Licorice Germplasm Resources Identification Using DNA Barcodes Inner-Variants

Abstract

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.

Perspectives of Licorice Production in Harsh Environments of the Aral Sea Regions

“…Chloroplast genomes of G. glabra [ 47 ] and G. lepidota [ 48 ] have been presented. Barcodes for identification of licorice germplasm resources [ 49 ] and SSR markers in different interspecific-cross populations [ 50 ] have been developed. However, molecular breeding of superior licorice varieties still needs more wild licorice resources as well as molecular markers for elite traits.…”