Molecular Authentication of the Traditional Chinese Medicinal Plant Euphorbia pekinensis

Abstract

The ITS regions of Euphorbia pekinensis and six other Euphorbia species used as adulterants of E. pekinensis were sequenced to differentiate them. The sequences are identical among the individuals in the seven species studies. Diversity in DNA sequences among various species was found ranging from 8.3 % to 43.8 % in ITS1 and 7.6 % to 36.6 % in ITS2 region. Furthermore, based on the divergent ITS regions, species-specific primers, JDJp 1 and JDJp 2, were designed in the polymorphic regions of E. pekinensis to distinguish it from adulterants. These ITS-derived primers amplified a 281-bp-specific DNA fragment from E. pekinensis. No amplified product was observed using DNA of six adulterants.

References

• 1 The State Pharmacopoeia Commission of the P RC. Pharmacopoeia of the People’s Republic of China. Beijing; Chemical Industry Press 2000
• 2 Lum M R, Potter E, Dang T, Heber D, Hardy M, Hirsch A M. Identification of botanicals and potential contaminants through RFLP and sequencing.  Planta Med. 2005;  71 841-6
• 3 Pusch C M, Giddings I, Scholz M. Repair of degraded duplex DNA from prehistoric samples using Escherichia coli DNA polymerase I and T4 DNA ligase.  Nucleic Acids Res. 1998;  26 57-9
• 4 Baldwin B G, Sanderson M J, Porter J M, Wojciechowski M F, Campbell C, Donoghue M J. The ITS region of nuclear ribosomal DNA: a valuable source of evidence on angiosperm phylogeny.  Ann Mo Bot Gard. 1995;  82 247-77
• 5 Tai-Wai Lau D, Shaw P C. Authentication of medicinal Dendrobium species by the internal transcribed spacer of ribosomal DNA.  Planta Med. 2001;  67 456-60
• 6 Thompson J D, Gibson T J, Plewniak F, Jeanmougin F, Higgins D G. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.  Nucleic Acids Res. 1997;  24 4876-82
• 7 Kumar S, Tamura K, Nei M. MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment.  Brief Bioinform. 2004;  5 150-63