蟋蟀肠道细菌多样性的宏基因组分析
Metagenome analysis of bacterial diversity in cricket intestines
张 科1** 刘伍限2 郑新华1 刘冰许3
点击：1506次 下载：9次
DOI：10.7679/j.issn.2095-1353.2017.050
作者单位：1. 平顶山学院，平顶山 467000；2. 河南省郑州市动物卫生监督所，郑州 450000； 3. 郑州市动物园，郑州 450000
中文关键词： 油葫芦属，MtDNA-16S rRNA，肠道细菌，多样性，16S rRNA
英文关键词：Teleogryllus, MtDNA-16S rRNA, intestine bacteria, bacterial diversity, 16S rRNA
中文摘要：
【目的】 了解油葫芦属蟋蟀成虫前中肠和后肠肠道细菌的多样性。【方法】 首先，基于mtDNA-16S rRNA基因对蟋蟀进行分子鉴定，然后从蟋蟀前中肠和后肠分别提取总DNA，再使用Illumina Miseq平台进行细菌16S rRNA v3-v4变异区进行测序，最后分析蟋蟀肠道微生物的群落组成和多样性。【结果】 蟋蟀mtDNA-16S rRNA基因分析结果显示归属于油葫芦属，暂时命名为平顶山油葫芦。细菌16S rRNA基因宏基因组测序分析，前中肠共获得21 799条reads，857个操作分类单元（Operational taxonomic unit，OUT），后肠获得16 515条reads，2 155个OUT。99%的蟋蟀肠道细菌归属于变形菌门（Proteobacteria）、厚壁菌门（Firmicutes）和拟杆菌门（Bacteroidetes）。前中肠归入232个属，前5个属分别是：乳球菌属（33.47%），泛菌属（21.50%），肠杆菌属（15.47%），沃尔巴克体属（9.47%）和魏斯菌属（5.34%）；后肠归入152个属，前5个属分别是：未分类的瘤胃球菌科成员(Unclassified Ruminococcaceae)（30.07%），Parabacteroides（10.93%），Incertae sedis（9.74%），Alistipes（5.86%），类杆菌属（5.50%），Dysgonomonas（4.91%）。【结论】 mtDNA-16S rRNA是油葫芦属分子鉴定的有效工具。蟋蟀肠道含有丰富的微生物资源，为后续的肠道微生物资源开发奠定了基础。
英文摘要：
[Objectives] To clarify bacterial diversity in the anterior midgut and posterior intestine of Crickets. [Methods] Molecular identification of crickets was carried out based on the cricket 16S rRNA sequence. Total DNA from the anterior midgut and posterior intestine was then isolated and bacterial 16S rRNA v3-v4 sequenced by Illumina Miseq. Finally, the bacterial community composition and diversity in anterior midgut and posterior intestine was analyzed. [Results] Analysis of MtDNA-16S rRNA showed that all crickets belonged to the genus Teleogryllus, with the provisional species name Teleogryllus spp. from Pingdingshan. A total of 21 799 reads clustered into 875 OUTs were obtained from the anterior midgut, and 16 515 reads clustered into 2 155 OUTs from the posterior intestine. Proteobacteria, Firmicutes and Bacteroidetes accounted for 99% of the bacteria identified in both the anterior midgut and posterior intestine. At the genus level, Lactococcus (33.47%), Pantoea (21.50%), Enterobacter (15.47%), Wolbachia (9.47%)and Weissella (5.34%) were the most abundant of 232 genera identified in the anterior midgut, and unclassified Ruminococcaceae (30.07%), Parabacteroides (10.93%), Incertae sedis (9.74%), Alistipes (5.86%), Bacteroides (5.50%), Dysgonomonas (4.91%) were the most abundant of 152 genera identified in the posterior intestine. [Conclusion] MtDNA-16S rRNA is an effective tool for Teleogryllus molecular identification. Cricket intestines are rich in microbial resources and may have the potential for the development of such resources.