Crocin Content Research Articles

Quercetin and kaempferol from saffron petals alleviated hydrogen peroxide-induced oxidative damage in B16 cells.

Saffron petals are usually considered as waste after saffron harvest. However, saffron petals contain many important phytochemical components (e.g. quercetin and kaempferol), which may alleviate oxidative damage in human cells. The contents of flavonoids and crocin in different parts of saffron were analyzed. The protective effects of flavonoids from saffron on oxidative damage of B16 cells were investigated. Saffron stigma contained high contents of crocin and picrocrocin, whereas flavonoid content (quercetin, 4.03 ± 0.33 mg g-1 DW; kaempferol, 47.80 ± 0.60 mg g-1 DW) was higher in saffron petals than in other parts. Incubation of B16 cells with quercetin (10-30 μmol L-1) and kaempferol (20-30 μmol L-1) obtained from saffron extracts could significantly increase the total antioxidant capacity (T-AOC) and the activity of NADPH:dehydrogenase quinone-1 (NQO1) to alleviate H2O2-induced oxidative damage. Quercetin was better than kaempferol in increasing NQO1 activity and T-AOC. Quercetin extracted from saffron petals could induce NQO1 expression through regulating kelch-like ECH-associated protein-1/nuclear factor erythroid 2-related factor-2 signaling pathway to protect B16 cells from oxidative damage. The content of kaempferol-3-O-sophoroside and quercetin-3-O-sophoroside was higher in saffron petals than in other parts of saffron. The kaempferol and quercetin obtained from saffron petals could enhance the activity of antioxidant enzyme NQO1 and T-AOC in B16 cells. This indicated that saffron petals, as a potential functional food, may prevent diseases caused by oxidative stress. © 2024 Society of Chemical Industry.

Transcriptome-wide Characterization of Novel WRKY Family Genes Engaged in Crocin Biosynthesis in Crocus sativus

Crocins comprise a family of hydrophilic carotenoids with pharmacological properties that are produced in significant quantities in stigma of Crocus sativus. Although the biosynthesis pathway of crocins has been sufficiently elucidated, there is a paucity of information regarding how transcription factors (TFs) regulate crocin biosynthesis in various stigma developmental stages. WRKY TFs play a role in modulating carotenoid/apocarotenoid metabolism. To provide an overview of the WRKY family in Crocus sativus (CsWRKY) and characterize candidate CsWRKY TFs involved in the biosynthesis of crocins, CsWRKY genes were identified from RNA-sequenced stigma at different developmental stages. A phylogenetic analysis was performed to characterize their evolutionary interrelatedness. A coexpression analysis of CsWRKY genes and crocin biosynthesis-related genes was performed. A quantitative real-time polymerase chain reaction was used to corroborate the expression level of CsWRKY TFs in various tissue and at different developmental stages. A total of 34 CsWRKY TFs were identified from the stigma of C. sativus. The CsWRKY TFs, together with their orthologs from Arabidopsis, were clustered into group I, II, or III following phylogenetic analysis. A correlation analysis revealed that the expressions of the TFs CsWRKY1, CsWRKY2, CsWRKY8, CsWRKY10, CsWRKY15, and CsWRKY28 were strongly related to the expression of crocin biosynthesis-related genes CsBCH, CsCCD2L, CsALDH, and CsUGT. CsWRKY2, CsWRKY15, and CsWRKY28 exhibited identical motifs and were stratified into group IId. Transcript levels of candidate CsWRKY genes were higher in stigma than in other tissues and were proportional to the crocin content.

English

The current study was conducted in growth chamber with the aims to examine the effect of different soil types and temperature regimes on quantitative and qualitative traits of saffron and to assess the agro-climatic suitability of selected sites of Poonch Division of Azad Jammu & Kashmir for field scale saffron cultivation. Soil from 07 sites was collected and processed for pot filling. A total of 63 plastic pots (21 for each temperature level i.e., 19, 26 and 32 oC), were filled with soils of each site and incubated separately in growth chambers. The pots were arranged under completely randomized design (CRD) replicated thrice. Results indicated that maximum number of flowers (1.35 per plant), flower fresh weight (29.03 mg per plant), stigma length (1.21 cm), stigma fresh weight (8.97 mg per plant) and stigma yield (8.60 mg per pot) was recorded for Khaigala site (loam textured soil) at 26 oC followed by Trarkhel (sandy loam textured soil) and Abbaspur (loam textured soil) sites. On an average, maximum 58.30 days were taken to initiate flowering in soil of Alisojal site (clay loam textured soil) with flowering period of 17.06 days. The maximum number of daughter corms (2.92 per plant), corm diameter (9.61 mm), corm weight (15.76 g per plant), corm yield (47.28 g per pot), picrocrocin (11.32%) and safranal (0.22%) content was again recorded for Khaigala site (at 26 oC). However, maximum crocin content (14.33%) was noted for Alisojal site (1831 m altitude). The Cluster and PC analyses revealed that Khaigala, Trarkhel and Abbaspur sites produced saffron of better quality and agro-climatically suitable for establishment of saffron. Further, the saffron grown on loam and sandy loam textured soils showed better performance compared to that of clay textured soils.

Transcriptome wide analysis of MADS box genes in Crocus sativus and interplay of CstMADS19-CstMADS26 in orchestrating apocarotenoid biosynthesis

Flowers of Crocus sativus L. are immensely important not only for arrangement of floral whorls but more because each floral organ is dominated by a different class of specialized compounds. Dried stigmas of C. sativus flowers form commercial saffron, and are known to accumulate unique apocarotenoids like crocin, picrocrocin and safranal. Inspite of being a high value crop, the molecular mechanism regulating flower development in Crocus remains largely unknown. Moreover, it would be very interesting to explore any co-regulatory mechanism which controls floral architecture and secondary metabolic pathways which exist in specific floral organs. Here we report transcriptome wide identification of MADS box genes in Crocus. A total of 39 full length MADS box genes were identified among which three belonged to type I and 36 to type II class. Phylogeny classified them into 11 sub-clusters. Expression pattern revealed some stigma up-regulated genes among which CstMADS19 encoding an AGAMOUS gene showed high expression. Transient over-expression of CstMADS19 in stigmas of Crocus resulted in increased crocin by enhancing expression of pathway genes. Yeast one hybrid assay demonstrated that CstMADS19 binds to promoters of phytoene synthase and carotenoid cleavage dioxygenase 2 genes. Yeast two hybrid and BiFC assays confirmed interaction of CstMADS19 with CstMADS26 which codes for a SEPALATA gene. Co-overexpression of CstMADS19 and CstMADS26 in Crocus stigmas enhanced crocin content more than was observed when genes were expressed individually. Collectively, these findings indicate that CstMADS19 functions as a positive regulator of stigma based apocarotenoid biosynthesis in Crocus.

Evaluation of Saffron Quality Using Rapid Quantitative Inspection Technology with Near-Infrared Spectroscopy.

A predictive model utilizing near-infrared spectroscopy was developed to estimate the loss on drying, total contents of crocin I and crocin II, and picrocrocin content of saffron. Initially, the LD values were determined using a moisture-ash analyzer, while HPLC was employed for measuring the total contents of crocin I, crocin II, and picrocrocin. The near-infrared spectra of 928 saffron samples were collected and preprocessed using first derivative, standard normal variable transformation, detrended correction, multivariate scattering correction, Savitzky-Golay smoothing, and mean centering methods. Leveraging the partial least squares method, regression models were constructed, with parameters optimized through a selective combination of the above six preprocessing methods. Subsequently, prediction models for loss on drying, total contents of crocin I and crocin II, and picrocrocin content were established, and the prediction accuracy of the models was verified. The correlation coefficients and root mean square error of loss on drying, total contents of crocin I and crocin II, and picrocrocin content demonstrated high accuracy, with R2 values of 0.8627, 0.8851, and 0.8592 and root mean square error values of 0.0260, 0.0682, and 0.0465. This near-infrared prediction model established in the present study offers a precise and efficient means of assessing loss on drying, total contents of crocin I and crocin II, and picrocrocin content in saffron and is useful for the development of a rapid quality evaluation system.

Potential of cold plasma pretreatment for preserving biochemical attributes and ensuring the microbiological safety of saffron stigma.

Saffron, similar to numerous other agricultural commodities, is susceptible to microbial contamination during cultivation and postharvest handling. Cold plasma treatment has emerged as an effective method for microbial inactivation while preserving food quality. The aim of this research was to preserve the color integrity and minimize the presence of microorganisms in dried saffron stigma by implementing cold plasma pre-treatment. Process parameters were optimized using the response surface method (RSM), considering the type of atmosphere (argon and air), plasma exposure time (1, 5, and 10 min), and plasma power (40, 70, and 100 W) as independent variables. The objectives were to maximize crocin content and minimize the total microbial load. The analysis of the response surface revealed that the argon atmosphere had a more significant impact on reducing microbial contamination than air, and an increase in plasma exposure time led to a decrease in microbial load. The maximum reduction in microbial load, by 0.9 logarithmic cycles compared to the control, was achieved with a 10-min treatment at 40 W power. Extended plasma exposure durations led to a minor reduction in the color, taste, aroma, and antioxidant properties of saffron stigma. Specifically, the color, taste, and aroma decreased by 0.5%, 0.5%, and 0.08%, respectively, with longer plasma exposure times. The antioxidant activity decreased by 0.64% with prolonged exposure time. However, the plasma-treated samples did not show any signs of Escherichia coli, mold, or yeast. Furthermore, our findings demonstrated that the type of atmosphere significantly influenced the reduction of infection and maintenance of saffron stigma's color quality. Cold plasma pretreatment holds promise as a viable method for preserving the physicochemical attributes of saffron while effectively reducing microbial contamination.

Saffron of Taliouine and new extension areas in Morocco: farmers surveys and physicochemical analysis

Saffron, the most expensive spice in the world, is a local product of socio-economic importance for developing many rural areas of Morocco, of which Taliouine is the main production area. The views on factors affecting the quality and yield of farmers practicing saffron cultivation are of great importance, and there is currently no research on this topic in the literature. This work aims to evaluate the quality of saffron samples collected at Taliouine (4 samples) and the main extension areas (4 samples) and to survey producers on the characteristics of quality and the main factors involved. The physicochemical analyses were carried out according to the technical specifications of the ISO/TS 3632-2 standard, making it possible to classify the samples according to the Moroccan standard: NM 08.1.037/2007, in categories I or II. Analysis of variance revealed the presence of significant differences (p ≤ 0.05) between E440nm1%\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${\ ext{E}}_{440\ ext{ nm }}^{1{\\%}}$$\\end{document}(specific crocin extinction) ranging from 313.77 to 351.36 of the four samples from the Taliouine area and the E440nm1%\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${\ ext{E}}_{440\ ext{ nm }}^{1{\\%}}$$\\end{document} varying from 204.04 to 492.87 of the four samples from the other regions, between the E257nm1%\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${\ ext{E}}_{257\ ext{ nm}}^{1{\\%}}$$\\end{document} (specific picrocrocin extinction) of the four samples of the Taliouine zone changing from 138.06 to 169.39 and the E257nm1%\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${\ ext{E}}_{257\ ext{ nm}}^{1{\\%}}$$\\end{document} ranging from 134.90 to 219.18 of the four samples from the other zones and finally between the E330nm1%\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${\ ext{E}}_{330\ ext{ nm}}^{1{\\%}}$$\\end{document} (specific safranal extinction) varying from 65.93 to 101.63 of the four samples from the Taliouine zone and the E330nm1%\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${\ ext{E}}_{330\ ext{ nm}}^{1{\\%}}$$\\end{document} of the four samples of the other zones ranging from 89.98 to 119.04. It should be noted that the sample from abroad is fraudulent and non-compliant for almost all the chemical characteristics. Cross-analysis of producer survey data and chemical analysis showed the presence of correlations between stigma length and altitude (r = 0.763*), between crocin content and altitude (r = 0.856*), and between safranal content and humidity (r = 0.722**). According to the producers, the yield remains influenced by the size of the bulbs and the leading cultural practices.Graphical

Discrimination between raw and ginger juice processed Fructus Gardeniae based on UHPLC-Q-TOF-MS and Heracles NEO ultra-fast gas phase electronic nose.

Fructus Gardeniae (ZZ), a traditional Chinese herb, has been used in treating patients with jaundice, inflammation, etc. When mixed with ginger juice and stir-baked, ginger juice-processed Fructus Gardeniae (JZZ) is produced, and the chemical compositions in ZZ would be changed by adding the ginger juice. To illuminate the differential components between ZZ and JZZ. HPLC, UHPLC-Q-TOF-MS, and Heracles NEO ultra-fast gas phase electronic nose were applied to identify the differential components between ZZ and JZZ. HPLC fingerprints of ZZ and JZZ were established, and 24 common peaks were found. The content determination results showed that the contents of shanzhiside, geniposidic acid, genipin-1-β-D-gentiobioside and geniposide increased, while the contents of crocin I and crocin II decreased in JZZ. By UHPLC-Q-TOF-MS, twenty-six possible common components were inferred, among which 11 components were different. In further investigation, eight components were identified as the possible distinctive non-volatile compounds between ZZ and JZZ. By Heracles NEO ultra-fast gas phase electronic nose, four substances were inferred as the possible distinctive volatile compounds in JZZ. Shanzhiside, caffeic acid, genipin-1-β-D-gentiobioside, geniposide, rutin, crocin I, crocin II, and 4-Sinapoyl-5-caffeoylquinic acid were identified as the possible differential non-volatile components between ZZ and JZZ. Aniline, 3-methyl-3-sulfanylbutanol-1-ol, E-3-octen-2-one, and decyl propaonate were inferred as the possible distinctive volatile compounds in JZZ. This experiment explored a simple approach with objective and stable results, which would provide new ideas for studying decoction pieces with similar morphological appearance, especially those with different odors.

Evaluation of Crocin Content and In Vitro Antioxidant and Anti-Glycation Activity of Different Saffron Extracts.

Crocin, a glycoside carotenoid that exhibits several health benefits, is mainly obtained from saffron (Crocus sativus L.), whose quality and content of phytochemicals can be strongly affected by environmental conditions. Therefore, in this work, the crocin content and in vitro antioxidant activity of saffron extracts obtained from three different varieties (Greek, Sicilian, and Iranian saffron) were assessed. Crocin content in saffron extracts was quantified via ultra-performance liquid chromatography coupled with mass spectrometry. The antioxidant activity of saffron extracts was evaluated using the oxygen radical absorbance capacity (ORAC) assay and nitric oxide (NO) radical scavenging test. The Maillard reaction was used to assess anti-glycation activity. Although the Sicilian and Iranian saffron extracts contained higher amounts of crocin (128 ± 6 ng/mL and 126 ± 4 ng/mL, respectively) compared to the Greek extracts (111 ± 2 ng/mL), ORAC values (50.9 ± 0.5) and % NO inhibition (35.2 ± 0.2) were higher for the Greek variety, which displayed a total phenolic content about two-fold greater than that of the other two extracts. Sicilian and Greek saffron had similar anti-glycation activities, while Iranian saffron was less effective. These results suggest that the antioxidant activity of saffron extracts could be ascribed to their naturally occurring complex mixture of phytochemicals, deserving further investigation as supplements to prevent pathological conditions induced by radical species.

Simultaneous determination of crocin and crocetin in food samples by highperformance liquid chromatography (HPLC-PDA)

Crocin and crocetin are two bioactive compounds found in saffron and gardenia. In the food industry, due to their natural yellow or orange color, they are used as food colouring. The study was carried out to optimize the high-performance liquid chromatography combined with a photodiode detector (HPLC-PDA) to simultaneously determine the crocin and crocetin content in food. Analytical samples were ultrasonicated with ACN:H2O (1:1, v/v) solvent for 30 minutes. The sample solution was determined by HPLC – PDA with the following conditions: Sunfire C18 column (250 mm x 4.6 mm; 5 µm) mobile phase consisting of 0.1% formic acid and ACN using gradient program. The method has good specificity, the standard curves of crocin and crocetin have correlation coefficients R2 > 0.999, respectively, repeatability and recovery meet AOAC requirements. The limit of detection (LOD) for  crocin and crocetin were 0.5 mg/kg and 0.25 mg/kg, respectively. The method has been applied to simultaneously determine crocin and crocetin in 15 real samples and results in 13 out of 15 samples detecting crocin content, 11 out of 15 samples detecting crocetin content, and in 2 out out of 15 samples. neither crocin nor crocetin was detected. The gardenia powder group contained the highest content of crocin and crocetin (23.5 g/100g – 24.4 g/100g) and dried lychee samples - sun-dried or dried gardenia contains the second highest concentrations of crocin and crocetin in the food group.

Arbuscular mycorrhizal symbiosis modulates the apocarotenoid biosynthetic pathway in saffron

Crocus sativus L. (saffron) has been propagated for millennia to produce the precious spice saffron from the red stigmas. The inebriant organoleptic and bioactive properties mainly depend on the content of crocins (dyeing capacity), picrocrocin (flavor), and safranal (aroma), apocarotenoids deriving from zeaxanthin. In this study, an integrated biochemical and molecular analysis was carried out on fresh saffron stigmas to investigate the influence exerted by the arbuscular mycorrhizal fungus (AMF) Rhizophagus intraradices on the production of the main saffron apocarotenoids responsible for the properties of the spice. Since mineral enrichment due to AM symbiosis has been related to changes in the secondary metabolism of plants, the mineral content of saffron corms at flowering was also analyzed. Rare arbuscules (AMF trade structures) were found in mycorrhized plants. However, the expression of D27, CCD7, and NCED involved in the synthesis of strigolactones (SLs) and abscisic acid (ABA), which promote AM symbiosis, did not change in the stigmas. The transcription of β-LYC and CCD4a/b was not affected by AMF, whereas that of CCD2, which encodes the key enzyme producing major apocarotenoids, was upregulated. The crocin content was reduced in treated plants even if the expression of ALDH, UGT74AD1, and UGT91P3, involved in crocin synthesis, did not change. Conversely, UGT709G1, implicated in picrocrocin synthesis, was overexpressed in the inoculated plants, thus the safranal content was increased in the spice.

Natural variations in Crocus sativus lycopene epsilon cyclase (CstLcyE) alter carotenoid/apocarotenoid content and stress tolerance

Crocus sativus is an economically and pharmaceutically important crop. The dried stigmas of C. sativus flowers constitute saffron, the only commercial source of apocarotenoids like crocin, picrocrocin, and safranal. Some elite clones of C. sativus with higher crocin content were identified from plant populations over three years. Sequencing approach demonstrated that mutations are present in CstLycE. The C. sativus apocarotenoids are formed by cleavage of β-branch carotenoids. Cyclization of lycopene is the key branch point in carotenoid/apocarotenoid pathway. While lycopene β-cyclase converts lycopene into β-carotene, combined action of β and ε-cyclases forms α-carotene. Towards this, the mutant populations with critical mutations in CstLycE result in loss of function, and hence the metabolic flux was diverted towards β-branch. In-silico analysis and site-directed mutagenesis confirmed the critical nature of these mutations. To ascertain the role of CstLycE, transient over-expression and RNAi lines of CstLcyE in C. sativus were generated, and we observed enhanced β-branch carotenoid and apocarotenoid content in RNAi lines. The effect of CstLcyB and CstLcyE was compared, and CstLcyE RNAi lines had a more profound impact on apocarotenoid accumulation than CstLcyB over-expression lines. CstLcyE-RNAi plants also accumulated more abscisic acid and were more tolerant to abiotic stresses.

Transcriptome and metabolome analysis revealed the changes of Geniposide and Crocin content in Gardenia jasminoides fruit.

Gardenia jasminoides Ellis is a perennial evergreen shrub of G. jasminoides of Rubiaceae. Geniposide and Crocin are important components in the fruit of G. jasminoides. In addition to being used as medicinal materials, they are also widely used in food, medicine, cosmetics, and other fields. They have high medicinal value, economic value, and ornamental value. However, at present, the utilization rate of G. jasminoides resources is low, mainly focused on germplasm cultivation, primary processing, and clinical pharmacology, and there are few studies on the quality of Gardenia fruit. Based on transcriptome sequencing and metabolic group analysis, the morphological and structural changes of Gardenia fruit with young fruit, middle fruit, and ripe fruit were analyzed, and the formation mechanism and content changes of Geniposide and Crocin in Gardenia fruit were studied. The content of Geniposide decreased with the development of fruit, so did the expression of the main structural gene GES, G10H, and IS in its synthesis pathway, while the content of Crocin increased with the development of fruit, and the expression of the main structural gene CCD, ALDH, and UGT in its synthesis pathway also increased. The relationship between the morphological structure of G. jasminoides and the accumulation of Geniposide and Crocin was summarized. This study not only provides a theoretical basis for the mining and utilization of Geniposide and Crocin, but also provides a theoretical basis for genetic background for the identification and cloning of bioactive substances in gardenia fruit in future. At the same time, it provides support for increasing the dual-use value of G. jasminoides and breeding excellent germplasmresources.

Fabrication of protein crocin nanocomplex and its addition in proso millet flour for development of functional snacks with anti‐diabetic, anti‐hypertensive and antioxidant properties

SummaryThe current study was undertaken to develop novel gluten‐free functional snacks using proso millet flour with the addition of nano‐encapsulated crocin in sorghum (SPCS), pearl millet (PPCS) and foxtail millet (FPCS) protein nanoparticles. The crocin content was well retained (58.08%–77.67%) in the developed snacks. Characteristic properties like nutraceutical potential, rheological, sensory and physical attributes were evaluated. The extruded snacks showed good antioxidant (DNA damage inhibition assay), anti‐diabetic (alpha amylase inhibition assay) and anti‐hypertensive (angiotensin converting enzyme assay) properties. The developed snacks (P > 0.05) showed higher elastic behaviour (G′) compared to viscous behaviour (G″). Hardness value for functional snacks were found to be in the range of 407–115 N. The control snacks showed significantly higher expansion ratio (348.76%) compared to functional snacks. The developed novel functional snacks appeared darker than control snacks as evaluated by colorimetric assay. The overall acceptability of the snacks was found to be equal on a 9‐point hedonic scale.

Elucidation of Geniposide and Crocin Accumulation and Their Biosysnthsis-Related Key Enzymes during Gardenia jasminoides Fruit Growth.

Gardenia jasminoides fruits are extensively grown worldwide, with a large harvest, and its major medicinal ingredients are geniposide and crocins. Research on their accumulation and biosynthsis-related enzymes is rare. In this study, the accumulation of geniposide and crocin of G. jasminoides fruits at different developmental stages were clarified by HPLC. The highest cumulative amount of geniposide was 2.035% during the unripe-fruit period, and the highest content of crocin was 1.098% during the mature-fruit period. Furthermore, transcriptome sequencing was performed. A total of 50 unigenes encoding 4 key enzymes related in geniposide biosynthsis pathways were screened, and 41 unigenes encoding 7 key enzymes in the pathways of crocin were elucidated. It was found that the expression levels of differentially expressed genes of DN67890_c0_g1_i2-encoding GGPS, which is highly related to geniposide biosynthesis, and DN81253_c0_g1_i1-encoding lcyB, DN79477_c0_g1_i2-encoding lcyE, and DN84975_c1_g7_i11-encoding CCD, which are highly related to crocin biosynthesis, were consistent with the accumulation of geniposide and crocin content, respectively. The qRT-PCR results showed that the trends of relative expression were consistent with transcribed genes. This study provides insights for understanding the geniposide and crocin accumulation and biosynthsis during fruit development in G. jasminoides.

Diversity of Culturable Endophytic Fungi in Crocus sativus and Their Correlation with Crocin Content.

The presence and diversity of endophytic fungi associated with host plants are important not just for host plant growth and defense, but also impact the production of medicinal secondary metabolites. However, the correlation between endophytic fungi and crocin production in Crocus sativus (CS) remains underexplored. Here, we explore the relationship between endophytic fungal diversity and crocin content among different CS tissues and field sites. Specifically, we isolated endophytic fungi from five different field sites (Shanghai, Jiande, Huzhou, Anhui, and Hebei) and five different tissues (corm, scape, leaf, petal, and stigma) and analyzed fungal community diversity, richness, and evenness. We identified a total of 32 endophytic fungal taxa, assigned to 7 orders within 4 classes (Eurotiomycetes, Agaricomycetes, Dothideomycetes, and Sordariomycetes). The most dominant order was Eurotiales, and the most dominant genera were Penicillium and Talaromyces. Species richness tended to be highest in belowgrown tissues, such as corm and scape. Additionally, several fungal taxa were found to be either site- or tissue-specific. Three genera in particular were correlated with crocin content: Penicillium, Sistotrema, and Bjerkandera. Given the fact that endophytic microorganisms can both promote the production of secondary metabolites in host plants and potentially produce secondary metabolites themselves, further study is required to understand the mechanistic relationship between these and other fungal genera and crocin production.

Effect of different extraction methods on saffron antioxidant activity, total phenolic and crocin contents and the protective effect of saffron extract on the oxidative stability of common vegetable oils

Saffron consists of bioactive compounds with health-promoting properties and is mainly used in medicine, flavoring and coloring. In this study, we aimed to investigate the effect of extraction methods on the antioxidant activity of saffron (Crocus sativus L.) extracts (SE) and to evaluate the antioxidant performance of SE in vegetable oils. Saffron stigmas were extracted in water, ethanol, methanol, and their combinations using maceration extraction (ME), ultrasonic-assisted extraction (UAE), microwave-assisted extraction (MAE), and the combination of UAE with MAE. The results showed that the sample extracted by methanol/water (50:50) using the combination of UAE with MAE methods had the highest amount of total phenolic content (31.56 mg/g GAE) and antioxidant activity (83.24% inhibition). The extract with the highest antioxidant activity was freeze-dried before incorporation into oil samples. Freeze-dried SE contained trans-crocin-4 and trans-crocin-3 (most abundant constituents), kaempferol, and picrocrocin. Moreover, the addition of SE at 1000 ppm resulted in a significant increase in the oxidative stability of canola (CAO), sunflower (SO), and corn oil (COO).

Extraction Optimization of Crocin from Gardenia (Gardenia jasminoides Ellis)Fruits Using Response Surface Methodology and Quality Evaluation of Foam-Mat Dried Powder

The crocin in gardenia, as a medical plant, has drawnthe attention of researchers and scientists due to its color and high antioxidant activity. To optimize the extraction parameters of crocin from gardenia fruits, response surface methodology (RSM) was employed.The effects of four independent variables, namely extraction temperature (45–55 °C), time (40–60 min), percentage of gardenia fruits(15–25%), and ethanol concentration (50–60%),on a crocin compound were investigated. The extract from the gardenia fruit was dried at different temperatures (55–70 °C) by the foam-mat drying method. The optimal extraction parameters were an extraction temperature of 55 °C, time of 57 min, percent of fruits in solvent at 24%, and an ethanol concentration of 56%. The results showed that the dried gardenia powder had maintained the crocin content well(6.64 mg/g), and the product with low water activity and moisture content of 0.33 and 5.72%, respectively, is suitable for storage. The foam-mat dried product also maintains the natural color and characteristics inherent in the raw materials, which could also be used as supplemental ingredients for other food industries.

Saffron benefits for eye diseases

AbstractGlaucoma is a neurodegenerative disease characterized by the loss of retinal ganglion cells (RGCs). An increase in the intraocular pressure is the principal risk factor but controlling this pressure does not always prevent glaucomatous damage, so other harmful factors would be implicated in this pathologic process. Several mechanisms have been proposed that leads to the RGC death in glaucoma, including glutamate excitotoxicity, aggregation of misfolded proteins, mitochondrial dysfunction, oxidative stress, neurotrophic deprivation, and neuroinflammation, among others. Therefore, early diagnosis of neurodegeneration, understanding of its pathogenic mechanisms, and the development of new neuroprotective therapies represent major challenges for treating this disease.During the inflammatory process that occurs in glaucoma, retinal glial cells, both microglia and macroglia (Müller cells and astrocytes), become activated and release factors that can be neuroprotective in some cases and neurodegenerative in others. Cytokine‐ and chemokine‐mediated signalling engages in the neuroinflammatory process that leads to retinal ganglion cell (RGC) damage in glaucoma. Thus, a substance with anti‐inflammatory activity may protect against RGC degeneration.Increasing evidence from both experimental models and clinical studies in patients supports the neuroprotective effect of saffron components in neurodegenerative conditions and in retinal neurodegenerative pathologies, due to its important anti‐inflammatory, anti‐apoptotic, and antioxidant properties.This study investigated the neuroprotective and anti‐inflammatory effects of a hydrophilic saffron extract standardized to 3% crocin content in a mouse model of unilateral, laser‐induced ocular hypertension (OHT). For this purpose, morphological signs of microglial activation and the expression of P2RY12, which demonstrates the increased microglial activation, were analysed. In addition, the possible regulation of cytokine and myokine production was also studied. Treatment with saffron extract decreased microglial signs of activation including reduction of microglial cell numbers and morphological features of its activation, including decrease soma size and process retraction, both in OHT and in contralateral eyes. In addition, also partially reversed OHT‐induced down‐regulation of P2RY12 and prevented retinal ganglion cell death in OHT eyes. Furthermore, most of the concentration of proinflammatory cytokines (IL‐1β, IFN‐γ, TNF‐α, and IL‐17), anti‐inflammatory cytokines (IL‐4 and IL‐10), Brain‐derived Neurotrophic Factor (BDNF), Vascular Endothelial Growth Factor (VEGF), and fractalkine were unaffected in response to laser‐induced OHT in both the OHT eye and its contralateral eye in animals treated with saffron extracts. Only IL‐6 levels were significantly increased in the OHT eye one day after laser induction compared with the control group.These results differed from those observed in this OHT glaucoma model in animals untreated with saffron, where microglial activation and down‐regulation of P2RY12 was more intense, and the significant changes in cytokine levels in both eyes in response to IOP increase were observed.All these findings indicate that oral administration of saffron extract could be beneficial in glaucoma by decreasing the neuroinflammation associated with increased intraocular pressure, reducing microglial activation, regulating the production of proinflammatory cytokines, VEGF, and fractalkine and preventing retinal ganglion cell death.

Quantification of apocarotenoids in commercial Indian (Kashmiri) saffron using UV-Vis spectroscopy and HPLC analysis

Saffron is considered as the most expensive spice in the world. Due to low production, high demand and high cost, saffron is very prone to adulteration for economic benefits while putting public health at risk. The most significant characteristic for determining the quality of the saffron is coloring strength (crocin content), which is determined by measuring UV-Vis absorption at 440 nm in the aqueous preparations of this spice. Picrocrocin and safranal are other key components used to determine saffron quality. This article aims to examine the quality of commercial saffron obtained from various geographical locations of Kashmir (India) by determining their apocarotenoid content using UV-Vis spectrophotometry followed by high-performance liquid chromatography (HPLC) to determine the concentration of saffron metabolites (crocin, picrocrocin and safranal). A total of 31 samples from different origins were used in this study. The UV-Vis spectrophotometric results showed that among 31, only 14 samples fell into grade I, while 9 samples fell in grade II and 5 samples fell in grade III of the ISO category. The remaining 3 samples could not satisfy ISO standards, which indicates that these samples were adulterated. The determination of apocarotenoid content using HPLC analysis varied significantly among samples. These variations may be due to different drying and storage conditions or adulteration.