Quantitative analysis of dinoflagellates and diatoms community via Miseq sequencing of actin gene and v9 region of 18S rDNA

Sci Rep. 2016 Oct 10:6:34709. doi: 10.1038/srep34709.

Abstract

Miseq sequencing and data analysis for the actin gene and v9 region of 18S rDNA of 7 simulated samples consisting of different mixture of dinoflagellates and diatoms were carried out. Not all the species were detectable in all the 18S v9 samples, and sequence percent in all the v9 samples were not consistent with the corresponding cell percent which may suggest that 18S rDNA copy number in different cells of these species differed greatly which result in the large deviation of the amplification. And 18S rDNA amplification of the microalgae was prone to be contaminated by fungus. The amplification of actin gene all was from the dinoflagellates because of its targeted degenerate primers. All the actin sequences of dinoflagellates were detected in the act samples except act4, and sequence percentage of the dinoflagellates in the act samples was not completely consistent with the dinoflagellates percentage of cell samples, but with certain amplification deviations. Indexes of alpha diversity of actin gene sequencing may be better reflection of community structure, and beta diversity analysis could cluster the dinoflagellates samples with identical or similar composition together and was distinguishable with blooming simulating samples at the generic level. Hence, actin gene was more proper than rDNA as the molecular marker for the community analysis of the dinoflagellates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics*
  • DNA, Protozoan / genetics
  • DNA, Ribosomal / genetics
  • Diatoms / classification*
  • Diatoms / genetics
  • Dinoflagellida / classification*
  • Dinoflagellida / genetics
  • High-Throughput Nucleotide Sequencing / methods*
  • Phylogeny
  • RNA, Ribosomal, 18S / genetics*
  • Sequence Analysis, DNA / methods*
  • Species Specificity

Substances

  • Actins
  • DNA, Protozoan
  • DNA, Ribosomal
  • RNA, Ribosomal, 18S