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Objective To obtain the transcriptome sequence database and to identify the genes related to the biosynthesis of anthraquinones in Rubia cordifolia. Methods The transcriptome data of root of Rubia cordifolia was obtained by Illunima HiseqTM 2000 150PE sequencing and de novo splicing of Unigene was realized by Trinity. The GO classification, KOG functional, metabolism of KEGG metabolic pathway and protein function annotation analysis were completed based on BLAST. Results 7.3 Gb database was assembled after assembly steps, and 66 646 unigenes were assembled with an average length of 1 424 bp. All Unigenes were annotated in the public databases NR (NCBI nonredundant protein sequence), NT (NCBI nucleotide sequences), KEGG (kyoto encyclopedia of genes and genomes), Swissprot (A manually annotated and reviewed protein sequence database), GO (gene ontology), KOG (euKaryotic ortholog groups) and Pfam (protein family). Based on the assignment of KEGG pathway, 64 Unigenes involved in anthraquinones biosynthesis were found. Conclusion This study was the first comprehensive transcriptome analysis for R. cordifolia, and the candidate genes involved in the biosynthesis of anthraquinones were obtained. The transcriptome data constitutes a much more abundant genetic resource that can be utilized to benefit further molecular biology studies on R. cordifolia.
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Rubia cordifolia L. has been used as a traditional Chinese medicine for several centuries. In recent years, the resources of wild Rubia cordifolia have been declined sharply due to increased utilization and rising price. Therefore, it is of great urgency to evaluate and protect resources of wild plant of Rubia cordifolia. In our study, sixty-four individuals that represent eight wild populations of Rubia cordifolia L. were analyzed by Start Codon Targeted Polymorphism (SCoT) molecular markers. Genetic distance was calculated by POPGENE 3.2 software, cluster analysis was generated by NTSYS 2.10 software based on UPGMA method and Mantel Test was used to analysis the relationship between the genetic distances and geographical distance among the wild populations. The results showed a high genetic diversity of wild populations of Rubia cordifolia L. in Shaanxi province. A total of 182 bands were produced by 14 primers, among which 163 bands were polymorphic bands, and the percentage of polymorphic bands was 89.56%. The average value of Nei's genetic diversity index (H) was 0.293 6, Shannon's information index (I) was 0.444 6, genetic differentiation coefficient (Gst) was 0.555 3, and the gene flow (Nm) was 0.440 8, the wild populations were ranked by genetic diversity:AK > YL > SL > BJ > TC > YA > WN > XY. Mantel Test analysis demonstrated that the significant correlation was found between the genetic distances and geographical distances (r=0.776 4, PRubia cordifolia L. germplasms.
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Objective To evaluate the effects of a traditional Chinese medicine Rubia cordifolia L.aqueous extract (RCAE) on body weight, fat mass and parameters of glucose and lipid metabolism in high fat diet (HFD)-induced obese rats and its mechanism.Methods pGL3-Enhancer-PPARγ2 (625 bp)-Luc plasmid, a luciferase reportergene expression plasmid containing PPARγ2 promoter was constructed and stably transfected 3T3-L1 preadipocytes were established.PPARγ2 promoter`s activities in these cells were detected after administration with different concentration (0.1 mg/L~1 000 mg/L) of RCAE or with 100 mg/L RCAE for different action time.PPARγ2 mRNA expression in human adipocytes were detected after administration with 100 mg/L RCAE.Meanwhile, HFD-induced obese rats were administrated with low or high dose RCAE to investigate the effects of RCAE on serum glucose, lipid and insulin levels, body weight, visceral fat mass and so on.Results 10 mg/L RCAE could increase luciferase expression in 3T3-L1 cells to 1.43 folds of that in control group (P<0.01) and it reached 3.24 folds of that in control group when the concentration of RCAE was 1000 mg/L (P<0.01).With the administration with 100 mg/L RCAE, the luciferase activity of 3T3-L1 cells peaked at 28 h where it was 2.72 folds of that in control group (P<0.01), and the expression of PPARγ2 mRNA in human adipocytes increased to 2.27 folds of that in control group (P<0.01).Compared with HFD group, low dose RCAE significantly reduced the fasting insulin level, HOMA-IR and visceral fat mass (P<0.05).Conclusions Low dose RCAE significantly reduces the visceral fat mass and ameliorates insulin resistance in HFD-induced obese rats.The potential mechanism may be explained by the stimulation of PPARγ2 promoter activities and the increased expression of PPARγ2 gene.
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Objective To investigate the chemical constituents of the aerial part of Rubia cordifolia. Methods All compounds were isolated and purified by silica gel, Sephadex LH-20, and MCI column chromatography. Their structures were determined by physicochemical properties and spectral data. Results Eleven compounds were isolated and identified as: (-)-episyringaresinol (1), (-)-secoisolariciresinol (2), (-)-3,4-divanillyl tetrahydrofuran (3), lignans (+)-demethoxypinoresinol (4), (+)-syringaresinol (5), (+)-isolariciresinol (6), burselignan (7), (7S,8R)-dihydrodehydroconiferyl alcohol (8), 4,5'-dimethoxy-lariciresinol (9), (+)-7R,8S-5- methoxydihydrodehydroconifery alcohol (10), and 5,5'-dimethoxy-7-oxolariciresinol (11). Conclusion All compounds are isolated from this plant for the first time.
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Objective: To investigate the genetic diversity of Rubia cordifolia from seven geographical populations in Henan province, and to provide evidence for the protection of R. cordifolia wild resources. Methods: The autogenous variation of chloroplast gene fragment psbA-trnH was analyzed by DNA sequencing technique. Results: Based on cpDNA data, 40 haplotypes (H1-H40) were identified, the diversity index of haplotype and nucleotide diversity index were 0.951 ± 0.0013 and 0.01477, respectively. Genetic differentiation index was 0.08153, and gene flow among populations was 2.78. Conclusion: The wild populations of R. cordifolia in Henan province have higher genetic diversity in geo-authentic product area than other species. There is less genetic differentiation among populations because of the stronger seed-mediated gene flow.