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Molecular cloning and expression of Cro s 1: an occupational allergen from saffron pollen (Crocus sativus).

ABSTRACT: BACKGROUND:The cultivation of saffron is expanding through the southeast of Iran, and allergy to saffron pollen occurs in workers involved in processing this plant. We aimed to clone, sequence and express a major allergen involved in saffron pollen allergy, and to compare the recombinant with the natural allergen. METHODS:The N-terminal amino acid sequence of Cro s 1, an allergen from saffron pollen, was determined after immunoblotting. The cDNA encoding for this allergen was cloned by PCR utilizing a primer based on the N-terminal amino acid sequence. Recombinant Cro s 1 (rCro s 1) was expressed as a soluble protein in Pichia pastoris and purified to homogeneity by gel filtration. Inhibition of IgE binding to rCro s 1 by pollen extract was analyzed by ELISA. SECTION TITLE:The allergen Cro s 1 was identified from saffron pollen extracts and cloned by PCR. Cro s 1 cDNA defined an acidic polypeptide with homology to pollen proteins from Chenopodium album and Ligastrum vulgaris. The rCro s 1 was expressed in P. pastoris at 28 mg/l. Saffron pollen extract inhibited the binding of patient serum IgE to rCro s 1. CONCLUSION:We identified and cloned the first Crocus sativus pollen allergen. rCro s 1 cDNA shows a very high homology with Che a 1, the major allergen of lamb's-quarter, Chenopodium album, Caryophyllales, pollen (97%). Cro s 1 is a useful tool for specific diagnosis and structural studies of occupational allergy to saffron.

SUBMITTER: Varasteh AR

PROVIDER: S-EPMC4757073 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature

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Epigenetic variability among saffron crocus (Crocus sativus L.) accessions characterised by different phenotypes

Project description:This work represents the first epigenomic study carried out on saffron crocus. Five accessions of saffron, showing differences in tepal pigmentation, yield of saffron and flowering time, were analysed at the epigenetic level by applying a methylation-sensitive restriction enzyme-sequencing (MRE-seq) approach. Five accession-specific hypomethylomes plus a reference hypomethylome, generated by combining the sequence data from the single accessions, were obtained. Assembled sequences were annotated against existing online databases. In the absence of the Crocus genome, the rice genome was mainly used as the reference as it is the best annotated genome among monocot plants. Comparison of the hypomethylomes revealed many differentially methylated regions, confirming the high epigenetic variability present among saffron accessions, including sequences encoding for proteins that could be good candidates to explain the accessions’ alternative phenotypes. In particular, transcription factors involved in flowering process (MADS-box and TFL) and for the production of pigments (MYB) were detected. Finally, by comparing the generated sequences of the different accessions, a high number of SNPs, likely having arisen as a consequence of the prolonged vegetative propagation, were detected, demonstrating surprisingly high genetic variability. Gene ontology (GO) was performed to map and visualise sequence polymorphisms located within the GOs and to compare their distributions among different accessions. As well as suggesting the possible existence of alternative phenotypes with a genetic basis, a clear difference in polymorphic GO is present among accessions based on their geographic origin, supporting a possible signature of selection in the Indian accession with respect to the Spanish ones.

2021-03-24 | GSE166350 | GEO

AFLP and MS-AFLP analysis of the variation within saffron crocus (Crocus sativus L.) germplasm.

Project description:The presence and extent of genetic variation in saffron crocus are still debated, as testified by several contradictory articles providing contrasting results about the monomorphism or less of the species. Remarkably, phenotypic variations have been frequently observed in the field, such variations are usually unstable and can change from one growing season to another. Considering that gene expression can be influenced both by genetic and epigenetic changes, epigenetics could be a plausible cause of the alternative phenotypes. In order to obtain new insights into this issue, we carried out a molecular marker analysis of 112 accessions from the World Saffron and Crocus Collection. The accessions were grown for at least three years in the same open field conditions. The same samples were analysed using Amplified Fragment Length Polymorphism (AFLP) and Methyl Sensitive AFLP in order to search for variation at the genetic (DNA sequence) and epigenetic (cytosine methylation) level. While the genetic variability was low (4.23% polymorphic peaks and twelve (12) effective different genotypes), the methyl sensitive analysis showed the presence of high epigenetic variability (33.57% polymorphic peaks and twenty eight (28) different effective epigenotypes). The pattern obtained by Factorial Correspondence Analysis of AFLP and, in particular, of MS-AFLP data was consistent with the geographical provenance of the accessions. Very interestingly, by focusing on Spanish accessions, it was observed that the distribution of the accessions in the Factorial Correspondence Analysis is not random but tends to reflect the geographical origin. Two clearly defined clusters grouping accessions from the West (Toledo and Ciudad Real) and accessions from the East (Cuenca and Teruel) were clearly recognised.

| S-EPMC4401542 | biostudies-literature

Transcriptome profiling of the flowering transition in saffron (Crocus sativus L.).

Project description:Saffron, derived from the stigma of Crocus sativus, is not only a valuable traditional Chinese medicine but also the expensive spice and dye. Its yield and quality are seriously influenced by its flowering transition. However, the molecular regulatory mechanism of the flowering transition in C. sativus is still unknown. In this study, we performed morphological, physiological and transcriptomic analyses using apical bud samples from C. sativus during the floral transition process. Morphological results indicated that the flowering transition process could be divided into three stages: an undifferentiated period, the early flower bud differentiation period, and the late flower bud differentiation period. Sugar, gibberellin (GA3), auxin (IAA) and zeatin (ZT) levels were steadily upregulated, while starch and abscisic acid (ABA) levels were gradually downregulated. Transcriptomic analysis showed that a total of 60 203 unigenes were identified, among which 19 490 were significantly differentially expressed. Of these, 165 unigenes were involved in flowering and were significantly enriched in the sugar metabolism, hormone signal transduction, cell cycle regulatory, photoperiod and autonomous pathways. Based on the above analysis, a hypothetical model for the regulatory networks of the saffron flowering transition was proposed. This study lays a theoretical basis for the genetic regulation of flowering in C. sativus.

| S-EPMC7295807 | biostudies-literature

Bacillus subtilis FZB24 affects flower quantity and quality of saffron (Crocus sativus).

Project description:The effect of Bacillus subtilis FZB24 on saffron ( Crocus sativus L.) was studied using saffron corms from Spain and the powdered form of B. SUBTILIS FZB24(R). Corms were soaked in water or in B. subtilis FZB24 spore solution for 15 min before sowing. Some corms were further soil drenched with the spore solution 6, 10 or 14 weeks after sowing. Growth and saffron stigma chemical composition were measured. Compared to untreated controls, application of B. subtilis FZB24 significantly increased leaf length, flowers per corm, weight of the first flower stigma, total stigma biomass; microbe addition also significantly decreased the time required for corms to sprout and the number of shoot sprouts. Compared to the controls, picrocrocin, crocetin and safranal compounds were significantly increased when the plants were soil drenched with the spore solution 14 weeks after sowing; in contrast crocin was highest in untreated controls. Results of this study suggest that application of B. subtilis FZB24 may provide some benefit to saffron growers by speeding corm growth (earlier shoot emergence) and increasing stigma biomass yield by 12 %. While some treatment conditions also increased saffron chemical composition, these were generally not the same treatments that simultaneously improved growth yields and thus, more study is required.

| S-EPMC3947403 | biostudies-literature

Smooth muscle relaxant activity of Crocus sativus (saffron) and its constituents: possible mechanisms.

Project description:Saffron, Crocus sativus L. (C. sativus) is rich in carotenoids and used in traditional medicine for treatment of various conditions such as coughs, stomach disorders, amenorrhea, asthma and cardiovascular disorders. These therapeutic effects of the plant are suggested to be due to its relaxant effect on smooth muscles. The effect of C. sativus and its constituents on different smooth muscles and the underlying mechanisms have been studied. Several studies have shown the relaxant effects of C. sativus and its constituents including safranal, crocin, crocetin and kaempferol on blood vessels. In addition, it was reported that saffron stigma lowers systolic blood pressure. The present review highlights the relaxant effects of C. sativus and its constituents on various smooth muscles. The possible mechanisms of this relaxing effect including activation of ß2-adrenoceptors, inhibition of histamine H1 and muscarinic receptors and calcium channels and modulation of nitric oxide (NO) are also reviewed.

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Crocus sativus isolate:Saffron JU | cultivar:Saffron

Project description:Crocus sativus isolate:Saffron JU | cultivar:Saffron Genome sequencing and assembly

| PRJNA734464 | ENA

Sustainable Processing of Floral Bio-Residues of Saffron (Crocus sativus L.) for Valuable Biorefinery Products.

Project description:Tepals constitute the most abundant bio-residues of saffron (Crocus sativus L.). As they are a natural source of polyphenols with antioxidant properties, they could be processed to generate valuable biorefinery products for applications in the pharmaceutical, cosmetic, and food industries, becoming a new source of income while reducing bio-waste. Proper storage of by-products is important in biorefining and dehydration is widely used in the herb sector, especially for highly perishable harvested flowers. This study aimed to deepen the phytochemical composition of dried saffron tepals and to investigate whether this was influenced by the extraction technique. In particular, the conventional maceration was compared with the Ultrasound Assisted Extraction (UAE), using different solvents (water and three methanol concentrations, i.e., 20%, 50%, and 80%). Compared to the spice, the dried saffron tepals showed a lower content of total phenolics (average value 1127.94 ± 32.34 mg GAE 100 g-1 DW) and anthocyanins (up to 413.30 ± 137.16 mg G3G 100 g-1 DW), but a higher antioxidant activity, which was measured through the FRAP, ABTS, and DPPH assays. The HPLC-DAD analysis detected some phenolic compounds (i.e., ferulic acid, isoquercitrin, and quercitrin) not previously found in fresh saffron tepals. Vitamin C, already discovered in the spice, was interestingly detected also in dried tepals. Regarding the extraction technique, in most cases, UAE with safer solvents (i.e., water or low percentage of methanol) showed results of phenolic compounds and vitamin C similar to maceration, allowing an improvement in extractions by halving the time. Thus, this study demonstrated that saffron tepals can be dried maintaining their quality and that green extractions can be adopted to obtain high yields of valuable antioxidant phytochemicals, meeting the requirement for a sustainable biorefining.

| S-EPMC8001949 | biostudies-literature

Molecular species fingerprinting and quantitative analysis of saffron (<i>Crocus sativus</i> L.) for quality control by MALDI mass spectrometry.

Project description:Herein we describe a rapid, simple, and reliable method for the quantitative analysis and molecular species fingerprinting of saffron (Crocus sativus L.) by direct MS and MS/MS analysis. Experimentally, powdered saffron was subjected to a brief treatment with a 0.3% TFA water/acetonitrile solution, and the resulting mixture was directly placed on the MALDI plate for analysis. This approach allowed the detection of the commonly observed crocins C-1-C-6 and flavonols, together with the identification of the unknown highly glycosylated crocins C-7, C-8 and C-9, and carotenoid-derived metabolites. The strategy endorsed the simultaneous detection and characterization of saffron and adulterant markers using crude extracts of the adulterant itself and synthetic sets of adulterated authentic saffron samples. The implementation of the strategy was to measure the amount of an unknown adulterant from the crude extract using curcumin as a non-isotopic isobaric internal standard. The relationship between the saffron and curcumin molar ratios were established with a correlation coefficient of 0.9942. The ANOVA regression model was significant, F(1, 72) = 13 595.82, p < 0.001, y = (0.0116 ± 0.0001)x + (-0.1214 ± 0.0086). No matrix effects were observed and good results were obtained with respect to instrumental repeatability (*RSD% < 2%) and LOD (1.1%). The analysis of commercial samples of saffron using the proposed approach showed the suitability of the method for routine analysis (minimal sample preparation and very short measuring time per sample).

| S-EPMC9088749 | biostudies-literature

Deciphering genetic diversity analysis of saffron (<i>Crocus sativus</i> L.) using RAPD and ISSR markers.

Project description:The existence of genetic diversity in Crocus sativus has globally remained a mystery till date. The study investigated PCR based DNA amplification profile of saffron using ISSR and RAPD based primers. A total of 38 amplicons were generated by ISSR primers in the range from 7 to 12 with an average of 9.50 bands per primer. 20 bands were found to be polymorphic and 18 were monomorphic with an average percentage of polymorphism as 52.48%. RAPD based amplification revealed a total 161 amplicons, 107 as polymorphic and 54 as monomorphic with an average percentage of polymorphism as 66.44%. Cumulative results of RAPD and ISSR demonstrated that Nei-Li's similarity index ranged between 0.70 and 0.97. The results of AMOVA has revealed 9% of variance among populations and 91% of variance within populations, Φ PT was found as 0.089, which indicates existence of genetic differences though limited. In conclusion, the results indicate that saffron accessions are minimally genetically differentiated, which could be capitalized in future breeding programmes to ameliorate this precious crop.

| S-EPMC7878693 | biostudies-literature

Saffron (Crocus sativus) versus duloxetine for treatment of patients with fibromyalgia: A randomized double-blind clinical trial.

Project description:Objective:Saffron was found efficient and safe in treatment of neuropsychiatric disorders, in particular depression. We compared the efficacy of saffron with duloxetine in treatment of patients with fibromyalgia. Materials and Methods:In this double-blind parallel-group clinical trial, outpatients with fibromyalgia were randomized to receive either saffron 15 mg or duloxetine 30 mg starting with 1 capsule per day in the first week followed by 2 capsules per day from week 2 until the end of week 8. Participants were men and women aged 18-60 years diagnosed with fibromyalgia based on the American College of Rheumatology 2010 criteria who also had a pain score?40 based on visual analogue scale. Participants were excluded in case they had rheumatologic diseases, inflammatory/infectious/autoimmune arthritis, comorbid neuropsychiatric disorders except depressive disorders, pain due to traumatic injuries, drug history of duloxetine or saffron use, current use of psychoactive medications, recent use of muscle relaxants, steroids, opioid analgesics, benzodiazepines, anti-epileptics, or injective analgesics. Primary outcomes included differences in mean score changes from baseline to endpoint between the treatment arms for Hamilton Rating Scale for Depression, Fibromyalgia Impact Questionnaire, and Brief Pain Inventory. Results:Socio-demographic characteristics and baseline scores were similarly distributed between the two treatment arms (2n=46). No significant difference was detected for any of the scales neither in terms of score changes from baseline to endpoint between the two treatment arms (Mean score changes: -4.26 to 2.37; p-values: 0.182-0.900) nor in terms of timetreatment interactions (p-values: 0.209-0.964). Conclusions:Saffron and duloxetine demonstrated comparable efficacy in treatment of fibromyalgia symptoms.

| S-EPMC6235666 | biostudies-other

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