ABSTRACT
The DNA extraction method of animal medicine material is difficult and un-unified,which limits the application of molecular identification to identify animal medicines.In this study,based on the DNA extraction theory of SDS,we assessed the effects of three elements including different EDTA concentrations (0.025 mol·L-1,0.25 mol· L-1,and 0.5 mol· L-1) and whether containing NaCl and Triton X-100 in the lysis buffer on the quality of DNA extracted from different kinds of animal medicine.The optimized lysis buffer was used to extract DNA from 121 commercial animal medicines for original and species identification.The results showed that the lysis buffer of 1％ SDS,0.03 mol· L-1 Tris-HCl,0.25 mol· L-1 EDTA and 0.2 mol· L-1 NaCl had the optimum effect on DNA extraction.This lysis buffer can obtain DNA from animal medicine which is difficult to extract,such as Cicadae periostracum.The DNA extractions of 121 commercial animal medicines by optimized lysis buffer can satisfy the experimental requirements for molecular identification.All samples of commercial animal medicines can be accurately identified to the level of species.It was concluded that optimized lysis buffer can be used in the DNA extraction of different kinds of animal medicines except shells,secretions and processed products.This method provides technique support for the molecular identification of animal medicines.
ABSTRACT
As the specific endangered medicinal plant in Xinjiang, resources and distribution of Ferula sinkiangen-sis are important for biodiversity conservation and sustainable development of Chinese medicine resources. The spa-tial distribution and resources of F. sinkiangensis were researched based on low altitude remote sensing and sample investigation. The results showed that the optimum working time for F. sinkiangensis monitoring by low altitude remote sensing was the nearby 5 hills, which covered about 0.88 km2. It was suggested that the fence area should be expanded for protection. According to the results of low altitude remote sensing, the amount of F. sinkiangensis in yellow (diameter exceeding 0.3 m) was about 3 191. However, the sample investigation on amount of F. sinkiangensis in yellow (diameter exceeding 0.3 m) was about 2 752. The error between them was 14%. The monitoring time and range for F. sinkiangensis by low altitude remote sensing were also ensured. It was concluded that low altitude re-mote sensing had the advantage of quickly receiving distribution situation of F. sinkiangensis, which can effectively evaluate dynamic changes of F. sinkiangensis in Xinjiang.
ABSTRACT
Objective: This study aimed to distinguish Serpentis Periostracum from its adulterants, which will provide the basis for its safe application. Methods: Here, COI sequences of 68 samples from 13 species were PCR amplified and sequenced. Furthermore, the DNA Barcoding Gap and phylogenetic cluster analysis were carried out. Results:The results exhibited that the COI sequences of all the three origin animals of Serpentis Periostracum have DNA Barcoding Gap. For phylogenetic cluster analysis, all the three origin animals showed monophyletic and every species can be discriminated clearly. Conclusion: COI is an effective DNA barcode for the identification of Serpen-tis Periostracum.
ABSTRACT
DNA barcoding technology is widely opplied for species identification and discovery by using short and standard fragments of DNA sequences. In this study, the cytochrome c oxidase subunit 1 (COI) sequence as a DNA barcode was used to identify the Gekkogecko Linnaeus medicinal materials and adulterants in order to provide a new identification method of G. gecko Linnaeus. Genomic DNA was extracted from the experimental samples. The COI regions of nrDNA were amplified using polymerase chain reaction and sequenced bidirectionally. The alignment and NJ tree construction were performed in MEGA6.0. COI sequences can be obtained from all samples. The average intra-specific K2P distance of G. gecko Linnaeus was 0.005 and the maximum intra-specific distance was 0.013. All the G. gecko Linnaeus medicinal samples clustered into a clade in the NJ tree and can be distinguished from the adulterants. In a conclusion, COI can be used to correctly identify G. gecko Linnaeus, and it will be a potential DNA barcode for identification of other animal medicinal materials.
ABSTRACT
Medicinal animals are important part of Traditional Chinese medicine resources in China. Cytochrome c oxidase subunit I (COI) was selected as the standard DNA barcoding sequence for animal medical materials. In this study, the 51 animal species from 45 animal medical materials in the Chinese Pharmacopoeia were selected and the intra-specific variation and the inter-specific divergence, the barcoding gap, the identification efficiency of their COI sequences were analyzed. The results showed that the inter-specific divergence is higher than intra-specific distance. The barcoding gap existed between inter-specific sequence divergence and intra-specific dis-tance. The identification efficiencies were 100% both at the genus and species level except the Arthropoda. The cluster dendrogram exhibited that different species distinguished from others. Therefore, COI sequence as a bar-code is suitable to identify the species of animal medical materials in Chinese Pharmacopoeia.
ABSTRACT
matK is one of the core DNA barcode markers for plant DNA barcode identification and its universality using single makers has been in controversy. However, the universalities of different matK primer pairs in same seed plant group (order) and same matK primer pairs in different seed plant groups (order) are lack of systematic research. In this study, we collected 14563 full-length matK sequences of 11429 species of 3292 genera in 239 families belonging to 36 orders in seed plants. The universalities of 13 matK primer pairs and its 78 primer com-binations have been assessed using bioinformatics methods. The results indicated that xf/5r, 1F/8R, 390F/1326R and 3F_KIM/1R_KIM were the four most universality primer pairs. The four markers' universalities were 91.18%, 84.65%, 79.81% and 80.94% respectively in all 11429 seed plants. The most universality primer pairs in different orders were different. For each order, the primer pair with maximum universality was different. the xf/5r was the basal primer pair for primer combination and 1F/8R, 1F/1R, M3/M4 and 3F_KIM/1R_KIM could be the complementary primer pairs. This study could be a valuable resource for the primer selection of the research DNA barcoding identification in seed plants.
ABSTRACT
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.
ABSTRACT
The dried succulent stems of Cistanche (Cistanche deserticola Y. C. Ma and Cistanche tubulosa Wight.) are one of the most widely used components of traditional Chinese medicines. However, it is often confused and substituted with the roots of Orobanche pycnostachya, Boschniakia rossica (Cham. & Schltdl.) Standl., Cistanche sinensis Beck, and Cistanche salsa (C. A. Mey.) Beck. In this study, we identified psbA-trnH regions from species and tested their suitable for the identification of the above mentioned taxa. The psbA-trnH sequences showed considerable variations between species and thus were revealed as a promising candidate for barcoding of Cistanche species. Additionally, the average genetic distance of psbA-trnH ranging from 0.077% to 0.743%. In contrast, the intra-specific variation among Cistanche species was found to be significantly different from those of other species, with percentages of variation studied ranged from 0% to 0.007%. The sequence difference between the psbA-trnH sequences of Cistanche species and Orobanche pycnostachya ranged from 0.979% to 1.149%. The distance between the Cistanche species and Boschniakia rossica ranged from 1.066% to 1.224%. Our results suggest that the psbA-trnH intergenic spacer region represent a barcode that can be used to identify Cistanche species and other morphologically undistinguishable species.