Using DNA Barcode matK and ITS in Molecular Identification of the Anoectochilus roxburghii (Wall.) Wall. ex Lindl. 1840 Plant Collected in Luang Namtha, Laos
Main Article Content
Abstract
The Anoectochilus roxburghii (Wall.) Wall. ex Lindl. 1840 species has been
over-exploited and is listed in the Red Data Book of Vietnam. In this article, we present the identification results of the species collected from Luang Namtha province, Laos, using DNA barcodes (matK and ITS). The matK gene fragment and the ITS region isolated on the agarose gel had lengths consistent with theoretical calculations, approximately 900 bp and 500 bp, respectively. The BLAST analysis of the matK gene and ITS region obtained in this study has the highest similarity - 100% and 98.39%, respectively - to Anoectochilus roxburghii and Anoectochilus burmannicus species in the GenBank. Each sequence exhibits 15 nucleotide position variations compared to the reference sequences. The results of molecular phylogenetic analysis demonstrated that the matK and ITS barcodes identified the geneus Anoectochilus, in which, ITS region can be used for differentiation of the Anoectochilus roxburghii species.
Keywords:
Anoectochilus roxburghii (Wall.) Wall. ex Lindl. 1840, DNA barcode, Laos, matK, ITS region.*
References
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https://doi.org/10.3390/app14041415.
[8] H. Li, W. Xiao, T. Tong, Y. Li, M. Zhang, X. Lin, X. Zou, Q. Wu, X. Guo, The Specific DNA Barcodes Based on Chloroplast Genes for Species Identification of Orchidaceae Plants, Scientific Reports, Vol. 11, 2021, pp. 1424.
[9] H. T. T. Hue, N. T. B. Ngoc, D. Q. Ha, L. T. T. Hien, N. X. Canh, Analysis of DNA Barcode Region ITS and matK in some Polyscias plants of Vietnam, TNU Journal of Science and Technology, Vol. 227, No. 5, 2022, pp. 318-325.
[10] T. Yang, T. L. Zhang, Y. H. Guo, X. Liu, Identification of Hybrids in Potamogeton: Incongruence between Plastid and ITS Regions Solved by a Novel Barcoding Marker PHYB, PLOS ONE, Vol. 11, No. 11, 2016, pp. e0166177.
[11] B. T. M. Huong, N. V. Phong. Study on Identification of DNA Barcode Sequence of Liriodendron Chinense to Identify Plant Species, Journal of Forestry Science and Technology, Vol. 5, 2020, pp. 012-021.
[12] N. H. Manh, L. T. T. Hang, N. T. H. Mai, N. T. P. Trang, N. V. Sinh, Molecular Characteristic of RbcL and trnH-psbA of Abies delavayi subsp. fansipanensis (Xiang Q.P.) Rurhforth in Vietnam, Vietnam Journal of Science and Technology, Vol. 63, No. 3, 2021, pp. 28-32.
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[14] P. T. T Nhan, L. P Dung, P. T. N. Khanh, T. Xaykham, C. H. Mau, DNA Barcode of MatK Combined with ITS Effectively Distinguishes the Medicinal Plant Stephania Brachyandra Diels Collected in Laocai, Vietnam, Journal of Applied Biology and Biotechnology, Vol. 9, No. 6, 2021, pp. 63-70.
[15] K. Tamura, M. Nei. Estimation of the Number of Nucleotide Substitutions in the Control Region of Mitochondrial DNA in Humans and Chimpanzees, Molecular Biology and Evolution, Vol.10, 1993, pp. 512-526.
[16] J. Felsenstein, Confidence Limits on Phylogeneies: An Approach using the Bootstrap, Evolution, Vol. 39, 1985, pp. 783-791.
[17] S. Kumar, G. Stecher, M. Li, C. Knyaz, K. Tamura. MEGA X: Molecular Evolutionary Genetics Analysis Across Computing Platforms, Molecular Biology and Evolution, Vol. 35, 2018, pp. 1547-1549.
[18] L. T. M. Thu, T. T. Thuy, C. H. Mau, The Species Identification with DNA Barcode and the Sequence Analysis of ITS and RpoC1 of Anoectochilus Sample Collected at Thuan Chau, Son La, Vietnam, Journal of Science and Technology - Thai Nguyen University, Vol. 9, 2019, pp. 107-114.
[19] H. H. Duc, N. T. Giang, D. H. Xo, H. T. Loan, P. D. Yen, T. T. Tuan, D. D. Giap, Research on using some DNA Barcodes in Genetic Analysis and Identification of some Anoectochilus Spp. Plants, Journal of Science - Can Tho University, Vol. 55, No. 1, 2019, pp. 14-23.
[20] H. V. The, T. T. K Phuong, V. T. T. Tram, S. Widiarsih, Comparison of matK and rbcL DNA Barcodes for Genetic Classification of Jewel Orchid Accessions in Vietnam, Journal of Genetic Engineering and Biotechnology, Vol. 19, No. 1, 2021, pp. 93.