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Issue: Vol. 4591 No. 1: 2 May 2019
Type: Monograph
Published: 2019-05-02
DOI: 10.11646/zootaxa.4591.1.1
Page range: 1–71
Abstract views: 84
PDF downloaded: 2

The RNA-seq approach to discriminate gene expression profiles in response to Beauveria bassiana and Micrococcus luteus microbial pathogens on Actias selene (Lepidoptera: Saturniidae)

College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
College of Life Sciences, Anhui Agricultural University, 130 ChangJiang West Road, Hefei, 230036, PR China
Actias selene comparative transcriptology DEGs immune-related genes Lepidoptera

Abstract

Actias selene (Hübner) is an important silk-spinning moth. Like other moths, it has innate immunity but no acquired immunity. However, there are few studies on immune-related genes of A. selene. Here, differential expression RNAseq experiment was employed to examine the genes related to different metabolic pathways and to explore the immune mechanism of the A. selene post Beauveria bassiana (Bb) and Micrococcus luteus (ML) stimuli. A total of 64,372,921 clean reads were obtained and 39,057 differentially expressed genes (DEGs) were identified. In the Bb vs. PBS group (PBS as the control), 9,092 genes were up-regulated and 4,438 genes were down-regulated; in the ML vs. PBS group, 5,903 genes were up-regulated and 5,175 genes were down-regulated. The KEGG (Kyoto Encyclopedia of Genes and Genomes) and GO (Gene Ontology) analyses of DEGs confirmed that many DEGs were associated with "Metabolism pathway", "cellular process", "cell" and "catalytic activity". Among them, 194 and 149 differentially expressed genes were related to immunity in the Bb vs. PBS group and ML vs. PBS group, respectively. We verified the reliability of the data with reverse transcription quantitative real-time PCR analysis of randomly selected genes. Furthermore, the phylogenetic tree results showed that HSP90, PGRP and MyD88 genes of A. selene were most closely related to Antheraea pernyi (Guérin-Méneville). These results will provide an overview of the molecular mechanism of A. selene resistance to fungal and bacterial infections as well as an evolutionary aspect of these genes. Moreover, the interrelated trophic mechanisms among different groups of organisms are vital to explore, thus this study will lay a foundation for further studies on the innate immune mechanism of saturniid moths, and provide important theoretical basis for studying the relationship between A. selene and other species.

References

  1. Ansorge, W.J. (2009) Next-generation DNA sequencing techniques. Nature Biotechnology, 25 (4), 195–203.
    https://doi.org/10.1016/j.nbt.2008.12.009

    Asea, A., Rehli, M., Kabingu, E., Boch, J.A., Bare, O., Auron, P.E., Stevenson, M.A. & Calderwood, S.K. (2002) Novel signal transduction pathway utilized by extracellular HSP70: role of toll-like receptor (TLR) 2 and TLR4. Journal of Biological Chemistry, 277 (17), 15028–15034.
    https://doi.org/10.1074/jbc.M200497200

    Basu, S. & Srivastava, P.K. (2000) Heat shock proteins: the fountainhead of innate and adaptive immune responses. Cell Stress Chaperones, 5 (5), 443–451.

    https://doi.org/10.1379/1466-1268(2000)005<0443:HSPTFO>2.0.CO;2

    Boman, H.G. & Hultmark, D. (1987) Cell-free immunity in insects. Annual Review Microbiology, 41, 103–126.
    http://doi.org/10.1146/annurev.mi.41.100187.000535

    Camon, E., Magrane, M., Barrell, D., Lee, V., Dimmer, E., Maslen, J., Binns, D., Harte, N., Lopez, R. & Apweiler, R. (2004) The Gene Ontology Annotation (GOA) Database: sharing knowledge in Uniprot with Gene Ontology. Nucleic Acids Research, 32 (Database Issue), D262–266.
    https://doi.org/10.1093/nar/gkh021

    Chen, M.M., Li, Y., Chen, M., Wang, H., Li, Q., Xia, R.X., Zeng, C.Y., Li, Y.P., Liu, Y.Q. & Qin, L. (2014) Complete mitochondrial genome of the atlas moth, Attacus atlas (Lepidoptera: Saturniidae) and the phylogenetic relationship of Saturniidae species. Gene, 545 (1), 95–101.
    https://doi.org/10.1016/j.gene.2014.05.002

    Chen, X., Hu, Y., Zheng, H., Cao, L., Niu, D., Yu, D., Sun, Y., Hu, S. & Hu, F. (2012) Transcriptome comparison between honey bee queen- and worker-destined larvae. Insect Biochemistry & Molecular Biology, 42 (9), 665–673.
    https://doi.org/10.1016/j.ibmb.2012.05.004

    Chen, Y.-R., Jiang, T., Zhu, J., Xie, Y.-C., Tan, Z.-C., Chen, Y.-H., Tang, S.-M., Hao, B.-F., Wang, S.-P., Huang, J.-S. & Shen, X.-J. (2017) Transcriptome sequencing reveals potential mechanisms of diapause preparation in bivoltine silkworm Bombyx mori (Lepidoptera: Bombycidae). Comparative Biochemistry and Physiology Part D: Genomics and Proteomics, 24, 68–78.
    https://doi.org/10.1016/j.cbd.2017.07.003

    Chetia, H., Kabiraj, D., Singh, D., Mosahari, P.V., Das, S., Sharma, P., Neog, K., Sharma, S., Jayaprakash, P. & Bora, U. (2017) De novo transcriptome of the muga silkworm, Antheraea assamensis (Helfer). Gene, 611 54–65.
    https://doi.org/10.1016/j.gene.2017.02.021

    Conesa, A., Gotz, S., Garcia-Gomez, J.M., Terol, J., Talon, M. & Robles, M. (2005) Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics, 21 (18), 3674–3676.
    https://doi.org/10.1093/bioinformatics/bti610

    Dai, L.-S., Qian, C., Wang, L., Wei, G.-Q., Liu, Q.-N., Sun, Y., Zhang, C.-F., Li, J., Liu, D.-R., Zhu, B.-J. & Liu, C.-L. (2015) Characterization and function of a short peptidoglycan recognition protein from the Chinese oak silkworm, Antheraea pernyi. Journal of Asia-Pacific Entomology, 18 (4), 701–707.
    https://doi.org/10.1016/j.aspen.2015.08.003

    Fang, S.L., Wang, L., Fang, Q., Chen, C., Zhao, X.S., Qian, C., Wei, G.Q., Zhu, B.J. & Liu, C.L. (2017) Characterization and functional study of a Cecropin-like peptide from the Chinese oak silkworm, Antheraea pernyi. Archives of Insect Biochemistry & Physiology, 94 (1), e21368.
    https://doi.org/10.1002/arch.21368

    Ferre, R., Melo, M.N., Correia, A.D., Feliu, L., Bardaji, E., Planas, M. & Castanho, M. (2009) Synergistic effects of the membrane actions of cecropin-melittin antimicrobial hybrid peptide BP100. Biophysical Journal, 96 (5), 1815–1827.
    https://doi.org/10.1016/j.bpj.2008.11.053

    Gazara, R.K., Cardoso, C., Bellieny-Rabelo, D., Ferreira, C., Terra, W.R. & Venancio, T.M. (2017) De novo transcriptome sequencing and comparative analysis of midgut tissues of four non-model insects pertaining to Hemiptera, Coleoptera, Diptera and Lepidoptera. Gene, 627, 85–93.
    https://doi.org/10.1016/j.gene.2017.06.008

    Gillespie, J.P., Kanost, M.R. & Trenczek, T. (1997) Biological mediators of insect immunity. Annual Review of Entomology, 42, 611–643.
    https://doi.org/10.1146/annurev.ento.42.1.611

    Grabherr, M.G., Haas, B.J., Yassour, M., Levin, J.Z., Thompson, D.A., Amit, I., Adiconis, X., Fan, L., Raychowdhury, R., Zeng, Q., Chen, Z., Mauceli, E., Hacohen, N., Gnirke, A., Rhind, N., di Palma, F., Birren, B.W., Nusbaum, C., Lindblad-Toh, K., Friedman, N. & Regev, A. (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nature Biotechnology, 29, 644.
    https://doi.org/10.1038/nbt.1883

    Guo, X.-j. & Feng, J.-n. (2018) Comparisons of Expression Levels of Heat Shock Proteins (hsp70 and hsp90) From Anaphothrips obscurus (Thysanoptera: Thripidae) in Polymorphic Adults Exposed to Different Heat Shock Treatments. Journal of Insect Science, 18 (3), 15–15.
    https://doi.org/10.1093/jisesa/iey059

    Huang, S., Yang, H., Chen, X., Zhang, J. & Huang, L. (2018) Genomic transcriptional response to 20-hydroxyecdysone in the fat body of silkworm, Bombyx mori. Gene Reports, 13, 134–140.
    https://doi.org/10.1016/j.genrep.2018.09.009

    Iordanskiy, S., Zhao, Y., Dubrovsky, L., Iordanskaya, T., Chen, M., Liang, D. & Bukrinsky, M. (2004) Heat shock protein 70 protects cells from cell cycle arrest and apoptosis induced by human immunodeficiency virus type 1 viral protein R. Journal of Virology, 78 (18), 9697–9704.
    https://doi.org/10.1128/jvi.78.18.9697-9704.2004

    Julian, L. & Olson, M.F. (2014) Rho-associated coiled-coil containing kinases (ROCK): structure, regulation, and functions. Small GTPases, 5 e29846.
    https://doi.org/10.4161/sgtp.29846

    Kaneko, T., Goldman, W.E., Mellroth, P., Steiner, H., Fukase, K., Kusumoto, S., Harley, W., Fox, A., Golenbock, D. & Silverman, N. (2004) Monomeric and polymeric gram-negative peptidoglycan but not purified LPS stimulate the Drosophila IMD pathway. Immunity, 20 (5), 637–649.
    https://doi.org/10.1016/S1074-7613(04)00104-9

    Kim, J.G., Moon, M.Y., Kim, H.J., Li, Y., Song, D.K., Kim, J.S., Lee, J.Y., Kim, J., Kim, S.C. & Park, J.B. (2012) Ras-related GTPases Rap1 and RhoA collectively induce the phagocytosis of serum-opsonized zymosan particles in macrophages. Journal of Biological Chemistry, 287 (7), 5145–5155.
    https://doi.org/10.1074/jbc.M111.257634

    Kitching, I. & Rawlins, J. (1998) Handbook of Zoology. In: Kristensen, N.P. (Ed.), Lepidoptera, Moths and Butterflies. Vol. 1. Evolution, Systematics and Biogeography. Walter de Gruyter, Berlin, pp. 355–401.

    Liu, Q.-N., Zhu, B.-J., Dai, L.-S., Wei, G.-Q. & Liu, C.-L. (2012) The complete mitochondrial genome of the wild silkworm moth, Actias selene. Gene, 505 (2), 291–299.
    https://doi.org/10.1016/j.gene.2012.06.003

    Livak, K.J. & Schmittgen, T.D. (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25 (4), 402–408.
    https://doi.org/10.1006/meth.2001.1262

    Lu, D., Geng, T., Hou, C., Huang, Y., Qin, G. & Guo, X. (2016) Bombyx mori cecropin A has a high antifungal activity to entomopathogenic fungus Beauveria bassiana. Gene, 583 (1), 29–35.
    https://doi.org/10.1016/j.gene.2016.02.045

    Luan, J.B., Li, J.M., Varela, N., Wang, Y.L., Li, F.F., Bao, Y.Y., Zhang, C.X., Liu, S.S. & Wang, X.W. (2011) Global analysis of the transcriptional response of whitefly to tomato yellow leaf curl China virus reveals the relationship of coevolved adaptations. Journal of Virology, 85 (7), 3330–3340.
    https://doi.org/10.1128/jvi.02507-10

    Ma, J., Xue, Y., Cui, W., Li, Y., Zhao, Q., Ye, W., Zheng, J., Cheng, Y., Ma, Y., Li, S., Han, T., Miao, L., Yao, L., Zhang, J. & Liu, W. (2012) Ras homolog gene family, member A promotes p53 degradation and vascular endothelial growth factor-dependent angiogenesis through an interaction with murine double minute 2 under hypoxic conditions. Cancer, 118 (17), 4105–4116.
    https://doi.org/10.1002/cncr.27393

    Mason, F.M., Xie, S., Vasquez, C.G., Tworoger, M. & Martin, A.C. (2016) RhoA GTPase inhibition organizes contraction during epithelial morphogenesis. Journal of Cell Biology, 214 (5), 603–617.
    https://doi.org/10.1083/jcb.201603077

    Mortazavi, A., Williams, B.A., McCue, K., Schaeffer, L. & Wold, B. (2008) Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nature Methods, 5 (7), 621–628.
    https://doi.org/10.1038/nmeth.1226

    Nazario-Toole, A.E. & Wu, L.P. (2017) Phagocytosis in Insect Immunity. In: Ligoxygakis, P. (Ed.) Advances in Insect Physiology. Academic Press, Cambridge, Massachusetts, pp. 35–82.
    https://doi.org/10.1016/bs.aiip.2016.12.001

    Patnaik, B.B., Patnaik, H.H., Seo, G.W., Jo, Y.H., Lee, Y.S., Lee, B.L. & Han, Y.S. (2014) Gene structure, cDNA characterization and RNAi-based functional analysis of a myeloid differentiation factor 88 homolog in Tenebrio molitor larvae exposed to Staphylococcus aureus infection. Developmental & Comparative Immunology, 46 (2), 208–221.
    https://doi.org/10.1016/j.dci.2014.04.009

    Pili-Floury, S., Leulier, F., Takahashi, K., Saigo, K., Samain, E., Ueda, R. & Lemaitre, B. (2004) In Vivo RNA Interference Analysis Reveals an Unexpected Role for GNBP1 in the Defense against Gram-positive Bacterial Infection in Drosophila Adults. Journal of Biological Chemistry, 279 (13), 12848–12853.
    https://doi.org/10.1074/jbc.M313324200

    Qian, C., Wang, F., Zhu, B.-J., Wei, G.-Q., Li, S., Liu, C.-L. & Wang, L. (2016) Identification and characterization of an Apolipophorin-III gene from Actias selene Hübner (Lepidoptera: Saturniidae). Journal of Asia-Pacific Entomology, 19 (1), 103–108.
    https://doi.org/10.1016/j.aspen.2015.12.008

    Qian, C., Wang, F., Zhu, B.-J., Wei, G.-Q., Sun, Y., Li, S., Wang, L. & Liu, C.-L. (2015) Identification and expression analysis of a novel defense gene from Actias selene. Oriental Insects, 49 (3–4), 264–274.
    https://doi.org/10.1080/00305316.2015.1081648

    Qian, C., Wang, F., Zhu, B., Wang, L., Wei, G., Sun, Y., Li, S. & Liu, C. (2017a) Identification of a hemolin protein from Actias selene mediates immune response to pathogens. International Immunopharmacology, 42, 74–80.
    https://doi.org/10.1016/j.intimp.2016.11.020

    Qian, C., Wang, F., Zhu, B., Wang, L., Wei, G., Sun, Y., Li, S. & Liu, C. (2017b) Identification of a hemolin protein from Actias selene mediates immune response to pathogens. Int Immunopharmacol, 42, 74–80.
    https://doi.org/10.1016/j.intimp.2016.11.020

    Romualdi, C., Bortoluzzi, S., D'Alessi, F. & Danieli, G.A. (2003) IDEG6: a web tool for detection of differentially expressed genes in multiple tag sampling experiments. Physiological Genomics, 12 (2), 159–162.
    https://doi.org/10.1152/physiolgenomics.00096.2002

    Royet, J., Reichhart, J.M. & Hoffmann, J.A. (2005) Sensing and signaling during infection in Drosophila. Current Opinion Immunology, 17 (1), 11–17.
    https://doi.org/10.1016/j.coi.2004.12.002

    Schofield, A.V. & Bernard, O. (2013) Rho-associated coiled-coil kinase (ROCK) signaling and disease. Critical Reviews in Biochemistry and Molecular Biology, 48 (4), 301–316.
    https://doi.org/10.3109/10409238.2013.786671

    Stanley, D. (2006) Prostaglandins and other eicosanoids in insects: biological significance. Annual Review of Entomology, 51, 25–44.
    https://doi.org/10.1146/annurev.ento.51.110104.151021

    Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28 (10), 2731–2739.
    https://doi.org/10.1093/molbev/msr121

    Tang, Q.Y. & Zhang, C.X. (2013) Data Processing System (DPS) software with experimental design, statistical analysis and data mining developed for use in entomological research. Insect Scencei, 20 (2), 254–260.
    https://doi.org/10.1111/j.1744-7917.2012.01519.x

    Tauszig-Delamasure, S., Bilak, H., Capovilla, M., Hoffmann, J.A. & Imler, J.L. (2002) Drosophila MyD88 is required for the response to fungal and Gram-positive bacterial infections. Nature Immunology, 3 (1), 91–97.
    https://doi.org/10.1038/ni747

    Thompson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F. & Higgins, D.G. (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 25 (24), 4876–4882.

    https://doi.org/10.1093/nar/25.24.4876

    Vabulas, R.M., Braedel, S., Hilf, N., Singh-Jasuja, H., Herter, S., Ahmad-Nejad, P., Kirschning, C.J., Da Costa, C., Rammensee, H.G., Wagner, H. & Schild, H. (2002) The endoplasmic reticulum-resident heat shock protein Gp96 activates dendritic cells via the Toll-like receptor 2/4 pathway. Journal of Biological Chemistry, 277 (23), 20847–20853.
    https://doi.org/10.1074/jbc.M200425200

    Wang, L., Feng, Z., Wang, X., Wang, X. & Zhang, X. (2010) DEGseq: an R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics, 26 (1), 136–138.
    https://doi.org/10.1093/bioinformatics/btp612

    Wang, L., Li, Z., Du, C., Chen, W. & Pang, Y. (2007) Characterization and expression of a cecropin-like gene from Helicoverpa armigera. Comparative Biochemistry & Physiology Part B, 148 (4), 417–425.
    https://doi.org/10.1016/j.cbpb.2007.07.010

    Wang, X.Y., Yu, H.Z., Geng, L., Xu, J.P., Yu, D., Zhang, S.Z., Ma, Y. & Fei, D.Q. (2016) Comparative Transcriptome Analysis of Bombyx mori (Lepidoptera) Larval Midgut Response to BmNPV in Susceptible and Near-Isogenic Resistant Strains. PLOS ONE, 11 (5), e0155341.
    https://doi.org/10.1371/journal.pone.0155341

    Wei, R.Y., Bai, J., Zhao, M.F., Xu, B., Li, W.J., Wei, F.X., Xi, Y.Y. & Li, S.Y. (2017) Anti-inflammatory activity of cecropin-A2 from Musca domestica. Microbial Pathogenesis, 110, 637–644.
    https://doi.org/10.1016/j.micpath.2017.07.032

    Xu, Y., Zhou, W., Zhou, Y., Wu, J. & Zhou, X. (2012) Transcriptome and comparative gene expression analysis of Sogatella furcifera (Horvath) in response to southern rice black-streaked dwarf virus. PLoS One, 7 (4), e36238.
    https://doi.org/10.1371/journal.pone.0036238

    Yu, H.-Z., Wang, J., Zhang, S.-Z., Toufeeq, S., Li, B., Li, Z., Yang, L.-A., Hu, P. & Xu, J.-P. (2018a) Molecular characterisation of Apolipophorin-III gene in Samia cynthia ricini and its roles in response to bacterial infection. Journal of Invertebrate Pathology, 159, 61–70.
    https://doi.org/10.1016/j.jip.2018.10.009

    Yu, H.-Z., Wang, J., Zhang, S.-Z., Toufeeq, S., Li, B., Li, Z., Yang, L.-A., Hu, P. & Xu, J.-P. (2018b) Molecular characterisation of Apolipophorin-III gene in Samia cynthia ricini and its roles in response to bacterial infection. Journal of Invertebrate Pathology, 159, 61–70.
    https://doi.org/10.1016/j.jip.2018.10.009

    Zhang, C., Wang, L., Wei, G., Qian, C., Dai, L., Sun, Y., Zhu, B.J. & Liu, C.L. (2015) Small Heat Shock Proteins and Eicosanoid Pathways Modulate Caspase-1 Activity in the Fat Bodies of Antheraea pernyi. bioRxiv. [published online]
    https://doi.org/10.1101/026039

    Zhang, C., Zhu, B., Dai, L.S., Liu, C. & Luo, X. (2016) Nucleopolyhedroviruses (NPV) induce the expression of small heat shock protein 25.4 in Antheraea pernyi. Gene, 591 (2), 327–332.
    https://doi.org/10.1016/j.gene.2016.06.004

    Zhang, N., Zhang, H.J., Zhao, B., Sun, Q.Q., Cao, Y.Y., Li, R., Wu, X.X., Weeda, S., Li, L., Ren, S., Reiter, R.J. & Guo, Y.D. (2014) The RNA-seq approach to discriminate gene expression profiles in response to melatonin on cucumber lateral root formation. Journal of Pineal Research, 56 (1), 39–50.
    https://doi.org/10.1111/jpi.12095

    Zhang, S.Z., Yu, H.Z., Deng, M.J., Ma, Y., Fei, D.Q., Wang, J., Li, Z., Meng, Y. & Xu, J.P. (2018) Comparative transcriptome analysis reveals significant metabolic alterations in eri-silkworm (Samia cynthia ricini) haemolymph in response to 1-deoxynojirimycin. PLoS ONE, 13 (1), e0191080.
    https://doi.org/10.1371/journal.pone.0191080

    Zhao, S., Wang, X., Cai, S., Zhang, S., Luo, H., Wu, C., Zhang, R. & Zhang, J. (2018) A novel peptidoglycan recognition protein involved in the prophenoloxidase activation system and antimicrobial peptide production in Antheraea pernyi. Developmental & Comparative Immunology, 86, 78–85.
    https://doi.org/10.1016/j.dci.2018.04.009