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A lot of textiles are unearthed from the Yingpan site. The climate of Yingpan is dry so the colors of the unearthed textiles are well preserved which provides reliable samples for dye identification. These textiles provide important evidence of cultural and technical exchange on the Silk Road during the Eastern Han and Jin Dynasties. It is of great significance to clarify the spread and development of textile dyes silk fiber raw materials and weaving techniques on the ancient Silk Road in China which not only enriches the information database of ancient textile cultural relics but also provides scientific basis for the subsequent protection restoration and display of cultural relics. Eight silk samples from the Yingpan site in Xinjiang were selected for study. On the one hand the dye and fiber species were identified. On the other hand the places of origin of these fabrics were analyzed according to the dye and fiber varieties and the characteristics of yarns and fabrics. High performance liquid chromatography-mass spectrometry HPLCMS was used to identify dye varieties and fiber micro-morphological analysis and proteomics methods were used to identify silk fiber varieties. By identifying the varieties of dyes and fibers of the eight samples and combining the identification results with the analysis of the structural characteristics of yarns and fabrics three major conclusions were drawn. Firstly there are mainly two types of dyes for silk fabrics unearthed from the Yingpan site in this study the main colorants detected in the red dye are alizarin and purpurin indicating that the source of the dye is Rubia tinctorum the presence of berberine as the main ingredient in the yellow dye with small amounts of palmatine and pharmacophorine suggests that the source is Phellodendron chinense. Rubia tinctorum is native in Xinjiang while Phellodendron chinense is grown in Sichuan Hunan and Hubei provinces. The sources of these two dyes exhibit distinct regional characteristics which provides important reference for place origin analysis. Secondly in this study the fiber materials of fabrics unearthed from the Yingpan site are all mulberry silk being either domestic silkworm Bombyx mori silk or wild mulberry silkworm Bombyx mandarina silk. The domestic silkworm silk is used for the simple single-layer textiles such as spun silk twill and patterned silk with simple weaves. The cross-section of the microscopic morphology of the used silk is a regular triangle and the longitudinal morphology of the silk fiber is straight which is consistent with the morphology of modern silk. The wild mulberry silkworm silk is used together with domesticated silkworm silk for more complex textiles such as Jin-tape and embroidery pillows. The cross-sectional shapes of silk fibers used for red yarns in Jin-tape 95BYYM14 9 and embroidered pillow 99BYYM2 5 are slightly rounded at one end and pointed at the other end which are consistent with the morphology of modern wild mulberry silk. The protein identification of the red yarn also reveals that it has the characteristic proteins of the wild mulberry silkworm silk. Both domestic silkworm silk and wild mulberry silkworm silk are used on Jin-tape and embroidery pillow which provides evidence for the use of wild mulberry silk in Xinjiang area and also reflects the richness and diversity of silk textiles unearthed in Xinjiang. Thirdly it is speculated that the red yarns of sample Jin-tape 95BYYM14 9 and embroidery pillow 99BYYM2 5 in this study are dyed with Rubia tinctorum in Xinjiang while the yellow yarns are made of domestic silkworm silk from Sichuan Hunan and Hubei and they are likely to be dyed with Phellodendron amurense locally and then imported into Xinjiang for weaving. This suggests that silk technology in the Han and Jin Dynasties was transmitted not only through finished products but also through semi-finished products accompanied by improvements in textile techniques. The article uses micronondestructive analysis test techniques to scientifically identify the dyes and fibers of some fabrics unearthed from the Yingpan site in Xinjiang and combines the characteristics of the yarns and fabrics from multiple angles to infer the place of weaving. It provides new evidence and support for the study of the transmission of textile materials technologies and culture along the Silk Road in Xinjiang and the research on the development of textile techniques. [ABSTRACT FROM AUTHOR]

Background and purpose: The osteogenic differentiation of MSCs plays a key role in bone tissue engineering and regenerative medicine, making it essential to explore natural compounds that may enhance this process. This aim this study was to investigate the osteogenic potential of both aqueous and alcoholic extracts derived from the root of Rubia tinctorum on rat bone marrow-derived mesenchymal stem cells (BM-MSCs). Materials and methods: Aqueous and alcoholic extracts were prepared from the root of Rubia tinctorum. Rat BM-MSCs were isolated and cultured and their osteogenic differentiation was assessed in the presence of varying concentrations of the extracts using alizarin red staining and alkaline phosphatase (ALP) activity. Cytotoxic effect of root extract of Rubia tinctorum was evaluated using MTT assay. Results: The MTT assay demonstrated that the root extract of Rubia tinctorum was cytotoxic at 1000 µg/ml. Additionally, the alizarin red assay showed that the most significant color intensity at 1 and 10 µg/ml concentrations on the 14th day. In terms of bone differentiation induction which measured via the alkaline phosphatase test, both the aqueous and alcoholic extracts at 1 and 10 µg/ml concentrations led to substantial increases (14.9- and 14.57-fold for the aqueous extract and approximately 7.37- and 7-fold for the alcoholic extract, respectively) in alkaline phosphatase activity compared to the negative control. Conclusion: This study sheds light on the osteogenic effects of Rubia tinctorum root extracts, emphasizing their potential as natural agents in promoting bone regeneration and tissue engineering applications. Further investigations into the underlying molecular mechanisms and in vivo studies are warranted to comprehensively evaluate the feasibility of utilizing these extracts for therapeutic bone regeneration purposes

This study focuses on the removal of methylene blue (MB), an organic dye, through adsorption on the Rubia tinctorum seeds (RTS) which are considered a new economical and efficient adsorbent for the removal of MB. This efficiency is proven by the microscopic analyses performed using Fourier-transform infrared spectroscopy, X-ray diffraction and scanning electron microscopy/energy-dispersive X-ray spectroscopy analysis techniques. These were performed before and after dye fixation and showed that the prepared surface of the RTS is made up of layers and pores, which could increase the total surface area and provide better fixation for the MB adsorbing cationic dye. To investigate the variation of the adsorption capacity, various parameters were tested such as the mass of the biosorbent, temperature, concentration, pH. The main parameters for maximum adsorption of MB were 40 min contact time and pH 5.00 and a percentage removal of 89.14% was obtained. Under optimal conditions, the monolayer adsorption capacity reached 188.68 mg/g. In addition, modeling studies showed that the adsorption process was best described by the pseudo-secondorder kinetic model and the isotherm of Langmuir. Finally, the thermodynamic study revealed the endothermic mode of the MB removal process by RTS. [ABSTRACT FROM AUTHOR]

Water is one of the greenest and most accessible solvents. To harness its potential, the water-based microwave-assisted extraction (MAE) of pigments from madder was optimized by the response surface methodology (RSM) using a Box–Behnken experimental design. The extract that presented both the highest extraction yield and the most intense color was obtained after a 30-s cycle at 1000 W using 100 mg of madder for 20 mL of water. This water-based MAE was more efficient than Soxhlet extraction and proved comparable to hydroalcoholic MAE. The optimized extract was further characterized using UHPLC-HRMS/MS to identify its main compounds. Glycosylated flavonoids and anthraquinones were putatively identified, as well as free anthraquinones, generally found in madder. The microwave-assisted extraction extended the range of polarity of the extracted compounds, making the water more versatile. [ABSTRACT FROM AUTHOR]

Plant cells are the biofactories of valuable chemicals. For efficient commercial production, in vitro biomass accumulation should be enhanced in the first place. For that purpose, growth regulators (auxins and cytokinins) at different concentrations have been frequently used in plant tissue cultures. Silver nanoparticles, which inhibit ethylene synthesis in plants, may be beneficial alternatives for growth regulators. Because of their practical and cost-effective production, biosynthetic silver nanoparticles were used in Rubia tinctorum node cultures in this study. Node explants were exposed to 0–30 mg L−1 biosynthetic silver nanoparticles in Woody Plant Medium (WPM) medium. At 10 mg L−1 silver nanoparticle concentration, shoot lengths and multiple shoot numbers were enhanced by 34% and 18%, respectively. In the same medium, shoot fresh and dry weights showed 52% and 22% increase, whereas 72% and 51% enhancements were recorded in root fresh and dry weights, respectively. At higher concentrations of silver nanoparticles (20 and 30 mg L−1), sharp drops were observed in growth parameters. Total phenolic, flavonoid, anthraquinone concentrations, total antioxidant capacity, radical scavenging activity, and chlorophyll and carotenoid contents of plantlet and root biomass were also examined. Except for total phenolic and flavonoid concentrations, all the other parameters were affected by the changing silver nanoparticle concentrations. The results showed that biosynthetic silver nanoparticles can be used to boost biomass and biochemical accumulations in R. tinctorum. The optimum dose of silver nanoparticles for better biomass accumulation was determined as 10 mg L−1. [ABSTRACT FROM AUTHOR]

Water is one of the greenest and most accessible solvents. To harness its potential, the water-based microwave-assisted extraction (MAE) of pigments from madder was optimized by the response surface methodology (RSM) using a Box–Behnken experimental design. The extract that presented both the highest extraction yield and the most intense color was obtained after a 30-s cycle at 1000 W using 100 mg of madder for 20 mL of water. This water-based MAE was more efficient than Soxhlet extraction and proved comparable to hydroalcoholic MAE. The optimized extract was further characterized using UHPLC-HRMS/MS to identify its main compounds. Glycosylated flavonoids and anthraquinones were putatively identified, as well as free anthraquinones, generally found in madder. The microwave-assisted extraction extended the range of polarity of the extracted compounds, making the water more versatile.

Abstract Objective Nanoharvesting from intact plants, organs, and cultured cells is a method in which nanoparticles are co-incubated with the target tissue, which leads to the internalization of nanoparticles. Internalized nanoparticles are coated in situ with specific metabolites that form a dynamic surface layer called a bio-corona. Our previous study showed that metabolites that form the bio-corona around anatase TiO2 nanoparticles incubated with leaves of the model plant Arabidopsis thaliana are enriched for flavonoids and lipids. The present study focused on the identification of metabolites isolated by nanoharvesting from two medicinal plants, Ocimum sanctum (Tulsi) and Rubia tinctorum (common madder). Results To identify metabolites that form the bio-corona, Tulsi leaves and madder roots were incubated with ultra-small anatase TiO2 nanoparticles, the coated nanoparticles were collected, and the adsorbed molecules were released from the nanoparticle surface and analyzed using an untargeted metabolomics approach. Similar to the results in which Arabidopsis tissue was used as a source of metabolites, TiO2 nanoparticle bio-coronas from Tulsi and madder were enriched for flavonoids and lipids, suggesting that nanoharvesting has a wide-range application potential. The third group of metabolites enriched in bio-coronas isolated from both plants were small peptides with C-terminal arginine and lysine residues.

Objective(s): Nowadays, nanotechnology has offered great success in resolving concerns in cancer therapy and created a new interdisciplinary field of study incorporating various sciences, such as biology, chemistry and medicine. Apoptosis is a conserved and controlled strategy in regulating cellular growth and proliferation, as well as preserving development and general homeostasis of the body. Zinc oxide nanoparticles (ZnO-NPs) are the most important and widely used nanoparticles. This study aimed to evaluate the apoptosis-inducing properties of the synthesized ZnO-NPs by aqueous extract of Rubia tinctorum against the MCF7 breast cancer cell line. Materials and Methods: Zinc oxide nanoparticles were synthesized using Rubia tinctorum extract and characterized by some methods including dynamic light scattering (DLS), field emission scanning electron microscopy (FESEM) and x-ray diffraction analysis (XRD). Apoptosis was measured by the Hoechst and Acridine-Orange/Propodium Iodide staining, as well as flow cytometry. Results: The results of this study showed that the particle size of biosynthesized ZnO-NPs using R.tinctorum extract was about 40 nm and had a spherical morphology. The obtain results of the Hoechst and Acridine-Orange/Propodium Iodide staining, as well as flow cytometry showed that biosynthesized ZnO-NPs effectively and dose-dependently induced apoptosis in the MCF7 breast cancer cells.Conclusion: Therefore, the biosynthesized ZnO-NPs by watery extract of R. tinctorum can be used in the treatment of many diseases, including cancers.

Diseases caused by various drug-resistant strains in plants are increasing in many countries of the world, so many efforts have been made to find new compounds as a suitable alternative to chemical drugs and pesticides. In this study, the antimicrobial effect of essential oils and ethanolic extracts of 10 medicinal plants were investigated on Rathayibacter tritici and Xanthomonas translucens. Alcoholic extracts of medicinal plants were extracted using a rotary apparatus. Two standard bacteria R. tritici and X. translucens were prepared from Persian Type Culture Collection. The minimum inhibitory concentration and the minimum inhibitory concentration of essential oils and ethanolic extracts of plants used at a concentration of 50 mg/ ml were determined by dilution in liquid medium on pathogens. Based on the results, the lowest inhibitory concentration of thyme essential oil was 6.25 ppm, which was inhibited by R. tritici, and the lowest concentration of Hypericum perforatum essential oil against X. translucens was 6.25 ppm. The lowest concentrations of essential oils of yew and fennel were 6.25 ppm, which were inhibited by both bacteria. Rubia tinctorum leaf essential oil in a concentration of 6.25 only inhibited R. tritici bacteria. The antibacterial properties of the essential oils of the studied plants were higher than the extract. Essential oils of yew and oleander were the most effective against R. tritici and X. translucens, followed by thyme and rosemary against R. tritici and herring flower against X. translucens . Although the clinical use of ethanolic extracts and essential oils of the studied plants seems valuable due to side effects, but for the clinical use of essential oils and extracts, more research should be done on the mechanism of action of effective compounds of these plants on microbial agents. [ABSTRACT FROM AUTHOR]

Plants have gained great importance. Secondary metabolites contribute to the drug discovery and development by their bioactive properties. Rubia tinctorum L. essential oil (EO) was obtained and analysed. Antioxidant and antibacterial activities were evaluated. The plant's EOs were obtained through steam distillation, and the compounds were identified using gas chromatography-mass spectrometry (GC-MS) analysis. DPPH free radical scavenging and ferric-reducing antioxidant power (FRAP) were employed to assess antioxidant activity. Total antioxidant capacity (TAC) was also presented. The disc diffusion method was employed for testing antibacterial activity. Cyclohexanone was identified as the predominant component in the EO, constituting 88.74% of the total composition. The EO did not show significant antioxidant capacity, while it demonstrated antimicrobial effect against Bacillus cereus ATCC 6633 (>13 mm of inhibition; 500 mg/mL) and Shigella ATCC 12022 (≥12 mm of inhibition; 500 mg/mL). R. tinctorum L. is new source of cyclohexanone.

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The roots of the madder plant (Rubinia tinctorum) contain some pigments that can be used to produce the red dyes. In order to study the effect of salinity on alterations in the amount of plant pigment an experiment was conducted according to a completely randomized design with three replications. Based on the CIE XYZ color space model designed close to human color vision, the results showed that increasing soil salinity decreased the value of The lightness value parameter (L*), but the A* (redness and greenness) and B* (yellowness and blueness) values were increased with increasing soil salinity up to the level of 8 dSm-1. There was a significant positive correlation between A* value and Ca + Mg and a significant negative correlation between L* value and Ca+Mg at the 5% level. Fitting three nonlinear regression model to data, simulated and described the variation of colorimetric specification of woolen samples dyed under different madder dyes and salinity conditions. These models proved to be an appropriate and reasonable link between madder growth conditions and colorimetric specification of woolen samples dyed such as L*, A* and B* values. Due to the high resistant characteristics of madder to salinity, increasing salinity in the madder root zone lead to acceptable quality and efficiency of pigment production and to expand the possibilities for economic production of Rubinia tinctorum in soil salinity in the central arid region of Iran

Madder (Rubia tinctorum L.) is a perennial plant that its roots and rhizomes have rich anthraquinone (AQ) derivatives including alizarin and purpurin. This study was carried out to determine the effects of salicylic acid (SA) and L-phenylalanine (L-Phe) applications on the root growth parameters and secondary metabolite accumulation in madder adventitious roots derived from internode parts without needing to be collected from nature. For this aim, two different L-Phe (50 and 100 µM) and SA (20 and 40 µM) were added separately and together to the liquid Murashige and Skoog (MS) medium cultured adventitious roots for 7 days. Then roots were evaluated in terms of fresh root weight, root growth index, dry root weight, and contents of total AQ, alizarin, purpurin, total phenolic contents (TPC), and some important phenolic compounds. According to the results, L-Phe stimulated the root growth of madder while the effects of SA on root growth parameters varied depending upon its concentrations. L-Phe had no significant influence on the total AQ, alizarin, and purpurin. Conversely, SA increased the AQs, and 20 µM SA was the most suitable application providing the greatest total AQ, alizarin, and purpurin. TPC and individual phenolic compounds changed according to the applications. Not only L-Phe but also SA had positive effects on the phenolic accumulation in adventitious roots. It was determined that the combinations of 40 µM SA and 100 µM L-Phe were the most effective applications in terms of phenolic accumulation. Key message: Madder is a plant whose popularity has increased in food, cosmetics, pharmacy and recently medicine. This research was designed in order to produce high amounts of important secondary metabolites in adventitious root cultures of madder without the need to collect plants from nature. In this study, it has been shown that L-phenylalanine and salicylic acid applied to adventitious roots remarkably increase the production of anthraquinones and phenolics when used in appropriate concentrations and combinations. [ABSTRACT FROM AUTHOR]

Objective: Nanoharvesting from intact plants, organs, and cultured cells is a method in which nanoparticles are co-incubated with the target tissue, which leads to the internalization of nanoparticles. Internalized nanoparticles are coated in situ with specific metabolites that form a dynamic surface layer called a bio-corona. Our previous study showed that metabolites that form the bio-corona around anatase TiO2 nanoparticles incubated with leaves of the model plant Arabidopsis thaliana are enriched for flavonoids and lipids. The present study focused on the identification of metabolites isolated by nanoharvesting from two medicinal plants, Ocimum sanctum (Tulsi) and Rubia tinctorum (common madder). Results: To identify metabolites that form the bio-corona, Tulsi leaves and madder roots were incubated with ultra-small anatase TiO2 nanoparticles, the coated nanoparticles were collected, and the adsorbed molecules were released from the nanoparticle surface and analyzed using an untargeted metabolomics approach. Similar to the results in which Arabidopsis tissue was used as a source of metabolites, TiO2 nanoparticle bio-coronas from Tulsi and madder were enriched for flavonoids and lipids, suggesting that nanoharvesting has a wide-range application potential. The third group of metabolites enriched in bio-coronas isolated from both plants were small peptides with C-terminal arginine and lysine residues. [ABSTRACT FROM AUTHOR]

Objective(s): Nowadays, nanotechnology has offered great success in resolving concerns in cancer therapy and created a new interdisciplinary field of study incorporating various sciences, such as biology, chemistry and medicine. Apoptosis is a conserved and controlled strategy in regulating cellular growth and proliferation, as well as preserving development and general homeostasis of the body. Zinc oxide nanoparticles (ZnO-NPs) are the most important and widely used nanoparticles. This study aimed to evaluate the apoptosis-inducing properties of the synthesized ZnO-NPs by aqueous extract of Rubia tinctorum against the MCF7 breast cancer cell line. Materials and Methods: Zinc oxide nanoparticles were synthesized using rubia tinctorum extract and characterized by some methods including dynamic light scattering (DLS), field emission scanning electron microscopy (FESEM) and x-ray diffraction analysis (XRD). Apoptosis was measured by the Hoechst and Acridine-Orange/Propodium Iodide staining, as well as flow cytometry. Results: The results of this study showed that the particle size of biosynthesized ZnO-NPs using R.tinctorum extract was about 40 nm and had a spherical morphology. The obtain results of the Hoechst and Acridine- Orange/Propodium Iodide staining, as well as flow cytometry showed that biosynthesized ZnO-NPs effectively and dose-dependently induced apoptosis in the MCF7 breast cancer cells. Conclusion: Therefore, the biosynthesized ZnO-NPs by watery extract of R. tinctorum can be used in the treatment of many diseases, including cancers. [ABSTRACT FROM AUTHOR]

The use of nanocarriers (NCs), i.e., nanomaterials capable of encapsulating drugs and releasing them selectively, is an emerging field in agriculture. In this study, the synthesis, characterization, and in vitro and in vivo testing of biodegradable NCs loaded with natural bioactive products was investigated for the control of certain phytopathogens responsible for wood degradation. In particular, NCs based on methacrylated lignin and chitosan oligomers, loaded with extracts from Rubia tinctorum, Silybum marianum, Equisetum arvense, and Urtica dioica, were first assayed in vitro against Neofusicoccum parvum, an aggressive fungus that causes cankers and diebacks in numerous woody hosts around the world. The in vitro antimicrobial activity of the most effective treatment was further explored against another fungal pathogen and two bacteria related to trunk diseases: Diplodia seriata, Xylophilus ampelinus, and Pseudomonas syringae pv. syringae, respectively. Subsequently, it was evaluated in field conditions, in which it was applied by endotherapy for the control of grapevine trunk diseases. In the in vitro mycelial growth inhibition tests, the NCs loaded with R. tinctorum resulted in EC90 concentrations of 65.8 and 91.0 μg·mL−1 against N. parvum and D. seriata, respectively. Concerning their antibacterial activity, a minimum inhibitory concentration of 37.5 μg·mL−1 was obtained for this treatment against both phytopathogens. Upon application via endotherapy on 20-year-old grapevines with clear esca and Botryosphaeria decay symptoms, no phytotoxicity effects were observed (according to SPAD and chlorophyll fluorescence measurements) and the sugar content of the grape juice was not affected either. Nonetheless, the treatment led to a noticeable decrease in foliar symptoms as well as a higher yield in the treated arms as compared to the control arms (3177 vs. 1932 g/arm), suggestive of high efficacy. Given the advantages in terms of controlled release and antimicrobial product savings, these biodegradable NCs loaded with natural extracts may deserve further research in large-scale field tests.

Madder (Rubia tinctorum L.) is a valuable plant rich in anthraquinones having dying properties and biological activities. This study was carried out to determine the effect of sodium chloride (NaCl) applications on the root growth and secondary metabolite accumulation in adventitious roots of madder. For this aim, adventitious roots derived from stem explants in in vitro conditions were cultured in MS medium containing different concentrations of NaCl (0, 1, 2, 3 and 4 g/l) for 7 days. Then roots were evaluated in terms of root growth index, total AQ, alizarin, purpurin and total phenolic contents. Based on the results, root growth decreased in line with the elevating level of NaCl while secondary metabolite accumulation significantly increased with NaCl applications compared to the controls. It was determined that NaCl at 3 g/l concentration was the most effective application in terms of total AQ, alizarin, purpurin and phenolic accumulation.

Abstract Background: Angiogenesis occurs in physiologic (wound healing) and pathological conditions and plays an important role in tumor growth, progression and metastasis. Therefore, the inhibition of angiogenesis can be an important approach to treat cancer diseases. In recent years, the use of nanoparticles has been developed to control cancer cells and prevent tumor growth. Various studies have shown the role of nanoparticles in reducing angiogenesis and inhibition of cancer cells. So, in this study, the anti-angiogenic effect of silver nanoparticle synthesis by Rubina tinctorum L (Ru-AgNPs) on Chicken Chorioallantoic Membrane (CAM) was investigated. Materials and Methods: In an experimental study, 60 eggs were randomly divided into 6 groups including control, laboratory control and 4 treatment groups (10 eggs/group). On the second day of incubation, a window was created on the eggs. On the 8th day, the chorioallantoic membrane of samples was treated with different concentrations of nanoparticles and on the 12th day, the image was taken from all samples. Finally, the number and length of vessels on the chorioallantoic membrane and following that weight and length of the embryo from crown to squattiest were measured. Quantitative data were analyzed using LSD-test. Results: The results showed that the Ru-AgNPs significantly reduced the length and number of blood vessels in the treated groups compared to the control group. Also, the comparison of length and weight of embryos in treated samples with control group showed that the Ru-AgNPs had an inhibitory effect on the length of the embryos, but the weight did not differ significantly in the treatment with the control group. Conclusion: Considering the inhibitory effect of the Ru-AgNPs on the angiogenesis of the chorioallantoic membrane, with further studies, this compound can be used to control and inhibit pathological angiogenesis in many diseases such as cancers.

Madder is one of the most important crops that used for medical and industrial applications and is widely cultivated in Yazd province. During 2012, sampling was done form seeds madder in important areas planted in Yazd province, including Bafq and Ardakan. After culturing and purification of fungal isolates in PDA and CLA media, additional identification was performed by PCR with specific primers for each species. Detection of fungi mycotoxins producing potential such as Nivalenol (NIV) using Tri13 primers was done. High-performance liquid chromatography (HPLC) was used to confirm the produce NIV mycotoxins potential in Fusarium species. 249 fungal strains were isolated from madder seed belonging to 6 genera of fungi including Fusarium spp., Aspergillus spp., Penicillium spp., Alternaria spp., Rhizoctonia solani and Rhizpous spp., that Fusarium isolates with 71 percent was the most frequency among fungi isolated. Among Fusarium fungi isolated, F. solani (55 isolates) and F. oxysporum (41 isolates) were the most frequency. F. poae, F. semitectum and F. equiseti ability to produce mycotoxins such as Nivalenol (NIV) that are harmful to human health and animals as well as effect on the quantity and quality of madder color production. Tri13 gene involved in production NIV was detected in three Fusarium species that all isolates produce NIV. The results of HPLC showed that all studied Fusarium fungi, have the potential to produce NIV mycotoxins. The results of this study showed that fungi associated with seeds madder are able to produce trichothecene mycotoxins that they can be dangerous for consumers. Given that, this is the first report of fungi mycotoxins producing on seeds madder in Yazd province, thus should be measures to control and reduce fungal agents in these products.

Salinity which covers vast areas of the world is increasing every year. But some plants like madder can grow in these areas. Madder (Rubia tinctorum) is a perennial plant species from the Rubiaceae family. In this study, madder plants were first treated by different concentration of NaCl (100, 200, 300, and 400 mM). Then gene expression of salinity stress was studied. For gene study, vacuolar H+-pyrophosphatase pump (AVP) and tonoplast Na+/H+ antiporters (NHX) from madder plant were isolated and sequenced. Analyzing protein sequences of these genes demonstrated that the protein sequences have high similarity with the same genes in other plants. Constructing phylogenetic trees based on the protein sequences of the AVP and NHX genes, we found high similarity with Coffea arabica and Capsicum annuum, respectively. Studying gene expression of the AVP and NHX under the condition of salt stress revealed that the genes were up-regulated, which continues up to 400 mM of salt concentration. [ABSTRACT FROM AUTHOR]

The roots of the madder plant (Rubinia tinctorum) contain some pigments that can be used to produce the red dyes. In order to study the effect of salinity on alterations in the amount of plant pigment an experiment was conducted according to a completely randomized design with three replications. Based on the CIE XYZ color space model designed close to human color vision, the results showed that increasing soil salinity decreased the value of The lightness value parameter (L*), but the A* (redness and greenness) and B* (yellowness and blueness) values were increased with increasing soil salinity up to the level of 8 dSm-1. There was a significant positive correlation between A* value and Ca + Mg and a significant negative correlation between L* value and Ca+Mg at the 5% level. Fitting three nonlinear regression model to data, simulated and described the variation of colorimetric specification of woolen samples dyed under different madder dyes and salinity conditions. These models proved to be an appropriate and reasonable link between madder growth conditions and colorimetric specification of woolen samples dyed such as L*, A* and B* values. Due to the high resistant characteristics of madder to salinity, increasing salinity in the madder root zone lead to acceptable quality and efficiency of pigment production and to expand the possibilities for economic production of Rubinia tinctorum in soil salinity in the central arid region of Iran. [ABSTRACT FROM AUTHOR]

In the past decades, Food colorants which are used in food industries are suspected due to the increased world sensitivity against the health of human diet; hence, many of researchers tried to extract the edible colorants from natural resources. One of the natural resources is the root of madder (Rubia tinctorum) which has the most stable natural red pigment, Alizarin. In this research the madder extract as natural colorant at 2, 3 and 4% and pectin as a replacer for gelatin at 3:0, 1.5:1.5 and 0:3% were used in flavoured milk- based dessert and their various qualitative parameters were evaluated during 15 days of storage. The results showed that increasing of madder extract with various ratios of gelatin: pectin, changed the dry matter and pH of the specimens significantly (p<0.05) and after 15 days, the pH raised up clearly. The texture evaluation showed that in the samples containing 3:0 gelatin: pectin, were more rigid than the others and demonstrated the least textural changes during 15 days according to the lower syneresis (p<0.05). The results of sensory evaluation showed that the addition of madder extract at its maximum level (4%), had the best results about the color, in the viewpoint of panelists and had no side effects on other parameters like odor, taste, apparent shape, texture and overall acceptance. Totally and according to the results, it can be concluded that by addition of 3% of gelatin along with 4% of madder extract in flavoured milk- based dessert, the qualitative parameters can be improve along with more shelf life and marketability, too. [ABSTRACT FROM AUTHOR]

The aim of the present study was to establish a mild extraction and universal method to characterize madder lakes of Rubia tinctorum by liquid chromatography coupled with a photodiode array detector. To analyze the lakes, anthraquinone molecules must be decomplexed of metal links. To this end, two processes, one ultrasound and the other microwave, were improved in association with two solutions, an oxalic acid solution (0.5 M in MeOH/H 2 O 50/50) and an acetic acid buffer solution (1 M). First, the decomplexation of an alizarin experimental lake was optimized in comparison with a reference method using a strong acid. The microwave process used oxalic acid and increased the decomplexation yield of alizarin (71%) compared with the reference method (31%). Second, different madder experimental lakes, which were prepared using different metal salts, were decomplexed. The obtained results suggest that the use of microwave associated with oxalic acid solution is the most universal method providing a decomplexation of anthraquinones from lakes without hydrolysis of glycosidic compounds occurring. • Synthesis of madder lakes using five different metallic cations. • Decomplexation of metal-complex dyes by two separate processes: ultrasound and microwave. • Development of a universal, efficient, and nondenaturing method for the study of madder lakes. [ABSTRACT FROM AUTHOR]

Cancer is a devastating disease and is considered number one killer worldwide. Herbal formulations had played a key role over the past several decades in the development of anti-cancer drugs. Medicinal plants, which are endemic in Jordan, are known for several biological activities in particular their anti-cancer activity. However, the anti-cancer efficacy of the root extracts of Jordanian Rubia tinctorum and Alkanna tinctoria is not yet reported. To address this issue, this study assessed the anti-cancer activity of some root extracts obtained from Jordanian R. tinctorum and A. tinctoria in different tumor cell lines including the tongue, bladder, colon, gastric, lungs, breast, pancreas, and renal tissue origins by modified propidium iodide (PI) based monolayer assay. Among the tested root extracts obtained by different solvent systems, A. tinctoria in 100 % ethanol and methanol showed prominent anti-cancer activity against MDA-MB-231breast cancer cells (IC50: 2.98 μg/ml, IC70: 6.03 μg/ml), and CAL-27 tongue squamous carcinoma cells (IC50: 3.86 μg/ml, IC70: 5.97 μg/ml) respectively. Different solvent root extracts of R. tinctorum exhibited a similar trend of anti-tumor activity in both CAL-27 and MDAMB- 231 cells. The anti-proliferative property of the extracts on CAL-27 and MDA-MB-231 cells is unclear. However, it can be concluded that the observed anti-cancer potential can be attributed to the phenolic compounds of the extracts as high polar solvents were used for extraction. The current study forms the rationale for isolating significant amount of anti-cancer active compounds from R. tinctorum and A. tinctoria. [ABSTRACT FROM AUTHOR]

[Display omitted] • Rubia Tinctorum for efficient adsorption of CV dye from aqueous solutions. • The maximum absorption capacity was found to be 174 mg/g. • The as-developed composite provides superior cleanability and selectivity for the cationic dye Crystal Violet Water. • The adsorption mechanism includes hydrogen bonding and electrostatic interactions. • The composite were an effective and reusable biosorbent for cleaning CV contaminated effluents. The current study focuses on the adsorption of crystal violet (CV), a cationic organic dye, employing Rubia Tinctorum (RTS) seeds, a novel class of economically viable and effective adsorbents that can bind CV. Thereafter, X-ray diffraction (XRD), scanning electron microscopy (SEM), and a Fourier transform infrared spectrometer were used to characterize it (FT-IR). The polymer was used in conjunction with the removal of dye such as cellulose, which is the molecule's natural carrier. As a result, the central composite surface response (RSM-CCD) approach was used to analyze and optimize the impacts of four factors on removal effectiveness, including pH, adsorbent quantity, contact duration, and beginning dye concentration. The following settings were found to be the best for maximal adsorption: 10.0 mg RTS, 30 min of contact duration, and pH 6.00. A percentage reduction of 88.23 % was obtained for CV under ideal circumstances. 174 mg/g was the monolayer adsorption capability. The hydrophobic interactions, hydrogen bonds, n-electron donor/acceptor interactions and electrostatic interactions. Theoretical DFT simulations are performed to better understand the atomic interactions that exist between the CV molecule and the RTS surface. According to our study of the adsorption kinetics with the effect of the pollutant concentration, we found that the Langmuir model and the pseudo-second-order model are the most appropriate to explain the adsorption phenomena, with the highest correlation rate. The high value of E HOMO indicates that the CV molecule is ready to accept electron transfer, which favors its adsorption to the surface of the polymer used in conjunction with the elimination of the CV dye, which is the cellulose affecting electron transfer at the adsorbed fibers. Here again, simulation calculations confirm our experience and the rubia tinctorum plant can be used as a possible biosorbent material for the elimination of crystal violet dye from an aqueous solution. [ABSTRACT FROM AUTHOR]

Background: Although carmine, as a synthetic dye, is the major substance for staining helminths, it may impose some adverse effects on human health. In the present study, we evaluated the aqueous extracts of onion (Allium cepa) skin, walnut (Juglans regia) husk, and madder (Rubia tinctorum) roots as potential herbal dyes for staining parasitic helminths. Methods: Aqueous solutions (5%, 10% and 20%, w/v) of each herbal prepared from dried and powdered husk of walnut, skins of red onion, and madder roots in distilled water. Parasitic helminths including Fasciola spp., Dicrocoelium denderiticum, Echinococcus gronulosus protoscolices, Moniezia spp., and Haemonchus contortus were stained by different concentrations of herbal dyes according to carmine staining method. The structural clarity and quality of stained internal organs of the helminths such as suckers, intestine, and reproductive systems were scored by semi-quantitative evaluation in comparison with carmine stained samples. Results: The optimum concentrations of extracts for helminths staining were 10% (w/v) of A. cepa and J. regia, and 20% (w/v) of R. tinctorum with final scores of 3.1, 3 and 2.8, respectively. In general, A. cepa and J. regia extracts showed higher quality in staining Platyhelminthes, while R. tinctorum extract presented relatively higher quality in staining Nematoda. Conclusion: Considering proper quality of A. cepa, J. regia and R. tinctorum extracts in staining the helminths, they may be safe, eco-friendly, and inexpensive alternatives to carmine dye.

The complex mixtures of colorants present in different madder species can provide significant information about which plant species or technique was used to dye the fibres of historical textile artefacts, hence, when extracting and analysing colorants from textile artefacts as much of this information as possible should be preserved. Historical textiles are most commonly extracted with 37% hydrochloric acid: methanol: water (2:1:1, v/v/v ), but this solvent system hydrolyses dye glycosides and may also induce chemical reactions. One of the primary components in Dyers' madder ( Rubia tinctorum L.) is lucidin primeveroside, but it is rarely seen in artefacts, nor is the corresponding aglycon lucidin. It has been demonstrated that the hydrochloric acid method causes hydrolysis of anthraquinone glycosides to their aglycon counterpart. Herein it is demonstrated that lucidin is not stable in such acidic conditions and degrades rapidly to xanthopurpurin. This is confirmed by HPLC, LC-MS and 1 H NMR, which also provide evidence of the mechanism of degradation being a retro-aldol process. [ABSTRACT FROM AUTHOR]

Background: Angiogenesis occurs in physiologic (wound healing) and pathological conditions and plays an important role in tumor growth, progression and metastasis. Therefore, the inhibition of angiogenesis can be an important approach to treat cancer diseases. In recent years, the use of nanoparticles has been developed to control cancer cells and prevent tumor growth. Various studies have shown the role of nanoparticles in reducing angiogenesis and inhibition of cancer cells. So, in this study, the anti-angiogenic effect of silver nanoparticle synthesis by Rubina tinctorum L (Ru-AgNPs) on Chicken Chorioallantoic Membrane (CAM) was investigated. Materials and Methods: In an experimental study, 60 eggs were randomly divided into 6 groups including control, laboratory control and 4 treatment groups (10 eggs/group). On the second day of incubation, a window was created on the eggs. On the 8th day, the chorioallantoic membrane of samples was treated with different concentrations of nanoparticles and on the 12th day, the image was taken from all samples. Finally, the number and length of vessels on the chorioallantoic membrane and following that weight and length of the embryo from crown to squattiest were measured. Quantitative data were analyzed using LSD-test. Results: The results showed that the Ru-AgNPs significantly reduced the length and number of blood vessels in the treated groups compared to the control group. Also, the comparison of length and weight of embryos in treated samples with control group showed that the Ru-AgNPs had an inhibitory effect on the length of the embryos, but the weight did not differ significantly in the treatment with the control group. Conclusion: Considering the inhibitory effect of the Ru-AgNPs on the angiogenesis of the chorioallantoic membrane, with further studies, this compound can be used to control and inhibit pathological angiogenesis in many diseases such as cancers. [ABSTRACT FROM AUTHOR]

Background: Although carmine, as a synthetic dye, is the major substance for staining helminths, it may impose some adverse effects on human health. In the present study, we evaluated the aqueous extracts of onion (Allium cepa) skin, walnut (Juglans regia) husk, and madder (Rubia tinctorum) roots as potential herbal dyes for staining parasitic helminths. Methods: Aqueous solutions (5%, 10% and 20%, w/v) of each herbal prepared from dried and powdered husk of walnut, skins of red onion, and madder roots in distilled water. Parasitic helminths including Fasciola spp., Dicrocoelium denderiticum, Echinococcus gronulosus protoscolices, Moniezia spp., and Haemonchus contortus were stained by different concentrations of herbal dyes according to carmine staining method. The structural clarity and quality of stained internal organs of the helminths such as suckers, intestine, and reproductive systems were scored by semiquantitative evaluation in comparison with carmine stained samples. Results: The optimum concentrations of extracts for helminths staining were 10% (w/v) of A. cepa and J. regia, and 20% (w/v) of R. tinctorum with final scores of 3.1, 3 and 2.8, respectively. In general, A. cepa and J. regia extracts showed higher quality in staining Platyhelminthes, while R. tinctorum extract presented relatively higher quality in staining Nematoda. Conclusion: Considering proper quality of A. cepa, J. regia and R. tinctorum extracts in staining the helminths, they may be safe, eco-friendly, and inexpensive alternatives to carmine dye. [ABSTRACT FROM AUTHOR]

Background: Inflammation is a dynamic process that is elicited in response to various harmful stimulantsthat may threaten the well-being of the host. Herbs have been traditionally used as anti-inflammatory agents since very early times. Rubia tinctorum L. is one of these herbs that havebeen used for treating inflammatory diseases in Iranian traditional medicine. Objective: This study aims at considering anti-inflammatory effects of R. tinctorum and comparing its extracts in this property. Methods: The root of Rubia tinctorum (Rubiaceae) was collected from Yazd province, in the center of Iran. The root extracts of plant were studied for their anti-inflammatory activity by using carrageenan-induced hind paw edema in Wistarmale rats. Indomethacin and normal saline were used as positive and negative controls, respectively. Different concentrations of aqueous, ethanolic, and 50% aq. ethanolic extracts (600, 800, 1000 mg/kg) were given orally before carrageenan injection. The paw thickness was measured at 1, 3 and 5 hours after injection. Results: Both, aqueous and 50% eq. ethanolic extracts reduced paw edema at 600 and 800 mg/kg concentrations. Moreover, ethanolic extract showed significant effect only in1sthour after carrageenan injection. Conclusion: The results indicated that the aqueous extract were exhibited higher anti-inflammatory effect compared with other extracts. Accordingly, the potent anti-inflammatory effect of the root extracts is attributed to the polar compounds which are extracted in the water.

Anthraquinones (AQs) are secondary metabolites widely distributed in nature. Interesting applications of plant extracts containing AQs include the treatment of Hepatitis C and cancer. Plant in vitro culture is an alternative for producing plant-derived pharmaceuticals in controlled conditions and with low environmental impact. Moreover, it allows the application of different strategies for enhancing secondary metabolite production. Hairy root cultures, obtained after Agrobacterium rhizogenes infection, are able to produce high amounts of secondary metabolites at high growth rates. In this work, growth kinetics of hairy root cultures of Rubia tinctorum and AQ production were evaluated in two different culture media, Gamborg B5 with half of the saline strength (B5 1/2 ) and Lloyd & Mc Cown’s Woody Plant Medium (WPM). Although WPM allowed higher biomass production (58.6% higher) than B5 1/2 , specific AQ production was higher in B5 1/2 (between 1.2 and 2.1 fold increases from day 21 to the end of the experiment). Moreover, AQ release to the culture medium was observed in B5 1/2 (∼10% of total AQs). The different performance of hairy roots in these culture media may be due to a limiting nutrient (other than carbon source) in B5 1/2 . Elicitation in B5 1/2 with methyl jasmonate (100 μM) resulted in a massive accumulation of intracellular (between 1.5 and 2.4-fold increases) and also extracellular AQs (up to 8.1 fold-increase compared with control at 4 days post-elicitation), which could ease AQ purification. These results prove the usefulness of combining different approaches to enhance secondary metabolite accumulation in plant in vitro cultures, in order to develop an optimized productive process. [ABSTRACT FROM AUTHOR]

This study was carried out to determine the effects of methyl jasmonate (MeJA) and caffeic acid (CA) applications on the root growth index and contents of total anthraquinones (AQs), alizarin, purpurin, total phenolic (TP) and individual phenolic compounds in madder (Rubia tinctorum) roots grown in vitro conditions. For this aim, in vitro adventitious roots derived from internode parts were cultured in Murashige and Skoog media containing two different concentrations of MeJA (10 and 100 µM) and CA (1 and 2 mM) for 7 days. Based on the results, MeJA decreased the root growth index compared to control while CA had a positive influence on the root growth of madder. It may be concluded that 2 mM CA+100 µM MeJA combination was the most suitable application providing the highest AQs and phenolics. [ABSTRACT FROM AUTHOR]

Madder ( Rubia tinctorum L.) has been widely used as a red dye throughout history. Acid-sensitive colorants present in madder, such as glycosides (lucidin primeveroside, ruberythric acid, galiosin) and sensitive aglycons (lucidin), are degraded in the textile back extraction process; in previous literature these sensitive molecules are either absent or present in only low concentrations due to the use of acid in typical textile back extraction processes. Anthraquinone aglycons alizarin and purpurin are usually identified in analysis following harsh back extraction methods, such those using solvent mixtures with concentrated hydrochloric acid at high temperatures. Use of softer extraction techniques potentially allows for dye components present in madder to be extracted without degradation, which can potentially provide more information about the original dye profile, which varies significantly between madder varieties, species and dyeing technique. Herein, a softer extraction method involving aqueous glucose solution was developed and compared to other back extraction techniques on wool dyed with root extract from different varieties of Rubia tinctorum . Efficiencies of the extraction methods were analysed by HPLC coupled with diode array detection. Acidic literature methods were evaluated and they generally caused hydrolysis and degradation of the dye components, with alizarin, lucidin, and purpurin being the main compounds extracted. In contrast, extraction in aqueous glucose solution provides a highly effective method for extraction of madder dyed wool and is shown to efficiently extract lucidin primeveroside and ruberythric acid without causing hydrolysis and also extract aglycons that are present due to hydrolysis during processing of the plant material. Glucose solution is a favourable extraction medium due to its ability to form extensive hydrogen bonding with glycosides present in madder, and displace them from the fibre. This new glucose method offers an efficient process that preserves these sensitive molecules and is a step-change in analysis of madder dyed textiles as it can provide further information about historical dye preparation and dyeing processes that current methods cannot. The method also efficiently extracts glycosides in artificially aged samples, making it applicable for museum textile artefacts. [ABSTRACT FROM AUTHOR]

Lithium pollution may seriously influence the metabolic and signalling processes of plants. In the present paper, we investigate the effect of lithium chloride on fungal elicitor-triggered H2O2 generation in Rubia tinctorum L. cell cultures. Our results show that Li+ strongly influences elicitor-induced H2O2 formation and time-course in the cells nad culture medium. Neomycin, a phospholipase C inhibitor, and 2-APB, an inositol-1,4,5-triphosphate (IP3) receptormediated Ca2+ release blocker, strongly affected the elicitor-induced H2O2 production and had a similar effect on elicitor-triggered H2O2 formation as Li+. We monitored changes in H2O2 location at subcellular level and our observations confirmed the changes measured by quantitative methods. The obtained results enabled us to deduce that the IP3 pathway might be involved in the early signalling events leading to the moderation of elicitor-induced reactive oxygen species generation.

Purpurin (1,2,4-trihidroxyanthraquinone), a naurally occuring anthraquinone pigment present in the root of Rubia tinctorum (Mull. Arg.) Zahlbr. was isolated from the plant, purified and characterized by HPLC chromatography, UV-Vis and NMR spectroscopy. The geometries of the purpurin conformers were optimized using the B3LYP/6-311+G(d,p) level of theory. It was found that all conformations have similar energies and that the corresponding purpurin radicals are planar. In spite of their planarity, there is no significant electron delocalization over the A and C rings. The antioxidant properties of purpurin were investigated using the colorimetric assay as Trolox-equivalent antioxidant capacity, and theoretical BDE (Bond Dissociation Enthalpy). Both methods revealed that purpurin has strong antioxidant capacity.