Abstract
There has been no widely accepted DNA barcode for species identification of Demodex. In this study, we attempted to solve this issue. First, mitochondrial cox1-5′ and 12S gene fragments of Demodex folloculorum, D. brevis, D. canis, and D. caprae were amplified, cloned, and sequenced for the first time; intra/interspecific divergences were computed and phylogenetic trees were reconstructed. Then, divergence frequency distribution plots of those two gene fragments were drawn together with mtDNA cox1-middle region and 16S obtained in previous studies. Finally, their identification efficiency was evaluated by comparing barcoding gap. Results indicated that 12S had the higher identification efficiency. Specifically, for cox1-5′ region of the four Demodex species, intraspecific divergences were less than 2.0%, and interspecific divergences were 21.1–31.0%; for 12S, intraspecific divergences were less than 1.4%, and interspecific divergences were 20.8–26.9%. The phylogenetic trees demonstrated that the four Demodex species clustered separately, and divergence frequency distribution plot showed that the largest intraspecific divergence of 12S (1.4%) was less than cox1-5′ region (2.0%), cox1-middle region (3.1%), and 16S (2.8%). The barcoding gap of 12S was 19.4%, larger than cox1-5′ region (19.1%), cox1-middle region (11.3%), and 16S (13.0%); the interspecific divergence span of 12S was 6.2%, smaller than cox1-5′ region (10.0%), cox1-middle region (14.1%), and 16S (11.4%). Moreover, 12S has a moderate length (517 bp) for sequencing at once. Therefore, we proposed mtDNA 12S was more suitable than cox1 and 16S to be a DNA barcode for classification and identification of Demodex at lower category level.
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (Nos. 81471972 and 81271856).
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Supplementary material 1
Adults morphology of the four Demodex species (×400) (JPEG 60 kb)
Supplementary material 2
Sequence alignments of mtDNA cox1–5’region for the four Demodex species (PDF 104 kb)
Supplementary material 3
Sequence alignments of mtDNA 12S for the four Demodex species (PDF 27 kb)
Supplementary material 4
Blast results of mtDNA gene fragments for the four Demodex species (DOCX 42 kb)
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Hu, L., Yang, Y., Zhao, Y. et al. DNA barcoding for molecular identification of Demodex based on mitochondrial genes. Parasitol Res 116, 3285–3290 (2017). https://doi.org/10.1007/s00436-017-5641-5
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DOI: https://doi.org/10.1007/s00436-017-5641-5