Korean Journal of Ecology and Environment (생태와환경)

The Korean Society of Limnology (한국수생태학회)
권호 표지
DOI QR코드

DOI QR Code

Expression of the Heat Shock Protein 70 Gene and External Developmental Traits of Two Bivalvia Species, Crassostrea gigas and Mytilus galloprovincialis, under Aquaculture Environments

사육환경에 따른 이매패류 (Crassostrea gigas, Mytilus galloprovincialis)의 외부형질 성장과 Heat Shock Protein 70 유전자 발현

  • Kim, Won-Seok (Department of Fisheries and Ocean Science, Chonnam National Unversity) ;
  • Park, Kiyun (Department of Fisheries and Ocean Science, Chonnam National Unversity) ;
  • Kim, Jong Kyu (Department of Fisheries and Ocean Science, Chonnam National Unversity) ;
  • Kwak, Ihn-Sil (Department of Fisheries and Ocean Science, Chonnam National Unversity)
  • 김원석 (전남대학교 해양기술학부) ;
  • 박기연 (전남대학교 해양기술학부) ;
  • 김종규 (전남대학교 해양기술학부) ;
  • 곽인실 (전남대학교 해양기술학부)
  • Received : 2015.11.10
  • Accepted : 2016.02.22
  • Published : 2016.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

Environmental changes exert harmful effects on organisms inhabiting coastal regions. These changes are also associated with reduced production in aquaculture farms. In this study, we investigated internal and external responses of two Bivalvia species (Crassostrea gigas and Mytilus galloprovincialis) in Gamak Bay under stressful environmental conditions in aquaculture farms. We investigated external responses such as weight, size, and environment exposure time, and analyzed the expression of the HSP70 gene. C. gigas HSP70 gene expression level was significantly high in the C3 aquaculture farm site, but the weight and size of C. gigas were high in the C2 aquaculture farm site. The response of C. gigas HSP70 mRNA was associated with the environmental exposure time in each aquaculture farm. Expression of M. galloprovincialis HSP70 gene was found to be significantly higher in the M2 aquaculture farm site than in the M1 site, whereas the weight of M. galloprovincialis was observed to be higher in the M1 site. The size and environmental exposure time of M. galloprovincialis were similar between M1 and M2 sites. In addition, HSP70 sequences of C. gigas and M. galloprovincialis showed high similarity with that of another marine species. According to our results, there were differences in internal responses following environmental stress in aquaculture farms, with respect to HSP70 gene expression. The results suggest that the HSP70 gene is a useful molecular indicator for monitoring stress responses in Bivalvia species in the field.

연안의 다양한 환경변화는 서식 생물에 영향을 미치고, 양식장의 생산량 감소와 연결되고 있는 추세이다. 본 연구에서는 가막만의 대표적인 양식종인 패류 C. gigas와 M. galloprovincialis의 서식환경에 따른 스트레스 정도를 파악하고자 하였다. 이를 위해, 각 종의 체중량, 각장과 각고, 양식장 사육기간을 조사하고, 각 종의 계통학적 HSP 70 sequence를 비교한 후, 각 종의 HSP 70 유전자 발현을 분석하였다. 그 결과, C. gigas의 체중량, 각장과 각고는 C2 양식장이 높게 나타났으나, 양식장 환경 사육기간과 HSP 70 유전자 발현은 C3 양식장이 가장 높았다. M. galloprovincialis는 M1 양식장의 체중량이 높게 나타났으며 각장과 각고, 사육기간은 M2와 유사하였으나, HSP 70 유전자 발현은 M2 양식장이 통계적으로 유의한 수준으로 높게 나타났다. 그리고 C. gigas와 M. galloprovincialis의 HSP 70 sequence 분석을 통해서 다른 해양 종들과 높은 유사성이 있음을 확인하였다. 이 결과는 서식환경에 따라 생물의 외부적 형질뿐만 아니라 내부적 스트레스를 HSP 70 유전자 발현을 통하여 파악할 수 있으며 HSP 70은 외부환경 스트레스를 평가하는 지표 유전자로서 활용할 수 있을 것이다.

Keywords

References

  1. Augustine, S.M., J.A. Narayan, D.P. Syamaladevi, C. Appunu, M. Chakravarthi, V. Ravichandran and M. Subramonian. 2015. Erianthus arundinaceus HSP70 (EaHSP70) overexpression increases drought and salinity tolerance in sugarcane (Saccharum spp. Hybrid). Plant Science 232: 23-34. https://doi.org/10.1016/j.plantsci.2014.12.012
  2. Basile, A., S. Sorbo, M. Lentini, D. Castiglia, P. Cianciullo, B. Conte, S. Loppi and S. Esposito. 2015. Effects of heavy metals on ultrastructure and HSP 70 induction in Lemna minor L. exposed to water along the Sarno River, Italy. Ecotoxicology and Environmental Safety 114: 93-101. https://doi.org/10.1016/j.ecoenv.2015.01.009
  3. Bierkens, J.P.E.G. 2000. Application and pitfalls of stress-proteins in biomonitoring. Toxicology 153: 61-72. https://doi.org/10.1016/S0300-483X(00)00304-8
  4. Carlo, G.A., G. Stefania, M. Massimo, B. Maura, F. Daniele, d'E. Giuseppe, P. Marianna, B. Luca and R. Francesco. 2015. Pollutants bioavailability and toxicology risk from microplastics to marine mussels. Environmental Pollution 198: 211-222. https://doi.org/10.1016/j.envpol.2014.12.021
  5. Cheng, C.H., F.F. Yang, R.Z. Ling, S.A. Liao, Y.T. Miao, C.X. Ye and A.L. Wang. 2015. Effects of ammonia exposure on apoptosis, oxidative stress and immune response in pufferfish (Takifugu obscurus). Aquatic Toxicology 164: 61-71. https://doi.org/10.1016/j.aquatox.2015.04.004
  6. Devos, A., L.J. Dallas, C. Voiseux, C. Lecomte-pradines, A.N. Jha and B. Fievet. 2015. Assessment of growth, genotoxic responses and expression of stress related genes in the Pacific oyster Crassostrea gigas following chronic exposure to ionizing radiation. Marine Pollution Bulletin 95: 688-698. https://doi.org/10.1016/j.marpolbul.2015.03.039
  7. Feder, M.E. and G.E. Hofmann. 1999. Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annual Review of Physiology 61: 243-282. https://doi.org/10.1146/annurev.physiol.61.1.243
  8. Husmann, G., D. Abele, D. Monien, P. Monien, M. Kriews and E.E.R. Philipp. 2012. The influence of sedimentation on metal accumulation and cellular oxidative stress markers in the Antarctic bivalve Laternula elliptica. Estuarine, Coastal and Shelf Science 111: 48-59. https://doi.org/10.1016/j.ecss.2012.06.003
  9. Jeon, M.A., J.C. Kang and J.S. Lee. 2013. Concentration of heavy metal and alteration of reproductive and histological biomarker of Mytilus galloprovincialis in Gamak bay of the southern coast of Korea. The Korean Journal of Malacology 29: 33-41. https://doi.org/10.9710/kjm.2013.29.1.33
  10. Ji, C., L. Cao and F. Li. 2015. Toxicological evaluation of two pedigrees of clam Ruditapes philippinarum as bioindicators of heavy metal contaminants using metabolomics. European Jornal of Pharmacology-Environmental Toxicology and Pharmacology Section 39: 545-554.
  11. Kalmar, B. and L. Greensmith. 2009. Induction of heat shock proteins for protection against oxidative stress. Advanced Drug Delivery Reviews 61: 310-318. https://doi.org/10.1016/j.addr.2009.02.003
  12. Kim, B.K., M.O. Lee and S.J. Park. 2012. Characteristics of water temperature and salinity variations, and seawater exchange in Gamak bay. JKOSME 15: 101-110. https://doi.org/10.7846/JKOSMEE.2012.15.2.101
  13. Kim, C.W., E.O. Kim, H.D. Jeong, C.G. Jung, M.W. Park and S.G. Son. 2009. Variation of body composition and survival rate according to spawning of pacific oyster, (Crassostrea gigas) in Gamak bay. Korean Journal of Fisheries and Aquatic Science 42: 481-486. https://doi.org/10.5657/kfas.2009.42.5.481
  14. Kim, W.S., K. Park, C. Nikapitiya and I.S. Kwak. 2014. Ecotoxicological end-points on intertidal mud crab, Macrophthalmus japonicus, following PFOS (Perfluorooctane sulfonate) exposure. Korean Journal of Environmental Biology 32: 311-318. https://doi.org/10.11626/KJEB.2014.32.4.311
  15. Kim, W.S., R. Kim, C. Nikapitiya, K. Park and I.S. Kwak. 2015. The molecular biomarker genes expression of rearing species Chironomus riparious and field species Chironomus plumosus exposure to heavy metals. Korean Journal of Ecology and Environment 48: 86-94. https://doi.org/10.11614/KSL.2015.48.2.086
  16. Kim, Y. and E.N. Powell. 2009. Effects of climate variability on interannual variation in parasites, pathologies, and physiological attributes of bivalves from the U.S. East, Gulf, and West Coasts. Environmental Bioindicators 4: 67-96. https://doi.org/10.1080/15555270802708830
  17. Knowles, G., J. Handlinger, B. Jones and N. Moltschaniwskyj. 2014. Hemolymph chemistry and histopathological changes in Pacific oyster (Crassostresa gigas) in response to low salinity stress. Journal of Invertebrate Pathology 121: 78-84. https://doi.org/10.1016/j.jip.2014.06.013
  18. Labreuche, Y., C. Lambert, P. Soudant, V. Boulo, A. Huvet and J.L. Nicolas. 2006. Cellular and molecular hemocyte responses of the pacific oyster, Crassostrea gigas, following bacterial infection with vibrio aestuanianus strain 01/32. Microbes and Infection 8: 2715-2724. https://doi.org/10.1016/j.micinf.2006.07.020
  19. Lee, K.H. and K.D. Cho. 1990. Distribution of the temperature and salinity in Kamak Bay. Korean Journal of Fisheries and Aquatic Sciences 23: 25-39.
  20. Lee, M.O., B.K. Kim, S.J. Park and J.K. Kim. 2005. Marine environment of Gamak Bay by field observations and numerical experiments. Journal of the Korean Socety for Marine Environmental Engineerning 11: 109-114.
  21. Lezcano, A.H., M.L.R. Quiroga, A.L. Liberoff and S.V.D. Molen. 2015. Marine pollution effects on the southern surf crab Ovalipes trimaculatus (Crustacea: Brachyura: Polybiidae) in Patagonia Argentina. Marine Pollution Bulletin 91: 524-529. https://doi.org/10.1016/j.marpolbul.2014.09.038
  22. Li, S., Y. Liu, C. Liu, J. Huang, G. Zheng, L. Xie and R. Zhang. 2015. Morphology and classification of hemocytes in Pinctada fucata and their responses to ocean acidification and warming. Fish and Shellfish Immunology 45: 194-202. https://doi.org/10.1016/j.fsi.2015.04.006
  23. Limon-Pacheco, J. and M.E. Gonsebatt. 2009. The role of antioxidants and antioxidant-related enzymes in protective responses to environmentally induced oxidative stress. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 674: 137-147. https://doi.org/10.1016/j.mrgentox.2008.09.015
  24. Lu, R.C., M.S. Tan, H. Hwang, A.M. Xie, J.T. Yu and L. Tan. 2014. Heat shock protein 70 in Alzheimer's disease. Biomed Research International 435203.
  25. Luo, L., C. Ke, X. Guo, B. Shi and M. Huang. 2014. Metal accumulation and differentially expressed proteins in gill of oyster (Crassostrea hongkongensis) exposed to long-term heavy metal-contaminated estuary. Fish and Shellfish Immunology 38: 318-329. https://doi.org/10.1016/j.fsi.2014.03.029
  26. Mizrahi, T., S. Goldenberg, J. Heller and Z. Arad. 2015. Natural variation in resistance to desiccation and heat shock protein expression in the land snail Theba pisana along a climate gradient. Physiological and Biochemical Zoology 88: 66-80. https://doi.org/10.1086/679485
  27. Nikapitiya, C., W.S. Kim, K. Park and I.S. Kwak. 2014. Identification of potential markers and sensitive tissues for low or high salinity stress in an intertidal mud crab (Macrophthalmus japonicus). Fish and Shellfish Immunology 41: 407-416. https://doi.org/10.1016/j.fsi.2014.09.018
  28. Nikapitiya, C., W.S. Kim, K. Park, J. Kim, M.O. Lee and I.S. Kwak. 2015. Chitinase gene reponses and tissue sensitivity in an intertidal mud crab (Macrophthalmus japonicus) following low or high salinity stress. Cell Stress and Chaperones 20: 517-526. https://doi.org/10.1007/s12192-015-0576-1
  29. Park, K. and I.S. Kwak. 2013. Expression of stress response HSP70 gene in Asian paddle crabs, Charybdis japonica, exposure to endocrine disrupting chemicals, bisphenol-A (BPA) and 4-nonylpheonl (NP). Ocean Science Journal 48: 207-214. https://doi.org/10.1007/s12601-013-0017-y
  30. Park, K. and I.S. Kwak. 2014. The effect of temperature gradients on endocrine signaling and antioxidant gene expression during Chironomus riparious development. Science of the Total Environment 470-471: 1003-1011. https://doi.org/10.1016/j.scitotenv.2013.10.052
  31. Park, K., H.W. Bang, J. Park and I.S. Kwak. 2009. Ecotoxicological multilevel-evaluation of the effects of fenbendazole exposure to Chironomus riparious larvae. Chemosphere 77: 359-367. https://doi.org/10.1016/j.chemosphere.2009.07.019
  32. Park, K., R. Kim, J.J. Park, H.C. Shin, J.S. Lee, H.S. Cho, Y.G. Lee, J.K. Kim and I.S. Kwak. 2012. Ecotoxicological evaluation of tributyltin toxicity to the equilateral venus clam, Gomphina veneriformis. Fish and Shellfish Immunology 32: 426-433. https://doi.org/10.1016/j.fsi.2011.11.031
  33. Ricketts, C.D., W.R. Bates and S.D. Reid. 2015. The effects of acute waterborne exposure to sublethal concentrations of molybdenum on the stress response in rainbow trout, Oncorhynchus mykiss. Plos one 10: e0115334. https://doi.org/10.1371/journal.pone.0115334
  34. Schintu, M., L. Durante, A. Maccioni, P. Meloni, S. Degetto and A. Contu. 2008. Measurement of environmental tracemetal levels in Mediterranean coastal areas with transplanted mussels and DGT techniques. Marine Pollution Bulletin 57: 832-837. https://doi.org/10.1016/j.marpolbul.2008.02.038
  35. Shin, Y.K., J.H. Im, M.H. Son and E.O. Kim. 2012. Seasonal variation in biochemical composition and gonadal development of ark shell, Scapharca broughtonii form Gamag bay of Southern coast, Korea. The Korean Journal of Malacology 28: 73-79. https://doi.org/10.9710/kjm.2012.28.1.073
  36. Tamura, K., G. Stecher, D. Peterson, A. Filipski and S. Kumar. 2013. MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 30: 2725-2729. https://doi.org/10.1093/molbev/mst197
  37. Verlecar, X.N., K.B. Jena and G.B.N. Chainy. 2007. Biochemical markers of oxidative stress in perna biridis exposed to mercury and temperature. Chemico-Biological Interactions 167: 219-226. https://doi.org/10.1016/j.cbi.2007.01.018
  38. Wang, Q., L. Zhangm, L. Zhao, L. You and H. Wu. 2012. Two goose-type lysozymes in Mytilus galloprovincialis: possible function diversification and adaptive evolution. Plos one 7: e45148. https://doi.org/10.1371/journal.pone.0045148
  39. Wei, L., Q. Wang, H. Wu, G. Ji and J. Zhao. 2014. Proteomic and metabolomics responses of Pacific oyster Crassostrea gigas to elevated $pCO_2$ exposure. Journal of Proteomics 112: 83-94.
  40. Xu, D., L. Sun, S. Liu, L. Zhang and H. Yang. 2015. Histological ultrastructural and heat shock protein 70 (HSP70) responses to heat stress in the sea cucumber Apostichopus japonicus. Fish and Shellfish Immunology 45: 321-326. https://doi.org/10.1016/j.fsi.2015.04.015
  41. Yevich, P.P. and C.A. Yevich. 1994. Use of histopathology in biomonitoring marine bivalves. In: Kramer, K.J.M. (ed.), Biomonitoring of Coastal Waters and Estuaries. CRC. pp. 179-204.
  42. Yoon, H., Y.K. An and S. Choi. 2011. Species composition and seasonal variation of fish assemblages in Sargassum bed in Gamak bay, Korea. Journal of the Korean Society of Marine Environment & Safety 77: 15-21.
  43. Yoon, S.P., R.H. Jung, S.S. Kim, J.S. Lee, J.S. Park, W.C. Lee and W.J. Choi. 2007. Characteristics of benthic environment and polychaete communities of Gamak bay, Korea. The Sea 12: 287-304.
  44. Zhang, L., X. Liu, L. Chen, L. You, D. Pei, M. Cong, J. Zhao, G. Li, D. Liu, J. Yu and H. Wu. 2011. Transcriptional regulation of selenium-dependent glutathione peroxidase from Venerupis philippinarum in response to pathogen and contaminants challenge. Fish and Shellfish Immunology 31: 831-837. https://doi.org/10.1016/j.fsi.2011.07.026
  45. Zhang, Z. and Q. Zhang. 2012. Molecular cloning, characterization and expression of heat shock protein 70 gene from the oyster Crassostrea hongkongensis responding to thermal stress and exposure $Cu^{2+}$ and malachite green. Gene 497: 172-180. https://doi.org/10.1016/j.gene.2012.01.058