Comparison and Identification of Secondary Structure of ITS2 Sequence in Bletillae Rhizoma and Its Counterfeits

CHEN Mei-jun; LI Feng-qing; LYU Meng; LIU Jia-yu; CHEN Hong-ping; LIU You-ping

doi:10.13422/j.cnki.syfjx.2017150046

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Comparison and Identification of Secondary Structure of ITS2 Sequence in Bletillae Rhizoma and Its Counterfeits

更新时间：2024-01-04

- Comparison and Identification of Secondary Structure of ITS2 Sequence in Bletillae Rhizoma and Its Counterfeits
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 15, Pages: 46-52(2017)
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- DOI：10.13422/j.cnki.syfjx.2017150046
  CLC： S567.239
- Published：2017
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CHEN Mei-jun, LI Feng-qing, LYU Meng, et al. Comparison and Identification of Secondary Structure of ITS2 Sequence in Bletillae Rhizoma and Its Counterfeits[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(15): 46-52.
DOI：

CHEN Mei-jun, LI Feng-qing, LYU Meng, et al. Comparison and Identification of Secondary Structure of ITS2 Sequence in Bletillae Rhizoma and Its Counterfeits[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(15): 46-52. DOI： 10.13422/j.cnki.syfjx.2017150046.

摘要

目的：利用DNA条形码技术通过ITS2序列对白及及其混伪品进行快速、准确鉴别。方法：采用植物基因组DNA提取试剂盒提取白及及其混伪品基因组DNA，对全部收集样品的ITS2序列进行PCR扩增和测序，所得序列经Codon Code Aligner 拼接后，利用MEGA7.0 软件进行数据分析计算物种种间种内Kimura2-parameter（K2P）遗传距离。采用最近距离法及基于ITS2序列构建Neighbor-joining（NJ）系统聚类树，结合ITS2序列二级结构相比对的方法鉴别分析。结果：K2P遗传距离及NJ聚类树均显示只能鉴别出白及属与其他科属物种，区分不开白及属内3个物种，结合ITS2二级结构进一步鉴定，可将白及属的各个种及其他科属混伪品准确地鉴别出来。结论：利用ITS2序列作为DNA条形码，结合ITS2序列的二级结构可以有效地鉴定白及及其混伪品。

Abstract

Objective: To fast and accurately identify ITS2 sequences of Bletillae Rhizoma and its counterfeits using DNA barcode technology. Method: The genomic DNA was extracted by plant genomic DNA extraction kit

and the ITS2 sequences of all collected samples were amplified and measured. The sequences were spliced by Codon Code Aligner and analyzed by MEGA7.0 software to calculate interspecies Kimura2-parameter (K2P) genetic distance. The neighbor-joining (NJ) system clustering tree was constructed by using the nearest distance method and ITS2 sequences

and the method was compared with the ITS2 sequences. Result: K2P genetic distance and NJ clustering tree only identified Bletillae Rhizoma from other species

but failed to distinguish Bletillae Rhizoma and the other three in the genus. The secondary structure of ITS2 could be used to further accurately identify Bletillae Rhizoma and other species. Conclusion: ITS2 sequences can be used as DNA bar code

and combined with the secondary structure of ITS2 sequence to effectively identify Bletillae Rhizoma and its counterfeits.

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