Foraminifera is the most important temperature proxy of the ocean on long time scales. However, the absence of temperature-controlled experiments at different water depths hinders the advancement of paleotemperature reconstruction with foraminifera from the continental shelf. For the first time, this study investigated the response of benthic foraminifera to temperature change using microcosm culture and metabarcoding. Foraminiferal communities from three continental stations at varying water depths (6.0, 9.2, and 26.0 m) were cultured under five temperature gradients (6, 12, 18, 24, and 30°C), with each treatment performed in triplicate. The foraminifera were fed with microalgae every 4 days, and the filtered seawater (through 0.22 μm pores), acting as a medium, was changed accordingly. The experiment lasted for 80 days, and 47 DNA samples were obtained and analyzed, including three insitu samples. The results showed that foraminifera adjusted its growth rate within the low-temperature range and adopted an r-strategy to cope with high-temperature stress. In addition, the foraminifera from deeper water stations exhibited a pronounced vulnerability to diminishing read counts. The read counts, operational taxonomic units (OTU) counts and Margalef index of foraminifera and the read counts of Rotaliida exhibited a remarkably positive correlation with temperature. The recommended relationships were described as read counts = 1314.75*T + 44754.51; OTU counts = 1.13*T + 44.26; Margalef index =1.13*T + 44.26. This study established the first quantitative relationship between temperature and foraminifera molecular parameters that holds significant implications for long-time paleotemperature calibration in climate change.
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