4.7 Article

Licorice Germplasm Resources Identification Using DNA Barcodes Inner-Variants

期刊

PLANTS-BASEL
卷 10, 期 10, 页码 -

出版社

MDPI
DOI: 10.3390/plants10102036

关键词

DNA barcoding; next-generation sequencing (NGS); single-molecule real-time (SMRT) sequencing; germplasm resources; licorice

资金

  1. National Major Science and Technology Projects [2019ZX09201005]
  2. National Key R&D Program of China [2019YFC1711100]
  3. Funds for Fostering Outstanding Scholars in Science and Technology (Innovation) [ZZ13-YQ-047]
  4. Fundamental Research Funds for the Central public welfare research institutes [ZXKT19016]
  5. TCM One Belt and One Road Project of CACMS [GH201915]
  6. APC

向作者/读者索取更多资源

A new method for determining the heterozygosity of licorice seed mixture was proposed, based on high-throughput full-length multiple DNA barcodes. The results showed that this method successfully identified the complete gradients of complex mixed samples. This method provides a brilliant and feasible way for evaluating the heterozygosity of licorice seeds.
Based on the gradual transformation from wild growth to artificial cultivation, the accurate authentication of licorice seeds contributes to the first committed step of its quality control and is pivotal to ensure the clinical efficacy of licorice. However, it is still challenging to obtain genetically stable licorice germplasm resources due to the multi-source, multi-heterozygous, polyploid, and hybrid characteristics of licorice seeds. Here, a new method for determining the heterozygosity of licorice seed mixture, based on the various sites, and finding the composition characteristics of licorice seed is preliminarily designed and proposed. Namely, high-throughput full-length multiple DNA barcodes(HFMD), based on ITS multi-copy variation exist, the full-length amplicons of ITS2, psbA-trnH and ITS are directly sequenced by rDNA through the next-generation sequence(NGS) and single-molecule real-time (SMRT) technologies. By comparing the three sequencing methods, our results proved that SMRT sequencing successfully identified the complete gradients of complex mixed samples with the best performance. Meanwhile, HFMD is a brilliant and feasible method for evaluating the heterozygosity of licorice seeds. It shows a perfect interpretation of DNA barcoding and can be applied in multi-base multi-heterozygous and polyploid species.

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