广粪蚊不同发育阶段体内细菌多样性研究

Diversity Analysis of Bacteria in Cobolidia fuscipes (Meigen) at Different Development Stages (Diptera: Scastopsidae)

摘要

利用Illumina MiSeq高通量测序技术, 对广粪蚊不同发育阶段体内细菌16S rDNA的V3~V4变异区进行分析, 探究其细菌的群落多样性。7个样本共获得548 058条有效序列, 它们聚类为870个操作分类单元(operational taxonomic unit, OTU), 注释到24门、51纲、92目、175科、346属。广粪蚊体内细菌的赵氏指数(Chao index)、艾斯指数(Ace index)和香农指数(Shannon index)分别为485.00~885.03、485.09~877.55和2.90~4.31。其中, 优势菌门为变形菌门(Proteobacteria)和放线菌门(Actinobacteria), 优势菌目为根瘤菌目(Rhizobiales)和放线菌目(Actinomycetales), 优势菌属为黄色杆菌科未知属(unclassified Xanthobacteraceae)、假单胞菌属(Pseudo-monas)和无色杆菌属(Leucobacter)。但各样本的优势菌属有所不同, 如假单胞菌属在卵中的占比最高, 黄色杆菌科未知属在1~3龄幼虫和成虫中的相对丰度最高, 棒状杆菌属(Corynebacterium)在4龄幼虫中的占比最高, 普罗威登斯菌属(Providencia)在蛹中的占比最高。主成分分析(principle component analysis, PCA)表明, 广粪蚊体内细菌的群落组成在不同发育阶段存在一定的差异。研究结果为广粪蚊体内微生物资源的开发提供了相关信息, 也为广粪蚊的生物防治工作提供了参考依据。

Abstract

To explore the diversity of bacterial community in Cobolidia fuscipes (Meigen), the V3~V4 regions of bacterial 16S rDNA in this insect species at different development stages were sequenced using Illumina MiSeq high-throughput sequencing platform. A total of 548 058 reads and 870 operational taxonomic units (OTUs) were obtained and annotated into 24 phyla, 51 classes, 92 orders, 175 families and 346 genera. The Chao, Ace and Shannon indices were 485.00~885.03, 485.09~877.55 and 2.90~4.31, respectively. The domi-nant bacterial phyla were Proteobacteria and Actinobacteria, dominant orders were Rhizobiales and Actino-mycetales, and dominant genera were unclassified Xanthobacteraceae, Pseudomonas and Leucobacter. However, the dominant genus in each sample was different. Pseudomonas occupied the highest proportion in eggs, unclas-sified Xanthobacteraceae had the highest relative abundance in the 1st to 3rd instar larvae and adults, Cory-nebacterium had the highest relative abundance in the 4th instar larvae, and Providencia had the highest rela-tive abundance in pupae. Principle component analysis (PCA) demonstrated that the composition of bacterial community in C. fuscipes (Meigen) was different at different development stages. This study provides a theoreti-cal basis for exploitation of microbial resources in C. fuscipes (Meigen) and also for biocontrol of the pest species.

关键词

广粪蚊 / 高通量测序 / 体内细菌 / 多样性

Key words

Cobolidia fuscipes (Meigen) / high-throughput sequencing / bacterium / diversity