基于RNA-Seq高通量测序技术的菠萝洁粉蚧转录组研究
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摘要
为了探索调节昆虫世代历期的相关基因,研究了菠萝洁粉蚧在转录水平上对温度的响应。组装后获得49 898个Unigenes,其平均长度为1 007 bp;其中有15 015 (30.10%)个Unigenes被成功注释到Nr、SwissProt、KOG和KO等功能数据库中。DEG分析表明,7 960个Unigenes被诱导差异表达,其中MIX21相对于MIX27表达量上调的有5 080个,下调的有2 880个。差异基因代谢通路分析表明,差异表达基因中涉及长寿调节途径(Longevity regulating pathway)的有3个通路,其中21℃饲养下的菠萝洁粉蚧种群胰岛素/IFG-1信号通路关键因子DAF-2、雷帕霉素靶标TOR信号通路关键因子RSK-1和S6K显著下调,相关因子被抑制表达,很好地解释了其世代发育历期比27℃饲养下长的现象。21℃饲养下的菠萝洁粉蚧种群HSP60较27℃饲养下的显著上调,而HSPs有上调也有下调,显示热休克蛋白HSP对温度的反应呈现一定的多样性、复杂性。这些发现将有助于今后昆虫寿命分子调节机制的研究,DAF-2、S6K、RSK-1等相关基因未来或将应用于调节一些经济昆虫的生长周期。
In order to explore the genes related to generation period of the insect, we studied the transcriptomic response of D. brevipes to temperature. We obtained 49 898 assembled unigenes, with average length of 1 007 bp,15 015(30.10%) of which were annotated in Nr, SwissProt, KOG or KO cDNA libraries. The analysis of differentially expressed genes(DEG) showed that 7 960 unigenes were differentially expressed, of which 5 080 unigenes were up-regulated and 2 880 unigenes were down-regulated significantly compared to MIX27 population. As for KO annotation, there were three corresponding pathways longevity regulating. The key factors for signaling pathways of insulin/insulin-like growth factor 1 and target of rapamycin, DAF-2, RSK-1 and S6 K of MIX21 population were down-regulated significantly, their expression were suppressed, which was probably the right explanation for its longer generation period and longevity compared to MIX27 population. The relevant genes for heat-shock protein were complicated, with up-regulated significantly of HSP60 and up-regulated or down-regulated significantly of HSPs compared to MIX27 population. The founding contributed to explore the molecular regulating mechanism of longevity of insect, these related genes, DAF-2, S6 K and RSK-1 might be applied to regulate the growth period of economic insects in the future.
In order to explore the genes related to generation period of the insect, we studied the transcriptomic response of D. brevipes to temperature. We obtained 49 898 assembled unigenes, with average length of 1 007 bp,15 015(30.10%) of which were annotated in Nr, SwissProt, KOG or KO cDNA libraries. The analysis of differentially expressed genes(DEG) showed that 7 960 unigenes were differentially expressed, of which 5 080 unigenes were up-regulated and 2 880 unigenes were down-regulated significantly compared to MIX27 population. As for KO annotation, there were three corresponding pathways longevity regulating. The key factors for signaling pathways of insulin/insulin-like growth factor 1 and target of rapamycin, DAF-2, RSK-1 and S6 K of MIX21 population were down-regulated significantly, their expression were suppressed, which was probably the right explanation for its longer generation period and longevity compared to MIX27 population. The relevant genes for heat-shock protein were complicated, with up-regulated significantly of HSP60 and up-regulated or down-regulated significantly of HSPs compared to MIX27 population. The founding contributed to explore the molecular regulating mechanism of longevity of insect, these related genes, DAF-2, S6 K and RSK-1 might be applied to regulate the growth period of economic insects in the future.
引文
Anders S.,and Huber W.,2010,Differential expression analysis for sequence count data,Genome Biology,11(10):R106
Beardsley J.W.,1965,Notes on the Pineapple mealybug complex,with descriptions of two new species(Homoptera:Pseudococcidae),Proceddings of the Hawaiian Entomological Society,19(1):55-68
Chen D.,Li P.W.L.,Goldstein Benjamin A.,Cai W.J.,Thomas E.L.,Chen F.,Hubbard A.E.,Melov S.,and Kapahi P.,2013,Germline signaling mediates the synergistically prolonged longevity produced by double mutations in daf-2 and rsks-1in C.elegans,Cell Reports,5:1600-1610
Colen K.G.F.,Santacecília L.C.V.,and Moraes J.C.,2000,Effects of different temperatures upon the biology of the mealybug Dysmicoccus brevipes(Cockerell,1893)(Hemiptera:Pseudococcidae),Revista Brasileira De Fruticultura,22:248-252
Grabherr M.G.,Haas B.J.,and Yassour M.,2011,Full-length transcriptome assembly from RNA-Seq data without a reference genome,Nature Biotechnology,29(7):644-652
Han C.,Xie W.H.,Li J.G.,Qin Y.T.,Jiang M.B.,Liu S.Y.H.,and Wang X.K.,2015,Gene expression profile of haloxylon branch under the stresses of drought and deat in summer season,Jiyinzuxue Yu Yingyong Shengwuxue(Chinese Journal of Genomics and Applied Biology),34(3):607-614(韩超,谢文华,李建贵,秦韵婷,蒋劢博,刘隋赟昊,王宪奎,2015,夏季高温干旱逆境下梭梭枝条中转录组表达谱分析,基因组学与应用生物学,34(3):607-614)
He Y.B.,2012,Molecular identification,genetic structure and control strategy of pink pineapple mealybug,Dysmicoccus brevipes(Cockerell),Dissertation for Ph.D.,Southwest University,导师:刘映红,许再福,pp.69-73(何衍彪,2012,菠萝洁粉蚧的分子鉴定、遗传结构及其控制基础研究,博士学位论文,西南大学,Supervisors:Liu Y.H.,Xu Z.F.,pp.69-73)
He Y.B,Wan X.W,Liu Y.H,Sun G.M,and Zhan R.L.,2012,Mitochondrial COI from Dysmicoccus brevipes(Hemiptera:Pseudococcidae)suggests cryptic lineage and pinpoints the source of the introduction to China,Florida Entomologist,95(1):183-191
Ito K.,1938,Studies on the life histories of the pineapple mealybug,Pseudococcus brevipes(Ckll.),Journal of Economic Entomology,31(2):291-298
Pandey R.R.,and Johnson M.W.,2006,Enhanced production of pink pineapple mealybug,Dysmicoccus brevipes(Hemiptera:Pseudococcidae),Biocontrol Science and Technology,16(4):389-401
Qin Z.Q.,Qiu B.L,Wu J.H.,Ags C.,and Ren S.X.,2013,Effect of temperature on the life history of Dysmicoccus neobrevipes(Hemiptera:Pseudoccocidae):an invasive species of gray pineapple mealybug in South China,Crop Protection,45:141-146
Santa-Cecília L.V.C.,Bueno V.H.P.,and Prado E.,2004,Desenvolvimento de Dysmicoccus brevipes(Cockerell)(Hemptera:Pseudococcidae)em duas cultivares de abacaxi,Ciência EAgrotecnologia,28(5):1015-1020
Sarkar S.,and Lakhotia S.C.,2005,The Hsp60C gene in the 25Fcytogenetic region in drosophila melanogaster is essential for tracheal development and fertility,Journal of Genetics,84:265-281
Sether D.M.,and Hu J.S.,2002,Closterovirus infection and mealybug exposure are necessary for the development of mealybug wilt of pineapple disease,Phytopathology 92(9):928-935
Sether D.M.,Melzer M.J.,Busto J.,Zee F.,and Hu J.S.,2005,Diversity and mealybug transmissibility of ampeloviruses in pineapple,Plant Disease,89(5):450-456
Wang H.H.,Reitz S.R.,Wang L.H.,Wang S.Y.,Li X.,Lei Z.R.,2014,The m RNA expression profiles of five heat shock protein genes from Frankliniella occidentalis at different stages and their responses to temperatures and insecticides,Journal of Integrative Agriculture,13(10):2196-2210
Wei C.P.,Jiang J.,Li X.R.,Zhang Y.H.,Zhang F.M.,Cheng D.F.,Guo W.C.,and Liu H.,2017,Cloning and sequence analysis of heat shock protein gene Ld-HSP60 in Leptinotarsa decemlineata(Coleoptera:Chrysomelidae)and its expression under temperature stress,Kunchong Xuebao(Chinese Journal of Acta Entomologica Sinica),60(5):523-532(魏长平,蒋健,李祥瑞,张云慧,张方梅,程登发,郭文超,刘怀,2017,马铃薯甲虫热激蛋白基因Ld-HSP60的克隆,序列分析及温度胁迫下的表达,昆虫学报,60(5):523-532)
Beardsley J.W.,1965,Notes on the Pineapple mealybug complex,with descriptions of two new species(Homoptera:Pseudococcidae),Proceddings of the Hawaiian Entomological Society,19(1):55-68
Chen D.,Li P.W.L.,Goldstein Benjamin A.,Cai W.J.,Thomas E.L.,Chen F.,Hubbard A.E.,Melov S.,and Kapahi P.,2013,Germline signaling mediates the synergistically prolonged longevity produced by double mutations in daf-2 and rsks-1in C.elegans,Cell Reports,5:1600-1610
Colen K.G.F.,Santacecília L.C.V.,and Moraes J.C.,2000,Effects of different temperatures upon the biology of the mealybug Dysmicoccus brevipes(Cockerell,1893)(Hemiptera:Pseudococcidae),Revista Brasileira De Fruticultura,22:248-252
Grabherr M.G.,Haas B.J.,and Yassour M.,2011,Full-length transcriptome assembly from RNA-Seq data without a reference genome,Nature Biotechnology,29(7):644-652
Han C.,Xie W.H.,Li J.G.,Qin Y.T.,Jiang M.B.,Liu S.Y.H.,and Wang X.K.,2015,Gene expression profile of haloxylon branch under the stresses of drought and deat in summer season,Jiyinzuxue Yu Yingyong Shengwuxue(Chinese Journal of Genomics and Applied Biology),34(3):607-614(韩超,谢文华,李建贵,秦韵婷,蒋劢博,刘隋赟昊,王宪奎,2015,夏季高温干旱逆境下梭梭枝条中转录组表达谱分析,基因组学与应用生物学,34(3):607-614)
He Y.B.,2012,Molecular identification,genetic structure and control strategy of pink pineapple mealybug,Dysmicoccus brevipes(Cockerell),Dissertation for Ph.D.,Southwest University,导师:刘映红,许再福,pp.69-73(何衍彪,2012,菠萝洁粉蚧的分子鉴定、遗传结构及其控制基础研究,博士学位论文,西南大学,Supervisors:Liu Y.H.,Xu Z.F.,pp.69-73)
He Y.B,Wan X.W,Liu Y.H,Sun G.M,and Zhan R.L.,2012,Mitochondrial COI from Dysmicoccus brevipes(Hemiptera:Pseudococcidae)suggests cryptic lineage and pinpoints the source of the introduction to China,Florida Entomologist,95(1):183-191
Ito K.,1938,Studies on the life histories of the pineapple mealybug,Pseudococcus brevipes(Ckll.),Journal of Economic Entomology,31(2):291-298
Pandey R.R.,and Johnson M.W.,2006,Enhanced production of pink pineapple mealybug,Dysmicoccus brevipes(Hemiptera:Pseudococcidae),Biocontrol Science and Technology,16(4):389-401
Qin Z.Q.,Qiu B.L,Wu J.H.,Ags C.,and Ren S.X.,2013,Effect of temperature on the life history of Dysmicoccus neobrevipes(Hemiptera:Pseudoccocidae):an invasive species of gray pineapple mealybug in South China,Crop Protection,45:141-146
Santa-Cecília L.V.C.,Bueno V.H.P.,and Prado E.,2004,Desenvolvimento de Dysmicoccus brevipes(Cockerell)(Hemptera:Pseudococcidae)em duas cultivares de abacaxi,Ciência EAgrotecnologia,28(5):1015-1020
Sarkar S.,and Lakhotia S.C.,2005,The Hsp60C gene in the 25Fcytogenetic region in drosophila melanogaster is essential for tracheal development and fertility,Journal of Genetics,84:265-281
Sether D.M.,and Hu J.S.,2002,Closterovirus infection and mealybug exposure are necessary for the development of mealybug wilt of pineapple disease,Phytopathology 92(9):928-935
Sether D.M.,Melzer M.J.,Busto J.,Zee F.,and Hu J.S.,2005,Diversity and mealybug transmissibility of ampeloviruses in pineapple,Plant Disease,89(5):450-456
Wang H.H.,Reitz S.R.,Wang L.H.,Wang S.Y.,Li X.,Lei Z.R.,2014,The m RNA expression profiles of five heat shock protein genes from Frankliniella occidentalis at different stages and their responses to temperatures and insecticides,Journal of Integrative Agriculture,13(10):2196-2210
Wei C.P.,Jiang J.,Li X.R.,Zhang Y.H.,Zhang F.M.,Cheng D.F.,Guo W.C.,and Liu H.,2017,Cloning and sequence analysis of heat shock protein gene Ld-HSP60 in Leptinotarsa decemlineata(Coleoptera:Chrysomelidae)and its expression under temperature stress,Kunchong Xuebao(Chinese Journal of Acta Entomologica Sinica),60(5):523-532(魏长平,蒋健,李祥瑞,张云慧,张方梅,程登发,郭文超,刘怀,2017,马铃薯甲虫热激蛋白基因Ld-HSP60的克隆,序列分析及温度胁迫下的表达,昆虫学报,60(5):523-532)