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DNA barcodes for the identification of Anoectochilus roxburghii and its adulterants

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Abstract

Main conclusion

Chinese medicinal herbs have a similar appearance and are easily confused, complicating identification via traditional methods. This study provided a scientific approach, based on DNA barcoding, to accurately and rapidly identify Anoectochilus roxburghii and its adulterants. This technology complements traditional methods of identification of medicinal herbs.

A comparison of the DNA barcodes matK, psbA-trnH and ITS2 was performed to verify that the ITS2 sequence is an effective marker for rapidly and accurately identifying A. roxburghii and its closely related species. Genomic DNA extracted from A. roxburghii and its adulterants were used as templates and the ITS2 sequence was amplified using PCR amplification and sequencing. Species identification was conducted using BLAST1 and neighbor-joining trees. The 12 samples were successfully classified into four species based on the ITS2 sequence. The ITS2 sequence length of A. roxburghii was 253 bp. The average intra-specific genetic distance of A. roxburghii was 0.0021, markedly lower than the inter-specific genetic distance between A. roxburghii and its adulterants (0.0380). Our findings illustrate that ITS2 sequence can accurately and efficiently distinguish A. roxburghii and its adulterants. In addition, the results provided reference for molecular identification of other Chinese herbal medicine.

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Abbreviations

ITS2:

Internal transcribed spacer 2

PCR:

Polymerase chain reaction

TBE:

Tris-borate-EDTA

NJ:

Tree neighbor-joining tree

TLC:

Thin layer chromatography

HMM:

Hidden Markov model

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 81303167), the Zhejiang Provincial Natural Science Foundation (Grant No. LY13H280010, LY14H280008), Zhejiang Province Public Technology Applied Research Projects (Grant No. 2014C32017), Ningbo Agricultural Science and Technology Research Projects (Grant No. 2014C10013) and Science and Technology Innovation Program for Students of Zhejiang (Xinmiao Talent Scheme) (2015R412003). We thank the anonymous reviewers for carefully reviewing our manuscript and making many valuable suggestions.

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Authors and Affiliations

  1. The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang A & F University, Hangzhou, 311300, China

    Tongwei Lv, Renda Teng, Qingsong Shao, Hongzhen Wang & Lili Zhang

  2. Modern Agricultural Research Center in Ningbo, Industrial Technology Research Institute of Zhejiang University, Ningbo, 315033, China

    Wangshu Zhang

  3. Zhejiang Rare Herbal Medicine and Engineering Institute, Wuyi, 321200, China

    Mingyan Li

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  1. Tongwei Lv
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  2. Renda Teng
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  3. Qingsong Shao
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  4. Hongzhen Wang
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  5. Wangshu Zhang
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  6. Mingyan Li
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  7. Lili Zhang
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Correspondence to Qingsong Shao.

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Lv, T., Teng, R., Shao, Q. et al. DNA barcodes for the identification of Anoectochilus roxburghii and its adulterants. Planta 242, 1167–1174 (2015). https://doi.org/10.1007/s00425-015-2353-x

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  • DOI: https://doi.org/10.1007/s00425-015-2353-x

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