Molecular quantification of copepod Acartia erythraea feeding on different algae preys

Simin Hu Tao Li Hui Huang Sheng Liu

Simin Hu, Tao Li, Hui Huang, Sheng Liu. Molecular quantification of copepod Acartia erythraea feeding on different algae preys[J]. Acta Oceanologica Sinica, 2023, 42(9): 125-131. doi: 10.1007/s13131-022-2115-0
Citation: Simin Hu, Tao Li, Hui Huang, Sheng Liu. Molecular quantification of copepod Acartia erythraea feeding on different algae preys[J]. Acta Oceanologica Sinica, 2023, 42(9): 125-131. doi: 10.1007/s13131-022-2115-0

doi: 10.1007/s13131-022-2115-0

Molecular quantification of copepod Acartia erythraea feeding on different algae preys

Funds: The National Natural Science Foundation of China under contract Nos 41806188 and 42176118; the Science and Technology Planning Project of Guangdong Province, China under contract No. 2020B1212060058; the Key Special Project for Introduced Talents Team of Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou) under contract No. GML2019ZD0404.
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  • Figure  1.  Extraction and linearization of plasmids inserted target 18S rDNA fragment (A), validation of the specificy primers in RT-PCR assay with plasmids above (B) and extracted DNA from fixed copepod (C) as template. pT, lT, pA, lA, pP, lP in A and B represent the templates from plasmid, linear plasmid inserted with 18S rDNA fragment of Thalassiosira weissflogii, Alexandrium catenella, and Prorocentrum shikokuense, respectively. 10× and 5× indicate the dilution time of template. a, b, and c in C display the PCR product amplified with species-specific primers of PRD, ALC, and THW, respectively. M: DNA marker 2000. N: negative. DDW: double distilled water; PRD: P. shikokuense; THW: T. weissflogii; ALC: A. catenella; Dun: Dunaliella sp. (as the control group); pT, lT: plasmid and linearization of 18S rDNA of T. weissflogii; pA, lA: plasmid and linearization of 18S rDNA of A. catenella. pP, lP: plasmid and linearization of 18S rDNA of P. shikokuense.

    Figure  2.  Quantification of microalgae 18S rDNA gene copies in fixed algae (algal cells left after feeding) and copepods based on the standards of three microalgae: Thalassiosira weissflogii (THW) (a), Prorocentrum shikokuense (PRD) (b), and Alexandrium catenella (ALC) (c). Linear plasmids inserted with target 18S rDNA fragment are used as templates to make quantitative standards for estimating cells remaining after the feeding experiment and in the copepod.

    Figure  3.  Ingestion rate (IR), digestion rate (DR), and assimilation of Acartia erythraea on different algal species based on qPCR results. THW: Thalassiosira weissflogii; PRD: Prorocentrum shikokuense; ALC: Alexandrium catenella.

    Table  1.   Species-specific primers used in this study

    Algae species Primer forward (5'–3') Primer reverse (5'–3') Amplicon size/bp GenBank Acc. No.
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    Table  2.   Ingestion rate of Acartia erythraea based on cell counting

    Algae species Cell density before/
    Cell density after/(cells·mL−1) Copepod individuals Incubation time/
    Ingestion rate/
    Thalassiosira weissflogii 1.45 (± 0.09)×104 1.04 (± 0.07)×104 30 15 4.31 (± 1.61)×103
    Prorocentrum shikokuense 1.05 (± 0.07)×103 0.88 (± 0.11)×102 30 10 1.53 (± 0.13)×103
    Alexandrium catenella 2.08 (± 0.13)×103 1.37 (± 0.11)×103 30 8 1.43 (± 0.04)×103
    Note: before and after the feeding experiment.
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  • 收稿日期:  2022-05-18
  • 录用日期:  2022-08-23
  • 网络出版日期:  2023-09-28
  • 刊出日期:  2023-09-01