昆虫学报 ›› 2023, Vol. 66 ›› Issue (10): 1374-1384.doi: 10.16380/j.kcxb.2023.10.011

• 研究论文 • 上一篇    下一篇

纯水培养对重金属污染地区红裸须摇蚊幼虫肠道细菌群落多样性及潜在功能的影响

马微微, 徐海轩, 曹威, 闫春财, 孙泽阳*   

  1. (天津师范大学生命科学学院, 天津市动物多样性保护与利用重点实验室, 天津 300387)
  • 出版日期:2023-10-20 发布日期:2023-11-27

Effects of pure water culture on the diversity and potential function of bacterial communities in the larval gut of Propsilocerus akamusi (Diptera: Chironomidae) in heavy metal polluted areas

MA Wei-Wei, XU Hai-Xuan, CAO Wei, YAN Chun-Cai, SUN Ze-Yang*   

  1.  (Tianjin Key Laboratory of Conservation and Utilization of Animal Diversity, College of Life Sciences, Tianjin Normal University, Tianjin 300387, China)
  • Online:2023-10-20 Published:2023-11-27

摘要: 【目的】通过对在重金属污染的天津市渤海湾淡水区域鉴定出的摇蚊耐污种红裸须摇蚊Propsilocerus akamusi进行肠道细菌群落多样性和潜在功能研究,了解生境的变化对摇蚊幼虫肠道细菌群落的影响。【方法】将在受重金属污染的天津市渤海湾淡水区域鉴定到的红裸须摇蚊4龄幼虫用实验室蒸馏水饲养7 d作为实验室培养组,并提取实验室培养组和野外采集组的红裸须摇蚊4龄幼虫肠道细菌基因组DNA,对16S rRNA基因V3-V4区进行高通量测序,将测序结果进行数据质控、序列对比和过滤,分析肠道细菌群落物种组成变化并预测肠道细菌潜在功能。【结果】基于红裸须摇蚊4龄幼虫肠道细菌16S rRNA测序结果共注释到11个门、13个纲、33个目、54个科、71个属、90个种以及105个可操作分类单元(operational taxonomic units, OTUs)。实验室培养组红裸须摇蚊4龄幼虫肠道内细菌群落多样性和丰度较野外采集组下降,两组优势菌门相似,均包含厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、拟杆菌门(Bacteroidota)及脱硫弧菌门(Desulfobacterota),野外采集组4龄幼虫肠道内变形菌门丰度显著高于实验室培养组的;实验室培养组4龄幼虫肠道内气单胞菌属Aeromonas、希瓦氏菌属Shewanella、沙雷氏菌属Serratia、假单胞菌属Pseudomonas以及耶尔森氏菌属Yersinia的平均丰度较野外采集组显著降低。线性判别分析结果揭示,野外采集组和实验室培养组的红裸须摇蚊4龄幼虫肠道内存在丰度差异显著的细菌物种。KEGG分析结果显示,红裸须摇蚊4龄幼虫肠道内细菌基因组中有关新陈代谢的基因相对丰度极高,实验室培养组红裸须摇蚊4龄幼虫肠道内细菌基因组中有关环境信息处理和细胞进程的基因相对丰度较野外采集组的显著下降。【结论】本研究结果表明,野外不良环境中生存的红裸须摇蚊幼虫与实验室纯水环境培养幼虫肠道细菌群落多样性以及基因功能存在显著差异,这有助于从环境角度切入研究摇蚊个体抗性机制,为进一步探索摇蚊幼虫肠道内共生微生物抵抗环境胁迫发挥必要作用的研究提供新思路,也为昆虫在不良环境中的耐受机制与其肠道菌群稳态调控机理的研究奠定了基础。

关键词: 红裸须摇蚊, 肠道细菌, 污染, 多样性, 功能预测

Abstract: 【Aim】To understand the effects of habitat changes on the bacterial communities in the larval gut of chironomids by studying the diversity and potential function of bacterial communities in the gut of Propsilocerus akamusi, a pollution-resistant chironomid identified in the freshwater area of the heavy metal polluted Bohai Bay of Tianjin City. 【Methods】The 4th instar larvae of P. akamusi identified in the freshwater area of the heavy metal polluted Bohai Bay of Tianjin City were raised with distilled water in laboratory for 7 d as the laboratory-cultured group, and the bacterial genomic DNA in the 4th instar larval gut of P. akamusi from the laboratory-cultured group and the wild-captured group was extracted. The high-throughput sequencing of the V3-V4 region of 16S rRNA gene was carried out, and the sequencing results were subjected to data quality control, sequence alignment and filtering, the changes in the species composition of gut bacterial communities were analyzed and the potential functions of gut bacteria were predicted. 【Results】Based on the 16S rRNA sequencing results of the gut bacteria in the 4th instar larvae of P. akamusi, 11 phyla, 13 classes, 33 orders, 54 families, 71 genera, 90 species and 105 operational taxonomic units (OTUs) were annotated. The diversity and abundance of bacterial communities in the 4th instar larval gut of P. akamusi in the laboratory-cultured group were lower than those in the wild-captured group. The dominant bacterial phyla in the 4th instar larval gut of the two groups were similar, including Firmicutes, Proteobacteria, Bacteroidota and Desulfobacterota. The abundance of Proteobacteria in the 4th instar larval gut of the wild-captured group was significantly higher than that in the laboratory-cultured group. The average abundance of Aeromonas, Shewanella, Serratia, Pseudomonas and Yersinia in the 4th instar larval gut in the laboratory-cultured group was significantly lower than that in the wild-captured group. The results of linear discriminant analysis revealed that there were bacterial species with significantly different abundance in the 4th instar larval gut of P. akamusi between the wild-captured group and the laboratory-cultured group. The KEGG analysis results showed that the relative abundance of metabolism-related genes in the bacterial genome of the 4th instar larval gut of P. akamusi was extremely high. The relative abundance of genes related to environmental information processing and cellular processes in the gut bacterial genome of the 4th instar larva of P. akamusi in the laboratory-cultured group significantly decreased as compared to that in the wild-captured group.【Conclusion】The results of this study indicate that there are significant differences in the diversity of the gut bacterial communities and gene functions between P. akamusi larvae living in adverse field environments and those reared in laboratory pure water environments. This helps to study the individual resistance mechanisms of chironomids from an environmental perspective, provides a new idea for further exploring the mechanism of the symbiotic microorganisms in the gut of chironomid larvae to cope with environmental stress, and also lays a foundation for the study of the tolerance mechanism of insects in adverse environmental conditions and the regulatory mechanism of homeostasis of their gut microbial communities.

Key words:  Propsilocerus akamusi; gut bacteria, pollution, diversity, function prediction