In order to identify Peucedani Radix, Peucedani Decursivi Radix and their adulterants, the internal transcribed spacer 2 (ITS2) regions of Peucedani Radix, Peucedani Decursivi Radix and their adulterants were amplified and bidirectionally sequenced based on the Principles for Molecular Identification of Traditional Chinese Materia Medica Using DNA Barcoding, which has been promulgated by Chinese Pharmacopoeia Commission. Sequences were analyzed and assembled by Codon Code Aligner V3. 7.1. The relevant data were analyzed by MEGA 5. 0. Species identification analyses were performed by using the nearest distance methods and neighbor-joining (NJ) methods. The result showed that the ITS2 sequence lengths of Peucedani Radix were 229-230 bp and the average intra-specific genetic distances were 0.005. The ITS2 sequence lengths of Peucedani Decursivi Radix were 227 bp and the sequences contained no variation site. The average inter-specific K2P genetic distance of Peucedani Radix, Peucedani Decursivi Radix and their adulterants species were 0.044 and 0.065 respectively. The minimum inter-specific divergence is larger than the maximum intra-specific divergence of Peucedani Decursivi Radix. The nearest distance methods and NJ trees results indicated that Peucedani Radix, Peucedani Decursivi Radix and their adulterants species could be identification clearly. The ITS2 regions can stably and accurately distinguish Peucedani Radix, Peucedani Decursivi Radix and their adulterants.
Apiaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; Drug Contamination

OBJECTIVETo study the genetic diversity of Changium smyrnioides and give a reference for utilization of the germplasm.

METHODTen different populations of Ch. smyrnioides were analyzed by the approach of sequence-related amplified polymorphism (SRAP). Genetic similarity coefficient was calculated, and systematic relationships were constructed based on the UPGMA method.

RESULTSeventeen primer pairs were selected from 160. A total of 363 bands were scored, 314 bands of them were polymorphic and the average was 18.47 polymorphic bands per primer pair, which were up to 86.50% polymorphic ratio. The results indicated that there was abundant genetic diversity among the tested materials. Genetic similarity coefficient was ranged from 0.4959 to 0.8182. Cluster analysis showed that ten different populations of Ch. smyrnioides could be distinguished into two groups.

CONCLUSIONHigh level genetic diversity was in different populations of Ch. smyrnioides, and genetic relationship was correlative to geographic position.

Apiaceae ; classification ; genetics ; Genetic Variation ; genetics ; Nucleic Acid Amplification Techniques ; methods ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction

In this study, 31 Notopterygium incisum populations were analyzed using ITS sequences to investigate the genetic structure. The results showed that: the ITS region ranged in size from 634 to 635 bp and base composition was with high G + C content of 57.8%. Thirty-one polymorphic sites were detected from 402 sequences of 31 populations of N. incisum, and the proportion of polymorphic sites was 4.88%, in which parsimony informative sites were up to 12. And 31 haplotypes were identified based on these polymorphic sites. Molecular variance analysis (AMOVA) indicated that high genetic differentiation (57%) existed among population, and gene flow was low (N(m) = 0.38) among populations. Phylogenetic relationships of 31 haplotypes were analyzed using NJ method with N. forbesiias an out-group. Phylogenetic analysis showed that 31 haplotypes from different populations mixed together and did not form distinct geographically separated clades.
Apiaceae ; classification ; genetics ; Base Sequence ; China ; DNA, Intergenic ; genetics ; Gene Flow ; Genetic Variation ; Molecular Sequence Data ; Phylogeny

In this study, Notopterygii Rhizoma et Radix was used to verify the stability and accuracy of DNA barcodes in identification of Chinese materia medica for the first time. All genomic DNAs from thirty one samples were extracted. The ITS (internal transcribed spacer) regions were amplified and sequenced bi-directionally. Obtained sequences were assembled using the CodonCode Aligner. And the sequences of the ITS regions were aligned through Clustal-W and the genetic distances were computed using MEGA 5.0 in accordance with the kimura 2-parameter (K2P) model. The neighbor-joining (NJ) phylogenetic trees were constructed. The ITS2 regions were obtained by using the hidden Markov model (HMM)-based annotation methods from the ITS sequences. Results indicated that the lengths of ITS regions of Notopterygii Rhizoma et Radix were 603-604 bp, while the lengths of ITS2 regions were 228 bp. The haplotypes of ITS/ITS2 regions of Notopterygii Rhizoma et Radix were the same as those of the original plant leaves. The intra-specific genetic distances were smaller than inter-specific ones in ITS/ITS2 regions of Notopterygium incisum and N. franchetii. The NJ trees showed that N. incisum, N. franchetii and its adulterants can be easily differentiated according to their monophyly. Therefore, ITS/ITS2 regions as DNA barcodes can stably and accurately distinguish Notopterygii Rhizoma et Radix from its adulterants and could provide a new technique to ensure clinical safety in utilization of traditional Chinese medicines.
Apiaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Phylogeny ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; Rhizome ; genetics

OBJECTIVETo explain the molecular evidence of revision of taxonomic placement of Peucedanum decursivum based on the nrDNA ITS sequence.

METHODPCR amplification, DNA sequencing and cladistic analysis.

RESULTThe ITS sequences and phylogenetic tree of 5 species of Angelica were and Peucedanum were acquired, in which 5 species were divided into 2 groups, Angelica group and Peucedanum group. P. decursivum was placed in the Angelica group.

CONCLUSIONP. decursivum belongs to genus Angelica. The scientific name of P. decursivum should be revised as A. decursivum. A. decursivum and P. praeruptorum should be used as crude drug respectively.

Angelica ; classification ; genetics ; Apiaceae ; classification ; genetics ; Base Sequence ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Leaves ; genetics ; Plants, Medicinal ; classification ; genetics

Saposhnikovia divaricata is a valuable Chinese medicinal herb; the transformation from vegetative growth to reproductive growth may lead to the decrease of its pharmacological activities. Therefore, the study of bolting and flowering for Saposhnikovia divaricata is warranted. The present study aimed to reveal differentially expressed genes (DEGs) and regularity of expression during the bolting and flowering process, and the results of this study might provide a theoretical foundation for the suppression of early bolting for future research and practical application. Three sample groups, early flowering, flower bud differentiation, and late flowering (groups A, B, and C, respectively) were selected. Transcriptomic analysis identified 67, 010 annotated unigenes, among which 50, 165 were differentially expressed including 16, 108 in A vs B, and 17, 459 in B vs C, respectively. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway functional classification analysis were performed on these differentially expressed genes, and five important pathways were significantly impacted (P ≤ 0.01): plant circadian rhythm, other glycan degradation, oxidative phosphorylation, plant hormone signal transduction, and starch and sucrose metabolism. Plant hormone signal transduction might play an important role in the bolting and flowering process. The differentially expressed indole-3-acetic acid (IAA) gene showed significant down-regulation during bolting and flowering, while the transport inhibitor response 1 (TIR1) gene showed no significant change during the bolting process. The expression of flowering related genes FLC, LYF, and AP1 also showed a greater difference at different development stages. In conclusion, we speculate that the decrease in auxin concentration is not caused by the degrading effect of TIR1 but by an alternative mechanism.
Apiaceae ; genetics ; growth & development ; Flowers ; genetics ; growth & development ; Gene Expression Profiling ; Gene Expression Regulation, Plant ; Gene Regulatory Networks ; Genes, Plant ; RNA, Plant ; genetics ; Reproducibility of Results

Saposhnikovia divaricata is a commonly used bulk medicinal plant. To explore the key enzyme genes and their expression in the biosynthesis of chromone and coumarin, the key active components, we carried out transcriptome sequencing(Illumina HiSeq) and bioinformatics analysis for the 1-year-old(S1) and 2-year-old(S2) plants of S. divaricata. A total of 40.8 Gb data was obtained. After the sequence assembly via Trinity, 110 732 transcripts and 86 233 unigenes were obtained, which were aligned and annotated with NR, Swiss-Prot, GO, KEGG, and PFAM. Daucus carota and S. divaricata had the highest sequence homology. KEGG pathway enrichment showed that the differentially expressed genes were mainly enriched in plant hormone signal transduction, phenylpropanoid biosynthesis, and flavonoid biosynthesis pathways. A total of 27 differentially expressed unigenes, including 13 enzyme genes, were identified in the pathways related to the synthesis of active ingredients in S. divaricata. Compared with S1 plant, S2 plant showed up-regulated expression of PAL, BGL, C4H, 4CL, CYP98A, CSE, REF, and CCoAOMT and down-regulated expression of CHS, CAD, and COMT. HCT and POD had both up-regulated and down-regulated unigenes. Among them, PAL, C4H, 4CL, BGL, and CHS can be used as candidate genes for the synthesis of the active ingredients in S. divaricata. The four key enzyme genes were verified by RT-qPCR, which showed the results consistent with transcriptome sequencing. This study enriches the genetic information of S. divaricata and provides support for the identification of candidate genes in the biosynthesis of secondary metabolites.
Apiaceae/genetics* ; Chromones ; Coumarins ; Flavonoids ; Gene Expression Profiling ; Gene Expression Regulation, Plant ; High-Throughput Nucleotide Sequencing/methods* ; Plant Growth Regulators ; Transcriptome

OBJECTIVETo assess the population genetic diversity and genetic structure and screen species-specific bands for identification of Changium smyrnioides and Chuanminshen violaceum.

METHODSeven wild populations of Changium smyrnioides and one cultivated population of Chuanminshen violaceum were studied by ISSR analysis. The population genetic diversity and population genetic structure were assessed by using POPGENE software.

RESULTA total of 152 ISSR markers were scored, among which 136 (90.8%) were polymorphic. The values of Gst tended to be high (mean Gst = 0.575). The level of genetic divesity of Changium smyrnioides (A = 1.272; P = 27.26%; I = 0.132; H = 0.087) was higher than that of Chuanminshen violaceum (A = 1.217; P = 21.7; I = 0.103; H = 0.067).

CONCLUSIONThe genetic variation of Changium smyrnioides is high and the majority of genetic variation occur among populations. Substantial genetic divergence is shown by cluster analysis (UPGMA) to befound between Changium smyrnioides and Chuanminshen violaceum at DNA level. In addition, one species-specific marker has been obtained in Chuanminshen violaceum. The phylogenetic relationship of two species has also been discussed.

Apiaceae ; classification ; genetics ; China ; Cluster Analysis ; DNA, Plant ; genetics ; Ecosystem ; Gene Frequency ; Genetic Markers ; Genetic Structures ; Phylogeny ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Repetitive Sequences, Nucleic Acid ; Species Specificity

This study is purposed to investigate the effects of praeruptorin A (PA) and praeruptorin C (PC) on UGT1A1 in HepG2 cells through hCAR pathway. PA and PC were incubated with HepG2 cells for 24 h and 48 h, mRNA and protein expressions of UGT1A1 were determined by real-time PCR and Western blotting assays. Additionally, effects of PA and PC on UGT1A1 mRNA and protein expressions were also measured after transient transfection of a specific CAR siRNA for 72 h in HepG2 cells. UGT1A1 mRNA and protein expression levels were significantly increased by PA and PC after incubation for 48 h. Moreover, the mRNA and protein up-regulations of UGT1A1 were attenuated by transient transfection of a specific CAR siRNA, suggesting the induction was mediated by CAR. The results suggest that PA and PC can significantly up-regulate UGT1A1 expression partially via the CAR-mediated pathway.
Apiaceae ; chemistry ; Coumarins ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Glucuronosyltransferase ; genetics ; metabolism ; Hep G2 Cells ; Humans ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; metabolism ; Receptors, Cytoplasmic and Nuclear ; genetics ; metabolism ; Signal Transduction ; Transfection

OBJECTIVETo study the effects of volatie oil of Schizonepeta tenuifolia Briq herb and Saposhnikovia divaricata Schischke root (OSS) on proinflammatory cytokine expression and regulation in rats.

METHODOA and LPS were injected intravenously to rats to develop acute lung injury (ALI). The rats were treated with OSS (45.19 microL kg(-1)). The pathological sections of lung tissue were prepared and observed in acute lung injury rats. The expression of nuclear factor-kappa B p65 (NF-kappaB p65), intercellar adhesion molecule CD54, and NF-kappaB p65 mRNA were determined in lung cells.

RESULTvolatie oil of Schizonepeta tenuifolia Briq herb and Saposhnikovia divaricata Schischke root significantly inhibited the expression of CD54, the activation of NF-kappaB p65, and the transcription of NF-kappaB p65 mRNA.

CONCLUSIONOSS can reduce the expression of CD54 and NF-kappaB p65 protein synthesis, which may be its anti-inflammatory molecular mechanisms.

Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Apiaceae ; chemistry ; Drug Combinations ; Gene Expression Regulation ; drug effects ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Lamiaceae ; chemistry ; Lipopolysaccharides ; Male ; Oils, Volatile ; isolation & purification ; pharmacology ; Oleic Acid ; Plant Oils ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Respiratory Distress Syndrome, Adult ; chemically induced ; metabolism ; prevention & control ; Transcription Factor RelA ; biosynthesis ; genetics