Genetic Characterization of Nematodirella cameli Based on 18S rDNA and Cytochrome c Oxidase Subunit 1 (CO1)

Abstract

To determine the phylogenic position and genetic diversity of Nematodirella cameli two portions of nuclear ribosomal DNA, 18S rDNA and mitochondrial DNA gene, the subunit 1 of cytochrome C oxidase gene (CO1) were sequenced and compared with those previously reported for other nematodes in Trichostrongylina. The phylogenetic trees constructed based upon the 18S rDNA sequences, yielded strong support for close relationship between the N. cameli and Nematodirus battus, with a high bootstrap value of 100%. In the present research, the level of sequence polymorphism among N. cameli isolates was higher for CO1 with 32 polymorphic sites compared to 18S rDNA sequence. Accordingly, molecular assays based on CO1 mitochondrial marker, demonstrated the existence of at least 11 distinct haplotypes (accession nos. JX305966 to JX305976) with an intraspecific diversity of 3-7% in Iran. Whereas, all of N. cameli samples examined herein (n=11), had a unique 18S sequence (accession no. JX305977). In addition, N. cameli CO1 sequences found in this study showed maximum identities to Haemonchus (88%) and Ostertagia (87%) in BLAST analysis for existing Trichostrongylina sequences. Further information is necessary to infer interspecific and intraspecific phylogenetic relationships between genera and species in Trichostrongylina. This study describes for the first time the nuclear 18S rDNA and mitochondrial CO1 sequence data from Nematodirella cameli species.

Keywords

• Nematodirella cameli, CO1, 18S rDNA, Camel, Iran

Nematodirella cameli‘nin 18S rDNA ve Sitokrom c Oksidaz Subunit 1 (C01)’e Dayalı Genetik Karakterizasyonu

Abstract

Nematodirella cameli’nin filojenik pozisyonu ve genetik çeşitliliğini belirlemek amacıyla nüklear ribozomal DNA’nın iki kısmı olan 18S rDNA ile sitokrom C oksidaz geninin (C01) subünite 1 olan mitokondriyal DNA geni sıralandı ve Trichostrongylina’daki diğer nematodlar için daha önce bildirilmiş olanlarla karşılaştırıldı. 18S rDNA sekanslarına dayandırılarak oluşturulan filogenetik ağaçlar,  %100’lük yüksek bir önyükleme değeri ile N. cameli ve Nematodirus battus arasında yakın bir bağlantı olduğuna dair güçlü bir destek oluşturdu. Mevcut çalışmada, N.cameli izolatları arasındaki sekans polimorfizm düzeyi,  C01’de 32 polimorfik konum ile 18S rDNA sekansına oranla daha yüksekti. Bununla bağlantılı olarak, İran’da C01 mitokondriyel markerine dayandırılarak yapılan moleküler testler, % 3-7 arasındaki bir intraspesifik çeşitlilik ile en az 11 belirgin haplotip varlığını gösterdi (Katılım No JX305966 ile JX305976 arası). Öte yandan, burada incelenen tüm N.cameli örneklerinin (n=11) özgün bir 185 dizilişi mevcuttu (Katılım No: JX305977). Buna ilaveten, bu çalışmada bulunan N.cameli C01 dizilişleri mevcut Trichostrongylina dizilişleri için BLAST analizinde en çok Haemonchus (%88) ve Ostertagia (%87) yapısı gösterdi. Trichostrongylina‘daki cinsler ve türler arasındaki interspesifik ve intraspesifik filogenetik ilişkileri gösterebilmek için daha ileri bilgiler gerekmektedir. Bu çalışma ilk kez olarak Nematodirella cameli türlerindeki nüklear 18S rDNA ve mitokondriyal C01 diziliş verilerini tanımlamaktadır.

Keywords

• Nematodirella cameli, CO1, 18S rDNA, Deve, İran

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