基于16S rDNA序列的小贯小绿叶蝉共生细菌多样性研究
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摘要
利用Illumina HiSeq技术,对赤壁、大悟、武汉、咸安和英山5个茶园小贯小绿叶蝉地理种群的成虫共生细菌16SrDNA-V4变异区进行测序,应用Uparse和RDP Classifier等软件统计和分析样本的物种组成、丰度和多样性。5个地区小贯小绿叶蝉成虫的16SrDNA基因序列文库共获得239 645条有效tags,在97%相似阈值下将其聚类为3 403个OTUs。共注释到41个门,116个纲,197个目,272个科,372个属,105个种。5个样本的共生菌在不同分类水平上的组成有所不同,其中在门水平上,主要优势菌为变形菌门Proteobacteria(相对丰度60.6%~97.1%);在纲水平上,相对丰度排名前5位中共有的优势菌为γ-变形菌纲Gammaproteobacteria和α-变形菌纲Alphaproteobacteria;在属水平上,排名前10位中5个样本共有的优势菌为盐单胞菌属Halomonas、希瓦氏菌属Shewanella和沃尔巴克氏体属Wolbachia。小贯小绿叶蝉成虫细菌的Chao指数、Ace指数、Shannon指数和Simpson指数分别为1 065.55~2 841.89,1 130.76~2 914.90,1.07~8.63和0.18~0.99。茶园小贯小绿叶蝉成虫共生细菌多样性比较丰富,不同地理种群的小贯小绿叶蝉细菌群落结构和多样性有差异。本研究结果为进一步研究细菌对小贯小绿叶蝉种群生物学的影响奠定了基础。
The V4 regions of the 16 S rDNA of symbiotic bacteria in adults of Empoasca(Matsumurasca)onukii from five geographical regions including Chibi,Dawu,Wuhan,Xian'an and Yingshan were sequenced by Illumina HiSeq techniques.The numbers of operational taxonomic units(OTUs),bacterial community,species abundance and alpha diversity in samples were analyzed by using Uparse and RDP Classifier software.The results showed that a total of 239 645 effective tags were obtained,which were clustered into3 403 OTUs,and annotated into41 phyla,116 classes,197 orders,272 families,372 genera,and105 species.At the phyla level,Proteobacteria was the common dominant bacteria(relative abundance 60.6%-97.1%).Gammaproteobacteria and Alphaproteobacteria were the commonly dominant classes in the top 5 classes of bacterial communities between the five samples.Halomonas,Shewanella and Wolbachia were the commonly dominant genera in the top 10 genera of bacterial communities.The indexes of Chao,Ace,Shannon and Simpson were 1 065.55-2 841.89,1 130.76-2 914.90,1.07-8.63 and0.18-0.99,respectively.The results indicated that there are diverse bacteria in adults of E.(M.)onukii,and there are differences in the bacterial community diversity among E.(M.)onukii populations.These results provide a basis for further study of the effects of bacteria on the biology of E.(M.)onukii.
The V4 regions of the 16 S rDNA of symbiotic bacteria in adults of Empoasca(Matsumurasca)onukii from five geographical regions including Chibi,Dawu,Wuhan,Xian'an and Yingshan were sequenced by Illumina HiSeq techniques.The numbers of operational taxonomic units(OTUs),bacterial community,species abundance and alpha diversity in samples were analyzed by using Uparse and RDP Classifier software.The results showed that a total of 239 645 effective tags were obtained,which were clustered into3 403 OTUs,and annotated into41 phyla,116 classes,197 orders,272 families,372 genera,and105 species.At the phyla level,Proteobacteria was the common dominant bacteria(relative abundance 60.6%-97.1%).Gammaproteobacteria and Alphaproteobacteria were the commonly dominant classes in the top 5 classes of bacterial communities between the five samples.Halomonas,Shewanella and Wolbachia were the commonly dominant genera in the top 10 genera of bacterial communities.The indexes of Chao,Ace,Shannon and Simpson were 1 065.55-2 841.89,1 130.76-2 914.90,1.07-8.63 and0.18-0.99,respectively.The results indicated that there are diverse bacteria in adults of E.(M.)onukii,and there are differences in the bacterial community diversity among E.(M.)onukii populations.These results provide a basis for further study of the effects of bacteria on the biology of E.(M.)onukii.
引文
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[4]VORBURGER C.The evolutionary ecology of symbiont-conferred resistance to parasitoids in aphids[J].Insect Science,2014,21(3):251-264.
[5]HIMLER A G,ADACHI-HAGIMORI T,BERGEN J E,et al.Rapid spread of a bacterial symbiont in an invasive whitefly is driven by fitness benefits and female bias[J].Science,2011,332(6026):254-256.
[6]单红伟,刘树生,刘银泉.昆虫共同体对不利温度的响应[J].植物保护学报,2016,43(1):24-31.
[7]TSUCHIDA T,KOGA R,MATSUMOTO S,et al.Interspecific symbiont transfection confers a novel ecological trait to the recipient insect[J].Biology Letters,2011,7(2):245-248.
[8]陈宗懋.茶园病虫区系的构成和演替[J].中国茶叶,1979(1):6-8.
[9]EDGAR R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.
[10]DILLON R J,WEBSTER G,WEIGHTMAN A J,et al.Diversity of gut microbiota increases with aging and starvation in the desert locust[J].Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology,2010,97(1):69-77.
[11]GRENIER A M,NARDON C,RAHBE Y.Observations on the microorganisms occurring in the gut of the pea aphid Acyrthosiphon pisum[J].Entomologia Experimentalis et Applicata,1994,70(1):91-96.
[12]靳亮,王金昌,王洪秀,等.16SrRNA基因的PCR-DGGE技术分析茶尺蠖幼虫肠道细菌种群结构及多样性[J].江西科学,2013,31(6):759-763.
[13]刘小改,杨亚军,廖秋菊,等.稻纵卷叶螟肠道细菌群落结构与多样性分析[J].昆虫学报,2016,59(9):965-976.
[14]SCHLOSS P D,DELALIBERA I,HANDELSMAN J O,et al.Bacteria associated with the guts of two wood-boring beetles:Anoplophora glabripennis and Saperda vestita(Cerambycidae)[J].Environmental Entomology,2006,35(3):625-629.
[15]BEHAR A,YUVAL B,JURKEVITCH E.Gut bacterial communities in the Mediterranean fruit fly(Ceratitis capitata)and their impact on host longevity[J].Journal of Insect Physiology,2008,54(9):1377-1383.
[16]张某,杨璞,朱家颖,等.基于16SrDNA基因序列的泽兰实蝇幼虫肠道细菌多样性分析[J].昆虫学报,2016,59(2):200-208.
[17]王娇莉.核桃举肢蛾Atrijuglans hetaohei和桃蛀螟Dichocrocis punctiferalis幼虫肠道细菌组成及多样性研究[D].杨凌:西北农林科技大学,2016.
[18]GLASER R L,MEOLA M A.The native Wolbachia endosymbionts of Drosophila melanogaster and Culex quinquefasciatus increase host resistance to West Nile virus infection[J].PLoS ONE,2010,5(8):e11977.
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