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ObjectiveTo conduct phylogenetic analysis of internal transcribed spacer 2 （ITS2） and chloroplast gene segments including psbA-trnH， rbcL， and matK of Sophora japonica cv. jinhuai resource samples from different geographical sources， and to explore the genetic diversity of S. japonica cv. jinhuai. MethodPolymerase chain reaction （PCR） method was used to amplify the nucleic acid sequences of ITS2， psbA-trnH， rbcL， and matK of S. japonica cv. jinhuai. Neighbor joining （NJ） method was used to construct phylogenetic trees， and Kimura 2-Parameter （K2P） model was used to calculate the genetic distance of different samples. MEGA and BIOEDIT softwares were applied for mutiple alignment and analysis of ITS2， psbA-trnH， rbcL， and matK sequences of S. japonica cv. jinhuai. ResultThe lengths of ITS2 sequence were 278-279 bp. The lengths of psbA-trnH were 289 bp. The lengths of rbcL sequence were 673 bp. The lengths of matK sequences were 786-792 bp. There were 3 mutation points in ITS2 and psbA-trnH， no mutation point in rbcL， and 13 mutation points in matK. The samples of S. japonica cv. jinhuai were clustered into two groups based on the phylogenetic tree constructed by ITS2 sequences. The sample of seedling tree in Baibao was clustered into one group， while the other 25 samples were clustered into another group. For the psbA-trnH sequence， the success rate of PCR amplification of 28 samples of S. japonica cv. jinhuai was 100%. The 28 samples of S. japonica cv. jinhuai were clustered into three groups based on the clustering results of psbA-trnH sequence. The sample of seedling tree in Shaoshui was clustered into one group. The five samples of grafting tree and seedling tree in Miaotou， grafting trees in Jiantang， Wenqiao， and Daxu， and seeding tree in Xianshui were clustered into one group. The other 21 samples were clustered into another group. The 26 samples of S. japonica cv. jinhuai were clustered into two groups based on the phylogenetic tree constructed by matK sequences. The sample of seedling tree in Xianshui was clustered into one group， while the other 25 samples were clustered into another group. The clustering results of the rbcL sequence of S. japonica cv. jinhuai could not distinguish 28 resource samples. The phylogenetic tree constructed by the combined sequence of ITS2+psbA-trnH+rbcL+matK divided S. japonica cv. jinhuai resource samples into 4 groups. The 13 samples of seedling trees in Qiyang， Daoxian， Miaotou， Shaoshui， Shitang， Xianshui， Jiantang， and Xiangli， and grafting trees in Qiyang， Miaotou， Yongsui， Wenqiao， and Yangtang were clustered into one group. The sample of seedling tree in Wenqiao was clustered into one group. The sample of seedling tree in Daxu was clustered into one group. The remaining samples were clustered into another group. ConclusionPhylogenetic and mutation analysis provide the theoretic foundation to investigate the evolution of the resources of S. japonica cv. jinhuai， and evaluate their genuineness. The results of mutation points can be used to identify the related S. japonica cv. jinhuai resources. The findings of this study show that the combination of different gene sequences has an optimal effect on plant identification.
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Objective: To provide a basis for the rational use of Sophora japonica resources through comprehensive evaluation of different tissues and organs. Methods: The contents of rutin, narcissin, quercetin, isorhamnetin and heavy metals in the samples were detected by HPLC and ICP-OES. The Fe3+ reducing ability, DPPH free radical (DPPH•), ABTS free radical (ABTS•+) scavenging ability and total antioxidant capacity (FRAP) were detected by colorimetry. Then, cluster analysis (CA) and principal component analysis (PCA) were conducted by software of SPSS 20.0. Results: The total contents of four flavonoids in different tissues and organs of S. japonica were arranged as follows: flower buds > flowers > flower axis > leaves > branches. The order of antioxidant capacities was as follows: flower buds > flowers > flower axis > leaves > branches, which were positively correlated with the total contents of four flavonoids. The contents of five heavy metal elements in flowers and flower buds were within the limitation of the Green standards of medicinal plants and preparations for foreign trade and economy, while the Cd element in some leaves, flower axis and branches was beyond the standard. Flowers and flower buds were clustered into one type by CA, while flower axis, leaves and branches were clustered into another category. The two principal components (PC1 and PC2) were extracted from the eight variables by PCA, PC1 showed significant differences among different tissues and organs, and PC2 values showed large differences among batches. Conclusion: The flowers and flower buds of S. japonica showed an excellent qualities, including safe doses of heavy metals, rich flavonoids and outstanding antioxidant activities. In addition, the flower axis and leaves also contained high flavonoids and exhibited strong antioxidant activities, which had the value of further development and utilization.
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In order to study the pathways of biosynthesis of flavonoids in Sophora japonica, 113 797 unigenes were obtained by Trinity software, with an average length of 803 bp, of which 72 752 unigenes were obtained from the database by high-throughput sequencing, and a total of 38 891 SSR loci were searched. Through the metabolic pathway analysis, we found that there were 135 unigenes involved in the biosynthesis of flavonoids and 959 unigene involved in other secondary metabolic pathways. Further analysis of genes involved in rutin biosynthesis revealed that 24 were associated with CHS, 52 were associated with FLS, and 11 were associated with UFGT. The obtained data of S. japonica transcriptome lays the foundation for studying the pathways of biosynthesis of flavonoids in S. japonica and provides theoretical basis for the formation of the quality of S. japonica.
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This work aimed to study the antioxidant activity of a quercetin-containing flavonoid extract (FQЕ) obtained from Sophora japonica L. flower buds rich in quercetin (91.6%). Radical scavenging activity was analyzed towards the synthetic radicals DPPH and ABTS+ and antioxidant activity was evaluated applying the method of oxygen consumption in a model system containing methyl linoleate. Model food systems of lard and sunflower oil were explored by the application of Rancimat method and chicken as a real food system was investigated by the thiobarbituric acid test. Results showed a high radical scavenging activity and antioxidant capacity of QFE similar to those of the pure flavonoid quercetin.
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Objective: To establish the callus culturing system of embryos in Sophora japonica, and to analyze the contents of isoflavones in callus. Methods: Through adding the plant growth regulators of different kinds and at different concentration, the optimal media for callus induction, suspension cell culture, and the highest isoflavone content were obtained. Results: The optimal medium for callus induction of embryos in S. japonica was B5 + 2, 4-D (1.0 mg/L) + 6-BA (0.2 mg/L) + sucrose (20 mg/L). In solid medium, the callus was growing rapidly after 7 d, and the yield reached to the highest on the day 35 (fresh weight 7.4137 g), but the isoflavone content had no significant changes. In suspension culture, the yield of callus reached to the highest on the day 24 (fresh weight 11.563 8 g), and the callus turned into decline phase after 24 d, in which the fresh and dry callus weights were descending. With continuous culture, the cells were browning gradually, aging, and dying at last, but the isoflavone content was opposite to growth curve, and reached its lowest point (4.826 mg/g) on the day 24. Conclusion: The induction and cultivation of embryos callus in S. japonica and the isoflavone content have a larger correlation with the plant growth regulators of the kinds and the concentration.
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Objective: To compare the chromatographic profiles and contents of three flavonoid aglycone equivalents (quercetin, genistein, and kaempferol) in the fruits of Sophora japonica var. pendula Loud. and the authentic Fructus sophorae, so as to provide a theoretical basis for using the fruit of Sophora japonica var. pendula Loud. as a substitute for Fructus sophorae. Methods: The fruits of Sophora japonica var. pendula Loud. were collected from different areas of Shijiazhuang in Winter. The flavonoids in the fruits were extracted by successive extraction with methanol. The acidic-hydrolyzed extracts were separated on a Diamonsil C18 column (150 mmX4.6 mm, 5μm). 0.4% phosphoric acid-methanol (50 ; 50, V/V) was used as mobile phase. The flow rate of mobile phase was 1.0 ml/min and the effluents were monitored at 360 nm. Results: The chromatographic profiles of the two fruit extracts were consistent basically. The contents of quercetin, genistein, and kaempferol in the fruits of Sophora japonica var. pendula Loud. were (0.30±0.07) %, (3.04±0.27) %, and (2.74±0.30)%, respectively, which were similar to those in the authentic Fructus sophorae. Conclusion: It is found that the contents of major flavonoid aglycones in the fruit of Sophora japonica var. pendula Loud. are similar to those in the authentic Fructus sophorae, making it a potential substitute for the Fructus sophorae.
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OBJECTIVE:To establish the quality standards of processed Sophora japonica.METHODS:Different batches of processed S. japonica products were detected in respect of property,microstructure,TLC,moisture,total ash,acid insoluble ash,extract inspection,content of total flavonoids and rutin. RESULTS:The limit value of test term for S. japonica,stir-baked S. japonica,S. japonica carbon were confirmed. CONCLUSION:Established standard is applicable for the quality control of processed S. japonica.
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OBJECTIVE: To optimize the technology of extracting total flavonoids from Sophora japonica.METHODS: The extracting technology of total flavonoids (from Sophora japonica) was optimized by response surface methodology(RSM) with ethanol concentration,solid-solvent ratio,extracting temperature,extracting time and extracting times as factors and the yield of total flavonoids as index.RESULTS: The optimal parameters were determined as follows: the ethanol concentration was 60%;the solid-solvent ratio was 1∶10;the extracting was conducted for 1 time(90 min) at 90 ℃.CONCLUSION:The optimized extraction technology was simple and stable and which serves as references for industrial large scale production.
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Flowers of Sophora japonica were processed at nature conditions in Ho Chi Minh City after followed the stability within 2 years by rutin quantitative method follow Vietnamese pharmacopoeia III- 2002 by spectrophotometer and quantify quercetin by High-Perfomance Liquid Chromatography (HPLC). Results: the content of rutin in flowers of Sophora japonica decreased across preservation time (from 6 to 11%) and quercetin mixture did not increase significantly (approximate 0.1%). The rutin content of flowers of Sophora japonica was still high compared to Vietnamese pharmacopoeia III- 2002 standards. Thus, the shelf-life of the flowers of Sophora japonica is proposed to be 30 months if well preserved
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Flowers , Drug StabilityABSTRACT
Object To select the optimum extracting procedure for quercetin from bud of Sophora japonica L. Methods The optimum extracting procedure was selected with the content and extraction effeciency of quercetin from bud of S. japonica by orthogonal test design. Results The optimum extracting procedure was determined with 10 times of 0.05% sodium hydroxide decocting 20 min and 4 times, adjusting pH value to 6, hydrolyzing with 10 times of 4% sulfuric acid for 1 h after precipitated. Conclusion The optimum extracting procedure increased the content and extraction efficiency of quercetin from bud of S. japonica.
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AIM: To evaluate the effect of microfiltrating the extract of Sophora japonica L. by inorganic ceramic membrane. METHODS: The extract of Sophora japonica L. was processed by inorganic ceramic membrane, and the whole solid, effective ingredinent and flux of mebrane were examined. RESULTS: The extract of Sophora japonica L. became clear after microfiltration. The decreasing rate of the whole solid was 27.88%, the metastasis rate of the effective ingredient was 78.26, and the increasing rate of the effective ingredient was 8.48%. CONCLUSION: The microfiltration of inorganic ceramic membrane can improve the qualitry of extract of Sophora japonica L. refinery effect.