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Sequencing for complete rDNA sequences (18S, ITS1, 5.8S, ITS2, and 28S rDNA) of Demodex and phylogenetic analysis of Acari based on 18S and 28S rDNA

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Abstract

Due to the difficulty of DNA extraction for Demodex, few studies dealt with the identification and the phyletic evolution of Demodex at molecular level. In this study, we amplified, sequenced, and analyzed a complete (Demodex folliculorum) and an almost complete (D12 missing) (Demodex brevis) ribosomal DNA (rDNA) sequence and also analyzed the primary sequences of divergent domains in small-subunit ribosomal RNA (rRNA) of 51 species and in large-subunit rRNA of 43 species from four superfamilies in Acari (Cheyletoidea, Tetranychoidea, Analgoidea, and Ixodoidea). The results revealed that 18S rDNA sequence was relatively conserved in rDNA-coding regions and was not evolving as rapidly as 28S rDNA sequence. The evolutionary rates of transcribed spacer regions were much higher than those of the coding regions. The maximum parsimony trees of 18S and 28S rDNA appeared to be almost identical, consistent with their morphological classification. Based on the fact that the resolution capability of sequence length and the divergence of the 13 segments (D1–D6, D7a, D7b, and D8–D12) of 28S rDNA were stronger than that of the nine variable regions (V1–V9) of 18S rDNA, we were able to identify Demodex (Cheyletoidea) by the indels occurring in D2, D6, and D8.

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Author information

Correspondence to Ya-E Zhao.

Electronic supplementary material

Supplementary Material 1

The alignment of a complete nuclear rDNA sequence of Demodex folliculorum (18S, ITS-1, 5.8S, ITS-2 and 28S) and an almost complete sequence of Demodex brevis. ITS-1 and ITS-2 are noted by dark shades, and the 9 variable regions of SSU rDNA and 13 expansion segments of LSU rDNA are noted by light shades. (JPEG 2.30 mb)

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Zhao, Y., Wu, L., Hu, L. et al. Sequencing for complete rDNA sequences (18S, ITS1, 5.8S, ITS2, and 28S rDNA) of Demodex and phylogenetic analysis of Acari based on 18S and 28S rDNA. Parasitol Res 111, 2109–2114 (2012). https://doi.org/10.1007/s00436-012-3058-8

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Keywords

  • rDNA Sequence
  • Maximum Parsimony Tree
  • Transcribe Space Region
  • Meibomian Gland
  • Expansion Segment