1.Specific DNA barcodes, germplasm resources, and genetic diversity of Eleutherococcus senticosus.
Zhi-Fei ZHANG ; Zhong-Zhen ZHAO ; Xin WANG ; Guang-Yao YIN ; Ying CHEN ; Jin-Hui MAN ; Yue SHI ; Yu-Ying HUANG ; Shan-Hu LIU ; Zi-Qi LIU ; Xiao-Hui WANG ; Sheng-Li WEI
China Journal of Chinese Materia Medica 2023;48(5):1229-1237
Eleutherococcus senticosus is one of the Dao-di herbs in northeast China. In this study, the chloroplast genomes of three E. senticosus samples from different genuine producing areas were sequenced and then used for the screening of specific DNA barcodes. The germplasm resources and genetic diversity of E. senticosus were analyzed basing on the specific DNA barcodes. The chloroplast genomes of E. senticosus from different genuine producing areas showed the total length of 156 779-156 781 bp and a typical tetrad structure. Each of the chloroplast genomes carried 132 genes, including 87 protein-coding genes, 37 tRNAs, and 8 rRNAs. The chloroplast genomes were relatively conserved. Sequence analysis of the three chloroplast genomes indicated that atpI, ndhA, ycf1, atpB-rbcL, ndhF-rpl32, petA-psbJ, psbM-psbD, and rps16-psbK can be used as specific DNA barcodes of E. senticosus. In this study, we selected atpI and atpB-rbcL which were 700-800 bp and easy to be amplified for the identification of 184 E. senticosus samples from 13 genuine producing areas. The results demonstrated that 9 and 10 genotypes were identified based on atpI and atpB-rbcL sequences, respectively. Furthermore, the two barcodes identified 23 genotypes which were named H1-H23. The haplotype with the highest proportion and widest distribution was H10, followed by H2. The haplotype diversity and nucleotide diversity were 0.94 and 1.82×10~(-3), respectively, suggesting the high genetic diversity of E. senticosus. The results of the median-joining network analysis showed that the 23 genotypes could be classified into 4 categories. H2 was the oldest haplotype, and it served as the center of the network characterized by starlike radiation, which suggested that population expansion of E. senticosus occurred in the genuine producing areas. This study lays a foundation for the research on the genetic quality and chloroplast genetic engineering of E. senticosus and further research on the genetic mechanism of its population, providing new ideas for studying the genetic evolution of E. senticosus.
DNA Barcoding, Taxonomic
;
Eleutherococcus/genetics*
;
Base Sequence
;
Chloroplasts/genetics*
;
Genetic Variation
;
Phylogeny
2.Comparison between macroscopic identification and DNA barcoding identification of Amomi Fructus.
En-Ai ZHAI ; Wen-Juan MI ; Yang CUI ; Wei-Feng HONG ; Ya-Shun WANG ; Xing-Yu GUO ; Hui-Qin ZOU ; Yong-Hong YAN
China Journal of Chinese Materia Medica 2022;47(17):4600-4608
This study aims to explore the consistency between macroscopic identification and DNA barcoding identification of Amomi Fructus. With the DNA barcoding identification results, we evaluated the reliability of identifying Amomi Fructus quality by combining macroscopic traits with main volatile chemical components. Thirteen batches of Amomi Fructus samples were collected for identification. Firstly, the morphological and sensory characteristics of each sample were observed and recorded according to the standard in Chinese Pharmacopoeia(2020 edition). The 100-fruit weight, longitudinal diameter, transverse diameter, and longitudinal diameter-to-transverse diameter ratio were measured, which correspond to large, solid, and full kernel representing good quality in the sensory evaluation. The odor value detected by electronic nose and major volatile components(borneol, camphor, limonene, and borneol acetate) correspond to the sensory evaluation of strong odor representing good quality. Secondly, DNA barcoding was employed to identify the 13 batches of samples. Finally, clustering analysis was performed for the main volatile components and macroscopic traits, and the identification results were compared with those of DNA barcoding. Except two batches of samples(No.6 and No.10), the macroscopic identification showed the results consistent with those of DNA barcoding, with an identification rate of 84.62%. The clustering results of the content of four volatile chemical components and macroscopic traits were also consistent with the DNA barcoding identification results. DNA barcoding can verify the results of macroscopic identification and provide a scientific basis for the inheritance and development of macroscopic identification. Moreover, the combination of macroscopic traits and chemical components demonstrates higher accuracy in the quality evaluation of Chinese medicinal materials.
Camphanes
;
Camphor/analysis*
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DNA Barcoding, Taxonomic
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Drugs, Chinese Herbal/chemistry*
;
Fruit/genetics*
;
Limonene/analysis*
;
Reproducibility of Results
3.Identification and quality evaluation of germplasm resources of commercial Scutellaria baicalensis based on DNA barcode and HPLC.
Jing-Han ZHANG ; Shan-Hu LIU ; Zhi-Fei ZHANG ; Yue SHI ; Jin-Hui MAN ; Guang-Yao YIN ; Xin WANG ; Feng-Bo LIU ; Xiao-Hui WANG ; Sheng-Li WEI
China Journal of Chinese Materia Medica 2022;47(7):1814-1823
Scutellaria baicalensis is a commonly used Chinese medicinal herb. In this study, we identified the germplasm resources of commercial S. baicalensis samples based on trnH-psbA, petA-psbJ, and ycf4-cemA sequences according to the available chloroplast genome sequencing results, and measured the content of baicalin by HPLC. Through the above means we determined the best DNA barcode that can be used to detect the germplasm resources and evaluate the quality of commercial S. baicalensis samples. A total of 104 samples were collected from 24 provinces, from which DNA was extracted for PCR amplification. The amplification efficiencies of trnH-psbA, petA-psbJ, and ycf4-cemA sequences were 100%, 59.62%, and 25.96%, respectively. The results of sequence analysis showed that 5, 4, and 2 haplotypes were identified based on trnH-psbA, petA-psbJ, and ycf4-cemA sequences, respectively. However, the sequences of haplotypes in commercial samples were different from that of the wild type, and the joint analysis of three fragments of S. baicalensis only identified 6 haplotypes. Furthermore, the phylogenetic analysis and genetic distance analysis indicated that trnH-psbA could be used to identify S. baicalensis from adulterants. The above analysis showed that trnH-psbA was the best fragment for identifying the germplasm resources of commercial S. baicalensis samples. We then analyzed the haplotypes(THap1-THap5) of commercial S. baicalensis samples based on trnH-psbA and found that THap2 was the main circulating haplotype of the commercial samples, accounting for 86.55% of the total samples, which indicated the scarce germplasm resources of commercial S. baicalensis samples. The content of baicalin in all the collected commercial S. baicalensis samples exceeded the standard in Chinese Pharmacopoeia and had significant differences(maximum of 12.21%) among samples, suggesting that the quality of commercial S. baicalensis samples varied considerably. However, there was no significant difference in baicalin content between different provinces or between different haplotypes. This study facilitates the establishment of the standard identification system for S. baicalensis, and can guide the commercial circulation and reasonable medication of S. baicalensis.
Chromatography, High Pressure Liquid
;
DNA Barcoding, Taxonomic/methods*
;
DNA, Plant/genetics*
;
Phylogeny
;
Scutellaria baicalensis/genetics*
4.Molecular identification and efficacy analysis of herbs at Orussey Herbal Market, Phnom Penh, Cambodia.
Xue-Ping WEI ; Yu-Qing DONG ; Ting-Yan QIANG ; Wen-Jie LI ; Yi-Chen SONG ; Ben-Gang ZHANG ; Zhao ZHANG ; Theang HUOT ; Hai-Tao LIU ; Yao-Dong QI
China Journal of Chinese Materia Medica 2021;46(24):6312-6322
Cambodia is rich in medicinal plant resources. One hundred and thirty-three medicinal material samples, including the hole herb, root, stem/branch, leaf, flower, fruit, seed, and resin, were collected from the Orussey Herbal Market in Phnom Penh, Cambodia, and then authenticated by ITS and psbA-trnH. A total of 46 samples were identified based on ITS sequences, belonging to 24 families, 40 genera, and 42 species. A total of 100 samples were identified by psbA-trnH sequences to belong to 42 families, 77 genera, and 84 species. A total of 103 samples were identified by two DNA barcodes. According to the morphological characteristics of the medicinal materials, 120 samples classified into 50 species, 86 genera, and 86 families were identified, and the majority of them were from Zingiberaceae, Fabaceae, and Acanthaceae. Such samples have been commonly used in traditional Cambodian medicine, Ayurvedic medicine, Unani medicine, traditional Chinese medicine, and ethnomedicine, but different medical systems focus on different functional aspects of the same medicinal material. The results of this study have demonstrated that DNA barcoding has a significant advantage in identifying herbal products, and this study has provided basic data for understanding the traditional medicinal materials used in Cambodia.
Cambodia
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DNA Barcoding, Taxonomic
;
DNA, Plant/genetics*
;
Humans
;
Plant Leaves
;
Plants, Medicinal/genetics*
5.DNA barcoding identification of original plants of a rare medicinal material Resina Draconis and related Dracaena species.
Yue ZHANG ; Mei-Fang SONG ; Hai-Tao LI ; Hui-Fang SUN ; Zhong-Lian ZHANG
China Journal of Chinese Materia Medica 2021;46(9):2173-2181
Resina Draconis, a rare and precious traditional medicine in China, is known as the "holy medicine for promoting blood circulation". According to the national drug standard, it's derived from the resin extracted from the wood of Dracaena cochinchinensis, a Liliaceae plant. In addition, a variety of Dracaena species all over the world can form red resins, and there is currently no molecular identification method that can efficiently identify the origin of Dracaena medicinal materials. In this study, seven species of Dracaena distributed in China were selected as the research objects. Four commonly used DNA barcodes(ITS2, matK, rbcL and psbA-trnH), and four highly variable regions(trnP-psaJ, psbK-psbI, trnT-trnL, clpP) in chloroplast genome were used to evaluate the identification efficiency of Dracaena species. The results showed that clpP sequence fragment could accurately identify seven species of Dracaena plants. However, due to the long sequence of clpP fragment, there were potential problems in the practical application process. We found that the combined fragment "psbK-psbI+ trnP-psaJ" can also be used for accurate molecular identification of the Resina Draconis origin plants and relative species of Dracaena, which were both relatively short sequences in the combined fragment, showing high success rates of amplification and sequencing. Therefore, the "psbK-psbI+ trnP-psaJ" combined fragment can be used as the DNA barcode fragments for molecular identification of Resina Dracon's origin plants and relative species of Dracaena. Research on the identification of Dracaena species, the results of this study can be used to accurately identify the original material of Resina Draconis, and providing effective means for identification, rational development and application of Resina Draconis base source.
China
;
DNA Barcoding, Taxonomic
;
DNA, Plant/genetics*
;
Dracaena/genetics*
;
Plants
;
Resins, Plant
;
Sequence Analysis, DNA
6.DNA Barcode Technology and Its Application Prospects in Forensic Medicine.
Jian WU ; Hong Yan GAO ; Li LUO ; Shi Tao WEN ; Peng Yu CHEN ; Jian YU
Journal of Forensic Medicine 2020;36(4):559-564
Traditional species identification has gone through five stages -- morphology, cytology, biochemistry, immunology and molecular biology. At present, the use of DNA technology for species identification has become a research hotspot. In the use of DNA for species identification, the presentation and application of DNA barcode is of epoch-making significance. With the successful application of new technology in species identification, forensic species identification has also made corresponding development, and is expected to play an important role in forensic related fields. This paper briefly describes the general situation and principles of DNA barcode technology as well as its advantages and limitations when applied to biological classification, and discusses the future significance and feasibility of DNA barcode technology in forensic applications, in order to provide new ideas for future forensic identification.
DNA/genetics*
;
DNA Barcoding, Taxonomic
;
Forensic Medicine
7.Research progress in molecular biology of Lonicerae Japonicae Flos.
Zhi-Chen CAI ; Xun-Hong LIU ; Cheng-Cheng WANG ; Meng-Xia TAN ; Jia-Li CHEN ; Yu-Qi MEI ; Li-Fang WEI ; Huan CHEN ; Rong YANG ; Jia-Jia CHEN
China Journal of Chinese Materia Medica 2020;45(6):1272-1278
Molecular biology is a new subject that clarifies the phenomena and nature of life at the molecular level. Its development provides new biotechnology and methods for the study of traditional pharmacognosy. The formation of molecular biology has brought the development of pharmacognosy into a new era of gene research. Lonicerae Japonicae Flos is a classical Chinese medicine. Many scholars of home and abroad have carried out relevant studies on its molecular biology on the basis of the in-depth study with traditional methods, and have achieved certain results. In order to provide references on the method, technical for promoting the modernization of Lonicerae Japonicae Flos, and the development, protection, and utilization of other traditional Chinese medicine resources. This article summarized the application status of molecular biology methods and techniques on the identification, biosynthesis of active constituents, and molecular mechanism of secondary metabolite under stress conditions of Lonicerae Japonicae Flos in recent years. In hybridization technology of tag(RFLP), molecular markers based on PCR(RAPD, AFLP, SSR and ISSR), based on DNA sequence analysis of SNP and DNA barcode for the variety identification, diagnosis, identification of Lonicerae Japonicae Flos, and so forth in detail. At the same time, it is proposed that multi-omics technology can be used to build systems biology technology and platforms, and establish related models of secondary metabolite biosynthesis, so as to deepen acknowledge the molecular mechanism of the active component biosynthesis of Lonicerae Japonicae Flos and the accumulation of metabolites, life activities of other medicinal plants under adverse environment, then to regulate them.
Amplified Fragment Length Polymorphism Analysis
;
Chromatography, High Pressure Liquid
;
DNA Barcoding, Taxonomic
;
Drugs, Chinese Herbal/pharmacology*
;
Lonicera/chemistry*
;
Medicine, Chinese Traditional
;
Microsatellite Repeats
;
Plants, Medicinal/chemistry*
;
Polymorphism, Restriction Fragment Length
;
Polymorphism, Single Nucleotide
;
Random Amplified Polymorphic DNA Technique
;
Secondary Metabolism
8.Identification of 23 unknown Li minority medicinal plants based on DNA barcoding.
Xin-Yun CUI ; Wei SUN ; Chao XIONG ; Xiang-Xiao MENG ; Yu-Hua SHI ; Lan WU ; Li-Li CHENG ; Wei-Jie LI ; Xi-Long ZHENG
China Journal of Chinese Materia Medica 2019;44(2):283-292
DNA barcode molecular biological technique is used to identify the species of 23 unknown Li minority medicinal plants.DNA was extracted from 23 unknown medicines using the Plant Genomic DNA Extraction kit. The ITS2 and psbA-trnH regions were amplified and sequenced bi-directionally. The Codon Code Aligner V 7. 0. 1 was used to proofread and assemble the contigs and generated consensus sequences. All the sequences were submitted to Traditional Chinese Medicine DNA Barcode Database and NCBI Gen Bank to get information of the species identifications. If the maximum similarity of the identification result is ≥ 97%,exact species can be known. If it is between 97% and 90%,samples' genus can be confirmed; If it is <90%,then we can only confirm its family. Finally there are 17 samples can be identified to species level,5 can be identified to genus level and 1 can be identified to family level. This shows that DNA barcoding used in medicinal plants molecular identification,can identify unknown species rapidly and accurately.
DNA Barcoding, Taxonomic
;
DNA, Plant
;
genetics
;
Medicine, Chinese Traditional
;
Plants, Medicinal
;
classification
;
Sequence Analysis, DNA
9.Application of DNA metabarcoding technology in identification of Chinese patent medicines.
Yu-Zhen GAO ; Jin WEI ; Zhen-Wen LIU ; Jing ZHOU
China Journal of Chinese Materia Medica 2019;44(2):261-264
Metabarcoding technology is a research method derived from the combination of traditional DNA barcodes and highthroughput sequencing technologies. It can quickly,easily and efficiently identify and restore biological samples from multiple species.Biological species are currently widely used in environmental biology research. In the market of traditional Chinese medicines,adulteration and quality instability have severely restricted the sustainable development of the related industries. This article introduced the background of the metabarcoding technology and its preliminary application in the identification of Chinese patent medicines. It also outlined the possible problems in the research process and prospected to the development of the DNA metabarcoding technology.
DNA
;
DNA Barcoding, Taxonomic
;
Drugs, Chinese Herbal
;
standards
;
Medicine, Chinese Traditional
;
Nonprescription Drugs
;
standards
10.Identification of Chrysanthemum indicum in different geographical populations and Ch. morifolium based on DNA barcodes of psbA-trnH,matK and trnL.
Fu-Rong CHEN ; Tao WANG ; Qiao-Sheng GUO ; Zai-Biao ZHU ; Feng YANG ; Qin-Jun ZOU ; Ya-Jing ZHANG
China Journal of Chinese Materia Medica 2019;44(4):660-665
DNA barcode technology was used to establish a rapid identification method of Chrysanthemum indicum and Ch. morifolium based on psbA-trn H,mat K and trn L sequences. The total DNA was extracted from 21 samples collected,and the psbA-trn H,mat K,trn L sequences were amplified by PCR and sequenced. The information of these sequences were obtained. We aligned all 63 sequences,calculated the intraspecific and interspecific distances,analysed the SNPs distribution of psbA-trn H+mat K+trn L combination sequences and constructed the Neighbor-joining( NJ) Tree,using MEGA 7. 0. The results showed that the genetic distances of Ch. indicum,Ch. indicum( Juhuanao)and Ch. morifolium were overlapped. The SNPs analysis of psbA-trn H+mat K+trn L combination sequences showed that there were 19 nucleotide polymorphism loci( SNPs) and nine parsim-informative sites in the combination sequences. In addition,Ch. indicum showed more obvious sequence polymorphism than those of Ch. indicum( Juhuanao) and Ch. morifolium. The psbA-trn H sequences showed obvious length variation.The NJ Tree showed that Ch. morifolium numbered C2-C5 were clustered into a single subbranch with a bootstrap value of 62%,and Ch.morifolium could be distinguished from Ch. indicum and Ch. indicum( Juhuanao). Moreover,Ch. indicum numbered Z9 and Z10 collected from Gansu province were singly clustered into one branch with a bootstrap value of 77%. It was also found that the changes of psbA-trn H and trn L sequences information of Ch. indicum samples from the northwest were obviously related to the geography and environment. Moreover,Ch.indicum and Ch. indicum( Juhuanao) had obvious differentiation,were also regarded as the evolutionary sources of Ch. morifolium. Therefore,psbA-trn H+mat K+trn L combination sequences as DNA barcode can identify Ch. indicum and Ch. morifolium accurately and rapidly,which provides an important basis for germplasm resources identification and species identification.
Chrysanthemum
;
DNA Barcoding, Taxonomic
;
DNA, Plant
;
Phylogeny
;
Trees

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