One of the difficulties encounter in the selection of the correct plant material is to establish the identity of certain species that may be known by different binomial botanical names in different regions. Certain rare and expensive medicinal plant species are often adulterated or substituted by mo...
One of the difficulties encounter in the selection of the correct plant material is to establish the identity of certain species that may be known by different binomial botanical names in different regions. Certain rare and expensive medicinal plant species are often adulterated or substituted by morphologically similar, easily available or less expensive species. DNA-based techniques have been widely used for authentication of plant species of medicinal importance. This is especially useful in case of those that are frequently substituted or adulterated with other species or varieties that are morphologically and/or phytochemically indistinguishable. Recently, many molecular marker methods have been introduced, but not enough marker research is processing in Ligusticum, Cimicifuga and Anemarrhena genus compared with other medicinal plants. In this study, we aim the development and the application of molecular markers for rapid, efficient, and reliable screening and identification of Ligusticum, Cimicifuga and Anemarrhena genus from fresh and dried organ samples. DNA markers are reliable for informative polymorphisms as the genetic composition, and it is unique to differentiate each species without any affection by physiological and environmental factors. The oriental medicinal plant Ligusticum tenuissimum (Korean name “Go-Bon”) is widely used in Korea and China. L. tenuissimum (Go-Bon) has been treated for headache, common cold and fever remedy. The internal transcribed spacer (ITS) region was sequenced from thirty-four “Go-Bon” samples collected from botanical gardens and markets in Korea and China to identify and authenticate the L. tenuissimum. Based on the ITS sequences, the thirty-four Go-Bon samples were classified into three groups: L. tenuissimum (Korean Go-Bon), L. jeholense (Chinese Go-Bon) and unknown Chinese Ligusticum species. Three specific primers were designed to identify the three groups of Ligusticum species using multiplex PCR. The established multiplex-PCR was proved to be effective for the differentiation of L. tenuissimum in commercial materials. Expensive herbs like oriental medicinal plants are always possible targets for fraudulent labeling or misuse in many countries. In the genus Cimicifuga, most species have been used as “Sheng-Ma” for oriental medicinal purposes. Among the Cimicifuga species, C. heracleifolia, C. dahurica, C. foetida, and C. simplex were used as certified medicinal materials in Korea, China and Japan. To identify these four Cimicifuga species, the internal transcribed spacer (ITS) was used for polymerase chain reaction (PCR) analysis. Nucleotide polymorphisms have been identified among four Cimicifuga species. Four specific primers were designed from their polymorphic sites to identify Cimicifuga species using multiplex PCR. The established multiplex-PCR was proved to be effective for the differentiation of Cimicifuga species in commercial materials. Anemarrhena asphodeloides Bunge (Jimo) is one of the most popular and valuable plant species in many countries, including China, Korea and Japan. The current commercial products such as Belamcanda chinensis Leman (Sagan), Paeonia albiflora Pall (Chog-chag-yag) and Peucedanum japonicum Thunberg (Shig-ban-pung) which are most similar to Jimo roots, were used for more reliable authentication method. Polymerase chain reaction (PCR) analysis of the trnL-F region has been proved to be an appropriate method for the identification of species in the A. asphodeloides genus. A single nucleotide polymorphism (SNP) has been identified in Jimo, Sagan, Chog-chag-yag and Shig-ban-pung. The specific PCR primers were designed from the SNP to differentiate the A. asphodeloides (Jimo) from Belamcanda chinensis (Sagan), Paeonia albiflora (Chog-chag-yag), and Peucedanum japonicum (Shig-ban-pung) via multiplex PCR. The established multiplex-PCR method for the rapid detection of the Jimo in a single reaction was determined to be effective for the differentiation of Jimo (A. asphodeloides). We therefore present an effective method for the genetic identification of A. asphodeloides. All of the results in this study will be helpful for the identification of Ligusticum, Cimicifuga and Anemarrhena genus. This is the first report of species-specific marker development in Ligusticum genus; L. tenuissimum, L. jeholense and unknown Ligusticum species: Cimicifuga genus; C. heracleifolia, C. dahurica, C. foetida, C. simplex and Anemarrhena asphodeloides.
One of the difficulties encounter in the selection of the correct plant material is to establish the identity of certain species that may be known by different binomial botanical names in different regions. Certain rare and expensive medicinal plant species are often adulterated or substituted by morphologically similar, easily available or less expensive species. DNA-based techniques have been widely used for authentication of plant species of medicinal importance. This is especially useful in case of those that are frequently substituted or adulterated with other species or varieties that are morphologically and/or phytochemically indistinguishable. Recently, many molecular marker methods have been introduced, but not enough marker research is processing in Ligusticum, Cimicifuga and Anemarrhena genus compared with other medicinal plants. In this study, we aim the development and the application of molecular markers for rapid, efficient, and reliable screening and identification of Ligusticum, Cimicifuga and Anemarrhena genus from fresh and dried organ samples. DNA markers are reliable for informative polymorphisms as the genetic composition, and it is unique to differentiate each species without any affection by physiological and environmental factors. The oriental medicinal plant Ligusticum tenuissimum (Korean name “Go-Bon”) is widely used in Korea and China. L. tenuissimum (Go-Bon) has been treated for headache, common cold and fever remedy. The internal transcribed spacer (ITS) region was sequenced from thirty-four “Go-Bon” samples collected from botanical gardens and markets in Korea and China to identify and authenticate the L. tenuissimum. Based on the ITS sequences, the thirty-four Go-Bon samples were classified into three groups: L. tenuissimum (Korean Go-Bon), L. jeholense (Chinese Go-Bon) and unknown Chinese Ligusticum species. Three specific primers were designed to identify the three groups of Ligusticum species using multiplex PCR. The established multiplex-PCR was proved to be effective for the differentiation of L. tenuissimum in commercial materials. Expensive herbs like oriental medicinal plants are always possible targets for fraudulent labeling or misuse in many countries. In the genus Cimicifuga, most species have been used as “Sheng-Ma” for oriental medicinal purposes. Among the Cimicifuga species, C. heracleifolia, C. dahurica, C. foetida, and C. simplex were used as certified medicinal materials in Korea, China and Japan. To identify these four Cimicifuga species, the internal transcribed spacer (ITS) was used for polymerase chain reaction (PCR) analysis. Nucleotide polymorphisms have been identified among four Cimicifuga species. Four specific primers were designed from their polymorphic sites to identify Cimicifuga species using multiplex PCR. The established multiplex-PCR was proved to be effective for the differentiation of Cimicifuga species in commercial materials. Anemarrhena asphodeloides Bunge (Jimo) is one of the most popular and valuable plant species in many countries, including China, Korea and Japan. The current commercial products such as Belamcanda chinensis Leman (Sagan), Paeonia albiflora Pall (Chog-chag-yag) and Peucedanum japonicum Thunberg (Shig-ban-pung) which are most similar to Jimo roots, were used for more reliable authentication method. Polymerase chain reaction (PCR) analysis of the trnL-F region has been proved to be an appropriate method for the identification of species in the A. asphodeloides genus. A single nucleotide polymorphism (SNP) has been identified in Jimo, Sagan, Chog-chag-yag and Shig-ban-pung. The specific PCR primers were designed from the SNP to differentiate the A. asphodeloides (Jimo) from Belamcanda chinensis (Sagan), Paeonia albiflora (Chog-chag-yag), and Peucedanum japonicum (Shig-ban-pung) via multiplex PCR. The established multiplex-PCR method for the rapid detection of the Jimo in a single reaction was determined to be effective for the differentiation of Jimo (A. asphodeloides). We therefore present an effective method for the genetic identification of A. asphodeloides. All of the results in this study will be helpful for the identification of Ligusticum, Cimicifuga and Anemarrhena genus. This is the first report of species-specific marker development in Ligusticum genus; L. tenuissimum, L. jeholense and unknown Ligusticum species: Cimicifuga genus; C. heracleifolia, C. dahurica, C. foetida, C. simplex and Anemarrhena asphodeloides.
주제어
#Molecular Markers Angelicae Tenuissimae Radix Cimicifuga Rhizome Anemarrhena Rhizome
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