Molecular identification of the traditional herbal medicines, Arisaematis Rhizoma and Pinelliae Tuber, and common adulterants via universal DNA barcode sequences

Abstract

ABSTRACT. Methods to identify Pinelliae Tuber and Arisaematis Rhizoma are required because of frequent reciprocal substitution between these two herbal medicines and the existence of several closely related plant materials. As a result of the morphological similarity of dried tubers, correct discrimination of authentic herbal medicines is difficult by conventional methods. Therefore, we analyzed DNA barcode sequences to identify each herbal medicine and the common adulterants at a species level. To verify the identity of these herbal medicines, we collected five authentic species (Pinellia ternata for Pinelliae Tuber, and Arisaema amurense, A. amurense var. serratum, A. erubescens, and A. heterophyllum for Arisaematis Rhizoma) and six common adulterant plant species. Maturase K (matK) and ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) genes were then amplified using universal primers. In comparative analyses of two DNA barcode sequences, we obtained 45 species-specific nucleotides sufficient to identify each species (except A. erubescens with matK) and 28 marker nucleotides for each species (except P. pedatisecta with rbcL). Sequence differences at corresponding positions of the two combined DNA barcodes provided genetic marker nucleotides that could be used to identify specimens of the correct species among the analyzed medicinal plants. Furthermore, we generated a phylogenetic tree showing nine distinct groups depending on the species. These results can be used to authenticate Pinelliae Tuber and Arisaematis Rhizoma from their adulterants and to identify each species. Thus, comparative analyses of plant DNA barcode sequences identified useful genetic markers for the authentication of Pinelliae Tuber and Arisaematis Rhizoma from several adulterant herbal materials.

Potential DNA barcodes for Melilotus species based on five single loci and their combinations

“…However, due to hybridization, it is difficult to mutate nucleotide sequences with higher conservation and synonymous substitution rates [ 60 ]. The nuclear ITS region is regarded as a core marker for identifying poisonous mushrooms [ 59 ], and our analysis confirmed that ITS is indeed a powerful potential barcode.…”

“…Such problems can be attributed to complexity arising from reproductive and evolutionary behaviour, such as hybridization, polyploidization and mixture of sexual and asexual reproduction [ 47 , 58 ]. Furthermore, in evolutionary history, introgression, reticulate evolution and incomplete lineage sorting may blur species boundaries, leading to impediments in clear barcoding [ 38 , 59 ]. However, due to hybridization, it is difficult to mutate nucleotide sequences with higher conservation and synonymous substitution rates [ 60 ].…”

Forensic genetics and genomics: Much more than just a human affair

“…These genetic markers have been applied to perform identification in a variety of food products such as olive oil [e.g., 209 , 210 ], grapevine cultivars [e.g., 211 , 212 , 213 ], composition of honey [e.g., 214 , 215 , 216 ], mushrooms [e.g., 217 , 218 , 219 ], dairy products [e.g., 220 , 221 , 222 ], seafood products [ 20 ], or meat species adulteration [ 223 ]. Additional documented cases include: i ) identification of cultivars of basmati rice [ 224 ], pome [ 225 ] and stone fruits [ 226 ], leguminosae [ 227 , 228 ], coffee [ 229 ], and tea and infusions [ 230 ]; ii ) patent misappropriation of strawberry cultivars [ 231 ]; iii ) confirmation of Protected Designation of Origin (PDO), Protected Geographical Indication (PGI), or Traditional Speciality Guaranteed (TSG) in olive [ 232 ] and grape [ 213 , 233 ] products; iv ) adulteration of traditional medicines [ 234 , 235 ] and herbs or spices [ 236 ]; v ) insufficient and erroneous food labelling, including the presence of some hidden allergens [ 237 , 238 ] or genetically modified organisms [ 239 ] (GMOs; see section Genetically modified organisms).…”

Differentiating Authentic Adenophorae Radix from Its Adulterants in Commercially-Processed Samples Using Multiplexed ITS Sequence-Based SCAR Markers

“…This method also has been used to identify the botanical origins of herbal medicines and for discriminating between authentic product and inauthentic adulterants [13,15,16]. Over 17 DNA barcode regions can be used to identify the species of Plantae and to differentiate authentic herbal medicines [1], with the genes RuBisCO (rbcL) and maturase-K (matK) in the chloroplast genome being commonly used [17,18]. The internal transcribed spacer (ITS) sequences within the nuclear ribosomal RNA genes (nrDNA) may be used to distinguish plants at the species level [19].…”