SNP, ARMS and SSH authentication of medicinal Dendrobium officinale KIMURA et MIGO and application for identification of Fengdou drugs.
Dried stems of Dendrobium officinale have been used as crude drugs in traditional Chinese medicine (TCM) with good tonic efficacy. Sequences of chloroplast, nuclear and mitochondria genes and the method of genomic DNA (gDNA) suppression subtraction hybridization (SSH) were used to authenticate different populations during the process of good agriculture practice (GAP) and crude drug quality control. Six populations could be authenticated successfully by nine single sucleotide polymorphism (SNP) sites and six pairs of diagnostic primers for amplification refractory mutation system (ARMS) were also designed to identify six populations on the basis of single nucleotide polymorphism (SNPs). The remainder two populations (JSR, GGL) with the same sequences could be authenticated by SSH. One population-specific fragment was obtained by SSH and a pair of specific primers (SSH-JB01, SSH-JB02) on the specific sequence was designed to authenticate GGL population from the other populations tested. As the resultants were population-specific, the botanic origins of fifty "Fengdou" drug samples from markets could be classified. It is evident that the combined methods provide a high throughput and reliable approach for identification of D. officinale plants and "Fengdou" drugs.
Ding
Ge
G
Jiangsu Provicinal Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University, No. 1, Wenyuan Road, Nanjing 210046, China.
Zhang
Daizhen
D
Feng
Zhenyu
Z
Fan
Wenjing
W
Ding
Xiaoyu
X
Li
Xuexia
X
D003001:Cloning, Molecular; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics; D031667:Dendrobium / Q000737:chemistry* / Q000235:genetics; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000592:standards; D007438:Introns / Q000235:genetics; D008928:Mitochondria / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D009693:Nucleic Acid Hybridization; D020641:Polymorphism, Single Nucleotide
D017931:DNA Primers; D018744:DNA, Plant; D004365:Drugs, Chinese Herbal