Phylogenetic relationship and sequence diversity of candidate genes involved in anthocyanin biosynthesis pathway in Carthamus species with contrasting seed coat colors.

The morphological structure of seed such as coat color can be considered as effective parameters in the evaluation of resistance to pests. The present study is aimed at achieving these goals: first, to determine the phylogenetic relationship of different species of safflower with different seed coat colors based on three candidate genes in the anthocyanin biosynthesis pathway that encode the early steps (PAL: phenylalanine ammonia-lyase and CHS: chalcone synthase) and the final step (UFGT: flavonoid-3-O-glucosyltransferase); second, based on our previous study on the absence of cyanidin-3-O-glucoside (Cyd-3-glu) in white/brown-seeded genotypes, it can be determined whether the lack of production is related to the absence of genes or the lack of expression. In general, the detection of Cyd-3-glu upstream compounds in all studied safflower genotypes, regardless of the color of the seed coat, can be interpreted as the expression of genes responsible for the synthesis of these compounds in the anthocyanin synthesis pathway. In addition, these findings indicated that the accumulation pattern of the mentioned secondary metabolites could be varied in safflower genotypes according to the seed coat color pattern. Regarding the UFGT gene, the evidence showed that this gene is expressed in safflower genotypes with two different seed coat color patterns, but in each genotype the tendency to produce secondary metabolites is different. Consequently, it seems that UFGT may not only regulate Cyd-3-glu biosynthesis but also involved in biosynthesis of flavonol glucoside in black safflower. Additionally, UFGT only affected flavonol glycosides biosynthesis and had no effect on Cyd-3-glu biosynthesis in white- seeded safflower genotypes.

Determination of bisphenol a migration from food packaging by dispersive liquid-liquid microextraction.

In the current work, a rapid and simple dispersive liquid-liquid microextraction method (DLLME) was used to determine Bisphenol A (BPA). High performance liquid chromatography with the photodiode-array detector (HPLC-DAD) coupled DLLME method was employed to analyze BPA in food samples packaged including cans, paper boxes, and glass jars. The calibration curve was obtained to be in the linear range 0.009-25 ngg-1 with a correlation coefficient of R2 = 0.9981. The mean relative standard deviations (RSDs) was of 5.2% (n = 3). The limit of detection (LOD) and the limit of quantification (LOQ) of the method were obtained to be 0.001 ngg-1 and 0.08 ng.g-1, respectively. In sum, this method presents:•A rapid, simple and efficient modified DLLME method was used to measure BPA in packaged foods.•The advantages of this method were low detection limit, fast preparation, and high BPA recovery.•The DLLME-HPLC method consists of low detection limit and high recoveries to determine BPA in samples.•The results indicated that DLLME -HPLC-DAD was an applied method to measure BPA in food samples.

Seed Polyphenolic Profile, Antioxidative Activity, and Fatty Acids Composition of Wild and Cultivated Carthamus Species.

Total flavonoid content (TFC) and cyanidin-3-glucoside (Cyd-3-glu) of seed and seed coat extract of 16 genotypes from five species of Carthamus were studied, and their major polyphenolic compounds and antioxidant activity of the seed coat extracts were determined using HPLC analysis and DPPH assay, respectively. Additionally, fatty acids composition of the seed oil was analyzed by GC. In general, TFC and Cyd-3-glu content of seed coat extracts were higher than those of seed extracts. A novel breeding line with black seed coat (named A82) depicted lower TFC (3.79 mg QUE/g DW) but higher Cyd-3-glu (24.64 mg/g DW) compared to the white and other seed-pigmented genotypes. DPPH radical scavenging activity showed a strong association with Cyd-3-glu content (r = 0.84), but no correlation with TFC (r = -0.32). HPLC analysis of seed coat extracts revealed that four compounds were dominant constituents including rutin (7.23 - 117.95 mg/100 g DW), apigenin (4.37 - 64.88 mg/100 g DW), quercetin (3.09 - 14.10 mg/100 g DW), and ferulic acid (4.49 - 30.41 mg/100 g DW). Interestingly, the genotype A82 with an appropriate polyunsaturated/saturated fatty acids index (5.46%) and a moderate linoleic fatty acid content (64.70%) had higher nutritional and pharmaceutical value than all the other genotypes.

Safety Assessment of a New Pigmented Safflower Seed Coat (A82) by a Feeding Study on Rat

ABSTRACT Safflower (Carthamus tinctorius L.) is an annual herbaceous plant, cultivated mainly for the seed which is used for edible oil extraction and bird feeding. This study was designed to evaluate the safety of a new pigmented variety of safflower (A82) seeds. The results showed that oral administration of A82 seeds significantly increased the body weight of male rats in a dose-dependent manner (p<0.05). Biochemical tests showed that A82 seeds significantly increased the serum levels of AST (Aspartate aminotransferase) (p<0.05), slightly reduced the serum levels of ALT (Alanine aminotransferase) and significantly reduced ALP (p<0.05) levels in a dose dependent manner. BUN (Blood Urea Nitrogen) and Cr (Creatinine) were not significantly changed in A82 seed treated groups. Also, testosterone levels were not significantly changed by administration of different doses of A82. However, Johnson scoring showed slightly decrease in experimental groups. No organ weight or histological changes were observed in liver, kidney, spleen, heart and brain of A82 seed treated animals. These results indicate that A82 seeds have not any toxic effects in Wistar rats. Future studies are required to clarify the exact mechanism by which A82 seeds alter AST levels and body weight in rat.

Assessment of performance ability of three diagnostic methods for detection of Potato Leafroll virus (PLRV) using different visualizing systems.

To diminish the time required for some diagnostic assays including reverse transcription PCR (RT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP; due to mainly RNA extraction step) and also DAS-ELISA into a minimum level, an innovative immunocapture RT-LAMP (IC-RT-LAMP) and immunocapture reverse transcription (IC/RT-PCR) protocol on the basis of Potato Leafroll virus (PLRV) genome were used and optimized. In this regard, all six IC-RT-LAMP primers (i.e. F3, B3, FIP, BIP, LF and LB) together with IC/RT-PCR primers were designed on the basis of the highly conserved sequence (ORF3) of coat protein gene (GenBank accession number: U73777) of PLRV genome. Even though DAS-ELISA, IC/RT-PCR and IC-RT-LAMP assays could successfully detect positive infected plant samples, considering the time, safety, sensitivity, cost and simplicity, the last one was overall superior. Meanwhile, among five different visual dyes to accurately detect IC-RT-LAMP products, both hydroxynaphthol blue and GeneFinder™ could produce long stable colour change and brightness in a close tube-based approach to prevent cross-contamination risk, concluded eventually as the best ones. Altogether, as IC-RT-LAMP is sensitive, cost-effective, fairly user friendly and also can generate more accurate results than previous diagnostic procedures, we accordingly propose this colorimetric assay as a highly reliable alternative viral recognition system regarding PLRV recognition and probably other viral-based diseases.