|
|
|
Wing patterning genes of Nilaparvata lugens identification by transcriptome analysis, and their differential expression profile in wing pads between brachypterous and macropterous morphs |
LI Kai-yin, HU Ding-bang, LIU Fang-zhou, LONG Man, LIU Si-yi, ZHAO Jing, HE Yue-ping |
Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, P.R.China |
|
|
摘要 The brown planthopper, Nilaparvata lugens is an economically important pest on rice plants. This species produces macropterous and brachypterous morphs in response to environmental cues, which makes it very difficult to control. The molecular basis of wing patterning in N. lugens is still unknown. It is necessary to identify wing patterning genes of N. lugens, and also to clarify the expression differences of wing patterning genes between macropterous and brachypterous morphs. High-throughput deep sequencing of transcriptome of N. lugens wing pad yielded 116 744 580 raw reads and 113 042 700 clean reads. All the reads were assembled into 55 963 unigenes with an average length of 804 bp. With the E-value cut-off of 1.0E–5,18 359 and 2 883 unigens had hits in NCBI-NR (NCBI non-redundant protein sequences) and NCBI-NT (NCBI nucleotide sequences) databases, respectively. A total of 16 502 unigenes were assigned to GO (gene ontology) classification, 9 709 ungenes were grouped into 26 COG (cluster of orthologous groups of proteins) classifications, and 6 724 unigenes were assigned to different KEGG (Kyoto encyclopedia of genes and genomes) pathways. In total, 56 unigenes which are homologous to wing patterning genes of Drosophila melanogaster or Tribolium castaneum were identified. Out of the 56 unigenes, 24 unigenes were selected, and their expression levels across the five nymphal stages between macropterous strain and brachypterous strain were examined by qRT-PCR. Two-way ANOVA analysis showed that development stage had significant effects on the expression level of all the 24 genes (P<0.05). The expression levels of 8 genes (Nlen, Nlhh, Nlsal, NlAbd-A, Nlwg, Nlvg, Nlexd and NlUbx) were significantly affected by wing morph. This is the first transcriptome analysis of wing pads of hemimetabolous insect, N. lugens. The identified wing patterning genes would be useful resource for future exploration of molecular basis of wing development. The 8 differentially expressed wing patterning genes between macropterous strain and brachypterous strain would contribute to explain molecular mechanism of wing-morph differentiation in N. lugens.
Abstract The brown planthopper, Nilaparvata lugens is an economically important pest on rice plants. This species produces macropterous and brachypterous morphs in response to environmental cues, which makes it very difficult to control. The molecular basis of wing patterning in N. lugens is still unknown. It is necessary to identify wing patterning genes of N. lugens, and also to clarify the expression differences of wing patterning genes between macropterous and brachypterous morphs. High-throughput deep sequencing of transcriptome of N. lugens wing pad yielded 116 744 580 raw reads and 113 042 700 clean reads. All the reads were assembled into 55 963 unigenes with an average length of 804 bp. With the E-value cut-off of 1.0E–5,18 359 and 2 883 unigens had hits in NCBI-NR (NCBI non-redundant protein sequences) and NCBI-NT (NCBI nucleotide sequences) databases, respectively. A total of 16 502 unigenes were assigned to GO (gene ontology) classification, 9 709 ungenes were grouped into 26 COG (cluster of orthologous groups of proteins) classifications, and 6 724 unigenes were assigned to different KEGG (Kyoto encyclopedia of genes and genomes) pathways. In total, 56 unigenes which are homologous to wing patterning genes of Drosophila melanogaster or Tribolium castaneum were identified. Out of the 56 unigenes, 24 unigenes were selected, and their expression levels across the five nymphal stages between macropterous strain and brachypterous strain were examined by qRT-PCR. Two-way ANOVA analysis showed that development stage had significant effects on the expression level of all the 24 genes (P<0.05). The expression levels of 8 genes (Nlen, Nlhh, Nlsal, NlAbd-A, Nlwg, Nlvg, Nlexd and NlUbx) were significantly affected by wing morph. This is the first transcriptome analysis of wing pads of hemimetabolous insect, N. lugens. The identified wing patterning genes would be useful resource for future exploration of molecular basis of wing development. The 8 differentially expressed wing patterning genes between macropterous strain and brachypterous strain would contribute to explain molecular mechanism of wing-morph differentiation in N. lugens.
|
Received: 14 November 2014
Accepted:
|
Fund: This work was supported by the National Natural Science Foundation of China (31171846). |
Corresponding Authors:
HUA Hong-xia,E-mail: huahongxia@mail.hzau.edu.cn
E-mail: huahongxia@mail.hzau.edu.cn
|
About author: LI Kai-yin, E-mail: 970777174@qq.com; HU Ding-bang,E-mail: hudingb@163.com;* These authors contributed equally to this study. |
Cite this article:
LI Kai-yin, HU Ding-bang, LIU Fang-zhou, LONG Man, LIU Si-yi, ZHAO Jing, HE Yue-ping.
2015.
Wing patterning genes of Nilaparvata lugens identification by transcriptome analysis, and their differential expression profile in wing pads between brachypterous and macropterous morphs. Journal of Integrative Agriculture, 14(9): 1796-1807.
|
Abouheif E, Wray G A. 2002. Evolution of the gene networkunderlying wing polyphenism in ants. Science, 297,249-252Aspland S E, White R. 1997. Nucleocytoplasmic localisationof extradenticle protein is spatially regulated throughoutdevelopment in Drosophila. Development, 124, 741-747Azpiazu N, Morata G. 2000. Function and regulation ofhomothorax in the wing imaginal disc of Drosophila.Development, 127, 2685-2693Basler K, Struhl G. 1994. Compartment boundaries and thecontrol of Drosophila limb pattern by Hedgehog protein.Nature, 368, 208-214Bertuso A G, Morooka S, Tojo S. 2002. Sensitive periods forwing development and precocious metamorphosis afterprecocene treatment of the brown planthopper, Nilaparvatalugens. Journal of Insect Physiology, 48, 221-229Brisson J A, Ishikawa A, Miura T. 2010. Wing developmentgenes of the pea aphid and differential gene expressionbetween winged and unwinged morphs. Insect MolecularBiology, 19, 63-73Campbell G, Tomlinson A. 1998. The roles of the homeoboxgenes aristaless and Distal-less in patterning the legs andwings of Drosophila. Development, 125, 4483-4493Campbell G, Tomlinson A. 1999. Transducing the Dppmorphogen gradient in the wing of Drosophila: Regulationof Dpp targets by brinker. Cell, 96, 553-562Campbell G, Weaver T, Tomlinson A. 1993. Axis specification inthe developing Drosophila appendage: The role of wingless,decapentaplegic, and the homeobox gene aristaless. Cell,74, 1113-1123Carroll S B, Weatherbee S D, Langeland J A. 1995. Homeoticgenes and the regulation and evolution of insect wingnumber. Nature, 375, 58-61Casares F, Calleja M, Sánchez-Herrero E. 1996. Functionalsimilarity in appendage specification by the Ultrabithoraxand abdominal-A Drosophila HOX genes. The EMBOJournal, 15, 3934-3942Casares F, Mann R S. 2000. A dual role for homothoraxin inhibiting wing blade development and specifyingproximal wing identities in Drosophila. Development, 127,1499-1508de Celis J F, Llimargas M, Casanova J. 1995. Ventralveinless, the gene encoding the Cf1a transcription factor,links positional information and cell differentiation duringembryonic and imaginal development in Drosophilamelanogaster. Development, 121, 3405-3416Cheng X N, Chen R C, Xi X, Yang L M, Zhu Z L, Wu J C, QianR G, Yang J S. 1979. Studies on the migrations of brownplanthoppers, Nilaparvata lugens Stål. Acta EntomologicaSinica, 22, 1-21 (in Chinese)Cohen B, McGuffin M E, Pfeifle C, Segal D, Cohen S M. 1992.apterous, a gene required for imaginal disc developmentin Drosophila encodes a member of the LIM family ofdevelopmental regulatory proteins. Genes & Development,6, 715-729Couso J P, Bate M, Martínez-Arias A. 1993. A wingless dependent polar coordinate system in Drosophila imaginaldiscs. Science, 259, 484-489Deng H, Zhang J, Li Y, Zheng S, Liu L, Huang L, Xu W H,Palli S R, Feng Q. 2012. Homeodomain POU and Abd-Aproteins regulate the transcription of pupal genes duringmetamorphosis of the silkworm, Bombyx mori. Proceedingsof the National Academy of Sciences of the United Statesof America, 109, 12598-12603Doumpas N, Jékely G, Teleman A A. 2013. Wnt6 is requiredfor maxillary palp formation in Drosophila. BMC Biology,11, 104.Götz S, García-Gómez J M, Terol J, Williams T D, NagarajS H, Nueda M J, Robles M, Talón M, Dopazo J, ConesaA. 2008. High-throughput functional annotation and datamining with the Blast2GO suite. Nucleic Acids Research,36, 3420-3435Grabherr M G, Haas B J, Yassour M, Levin J Z, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q,Chen Z, Mauceli E, Hacohen N, Gnirke A, Rhind N, di PalmaF, Birren B W, Nusbaum C, Lindblad-Toh K, FriedmanN, Regev A. 2011. Full-length transcriptome assemblyfrom RNA-Seq data without a reference genome. NatureBiotechnology, 29, 644-652Grimm S, Pflugfelder G O. 1996. Control of the gene optomotorblindin Drosophila wing development by decapentaplegicand wingless. Science, 271, 1601-1604.Gu S H, Wu K M, Guo Y Y, Pickett J A, Field L M, Zhou J J,Zhang Y J. 2013. Identification of genes expressed in thesex pheromone gland of the black cutworm Agrotis ipsilonwith putative roles in sex pheromone biosynthesis andtransport. BMC Genomics, 14, 636.Halder G, Polaczyk P, Kraus M E, Hudson A, Kim J, LaughonA, Carroll S. 1998. The Vestigial and Scalloped proteins acttogether to directly regulate wing-specific gene expressionin Drosophila. Genes & Development, 12, 3900-3909Iseli C, Jongeneel C V, Bucher P. 1999. ESTScan: A programfor detecting, evaluating, and reconstructing potential codingregions in EST sequences. Proceedings of InternationalConference on Intelligent Systems for Molecular Biology,7, 138-148Johnson R L, Grenier J K, Scott M P. 1995. Patchedoverexpression alters wing disc size and pattern:Transcriptional and post-transcriptional effects on hedgehogtargets. Development, 121, 4161-4170Kim E, Niethammer M, Rothschild A, Jan Y N, Sheng M. 1995.Clustering of Shaker-type K+ channels by interaction with afamily of membrane-associated guanylate kinases. Nature,378, 85-88Kukalová-Peck J. 1983. Origin of the insect wing and wingarticulation from the arthropodan leg. Canadian Journal ofZoology, 61, 1618-1669Livak K G, Schmittgen T D. 2001. Analysis of relative geneexpression data using real-time quantitative PCR and the2−ΔΔCT method. Methods, 25, 402-408Lunde K, Biehs B, Nauber U, Bier E. 1998. The knirps andknirps-related genes organize development of the secondwing vein in Drosophila. Development, 125, 4145-4154Morooka S, Tojo S. 1992. Maintenance and selection of strainsexhibiting specific wing form and body colour under highdensity conditions in the brown planthopper, nilaparvatalugens (Homoptera: Delphacidae). Japanese Society ofApplied Entomology and Zoology, 27, 445-454.Motzny C K, Holmgren R. 1995. The Drosophila cubitusinterruptus protein and its role in the wingless and hedgehogsignal transduction pathways. Mechanisms of Development,52, 137-150Neumann C J, Cohen S M. 1996. A hierarchy of cross-regulationinvolving Notch, wingless, vestigial and cut organizes thedorsal/ventral axis of the Drosophila wing. Development,122, 3477-3485Ng M, Diaz-Benjumea F J, Cohen S M. 1995. Nubbin encodes aPOU-domain protein required for proximal-distal patterningin the Drosophila wing. Development, 121, 589-599Panganiban G., Rubenstein J L R. 2002. Developmentalfunctions of the Distal-less/Dlx homeobox genes.Development, 129, 4371-4386Pavlopoulos A, Akam M. 2011. Hox gene Ultrabithoraxregulates distinct sets of target genes at successive stagesof Drosophila haltere morphogenesis. Proceedings of theNational Academy of Sciences of the United States ofAmerica, 108, 2855-2860Posakony L G, Raftery L A, Gelbart W M. 1990. Wing formationin Drosophila melanogaster requires decapentaplegic genefunction along the anterior-posterior compartment boundary.Mechanisms of Development, 33, 69-82Rogers B T, Peterson M D, Kaufman T C. 1997. Evolution ofthe insect body plan as revealed by the Sex combs reducedexpression pattern. Development, 124, 149-157Skeath J B, Carroll S B. 1991. Regulation of achaete-scutegene expression and sensory organ pattern formation inthe Drosophila wing. Genes & Development, 5, 984-995Snodgrass R E. 1935. Principles of Insect Morphology. McGraw-Hill, New York.Soanes K H, MacKay J O, Core N, Heslip T, Kerridge S, BellJ B. 2001. Identification of a regulatory allele of teashirt(tsh) in Drosophila melanogaster that affects wing hingedevelopment. An adult-specific tsh enhancer in Drosophila.Mechanisms of Development, 105, 145-151Sturtevant M A, Biehs B, Marin E, Bier E. 1997. The spaltgene links the A/P compartment boundary to a linear adultstructure in the Drosophila wing. Development, 124, 21-32Syobu S, Mikuriya H, Yamaguchi J, Matsuzaki M, MatsumuraM. 2002. Fluctuations and factors affecting the wing-formratio of the brown planthopper, Nilaparvata lugens Stål inrice fields. Japanese Journal of Applied Entomology andZoology, 46, 135-143Tabata T, Eaton S, Kornberg T B. 1992. The Drosophilahedgehog gene is expressed specifically in posteriorcompartment cells and is a target of engrailed regulation.Genes & Development, 6, 2635-2645 Tabata T, Kornbert T B. 1994. Hedgehog is a signaling proteinwith a key role in patterning Drosophila imaginal discs.Cell, 76, 89-102Tomoyasu Y, Wheeler S R, Denell R E. 2005. Ultrabithoraxis required for membranous wing identity in the beetleTribolium castaneum. Nature, 433, 643-647Torres A F, Huang C, Chong C M, Leung S W, Prieto-da-SilvaA R, Havt A, Quinet Y P, Martins A M, Lee S M, Rádis-Baptista G. 2014. Transcriptome analysis in venom glandof the predatory giant ant Dinoponera quadriceps: insightsinto the polypeptide toxin arsenal of hymenopterans. PLOSONE, 9, e87556.Tribolium Genome Sequencing Consortium. 2008. The genomeof the model beetle and pest Tribolium castaneum. Nature,452, 949-955Tsuneizumi K, Nakayama T, Kamoshida Y, Kornberg T B,Christian J L, Tabata T. 1997. Daughters against dppmodulates dpp organizing activity in Drosophila wingdevelopment. Nature, 389, 627-631Villa-Cuesta E, González-Pérez E, Modolell J. 2007. Appositionof iroquois expressing and non-expressing cells leads to cellsorting and fold formation in the Drosophila imaginal wingdisc. BMC Developmental Biology, 7, 106.Weatherbee S D, Halder G, Kim J, Hudson A, Carroll S. 1998.Ultrabithorax regulates genes at several levels of thewing-patterning hierarchy to shape the development of theDrosophila haltere. Genes & Development, 12, 1474-1482Wigglesworth V B. 1973. Evolution of insect wings and flight.Nature, 246, 127-129Williams J, Bell J, Carroll S. 1991. Control of Drosophila wingand haltere development by the nuclear vestigial geneproduct. Genes & Development, 5, 2481-2495Wu G R, Yu X P, Tao L Y. 1997. Long-term forecast on theoutbreak of brown planthopper (Nilaparvata lugens Stål)and white-backed planthopper (Sogatella furcifera Horvath).Agricultural Sciences in China, 30, 25-29Wu J, Cohen S M. 1999. Proximodistal axis formation inthe Drosophila leg: Subdivision into proximal and distaldomains by Homothorax and Distal-less. Development,126, 109-117Ye J, Fang L, Zheng H K, Zhang Y, Chen J, Zhang Z J, WangJ, Li S T, Li R Q, Bolund L, Wang J. 2006. WEGO: A webtool for plotting GO annotations. Nucleic Acids Research,34, W293-W297.Zhou S S, Sun Z, Ma W, Chen W, Wang M Q. 2014. Denovo analysis of the Nilaparvata lugens (Stål) antennatranscriptome and expression patterns of olfactory genes.Comparative Biochemistry and Physiology (Part D:Genomics & Proteomics), 9, 31–39. |
No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
|
Shared |
|
|
|
|
|
Discussed |
|
|
|
|