Panax quinquefolius L.(American ginseng)and Panax ginseng C.A.Meyer are famous herbal medicines.Accurate authentication of the species has grown to be a significant impact.This study aims to develop a PCR kit for the ...Panax quinquefolius L.(American ginseng)and Panax ginseng C.A.Meyer are famous herbal medicines.Accurate authentication of the species has grown to be a significant impact.This study aims to develop a PCR kit for the authentication of Panax quinquefolius L.and Panax ginseng based on the nuclear internal transcribed spacer 2(ITS2)gene with the improved PCR-restriction fragment length polymorphism(PCR-RFLP)method.The reagent components in the development work were prepared and determined by the kit consisting of the DNA extraction,PCR amplification,and restriction enzyme digestion systems.A total of 21 batches of Panax quinquefolius L.and Panax ginseng samples collected from different areas were validated.Their specificity,stability,and repeatability were evaluated.The purity of genomic DNA extracted was 1.73±0.13 according to the ratio of A_(260)/A_(280),and the mass concentration was 3.15±0.22μg/g(using the kit).PCR amplicons of Panax quinquefolius L.and Panax ginseng were 122 bp in length.After the PCR products were digested by restriction enzyme Hinf I,a distinct pattern exhibited in the species of Panax quinquefolius L.with two fragments of 40 bp and 80 bp respectively,whereas those from Panax ginseng in addition to adulterated samples could not.Evaluation confirmed that the DNA kit results were stable and repeatable after 10,15,and 20 freeze-thaw cycles:the evaluation was 100%specific.The DNA kit proposed in the study can be used for the identification of Panax quinquefolius L.展开更多
Objective To make the identification of medicinal herbs in Salvia L.quickly and accurately.Methods In this work,DNA barcoding and chemical fingerprint were compared for the identification of herbs in Salvia L.First,th...Objective To make the identification of medicinal herbs in Salvia L.quickly and accurately.Methods In this work,DNA barcoding and chemical fingerprint were compared for the identification of herbs in Salvia L.First,the nucleotide sequences of the internal transcribed spacer region two amplified from 48 medicinal plants in Salvia L.,and three other groups of medicinal plants in Lamiaceae were sequenced.A molecular phylogeny was constructed using the minimum evolution and maximum parsimony methods according to their sequence diversity.Second,the water-solution bioactive components and lipid soluble components were tested by HPLC.Then a chemical phylogeny was built using HPLC fingerprint data.Comparing the molecular and chemical phylogenetic trees revealed many similarities.Results DNA barcoding was sequencing based and could therefore provide more accurate results within a shorter time especially in large-scale studies.Conclusion The results show that ITS2 region is a novel DNA barcode for the authentication of the species in Salvia L.This is the first work to show the relationship between DNA barcoding and chemical components.展开更多
基金The present project was financially funded by Jilin Provincial Department of Education,China(JJKH20180377KJ)Jilin Provincial Department of Science and Technology,China(20190304108YY,20200404152YY,20200403047SF)TCM Science and Technology Project of Jilin Province,China(2019132).
文摘Panax quinquefolius L.(American ginseng)and Panax ginseng C.A.Meyer are famous herbal medicines.Accurate authentication of the species has grown to be a significant impact.This study aims to develop a PCR kit for the authentication of Panax quinquefolius L.and Panax ginseng based on the nuclear internal transcribed spacer 2(ITS2)gene with the improved PCR-restriction fragment length polymorphism(PCR-RFLP)method.The reagent components in the development work were prepared and determined by the kit consisting of the DNA extraction,PCR amplification,and restriction enzyme digestion systems.A total of 21 batches of Panax quinquefolius L.and Panax ginseng samples collected from different areas were validated.Their specificity,stability,and repeatability were evaluated.The purity of genomic DNA extracted was 1.73±0.13 according to the ratio of A_(260)/A_(280),and the mass concentration was 3.15±0.22μg/g(using the kit).PCR amplicons of Panax quinquefolius L.and Panax ginseng were 122 bp in length.After the PCR products were digested by restriction enzyme Hinf I,a distinct pattern exhibited in the species of Panax quinquefolius L.with two fragments of 40 bp and 80 bp respectively,whereas those from Panax ginseng in addition to adulterated samples could not.Evaluation confirmed that the DNA kit results were stable and repeatable after 10,15,and 20 freeze-thaw cycles:the evaluation was 100%specific.The DNA kit proposed in the study can be used for the identification of Panax quinquefolius L.
基金supported by the International Cooperation Program of Science and Technology (No. 2007DFA30990)the Special Founding for Healthy Field (No. 200802043) awarded by the Chinese Ministry of Science and Technologysupported by grants from the Hong Kong Research Grant Council (HKU 7526/06M) to C.L.
文摘Objective To make the identification of medicinal herbs in Salvia L.quickly and accurately.Methods In this work,DNA barcoding and chemical fingerprint were compared for the identification of herbs in Salvia L.First,the nucleotide sequences of the internal transcribed spacer region two amplified from 48 medicinal plants in Salvia L.,and three other groups of medicinal plants in Lamiaceae were sequenced.A molecular phylogeny was constructed using the minimum evolution and maximum parsimony methods according to their sequence diversity.Second,the water-solution bioactive components and lipid soluble components were tested by HPLC.Then a chemical phylogeny was built using HPLC fingerprint data.Comparing the molecular and chemical phylogenetic trees revealed many similarities.Results DNA barcoding was sequencing based and could therefore provide more accurate results within a shorter time especially in large-scale studies.Conclusion The results show that ITS2 region is a novel DNA barcode for the authentication of the species in Salvia L.This is the first work to show the relationship between DNA barcoding and chemical components.
