Episodes of summer mortality of the Kumamoto oyster Crassostrea sikamea are a major problem for its cultivation. Expression of the heat shock protein 70 (HSP70) is induced by various environmental stresses, including heat. We cloned and sequenced hsp70 complementary DNA from C. sikamea to investigate the relationship between hsp70 expression and heat tolerance in this oyster. Quantitative real-time polymerase chain reaction was performed using gill tissue dissected from oysters before and after heat shock for 1 h. The results showed hsp70 expression was faster and greater in oysters cultured at 20–22 °C than at 10–12 °C, and survival was lower among oysters cultured at 20–22 °C than at 10–12 °C. Moreover, heat tolerance was investigated by a 1-h pre-heat treatment, followed by exposure to heat shock conditions 5 days later. Survival was higher and hsp70 expression was notably lower in oysters that received the pre-heat treatment compared with those that did not. We conclude that a pre-heat treatment of only 1 h may be useful for inducing heat tolerance in C. sikamea, and that a low level of hsp70 expression after heat shock is an important index in selecting for high heat tolerance in these oysters.
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Srivastava P (2002) Roles of heat-shock proteins in innate and adaptive immunity. Nat Rev Immunol 2:185–194
Feder ME, Hofman GE (1999) Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annu Rev Physiol 61:243–282
Gething MJ, Sambrook J (1992) Protein folding in the cell. Nature 355:33–45
Piano A, Asirelli C, Caselli F, Fabbri E (2002) Hsp70 expression in thermally stressed Ostrea edulis, a commercially important oyster in Europe. Cell Stress Chap 7:250–257
Lindquest SL (1986) The heat-shock responses. Annu Rev Biochem 55:1151–1191
Sorensen JG, Kristensen TN, Loeschcke V (2003) The evolutionary and ecological role of heat shock proteins. Ecol Lett 6:1025–1037
Fabbri E, Valbonesi P, Franzellitti S (2008) HSP expression in bivalves. ISJ 5:135–161
Boutet I, Tanguy A, Rousseau S, Auffert M, Moraga M (2003) Molecular identification and expression of heat shock cognate 70 (hsc70) and heat shock protein (hsp70) genes in the Pacific oyster Crassotrea gigas. Cell Stress Chap 8:76–85
Boutet I, Tanguy A, Moraga D (2003) Organization and nucleotide sequence of the European flat oyster Ostrea edulis heat shock cognate 70 (hsc70) and heat shock protein (hsp70) genes. Aquat Toxicol 65:221–225
Clegg JS, Uhlingher KR, Jackson SA, Cherr GN, Rifkin E, Friedman CS (1998) Induced thermotolerance and heat shock protein-70 family in Pacific oyster Crassostrea gigas. Mol Mar Biol Biotechnol 7:21–30
Piano A, Franzellitti S, Tinti F, Fabbri E (2005) Sequencing and expression pattern of inducible heat shock gene products in the European flat oyster, Ostrea edulis. Gene 361:119–126
Patterson HK, Boettcher A, Carmichael RH (2014) Biomarkers of dissolved oxygen stress in oysters: a tool for restoration and management efforts. PLOS One 9:e104440
Shamseldin A, Clegg J, Friedman C, Cherr N, Pillai C (1997) Induced thermotolerance in the pacific oyster, Crassostrea gigas. J Shellfish Res 16:487–491
Hamdoun AM, Cheney DP, Cherr GN (2003) Phenotypic plasticity of HSP70 and HSP70 gene expression in the Pacific oyster (Crassostrea gigas): implications for thermal limits and induction of thermal tolerance. Biol Bull 205:160–169
Jackson SA, Uhlinger KR, Clegg JS (2011) Duration of induced thermal tolerance and tissue-specific expression of hsp/hsc70 in the eastern oyster, Crassostrea virginica and the Pacific oyster, Crassostrea gigas. Aquaculture 317:168–174
Encomio VG, Chu FLE (2007) Heat shock protein (hsp70) expression and thermal tolerance in sublethally heat-shocked eastern oysters Crassostrea virginica infected with the parasite Perkinsus marinus. Dis Aquat Org 76:251–260
Usuki H (2002) Evaluation of characteristics and preservation of Pacific oyster, Crassostrea gigas, in view of the genetic resources. Bull Fish Res Agency 4:40–104
Imai T, Sakai S (1961) Study if breeding the Japanese oyster, Crassotrea gigas. Tohoku J Agric Res 12:125–171
Numachi K (1971) Biological research on the oyster. In: Imai T (ed) Aquaculture in shallow seas: progress in shallow sea culture. Koseisha Koseikaku, Tokyo, pp 82–105
Banks MA (1993) Discrimination between closely related Pacific oyster species (Crassostrea) via mitochondrial DNA sequences coding for large subunit rRNA. Mol Mar Bio Biotech 2:129–136
Beattie JH, Kenneth K, Chew K, William K, Harshberger K (1980) Differential survival of selected strains of Pacific oysters (Crassotrea gigas) during summer mortality. Proc Natl Shellfish Assoc 70:184–189
Degremont L, Bedier E, Soletchnik P, Ropert M, Huvet A, Moal J, Samin JF, Boudry P (2005) Relative importance of family, site, and field placement timing on survival, growth, and yield of hatchery-produced Pacific oyster spat (Crassostrea gigas). Aquaculture 249:213–229
Degremont L, Bedier E, Boudry P (2010) Summer mortality of hatchery-produced Pacific oyster spat (Crassostrea gigas). II. Response to selection for survival and its influence on growth and yield. Aquaculture 299:21–29
Martinetz J, Vasquez R, Renteria Y (2012) Early gametogenesis of Kumamoto oyster (Crassostrea sikamea). Hidrobiologica 2:181–184
Farcy E, Voiseux C, Lebel JM, Fiévet B (2009) Transcriptional expression levels of cell stress marker genes in the Pacific oyster Crassostrea gigas exposed to acute thermal stress. Cell Stress Chap 14:371–380
Zhang Z, Zhang Q (2012) Molecular cloning, characterization and expression of heat shock protein 70 gene from the oyster Crassotrea hongkongensis responding to thermal stress and exposure of Cu2+ and malachite green. Gene 497:172–180
De Decker S, Saulnier D (2011) Vibriosis induced by experimental cohabitation in Crassostrea gigas: evidence of early infection and down-expression of immune-related genes. Fish Shellfish Immunol 34:939–945
Gupta RS, Singh B (1994) Phylogenetic analysis of 70kD heat shock protein sequences suggests a chimeric origin for the eukaryotic cell nucleus. Curr Biol 4:1104–1114
Laursen JR, Liu H, Wu XJ, Yoshino TP (1997) Heat-shock response in a molluscan cell line: characterization of the response and cloning of an inducible HSP70 cDNA. J Invertebr Pathol 70:226–233
Samain JF, Degremont L, Soletchinik P, Haure J, Bedier E, Ropert M, Moal J, Huvet A, Bacca H, Wormhoudt V, Delaporte M, Costil K, Pouvreau S, Lambert C, Soudant P, Nicolas JL, Roux FL, Renault T, Gagnaire B, Geret F, Boutet I, Burgeot T, Bourdry P (2007) Genetically based resistance to summer mortality in the Pacific oyster (Crassostrea gigas) and its relationship with physiological, immunological characteristics and infection processes. Aquaculture 268:227–243
Song HM, Mu XD, Gu DE, Luo D, Yang YX, Xu M, Luo JR, Zhang JE, Hu YC (2014) Molecular characteristics of the HSP70 gene and its differential expression in female and male golden apple snails (Pomacea canaliculata) under temperature stimulation. Cell Stress Chap 19:579–589
De Maio A (1999) Heat shock proteins: facts, thoughts, and dreams. Shock 11:1–12
We thank the staff of the Kumamoto Prefectural Fisheries Research Center for the rearing of C. sikamea.
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Nagata, T., Sameshima, M., Uchikawa, T. et al. Molecular cloning and expression of the heat shock protein 70 gene in the Kumamoto oyster Crassostrea sikamea . Fish Sci 83, 273–281 (2017). https://doi.org/10.1007/s12562-017-1064-6
- Stress response
- Quantitative real-time polymerase chain reaction
- Heat tolerance
- Pre-heat treatment
- Molecular chaperone