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Expression Characterization of Stress Genes Under High and Low Temperature Stresses in the Pacific Oyster, Crassostrea gigas

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Abstract

As a characteristic sessile inhabitant of the intertidal zone, the Pacific oyster Crassostrea gigas occupies one of the most physically stressful environments on earth. With high exposure to terrestrial conditions, oysters must tolerate broad fluctuations in temperature range. However, oysters’ cellular and molecular responses to temperature stresses have not been fully characterized. Here, we analyzed oyster transcriptome data under high and low temperatures. We also identified over 30 key temperature stress-responsive candidate genes, which encoded stress proteins such as heat shock proteins and apoptosis-associated proteins. The expression characterization of these genes under short-term cold and hot environments (5 and 35 °C) and long-term cold environments (5 °C) was detected by quantitative real-time PCR. Most of these genes reached expression peaks during the recovery stage after 24 h of heat stress, and these genes were greatly induced around day 3 in long-term cold stress while responded little to short-term cold stress. In addition, in the second heat stress after 2 days of recovery, oysters showed milder expression in these genes and a lower mortality rate, which indicated the existence of plasticity in the oyster’s response to heat stress. We confirmed that homeostatic flexibility and anti-apoptosis might be crucial centers of temperature stress responses in oysters. Furthermore, we analyzed stress gene families in 11 different species and found that the linage-specific expansion of stress genes might be implicated in adaptive evolution. These results indicated that both plasticity and evolution played an important role in the stress response adaptation of oysters.

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Acknowledgments

We would like to thank Jinpeng Wang, Yingxiang Li, Baoyu Huang, and Wen Huang for sample collection and RNA preparation. We are grateful to Jun Liu and Dr. Tao Qu for their help in the primer design. We thank Dr. Haigang Qi and Dr. Fei Xu for their bioinformatic expertise. We appreciate the help from Mr. Bo Li and Mr. Xiaodong Fang of BGI-Shenzhen with the use of CAFE software.

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Authors and Affiliations

  1. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China

    Qihui Zhu, Linlin Zhang, Li Li, Huayong Que & Guofan Zhang

  2. University of Chinese Academy of Sciences, Beijing, China

    Qihui Zhu

  3. Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China

    Linlin Zhang, Huayong Que & Guofan Zhang

  4. National & Local Joint Engineering Laboratory of Ecological Mariculture, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China

    Qihui Zhu, Linlin Zhang, Li Li, Huayong Que & Guofan Zhang

  5. Laboratory for Marine Fisheries and Aquaculture, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China

    Li Li

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  1. Qihui Zhu
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Correspondence to Li Li or Guofan Zhang.

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Funding

This research was supported by National Natural Science Foundation of China (31572620), National Basic Research Program of China (973 Program) (2010CB126402), Earmarked fund for Modern Agro-industry Technology Research System (CARS-48) and Taishan Scholars Climbing Program of Shandong.

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Table S1

Shell morphological parameters of oyster samples (DOCX 238 kb)

Table S2

Housekeeping genes and their primer sequences used for endogenous control gene validation (XLSX 11 kb)

Table S3

GO terms and numbers of DEGs in Fig. 3 (DOCX 18 kb)

Table S4

The result of the program CAFE (XLSX 17 kb)

Table S5

Primers of temperature-responsive genes used in Q-RT-PCR (XLSX 12 kb)

Table S6

RPKM values of temperature responsive genes in the transcriptome data (DOCX 25 kb)

Table S7

Detailed Q-RT-PCR results for the responsive genes including individual values, mean values, standard deviations and statistical results (XLSX 15 kb)

Fig. S1

Average expression stability value (M) of housekeeping genes. (XLSX 130 kb)

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Zhu, Q., Zhang, L., Li, L. et al. Expression Characterization of Stress Genes Under High and Low Temperature Stresses in the Pacific Oyster, Crassostrea gigas . Mar Biotechnol 18, 176–188 (2016). https://doi.org/10.1007/s10126-015-9678-0

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