title: Authentication of official Da-huang by sequencing and multiplex allele-specific PCR of a short maturase K gene.
abstract: Rhubarb (official Da-huang) is an important medicinal herb in Asia. Many adulterants of official Da-huang have been discovered in Chinese markets in recent years, which has resulted in adverse effects in medicinal treatment. Here, novel molecular markers based on a short maturase K (matK) gene were developed for authenticating official Da-huang. This study showed that all the species from official Da-huang were clustered together in one clade in the polygenetic trees based on short matK. Two highly conserved single nucleotide polymorphisms of short matK were mined in the species from official Da-huang. Based on these polymophisms, four improved specific primers of official Da-huang were successfully developed that generated reproducible specific bands. These results suggest that the short matK sequence can be considered as a favorable candidate for distinguishing official Da-huang from its adulterants. The established multiplex allele-specific PCR was determined to be simple and accurate and may serve as a preferable tool for authentication of official Da-huang. In addition, we suggest that short-sized specific bands be developed to authenticate materials used in traditional Chinese medicine.
authors.affiliation: School of Chinese Pharmacy, Beijing University of Chinese Medicine, No. 6, WangJing ZhongHuan South Street, Chao-Yang District, Beijing 100102, PR China.
authors.forename: Guojie
authors.initials: G
authors.lastname: Xu
authors.forename: Xueyong
authors.initials: X
authors.lastname: Wang
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Liu
authors.forename: Weidong
authors.initials: W
authors.lastname: Li
authors.forename: Shengli
authors.initials: S
authors.lastname: Wei
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Cheng
authors.forename: Juan
authors.initials: J
authors.lastname: Liu
mesh_terms: D000483:Alleles; D004365:Drugs, Chinese Herbal / Q000737:chemistry; D004722:Endoribonucleases / Q000032:analysis / Q000235:genetics*; D017343:Genes, Plant; D005819:Genetic Markers; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009713:Nucleotidyltransferases / Q000032:analysis / Q000235:genetics*; D010802:Phylogeny; D010940:Plant Proteins / Q000032:analysis / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D012250:Rheum / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D017422:Sequence Analysis, DNA
chemical_list: C521634:Dahuang Zhechong pill; D004365:Drugs, Chinese Herbal; D005819:Genetic Markers; D010940:Plant Proteins; D009713:Nucleotidyltransferases; C034038:mRNA maturase; D004722:Endoribonucleases