基于高通量测序的不同养殖系统下凡纳滨对虾肠道和水体中微生物的多样性

Analysis of Aquaculture-water and Gut from Fenneropenaeus chinensis in Different Systems on Bacterial Diversity based on High-throughput Sequencing Technology

关键词:  高通量测序  凡纳滨对虾  肠道菌群  微生物多样性

Key words:  high-throughput sequencing  Fenneropenaeus chinensis  gut microorganism  bacterial diversity

摘要:

[目的]研究不同养殖系统对凡纳滨对虾肠道及水体微生物多样性的影响。[方法]以循环养殖系统和非养殖系统下凡纳滨对虾养殖水体及其肠道为研究对象,在养殖30 d后采集样品,采用Illumina MiSeq高通量测序样品细菌16S rRNA基因V3+V4区,通过RDP11.3、Greengenes13.8、NCBI 16S Microbial和Customized batabase等数据库进行分析,阐述循环养殖系统的意义和微生物在其中的作用。[结果]养殖期间,不同养殖系统凡纳滨对虾均正常存活。将原始序列优化后对Clean序列进行分析,非循环系统下水体优质序列百分比约81%,肠道中约89%;循环系统下水体优质序列百分比为92%,肠道中为94%。两种养殖系统下水体和肠道细菌在纲水平占绝对优势的是变形菌纲,但各菌群丰度不同;不同的Alpha指数显示出物种间的差异性;通过系统发育树可以看出,不同养殖系统改变了凡纳滨对虾生境中的物种及发育关系,间接阐述微生物在养殖生境中的作用和意义。[结论]通过高通量测序揭示不同养殖系统中微生物菌群的差异以及多样性,对于凡纳滨对虾实际养殖具有指导作用,对阐述养殖过程中微生物和相关疾病的关系具有重要意义。

Abstract:

[Objective] To study the effect of different aquaculture system on bacterial diversity of water and gut from Fenneropenaeus chinensis.[Methods] Taking aquaculture-water and gut of Fenneropenaeus chinensis under both the recirculation system and the non-culture system as the research object,samples were collected after fed 30 d.The bacterial 16S rRNA gene V3+V4 region was sequenced using the Illumina MiSeq high-throughput sequencing sample and analyzed by RDP11.3,Greengenes13.8,NCBI16S Microbial and Customized Database to elucidate the significance of circulatory aquaculture system and the role of microorganisms.[Results] Fenneropenaeus chinensis were survived normally in different culture systems during the cultivation.After the original sequence was optimized,the Clean sequence was analyzed.The percentage of high quality sequences in the non-circulatory system was about 81% and about 89% in the gut.The percentage of high quality sequence in the circulatory system was 92% and 94% in the gut.The Proteobacteria of the water bodies and gut bacteria in the two aquaculture systems were at an absolute advantage,but the abundance of each group was different.The different Alpha-varieties showed the differences among the species.According to the phylogenetic tree,different culture systems had changed the species and developmental relationships of Fenneropenaeus chinensis in habitats and indirectly explained the role and significance of microorganisms in culture habitats.[Conclusion] The high-throughput sequencing reveals differences and diversity of microbial flora in different culture systems and is instructive to the actual culture of Fenneropenaeus chinensis, which is of great significance for elucidating the relationship between microorganisms and related diseases in the aquaculture.