Advanced approach to analyzing calcareous protists for present and past pelagic ecology: Comprehensive analysis of 3D-morphology, stable isotopes, and genes of planktic foraminifers

PLoS One. 2019 Mar 7;14(3):e0213282. doi: 10.1371/journal.pone.0213282. eCollection 2019.

Abstract

Marine protists play an important role in oceanic ecosystems and biogeochemical cycles. However, the difficulties in culturing pelagic protists indicate that their ecology and behavior remain poorly understood; phylogeographic studies based on single-cell genetic analyses have often shown that they are highly divergent at the biological species level, with variable geographic distributions. This indicates that their ecology could be complex. On the other hand, the biomineral (calcareous) shells of planktic foraminifers are widely used in geochemical analyses to estimate marine paleoenvironmental characteristics (i.e., temperature), because the shell chemical composition reflects ambient seawater conditions. Among the pelagic protists, planktic foraminifers are ideal study candidates to develop a combined approach of genetic, morphological, and geochemical methods, thus reflecting environmental and ecological characteristics. The present study precisely tested whether the DNA extraction process physically and chemically affects the shells of the planktic foraminifer Globigerinoides ruber. We used a nondestructive method for analyzing physical changes (micro-focus X-ray computed tomography (MXCT) scanning) to compare specimens at the pre- and post-DNA extraction stages. Our results demonstrate that DNA extraction has no significant effect on shell density and thickness. We measured stable carbon and oxygen isotopes on the shell of each individual in a negative control or one of two DNA-extracted groups and detected no significant differences in isotopic values among the three groups. Moreover, we evaluated isotopic variations at the biological species level with regard to their ecological characteristics such as depth habitat, life stages, and symbionts. Thus, our examination of the physiochemical effects on biomineral shells through DNA extraction shows that morphological and isotopic analyses of foraminifers can be combined with genetic analysis. These analytical methods are applicable to other shell-forming protists and microorganisms. In this study, we developed a powerful analytical tool for use in ecological and environmental studies of modern and past oceans.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animal Shells / anatomy & histology*
  • Animal Shells / metabolism*
  • Animals
  • Carbon Isotopes / analysis*
  • DNA, Protozoan / genetics
  • Ecology*
  • Foraminifera / chemistry
  • Foraminifera / genetics*
  • Foraminifera / metabolism
  • Oxygen Isotopes / analysis*
  • Phylogeography
  • Protozoan Proteins / genetics*

Substances

  • Carbon Isotopes
  • DNA, Protozoan
  • Oxygen Isotopes
  • Protozoan Proteins

Grants and funding

This work was supported by: YU: Japan Society for Promotion of Science KAKENHI 26400501; Education, Culture, Sports, Science and Technology Ministry of Japan special project costs “Four-Dimensional Kuroshio Marine Science (4D-KMS)”; TI: Japan Society for Promotion of Science KAKENHI 16H02944; KK: Japan Society for Promotion of Science 16H04961 and 15H05712.