Molecular mechanism of Salvia miltiorrhiza Bunge and Carthamus tinctorius L. in the treatment of myocardial ischemia based on network pharmacology: a comparative study

Based on network pharmacology and molecular docking technology,the similarities and differences of anti-myocardial ischemia mechanism between Salvia miltiorrhiza Bunge(SM)and Carthamus tinctorius L.(CT)were studied.Firstly,based on the traditional Chinese medicine systems pharmacology(TCMSP),the related compounds of SM and CT were obtained,and the potential targets of these compounds were collected by the target fishing method.Genecards database was used to obtain targets related to myocardial ischemia.The cross targets of CT,SM,and myocardial ischemia were then selected,and the protein-protein interaction(PPI)network was constructed based on the STRING database.The cross targets were imported into the Metascape database for Gene Oncology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.Cytoscape software was used to build the topological network diagram of the drug-compound-target path.Finally,the binding ability of the active ingredient and the key target was verified by molecular docking.65 active ingredients and 38 potential targets were screened from SM,and 22 active ingredients and 58 potential targets were excavated from CT.Important targets common to SM and CT were TNF,IL6,VEGFA,AKT1,etc.The common enrichment pathways involved are fluid shear stress and atherosclerosis,IL-17 signaling pathway,pathways in cancer,and toxoplasmosis.The findings suggested that the two traditional Chinese medicines exerted the effect of myocardial ischemia through the characteristics of multiple targets,multiple pathways,and multiple compounds.

红花(Carthamus tinctorius L.)种子醇溶蛋白遗传多样性分析

利用A—PAGE（Acid—polyacrylamide gel electrophoresis）技术对来源于32个国家的53份红花材料进行了种子醇溶蛋白检测。结果表明，这些红花材料具有丰富的醇溶蛋白等位变异，共分离出15条迁移率不同的谱带。其中，每份材料可电泳出5-12条谱带，平均8．5条，所有材料有1条共有带。红花种子醇溶蛋白可作为评价红花遗传多样性的工具之一。不同材料的遗传相似系数（GS）变异范围为0．375—1．000，平均值为0．752。聚类分析结果表明，在GS值为0．752的水平上供试材料聚为6大类，其亲缘关系远近与地理来源关系不大。其中，来自中国的7份材料分别被聚在了3个大类中，表明中国红花种子醇溶蛋白遗传多样性比较丰富。

SRAP Analysis on Genetic Relationship of Carthamus tinctorius L Varieties in Xinjiang

[Objective] This study aimed to analyze the genetic relationship of the Carthamus tinctorius L（safflower） varieties in Xinjiang, and thus to provide a scientific basis for its protection and development. [Method] The genomic DNA of five safflower varieties from Xinjiang and one from Yunnan were analyzed and compared by SRAP molecular marker. [Result] Twelve pairs of SRAP primers were selected to amplify the genomic DNA of the six materials. Among the 171 clear DNA bands finally obtained, 93 were polymorphic, accounting for 54.4% of total. Genetic similarity coefficient of the six safflower varieties ranged from 0.60 to 0.92. [Conclusion]SRAP molecular marker is suitable for safflower varieties research and to guide the molecular breeding.

Protective effect of Salvia miltiorrhiza and Carthamus tinctorius extract against lipopolysaccharide-induced liver injury

AIM:To investigate the hepatoprotective effects and mechanisms of an extract of Salvia miltiorrhiza and Carthamus tinctorius in vivo.METHODS:C57BL/6J mice were randomly assigned to five groups and intraperitoneally administered 0.9% saline,Salvia miltiorrhiza and Carthamus tinctorius extract [Danhong injection(DHI),0.75 and 3 g/kg mixed extract] or reduced glutathione for injection(RGI,300 mg/kg) for 30 min before exposure to lipopolysaccharide(LPS,16 mg/kg). After intraperitoneal LPS stimulation for 90 min or 6 h,the mice were sacrificed by ether anaesthesia,and serum and liver samples were collected. Histological analysis(H&E) and terminal deoxynucleotidyl transferase-mediated d UTP nick end-labelling(TUNEL) staining were performed. Alanine transferase(ALT),aspartate transaminase(AST),total bilirubin(TBil),glutathione-S-transferase(GST),malondialdehyde(MDA),tumour necrosis factor(TNF)-α,interleukin(IL)-6,and caspase-3 levels were measured. Bax,Bcl-2,P-IκBα,IκBα,P-NF-κB p65,and NF-κB p65 protein levels were determined by Western blot. TNF-α,IL-6,caspase-3,Bax and Bcl-2 m RNA expression was measured by real-time reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:Hematoxylin-eosin staining and TUNEL results suggested that DHI(3 g/kg) treatment alleviated inflammatory and apoptotic(P < 0.01) injury in the liver of mice. DHI treatment dose-dependently blunted the abnormal changes in biochemical parameters such as ALT(72.53 ± 2.83 for 3 g/kg,P < 0.01),AST(76.97 ± 5.00 for 3 g/kg,P < 0.01),TBil(1.17 ± 0.10 for 3 g/kg,P < 0.01),MDA(0.81 ± 0.36 for 3 g/kg,P < 0.01),and GST(358.86 ± 12.09 for 3 g/kg,P < 0.01). Moreover,DHI(3 g/kg) remarkably decreased LPS-induced protein expression of TNF-α(340.55 ± 10.18 for 3 g/kg,P < 0.01),IL-6(261.34 ± 10.18 for 3 g/kg,P < 0.01),and enzyme activity of caspase-3(0.93 ± 0.029 for 3 g/kg,P < 0.01). The LPS-induced m RNA expression of TNF-α,IL-6 and caspase-3 was also decreased by DHI. Western blot analysis revealed that DHI antagonised LPS-stimulated decrease of Bcl-2 and increase of Bax protein expression. Furthermore,DHI inhibited LPS-induced IκBα and NF-κB p65 phosphorylation.CONCLUSION:DHI may be a multi-function protectant against acute hepatic injury in mice through its antiinflammatory,anti-oxidative and anti-apoptotic activities.

Progress of safflower (Carthamus tinctorius L.) regeneration through tissue culture

Background: Safflower regeneration through tissue culture has long been limited to low frequency and lack of an efficient protocol that suitable for most safflower cultivars. In past decades, researches had been carried out to investigate safflower regeneration through tissue culture and great progress had been made. Objective: To investigate factors that affect safflower regeneration through tissue culture principally. Methods: This article summarized available literatures about advancements in safflower regeneration, especially discussed factors affecting safflower tissue culture in detail. Results: Safflower regeneration was fairly hard than other congeneric plants, such as chrysanthemum. The genotype, seedling age, type of explants, medium components, plant growth regulators and other additives all had specific influences on safflower tissue culture. More deepgoing researches need to be undertaken to establish an effective safflower regeneration system.

Color spaces of safflower (Carthamus tinctorius L.) for quality assessment

Objective:In this study,safflower(Carthamus tinctorius L.)was taken as a representative example to examine the application of color characteristics to evaluate quality.Methods:A computer vision system was established for the objective and nondestructive assessment of color using image processing algorithms.Color parameters were investigated based on the RGB,L*a*b and HSV color spaces.The content of hydroxysafflor yellow A(HSYA),a major bioactive constituent of safflower,was determined by high-performance liquid chromatography.The relationship between HSYA content and color values was investigated by Pearson correlation analysis.A multiple linear regression model was established to predict the HSYA content from color values.Results:The red color and lightness of safflower were found to be significantly related to HSYA content.The prediction equation obtained by multiple regression was reliable with an R2 value of 0.805(P<.01).Conclusion:The results suggest that the computer vision technique could be used as a promising and non-destructive technology for color measurement and quality evaluation of CHM.

Safflower (Carthamus Tinctorius L.) a Potential Source of Drugs against Cryptococcal Infections, Malaria and Leishmaniasis

In this research we present that Carthamus Tinctorius L.(gen.Asteraceae,otherwise known as Safflower)(Fig.1)may contain agents active in Cryptococcal infections,malaria and Leishmaniasis,as treatment options are becoming scarce due to drug resistance development.Phytochemistry and pharmacological activities(antimicrobial,antimalarial,antileishmanial)of C.tinctorius L.were analyzed.The composition of volatile oil of safflower dried flowers was analyzed by gas chromatography-mass spectrophotometry with flame ionization detector(GC-FID)and in vitro sensitivity assays were performed to assess biological activity.8 known and 3 unknown compounds were detected in the extract(Fig.1).Then the Safflower ointment was manufactured and its acute toxicity study on rats was tested.The volatile oil of C.tinctorius L exhibited activity against Cryptococcus neoformans,Plasmodium falciparum and Leishmania donovani.Safflower volatile oil has anticryptococcal,antimalarial and antileishmanial effects.The prepared ointment had an excellent acute toxicity safety profile.

Assay for the developmental toxicity of safflower (Carthamus tinctorius L.) to zebrafish embryos/larvae

Objective:To explore the developmental toxicity and the potential toxicological mechanism of safflower (Carthamus tinctorius L.) on zebrafish embryos/larvae.Methods:Mortality,malformations and increased apoptosis induced by safflower were assessed in zebraflsh embryos from 6 to 96 hours post-fertilization.Enzymes and genes in the anti-oxidative and apoptotic pathways were also assayed.Results:The lethal concentration 50 of safflower to zebrafish embryos was 345.6 mg/L.Hatching inhibition,abnormal spontaneous movement,depressed heart rate,pericardial edema,yolk sac edema,abnormal head-trunk angle,inhibition of melanin release,enlarged yolk,and short body length were observed in safflower-treated zebrafish.Additional apoptotic cells mainly appeared around the heart.Safflower exposure changed the activities of defense enzymes (SOD T,CAT↑,MDA↑,GPX without trend),increased MDA content,decreased caspase-3 activity,and altered mRNA levels of related genes (ogg1 ↓,p53↓,Cu/Zn-sod↑,Mn-sod↓,cat↓,gpx↑).Conclusion:Safflower exhibits developmental toxicity for zebrafish embryos/larvae.The developing heart was speculated as the target organ of toxicity.Oxidative stress and increased apoptosis have roles in the developmental toxicity of safflower.This article provides a novel method to research the teratogenicity and possible mechanisms of toxicity of traditional Chinese medicines that are prohibited or contraindicated in pregnant women.

Influence of Meta-Topolin on Efficient Plant Regeneration via Micropropagation and Organogenesis of Safflower (<i>Carthamus tinctorius</i>L.) cv. NARI-H-15

The effect of meta-Topolin (mT) was assessed to develop a reliable protocol for efficient plant regeneration of safflower (Carthamus tinctorius L.) cv. NARI-H-15. For micropropagation, 7 - 9 days old shoot-tip explants cultured on MS basal medium supplemented with 3.0 mg/L meta-Topolin (mT) + 0.5 mg/L CPPU showed 97.7% adventitious shoot formation (42.4 shootlets) than node after 45 days of culture. For organogenesis, the seedling explants of immature leaf cultured on 1.5 mg/L CPPU or 1.5 mg/L NAA fortified medium produced high amount of callus than cotyledon and stem calli after 60 days of culture. However, MS basal medium fortified with 4.0 mg/L mT + 1.5 mg/L CPPU was found beneficial to stimulate 100% organogenic response (74.7 shootlets) from immature leaf calli than cotyledon and stem derived calli after 45 days of culture. The healthy plantlets obtained from micropropagation and organogenesis process cultured on 1/4 MS basal salts, 1.5% sucrose (w/v) and 0.8% agar (w/v) medium supplemented with NAA (1.5 mg/L) and mT (0.1 mg/L) produced maximum of 96% (12.8 rootlets) and 84% (7.3 rootlets) adventitious rooting, respectively than mT and CPPU tested medium. However, maximum of 67% and 42% survival rate was noticed when in vitro raised plants from micropropagation and organogenesis were hardened in pots containing soil mix and maintained under green house condition. This optimized regeneration protocol might be helpful in regeneration of new genotypes and cultivars of safflower to improve agronomic traits through in vitro selection process and Agrobacterium-mediated genetic transformation system.

Phosphorus Use Efficiency of Safflower （Carthamus tinctorius L.） and Sunflower （Helianthus annuus L.） Studied in Nutrient Solution

Safflower represents an important oil crop internationally and may have a production potential under low input conditions, but its putatively high phosphorous use efficiency is not sustained. This study aims to directly compare safflower with sunflower in terms of phosphorus use efficiency in nutrient solution under controlled conditions. Growth of both species responded strongly to increasing P supply. Safflower recovers less proportion of added P than sunflower. External P requirement （（g P supply （100 g dry matter （DM） produced）~） was higher in safflower than sunflower. The efficiency of the crops for DM production based on accumulated P （mg P potl, efficiency ratio）, and P concentration in DM （（mg P （g DM）＇I）, utilization index） were interpreted using Michaelis-Menten kinetics as growth response curves. Accordingly, Km constant was lower in sunflower compared to safflower in terms of utilization index, but both were similar in terms of efficiency ratio. High Km constant in safflower in terms of utilization index indicates the high P concentration in tissues to produce 50% of potential maximum DM, consequently less efficient crop. Utilization efficiency contributed more than uptake efficiency in overall PUE in the efficient cultivar and could be the cause of its superiority in PUE. It can be concluded that safflower has a high requirement for P with respect to growth, sunflower is more efficient in terms of uptake and utilization of P at optimal and sub-optimal P supplies indicating that safflower can not be considered a low nutrient input crop compared to sunflower with respect to phosphorus.

油用型红花(Carthamus tinctoris L.)种子醇溶蛋白遗传多样性分析

利用A-PAGE（Acid-polyacrylamide gel electrophoresis）技术对来源于不同国家的79份油用型红花材料醇溶蛋白位点进行检测。结果表明，油用型红花醇溶蛋白位点存在丰富的变异类型，共分离出15条迁移率不同的谱带，每份材料具有3—12条不等，平均8．5条。材料间平均遗传相似系数（GS）为0．5303，变幅为0．1333—1．000，且各洲间遗传多样性大于洲内遗传多样性。在GS值为0．512的水平上，供试材料聚为六大类，聚类结果表明，红花醇溶蛋白图谱类型与其地理分布有一定的相关，但不显著。

Phytochemistry,Pharmacology and Medicinal Properties of Carthamus tinctorius L.

Carthamus tinctorius L. is commonly known as Safflower. C. tinctorius extracts and oil are important in drug development with numerous pharmacological activities in the world. This plant is cultivated mainly for its seed, which is used as edible oil. For a long time C. tinctorius has been used in traditional medicines as a purgative, analgesic, antipyretic and an antidote to poisoning. It is a useful plant in painful menstrual problems, post-partum hemorrhage and osteoporosis. C. tinctorius has recently been shown to have antioxidant, analgesic, anti-inflammatory and antidiabetic activities. Carthamin, safflower yellow are the main constituents in the flower of C. tinctorius. Carthamidin, isocarthamidin, hydroxysafflor yellow A, safflor yellow A, safflamin C and luteolin are the main constituents which are reported from this plant. Caryophyllene, p-allyltoluene, 1-acetoxytetralin and heneicosane were identified as the major components for C. tinctorius flowers essential oil. Due to the easy collection of the plant and being widespread and also remarkable biological activities, this plant has become both food and medicine in many parts of the world. This review presents comprehensive analyzed information on the botanical, chemical and pharmacological aspects of C. tinctorius.

Cloning,expression and activity analysises of chalcone synthase genes in Carthamus tinctorius

Objective:Flavonoids are the bioactive compounds in safflower(Carthamus tinctorius),in which chalcone synthase(CHS)is the first limiting enzyme.However,it is unclear that which chalcone synthase genes(CHSs)are participated in flavonoids biosynthesis in C.tinctorius.In this study,the CHSs in the molecular characterization and enzyme activities were investigated.Methods:Putative chalcone biosynthase genes were screened by the full-length transcriptome sequences data in C.tinctorius.Chalcone biosynthase genes in C.tinctorius(CtCHSs)were cloned from cDNA of flowers of C.tinctorius.The cloned gene sequences were analyzed by bioinformatics,and their expression patterns were analyzed by real-time PCR(RT-PCR).The protein of CtCHS in the development of flowers was detected by polyclonal antibody Western blot.A recombinant vector of CtCHS was constructed.The CtCHS recombinant protein was induced and purified to detect the enzyme reaction(catalyzing the reaction of p-coumaryl-CoA and malonyl-CoA to produce naringin chalcone).The reaction product was detected by HPLC and LC-MS.Results:Two full-length CtCHS genes were successfully cloned from the flowers of safflower(CtCHS1 and CtCHS3),with gene lengths of 1525 bp and 1358 bp,respectively.RT-PCR analysis showed that both genes were highly expressed in the flowers,but the expression of CtCHS1 was higher than that of CtCHS3 at each developmental stage of the flowers.WB analysis showed that only CtCHS1 protein could be detected at each developmental stage of the flowers.HPLC and LC-MS analyses showed that CtCHS1 could catalyze the conversion of p-coumaryl-CoA and malonyl-CoA substrates to naringin chalcone.Conclusion:CtCHS1 is involved in the biosynthesis of naringin chalcone in safflower.

化肥减量配施微生物菌肥对膜下滴灌红花生长发育及产量的影响

【目的】研究化肥减量配施微生物菌肥对膜下滴灌红花生长发育及产量的影响。【方法】2020~2021年在新疆塔城地区裕民县设置2年定位施肥试验,采用裂区试验设计,设6个处理:(1)对照CK:不施肥;(2)CF:复合肥20 kg/667m^(2);(3)OF:微生物菌肥20 kg/667m^(2);(4)CF+25M:化肥15 kg/667m^(2)(化肥减量25%)+微生物菌肥5 kg/667m^(2);(5)CF+37.5M:化肥12.5 kg/667m^(2)(化肥减量37.5%)+微生物菌肥7.5 kg/667m^(2);(6)CF+50M:化肥10 kg/667m^(2)(化肥减量50%)+微生物菌肥10 kg/667m^(2)。研究不同施肥处理对红花农艺性状、干物质积累与分配和产量形成的影响。【结果】处理CF+25M和CF+37.5M有利于红花的生长发育,可以显著增加红花株高、分枝数、叶片数和叶绿素含量等,并能促进红花干物质积累,调节干物质分配,协调改善红花产量构成因素,从而增加红花花丝和籽粒产量。其中以CF+37.5M处理下红花综合性状表现最优,其单株果球数、千粒重、花丝产量和籽粒产量分别比CF处理显著提高了87.85%、12.29%、11.42%和15.78%,同时株高、根长和叶绿素含量亦均达到最高水平。【结论】化肥减量配施微生物菌肥CF+37.5M:化肥12.5 kg/667m^(2)(化肥减量37.5%)+微生物菌肥7.5 kg/667m^(2)最优,能有效降低化肥施用量,促进红花生长发育,从而提高肥料利用率。

水晶丹外敷治疗难治性痛风急性发作的临床观察

目的探索水晶丹外敷治疗难治性痛风急性发作的疗效与机制。方法选取2021年6月至2022年12月四川省医学科学院·四川省人民医院(电子科技大学附属医院)中西医骨伤科门诊收治的58例难治性痛风急性发作患者,随机分为对照组和试验组,对照组给予依托考昔+健康教育,试验组在对照组治疗基础上加水晶丹外敷治疗。分别于干预前和干预后第1、3、5天通过测皮温、膝关节周径、视觉模拟评分量表(VAS)评分、美国特种外科医院(HSS)量表观察其临床疗效;通过测滑膜液细胞因子IL-1β、TNF-α、TGF-β1浓度探究其抗炎机制。结果与对照组比较,试验组皮温、膝关节周径和VAS评分均明显较低(P<0.05),HSS量表评分较高(P<0.05),滑膜液IL-1β、TNF-α浓度均明显较低(P<0.05),TGF-β1浓度升高(P<0.05)。结论水晶丹外敷能通过抑制细胞致炎因子IL-1β、TNF-α浓度以及提高抗炎因子TGF-β1浓度表达途径来治疗难治性痛风急性发作。

水氮耦合对膜下滴灌红花生长发育及产量的影响

【目的】研究水氮耦合对膜下滴灌红花生长发育、干物质积累量和产量的影响。【方法】采用裂区试验设计,以灌水量水平为主区。播前施基肥磷酸二铵(总养分64%,N-P_(2)O_(5)=18-46)150 kg/hm^(2),氧化钾(30%)75 kg/hm^(2)。设灌水量(W)和施氮量(N)2个因素,3个灌水量水平:W_(1):1200 m^(3)/hm^(2)(低灌量)、W_(2):1500 m^(3)/hm^(2)(中灌量)和W_(3):1800 m^(3)/hm^(2)(高灌量);3个施氮量水平:N_(0):不施氮(空白)、N_(1):300 kg/hm^(2)(传统施氮量)和N_(2):600 kg/hm^(2)(高施氮量)。测定不同处理下红花的农艺性状、干物质积累量和产量,评估不同水氮耦合处理对红花生长发育和产量影响。【结果】适宜的灌溉量和施肥量对红花生长发育有显著的促进作用,W_(2)N_(0)和W_(2)N_(1)处理下红花的株高、分枝数和叶片数等农艺性状均显著高于其他处理,其中W_(2)N_(1)处理下红花干物质积累速率及积累量、单株果球数和每果粒数均达到最高水平,花丝产量和籽粒产量在W_(2)处理下均比W_(1)和W_(3)处理显著增加了8.09%和20.84%,但在N_(1)和N_(2)处理下差异较小,却均比N_(0)处理显著增加了10.69%和25.66%。【结论】中灌量和传统施肥量处理W_(2)N_(1)(灌溉量1500 m^(3)/hm^(2)及施肥量300 kg/hm^(2))最优,可以促进红花的生长发育、提高水肥利用效率、增加干物质积累、协调产量构成因素,达到高产优质。

种植密度和行距对膜下滴灌红花生长发育及产量的影响

【目的】研究不同种植密度和行距对膜下滴灌红花生长发育和产量的影响,为构建红花高光效种植模式提供参考。【方法】2020~2021年在新疆塔城地区裕民县设置田间试验,采用两因素随机区组设计,种植密度设4个处理:1.0×10^(4)株/667m^(2)(A_(1))、1.5×10^(4)株/667m^(2)(A_(2))、2.0×10^(4)株/667m^(2)(A_(3))和2.5×10^(4)株/667m^(2)(A_(4)),行距设2个处理:20 cm(B_(1))和30 cm(B_(2)),分析不同种植密度和行距对膜下滴灌红花农艺性状、干物质积累和产量的影响。【结果】在一定种植密度范围内,红花株高、叶绿素含量、干物质积累量、产量等指标均随种植密度的增加而逐渐增加,单株果球数、每果粒数和千粒重则随着种植密度的增加而降低,而不同行距则对红花的各指标影响较小。A_(3) B_(1)和A_(3) B_(2)处理下红花的综合指标和产量均显著优于其他处理,A_(3)处理下花丝产量分别比A_(1)、A_(2)和A_(4)处理增加了15.73%、11.23%和7.97%,籽粒产量分别比A_(1)、A_(2)和A_(4)处理增加了18.75%、11.96%和14.50%。【结论】种植密度(2.0×10^(4)株/667m^(2)(A_(3)),20 cm(B_(1))或30 cm(B_(2)),等行距)为红花高产高效的栽培模式,能协调红花生长,显著提高红花花丝和籽粒产量。

新疆红花的产量等级划分及增产措施

采用文献整合分析的方式,利用联合国粮食及农业组织官网下载的世界及中国的红花子粒产量数据,调查了新疆红花生长指标、产量及增产措施效果。结果表明,新疆红花株高、茎粗、分枝高、果球直径的均值分别为88.97、1.58、36.45、2.50 cm,单株平均果球数为15.71个,平均千粒重为43.61 g,花丝产量为152.00~1 350.00 kg/hm^(2),平均为421.06 kg/hm^(2),子粒产量为540.00~2 913.00 kg/hm^(2),平均为1 699.35 kg/hm^(2)。相关分析表明,红花花丝产量与各生长指标间存在不同程度的相关性,子粒产量未呈显著相关性。2016—2021年全球与中国红花子粒产量均值分别为949.4 kg/hm^(2)和1 458.5 kg/hm^(2),将子粒产量1 800 kg/hm^(2)及以上作为高产田,1 350~1 800 kg/hm^(2)作为中高产田,900~1 350 kg/hm^(2)作为中产田,900 kg/hm^(2)以下作为低产田。合理密植、灌溉、施肥等措施均能提高红花产量。适当提高种植密度可提高产量31%~54%,合理灌溉可增加产量33%~92%,而合理施肥可提高产量约1倍。新疆红花产量变异较大,有必要进一步探索红花高产稳产栽培技术体系。

红花CtWRKY7和CtWRKY72基因的克隆及功能分析

根据前期对红花(Carthamus tinctorius Linn.)WRKY基因家族的鉴定结果,利用RT-PCR技术克隆了红花CtWRKY7和CtWRKY72基因,其蛋白质编码序列(CDS)长度分别为600和861 bp,全长序列均含有3个外显子和2个内含子。CtWRKY7和CtWRKY72蛋白分别含有199和286个氨基酸残基,且二者均无跨膜结构和信号肽,定位于细胞核,二级结构主要由无规卷曲组成,三级结构较为松散。多序列比对及系统进化分析结果显示:CtWRKY7与菊花〔Chrysanthemum×morifolium(Ramat.)Hemsl.〕CmWRKY15和莴笋(Lactuca sativa var.angustata Irish ex Bremer)LsWRKY18的亲缘关系最近,均属于Ⅱa类WRKY;CtWRKY72与拟南芥〔Arabidopsis thaliana(Linn.)Heynh.〕AtWRKY70和AtWRKY54的亲缘关系最近,均属于Ⅲ类WRKY。启动子分析结果显示:CtWRKY7和CtWRKY72的启动子均含有多种逆境胁迫应答元件,且CtWRKY7的启动子含有W-box,而CtWRKY72的启动子无W-box。基因表达分析结果显示:CtWRKY7和CtWRKY72在红花的不同组织中均有表达,其中,CtWRKY7在根中表达量最高,CtWRKY72在叶中表达量最高;且二者均在花组织形成过程中表达量逐渐下降,在花开放过程中表达量逐渐上升。此外,CtWRKY7和CtWRKY72均响应多种逆境胁迫和外源激素诱导,但表达模式不同,总体上上调表达,仅CtWRKY7在外源ABA诱导下下调表达。综上所述,CtWRKY7和CtWRKY72可能参与红花花早期发育及衰老过程,CtWRKY7可能存在自我调节或与其他WRKY蛋白存在交叉调节;并且,CtWRKY7和CtWRKY72能响应多种逆境胁迫,但作用机制不同。

红花中芳香苷类化学成分研究

目的:研究菊科红花属植物红花Carthamus tinctorius L.的化学成分。方法:采用硅胶、MCI gel CHP-20、Sephadex LH-20、Toyopearl HW-40C以及半制备HPLC对红花70%含水丙酮提取物进行分离纯化,根据理化性质及波谱数据鉴定化合物的结构。结果:从红花中分离得到15个芳香苷类化合物,分别鉴定为:苄基-β-D-葡萄糖苷(1)、苄基-O-芸香糖苷(2)、苯乙醇芸香糖苷(3)、3-(4-O-β-D-吡喃葡萄糖基苯基)丙酸甲酯(4)、carthamoside B5(5)、junipediol A-8-O-β-D-glucopyranoside(6)、7-O-β-苯甲酰芸香糖苷(7)、紫丁香苷(8)、4-烯丙基-2,6-二甲氧基苯基葡萄糖苷(9)、2,6-二甲氧基-4-[(1E)-丙-2-烯基]苯基-O-芸香糖苷(10)、4-O-β-D-吡喃葡萄糖基反式苯丙烯酸(11)、4-O-β-D-吡喃葡萄糖基顺式苯丙烯酸(12)、1-(4-O-β-D-吡喃葡萄糖苷)苯甲酸(13)、4-(2-氨基乙基)苯基-1-O-β-D-吡喃葡萄糖苷(14)、(+)-南烛木树脂酚-3α-O-β-D-吡喃葡萄糖苷(15)。结论:其中,化合物1~4、10、13~15为首次从红花属植物中分离得到。