RESUMO
Objective System evolution relationship and molecular identification method of the germplasm resources of Lycoris aurea from different regions was analyzed based on the sequence of psbA-trnH chloroplast gene. Methods DNA samples of 52 L. aurea populations were extracted from 15 provinces or cities in China. The psbA-trnH sequences of the populations were amplified by PCR, and the purified PCR products were sequenced and analyzed by Mega 5.0 software etc. Results The length of psbA-trnH sequences were 544-656 bp, and GC content of them was 35.8%-37.0%, and the genetic distances among the populations were 0-0.009 47. There were 33 variable (polymorphic) sites, including nine parsimony informative sites and 18 singleton variable sites and six insertion/deletion gaps. Ten haplotypes (H) were identified. Values of haplotype diversity (Hd) and nucleotide diversity (π) were 0.749 and 0.002 63, respectively. The genetic diversity of the populations of L. aurea were very high. In the maximum parsimony phylogenetic tree, 52 populations of L. aurea were clustered into four branches, which was almost consistent with their geographical distributions. Conclusion The genetic variation of L. aurea populations from different regions is significant and the psbA-trnH sequence could be used as a molecular evidence for identifying the germplasm resources of L. aurea from different regions. There is very obvious regional characteristics in evolution for germplasm resources of L. aurea in China.
RESUMO
Objective: To explore the optimum condition for complex enzyme-assisted extraction of galanthamine from Lycoris aurea by L9 (34) orthogonal array design and separation effect of cation exchange resin on galanthamine. Methods: Cellulase and pectinase solution was used as the extraction solvent. The effects of pH value of enzyme, amount of complex enzyme, dissociation time, and enzymatic hydrolysis temperature on the extraction results were investigated. Results: The optimal conditions were obtained as follows: ratio of solid to liquid (g: mL) 1:10, pH value 4.5, amount of complex enzymes 4%, enzymatic hydrolysis temperature 50 °C, and reaction time 2.0 h. Under these conditions, the extraction yield of galanthamine was 0.0294%. In addition, D-001 cation exchange resin was selected for separation of galanthamine. The separation conditions were that adsorption flow rate was 3 BV/h with reagent of pH 2 and the desorption flow rate was 3 BV/h with 70% ethanol solution containing 1.5 mol/L ammonia. After separation, the content of galanthamine was increased to 12.31%. Conclusion: The results provide a reference for industrial production of galanthamine.
RESUMO
In this study, we investigated the effect of methanolic extract isolated from the root of Lycoris aurea (LA) on the growth of cancer cells and the tube formation activity of endothelial cells. Various cancer cells were treated with LA at doses of 0.3, 1, 3, 10 or 30 microg/ml and LA significantly suppressed the growth of several cancer cell lines, including ACHN, HCT-15, K-562, MCF-7, PC-3 and SK-OV-3, in a dose-dependent manner. We also found that LA induced cell cycle arrest at G2/M phase in ACHN renal cell adenocarcinoma cells. Further study demonstrated that LA concentration-dependently inhibited the tube formation, which is a widely used in vitro model of reorganization stage of angiogenesis, in human umbilical vein endothelial cells. Collectively, these results show that LA inhibits the growth of cancer cells and tube formation of endothelial cells and the growth-inhibitory effect of LA might be mediated, at least in part, by blocking cell cycle progression.