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.

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.

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.

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.

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.

Simultaneous determination of four nucleosides in Carthamus tinctorius L.and Safflower injection using high-performance liquid chromatography

We quantitatively determined four nucleosides, including cytidine, uridine, guanosine, and adenosine, in Carthamus tinctorius L. and Safflower injection. Separation was performed on a Zorbax Eclipse XDB-18 column using a gradient elution with mobile phases of 0.05% trifluoroacetic acid （TFA） aqueous solution （A） and methanol （B）. The assay was carried out at a flow rate of 1 mL/min at 25 ℃ with detection at 260 nm. Cytidine, uridine, adenosine and guanosine showed good linearity in the ranges of4.02-503μg/mL （r2= 0.9998）, 9.38-1407 μg/mL （rz = 0.9999）, 80.6-8060μg/mL （r2 = 0.9999） and 2.10---630μg/mL （r2 = 0.9987） with average recoveries of 97.2%, 94.5%, 98.6% and 108.6%, respectively. The contents of cytidine, uridine, adenosine and guanosine in different Carthamus tinctorius L. and Safflower injection were significantly different. This is the first report on the quantitative determination of nucleosides in Carthamus tinctorius L. and Safflower injection.

红花(Carthamus tinctorius)热激转录因子CtHSFA9的克隆及表达分析

根据课题组前期转录组文库信息获得的红花热胁迫相关转录因子Unigene的全长序列,设计引物并克隆该基因,采用生物信息学方法对其氨基酸序列的理化性质、系统发育、蛋白质亚细胞定位、结构域及跨膜区域等进行预测和分析。此外,试验还采用RT-qPCR方法分析该基因在红花各组织包括根、茎、叶、花及4个不同发育时期的种子中的表达情况。结果发现,所获序列全长1326 bp,可编码441个氨基酸,相对分子质量50.41 kD。其中,最丰富的氨基酸为亮氨酸(Leu),带正电残基64个,带负电残基75个。信号肽及亚细胞定位揭示该基因不存在明显信号肽且该基因属细胞核定位。SMART分析表明该基因具有典型HSF结构域。进化树分析表明,该序列与热激转录因子HSFA9基因相似度较高,与刺菜蓟的亲缘关系最近,因此可将其命名为CtHSFA9。荧光定量PCR分析表明CtHSFA9基因在红花根组织中相对表达量最高,叶次之,花中最低。在不同发育时期种子中,以花后18 DAF(Day after flower)中表达量最高,22 DAF次之,在14 DAF表达量最低。本研究初步明确了红花CtHSFA9基因的结构特点和进化关系,其属于HSF热激转录因子家族,可为后续开展红花逆境相关转录因子的机理研究做铺垫,同时也为植物抗逆性状的改良提供研究基础。

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.

Flavonoids from Carthamus tinctorius

Two new flavonoids, (2S)-4′ ,5-dihydroxyl-6,7-di-O-β-D-glu-copyranosylflavanone (1) and 6-hydroxykaempferol 6,7-di-O-β-D-glucopyranoside (2), were isolated fromCarthamus tinctorius . Their structures were elucidated by spectroscopic means including 2D NMR,ESIMS and CD.

Isocartormin, a novel quinochalcone C-glycoside from Carthamus tinctorius

A new semi-quinonechalcone C-glycoside isocartormin along with cartormin and saf fl omin C were isolated from the water extract of Carthamus tinctorius L. The structure of isocartormin was determined by extensive analysis of HR-MS, 1D-and 2D NMR data, and by comparison with those of cartormin reported previously by our group. Isocartormin was identified as a diastereoisomer of cartormin with a reverse configuration at C-18.

UGT88B2: A promiscuous O-glycosyltransferase from Carthamus tinctorius

Objective:In order to obtain new glycosyltransferases with highly efficient catalysis,the glycosyltransferases from Carthamus tinctorius which contains diverse types of glycosides were mined.Methods:A new glycosyltransferase gene(UGT88B2)with full length was obtained by PCR and further transformed into Escherichia coli for heterologous expression.The catalytic activity of recombinant UGT88B2 was determined by HPLC-MSn.The structures of representative catalytic products were elucidated by MS and NMR.Results:UGT88B2 exhibited catalytic promiscuity and various patterns in glycosylation of flavonoids with high efficiency.Conclusion:A new glycosyltransferase named UGT88B2 was successfully mined and can be employed as enzymatic tools in glycosylation of flavonoids.

Regeneration of Carthamus tinctorius from Jimsar

Objective To investigate the factors significantly affecting the shoot buds regeneration of Carthamus tinctorius from Jimsar （CTJ）. Methods Through tissue culture experiments, the influences of culture temperature, illuminance, relative humidity, explants type, seedlings age, and plant growth regulators added in culture media on in vitro regeneration of CTJ were investigated. Results The culture temperature was set at 24 ℃ in daytime and 16 ℃ at night, illuminance at 9000 lx, and 60% relative humidity were suitable for CTJ regeneration. Cotyledons excised from 6-8 d old seedlings were more responsive than even-aged euphyll, hypocotyl, and root explants because of inducing adventitious buds. The highest percentage of regenerated shoots （79. 1%） with about five adventitious buds per responding explant was obtained from MS basal medium containing 12.0 mg/LTDZ, 2.5 mg/L IBA, and 1.5 mg/L 2-ip （No. 14 medium）. Regenerated shoot buds （80%） could elongate successfully after 1-2 weeks transferred to shoot elongation medium. Conclusion Suitable factors of shoot buds regeneration for CTJ were determined. It may also make a useful reference for regeneration researches of other C. tinctorius varieties in China.

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。

Potential for Safflower Cultivation under Temperate Conditions

Safflower is a Mediterranean-subtropical oil crop, with temperate climate. To assess the suitability of safflower may have certain production potential in organic farming in regions cultivation under temperate climate in low input organic farming, ten cultivars were grown in two years experiment in Lindhoft-Kiel in Northern Germany in clay/sand soil. Pearson correlations reveal that oil yield exhibited tight positive correlations with plant height, plant dry matter, capitula plant-1, achenes capitulum-1 and thousand achene mass （TAM）, but was not correlated with days to full flowering or oil concentration. Yield component analysis reveals that achenes capitulum1 had the major influence on the variation of oil yield of all studied cultivars, followed by capitula plant1 and TAM, while the oil concentration had slightly negative influence, and the plant density was negatively associated with oil yield. Although two years field experiment is not enough to analyse the performance of safflower adaptability to certain region, it can be concluded that the most adaptive cultivars are PI-209286 originated from Romania, and cultivar CART-19/89 originated from Poland, while the less adaptive cultivars were DO-13/03 and DO-15/03 with German origin in addition to the Canadian cultivar PI-572475/Saffire. The most striking yield component that contributes to the high yield is achene capitulumt, which can be used as a base for selecting cultivars adaptive to humid cold conditions. The growth period between full flowering and harvest influencing this yield component must be given attention to avoid low temperature and rainfall by tuning sowing date to improve yield at these conditions.

Effects of Plant Population on Growth, Development and Oil Yield of Safflower

Two field trials were clone to evaluate the effects of plant density on the growth, development and yield of safflower. The results showed that plant density and season of growth had significant （P ≤ 0.01） effects on growth, development, yield components, yield and oil content of safflower. Increasing safflower plant density from 100,000 to 250,000 plants ha^- significantly reduced plant height （13.2%-21.3%）, branch number plant^-1 （37%-54.7%）, leaf number plant^-1 （39%-39.2%）, leaf area （19.5%-53%）, plant spread （39.6%-54.4%）, root length （28.1%-54.4%）, plant biomass （17%-50%）, capitula size （12%-12.7%）, capitula number plant^-1 （39.5%-50.5%）, seed number capitula~ （39%-45%）, capitula weight （3.3%-3.6%）, seed yield （67.9%-69.8%） and seed oil content （14.7%-20.8%）. The reduction in vegetative growth, yield components, yield and oil content of safflower due to increased plant density was attributed to inter and intra-plant competition for light, nutrients and water necessary for growth and development. The differences between winter and summer grown safflower were attributed to difference in day and night temperature （DIF） and the average daily temperature which were optimum for safflower growth in winter. It was concluded that under Botswana conditions or in semi-arid areas, safflower should be planted at 50 cm × 20 cm or wider in order to maximize yield and oil content and allow the plants to express their maximum genetic potential.

The Main Insect Pests of Safflower on Various Plant Parts in Iran

Safflower （Carthamus tinctorius） has its own limited factors, especially insect pests. There are several injurious insect pests reported from various safflower growing areas in Iran, causing severe and economical losses on various parts of safflower plants. They are safflower Budfly and capsulfly （Acanthiophilus helianthi）, aphid （Uroleucon carthami）, green leafhopper （Empoasca decipiens）, ground bug （Oxycarneus pallens）, grampod borer and capsule borer or bordered straw （Heliothis （and or Heliocaverpa） peltigera）, red spider mite and two spotted spider mite （Tetranychus urtica）, beet armyworm （Spodoptera exigua）, scarab beetle （Tropinota （and or Epicometis） hirta）, Egyptian cotton leaf （Spodoptera littoralis）, cotton boll worm （Heliothi absouleta） and so on. The other reported safflower insect pests from other countries are wireworms （Limonius spp.）, lygus bugs （Lygus hesperus）, flower thrips （Eastern or Western） or onion thrips （Thrips tabaci）, and seed corn maggots （Delia platura）, sunflower moths （Helianthus annuus） and Caterpillar （Perigaea capensis） are the serious pests of this crop. The other reports showed that some stored-product insects are capable of establishing infestations in safflower seeds and seed meal under suitable conditions. Only Lasiaderma serricorne, Stegobium penliceum, and the four species of Trogodema were to be as the infectious pests. Safflower is also attractive to several beneficial insects, including lady beetles and lacewings that may provide insect control in surrounding fields.

红花SCoT-PCR反应体系的建立与优化

为构建红花最佳扩增体系,为以后研究红花遗传多样性和目的基因标记提供了科学依据。本研究采用L_(16)(4^(5))正交试验和单因素试验相结合的方法对红花SCoT-PCR反应体系中的引物,DNA、dNTPs、PCR Buffer (含Mg^(2+))、Taq DNA聚合酶进行优化,每个因素都设为四水平。结果表明,五个影响因素中dNTPs的影响最大,红花最佳反应体系为引物0.6 mmol/L,DNA 60 ng,dNTPs 0.25 mmol/L,Taq DNA聚合酶1.5 U,PCR Buffer (含Mg^(2+)) 2μL,ddH_(2)O补足至20μL。选用引物S15对八种红花资源对扩增体系进行验证,结果显示扩增条带清晰,多态性好,表明该体系适合红花分子标记研究。

HPLC fingerprint-oriented preparative separation of major flavonoids from safflower extract by preparative pressurized liquid chromatography

The aim of this study was to rapidly isolate the major effective flavanoids from the extract of safflower（Carthamus tinctorius） using ODS medium pressure liquid chromatography（MPLC） and semi-preparative HPLC, guided by a developed fingerprint. Twelve compounds were isolated and their structures were elucidated as kaempferol 3-O-β-D-rutinoside（1）, kaempferol 3-O-β-D-glucoside（2）, rutin（3）, quercetin 3-O-β-D-glucoside（4）, 6-hydroxykaempferol 3,6,7-tri-O-β-D-glucoside（5）, 6-hydroxykaempferol 3-O-β-D-glucoside（6）, 6-hydroxykaempferol 6,7-di-O-β-D-glucoside（7）, 6-hydroxykaempferol 3-O-β-Drutinoside（8）, 6-hydroxykaempferol 3,6-di-O-β-D-glucosyl 7-O-β-D-glucuronide（9）, isosafflomin C（10）, safflomin C（11） and hydroxysafflor yellow A（12） by spectroscopic analysis and comparing with the literature. Our results demonstrated that preparative pressurized liquid chromatography combined with HPLC fingerprint guide is an efficient tool to isolate the target compounds quickly.