Identification and characterisation of CYP75A31, a new flavonoid 3'5'-hydroxylase, isolated from Solanum lycopersicum

<p>Abstract</p> <p>Background</p> <p>Understanding the regulation of the flavonoid pathway is important for maximising the nutritional value of crop plants and possibly enhancing their resistance towards pathogens. The flavonoid 3'5'-hydroxylase (F3'5'H) enzyme functions at an important branch point between flavonol and anthocyanin synthesis, as is evident from studies in petunia (<it>Petunia hybrida</it>), and potato (<it>Solanum tuberosum</it>). The present work involves the identification and characterisation of a <it>F3'5'H </it>gene from tomato (<it>Solanum lycopersicum</it>), and the examination of its putative role in flavonoid metabolism.</p> <p>Results</p> <p>The cloned and sequenced tomato <it>F3'5'H </it>gene was named <it>CYP75A31</it>. The gene was inserted into the <it>pYeDP60 </it>expression vector and the corresponding protein produced in yeast for functional characterisation. Several putative substrates for F3'5'H were tested <it>in vitro </it>using enzyme assays on microsome preparations. The results showed that two hydroxylation steps occurred. Expression of the <it>CYP75A31 </it>gene was also tested <it>in vivo</it>, in various parts of the vegetative tomato plant, along with other key genes of the flavonoid pathway using real-time PCR. A clear response to nitrogen depletion was shown for <it>CYP75A31 </it>and all other genes tested. The content of rutin and kaempferol-3-rutinoside was found to increase as a response to nitrogen depletion in most parts of the plant, however the growth conditions used in this study did not lead to accumulation of anthocyanins.</p> <p>Conclusions</p> <p><it>CYP75A31 </it>(NCBI accession number GQ904194), encodes a flavonoid 3'5'-hydroxylase, which accepts flavones, flavanones, dihydroflavonols and flavonols as substrates. The expression of the <it>CYP75A31 </it>gene was found to increase in response to nitrogen deprivation, in accordance with other genes in the phenylpropanoid pathway, as expected for a gene involved in flavonoid metabolism.</p

A novel bicyclic lactone and other polyphenols from the commercially important vegetable Anthriscus cerefolium

Garden chervil, Anthriscus cerefolium (L.) Hoffm. is an important herb commonly applied in Norwegian large-scale commercial kitchens. This species is a highly enriched source of phenolics, containing 1260 mg gallic acid equivalents (GAE) 100–1 g DM, however, the individual phenolic compounds have been scarcely characterized. Here we report on the qualitative and quantitative content of phenolics in garden chervil. The structure of the main phenolic compound was elucidated to be the previously undescribed compound 1,3-dicaffeoyl-5-malonyl-δ-quinide (1) by means of 1D- and 2D NMR and high-resolution mass spectrometry. The known flavones apigenin 7-O-β-(2″-apiofuranosylglucopyranoside) (= apiin) (2), apigenin 7-(2″-apiosyl-6″-malonylglucoside) (3) and luteolin 7-glucoside (4) were also identified. Compound 3 is reported for the first time from this plant species. The main phenolic compound, 1,3-dicaffeoyl-5-malonyl-δ-quinide, exhibited moderate cytotoxicity towards acute monocytic leukaemia cells (MOLM-13) and rat kidney epithelial cells (NRK) with EC50 between 400 and 600 µM.publishedVersio

Evaluation of Antioxidant Compounds and Total Sugar Content in a Nectarine [Prunus persica (L.) Batsch] Progeny

Epidemiological studies suggest that consumption of fruit rich in phenolic compounds is associated with health-protective effects due to their antioxidant properties. For these reasons quality evaluation has become an important issue in fruit industry and in breeding programs. Phytochemical traits such as total phenolics, flavonoids, anthocyanins, L-ascorbic acid, sugar content and relative antioxidant capacity (RAC) were analyzed over four years in flesh fruit of an F1 population “Venus” × “Big Top” nectarines. Other traits such as harvesting date, yield, fruit weight, firmness, soluble solids concentration (SSC), pH, titratable acidity (TA) and ripening index (RI) were also determined in the progeny. Results showed high variability among genotypes for all analyzed traits. Total phenolics and flavonoids showed significant positive correlations with RAC implying that both are important antioxidant bioactive compounds in peaches. We found genotypes with enhanced antioxidant capacity and a better performance than progenitors, and in consequence the best marketability

Variable response of three Trifolium repens ecotypes to soil flooding by seawater.

BACKGROUND AND AIMS: Despite concerns about the impact of rising sea levels and storm surge events on coastal ecosystems, there is remarkably little information on the response of terrestrial coastal plant species to seawater inundation. The aim of this study was to elucidate responses of a glycophyte (white clover, Trifolium repens) to short-duration soil flooding by seawater and recovery following leaching of salts. METHODS: Using plants cultivated from parent ecotypes collected from a natural soil salinity gradient, the impact of short-duration seawater soil flooding (8 or 24 h) on short-term changes in leaf salt ion and organic solute concentrations was examined, together with longer term impacts on plant growth (stolon elongation) and flowering. KEY RESULTS: There was substantial Cl(-) and Na(+) accumulation in leaves, especially for plants subjected to 24 h soil flooding with seawater, but no consistent variation linked to parent plant provenance. Proline and sucrose concentrations also increased in plants following seawater flooding of the soil. Plant growth and flowering were reduced by longer soil immersion times (seawater flooding followed by drainage and freshwater inputs), but plants originating from more saline soil responded less negatively than those from lower salinity soil. CONCLUSIONS: The accumulation of proline and sucrose indicates a potential for solute accumulation as a response to the osmotic imbalance caused by salt ions, while variation in growth and flowering responses between ecotypes points to a natural adaptive capacity for tolerance of short-duration seawater soil flooding in T. repens. Consequently, it is suggested that selection for tolerant ecotypes is possible should the predicted increase in frequency of storm surge flooding events occur

Reduction of inositol (1,4,5)–trisphosphate affects the overall phosphoinositol pathway and leads to modifications in light signalling and secondary metabolism in tomato plants

The phosphoinositol pathway is one of the major eukaryotic signalling pathways. The metabolite of the phosphoinositol pathway, inositol- (1,4,5) trisphosphate (InsP3), is a regulator of plant responses to a wide variety of stresses, including light, drought, cold, and salinity. It was found that the expression of InsP 5-ptase, the enzyme that hydrolyses InsP3, also dramatically affects the levels of inositol phosphate metabolites and the secondary metabolites in transgenic tomato plants. Tomato plants expressing InsP 5-ptase exhibited a reduction in the levels of several important inositol phosphates, including InsP1, InsP2, InsP3, and InsP4. Reduced levels of inositol phosphates accompanied an increase in the accumulation of phenylpropanoids (rutin, chlorogenic acid) and ascorbic acid (vitamin C) in the transgenic fruits of tomato plants. The enhanced accumulation of these metabolites in transgenic tomato plants was in direct correspondence with the observed up-regulation of the genes that express the key enzymes of ascorbic acid metabolism (myo-inositol oxygenase, MIOX; L-galactono-γ-lactone dehydrogenase, GLDH) and phenylpropanoid metabolism (chalcone synthase, CHS1; cinnamoyl-CoA shikimate/quinate transferase, HCT). To understand the molecular links between the activation of different branches of plant metabolism and InsP3 reduction in tomato fruits, the expression of transcription factors known to be involved in light signalling was analysed by real-time RT-PCR. The expression of LeHY5, SIMYB12, and LeELIP was found to be higher in fruits expressing InsP 5-ptase. These results suggest possible interconnections between phosphoinositol metabolism, light signalling, and secondary metabolism in plants. Our study also revealed the biotechnological potential for the genetic improvement of crop plants by the manipulation of the phosphoinositol pathway

Fruit-Surface Flavonoid Accumulation in Tomato Is Controlled by a SlMYB12-Regulated Transcriptional Network

The cuticle covering plants' aerial surfaces is a unique structure that plays a key role in organ development and protection against diverse stress conditions. A detailed analysis of the tomato colorless-peel y mutant was carried out in the framework of studying the outer surface of reproductive organs. The y mutant peel lacks the yellow flavonoid pigment naringenin chalcone, which has been suggested to influence the characteristics and function of the cuticular layer. Large-scale metabolic and transcript profiling revealed broad effects on both primary and secondary metabolism, related mostly to the biosynthesis of phenylpropanoids, particularly flavonoids. These were not restricted to the fruit or to a specific stage of its development and indicated that the y mutant phenotype is due to a mutation in a regulatory gene. Indeed, expression analyses specified three R2R3-MYB–type transcription factors that were significantly down-regulated in the y mutant fruit peel. One of these, SlMYB12, was mapped to the genomic region on tomato chromosome 1 previously shown to harbor the y mutation. Identification of an additional mutant allele that co-segregates with the colorless-peel trait, specific down-regulation of SlMYB12 and rescue of the y phenotype by overexpression of SlMYB12 on the mutant background, confirmed that a lesion in this regulator underlies the y phenotype. Hence, this work provides novel insight to the study of fleshy fruit cuticular structure and paves the way for the elucidation of the regulatory network that controls flavonoid accumulation in tomato fruit cuticle

Some food toxic for pets

According to world statistics, dogs and cats are the species that owners most frequently seek assistance with potential poisonings, accounting 95–98% of all reported animal cases. Exposures occur more commonly in the summer and in December that is associated with the holiday season. The majority (>90%) of animal poisonings are accidental and acute in nature and occur near or at the animal owner's home. Feeding human foodstuff to pets may also prove dangerous for their health

Dynamics of quercetin formation in onion (Allium cepa L.) during vegetation

Onion bulbs (Allium cepa L.) are good sources of flavonoids. The aim of this study was to analyse the changes in dynamics of quercetin formation in three varieties of onions (white, yellow, and red) during the vegetation period. Quercetin content was determined after acid hydrolysis (1.2 M HCl in 50% aqueous methanol) by high-performance liquid chromatography (HPLC). The content of total phenolics was determined using Folin-Ciocalteu reagent (FCR) according to LACHMAN and co-workers (2003). The content of polyphenols in onion ranged from 2893 to 6052 mg kg–1 and the content of quercetin ranged from 52.44 to 280.72 mg kg–1 in fresh matter. The highest content of polyphenols and quercetin was found in the red variety. According to statistical analysis the dynamic of quercetin formation in all cultivars had statistically moderate (P<0.05) increasing tendency. Increasing content of polyphenols was accompanied with slight increase of quercetin, but the differences remained insignificant (P<0.05)

Распространенность тяжелого раннего детского кариеса (S-ECC)

ДЕТИ РАННЕГО ВОЗРАСТАДОШКОЛЬНИКИКАРИЕС ЗУБОВ /ЭПИДСТОМАТОЛОГИЧЕСКИЕ ОБСЛЕДОВАНИЯЗУБ МОЛОЧНЫЙКАРИЕС ДЕТСКИЙ РАННИЙЭПИДЕМИОЛОГИЧЕСКИЕ ИССЛЕДОВАНИЯНАСЕЛЕНИЯ ГРУПП ИЗУЧЕНИ

Screen-printed electrochemical-based sensor for taxifolin determination in edible peanut oils

An electrochemical method for the determination of Taxifolin (Tx) in peanut oil samples using screen-printed electrodes (SPEs) is reported for the first time. The different electroanalytical parameters such as scan rate, supporting electrolyte, pH and accumulation step were optimized. Interference studies were also explored for the determination of Tx in presence of daidzein, quercetin, trans-resveratrol and caffeic acid. The method developed for the Tx determination exhibits a linear response in the concentration range from 0.05 to 1 μM and a limit of detection of 0.021 μM. The electrochemical detection method proposed provides a novel, quick, low-cost, reliable and portable method for the in-situ quality control for monitoring the protected designation of origin of the “Córdoba, Argentina, Peanuts”