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Development and Testing of Species-Specific Primers for Detecting the Presence of the Northern Pacific Sea Star (Asterias amurensis) from Environmental DNA

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Abstract

The starfish Asterias amurensis, a well-known predator of molluscan species in intertidal ecosystems, has caused substantial ecological and economic losses in North China such as offshore Qingdao. Effective monitoring and prevention measures are urged to minimize its negative impacts. Compared with traditional biomonitoring methods, environmental DNA technology has emerged as a powerful and cost-efficient tool for inferring species’ presence and abundance. In this study, we developed a pair of species-specific primers (i.e., Ast-F and Ast-R) for the A. amurensis mitochondrial COI gene and tested its utility in amplifying and quantifying the DNA fragments from environmental samples under both laboratory and field conditions. The results of controlled water tank experiments demonstrated that the amount of eDNA released by A. amurensis was positively related to its biomass; after the removal of the starfish, the eDNA degraded significantly in 24 h and remained detectable for 8 days. The number of eDNA copies enriched tended to increase with smaller pore size of filter membrane and larger volume of filtered water. For field tests, we confirmed the validation of our approach in six locations in Qingdao by filtering 1000 ml water per sample with a 0.45-µm pore size filtration. All the amplification products generated a single and bright band via gel electrophoresis, and the quantitative PCR results unveiled significant differences in eDNA copies. This study provided an eDNA-based approach for investigating the distribution and biomass of A. amurensis, which may help to formulate early warning and management strategies in coastal Qingdao and other regions.

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Acknowledgements

We thank Chengbin Liu and Zhichao Huang for the sample collection.

Funding

This work was funded by grants from the National Key Research and Development Program, China (2022YFC3106301), and the Young Talent Program of Ocean University of China (No. 862201013143).

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Chenhu Yang processed samples, performed statistical analysis, and wrote the manuscript. Yanzhen Du designed the primers and prepared experiments. Gang Ni and Xiaoqi Zeng initiated the study, obtained funding and conducted the revision. All authors approved the final version.

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Correspondence to Gang Ni.

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Yang, C., Du, Y., Zeng, X. et al. Development and Testing of Species-Specific Primers for Detecting the Presence of the Northern Pacific Sea Star (Asterias amurensis) from Environmental DNA. Mar Biotechnol 26, 215–222 (2024). https://doi.org/10.1007/s10126-024-10292-1

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