Weishan Mu1, Zhaolei Zhang1,
Qing Zhao1, Weijun Kong2, Hongye
Zhao1, Yanze Liu2, Jinxin Liu1, 2*, Linchun Shi2*
1 Hebei Key Laboratory of Study and Exploitation of
Chinese Medicine, Chengde Medical University, Chengde, 067000, China
2 Institute of Medicinal Plant Development, Chinese
Academy of Medical Sciences, Peking Union Medical College, Beijing,
100193, China
* Correspondence : Jinxin Liu, liujx_23@163.com; Linchun Shi,
linchun_shi@163.com
Abstract
High-throughput
detection and screening of biological ingredients in a single bulk
sample containing a wide range of taxa still have considerable problems.
Recently, a new approach named “shotgun metabarcoding” combining
shotgun sequencing and multiple barcodes has been proposed. Here,
Shenling Baizhu San (SLBZS), a traditional patent medicine composed of
ten plant and fungus ingredients was used to test its species detection
capability. Mock and pharmaceutical samples of SLBZS were collected, and
a total of 39.52 Gb of raw data were obtained via PCR-free
shotgun sequencing. All ingredients made for mock samples can be
successfully re-detected except Coix lacryma-jobi was failed to
be detected in the second mock sample, and the positive control, the
roots of Panax quinquefolius , can be re-detected in the second
mock sample. For pharmaceutical samples, not only labeled ingredients
but also adulterants, e.g. P. quinquefolius , were detected in a
sample. The presence of P. quinquefolius was verified using the
SNP detection method with a pair of Panax specific primers. In
addition, 18 genera of fungal species were detected in both mock and
pharmaceutical samples, however, the total number of fungal reads was
relatively small. As a result, this study confirmed that shotgun
metabarcoding could authenticate all the biological ingredients of
SLBZS, has the potential to enable genomic analyses ideally of all
species a single bulk sample.
Keywords: shotgun metabarcoding, High-throughput sequencing,
multiple loci, Shenling Baizhu San, species detection,adulterant