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. 2017 Dec 11;7:17316. doi: 10.1038/s41598-017-17695-2

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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A schematic diagram illustrating how the ITS2 sequence-divergence threshold improves species identification accuracy when using DNA barcoding. (a). An unknown sample is successfully assigned into its conspecific clade when the DNA barcoding library was complete. (b1). The unknown sample is assigned into an alternative clade when the DNA barcoding library was incomplete and could easily be mistakenly identified. (b2). K2P genetic distances between species are calculated from b1. The sequence-divergence values between the unknown sample and L. congestiflora (4.79%) is above the ITS2 AGDS threshold (3.98%), indicating that they are heterospecific.