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徐天梅1, 符成悦1, 苏造堂1, 肖关丽2*, 李维薇1,3, 陈 斌1*.云南草地贪夜蛾扩散区与首发地种群肠道细菌群落组成结构及多样性的比较[J].植物保护,2020,46(4):116-125.
云南草地贪夜蛾扩散区与首发地种群肠道细菌群落组成结构及多样性的比较
Composition and diversity of gut bacteria community of Spodoptera frugiperda from the first invasion site and the dispersal area in Yunnan province
投稿时间:2020-02-20  修订日期:2020-02-29
DOI:10.16688/j.zwbh.2020083
中文关键词:  草地贪夜蛾  肠道细菌  16S rDNA  基因测序
英文关键词:Spodoptera frugiperda  gut bacteria  16S rDNA  gene sequencing
基金项目:国家重点研发计划(2019YFD0300105,2018YFD0200703);国家自然科学基金(31760519,31660537)
作者单位E-mail
徐天梅1, 符成悦1, 苏造堂1, 肖关丽2*, 李维薇1,3, 陈 斌1* 1. 云南农业大学植物保护学院, 云南生物资源保护与利用国家重点实验室, 昆明 650201
2. 云南农业大学农学与生物技术学院, 昆明 650201
3. 中国科学院昆明动物研究所, 昆明 650223 
肖关丽glxiao9@163.com;陈斌chbins@163.com 
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中文摘要:
      草地贪夜蛾是一种新入侵我国的重大害虫。肠道细菌是昆虫肠道微生物的重要类群,在害虫生理生态功能中发挥着重要作用,为弄清云南省草地贪夜蛾扩散地与入侵地种群肠道细菌群落组成结构及多样性的差异,本研究采用Illumina Hiseq技术对云南草地贪夜蛾首发地普洱市宝藏镇、扩散区昆明市嵩明县两个地区草地贪夜蛾4龄幼虫肠道细菌的16S rDNA V3-V4变异区域序列进行了测定分析,应用USEARCH和QIIME软件统计样品序列数和操作分类单元(operational taxonomic unit,OTU)数量分析了肠道细菌群落组成结构及物种丰度与多样性。结果表明,来自普洱市宝藏镇、昆明市嵩明县的6个草地贪夜蛾样品共产生了302 786条有效序列,在97%相似度下可将其聚成564个OTUs,共发现细菌20门、39纲、86目、161科、318属、344种。其中宝藏镇草地贪夜蛾种群的肠道细菌分属于16门、29纲、60目、102科、178属共192种;优势门、纲、目、科、属、种分别为:厚壁菌门(35.09%)、梭菌纲(15.84%)、梭菌目(15.84%)、艾克曼菌科(12.80%)、艾克曼菌属(12.80%)、不可培养的艾克曼菌属细菌(12.80%)。嵩明县草地贪夜蛾肠道细菌共184种,分属于15门、27纲、54目、96科、172属;优势门、纲、目、科、属、种分别为:厚壁菌门(65.87%)、杆菌纲(48.07%)、乳杆菌目(47.32%)、肠球菌科(46.99%)、肠球菌属(46.99%)、NLAE-zl-C485(46.96%)。宝藏镇草地贪夜蛾4龄幼虫肠道内细菌群落Shannon、ACE和Chao1多样性指数分别为3.46、360.25和370.54,分别高于扩散地嵩明县种群的2.26、295.54和305.42;而Simpson指数(0.13)则低于嵩明县种群(0.36)。通过KEGG代谢途径差异分析发现,草地贪夜蛾肠道细菌基因主要参与的代谢途径是整体组织构建和碳水化合物代谢,且宝藏镇草地贪夜蛾肠道细菌基因在大多数的代谢通路中表达量高于嵩明县种群。以上结果说明,云南省草地贪夜蛾首发地种群肠道内细菌群落组成及多样性高于扩散区种群。本研究将为进一步研究不同地区草地贪夜蛾种群生物学差异及肠道微生物对其生态适应性的调控提供理论依据。
英文摘要:
      Fall armyworm, Spodoptera frugiperda, is a serious invaded pest insect of corn, it was firstly found in Baozang town, Yunnan province, China, in 2019. Gut bacteria played important role in the physiological and ecological function. To understand the difference of composition and diversity of gut bacteria communities in S.frugiperda from the first invasion site and the dispersal area, a comparable study on the intestinal bacteria of 4th instar larvae from Baozang and Songming populations of S.frugiperda was conducted using 16S rDNA V3-V4 variation region sequence based on the Illumina Hiseq technique. The sequence numbers of each sample and the numbers of operational taxonomic unit (OTU) were analyzed using the software of USEARCH and QIIME, and the species composition, abundance and diversity indexes of gut bacteria were determined. A total of 302 786 effective sequences were generated from six samples from two regions, which was aggregated into 564 OTUs under 97% similarity. A total of 20 phylum, 39 classes, 86 orders, 161 families, 318 genera and 344 species bacterial were annotated. For Baozang population, a total of 192 species which belong to 178 genus, 102 families, 60 orders, 29 classes and 16 phylum were identified, among which dominant phylum, class, order, family, genera and species were Firmicutes (35.09%), Clostridia (15.84%), Clostridiales (15.84%), Akkermansiaeae (12.80%), Akkermansia (12.80%) and uncultured Akkermansia bacterium (12.80%), respectively. A total of 184 species which belong to 172 genus, 96 families, 54 orders, 27 classes and 15 phylum were identified in Songming population. Of which, dominant phylum, class, order, family, genera and species were Firmicutes (65.87%), Bacilli (48.07%), Lactobacilliales (47.32%), Enterococcaceae (46.99%), Enterococcus (46.99%)and NLAE-zl-c485 (46.96%), respectively. The diversity indexes of Shannon index (3.46), ACE (360.25)and Chao 1 (370.54) in the Baozang population were higher than those in the Songming population, which were 2.26, 295.54 and 305.42, respectively. But the Simpson index in the Baozang population (0.13) was lower than that in the Songming population (0.36). Analysis of the KEGG metabolism pathway showed that the main metabolism pathway of gut bacterial was the whole tissue construction and carbohydrate metabolism, and the relative expression of gut bacterial genes in Baozang population was higher than that in Songming population. These results showed that the composition and diversity of gut bacterial community of S.frugiperda in the first invasion site was higher than those in the dispersal areas in Yunnan province. This study will provide a theoretical basis for further study on the variance of the biology of different populations and adaptation mechanism meditated by gut microorganisms.
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