分析化学2016，Vol.44Issue(12)：1867-1873,7.DOI:10.11895/j.issn.0253-3820.160432
三疣梭子蟹肠道菌群结构及代谢表型分析
Gut Microbiota and Metabolic Phenotype of Portunus trituberculatus
摘要
Abstract
16 S rRNA gene high-throughput sequencing and nuclear magnetic resonance ( NMR )-based metabolomics techniques were established to understand the composition of gut microbiota and metabolites of Portunus trituberculatus. Swimming crab gut samples were used for the extraction of bacterial genomic DNA, followed by the amplification of hypervariable domain V3-V4 of 16 S rRNA gene. The obtained PCR products were used for analysis on an Illumina MiSeq platform. The bacterial phylotypes were then clustered and assigned. The results showed that swimming crab gut was dominated by proteobacteria, bacteroidetes, fusobacteria, tenericutes, and acidobacteria at the phylum level. At the genus level, Photobacterium, Paludibacter, and Propionigenium were enriched in it. Crab intestinal samples were extracted with CH3 OH/H2 O ( 2:1, V/V ) solution by shaking with a tissuelyser. After the removal of methanol, the resultant supernatants were lyophilized. Each of lyophilized extract was dissolved into Na+/K+phosphate buffer and then centrifuged at high speed for NMR analysis. A standard noesypr1D was used to acquire 1 H NMR spectra. The 90o pulse length was adjusted to approximately 10 μs. The recycle delay and mixing time were set to 2 s and 100 ms in sequence. The spectral width was set to 20 ppm. Sixty-four transients were collected into 32 k data points for each spectrum. A range of two-dimensional NMR spectra were acquired for the resonance assignment. The results showed that swimming crab gut metabolome comprised 30 metabolites including some amino acids, organic acids, and amines. In summary, this study provided a method for the systematic analysis of the compositions of gut bacterial community and metabolites of swimming crab.关键词
三疣梭子蟹/肠道菌群/高通量测序/代谢表型/核磁共振Key words
Portunus trituberculatus/Gut microbiota/High-throughput sequencing/Metabolic phenotype/Nuclear magnetic resonance引用本文复制引用
曾廷兰,叶央芳,母昌考,王凯,李荣华,王春琳..三疣梭子蟹肠道菌群结构及代谢表型分析[J].分析化学,2016,44(12):1867-1873,7.基金项目
本文系国家自然科学基金(Nos.41673076,41476124),宁波市科技富民计划(No.2016C10062)和宁波大学王宽诚基金资助项目 （Nos.41673076,41476124）
