上海市松江区白纹伊蚊共生微生物宏基因组分析

doi:10.20199/j.issn.1672-2302.2024.06.003

摘要/Abstract

摘要：

目的研究上海市松江区野外环境中白纹伊蚊共生微生物群物种组成和功能的多样性，了解白纹伊蚊传播疾病的现况，为白纹伊蚊生物防制提供新思路。方法 2023年7—9月，在上海市松江区广富林郊野公园、佘山国家森林公园、辰山等3个野外环境采样点使用人诱停落法采集白纹伊蚊样本，形态学鉴定后提取DNA，使用宏基因组测序技术进行样本物种注释，通过综合抗生素耐药性数据库（Comprehensive Antibiotic Resistance Database, CARD）、碳水化合物活性酶数据库（Carbohydrate-Active enZymes Database, CAZy）进行功能注释。结果共捕获白纹伊蚊417只，注释到236个细菌属和23个病毒属，其中3处采样点共有的细菌属占52.97%（125/236），共有的病毒微生物属占21.73%（5/23）。白纹伊蚊样本中优势细菌门为假单胞菌门（Pseudomonadota）和芽孢杆菌门（Bacillota），相对丰度较高的细菌属为沃尔巴克氏体属（Wolbachia）、链球菌属（Streptococcus）和肠杆菌属（Enterobacter）。除去无法分类的微生物病毒门，样本中最主要的优势病毒门为反链RNA病毒门（Negarnaviricota）和衣壳病毒门（Peploviricota），相对丰度较高的病毒属为α裸病毒属（Alphanudivirus）和正黏黄病毒属（Orthoflavivirus）。经CARD数据库对抗生素抗性基因进行注释，结果显示fabG、abcA、rpoB等基因丰度较高;经CAZy数据库对碳水化合物活性酶功能进行注释，结果显示糖苷转移酶类（glycosyl transferases, GT）、糖苷水解酶类（glycoside hydrolases, GH）和多糖裂解酶类（polysaccharide lyases, PL）丰度较高。结论上海市松江区白纹伊蚊共生微生物群构成相似度高，核心菌群稳定，且白纹伊蚊共生微生物群与白纹伊蚊抗药性的形成可能存在潜在关联。

关键词: 白纹伊蚊, 共生微生物群, 宏基因组学, 数据库, 功能注释

Abstract:

ObjectiveTo preliminarily investigate the species diversity and functional variations of the symbiotic microflora in Aedes albopictus within the field environment of Songjiang District, Shanghai Municipality for understanding the changes of disease spectrum transmitted by Ae. albopictus and providing new ideas for biological control of Ae. albopictus. Methods Between July and September 2023, we harvested the Ae. albopictus samples by human trap method from the three field sampling sites in Guangfulin Country Park, Sheshan National Forest Park and Chenshan area, Songjiang District, Shanghai. After morphological identification of the specimens, DNA was extracted, and metagenomic sequencing technology was used to annotate the sample species. Functional annotation was performed through the Comprehensive Antibiotic Resistance Database (CARD) and Carbohydrate-Active enZymes Database (CAZy). Results In this investigation, we successfully captured 417 heads of Ae. albopictus in total, and annotated 236 bacterial genera and 23 viral genera, of which 52.97% (125/236) of the bacterial genera and 21.73% (5/23) of the viral genera were shared in the Ae. albopictus from 3 sampling sites. The dominant phyla of the symbiotic microbiota were Pseudomonadota and Bacillota, with Wolbachia, Streptococcus and Enterobacter showing the higher abundance. Exclusion of the unclassifiable microbial virus phylum, the prevailing virus phyla in the sampls consisted of Negarnaviricota and Peploviricota. Genera with relatively higher abundance included Alphanudivirus and Orthoflavivirus. Annotating the antibiotic resistance genes via CARD database showed that the abundance of fabG, abcA, rpoB and other pathways were higher. Annotation of the function of carbohydrate active enzymes based on CAZy database revealed higher abundance of glycosyl transferases (GT), glycoside hydrolases (GH) and polysaccharide lyases (PL). Conclusion The composition of symbiotic microbiota in Ae. albopictus in Songjiang District of Shanghai is highly similar and the core symbiotic microbiota remains stable. The symbiotic microbiota of Ae. albopictus is potentially associated with the drug resistance.

Key words: Aedes albopictus, Symbiotic microbiota, Metagenomics, Database, Function annotation

中图分类号:

R373.3

叶独秋, 张驰, 庞博文, 吴佳玲, 蒋露芳, 吕锡宏. 上海市松江区白纹伊蚊共生微生物宏基因组分析[J]. 热带病与寄生虫学, 2024, 22(6): 334-338.

YE Duqiu, ZHANG Chi, PANG Bowen, WU Jialing, JIANG Lufang, LÜ Xihong. Metagenomic analysis of symbiotic microbiota in Aedes albopictus from Songjiang District, Shanghai Municipality[J]. Journal of Tropical Diseases and Parasitology, 2024, 22(6): 334-338.

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采样点	采样点经纬度	采样时间
广富林	（31.069 203N，121.183 424E）	2023年7—9月
佘山	（31.094 166N，121.196 041E）	2023年7—8月
辰山	（31.075 000N，121.183 040E）	2023年8—9月

采样点	样本数（只）	原始读段数	清洗后读段数	读段长度（nt）	Q20（%）	GC含量（%）
广富林	136	92 876 170	91 253 886	150.00	97.70	40.76
佘山	143	100 300 226	91 312 476	150.00	97.96	40.80
辰山	138	85 520 800	85 296 178	150.00	97.53	40.81
合计	417	278 697 196	267 862 540	150.00	97.73	40.79