Please wait a minute...
Journal of Integrative Agriculture  2018, Vol. 17 Issue (05): 1012-1022    DOI: 10.1016/S2095-3119(17)61737-4
Special Focus: Insect heat shock proteins and their underlying functions Advanced Online Publication | Current Issue | Archive | Adv Search |
Genes encoding heat shock proteins in the endoparasitoid wasp, Cotesia chilonis, and their expression in response to temperatures
PAN Dan-dan1, CAO Shuang-shuang1, LU Ming-xing1, HANG San-bao1, DU Yu-zhou1, 2
1 School of Horticulture and Plant Protection & Institute of Applied Entomology, Yangzhou University, Yangzhou 225009, P.R.China
2 Joint International Research Laboratory of Agriculture and Agri-Product Safety, Ministry of Education/Yangzhou University, Yangzhou 225009, P.R.China
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
Abstract  Five genes encoding heat shock proteins (HSPs), Cchsp40, Cchsp60, Cchsp70, Cchsc70 and Cchsp90, were cloned and sequenced from Cotesia chilonis using RT-PCR and RACE.  The cDNA sequences of Cchsp40, Cchsp60, Cchsp70, Cchsc70 and Cchsp90 were 1 265, 2 551, 2 094, 2 297 and 2 635 bp in length, respectively, with a molecular weight (MW) of 39.1, 60.6, 71.45, 70.19 and 82.92 kDa, respectively.  The predicted amino acid sequences of these proteins showed high similarities with published HSPs of other insects in Hymenoptera.  Analysis of genomic DNAs indicated that Cchsp40, Cchsp60, Cchsp70, Cchsc70 and Cchsp90 lacked introns, but Cchsc70 contained an intron.  The results also suggested that CcHSP40 in C. chilonis was the Type II HSP40, CcHSP60 was a member of the mitochondrial HSP60 family, and CcHSP90 was a part of cytoplasmic HSP90A family.  Expression patterns varied in the five Cchsps in response to temperature.  Expression of Cchsp40 and Cchsp60 was induced significantly by cold but not heat stress.  Cchsp70 and Cchsc70 showed similar response to the thermal stress and could be induced by both cold and heat, but their expression levels were consistently lower than that of Cchsp40 and Cchsp60Cchsp90 could be induced by heat stress and mild cold, but not cold stress.  In addition, the results demonstrated Cchsc70 might be constitutive and inducible protein that was expressed during normal cell functioning and also up-regulated in response to stressful stimuli while Cchsp70 was solely inducible protein induced by temperature changes.  Overall, results generated from this study could significantly advance the understanding of Cchsps in response to temperature and provide important biological information for C. chilonis insects that reared under different temperatures.  
Keywords:  Cotesia chilonis        HSPs        genomic structure        temperature        expression  
Received: 20 June 2017   Accepted:

This research was funded by the National Key R&D Program of China (2017YFD0200400) and the National Basic Research Program of China (973 Program, 2013CB127604).

Corresponding Authors:  Correspondence DU Yu-zhou, E-mail:   

Cite this article: 

PAN Dan-dan, CAO Shuang-shuang, LU Ming-xing, HANG San-bao, DU Yu-zhou. 2018. Genes encoding heat shock proteins in the endoparasitoid wasp, Cotesia chilonis, and their expression in response to temperatures. Journal of Integrative Agriculture, 17(05): 1012-1022.

Aevermann B D, Waters E R. 2008. A comparative genomic analysis of the small heat shock proteins in Caenorhabditiselegans and briggsae. Genetica, 133, 307–319.

Akerfelt M, Morimoto R I, Sistonen L. 2010. Heat shock factors: Integrators of cell stress, development and lifespan. Nature Reviews Molecular Cell Biology, 11, 545–555.

Boorstein W R, Ziegelhoffer T, Craig E A. 1994. Molecular evolution of the HSP70 multigene family. Journal of Molecular Evolution, 38, 1–17.

Boutet I, Tanguy A, Rousseau S, Auffret M, Moraga D. 2003. Molecular identification and expression of heat shock cognate 70 (hsc70) and heats shock protein 70 (hsp70) genes in the Pacific oyster Crassostrea gigas. Cell Stress and Chaperones, 8, 76–85.

Cahan S H, Nguyen A D, Stanton-Geddes J, Penick C A, Hernáiz-Hernández Y, DeMarco B B, Gotelli N J. 2017. Modulation of the heat shock response is associated with acclimation to novel temperatures but not adaptation to climatic variation in the ants Aphaenogaster picea and

A. rudis. Comparative Biochemistry and Physiology (Part A: Molecular and Integrative Physiology), 204, 113–120.

Cajo G C, Horne B E, Kelley W L, Schwager F, Georgopoulos C, Genevaux P. 2006. The role of the DIF motif of the DnaJ (Hsp40) co-chaperone in the regulation of the Dnak (Hsp70) chaperone cycle. Journal of Biological Chemistry, 281, 12436–12444.

Callahan M K, Chaillot D, Jacquin C, Clark P R, Menoret A. 2002. Differential acquisition of antigenic peptides by Hsp70 and Hsc70 under oxidative conditions. Journal of Biological Chemistry, 277, 33604–33609.

Caplan A J, Cyr D M, Douglas M G. 1993. Eukaryotic homologs of Escherichia coli DnaJ: A diverse protein family that functions with Hsp70 stress proteins. Journal of Molecular Biology, 4, 555–563.

Chen B, Zhong D, Monteiro A. 2006. Comparative genomics and evolution of the HSP90 family of genes across all kingdoms of organisms. BMC Genomics, 7, 156.

Chen H, Xu X, Li Y, Wu J. 2014. Characterization of heat shock protein 90, 70 and their transcriptional expression patterns on high temperature in adult of Grapholita molesta (Busck). Insect Science, 21, 439–448.

Chen H C, Lou Y G, Cheng J A. 2002. Research and application of Apanteles chilonis, a parasitoid of rice striped stemborer Chilo suppressalis. Chinese Journal of Biological Control, 18, 90–93. (in Chinese)

Comeron J M. 2004. Selective and mutational patterns associated with gene expression in humans: Influences on synonymous composition and intron presence. Genetics, 167, 1293–1304.

Cui Y D, Du Y Z, Lu M X, Qiang C K. 2014. Cloning of the heat shock protein 60 gene from the stem borer, Chilo suppressalis, and analysis of expression characteristics under heat stress. Journal of Insect Science, 10, 100.

Daugaard M, Rohde M, Jaattela M. 2007. The heat shock protein 70 family: Highly homologous proteins with overlapping and distinct functions. Federation of European Biochemical Societies, 581, 3702–3710.

Fan C Y, Lee S, Ren H Y, Cyr D M. 2004. Exchangeable chaperone modules contribute to specification of type I and type II Hsp40 cellular function. Molecular and Cellular Biology, 15, 761–773.

Feder M E, Hofmann G E. 1999. Heat-shock proteins, molecular chaperones, and the stress response: Evolutionary and ecological physiology. Annual Review of Physiology, 61, 243–282.

Fink A L. 1999. Chaperone-mediated protein folding. Physiological Reviews, 79, 425–449.

Galichet P F. 1979. Hibernation of Apanteles chilonis Mun. [Hym.: Braconidae] under Mediterranean climate. Entomophaga, 24, 119–130.

Gupta R S. 1995 Phylogenetic analysis of the 90 kD heat-shock family of protein sequences and an examination of the relationship among animals, plants, and fungi species. Molecular Biology and Evolution, 12, 1063–1073.

Hang S B. 1993. Studies on the rearing method of Apanteles chilonis in laboratory. Journal of Biosafety, 2, 42–47. (in Chinese)

Hang S B, Lin G L. 1989. Biological characteristics of Apanteles chilonis (Hymenoptera: Braconidae), a parasite of Chilo suppressalis (Lepidoptera: Pyralidae). Chinese Journal of Biological Control, 5, 16–18. (in Chinese)

Hausmann C, Samietz J, Dorn S. 2005. Thermal orientation of Anthonomus pomorum (Coleoptera: Curculionidae) in early spring. Physiological Entomology, 30, 48–53.

Hoffmann A A, Parsons P A. 1991. Evolutionary genetics and environmental stress. Journal of Evolutionary Biology, 7, 634–635.

Huang J, Wu S F, Ye G Y. 2011. Evaluation of lethal effects of chlorantraniliprole on Chilo suppressalis and its larval parasitoid, Cotesia chilonis. Chinese Journal of Eco-Agriculture, 10, 1134–1138. (in Chinese)

Jindal S, Dudani A K, Singh B, Harley C B, Gupta R S. 1989. Primary structure of a human mitochondrial protein homologous to the bacterial and plant chaperonins and to the 65-kilodalton mycobacterial antigen. Molecular and Cellular Biology, 9, 2279–2283.

Jing X H, Kang L. 2004. Overview and evaluation of research methodology for insect cold hardiness. Entomological Knowledge, 40, 7–10. (in Chinese)

Kim K K, Kim R, Kim S H. 1998. Crystal structure of a small heat shock protein. Nature, 394, 595–599.

King, A M, Macrae T H. 2014. Insect heat shock proteins during stress and diapause. Annual Review of Entomology, 60, 59.

Kumar S, Stecher G, Tamura K. 2016. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33, 1870–1874.

Li H B, Du Y Z. 2013. Molecular cloning and characterization of an hsp90/70 organizing protein gene from Frankliniella occidentalis. Gene, 520, 148–155.

Li Z, Srivastava P. 2004. Heat-shock proteins. Current Protocols in Immunology, A-1T.

Liberek K, Georgopoulos C, Zylicz M. 1988. Role of the Escherichia coli DnaK and DnaJ heat shock proteins in the initiation of bacteriophage lambda DNA replication. Proceedings of the National Academy of Sciences of the United States of America, 85, 6632–6636.

Lu M X, Cao S S, Du Y Z, Liu Z X, Liu P Y, Li J Y. 2013. Diapause, signal and molecular characteristics of overwintering Chilo suppressalis (Insecta: Lepidoptera: Pyralidae). Scientific Reports, 3, 3211.

Lu M X, Li H B, Zheng Y T, Shi L, Du Y Z. 2016. Identification, genomic organization and expression profiles of four heat shock protein genes in the Western flower thrips, Frankliniella occidentalis. Journal of Thermal Biology, 57, 110–118.

Mahroof R, Zhu K Y, Neven L, Subramanyam B, Bai J. 2005. Expression patterns of three heat shock protein 70 genes among developmental stages of the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae). Comparative Biochemistry and Physiology (Part A: Molecular & Integrative Physiology), 141, 247–256.

McKay D B, Wilbanks S M, Flaherty K M, Ha J H, O’Brien M C, Shirvanee L L. 1994. Stress-70 proteins and their interaction with nucleotides. Cold Spring Harbor Monograph Archive, 26, 153–177.

Morimoto R I, Tissieres A, Georgopoulos C. 1990. The stress response, function of the proteins, and perspectives. Cold Spring Harbor Monograph Archive, 19, 1–36.

Padmini E. 2010. Physiological adaptations of stressed fish to polluted environments: Role of heat shock proteins. Reviews of Environmental Contamination and Toxicology Volume, 206, 1–27.

Pallant J. 2007. SPSS survival manual: A step by step guide to data analysis using SPSS for windows (version 12). Open University Press, 37, 597–598.

Pan D D, Liu Z X, Lu M X, Cao S S, Yan W F, Du Y Z. 2016. Species and occurrence dynamics of parasitic wasps of the rice stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae) in Yangzhou. Journal of Environmental Entomology, 38, 1106–1113.

Pan D D, Lu M X, Li Q Y, Du Y Z. 2017. Characteristics and expression of genes encoding two small heat shock protein genes lacking introns from Chilo suppressalis. Cell Stress and Chaperones, 3, 1–10.

Qiu X B, Shao Y, Miao S, Wang L. 2006. The diversity of the DnaJ/Hsp40 family, the crucial partners for Hsp70 chaperones. Cellular and Molecular Life Sciences, 63, 2560–2570.

Ravaux J, Toullec J Y, Leger N, Lopez P, Gaill F, Shillito B. 2007. First hsp70 from two hydrothermal vent shrimps, Mirocaris fortunate and Rimicaris exoculata: Characterization and sequence analysis. Gene, 386, 162–172.

Rinehart J P, Li A, Yocum G D, Robich R M, Hayward S A, Denlinger D L. 2007. Up-regulation of heat shock proteins is essential for cold survival during insect diapause. Proceedings of the National Academy of Sciences of the United States of America, 104, 11130–11137.

Samietz J, Salser M A, Dingle H. 2005. Altitudinal variation in behavioural thermoregulation: Local adaptation vs. plasticity in California grasshoppers. Journal of Evolutionary Biology, 18, 1087–1096.

Sanders B M, Pascoe V M, Nakagawa P A, Martin L S. 1992. Persistence of the heat shock response over time in a common Mytilus mussel. Molecular Marine Biology and Biotechnology, 1, 147–154.

Schmittgen T D, Tamura K J. 2008. Analyzing real-time PCR data by the comparative CT method. Nature Protocols, 3, 1101–1108.

Slavotinek A M, Biesecker L G. 2001. Unfolding the role of chaperones and chaperonins in human disease. Trends in Genetics, 17, 528.

Sonoda S, Ashfaq M, Tsumuki H. 2006. Cloning and nucleotide sequencing of three heat shock protein genes (hsp90, hsc70, and hsp19.5) from the diamondback moth, Plutella xylostella (L.) and their expression in relation to developmental stage and temperature. Archives of Insect Biochemistry and Physiology, 62, 80–90.

Sonoda S, Ashfaq M, Tsumuki H. 2007. A comparison of heat shock protein genes from cultured cells of the Cabbage armyworm, Mamestra brassicae, in response to heavy metals. Archives of Insect Biochemistry and Physiology, 65, 210–222.

Sørensen J G. 2010. Application of heat shock protein expression for detecting natrual adaptation and exposure to stress in natural populations. Current Zoology, 56, 703–713.

Sørensen J G, Kristensen T N, Loeschcke V. 2003. The evolutionary and ecological role of heat shock proteins. Ecology Letters, 6, 1025–1037.

Sun M, Lu M X, Tang X T, Du Y Z. 2014. Molecular cloning and sequence analysis of the HSP83 gene in Sesamia inferens (Walker) (Lepidoptera: Noctuidae). Chinese Journal of Applied Entomology, 51, 1246–1254. (in Chinese)

Tang T, Wu C, Li J, Ren G, Huang D, Liu F. 2012. Stress-induced HSP70 from Musca domestica plays a functionally significant role in the immune system. Journal of Insect Physiology, 58, 1226–1234.

Tang  X T, Sun M, Lu M X, Du Y Z. 2015. Expression patterns of five heat shock proteins in Sesamia inferens (Lepidoptera: Noctuidae) during heat stress. Journal of Asia-Pacific Entomology, 18, 529–533.

Walsh P, Bursac D, Law Y C, Cyr D, Lithgow T. 2004. The J-protein family: Modulating protein assembly, disassembly and translocation. EMBO Reports, 5, 567–571.

Wang H S, Wang X H, Guo W, Zhang S F, Kang L. 2007. cDNA cloning of heat shock proteins and their expression in the two phases of the Migratory locust. Insect Molecular Biology, 16, 207–219.

Wang P, Xu P, Zhou L, Zeng S, Li G. 2017. Molecular cloning, characterization, and expression analysis of HSP60 in mandarin fish Siniperca chuatsi. Israeli Journal of Aquaculture-Bamidgeh, 69, 1–13.

Wang X R, Wang C, Ban F X, Zhu D T, Liu S S, Wang X W. 2017. Genome-wide identification and characterization of HSP gene superfamily in whitefly (Bemisia tabaci) and expression profiling analysis under temperature stress. Insect Science, doi: 10.1111/1744-7917.12505

Waters E R, Aevermann B D, Sanders-Reed Z. 2008. Comparative analysis of the small heat shock proteins in three angiosperm genomes. Cell Stress and Chaperones, 13, 127–142.

Willmer P, Stone G, Johnston I. 2000. Environmental physiology of animals. Blackwell Science, 71, 57–84.

Wu S F, Sun F D, Qi Y X, Yao Y, Fang Q, Huang J, Stanley D, Ye G Y. 2013. Parasitization by Cotesia chilonis influences gene expression in fatbody and hemocytes of Chilo suppressalis. PLoS ONE, 8, e74309.

Xu P J, Xiao J H, Liu L, Li T, Huang D W. 2010. Molecular cloning and characterization of four heat shock protein genes from Macrocentrus cingulum (Hymenoptera: Braconidae). Molecular Biology Reports, 37, 2265–2272.

Xu Q, Zou Q, Zheng H, Zhang F, Tang B, Wang S. 2011. Three heat shock proteins from Spodoptera exigua: Gene cloning, characterization and comparative stress response during heat and cold shocks. Comparative Biochemistry and Physiology (Part B: Biochemistry and Molecular Biology), 159, 92–102.

Yochem J, Uchida H, Sunshine M, Saito H, Georgopoulos C P, Feiss M. 1978. Genetic analysis of two genes, DnaJ and DnaK, necessary for Escherichia coli and Bacteriophage lambda DNA replication. Molecular Genetics and Genomics, 164, 9–14.

Zhang Q R, Denlinger D L. 2010. Molecular characterization of heat shock protein 90, 70 and 70 cognate cDNAs and their expression patterns during thermal stress and pupal diapause in the corn earworm. Journal of Insect Physiology, 56, 138–150.
[1] ZHANG Li-hua, ZHU Ling-cheng, XU Yu, LÜ Long, LI Xing-guo, LI Wen-hui, LIU Wan-da, MA Feng-wang, LI Ming-jun, HAN De-guo. Genome-wide identification and function analysis of the sucrose phosphate synthase MdSPS gene family in apple[J]. >Journal of Integrative Agriculture, 2023, 22(7): 2080-2093.
[2] HOU Qian-dong, HONG Yi, WEN Zhuang, SHANG Chun-qiong, LI Zheng-chun, CAI Xiao-wei, QIAO Guang, WEN Xiao-peng. Molecular characterization of the SAUR gene family in sweet cherry and functional analysis of PavSAUR55 in the process of abscission[J]. >Journal of Integrative Agriculture, 2023, 22(6): 1720-1739.
[3] WANG Ke, HE Yan-yan, ZHANG You-jun, GUO Zhao-jiang, XIE Wen, WU Qing-jun, WANG Shao-li. Characterization of the chemosensory protein EforCSP3 and its potential involvement in host location by Encarsia formosa[J]. >Journal of Integrative Agriculture, 2023, 22(2): 514-525.
[4] LI Zhi-qi, Xie Qian, YAN Jia-hui, CHEN Jian-qing, CHEN Qing-xi. Genome-wide identification and characterization of the abiotic-stress-responsive lipoxygenase gene family in diploid woodland strawberry (Fragaria vesca)[J]. >Journal of Integrative Agriculture, 2022, 21(7): 1982-1996.
[5] CHEN Rong-zhu, SHEN Xu, ZHANG Shu-ting, ZHAO Hua, CHEN Xiao-hui, XU Xiao-ping, HUO Wen, ZHANG Zi-hao, LIN Yu-ling, LAI Zhong-xiong. Genome-wide identification and expression analysis of Argonaute gene family from longan embryogenic callus[J]. >Journal of Integrative Agriculture, 2021, 20(8): 2138-2155.
[6] QU Cheng, WANG Ran, CHE Wu-nan, LI Feng-qi, ZHAO Hai-peng, WEI Yi-yun, LUO Chen, XUE Ming. Identification and tissue distribution of odorant binding protein genes in Harmonia axyridis (Coleoptera: Coccinellidae)[J]. >Journal of Integrative Agriculture, 2021, 20(8): 2204-2213.
[7] ZHOU Nian-bing, ZHANG jun, FANG Shu-liang, WEI Hai-yan, ZHANG Hong-cheng. Effects of temperature and solar radiation on yield of good eating-quality rice in the lower reaches of the Huai River Basin, China[J]. >Journal of Integrative Agriculture, 2021, 20(7): 1762-1774.
[8] YANG Xian-ming, SONG Yi-fei, SUN Xiao-xu, SHEN Xiu-jing, WU Qiu-lin, ZHANG Hao-wen, ZHANG Dan-dan, ZHAO Sheng-yuan, LIANG Ge-mei, WU Kong-ming . Population occurrence of the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), in the winter season of China[J]. >Journal of Integrative Agriculture, 2021, 20(3): 772-782.
[9] BI Yu-lin, YANG Shu-yan, WANG Hai-yan, CHANG Guo-bin, CHEN Guo-hong. Follicle-stimulating hormone is expressed in ovarian follicles of chickens and promotes ovarian granulosa cell proliferation[J]. >Journal of Integrative Agriculture, 2021, 20(10): 2749-2757.
[10] Everlyne M’mbone MULEKE, WANG Yan, ZHANG Wan-ting, XU Liang, YING Jia-li, Bernard K. KARANJA, ZHU Xian-wen, FAN Lian-xue, Zarwali AHMADZAI, LIU Li-wang. Genome-wide identification and expression profiling of MYB transcription factor genes in radish (Raphanus sativus L.)[J]. >Journal of Integrative Agriculture, 2021, 20(1): 120-131.
[11] QIN Jin-xia, JIANG Yu-jie, LU Yun-ze, ZHAO Peng, WU Bing-jin, LI Hong-xia, WANG Yu, XU Sheng-bao, SUN Qi-xin, LIU Zhen-shan. Genome-wide identification and transcriptome profiling reveal great expansion of SWEET gene family and their wide-spread responses to abiotic stress in wheat (Triticum aestivum L.)[J]. >Journal of Integrative Agriculture, 2020, 19(7): 1704-1720.
[12] SONG Jie, LU Ming-xing, DU Yu-zhou. Molecular cloning and expression patterns of two small heat shock proteins from Chilo suppressalis (Walker)[J]. >Journal of Integrative Agriculture, 2020, 19(6): 1522-1529.
[13] WANG Yi-fan, LIAO Yu-qiu, WANG Ya-peng, YANG Jiang-wei, ZHANG Ning, SI Huai-jun. Genome-wide identification and expression analysis of StPP2C gene family in response to multiple stresses in potato (Solanum tuberosum L.)[J]. >Journal of Integrative Agriculture, 2020, 19(6): 1609-1624.
[14] CAO Yue, GAO Zhong-cheng, WU Zheng-chang, WANG Hai-fei, BAO Wen-bin. Tissue-specific expression and correlation with promoter DNA methylation of the LBP gene in pigs[J]. >Journal of Integrative Agriculture, 2020, 19(4): 1055-1064.
[15] XU Jin-bo, ZHANG Cui-ping, WUNIERBIEKE Mei-li, YANG Xiao-fei, LI Yi-lang, CHEN Xiao-bin, CHEN Gong-you, ZOU Li-fang. An improved protein expression system for T3SS genes regulation analysis in Xanthomonas oryzae pv. oryzae[J]. >Journal of Integrative Agriculture, 2019, 18(6): 1189-1198.
No Suggested Reading articles found!