Internal Transcribed Spacer (ITS) Fails Barcoding of the Genus Neotinea Rchb.f. (Orchidaceae)

Year 2021, Volume: 27 Issue: 1, 69 - 75, 04.03.2021

Kaan Hürkan , Kemal Melih Taşkın

https://doi.org/10.15832/ankutbd.615848

Cited By: 2

Abstract

Internal Transcribed Spacer (ITS) is one of the most used barcoding regions for the molecular phylogenetics and barcoding of orchids. Our aim in this study is to test the reliability of ITS on barcoding of closely related Neotinea spp., including Neotinea tridentata, Neotinea ustulata subsp. ustulata and Neotinea ustulata subsp. aestivalis, by comparing it to the accD-psaI intergenic spacer of the plastid DNA. Both ITS and accD-psaI regions were amplified by specific primer sets and sequenced. Phylogenetic trees were regenerated by using Maximum Parsimony approach. The results showed that ITS separated some N. tridentata samples of Turkish, Greek, Hungarian and Croatian samples from the others on the phylogenetic trees due to the incomplete lineage sorting. In contrast to ITS, the accD-psaI marker could successfully separate N. tridentata and N. ustulata samples according to a priori species classification. Our findings refer to a hybridisation story between some N. tridentata and N. ustulata. We propose not to use ITS sequences directly as a barcode and to reconstruct the phylogeny of the Neotinea group. Instead, the inclusion of other nuclear regions such as LFY, ADH, etc., or utilisation of whole genome sequencing could give better barcoding results.

Keywords

accD-psaI marker, orchids, DNA barcoding, phylogenetic incongruence

Supporting Institution

Çanakkale Onsekiz Mart University, Scientific Research Coordination Unit

Project Number

BAP FBA-2015-415

Thanks

We are grateful to Dr. Gábor Sramkó for sharing his Neotinea spp. collection. Also, we thank anonymous reviewers who helped to improve the manuscript and Cathy Seither for language proofing. This study has been financially supported by Çanakkale Onsekiz Mart University, Scientific Research Coordination Unit; Project number: BAP FBA-2015-415. This study is based on the Doctoral Thesis titled “Studies on Phylogenetic Analysis of Neotinea ustulata and Neotinea tridentata (Orchidaceae)” written by Kaan Hürkan.

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