An internal transcribed spacer 2 gene fragment, as a suitable phylogenetic marker in Hyalomma and Rhipicephalus (Acari: Rhipicephalinae) species

Document Type : Research articles

Authors

• Zakkyeh Telmadarraiy ¹
• Mohammad Nasrabadi ¹
• Mohammad Mehdi Sedaghat ¹
• Asadollah Hosseini-Chegeni ²
• Saedeh Sadat Hashemi-Aghdam ³
• Reza Hosseini ⁴
• S. Gholami ¹

¹ Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

² Department of Plant Protection, Faculty of Agriculture, University of Lorestan, Lorestan, Iran

³ Deptartment of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran

⁴ Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran

10.22067/ijab.v12i2.49415

Abstract

Hyalomma and Rhipicephalus (Acari: Ixodidae: Rhipicephalinae) are the most important ectoparasites and biological vector of pathogen agents. The taxonomy of some Hyalomma and Rhipicephalus species is more debatable. The traditional taxonomy of ticks have been revolutionized using polymerase chain reaction. The specificity, efficiency and accuracy of PCR is highly dependent on the oligonucleotide primers. In this study, five primers designed for amplification of a fragment ITS2 so-called TAH-1, TAH-2, TRH (for Hyalomma) and TAH-3, TAH-4 (for Rhipicephalus). Successfully, 24 and 29 ITS2 sample was sequenced using these primers. As well as, an ITS2 phylogenetic tree were constructed using BEAST basis on the Bayesian Inference (BI) method. The most Hyalomma and Rhipicephalus species well differentiated using ITS2 fragment since the designed primer fail to amplify gene target in few cases.

Keywords

• PCR
• Primer
• ITS2
• Hyalomma
• Rhipicephalus
• Phylogenetic relationship

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References

Barker, S.C. (1998) Distinguishing species and populations of Rhipicephaline ticks with ITS 2 ribosomal RNA. J. Parasitol., 84 (5): 887–892.

 

Barker, S.C. & Murrell, A. (2008). Systematics and evolution of ticks with a list of valid genus and species names. In: Bowman, A.S. and Nuttall, P.A. (Eds.), Ticks: biology, disease and control. Cambridge University Press, Cambridge UK, pp. 1–39.

 

Burpo, F.J. (2001) A critical review of PCR primer design algorithms and crosshybridization case study. Biochemistry, 218 1-12.

 

Chitimia, L., Lin, R.Q., Cosoroaba, I., Braila, P., Song, H.Q. & Zhu, X.Q. (2009) Molecular characterization of hard and soft ticks from Romania by sequences of the internal transcribed spacers of ribosomal DNA. Parasitology Research, 105 907-911.

 

Dantas-Torres, F., Latrofa, M.S., Annoscia, G., Giannelli, A., Parisi, A. & Otranto, D. (2013) Morphological and genetic diversity of Rhipicephalus sanguineus sensu lato from the new and old worlds. Parasit. Vectors, 6 (6): 1-17.

 

Dergousoff, S.J. & Chilton, N.B. (2007) Differentiation of three species of Ixodid tick, Dermacentor andersoni, D. variabilis and D. albipictus, by PCR-based approaches using markers in ribosomal DNA. Mol Cell Probes, 21 (5): 343–348.

 

Drummond, A.J., Suchard, M.A., Xie, D. & Rambaut, A. (2012) Bayesian phylogenetics with BEAUti and the BEAST 1.7. Molecular Biology and Evolution, 29 (8): 1969-1973.

 

Fukunaga, M., Yabuki, M., Hamase, A., Oliver, J.J.H. & Nakao, M. (2000 ) Molecular phylogenetic analysis of Ixodid ticks based on the ribosomal DNA spacer, internal transcribed spacer 2, sequences. Journal of Parasitology, 86 (1): 38–43.

 

Ganjali, M., Haddadzadeh, H.R. & Shayan, P. (2011) Nucleotide sequence analysis of the second internal transcribed spacer (ITS2) in Hyalomma anatolicum anatolicum in Iran. International Journal of Veterinary Research, 5 (2): 89-93.

 

Hillis, D.M. & Dixon, M.T. (1991) Ribosomal DNA: molecular evolution and phylogenetic inference. The quarterly review of biology, 66 (4): 411-453.

 

Hlinka, O., Murrell, A. & Barker, S.C. (2002) Evolution of the secondary structure of the rRNA internal transcribed spacer 2 (ITS2) in hard ticks (Ixodidae, Arthropoda). Heredity, 88 275-279.

 

Hosseini-Chegeni, A., Hosseine, R., Tavakoli, M., Telmadarraiy, Z. & Abdigoudarzi, M. (2013) The Iranian Hyalomma (Acari: Ixodidae) with a key to the identification of male species. Persian J. Acarol., 2 (3): 503–529.

 

Labruna, M.B., Marrelli, M.T., Heinemann, M.B., Fava, A.B., Cortez, A., Soares, R.M., Sakamoto, S.M., Richtzenhain, L.J., Marinotti, O. & Schumaker, T.T.S. (2002) Taxonomic Status of Ixodes didelphidis (Acari: Ixodidae). J Med Entomol, 39 (1): 135–142.

 

Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A. & Lopez, R. (2007) Clustal W and Clustal X version 2.0. Bioinformatics, 23 (21): 2947-2948.

 

Lv, J., Wu, S., Zhang, Y., Chen, Y., Feng, C., Yuan, X., Jia, G., Deng, J., Wang, C., Mei, L. & Lin, X. (2014) Assessment of four DNA fragments (COI, 16S rDNA, ITS2, 12S rDNA) for species identification of the Ixodida (Acari: Ixodida). Parasite Vector, 7 (1): 1–12.

 

McLain, D.K., Wesson, D.M., Collins, F.H. & Oliver Jr, J.H. (1995) Evolution of the rDNA spacer, ITS 2, in the ticks Ixodes scapularis and I. pacificus (Acari: Ixodidae). Heredity, 75 (3): 303-319.

 

Mcpherson, M. & Moller, S. (2006) PCR. Taylor & Francis, New York USA. pp. 292.

 

Moshaverinia, A., Shayan, P., Nabian, S. & Rahbari, S. (2009) Genetic evidence for conspecificity between Dermacentor marginatus and Dermacentor niveus. Parasitology Research, 105 1125–1132.

 

Murrell, A., Campbell, N.J.H. & Barker, S. (2001) Recurrent gains and losses of large (84–109 bp) repeats in the rDNA internal transcribed spacer 2 (ITS2) of rhipicephaline ticks. Insect Molecular Biology, 10 (6): 587-596.

 

Navajas, M. & Fenton, B. (2000) The application of molecular markers in the study of diversity in acarology: a review. Experimental and Applied Acarology, 24 751-774.

 

Nicholas, K.B., Nicholas, H.B.J. & Deerfield, D.W. (1997) GeneDoc: analysis and visualization of genetic variation. Embnew News, 4 (1): 14.

 

Rees, D.J., Dioli, M. & Kirkendalla, L.R. (2003) Molecules and morphology: evidence for cryptic hybridization in African Hyalomma (Acari: Ixodidae). Mol Phyl Evol, 27 131–142.

 

Rich, S.M., Rosenthal, B.M., Telford, I., S. R., Spielman, A., Hartl, D.L. & Ayala, F.J. (1997) Heterogeneity of the internal transcribed spacer (ITS-2) region within individual deer ticks. Insect Molecular Biology, 6 (2): 123-129.

 

Sambrook, J. & Russell, D.W. (2001) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory Press, New York USA. pp. 737.

 

Sonenshine, D.E. (1993) Biology of ticks. Oxford University press, Oxford UK. pp. 465.

 

Song, S., Shao, R., Atwell, R., Barker, S. & Vankan, D. (2011) Phylogenetic and phylogeographic relationships in Ixodes holocyclus and Ixodes cornuatus (Acari: Ixodidae) inferred from COX1 and ITS2 sequences. International journal for parasitology, 41 (8): 871-880.

 

Tian, Z., Liu, G., Xie, J., Yin, H., Luo, J., Zhang, L., Zhang, P. & Luo, J. (2011) Discrimination between Haemaphysalis longicornis and H. qinghaiensis based on the partial 16S rDNA and the second internal transcribed spacer (ITS-2). Experimental and Applied Acarology, 54 (2): 165-172.

 

Yuryev, A. (2007) PCR primer design. Humana Press, New Jersey USA. pp. 432.

 

Zahler, M., Filippova, N.A., Morel, P.C., Gothe, R. & Rinder, H. (1997) Relationships between species of the Rhipicephalus sanguineus group: a molecular approach. J. Parasitol., 83 (2): 302-306.