Using quantitative PCR to determine the distribution of a semicryptic benthic diatom, Navicula phyllepta (Bacillariophyceae)

  • Créach V
  • Ernst A
  • Sabbe K
 et al. 
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Abstract

Evidence has accumulated during the last decade showing that many established diatom morpho-spe- cies actually consist of several semicryptic or truly cryptic species. As these species are difficult or even impossible to differentiate by microscopic analysis, there is virtually no information on how they behave in natural environments. In this study, we devel- oped a quantitative real-time PCR (qPCR) assay using TaqMan probess targeted to the internal tran- scribed spacer 1 (ITS1) to assess the spatial distri- bution and seasonal dynamics of an important component of the microphytobenthos of intertidal sediments. Navicula phyllepta Ku ¨tzing is a brackish- marine morpho-species with a cosmopolitan distri- bution. Axenic clones of this species were isolated from natural assemblages of benthic diatoms at dif- ferent intertidal stations in the Westerschelde estu- ary (The Netherlands). At least two distinct semicryptic species of N. phyllepta were present, as shown by differences in the quantity of DNA per cell, the ITS1 sequences and the copy number of ITS per cell. DNA and chl a concentrations extract- ed from sediment surface samples were closely cor- related, showing that the DNA used for subsequent analysis mostly belonged to the microalgal commu- nity. The results of real-time qPCR from sites throughout the estuary and over several seasons agreed well with microscopic counts. Additionally, the seasonal pattern of the two forms of N. phyllepta showed an overlapping, but unique distribution along

Author-supplied keywords

  • Diatoms
  • Estuary
  • Navicula phyllepta
  • Niche differentiation
  • Real-time qPCR
  • Semicryptic species

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Authors

  • Véronique Créach

  • Anneliese Ernst

  • Koen Sabbe

  • Bart Vanelslander

  • Wim Vyverman

  • Lucas J. Stal

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