近岸水域生物资源开发与保护重点实验室温州 325005 huangzhenhua0112@163.com 
曾国权 浙江省海洋水产养殖研究所浙江省近岸水域生物资源开发与保护重点实验室温州 325005 847771770@qq.com 
吴越 浙江省海洋水产养殖研究所浙江省近岸水域生物资源开发与保护重点实验室温州 325005 476281577@qq.com 
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中文摘要:
      选取对数生长期的海洋饵料微藻——角毛藻(Chaetoceros sp.)和扁藻(Platymonas sp.)藻液,经无菌混合纤维素酯膜抽滤,获得游离和粘附的藻际细菌,并提取基因组DNA进行16SrDNA高通量测序,测序区间为16S rDNA的V3~V4可变区,测序结果经过数据质控获得有效序列数分别为44222和40721,用Uclust v1.1.579对序列进行OTU聚类获得有效OUT数分别为3747和3809;用RDP classifier 进行物种注释,分析藻际环境的细菌多样性。结果表明,2种微藻的藻际细菌主要分布于变形菌门 (Proteobacteria)。角毛藻藻际细菌优势种群主要分布于γ-变形菌纲(34.3%)和α-变形菌纲(16.3%),扁藻藻际细菌优势种群主要分布于γ-变形菌纲(36.9%)、α-变形菌纲(17.7%)和β-变形菌纲(11.4%)。细菌群落Alpha多样性指数表明,角毛藻覆盖率为96.4%,香农指数为6.28,ACE指数为7247.64,Chao1指数为6046.91;扁藻覆盖率为98.0%,香农指数为5.99,ACE指数为7166.06,Chao1指数为6172.99。2藻种的藻际细菌群落内菌种均匀性和多样性相近。
英文摘要:
      Marine bait-microalgae is one of the critical factors affecting development of the aquaculture industry. Marine bait-microalgae is surrounded by a specialized bacterial community called the phycosphere, symbiotic with the marine bait-microalgae and greatly influencing its growth, reproduction and the nutritional value. In this study, we clarified the characteristics of the phycosphere bacterial community for two marine bait-microalgae (Chaetoceros sp. and Platymonas sp.), aiming to promote research on the relationship of bait-microalgae algae and its bacterial community and to provide a reference for the exploitation of marine bait resources. Microalgae suspensions in logarithmic growth phase were filtered through 0.22 μm mixed cellulose ester membranes to collect the microbes. Bacterial DNA was isolated from the membrane filters and high-throughput sequencing (Illumina MiSeq) of the V3–V4 hypervariable region from the 16S rRNA gene was used to investigate microbiome structural characteristics. After ensuring quality control, 44222 filtered sequences for Chaetoceros sp. and 40721 filtered sequences for Platymonas sp. were obtained. Bacterial phylotypes were classified using Uclust v1.1.579 and assigned to operational taxonomic units (OTUs), and the number of efficient OTUs was 3747 for Chaetoceros sp. and 3809 for Platymonas sp.. The taxonomic identity of each phylotype was determined using a RDP classifier and microbial diversity was also analyzed. The dominant OTUs in the phycosphere were Proteobacteria, accounting for 60.81% of the total biomass of the bacterial community in the phycosphere of Chaetoceros sp. and 72.96% for Platymonas sp.. γ-Proteobacteria (34.3%) and α-Proteobacteria (16.3%) were the most dominant classes in the phycosphere of Chaetoceros sp., while ?-Proteobacteria (36.9%), α-Proteobacteria (17.7%) and β-Proteobacteria (11.4%) were the most dominant classes in the phycosphere of Platymonas sp.. The microbial community coverage in the phycospheres of Chaetoceros sp. and Platymonas sp. were 96.4% and 98.0%, both with high species richness. The species number and bacterial genus of the two bacterial communities were similar, with 10 bacterial genera in common, 3 peculiar bacterial genera for Chaetoceros sp. and 2 peculiar bacterial genera for Platymonas sp.. Additionally, the Shannon, ACE and Chao1 indices of the bacterial communities in the phycospheres of Chaetoceros sp. and Platymonas sp. were, respectively, 6.28, 7247.64, 6046.91 and 5.99, 7166.06, 6172.99, indicating that the phycosphere bacterial communities of the two marine bait-microalgae are similar in uniformity and diversity.
黄振华,陈星星,柯爱英,曾国权,吴越.2018.2种海洋饵料微藻藻际环境的细菌多样性研究[J].水生态学杂志,39(1):44-49.
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