摘要
[目的]分析天敌昆虫叉角厉蝽[Eocanthecona furcellata(Wolff)]不同发育阶段肠道细菌群落组成及多样性,并预测其功能,为深入了解肠道细菌对叉角厉蝽生长发育和营养获取方式的影响及开发新的害虫生物防治策略提供理论依据.[方法]选取叉角厉蝽1龄、2龄、5龄若虫和7日龄雌、雄成虫,分别提取其肠道细菌基因组DNA.利用单分子实时测序技术对肠道细菌16S rRNA序列全长进行测序,统计操作分类单元(OTU)数量,分析肠道细菌群落组成结构与多样性,并采用PICRUSt预测肠道细菌功能.[结果]从叉角厉蝽15个肠道样品中共得到202401条有效序列,聚类为222个OTUs,其中共有OTUs为5个.共注释到19门32纲62目103科154属188种细菌,优势门、纲、目、科、属、种分别为变形菌门(Proteobacteria,72.22％)、γ-变形菌纲(Gammaproteobacteria,72.21％)、肠杆菌目(Enterobacterales,72.11％)、肠杆菌科(Enterobacteriaceae,49.39％)、西地西菌属(Cedecea,49.24％)和奈氏西地西菌(C.neteri,49.24％).多样性和差异分析结果显示,肠道细菌菌群组成在叉角厉蝽不同发育阶段存在显著差异(P＜0.05,下同).线性判别分析结果显示,1龄、2龄若虫和7日龄雌、雄成虫肠道具有显著优势的菌群,而5龄若虫肠道不存在显著优势菌群(P＞0.05),且不同发育阶段肠道的优势菌群在科、属、种3级分类阶元水平上亦不同.KEGG代谢途径分析发现,叉角厉蝽不同发育阶段肠道菌群的功能相似,主要涉及新陈代谢(74.17％～75.59％)和环境信息处理(10.53％～11.03％),二级代谢功能通路主要为全局和概述图谱、糖类代谢和氨基酸代谢等.[结论]叉角厉蝽不同发育阶段肠道细菌群落组成和多样性存在显著差异,优势菌分布呈动态变化.
Abstract
[Objective]The purpose of the study was to analyze the composition and diversity of gut bacterial commu-nities in different developmental stages of the natural enemy insect Eocanthecona furcellata(Wolff),and to predict their functions,in order to provide a theoretical basis for a deeper understanding of the influence of gut bacteria on the growth,development and nutrient acquisition patterns of E.furcellata,as well as for the development of a novel biocontrol strate-gies for the pests.[Method]The gut bacterial genomic DNA was extracted from the first instar nymphs,second instar nymphs,fifth instar nymphs,7-day-old female and male adults of E.furcellata,respectively.The full-length of 16S rRNA sequence of E.furcellata gut bacteria was sequenced using the single-molecule real-time sequencing technique.The number of operational taxonomic unit(OTU)was counted,the composition and diversity of gut bacterial communities were analyzed,and the functions of gut bacteria were predicted by PICRUSt.[Result]A total of 202401 effective se-quences were obtained from 15 gut samples of E.furcellata,and clustered into 222 OTUs,of which 5 OTUs were shared by them.A total of 19 phyla,32 classes,62 orders,103 families,154 genera and 188 species bacteria were annotated.The dominant phylum,class,order,family,genus and species of the bacteria were Proteobacteria(72.22％),Gammaproteo-bacteria(72.21％),Enterobacterales(72.11％),Enterobacteriaceae(49.39％),Cedecea(49.24％),and C.neteri(49.24％),respectively.The results of the diversity and variance analysis revealed significant differences in the composition of the gut bacterial flora in different developmental stages of E.furcellata(P＜0.05,the same below).The linear discriminant analysis results showed that significantly dominant flora existed in the guts of first instar nymphs,second instar nymphs and 7-day-old female and male adults,while no significantly dominant flora existed in the gut of fifth instar nymphs(P＞0.05),and the dominant flora in the gut of different developmental stages varied at the level of 3 taxonomic catego-ries,namely,family,genus and species.KEGG metabolic pathway analysis revealed that the functions of the gut flora of E.furcellata were similar in different developmental stages,mainly involving metabolism(74.17％-75.59％)and environ-mental information processing(10.53％-11.03％).The secondary metabolic pathways were mainly global and overview mapping,sugar metabolism,and amino acid metabolism,etc.[Conclusion]Significant differences in the composition and diversity of gut bacterial community are observed in different developmental stages of E.furcellata,and the distribu-tion of dominant bacteria shows dynamic changes.
基金项目
云南省农业基础研究联合专项(202101BD070001-050)
云南省农业基础研究联合专项(202301BD070001-137)
云南省中青年学术和技术带头人后备人才项目(202205AC 160077)
云南省烟草公司科技计划项目(2023530000241012)
维普
万方数据