Metabolic Response of Peach Fruit to Invasive Brown Marmorated Stink Bug (Halyomorpha halys Stål.)'s Infestation.

The brown marmorated stink bug (BMSB; Halyomorpha halys Stål.) is a highly destructive and polyphagous invasive pest that poses a serious threat to more than a hundred reported host plants. In the current study, the metabolic response of peach fruit of two cultivars-'Maria Marta' and 'Redhaven'-to BMSB infestation was studied using high-performance liquid chromatography (HPLC) and mass spectrometry (MS). In general, a strong phenolic response to BMSB infestation in peach flesh in the injury zone was observed, with flavanol content increasing by 2.4-fold, hydroxycinnamic acid content by 5.0-fold, flavonol content by 3.2-fold, flavanone content by 11.3-fold, and dihydrochalcones content by 3.2-fold compared with the undamaged tissue in the cultivar 'Maria Marta'. The phenolic response in the 'Redhaven' cultivar was even stronger. Consequently, the total phenolic content in the injured flesh also increased, 3.3-fold in 'Maria Marta' and 6.9-fold in 'Redhaven', compared with the uninjured flesh. Infestation with BMSB induced the synthesis of cyanidin-3-glucoside, which is not normally present in peach flesh. In comparison, the phenolic response was lower in peach peel, especially in the cultivar 'Maria Marta'. The study showed that both peach cultivars reacted to BMSB infestation with an increase in phenolic content in the peach flesh, but in a limited area of injury.

The Darker the Better: Identification of Chemotype Profile in Soroses of Local and Introduced Mulberry Varieties with Respect to the Colour Type.

Mulberries are the "essence of the past", the so-called Proust effect, for the inhabitants of the sericultural regions who enthusiastically remember feeding silkworms with mulberry leaves and picking the different coloured fruits that were their favourite sweets in childhood. To determine the chemistry behind the colour and taste of mulberry soroses, the main metabolites of the local and introduced varieties were studied. The soroses were classified into five different colour types and the size parameters were determined. The main sugars identified were glucose and fructose, while the predominant organic acids were citric and malic acids, which were highest in the darker varieties, and fumaric and tartaric acids, which were highest in the lighter varieties. A total of 42 phenolic compounds were identified. The predominant phenolic acid was chlorogenic acid, followed by other caffeoylquinic acids and coumaroylquinic acids. The predominant anthocyanins were cyanidin-3-glucoside and cyanidin-3-rutinoside. According to PCA analysis, the colour types showed a clear chemotype character. The sweet taste of the yellowish-white soroses was defined by 49% fructose, followed by 45% glucose and 6% organic acids. The sour character of the black genotypes was characterised by a lower sugar and higher (11%) organic acid content. The colour- and species-dependent effect was observed in the proportion of caffeoylquinic acids and quercetin glycosides, which decreased with increasing colour intensity from 60% of the total to 7%, and from 17% to 1%, respectively. An upward trend was observed for flavanols (5% to 29%) and anthocyanins, which accounted for 62% of the total phenolics in black varieties. This article gives an insight into the metabolite composition of mulberry soroses as the sweets of choice between light and sweet and dark and sour.

Breeding of Modern Rose Cultivars Decreases the Content of Important Biochemical Compounds in Rose Hips.

This study aimed to determine the content and composition of bioactive compounds in autochthonous rose hips (R. pendulina, R. spinosissima, and R. gallica) and to compare them with the content of bioactive compounds in some cultivars ('Harstad', 'Bourgogne', 'Mount Everest', 'Poppius', 'Fruhlingsduft', 'Single Cherry', 'Fruhlingsmorgen', 'Violacea', and 'Splendens') derived from these main species. Due to insufficient information on how bioactive compound content changes when crossing roses, this study also sought to ascertain whether modern rose hip cultivars are still a sufficiently rich source of bioactive compounds and could, therefore, be potentially used as a functional food. All material was collected in the Arboretum Volcji Potok (Slovenia). The ascorbic acid content was highest in the 'Harstad' cultivar (12.79 g/kg FW), and the total organic acid content varied from 1.57 g/kg FW (R. spinosissima) to 34.39 g/kg FW ('Harstad'). Of all the carotenoids analyzed, only lycopene and ß-carotene were present in all the samples. The total carotenoid content was highest in the 'Fruhlingsmorgen' cultivar (100.84 mg/kg FW), derived from R. spinosissima, and lowest in the main species, R. spinosissima (9.26 mg/kg FW). It can be concluded, therefore, that the content of bioactive compounds in rose hips of modern cultivars is generally lower than in rose hips of old cultivars and original species included in this study. The research results confirm that modern breeding strategies are mainly focused on goals such as abundant flowering and resistance to diseases and pests and not so much on the content of bioactive compounds.

Changes in the Fruit Quality Parameters of Medlar Fruit (Mespilus germanica L.) after Heat Treatment, Storage, Freezing or Hoarfrost.

The present study deals with the comparison of traditional fruit processing methods on medlar fruits and their effects on sugar content, organic acids, and phenolic composition in the medlar fruit variety 'Domaca nesplja'. The study aimed to analyze which processing methods can be used to make technologically mature medlar fruits that are not yet suitable for consumption edible and to maintain their good chemical quality. The two major sugars in medlars are fructose (59.30 g/kg FW) and glucose (54.43 g/kg FW), and the most abundant organic acids present are malic (8.44 g/kg FW) and quinic acid (8.77 g/kg FW). A total of 38 different phenolic compounds were identified in the medlar fruits: 13 phenolic acids, 9 flavanols, 1 flavone, 3 flavanones, and 12 flavonol glycosides. To explicate: phenolic acids (532.85 mg/kg FW) and flavanols (375.21 mg/kg FW) predominated; neochlorogenic acid had the highest content among phenolic acids; and procyanidins were the most abundant flavanols. The analysis observed statistical differences in metabolite content amongst fruits treated differently (technologically ripe fruits (harvested from the three fruits), edible fruits (technologically ripe fruits stored at 8 °C for 25 days), fruits exposed to the hoarfrost (temperature -1 °C to -4 °C), fruits heated at 60 °C (3 h), and frozen fruits (at -20 °C for 2 months). The lowest levels of fructose (191.77-195.1 g/kg DW) and sorbitol (29.35-31.3 g/kg DW) were detected in the heated and edible fruits. Edible fruits had a 30% lower content of organic acids than technologically ripe fruits and a five times lower content of flavanols, whereas flavonols had an 18.7 times lower content of phenolic acids than technologically ripe fruits. Heating the fruits to 60 °C resulted in a 40% increase in total phenolic compounds in medlars. The results of the study indicate that exposure of medlar fruit to hoarfrost does not significantly affect the chemical quality of the fruit and only minimally alters the composition of sugars, acids, and phenolic compounds. The processing of medlar fruit with hoarfrost, therefore, remains the most suitable method of fruit bletting.

Detailed Metabolic Characterization of Flowers and Hips of Rosa gallica L. Grown in Open Nature.

Our research aimed to investigate the primary and secondary metabolites of rosehips and petals of R. gallica in comparison with R. subcanina. R. gallica was chosen because it is still unexplored in terms of various bioactive substances and is strongly present in Slovenia. Given that roses are generally very variable and unstudied, our research will contribute to greater transparency and knowledge of the bioactive composition of rosehips and petals. We found a strong positive correlation between the total content of phenolics and ascorbic acid, between the total content of organic acids and the total content of carotenoids, and between the total content of sugars and the total content of organic acids. Hips of R. gallica contained higher amounts of sugars, ascorbic acid, and carotenoids than R. subcanina. Based on the composition of phenolic compounds in the petals, it is possible to distinguish between the two species. Among all the phenolic compounds in the petals, both genotypes are richest in gallotannins, followed by flavonols. Among anthocyanins, cyanidin-3-glucoside was determined, the content of which was also higher in R. gallica. It can be concluded that the studied hips had an extremely low sugar content and, consequently, an extremely high organic acid content. The content of carotenoids in hips was in the lower range of the average content compared to data from the literature. By optimizing the harvesting time, we could obtain a higher content of carotenoids, which could potentially be used for industrial purposes. However, we found that the analyzed petals were a rich source of phenolic compounds, which benefit the human body and could be potentially used in the food and cosmetic industries.

Invasive Alien Species as a Potential Source of Phytopharmaceuticals: Phenolic Composition and Antimicrobial and Cytotoxic Activity of Robinia pseudoacacia L. Leaf and Flower Extracts.

Black locust (Robinia pseudoacacia L.), an invasive tree in Europe, commonly known for its negative impact on biodiversity, is a rich source of phenolic compounds recognized in traditional medicine. Since the metabolite profile depends on the environment and climate, this study aimed to provide the first LC-MS phytochemical screening of the black locust from the Istria region (Croatia). The compounds were extracted from leaves and flowers with 70% ethanol and 80% methanol. Total phenolics (TP) and flavonoids (TF), as well as antioxidant capacity (AC) measured by ABTS (17.49-146.41 mg TE/g DW), DPPH (24.67-118.49 mg TE/g DW), and FRAP (7.38-77.53 mg TE/g DW) assays, were higher in leaf than in flower extracts. Higher TP and total non-flavonoid (TNF) values were displayed in ethanolic than in methanolic extracts. In total, 64 compounds were identified, of which flavonols (20) and hydroxycinnamic acid derivatives (15) were the most represented. Flavanols such as catechin dominated in leaf extracts, followed by flavonols, with kaempferol glucuronyl rhamnosyl hexosides as the main compound, respectively. Flower extracts had the highest share of flavones, followed by ellagitannins, with luteolin dirhamnosyl hexosides and vescalagin, respectively, being predominant. The extracts had good quorum sensing, biofilm formation prevention, and eradicating capacity. The results provided new insights into the phytochemical properties of R. pseudoacacia as the first step toward its potential pharmaceutical use.

Black Queens of Fruits: Chemical Composition of Blackberry (Rubus subg. rubus Watson) and Black Currant (Ribes nigrum L.) Cultivars Selected in Serbia.

Black fruits, especially blackberries and black currants, are highly appreciated by consumers due to their nutraceutical properties, which have reported health benefits. This study aimed to assess the fruit quality of the blackberry (cv. 'Cacanska Bestrna') and black currant (cv. 'Cacanska Crna') created at the Fruit Research Institute, Cacak (Republic of Serbia) by evaluating basic quality parameters (fruit weight, soluble solids), and content of primary (sugars and organic acids) and secondary (phenolic compounds) metabolites. Additionally, the study examined the quality of frozen and long-term frozen storage (6, 9, and 12 months). The results showed that 'Cacanska Crna' contained a high level of total soluble solids (15.23°Bx), sugars (93.06 mg g-1), and a high index of sweetness (159.48) as well as a high content of acids (34.35 mg g-1) in the fruit. Both species were found to contain forty-seven phenolic compounds, including phenolic acids, flavanols, and anthocyanins, analyzed using high performance liquid chromatography-mass spectrometry (HPLC-MS). 'Cacanska Bestrna' had the highest amounts of phenolic acids (66.85 mg 100 g-1) and flavanols (53.99 mg 100 g-1), whereas 'Cacanska Crna' showed the highest levels of flavonols (8.57 mg 100 g-1) and anthocyanins (139.11 mg 100 g-1). Furthermore, the study revealed that anthocyanins were the most abundant phenolic group in both blackberries and black currants, and their levels remained constant during frozen storage. The sugar content in both species remained unchanged, while the acid concentration increased over time in blackberries but remained consistent in black currants. Overall, the findings highlight the superior phenolic content, particularly anthocyanins, of the cultivars with black fruits selected in Serbia. These cultivars have great commercial potential for the fresh market and processing. Moreover, the study suggests that frozen storage is an effective method for preserving their quality and beneficial properties.

Phenolic Compounds of Rose Hips of Some Rosa Species and Their Hybrids Native Grown in the South-West of Slovenia during a Two-Year Period (2020-2021).

The genus Rosa is very extensive and variable, so it remains very unpredictable and uninvestigated. This also holds true for values of secondary metabolites in rose hips, which are important for several purposes (human diet, protection of plants against pests, etc.). The aim of our study was to determine the content of phenolic compounds in the hips of R. × R. glauca, R. corymbifera, R. gallica and R. subcanina, which grow wild in nature in southwestern Slovenia. We examined the content of phenolic compounds in different parts of rose hips, in the flesh with skin and in the seeds, depending on the individual species, over a period of two years, 2020 and 2021. We also considered the influence of environmental conditions on the content of the mentioned compounds. In both years, the content of phenolic compounds was higher in the flesh with skin than in the seeds. Considering the total content of phenolic compounds in the flesh with skin, R. gallica stands out (15,767.21 mg/kg FW), but the hips of this species accumulated the lowest number of different phenolic compounds. The lowest content of total phenolic compounds (TPC) was found in R. corymbifera, in the year 2021 (3501.38 mg/kg FW). The content of TPC (in both observed years) in the seeds varied between 1263.08 mg/kg FW (R. subcanina) and 3247.89 mg/kg FW (R. × R. glauca). Among the anthocyanins, cyanidin-3-glucoside was determined, which was predominant in R. gallica (28.78 mg/kg FW), and at least was determined in R. subcanina (1.13 mg/kg FW). When comparing the two years of the period (2020-2021), we found that 2021 was more favorable for the formation of phenolic compounds in the seeds, but 2020 in the flesh with skin.

Preventive and Curative Effects of Salicylic and Methyl Salicylic Acid Having Antifungal Potential against Monilinia laxa and the Development of Phenolic Response in Apple Peel.

The effects of salicylic acid (SA) and one of its better-known derivatives-methyl salicylic acid (MeSA)-on the infection of apple fruits with the fungus Monilinia laxa, which causes brown rot, were investigated. Since research to date has focused on preventive effects, we also focused on the curative use of SA and MeSA. Curative use of SA and MeSA slowed the progression of the infection. In contrast, preventive use was generally unsuccessful. HPLC-MS was used to analyze the content of phenolic compounds in apple peels in healthy and boundary peel tissues around lesions. The boundary tissue around the lesions of untreated infected apple peel had up to 2.2-times higher content of total analyzed phenolics (TAPs) than that in the control. Flavanols, hydroxycinnamic acids and dihydrochalcones were also higher in the boundary tissue. During the curative treatment with salicylates, the ratio of TAP content between healthy and boundary tissue was lower (SA up to 1.2-times higher and MeSA up to 1.3-times higher content of TAPs in boundary compared to those in healthy tissue) at the expense of also increasing the content in healthy tissues. The results confirm that salicylates and infection with the fungus M. laxa cause an increased content of phenolic compounds. Curative use of salicylates has a greater potential than preventive use in infection control.

Bioprocessed Wholegrain Spelt Flour Improves the Quality and Physicochemical Characteristics of Wheat Bread.

In the present study, the partial substitution of common white wheat flour for a bread recipe with variously bioprocessed wholegrain spelt was investigated. The addition of 1% and pasteurised 5% "germinated + enzymatic treated" spelt flour to wheat flour significantly improved the specific volume of the bread, but their texture profile analysis and sensory evaluation were not satisfactory. A higher percentage of added bioprocessed spelt flour darkened the colour of the bread. Breads with the addition of more than 5% of bioprocessed spelt flour were unacceptable in terms of quality and sensory parameters. The highest extractable and bound individual phenolics were found in breads with 5% "germinated + fermented" spelt flour (GFB5) and 5% pasteurised "germinated + enzymatic treated" spelt flour (GEB5P). A strong positive correlation was determined between trans-ferulic acid and TPC and DPPH• radical scavenging activity. The GEB5P bread showed the highest increase in extractable and bound trans-ferulic acid content, by 320% and 137%, respectively, compared to the control bread. Principal component analysis showed differences between the control bread and enriched breads in terms of their quality, sensory and nutritional properties. Breads with 2.5% and 5% "germinated + fermented" spelt flour had the most acceptable rheological, technological and sensory characteristics, in addition to a substantial improvement in their antioxidant content.

Biochemical Characterization of Black and Green Mutant Elderberry during Fruit Ripening.

The content of sugars, organic acids, phenolic compounds and selected enzyme activities in the anthocyanin pathway were analyzed in NIGRA (Sambucus nigra var. nigra-black fruits) and VIRIDIS (S. nigra var. viridis-green fruits) fruits over four stages of ripening. The share of glucose and fructose in green fruits was higher than in colored fruits, and the sugar content increased significantly until the third developmental stage. Ripe NIGRA berries had 47% flavonol glycosides, 34% anthocyanins, 3% hydroxycinnamic acids and 14% flavanols, whereas the major phenolic group in the VIRIDIS fruits, making up 88% of the total analyzed polyphenols, was flavonols. NIGRA fruits were rich in anthocyanins (6020 µg g-1 FW), showing strong activation of the late anthocyanin pathway (dihydroflavonol 4-reductase, anthocyanidin synthase). In both color types, phenylalanine ammonia lyase and chalcone synthase/chalcone isomerase activities were highest in the first stage and decreased during ripening. In VIRIDIS fruit, no anthocyanins and only one flavanol (procyanidin dimer) were found. This was most likely caused by a lack of induction of the late anthocyanin pathway in the last period of fruit ripening. The VIRIDIS genotype may be useful in studying the regulatory structures of anthocyanin biosynthesis and the contribution of distinct flavonoid classes to the health benefits of elderberries.

Characterization of the Slovene Autochthonous Rose Hybrid Rosa pendulina × spinosissima (Rosa reversa Waldst. and Kit) Using Biochemical Patterns of the Plant Blossoms.

The Rosa genus is characterized by great variability and, consequently, they easily hybridize. The petals of R. pendulina, R. spinosissima and their hybrid Rosa pendulina × spinosissima, collected in western Slovenia, were included in the research. We performed morphometric analysis using keys to determine roses and genetic analysis to determine the genome size. The phenolic compound content in petals of all rose flowers was measured by liquid chromatography and mass spectrometry (HPLC-MS). Using flow cytometry, we could confirm the native hybridization process due to the amount of 2C DNA. The value of R. pendulina was 1.71 pg, of R. spinosissima 1.60 pg and of the hybrid 1.62 pg. The value for the hybrid was close to values measured for parent plants and, at the same time, those values of parent plants significantly differed from each other. Our results showed that the content of phenolic compounds in petals decreased after crossing. We found that the highest total phenolic content (178.9 g/kg FW) was measured in R. spinossisima, the lowest content was analyzed for the hybrid (84.36 g/kg FW) and the content for R. pendulina was between these two values (110.58 g/kg FW). The content of flavanols and flavonols was lowest in the hybrid petals, whereas the content of anthocyanins was highest in the hybrid petals.

Changes in the Bioaccessibility of Antioxidants after Simulated In Vitro Digestion of Bioprocessed Spelt-Enhanced Wheat Bread.

The aim of the study was to determine whether the partial replacement of wheat flour with bioprocessed spelt flour contributes to a higher bioaccessibility of the antioxidants in bread. The results showed that the type and amount of bioprocessed spelt flour in a bread recipe has a major impact on the extractable and bound TPC, the content of individual phenolics, their antioxidant activity, and their bioaccessibility as determined by in vitro digestion. Extractable p-coumaric and trans-ferulic acids in breads decreased after digestion, while extractable cis-ferulic and p-hydroxybenzoic acids increased. The bioaccessibility of TPC in the control bread (100% wheat flour), and in bread enriched with 5% "germinated + fermented" spelt flour (GFB5), did not differ. However, the digested GFB5 bread contained 5.2-times more extractable, and 1.3-times more bound, trans-ferulic acid than the digested control bread. trans-Ferulic acid showed the lowest bioaccessibility, up to 2.8%. In GFB2.5 and GFB5 breads, the bioaccessibility of p-coumaric, trans-ferulic, and cis-ferulic acids was higher than in other digested breads. PCA visualized the difference between the undigested and digested breads. The incorporation of germinated and fermented, or germinated and enzymatic, treated spelt flour in a white bread recipe could be an attractive way of providing consumers with nutritionally interesting foods.

The Effects of the Cultivar and Environment on the Phenolic Contents of Hazelnut Kernels.

Different climatic conditions are known to affect the synthesis of primary and secondary metabolites. Therefore, the phenolic contents in new growing areas could affect the quality and flavor of hazelnuts. The aim of this study was to determine the variability of the phenolic contents of the kernels in different commercial hazelnut cultivars depending on their growing area. Five cultivars ('Tonda Gentile delle Langhe', 'Merveille de Bollwiller', 'Pauetet', 'Tonda di Giffoni', and 'Barcelona' (syn. 'Fertile de Coutard')) grown in different European collection orchards were included in the study. High-performance liquid chromatography coupled with mass spectrometry was used to identify and quantify the phenolic compounds. Thirteen phenols were identified in the hazelnut kernels, including 7 flavanols, 2 hydroxybenzoic acids, 3 flavonols, and one dihydrochalcone. Catechin and procyanidin dimers were the main phenolic compounds found in the hazelnut kernels. The highest contents of catechin and total flavanols were determined in cultivars cultivated in Spain and northern Italy, and the lowest in Slovenia and France. Flavanols were the major phenolic groups independent of the place of cultivation, as they accounted for more than 50% of all phenolic compounds identified. The flavanols were followed by hydroxybenzoic acids, flavonols, and dihydrochalcones. Higher contents of flavanols and flavonols were found in kernels from areas characterized by higher natural irradiation, which stimulates their accumulation. The contents of hydroxybenzoic acids correlated with altitude, which stimulated phenolic acid synthesis. A negative correlation was observed between the dihydrochalcone content and annual rainfall, probably due to hydric stress.

HPLC-DAD-MS Identification and Quantification of Phenolic Components in Japanese Knotweed and American Pokeweed Extracts and Their Phytotoxic Effect on Seed Germination.

We performed a detailed HPLC-MSn analysis of the phenolic compounds from the extracts of two invasive alien plant species (IAPS): Japanese knotweed (Fallopia japonica (Houtt.) Ronse Decr.) and American pokeweed (Phytolacca americana L.). The major phenolic groups were hydroxycinnamic acids and flavanols in Japanese knotweed (J. knotweed) and flavonols, hydroxycinnamic acids, and stilbenes in American pokeweed (A. pokeweed). We investigated the influence of solvent type and extraction time on the extraction efficiency of the phenolic compounds. The solvent 80% methanol had a higher polyphenolic extraction efficiency than water, since 14.5 times more flavonols and 2.3 times more stilbenes were extracted from J. knotweed and 5.2 times more flavonols and 2.6 times more stilbenes were extracted from A. pokeweed. In contrast, with water, we obtained a 52% higher hydroxycinnamic acids (HCA) content from J. knotweed. Hydroxycinnamic acids were best extracted in water after 24 h, flavanols after 12 h, stilbenes between 12 and 24 h, and flavonol glycosides after 48 h of extraction. We also tested the allelopathic effect of the aqueous extract of A. pokeweed and J. knotweed on seed germination and shoot and root growth of perennial ryegrass. The results showed that the water extract of J. knotweed resulted in 38 to 48% lower seed germination of perennial ryegrass, and the extract of A. pokeweed resulted in 83 to 90% lower seed germination. The phytotoxic effect of the extract of J. knotweed and A. pokeweed was also reflected in a characteristic reduced growth of shoots and roots of perennial ryegrass. The phytotoxic action of IAPS could also be applied for beneficial purposes, since this would be an effective strategy for their control and a reduction of their spread in the environment.

Simulated Gastrointestinal Digestion of Bioprocessed Spelt Seeds: Bioaccessibility and Bioactivity of Phenolics.

The goal of this research was to evaluate the impact of different bioprocessing techniques on improved bioaccessibility of phenolics from spelt seeds. Despite the negative influence of gastrointestinal digestion, fermentation of germinated seeds significantly increased the bioaccessibility of total phenolics and their antioxidant activity compared to digested raw seeds. Enzymatic treated fermented seeds showed the highest relative bioaccessibility of p-coumaric and trans-ferulic acids, while their absolute contents were significantly higher in "germinated + fermented" seeds. Our research suggests that pretreatment of spelt seeds with hydrolytic enzymes improves access of fermenting microorganisms to structural elements, resulting in an increased content of extractable and bound trans-ferulic acid. Significantly higher biostability of phenolics was observed in raw seeds. Some major quality changes in the composition of extracts were observed under simulated in vitro digestion, since antioxidants of the same extract showed a different relative decrease in DPPH• and ABTS•+ scavenging activities compared to the raw seeds or their corresponding undigested counterparts. It is therefore important to increase the content of extractable antioxidants in seeds by bioprocessing, since they are strongly diminished during digestion.

Improving accessibility and bioactivity of raw, germinated and enzymatic-treated spelt (Triticum spelta L.) seed antioxidants by fermentation.

Lactic acid, alcoholic, combined and spontaneous fermentation of raw, germinated and enzymatic-treated spelt seeds significantly improved the content of extractable and bound phenolics and considerably increased the extractable:bound ratio, and therefore positively affected the accessibility of the spelt antioxidants. The highest extractable and bound individual phenolic contents and in vitro antioxidant activities of extracts were obtained following fermentation of germinated spelt seeds with Saccharomyces cerevisiae, while for enzymatic-treated seeds, Lactobacillus plantarum (alone or with S. cerevisiae) was the most effective. For extractable phenolics, trans-ferulic acid increased the most in yeast-fermented germinated seeds (2922%); for bound phenolics, cis-ferulic acid showed the greatest relative increase in yeast-fermented raw spelt seeds (466%). Spontaneous fermentation of germinated and enzymatic-treated samples decreased intracellular oxidation most effectively, probably due to apigenin derivatives. Cellular uptake of bound hydroxycinnamic acids was significantly higher than that of extractable hydroxycinnamic acids; however, the latter were more efficient in vivo antioxidants.

LC-DAD-MS Phenolic Characterisation of Six Invasive Plant Species in Croatia and Determination of Their Antimicrobial and Cytotoxic Activity.

Invasive plants' phytochemicals are important for their invasiveness, enabling them to spread in new environments. However, these chemicals could offer many pharmaceutical compounds or active ingredients for herbal preparations. This study provides the first LC-MS phytochemical screening of six invasive alien plant species (IAPS) in the Istria region (Croatia): Ailanthus altissima, Ambrosia artemisiifolia, Conyza canadensis, Dittrichia viscosa, Erigeron annuus, and Xanthium strumarium. The study aims to identify and quantify the phenolic content of their leaf extracts and assess their antimicrobial and cytotoxic potential. A total of 32 species-specific compounds were recorded. Neochlorogenic, chlorogenic, and 5-p-coumaroylquinic acids, quercetin-3-glucoside, and kaempferol hexoside were detected in all the tested IAPS. Hydroxycinnamic acid derivatives were the main components in all the tested IAPS, except in E. annuus, where flavanones dominated with a share of 70%. X. strumarium extract had the best activity against the tested bacteria, with an average MIC value of 0.11 mg/mL, while A. altissima and X. strumarium extracts had the best activity against the tested fungi, with an average MIC value of 0.21 mg/mL in both cases. All the plant extracts studied, except X. strumarium, were less cytotoxic than the positive control. The results provided additional information on the phytochemical properties of IAPS and their potential for use as antimicrobial agents.

Biopotential of Underutilized Rosaceae Inflorescences: LC-DAD-MS Phytochemical Profiles Associated with Antioxidant, Antidiabetic, Anti-Inflammatory and Antiproliferative Activity In Vitro.

The aim of this work was to assess the biopotential of the young inflorescence tissues of Prunus, Malus and Chaenomeles in order to evaluate the possibility of their application in the food industry, and to provide a polyphenolic fingerprint for their quality control. The contents of different bioactive compounds and their antioxidant capacities were spectrophotometrically measured, the main phenolic compounds were identified and quantified using LC-DAD-MS, the antidiabetic potential was determined using α-amylase and α-glucosidase inhibition assays, the anti-inflammatory potential was determined using a 5-lipoxygenase inhibition assay, and the cytotoxicity was determined by MTT assay. Using one-way ANOVA, principal component analysis, hierarchical clustering and Pearson's correlation coefficient, the relations between the samples, and between the samples and the measured parameters, were revealed. In total, 77 compounds were identified. The concentration of sugars was low in M. purpurea, at 1.56 ± 0.08 mg/g DW. The most effective sample in the inhibition of antidiabetic enzymes and anti-inflammatory 5-lipoxygenase was C. japonica. The inhibition of α-glucosidase was strongly positively correlated with the total and condensed tannins, procyanidin dimers and procyanidin tetramer, and was very strongly correlated with chlorogenic acid. In α-amylase inhibition, C. japonica and P. serrulata 'Kiku Shidare Zakura' were equally efficient to the standard inhibitor, maltose. The most effective in the growth and proliferation inhibition of HepG2, HCT116 and HaCaT cells was P. avium. The results suggest Prunus, Malus and Chaenomeles inflorescences as functional food ingredients.

The Phenolic Profile of Sweet Cherry Fruits Influenced by Cultivar/Rootstock Combination.

The influence of three cultivars ('Carmen', 'Kordia' and 'Regina') grafted on six rootstocks (Mahaleb, 'Colt', 'Oblacinska', 'M × M 14', 'Gisela 5' and 'Gisela 6') on the phenolic profile of sweet cherry fruits was studied during a two-year period. All the individual phenolic compounds were detected using high-pressure liquid chromatography with diode-array detection coupled with mass spectrometry (HPLC-DAD-MSn). In all the examined samples, 54 compounds were identified and divided into five phenolic classes: anthocyanins (4 compounds), flavonols (7), flavanols (11), flavanones (4), and hydroxycinnamic acids (28). Anthocyanins (58%) and hydroxycinnamic acids (31%) showed the greatest amounts in all the examined fruit samples. PCA analysis revealed that among the cultivars, 'Kordia' showed the highest phenolic content. Regarding rootstocks, the lowest values of the most important phenolic compounds were obtained in fruits from trees grafted onto the seedling rootstock Mahaleb. Among the clonal rootstocks, the vigorous 'Colt' and dwarf 'Gisela 5' promoted the highest values of the evaluated phenolic compounds in the cultivars 'Kordia' and 'Carmen', while the dwarf 'Oblacinska' and semi-vigorous 'M × M 14' induced the highest values in the cultivar 'Regina'. By evaluating the influence of cultivars and rootstocks on the phenolic content in fruit, it has been proven that the cultivar has the most significant influence. However, the rootstock also influences the content of a large number of phenolic compounds. The selection of an adequate cultivar/rootstock combination can also be a powerful tool for improving the phenolic content in fruits, and consequently the nutritional value of sweet cherry fruits.