稻纵卷叶螟肠道细菌群落结构与多样性分析  

Analysis of the bacterial community structure and diversity in the intestine of Cnaphalocrocis medinalis( Lepidoptera: Pyralidae)

作　　者：刘小改[1,2] 杨亚军[2] 廖秋菊[1] 徐红星[2] 刘映红[1] 吕仲贤[2] 

机构地区：[1]西南大学植物保护学院,重庆400716 [2]浙江省农业科学院植物保护与微生物研究所,浙江省植物有害生物防控重点实验室省部共建国家重点实验室培育基地,杭州310021

出　　处：《昆虫学报》2016年第9期965-976,共12页Acta Entomologica Sinica

基　　金：国家水稻产业技术体系(CARS-01-17);国家自然科学基金项目(31501669);浙江省重点研发项目(2015C02014)

摘　　要：【目的】为明确水稻主要害虫稻纵卷叶螟Cnaphalocrocis medinalis（Guenée）幼虫肠道细菌的群落结构和多样性。【方法】利用Illumina Mi Seq技术对稻纵卷叶螟4龄幼虫肠道细菌的16S r DNA V3-V4变异区序列进行测序,应用USEARCH和QIIME软件整理和统计样品序列数和操作分类单元（operational taxonomic unit,OTU）数量,分析4龄幼虫肠道细菌的物种组成、丰度和多样性。【结果】稻纵卷叶螟4个数量不同的4龄幼虫样本（1,2,3和5头）共得165 386条reads,在97%相似度下可将其聚类为604个OTUs。总共注释到22个门,43个纲,82个目,142个科,204个属,244个种。其中在门水平上,主要优势菌为变形菌门（Proteobacteria）（相对丰度26%~34%）和放线菌门（Actinobacteria）（23%~32%）;在纲水平上,主要优势菌为放线菌纲（Actinobacteria）（相对丰度23%~32%）、酸杆菌纲（Acidobacteria）（9%~11%）、α-变形菌纲（Alphaproteobacteria）（10%~13%）、β-变形菌纲（Betaproteobacteria）（6%~8%）和γ-变形菌纲（Gammaproteobacteria）（6%~12%）;在科水平上,共有优势菌为类诺卡氏菌科（Nocardioidaceae）、丛毛单胞菌科（Comamonadaceae）、黄单胞菌科（Xanthomonadaceae）、鞘脂单胞菌科（Sphingomonadaceae）和芽单胞菌科（Gemmatimonadaceae）等。在属水平上,4个样本前5位优势属中,类诺卡氏属Nocardioides和鞘脂单胞菌Sphingomonas为共有优势属。稻纵卷叶螟肠道细菌Simpson指数、Shannon指数、Ace指数和Chao指数分别为0.16~0.65,0.94~3.22,212~488和210~490。【结论】稻纵卷叶螟幼虫肠道细菌多样性比较丰富,个体间微生物群落结构和多样性有差异。不同数量样本数据与总体数据有助于综合反映稻纵卷叶螟种群肠道微生物状况。本研究结果为进一步研究稻纵卷叶螟肠道微生物的功能及其在防治中的应用奠定了基础。【Aim 】 To clarify the community structure and diversity of bacteria in the intestine of Cnaphalocrocis medinalis（ Guenée） larvae. 【Methods】The V3-V4 regions of the 16 S r DNA genes of intestinal bacteria in the 4th instar larvae of C. medinalis were sequenced by Illumina Mi Seq techniques.The numbers of sequences and operational taxonomic units（ OTUs）,species composition,abundance and diversity of the bacteria were analyzed using USEARCH and QIIME softwares. A total of 165 386 reads were obtained in the four samples consisting of 1,2,3 and 5 larvae of C. medinalis,respectively.【Results】After filtration,604 OTUs were acquired and annotated into 22 phyla,43 classes,82 orders,142 families, 204 genera, and 244 species. Common dominant bacteria based on phyla were Proteobacteria（ relative abundance 26%- 34%） and Actinobacteria（ 23%- 32%）. Actinobacteria（ relative abundance 23%- 32%）,Acidobacteria（ 9%- 11%）,Alphaproteobacteria（ 10%- 13%）,Betaproteobacteria（ 6%- 8%）,Gammaproteobacteria（ 6%- 12%） were dominant at the class level.Nocardioidaceae,Comamonadaceae,Xanthomonadaceae,Sphingomonadaceae,and Gemmatimonadaceae were commonly dominated at the family level in the bacterial communities from the four samples,and Nocardioides and Sphingomonas were the commonly dominated genera in the top 5 genera of bacterial communities from the four samples. The indices of Simpson,Shannon,Ace and Chao were 0. 16- 0. 65,0. 94- 3. 22,212- 488 and 210- 490,respectively. 【Conclusion】The results indicate that there are diverse bacteria in the intestine of C. medinalis larvae,and there are differences in bacterial structure and diversity among intestines of C. medinalis individuals. The information from samples of different size and the total sample can promote the comprehensive understanding of the bacterial community in the intestine of C. medinalis population. The information of bacterial abundance and diversity in the intestine of C. medinalis will facilitate the studies on the new tech

关 键 词：稻纵卷叶螟 肠道细菌 多样性 16S RDNA 基因测序 

分 类 号：Q965.8[生物学—昆虫学]