Internal Transcribed sequence (ITS) of Halocafeteria seosinensis (Bicosoecids)

Abstract

The aim of the current study was to characterize and identify the smaller subunit (18S) and Internal Transcribed Spacer (ITS) regions, ITS1 and ITS2 (ITS1-5.8S-ITS2) of Halocafeteria seosinensis (Bicosoecida). Halocafeteria was isolated from the salterns of Morocco and maintained using standard techniques. The ITS1 and ITS2 are non-conserved regions which were amplified using universal primers, ITS1 and ITS4. The purified PCR products were directly sequenced by Sanger dideoxy sequencing technology and submitted to GenBank. For the construction of cladogram, closely related 18S rDNA sequences obtained from GenBank were multiple aligned using MEGA 6 software and the cladogram constructed by maximum likelihood method with 10,000 bootstrapping (BP) repetitions. The phylogenetic relationship among the other related stramenopiles were also studied. The molecular characterization by PCR of 18S genes was useful in confirming the taxonomic identity of a biflagellated H. seosinensis gen. et sp. nov. The ITS1-5.8S-ITS2 region of Halocafeteria is the one to be characterized for the very first time in the whole family of Bicosoecida. Due to discrepancy in placing Halocafeteria in Bicosoecida family, there is need in the characterization of ITS region sequences of stramenopiles and to use multigenes (such as 18S and ITS regions) for reconstructing the evolution tree.

Keywords

• Bicosoecida
• Halocafeteria
• Halophile
• Nanoflagellate
• Protist
• Phylogeny