二化螟幼虫热胁迫响应的转录组分析

doi:10.3969/j.issn.1004-1524.2020.05.13

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (5): 849-857.DOI: 10.3969/j.issn.1004-1524.2020.05.13

二化螟幼虫热胁迫响应的转录组分析

朱宇, 刘洋

温州科技职业学院,浙江温州 325006

收稿日期:2019-10-21 出版日期:2020-05-25 发布日期:2020-05-29
作者简介:朱宇(1989—),男,湖北荆州人,博士,讲师,研究方向为昆虫生理生化与分子生物学。E-mail: zhuyuphd@foxmail.com
基金资助:
浙江省教育厅一般科研项目(Y201840543)

Transcriptome analysis on heat tolerance of Chilo suppressalis larvae

ZHU Yu, LIU Yang

Wenzhou Vocational College of Science and Technology, Wenzhou 325006, China

Received:2019-10-21 Online:2020-05-25 Published:2020-05-29

摘要/Abstract

摘要： 二化螟(Chilo suppressalis)对温度的适应能力很强,在高温环境下,二化螟幼虫体内能产生热激蛋白、超氧化物歧化酶和过氧化氢酶等抗氧化物质来消除氧化应激反应对其身体的伤害。本研究旨在通过高通量测序技术对分别置于38 ℃和26 ℃中2 h的二化螟幼虫进行比较转录组学分析,从转录组水平揭示短时高温胁迫对二化螟幼虫的影响。分析结果显示,所有样品的转录组序列比对到参考基因组上的序列占总体的百分比均超过89%,在参考基因组中有唯一比对位置的测序序列占总体的百分比均高于82%,有多个比对位置的序列占总体的百分比均不超过9%。通过差异表达分析得到387个差异表达基因,其中,热激蛋白和细胞色素P450这两类与二化螟热胁迫响应相关的基因为上调表达。对上调的差异基因进行KEGG富集分析,发现内质网内蛋白质加工、寿命调节通路-多物种、植物病原体相互作用和抗原处理及呈现这4条通路为显著富集。本研究揭示了二化螟幼虫热胁迫响应相关基因的总体表达特征,同时可为二化螟热胁迫响应的相关试验提供参考。

关键词: 二化螟, 幼虫, 热胁迫, 转录组

Abstract: Chilo suppressalis has strong adaptability to temperature. In high temperature environments, C. suppressalis larvae can produce antioxidant substances such as heat shock proteins, superoxide dismutase and catalase to eliminate the damage caused by oxidative stress. To reveal the effect of short-term high temperature on C. suppressalis larvae, a comparative transcriptome analysis between two sets of C. suppressalis larvae, treated at 38 ℃ and 26 ℃ for two hours respectively, was performed. The transcriptome results showed that the clean reads of all samples mapped to the reference genome accounted for more than 89%, the percentage of the sequences with unique mapped was higher than 82%, and the percentage of the sequences with multiple positions did not exceed 9%. Through differential expression analysis, 387 differentially expressed genes were obtained. Among them, heat shock protein and cytochrome P450 were up-regulated expression of genes related to the response to heat stress. KEGG enrichment analysis was performed on the up-regulated differentially expressed genes, and the four pathways in the protein processing in endoplasmic reticulum, longevity regulating pathway-multiple species, plant-pathogen interaction and antigen processing and presentation were significantly enriched. This study revealed the overall expression levels of genes involved in heat stress response in the larvae of C. suppressalis, and provided references for related experiments on heat stress response of C. suppressalis.

Key words: Chilo suppressalis, larvae, heat stress, transcriptome

中图分类号:

S433.4

朱宇, 刘洋. 二化螟幼虫热胁迫响应的转录组分析[J]. 浙江农业学报, 2020, 32(5): 849-857.

ZHU Yu, LIU Yang. Transcriptome analysis on heat tolerance of Chilo suppressalis larvae[J]. , 2020, 32(5): 849-857.

参考文献

[1] BALE J S, MASTERS G J, HODKINSON I D, et al.Herbivory in global climate change research: direct effects of rising temperature on insect herbivores[J]. Global Change Biology, 2002, 8(1): 1-16.
[2] 盛承发, 王红托, 盛世余, 等. 我国稻螟灾害的现状及损失估计[J]. 昆虫知识, 2003, 40(4): 289-294.
SHENG C F, WANG H T, SHENG S Y, et al.Pest status and loss assessment of crop damage caused by the rice borers, Chilo suppressalis and Tryporyza incertulas in China[J]. Entomological Knowledge, 2003, 40(4): 289-294.(in Chinese with English abstract)
[3] GUO K, SUN O J, KANG L.The responses of insects to global warming[M]//Recent advances in entomological research. Berlin, Heidelberg: Springer Berlin Heidelberg, 2011: 201-212.
[4] ESTAY S A, LIMA M, BOZINOVIC F.The role of temperature variability on insect performance and population dynamics in a warming world[J]. Oikos, 2014, 123(2): 131-140.
[5] BÜRGI L P, MILLS N J. Ecologically relevant measures of the physiological tolerance of light brown apple moth, Epiphyas postvittana, to high temperature extremes[J]. Journal of Insect Physiology, 2012, 58(9): 1184-1191.
[6] ZHANG S Z, FU W Y, LI N, et al.Antioxidant responses of Propylaea japonica(Coleoptera: Coccinellidae) exposed to high temperature stress[J]. Journal of Insect Physiology, 2015, 73: 47-52.
[7] ANDREW N R,TERBLANCHE J S.The response of insects to climate change[J]. Journal of the Society for Industrial & Applied Mathematics, 2013, 4(2): 1-19.
[8] YANG L H, HUANG H, WANG J J.Antioxidant responses of citrus red mite, Panonychus citri (McGregor) (Acari: Tetranychidae), exposed to thermal stress[J]. Journal of Insect Physiology, 2010, 56(12): 1871-1876.
[9] CHEN H S, SOLANGI G S, GUO J Y, et al.Antioxidant responses of ragweed leaf beetle Ophraella communa(Coleoptera: Chrysomelidae) exposed to thermal stress[J]. Frontiers in Physiology, 2018, 9: 808.
[10] ATAPOUR M, MOHARRAMIPOUR S.Changes of cold hardiness, supercooling capacity, and major cryoprotectants in overwintering larvae of Chilo suppressalis(Lepidoptera: Pyralidae)[J]. Environmental Entomology, 2009, 38(1): 260-265.
[11] 罗举, 张孝羲, 翟保平, 等. 高温对二化螟实验种群生长、存活和繁殖的影响[J]. 生态学报, 2005, 25(4): 931-936.
LUO J, ZHANG X X, ZHAI B P, et al.Effect of high temperature on the growth, survival and reproduction of a laboratory population of the rice stem borer Chilo suppressalis Walker[J]. Acta Ecologica Sinica, 2005, 25(4): 931-936.(in Chinese with English abstract)
[12] RAHMAN M T, KHALEQUZZAMAN M.Temperature requirements for the development and survival of rice stemborers in laboratory conditions[J]. Insect Science, 2004, 11(1): 47-60.
[13] CUI Y D, DU Y Z, LU M X, et al.Antioxidant responses of Chilo suppressalis (Lepidoptera: Pyralidae) larvae exposed to thermal stress[J]. Journal of Thermal Biology, 2011, 36(5): 292-297.
[14] PAN D D, LU M X, LI Q Y, et al.Characteristics and expression of genes encoding two small heat shock protein genes lacking introns from Chilo suppressalis[J]. Cell Stress and Chaperones, 2018, 23(1): 55-64.
[15] CUI Y D, DU Y Z, LU M X, et al.Cloning of the heat shock protein 60 gene from the stem borer, Chilo suppressalis, and analysis of expression characteristics under heat stress[J]. Journal of Insect Science, 2010, 10(100): 1-13.
[16] LU M X, LIU Z X, CUI Y D, et al.Expression patterns of three heat shock proteins in Chilo suppressalis(Lepidoptera: Pyralidae)[J]. Annals of the Entomological Society of America, 2014, 107(3): 667-673.
[17] LU M X, HUA J, CUI Y D, et al.Five small heat shock protein genes from Chilo suppressalis: characteristics of gene, genomic organization, structural analysis, and transcription profiles[J]. Cell Stress and Chaperones, 2014, 19(1): 91-104.
[18] 崔亚东, 杜予州, 陆明星, 等. 热胁迫对二化螟幼虫血淋巴细胞内活性氧、HSP90及细胞凋亡的影响[J]. 昆虫学报, 2010, 53(7): 721-726.
CUI Y D, DU Y Z, LU M X, et al.Effect of thermal stress on the generation of ROS, HSP90 and apoptosis in haemocytes of Chilo suppressalis (Lepidoptera: Pyralidae) larvae[J]. Acta Entomologica Sinica, 2010, 53(7): 721-726.(in Chinese with English abstract)
[19] QI Y X, LIU Y B, RONG W H.RNA-Seq and its applications: a new technology for transcriptomics[J]. Hereditas (Beijing), 2011, 33(11): 1191-1202.
[20] MUTZ K O, HEILKENBRINKER A, LÖNNE M, et al. Transcriptome analysis using next-generation sequencing[J]. Current Opinion in Biotechnology, 2013, 24(1): 22-30.
[21] XIONG Y, LIU X Q, XIAO P A, et al.Comparative transcriptome analysis reveals differentially expressed genes in the Asian Citrus psyllid (Diaphorina citri) upon heat shock[J]. Comparative Biochemistry and Physiology Part D: Genomics and Proteomics, 2019, 30: 256-261.
[22] LI B L, LI M M, WU J X, et al.Transcriptomic analysis of differentially expressed genes in the oriental armyworm Mythimna separata Walker at different temperatures[J]. Comparative Biochemistry and Physiology Part D: Genomics and Proteomics, 2019, 30: 186-195.
[23] LIU Y C, SU H, LI R Q, et al.Comparative transcriptome analysis of Glyphodes pyloalis Walker (Lepidoptera: Pyralidae) reveals novel insights into heat stress tolerance in insects[J]. BMC Genomics, 2017, 18: 974.
[24] 孟凤霞, 吴孔明, 高希武. 利用茭白、荸荠及水稻饲养二化螟的技术研究[J]. 昆虫知识, 2003, 40(5): 469-472.
MENG F X, WU K M, GAO X W.Evaluation of few-flower wildrice, chufa and rice as foods for rearing the striped stem borer, Chilo supperssalis[J]. Entomological Knowledge, 2003, 40(5): 469-472.(in Chinese with English abstract)
[25] BOLGER A M, LOHSE M, USADEL B.Trimmomatic: a flexible trimmer for Illumina sequence data[J]. Bioinformatics, 2014, 30(15): 2114-2120.
[26] ANDREWS S. FastQC: a quality control tool for high throughput sequence data[EB/OL].(2010)[2019-05-14]. http://www.bioinformatics.babraham.ac.uk/projects/fastqc.
[27] PERTEA M, PERTEA G M, ANTONESCU C M, et al.StringTie enables improved reconstruction of a transcriptome from RNA-seq reads[J]. Nature Biotechnology, 2015, 33(3): 290-295.
[28] KIM D, LANGMEAD B, SALZBERG S L.HISAT: a fast spliced aligner with low memory requirements[J]. Nature Methods, 2015, 12(4): 357-360.
[29] LOVE M I, HUBER W, ANDERS S.Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2[J]. Genome Biology, 2014, 15(12): 550.
[30] BENJAMINI Y, YEKUTIELI D.The control of the false discovery rate in multiple testing under dependency[J]. The Annals of Statistics, 2001, 29(4):1165-1188.
[31] JOHNSON M, ZARETSKAYA I, RAYTSELIS Y, et al.NCBI BLAST: a better web interface[J]. Nucleic Acids Research, 2008, 36(Web Server): W5-W9.
[32] KANEHISA M.KEGG: Kyoto encyclopedia of genes and genomes[J]. Nucleic Acids Research, 2000, 28(1): 27-30.
[33] ASHBURNER M, BALL C A, BLAKE J A, et al.Gene Ontology: tool for the unification of biology[J]. Nature Genetics, 2000, 25(1): 25-29.
[34] KING A M, MACRAE T H.Insect heat shock proteins during stress and diapause[J]. Annual Review of Entomology, 2015, 60(1): 59-75.
[35] CHEN H, XU X L, LI Y P, et al.Characterization of heat shock protein 90, 70 and their transcriptional expression patterns on high temperature in adult of Grapholita molesta(Busck)[J]. Insect Science, 2014, 21(4): 439-448.
[36] JOPLIN K H, YOCUM G D, DENLINGER D L.Cold shock elicits expression of heat shock proteins in the flesh fly, Sarcophaga crassipalpis[J]. Journal of Insect Physiology, 1990, 36(11): 825-834.
[37] CHAPUIS M P, SIMPSON S J, BLONDIN L, et al.Taxa-specific heat shock proteins are over-expressed with crowding in the Australian plague locust[J]. Journal of Insect Physiology, 2011, 57(11): 1562-1567.
[38] MICHAUD M R, TEETS N M, PEYTON J T, et al.Heat shock response to hypoxia and its attenuation during recovery in the flesh fly, Sarcophaga crassipalpis[J]. Journal of Insect Physiology, 2011, 57(1): 203-210.
[39] WANG H H, REITZ S R, WANG L X, et al.The mRNA expression profiles of five heat shock protein genes from Frankliniella occidentalis at different stages and their responses to temperatures and insecticides[J]. Journal of Integrative Agriculture, 2014, 13(10): 2196-2210.
[40] CHEN B, KAYUKAWA T, MONTEIRO A, et al.The expression of the HSP90 gene in response to winter and summer diapauses and thermal-stress in the onion maggot, Delia antiqua[J]. Insect Molecular Biology, 2005, 14(6): 697-702.
[41] RINEHART J P, YOCUM G D, DENLINGER D L.Developmental upregulation of inducible hsp70 transcripts, but not the cognate form, during pupal diapause in the flesh fly, Sarcophaga crassipalpis[J]. Insect Biochemistry and Molecular Biology, 2000, 30(6): 515-521.
[42] CHENG W N, LI D, WANG Y, et al.Cloning of heat shock protein genes (hsp70, hsc70 and hsp90) and their expression in response to larval diapause and thermal stress in the wheat blossom midge, Sitodiplosis mosellana[J]. Journal of Insect Physiology, 2016, 95: 66-77.
[43] GARRIDO C, PAUL C, SEIGNEURIC R, et al.The small heat shock proteins family: The long forgotten chaperones[J]. The International Journal of Biochemistry & Cell Biology, 2012, 44(10): 1588-1592.
[44] KIRTON S B, MURRAY C W, VERDONK M L, et al.Prediction of binding modes for ligands in the cytochromes P450 and other heme-containing proteins[J]. Proteins: Structure, Function, and Bioinformatics, 2005, 58(4): 836-844.
[45] FEYEREISEN R.Evolution of insect P450[J]. Biochemical Society Transactions, 2006, 34(6): 1252-1255.
[46] IGA M, KATAOKA H.Recent studies on insect hormone metabolic pathways mediated by cytochrome P450 enzymes[J]. Biological and Pharmaceutical Bulletin, 2012, 35(6): 838-843.
[47] GIRAUDO M, HILLIOU F, FRICAUX T, et al.Cytochrome P450s from the fall armyworm (Spodoptera frugiperda): responses to plant allelochemicals and pesticides[J]. Insect Molecular Biology, 2015, 24(1): 115-128.
[48] POUPARDIN R, RIAZ M A, VONTAS J, et al.Transcription profiling of eleven cytochrome P450s potentially involved in xenobiotic metabolism in the mosquito Aedes aegypti[J]. Insect Molecular Biology, 2010, 19(2): 185-193.
[49] ZHU M, ZHANG W X, LIU F, et al.Characterization of an Apis cerana cerana cytochrome P450 gene (AccCYP336A1) and its roles in oxidative stresses responses[J]. Gene, 2016, 584(2): 120-128.
[50] 刘井兰, 于建飞, 吴进才, 等. 昆虫活性氧代谢[J]. 昆虫知识, 2006, 43(6): 752-756.
LIU J L, YU J F, WU J C, et al.Insect reactive oxygen metabolization[J]. Chinese Bulletin of Entomology, 2006, 43(6): 752-756.(in Chinese with English abstract)

编辑推荐

Metrics

阅读次数

全文

1656

HTML			PDF

最新录用	在线预览	正式出版	最新录用	在线预览	正式出版
0	0	5	0	0	1651

来源	本网站	其他网站

次数	1126	530
比例	68%	32%

摘要

808

最新录用	在线预览	正式出版

0	0	808

来源	本网站	其他网站

次数	807	1
比例	100%	0%

二化螟幼虫热胁迫响应的转录组分析

Transcriptome analysis on heat tolerance of Chilo suppressalis larvae

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	尹明华, 曹晴, 陈红, 邓思宇, 邓燕梅. 江西铅山红芽芋和青秆芋的转录组比较分析[J]. 浙江农业学报, 2020, 32(9): 1533-1543.
[2]	刘新雨, 田洁. 大蒜转录组简单重复序列标记分析与分子标记开发[J]. 浙江农业学报, 2020, 32(9): 1615-1625.
[3]	葛金涛, 王江英, 赵文静, 邵小斌, 朱朋波, 汤雪燕, 孙明伟, 刘兴满. 魏可葡萄气生根发育的转录组分析[J]. 浙江农业学报, 2020, 32(9): 1645-1655.
[4]	朱晓林, 魏小红, 王宝强, 王贤, 张明君. c-GMP诱导对盐胁迫下番茄的转录组分析[J]. 浙江农业学报, 2020, 32(10): 1788-1797.
[5]	王其, 陈小洁, 顾双月, 张欣悦, 杭天露, 丁婷. 杜仲内生拮抗细菌DZSY21诱导玉米抗病基因表达变化的转录组学研究[J]. 浙江农业学报, 2019, 31(3): 345-354.
[6]	董捷, 郭蕊, 郭海坤, 李君竹, 方勐鸽, 王德前. 浙江省淳安县中华蜜蜂囊状幼虫病流行病学调查[J]. 浙江农业学报, 2019, 31(3): 365-371.
[7]	王华, 汪王微, 王冬良, 张石虎, 胡新芳, 卢诗雨, 龚雪梅. 杜鹃花叶片转录组测序数据组装及功能注释[J]. 浙江农业学报, 2018, 30(7): 1149-1159.
[8]	白琪, 鲁艳辉, 郑许松, 吕仲贤. 二化螟两种P450基因CYP4M38和CYP4M39的时空表达分析[J]. 浙江农业学报, 2018, 30(4): 521-527.
[9]	解彦玲, 王鸿权, 张璐, 侯志贤, 刀晨, 江亮亮, 熊翠玲, 郑燕珍, 徐细建, 黄枳腱, 郭睿, 陈大福. 中华蜜蜂幼虫肠道的高表达基因分析[J]. 浙江农业学报, 2017, 29(9): 1575-1580.
[10]	杜宇, 熊翠玲, 史秀丽, 郑燕珍, 付中民, 徐细建, 陈大福, 郭睿. 意大利蜜蜂6日龄幼虫肠道内球囊菌的差异表达基因分析[J]. 浙江农业学报, 2017, 29(7): 1119-1128.
[11]	冯琛, 汤浩茹, 江雷雨, 宋霞, 张云婷, 叶云天, 陈清, 孙勃. 红蓝光对草莓转录组特异表达基因密码子使用偏好性的影响[J]. 浙江农业学报, 2017, 29(4): 566-574.
[12]	沈春修. 水稻LOC_Os10g05490位点冷胁迫条件下表达分析及CRISPR/Cas9定向编辑[J]. 浙江农业学报, 2017, 29(2): 177-185.
[13]	洪森荣, 吴夏俊鹏, 徐文慧, 占学林, 谢妮妮, 蒋妍, 汪金华, 凌飞, 吴丽霞, 万琳. 低温离体保存黄独微型块茎转录组、蛋白质组和代谢组的关联分析[J]. 浙江农业学报, 2017, 29(11): 1827-1834.
[14]	裴翠明，张振亚，马进*. 南方型紫花苜蓿叶片盐胁迫应答转录因子鉴定与分析[J]. 浙江农业学报, 2016, 28(4): 550-.
[15]	马婧1，成铁龙1，2，*，孙灿岳3，邓楠1，史胜青1，江泽平1. 草麻黄高通量转录组分析及黄酮类代谢途径相关基因的鉴定[J]. 浙江农业学报, 2016, 28(4): 609-.