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Defense responses of sulfur dioxygenase to sulfide stress in the razor clam Sinonovacula constricta

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Abstract

Background

Sulfide is a well-known toxicant widely distributed in the culture environment. As a representative burrowing benthic bivalve, the razor clam Sinonovacula constricta is highly sulfide tolerant. Mitochondrial sulfide oxidation is an important way for sulfide detoxification, where sulfur dioxygenase (SDO) is the second key enzyme.

Objective

To investigate the mechanism of sulfide tolerance in S. constricta, the molecular characterization of its SDO (designated as ScSDO) was studied.

Methods

The cDNA sequence of ScSDO was cloned by RACE technique. The response of ScSDO in gills and livers of S. constricta was investigated during sulfide exposure (50, 150, and 300 μM sulfide) for 0, 3, 6, 12, 24, 48, 72, and 96 h by qRT-PCR. Moreover, the temporal expression of ScSDO protein in S. constricta gills after exposure to 150 μM sulfide was detected by Western blot. The subcellular location of ScSDO was identified by TargetP 1.1 prediction and Western Blot analysis.

Results

The full-length cDNA of ScSDO was 2914 bp, encoding a protein of 304 amino acids. The deduced ScSDO protein was highly conserved, containing the signature HXHXDH motif of the metallo-β-lactamase superfamily and two metal-binding sites, of which metal-binding site I is known to be the catalytically active center. Subcellular localization confirmed that ScSDO was located only in the mitochondria. Responding to the sulfide exposure, distinct time-dependent increases in ScSDO expression were detected at both mRNA and protein levels. Moreover, the gills exhibited a higher ScSDO expression level than the livers.

Conclusions

All of our results suggest that ScSDO plays an important role in mitochondrial sulfide oxidation during sulfide stress, making S. constricta highly sulfide tolerant. In addition, as a respiratory tissue, the gills play a more critical role in sulfide detoxification.

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (2018YFD0901405-5), the National Natural Science Foundation of China (31702317), Modern Agro-industry Technology Research System (CARS-49), Zhejiang Major Program of Science and Technology (2016C02055-9) and Key Research and Development project of Zhejiang Province(2019C02054).

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Authors and Affiliations

  1. Zhejiang Key Laboratory of Aquatic Germplasm Resources, Zhejiang Wanli University, Ningbo, Zhejiang, 315100, People’s Republic of China

    Caifang Chen, Yaoyao Shen, Jing He, Zhihua Lin & Yinghui Dong

  2. Ningbo Academy of Oceanology and Fishery, Ningbo, 315012, People’s Republic of China

    Weiliang Shen

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  1. Caifang Chen
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  2. Yaoyao Shen
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  3. Weiliang Shen
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Correspondence to Zhihua Lin or Yinghui Dong.

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Chen, C., Shen, Y., Shen, W. et al. Defense responses of sulfur dioxygenase to sulfide stress in the razor clam Sinonovacula constricta. Genes Genom 43, 513–522 (2021). https://doi.org/10.1007/s13258-021-01077-0

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