Identification And Application Of Chinese Herbal Seed And Seedling Based On DNA Barcoding Technology

Accurate identification of germplasm resources of Chinese herbal medicines has a great significance on preservation of national medicinal plant germplasm and improvement of medica material quality. However, there are rare methods concerning on identification of germplasm resources. In this study, we have established the methodology of identification in germplasm resources of Chinese herbal medicines based on DNA barcoding technology, combined with morphological method. We initiated our research with 56 Alismatis Rhizoma seeds. The results indicated that the morphological characteristics of Alismaplantago-aquatica and Alisma orientale seeds were similar. The ITS2 sequence lengths of two species were 311 bp. The GC content was 56.9%. One stable SNP (T-A) for two species was detected at 165 bp in the ITS2 sequences. Then,98 seed samples of Chinese herbal medicines from 10 species were collected and investigated. Some seed samples were identified as Paeonia lactiflora, Toxicodendron vernicifluum, Stellaria dichotoma, Caesalpinia sappan, Abutilon theophrasti, Mosla chinensis, Sarcandra glabra and Plantago asiatica based on ITS2 sequence, while the other seed samples of Phytolaccae Radix were identified as Phytolacca americana based on psbA-trnH sequences. The seed samples of Viticis Negundo Folium were identified as Vitex agnus-castus or Vitex negundo by ITS2 sequences and as Vitex negundo var. cannabifolia or Vitex negundo by psbA-trnH sequences. Furthermore, the seedlings of Dichroa febrifuga as representative were determined. The sampling regions of D. febrifuga were selected according to analysis of TCMGIS software. There are 4 variable sites was detected in the ITS2 sequences. The DNA barcoding technology can be used as a powerful tool to identify D. febrifuga and their adulterants samples. In addition, the study on classic square medicine and clinical samples based on DNA barcoding technology has confirmed a robust role in supervision of drug production and hospital administration. This research should not only contribute to the preservation of germplasm resources and the supervision of Chinese herbal medicine seeds, but also provide a model for Chinese herbal seeds and seedlings using DNA barcoding technology.