Abstract
Background. The term “DNA barcode” is used extensively in molecular taxonomy. Basically, this technique is based on the use of a DNA sequence (about 400–800 bp) as a standard to identify and determine the species relation of organisms quickly and accurately. Therefore, DNA barcodes not only help taxonomists in classifying and identifying species, but also improve their ability to control, understand and utilize biodiversity. In this study, the authors conducted identification of samples of Anoectochilus setaceus Blume collected in Thanh Hoa through the isolated sequence of ITS gene regions.Materials and methods. Total DNA was extracted from young leaves of A. setaceus samples using CTAB method. The ITS gene segment was amplified by PCR and sequenced. This genetic sequence was analyzed, compared and used to establish a phylogenetic tree using BioEdit, BLAST and DNASTAR programs.Results and conclusion. We isolated 4 sequences of the ITS gene region in 4 A. setaceus samples collected at Xuan Lien and Pu Luong of Thanh Hoa province; the ITS gene region was 667 nucleotide long. The findings identified the samples as the same species and showed 99% similarity to the ITS gene sequence of A. roxburghii (Wall.) Lindl. published in GenBank, GQ328774. This study also demonstrates that the method employing internal transcribed spacer (ITS) sequences is an effective tool to identify A. setaceus taxa.
Highlights
In medical terms, Anoectochilus setaceus Blume is a powerful medicinal orchid which has functions beneficial for human health, i.e., improving blood circulation, producing antibacterial, anti-tracheitis, anti-hepatitis, and anti-neurasthenia effects (Lin et al, 1993; Lin et al, 2000; Wang et al, 2002; Ye et al, 2017)
internal transcribed spacer (ITS) gene amplification of A. setaceus samples After being amplified using PCR techniques, the ITS gene of A. setaceus samples were electrophoresed on 0.8% agarose gel and photographed
The findings show that the ITS gene sequences of A. setaceus samples collected at Xuan Lien (XL1 and Xuan Lien 2 Sequence (XL2)) and Pu Luong (PL1 and Pu Luong 2 ITS sequence (PL2)) and ITS gene sequence of A. roxburghii (GQ328774) were related to the same species and had a relatively large dissociation coefficient with A. lanceolatus (KT344108)
Summary
The term “DNA barcode” is used extensively in molecular taxonomy This technique is based on the use of a DNA sequence (about 400–800 bp) as a standard to identify and determine the species relation of organisms quickly and accurately. The authors conducted identification of samples of Anoectochilus setaceus Blume collected in Thanh Hoa through the isolated sequence of ITS gene regions. Total DNA was extracted from young leaves of A. setaceus samples using CTAB method. The findings identified the samples as the same species and showed 99% similarity to the ITS gene sequence of A. roxburghii (Wall.) Lindl. This study demonstrates that the method employing internal transcribed spacer (ITS) sequences is an effective tool to identify A. setaceus taxa. Исследование также демонстрирует, что использование внутреннего транскрибируемого спейсера (ITS) является эффективным методом для идентификации образцов A. setaceus.
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