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.

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.

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.

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。

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.

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.

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.

红花二氢黄酮醇4-还原酶基因CtDFR1的克隆与分析

为了探索二氢黄酮醇4-还原酶(dihydroflavonol 4-reductase,DFR)基因在红花(Carthamus tinctorius L.)花色形成中的作用机制,本研究在分析红花转录组数据的基础上,以红花二氢黄酮醇4-还原酶为研究对象,通过PCR方法从红花的管状花中克隆得到一个红花DFR基因家族的成员,命名为CtDFR1。测序结果表明,该基因开放阅读框包含1029 bp,编码342个氨基酸;为稳定的亲水性蛋白,具有二氢黄酮醇4-还原酶结构域,属PLN02650超基因家族。多重序列比对及进化树分析表明,红花与矢车菊、斑点矢车菊DFR的氨基酸相似性较高,分别为95.06%和95.03%。荧光定量PCR结果显示,CtDFR1基因在花中表达量最高,其次是叶和苞片,在根和茎中检测不到该基因的表达。对不同花色红花品种的表达分析表明,CtDFR1基因在橙色和白色红花中表达量最高,在黄色和红色中表达量较低,预示了CtDFR1与红花的花色差异具有相关性。本研究获得了CtDFR1基因的序列及表达情况,为深入研究CtDFR1基因的功能提供了科学依据。

红花ω-3脂肪酸脱氢酶基因FAD3的克隆及表达分析

亚麻酸属植物中的多不饱和脂肪酸,其由微粒体和质体类两种不同类型的ω-3脂肪酸脱氢酶催化合成的,一般认为微粒体ω-3型脂肪酸脱氢酶基因(FAD3)是控制种子油中α-亚麻酸合成的关键酶基因,尽管目前从红花中克隆到了该基因,但在红花种子中未检测到亚麻酸,本研究旨在探究红花种子中不含亚麻酸的原因。利用红花种子转录组数据,根据FAD3基因的开放阅读框两端设计引物,分别以DNA和cDNA为模板进行克隆,并对其进行生物信息学分析、表达水平分析及功能分析。分析结果表明FAD3基因开放阅读框区不含内含子,FAD3氨基酸序列不存在信号肽,其第三个组氨酸保守区为HVVHH,而其他大多数植物均为HVIHH。FAD3蛋白相对保守,富含亮氨酸,存在3个组氨酸保守区。对CtFAD3氨基酸序列317位的缬氨酸替换成异亮氨酸后,发现FAD3基因仍不具有亚油酸脱氢酶活性,且红花中ω-3脂肪酸脱氢酶基因整体上在发育的种子中表达水平是极低的。可推测CtFAD3基因在种子中的低水平表达及其功能的丧失是导致红花种子中不含有α-亚麻酸的重要原因之一。