Human pulp tissue dissolution ability of different extracts of Sapindus mukorossi: An in vitro study / 中草药·英文版

Objective: Due to the many negative properties of sodium hypochlorite used in current root canal treatment, interest in biocompatible natural agents is increasing day by day. The aim of this study was to evaluate whether various extract solutions of Sapindus mukorossi have dissolution effects on human pulp tissues. Methods: Primarily powder extracts were obtained by extracting fruit shells of S. mukorossi in different solvents (ethanol, methanol, buthanol and distilled water). The test solutions were prepared and randomly separated into six groups with 10 samples in each group: ethanol extract, methanol extract, butanol extract, distilled water extract of S. mukorossi, sodium hypochlorite (NaOCl) and the control group. Among these, S. mukorossi extracts were separated into two subgroups, depending on their concentration level (50 µg/mL and 100 µg/mL). The pulp tissues of freshly extracted human molars were used for dissolution test. The weights of the pulpal tissues were measured and recorded for two times after the samples were placed in the solutions. Statistical analysis for all descriptive statistics was performed using SPSS 22 (P < 0.05). Results: Our results showed that maximum percent yield of preparation was obtained in methanol extract of S. mukorossi. Among all of the groups, the best dissolution capacity was seen in the NaOCl group (positive control group). Among S. mukorossi groups, the best tissue solvent solution was found in SMM group at 50 µg/mL and SMB group at 100 µg/mL. Conclusion: The different extracts of S. mukorossi had a capacity to dissolve pulp tissue but this capacity was less than NaOCl. Therefore, further studies will enable the creation of a commercial solution for clinical use by increasing the effectiveness of S. mukorossi while combining it with other endodontic irrigation solutions.

Chemical constituents from stems and leaves of mangrove plant, Scyphiphora hydrophyllacea / 中草药

Objective: To study the chemical constituents of the stems and leaves of mangrove plant, Scyphiphora hydrophyllacea. Methods: The chemical constituents of S. hydrophyllacea were separated and purified by silica gel, ODS, Sephadex LH-20 gel column chromatographies and preparative HPLC. Their structures were identified by physicochemical properties, spectroscopic analysis, as well as comparisons with the data reported in literatures. Results: A total of 18 compounds were isolated from the 90% ethanol extract of the stems and leaves of S. hydrophyllacea, which were identified as pomonic acid (1), euscaphic acid (2), dammaradienyl acetate (3), erythrodiol (4), lupenyl acetate (5), 30-oxo-lupeol (6), maslinic acid (7), 23-hydroxyursolic acid (8), β-amyrenone (9), acacetin (10), chrysoeriol (11), jaceosidin (12), kumatakenin (13), isosakuranetin (14), taxifolin (15), lyoniresinol (16), medioresinol (17), and balanophonin (18). Compounds 1-9 were triterpenoids, compounds 10-15 were flavonoids and compounds 16-18 were lignans. Conclusion: All compounds are isolated from the genus Scyphiphora for the first time.

Research advances on chemical constituents and pharmacological effects of various parts of Nelumbo nucifera / 中草药

As a common medicine and homologous plant, Nelumbo nucifera mainly contains alkaloids, flavonoids, glycosides, terpenes, steroids, fatty acids, proteins, minerals, vitamins and other chemical constituents, with lipid-lowering, antibacterial, anti-inflammatory, antioxidant, hemostasis and other pharmacological activities. After reviewing the literatures at home and abroad for nearly 40 years, 385 compounds have been reported from different parts of N. nucifera including lotus leaves, plumula nelumbinis, lotus, lotus seeds, lotus root, lotus seedpod, nelumbinis rhizome node, lotus stem, N. nucifera stamens and lotus seed skins. There are 86 alkaloids, 133 flavonoids, and 166 other compounds. In this review, we summarized the chemical constituents and pharmacological effects reported in N. nucifera, and it provides a reference for further study on the chemical constituents, pharmacological activity and development and utilization of N. nucifera.

Hypoglycemic and antioxidant activities of Clerodendrum inerme leaf extract on streptozotocin-induced diabetic models in mice / 中草药·英文版

Objective: Recently, much attention has been paid to natural product-derived compounds for antidiabetic drug discovery. More recent studies are being focused on clarifying the bioactivity of plants and derived products. The aim of the present study was to investigate the anti-oxidant and antidiabetic activities of Clerodendrum inerme leaf extract (CILE) in streptozotocin-induced diabetic mice. Methods: C. inerme leaves were analyzed for preliminary phytochemical properties and the content of total phenolic and flavonoid were determined. In vitro anti-oxidant activity was measured using DPPH assay. Streptozotocin-induced diabetic model in mice was applied for in vivo study by the effect of CILE at two dose levels (343 and 686 mg/kg b.w.). Results: The results showed that C. inerme leaves contained the major constituents of flavonoids, alkaloids, tannins, triterpenes, and saponins. CILE exhibited the total polyphenol and flavonoid content with 120.458 mg gallic acid equivalent/g dry weight and 4.494 mg hispidulin equivalent/g dry weight, respectively. The anti-oxidant activity of CILE was expressed with IC50 = 25.28 µg/mL. CILE at the doses of 343 mg/kg and 686 mg/kg after 7 d administration exerted a decrease in plasma glucose, protected the liver, kidneys against oxidation stress via increasing glutathione content in the liver, and reduced malondialdehyde content in the liver and kidneys. Pancreatic histological analysis in diabetic mice treated with CILE also showed the pancreatic β-cells regeneration via increasing the size and number of pancreatic islets. Conclusion: These findings suggested that C. inerme leaves had potent antidiabetic and anti-oxidant activities. The results provide reliable scientific base, which is the premise for further research and development of CILE as supplements.

Lignans from seed of Hydnocarpus anthelminthica / 中国中药杂志

This project is to investigate lignans from the seed of Hydnocarpus anthelminthica. Thirteen lignans were isolated from the 95% ethanol extract of the seed of H. anthelminthica, by polyamide resin, Sephadex LH-20, ODS column chromatography and preparative HPLC. Their structures were elucidated as(+)-syringaresinol(1), lirioresinol A(2),(+)-medioresinol(3),(7R,8R,8'R)-4'-guaiacylglyceryl-evofolin B(4), leptolepisol C(5),(-)-(7R,7'R,7″R,8S,8'S,8″S)-4',4″-dihydroxy-3,3',3″,5,5',5″-hexamethoxy-7,9':7',9-diepoxy-4,8″-oxy-8,8'-sesquineolignan-7″,9″-diol(6),(-)-(7R,7'R,7″R,8S,8'S,8″S)-4',4″-dihydroxy-3,3',3″,5,5'-pentamethoxy-7,9':7',9-diepoxy-4,8″-oxy-8,8'ses-quineolignan-7″,9″-diol(7), ceplignan(8), hydnocarpusol(9), isohydnocarpin(10),(-)-hydnocarpin(11), hydnocarpin(12), and hydnocarpin-D(13) by spectroscopic data analysis. Compounds 1-8 were obtained from the genus Hydnocarpus for the first time.

Preliminary Phytochemical Analysis Of Leaves Of Guchakaranja (quassia Indica Gaertn)

The drug Guchakaranja is mentioned as one of Karanja variety by Raj Nighantu and Nighantu Ratnakara. The references of the drug are not seen in other classical textbooks of Ayurveda and the only references seen in these two Nighantus. The drug had been correlated to Karinjotta, a locally available plant in Kerala. The drug Guchakaranja is botanically correlated as Quassia indica Gaertn (Samadera indica Gaertn) belonging to the family Simaroubaceae commonly known as Niepa bark tree. The drug had been extensively used in folklore practices and usage of the plant in main stream clinical practices is less. So giving a standardization and to justify its traditional usages preliminary phyochemical analysis had been done. The preliminary phytochemical analysis aims at analyzing the physico chemical property of drugs, their qualitative analysis, ash values, extractive values, moisture and volatile contents, estimation of Tannins and Phenols and HPTLC. Previous studies are available regarding the Qualitative Analysis of phytochemicals, tannin and total phenolic estimation. References regarding ash values, quantitative estimation of fiber, reducing sugar and total sugar, pH, cold and hot alcohol and water soluble extractives, moisture content were not available from previous research works On analyzing the phytochemical constituents present in the crude drug, the drug revealed the presence of alkaloids, flavonoids, saponins, carbohydrates, proteins, phenols, steroids, and tannins. This phytoconstituents present in the drug may be responsible for specific action of the drug. This preliminary phytochemical evaluation may be helpful to identify the potential of Guchakaranja and should be helpful in developing new formulations with additional therapeutic effect.

Quality evaluation of Terminalia billerica based on HPLC fingerprint and multi-components simultaneous determination / 中草药

Objective: To establish the HPLC fingerprint and to determin gallic acid, methyl gallate, corilagin, and ellagic acid in Terminalia billerica, in order to provide the scientific foundation for quality control of T. billerica. Methods: The analysis was performed on Atlantic T3 (250 mm × 4.6 mm, 5 μm) C18 column, mobile phase was acetonitrile-0.2% glacial acetic acid aqueous solution with gradient elution, flow rate was 1.0 mL/min, injection size was 20 μL column, and temperature was maintained at 30 oC. The common mode of T. billerica HPLC fingerprint was established, the hidden information was analyzed in the fingerprint by Chemometrics, and the components in T. billerica by HPLC-MSn and quantitative analysis characteristic peaks were identified. Results: There were 21 common peaks in the diagram and the similarity of the fingerprints was over 0.9 in all 11 batches. The information of the 18 common peaks in T. billerica was summarized by HPLC-MSn technology. The samples were broadly divided into three kinds by principal component analysis and clustering analysis. The five key compounds were verified by partial least squares discriminant analysis method in quantitative analysis, and identified that the No.12 peak was chabulagic acid, and the average recoveries were in the range of 97.24%-98.58%. Conclusion: The HPLC fingerprint method and content determination method are reliable, accurate, rapid, simple, and reproducible, and this study could control the quality of T. billerica.

Chemical constituents from Curculigo orchioids / 中成药

AIM To study the chemical constituents from Curculigo orchioides Gaertn..METHODS The n-butanol fraction of 95％ ethanol extract from C.orchioides was isolated and purified by silica,macroporous resin and Sephadex LH-20,then the structures of obtained compounds were identified by spectral data.RESULTS Seven compounds were isolated and identified as anacardoside (1),ocinol glucoside (2),3-(4-hydroxyl-3-methoxyphenyl)-propan-1,2-diol (3),benzyl-O-β3-D-glucopyranoside (4),icariside F2 (5),glucosyringic acid (6),calleryanin (7).CONCLUSION Compounds 3-7 are isolated from genus Curculigo for the first time.

Quality Evaluation of Terminalia billerica (Gaertn.) Roxb.Tannins Fraction from Different Habitats by HPLC Fingerprint Based on HPLC-MSn and Chemometrics / 世界科学技术-中医药现代化

This paper was aimed to establish the HPLC fingerprint of T.Billerica (Gaertn.) Roxb.tannin fraction.The analysis was performed on Atlantic T3 (250 mm × 4.6 mm,5 μm) C18 column.The mobile phase was acetonitrile-0.2％ glacial acetic acid aqueous solution with gradient elution.The flow rate was 1.0 mL·min-1.The injection size was 20 μL.The temperature was maintained at 30℃.Eleven batches of T.Billerica (Gaertn.) Roxb.tannin fraction of chromatographic data was analyzed by similarity of chromatographic data,SPSS software and SIMCA software.There were 20 common peaks in the diagram.The similarity analysis of 11 samples revealed that the similarities were between 0.832 and 0.973.Only the similarity of tannin parts from Xinjiang was below 0.9.The cluster analysis classified the tannin parts into 3 types,which shared the similar results as the principal components analysis (PCA).PLS-DA found that peak 1,13 and 14 may be the main chromatographic peaks to identify tannins fraction.The HPLC-MSn information of 14 compounds in T.billerica (Gaertn.) Roxb.was summarized.It was concluded that chemometrics analysis method can be used to analyze the HPLC fingerprint of T.billerica (Gaertn.) Roxb.tannin fraction.This method was rapid,simple,and reproducible.It can be used as one of the effective methods for the quality control of T.billerica (Gaertn.) Roxb.tannin fraction.

Chemical constituents from Curculigo orchioids / 中成药

AIM To study the chemical constituents from Curculigo orchioides Gaertn..METHODS The n-butanol fraction of 95％ ethanol extract from C.orchioides was isolated and purified by silica,macroporous resin and Sephadex LH-20,then the structures of obtained compounds were identified by spectral data.RESULTS Seven compounds were isolated and identified as anacardoside (1),ocinol glucoside (2),3-(4-hydroxyl-3-methoxyphenyl)-propan-1,2-diol (3),benzyl-O-β3-D-glucopyranoside (4),icariside F2 (5),glucosyringic acid (6),calleryanin (7).CONCLUSION Compounds 3-7 are isolated from genus Curculigo for the first time.

Study on flavonoids from Nicandra physaloides and their immune activities / 中草药

Objective: To investigate the flavonoids from the fruits of Nicandra physaloides and clarify the immune activities of the isolated compounds. Methods: The separations and purifications were taken by silica gel and ODS chromatogram columns as well as preparative HPLC, and the structural identification based on physicochemical property, 1H-NMR, and 13C-NMR as well as HR-MS data. And the immune activities were evaluated by mice splenic lymphocyte proliferation model induced by LPS. Results: Thirteen compounds were obtained from the butanol fraction of the 70% ethanol extract of N. physaloides fruits, which were identified as 3''-hydroxy-puerarin (1), puerarin-6″-O-glucoside (2), daidzein 8-C-[α-D-apiofranosyl-(1→6)]-β-D-glucopyranoside (3), puerarin (4), 3''-methoxyl-puerarin (5), chrysin 6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (6), oroxylin A-7-O-β-D-glucuronide methyl ester (7), daidzin (8), baicalin (9), quercetin-3-O-β-D-galactopyranoside (10), quercetin 3-rutinoside (11), kaempferol 3-rutinoside (12), and quercetin 3-glucoside (13), respectively. The results indicated that compounds 1-3, 7, 9-10, and 12-13 could inhibit the proliferation of mice spleen lymphocytes and 6 could promote the proliferation in the influence of mice spleen lymphocytes proliferation induced by LPS. Conclusion: Compounds 1-7 are isolated from the plants of Solanaceae for the first time, compounds 8-13 are obtained from the plants of Nicandra Adans. firstly. And the compounds 1-3, 6, 7, 9-10, and 12-13 possibly possess the potential immune activities.

Establishment of Silybum marianum cell suspension culture system and effects of different factors on silybin content / 中草药

To establish the suspension culture system of Silybum marianum cell and study the effects of different factors on silybin content. Orthogonal test was adopted to determine S. marianum cell suspension culture system and HPLC to silybin content.. The best growth cycle of cell suspension culture was defined when cultured in liquid MS medium with 6-BA 1.5 mg/L + NAA 1.0 mg/L + ZT 1.5 mg/L, light 12 h/d, pH value = 7, revolution = 110 r/min. The effects of salicylic acid, chitosan, and sodium nitroprusside on cell suspension culture were investigated. When the concentration of three impact factors were 0.05, 3, and 1 μ g/L, respectively, the growth amount of S. marianum cell reached the maximum, i. e. At the same time, the silybin content also reached the maximum. The established cell suspension culture system is suitable to use for the rapid propagation of S. marianum. Chitosan and sodium nitroprusside at proper concentration are benifit to the growth of suspensious cells and accumulation of silybin.

Study of quality testing method for seeds of Silybum marianum / 中草药

Objective: To study the quality testing method for the seeds of Silybum marianum, so as to provide the basis for the development of testing procedures and quality grading standard for the seeds of S. marianum. Methods: Refering to "The International Seed Testing Rules" and "China Crop Seed Testing Rules" to carry out the quality testing for the seeds of S. marianum. Results: The seed cleanliness was analyzed by winnowing method; The authenticity was identified by morphological appearance compared with 1000 grain weight by 1000 grain weight determination method; The germination conditions for seed germination were washed with running tap water before 2 h, the double filter paper was used as germination bed, and the seeds were incubated at 20 ℃, 8000 lx light, counting time for 4-14 d; The viability was determined by electrical conductivity method. Conclusion: The method is simple and reliable to the quality testing for the seeds of S. marianum.

Effect of sucrose and potassium nitrate on biomass and saponin content of Talinum paniculatum Gaertn. hairy root in balloon-type bubble bioreactor

Objective: To increase biomass and saponin production in hairy root culture of Talinum paniculatum Gaertn. (T. paniculatum) in balloon-type bubble bioreactor (BTBB). Methods: Hairy roots which were collected from leaf explants of T. paniculatum were infected by Agrobacterium rhizogenes strain LB510. The hairy roots were cultivated at 400 mL Murashige and Skoog liquid medium without growth regulator (MS0) in 1 000 mL BTBB. Each BTBB had 2 g hairy roots as initial inoculum and these cultures were treated with various concentrations of sucrose (3%, 4%, 5%, 6%w/v) and potassium nitrate (0.5, 1.0, 1.5 and 2.0 strength of MS medium). Cultures were maintained for 14 days. Fresh and dry weights of hairy roots at the end of culture were investigated. Results: Various concentrations of sucrose influenced the biomass accumulation of hairy roots. Maximum biomass was reached by MS medium supplemented with 6% sucrose and it was approximately threefold higher than control. Culture supplemented with po-tassium nitrate at 2.0 strength of MS0 could increase biomass accumulation of hairy roots until 0.14 g dry weight and it was almost threefold higher than control. However, the maximum saponin content was obtained by MS medium supplemented with 5%sucrose and 2.0 strength potassium nitrate of MS. Conclusions: Based on this research, those conditions can be used to produce biomass and saponin of hairy root of T. paniculatum in the large scale.

Comparison of Antioxidant Activity Between Phenolic and Nonphenolic Alkaloids in Nelumbo Nucifera Gaertn in vitro / 医药导报

Objective To compare the antioxidant activity between phenolic and nonphenolic alkaloids in Nelumbo Nucifera Gaertn.in vitro. Methods DPPH, ABTS, hydroxyl radical scavenging, super oxygen anion from oxidation, reducing power and beta carotene bleaching test methods were used to evaluate the antioxidant activity of the alkaloids. Results Half maximalinhibitory concentration ( IC50 ) of DPPH among total alkaloid, phenolic alkaloids and nonphenolic alkaloids was 21.89, 27.10 and 32.87 μg·mL-1 , respectively;IC50 of ABTS free radicals was 14.25, 20.55, 25.94μg·mL-1;The hydroxyl radical scavenging IC50 was 0.03, 0.03, 0.08 μg·mL-1; The auto-oxidation rate of super oxygen was 8.72×10-4, 5.87×10-4, 6.68× 10-4;The total alkaloid had the best reducing power, while the non-phennolic alakloids had the worst; Lipid inhibition rate of three alkaloids were 89.63%, 85.85% and 83.78% respectively. Conclusion Phenolic alkaloids are better than non-phenolic alkaloids in hydroxyl radical scavenging, reducing power and anti-lipid peroxidation, resulting in a promising prospect.

Effects of benzylbenzoate glucosides from Curculigo orchioides Gaertn. on retinoic acid induced osteoporosis in rats / 中国药学杂志

OBJECTIVE: To investigate the therapeutic effects of benzylbenzoate glucosides from Curculigo orchioides on retinoic acid induced osteoporosis in rats. METHODS: 3-Month-old female Sprague-Dawley rats were intragastrically given the retinoic acid 70 mg · kg-1 · d-1 for 2 weeks, and then treated with estrogen (E2, 1 mg · kg-1), total flavonoids from Epimedium folium (TF, 100 mg · kg-1 ), benzylbenzoate glucosides from Curculigo orchioides (COP, 6, 18 and 54 mg · kg-1) and Curculigo orchioides extracts (COE, 3 000 mg · kg-1) for 4 weeks, respectively. The bone mineral density of right femur were assayed by Dual Energy X-ray Absorptiometry. The levels of Ca, P, creatinine, ALP and TRAP in serum and urine were assayed with automatic biochemical analyzer. The levels of BGP and DPD in serum were measured with Elisa kit according to instruction. RESULTS: Treatment SD rats with retinoic acid for 2 weeks significantly decreased bone mineral density, increased the ratio of Ca and Creatinine in urine, and serum TRAP activity. Treatment with E2, TF and COP not only increased the bone mineral density, decreased the ratio of Ca and creatinine in urine and serum TRAP activity, but also regulated BGP level and increased ALP activity of serum in retinoic acid induced osteoporotic rats. CONCLUSION: Benzylbenzoate glucosides from Curculigo orchioides can decrease bone loss by inhibiting bone resorption and improving bone formation.

Effect of sucrose and potassium nitrate on biomass and saponin content of Talinum paniculatum Gaertn. hairy root in balloon-type bubble bioreactor

Objective: To increase biomass and saponin production in hairy root culture of Talinum paniculatum Gaertn. (T. paniculatum) in balloon-type bubble bioreactor (BTBB). Methods: Hairy roots which were collected from leaf explants of T. paniculatum were infected by Agrobacterium rhizogenes strain LB510. The hairy roots were cultivated at 400 mL Murashige and Skoog liquid medium without growth regulator (MS0) in 1. 000 mL BTBB. Each BTBB had 2 g hairy roots as initial inoculum and these cultures were treated with various concentrations of sucrose (3%, 4%, 5%, 6% w/v) and potassium nitrate (0.5, 1.0, 1.5 and 2.0 strength of MS medium). Cultures were maintained for 14 days. Fresh and dry weights of hairy roots at the end of culture were investigated. Results: Various concentrations of sucrose influenced the biomass accumulation of hairy roots. Maximum biomass was reached by MS medium supplemented with 6% sucrose and it was approximately threefold higher than control. Culture supplemented with potassium nitrate at 2.0 strength of MS0 could increase biomass accumulation of hairy roots until 0.14 g dry weight and it was almost threefold higher than control. However, the maximum saponin content was obtained by MS medium supplemented with 5% sucrose and 2.0 strength potassium nitrate of MS. Conclusions: Based on this research, those conditions can be used to produce biomass and saponin of hairy root of T. paniculatum in the large scale.

Emblica officinalis Stimulates the Secretion and Action of Insulin and Inhibits Starch Digestion and Protein Glycation In vitro.

Aim: Medicinal, edible and aromatic plants and natural products have been used worldwide for the management of diabetes mellitus. The aim of this study was to investigate the efficacy and mode of action of Emblica officinalis Gaertn. (Phyllanthaceae) used traditionally for treatment of diabetes. Study Design: Using multiple In vitro models; this study was designed to investigate the antidiabetes efficacy and mode of action of E. officinalis. Place and Duration of Study: School of Biomedical Sciences, University of Ulster, 2001- 2004 Results: E. officinalis aqueous extracts (AEs) stimulated basal insulin output and potentiated glucose-stimulated insulin secretion concentration-dependently in the clonal pancreatic beta cell line, BRIN-BD11 (p<0.001). The insulin secretory activity of plant extract was abolished in the absence of extracellular Ca2+ and by inhibitors of cellular Ca2+ uptake, diazoxide (p<0.001, n=8). Furthermore, the extract increased insulin secretion in depolarised cells and further augmented insulin secretion triggered by IBMX and tolbutamide. E. officinalis AE (1 mg/mL) displayed insulin mimetic activity (230%, p<0.001). Furthermore, it enhanced insulin-stimulated glucose transport in 3T3 L1 adipocytes by 460% (p<0.001). E. officinalis augmented also synergistically (p<0.001) insulin action, when co-incubated with insulin sensitizers; metformin (2.4-fold), vanadate (4.9-fold), tungstate (4.8-fold) and molybdate (6-fold). At higher concentrations (0.5-5 mg/mL), the extract also produced 8-74% (p<0.001) decrease in enzymatic starch digestion In vitro. E. officinalis AEs (1-50 mg/mL) inhibited protein glycation 44-87% (p<0.001). Conclusion: This study has revealed that water soluble bioactive principles in E. officinalis extract stimulate insulin secretion, enhance insulin action and inhibit both protein glycation and starch digestion. The former actions are dependent on the bioeffective component (s) in the plant being absorbed intact. Future work assessing the use of Emblica officinalis as adjunctive therapeutic nutraceutical or as a source of bioactive antidiabetic principles may provide new opportunities for the integrated management/prevention/reversal of diabetes.

Cloning of flavonoid 3'-hydroxylase gene from Fagopyrum tataricum and its tissue-specific expression under cold stress / 中草药

Objective: To clone and analyze the full-length of flavonoid 3'-hydroxylase (F3'H) gene from tartary buckwheat (Fagopyrum tataricum) and to express it in Escherichia coli. FtF3'H gene expression and anthocyanins accumulation is also to be analyzed in tartary buckwheat sprout under cold stress. Methods: Homology cloning and RACE method were used to obtain FtF3'H gene from flower buds of tartary buckwheat. The recombinant vector pET-30b (+)-FtF3'H was constructed and expressed in E. coli BL21 (DE3). Semi-quantitative RT-PCR was used to analyze FtF3'H gene expression when spectrophotometric method was used to determine anthocyanin content. Results: FtF3'H gene contains an open reading frame (1470 bp) encoding 489 amino acids and belongs to cytochrome P450 family. SDS-PAGE analysis of IPTG induced recombinant E. coli BL21 (DE3) showed that a predicted 54000 Da fusion protein was expressed in the culture. Cold stress significantly enhanced the expression level of FtF3'H and anthocyanin accumulation (P < 0.05). Conclusion: FtF3'H gene could be cloned from F. tataricum and efficiently expressed in E. coli. Under cold stress, FtF3'H gene may enhance its expression level to promote anthocyanin accumulation, by taking part in the process of cold-stress resistance of F. tataricum sprouts.

Effects of exogenous nitric oxide on physiology of seed germination and seedling growth of Silybum marianum under NaCl stress / 中草药

Objective: To investigate the effects of exogenous nitric oxide (NO) on the physiology of the seed germination and seedling growth of Silybum marianum under NaCl stress. Methods: The seeds of S. marianum were treated by sodium nitro prusside (SNP) at the concentration of 0.05-0.60 mmol/L under 0.7% NaCl stress. Some physiological indexes were measured, such as germination energy, germination rate, germination index, and vigor index of the seeds, and contents of malondialdehyde (MDA), photosynthetic pigment, osmosis substances, and the activities of the protective enzymes in leaves. Results: The seed germination and seedling growth of S. marianum were obviously inhibited under NaCl stress. Soaking seeds with 0.05-0.60 mmol/L SNP could alleviate the damage of NaCl stress. Under this treatment, the contents of photosynthetic pigment (including chlorophyll a, chlorophyll b, total chlorophyll, and carotinoid) and osmosis substances (including soluble sugar, soluble protein, and proline), and the activities of protective enzymes (including SOD, POD, and CAT) in the leaves were significantly increased, while the MDA content in the leaves was decreased. Conclusion: Soaking seeds with 0.05-0.60 mmol/L SNP could promote the salt resistance of the seeds and seedlings of S. marianum. The different cultivars of S. marianum differ in the sensitivity to SNP. The optimal concentration of SNP for the seed soaking of S. marianum with white and black skins is 0.10 and 0.40 mmol/L, respectively.