Phytochemical, Nutritional and Mineral Content of Four Edible Flowers.

The growing interest in functional foods is driven by the exploration of new foods with positive health effects. Pleasant sensory features are essential for consumer acceptance. In this work, we investigated the composition of the bioactive compounds, antioxidant activity, and aroma profiles of four edible flowers: Cucurbita moschata Duchesne, Dianthus chinensis L., Fuchsia regia (Vand. ex Vell.) Munz., and Viola cornuta L. For the first time, we quantified the water-soluble group of B vitamins. Significant variations in the content of soluble sugars, vitamins, and secondary metabolites were observed. V. cornuta showed the highest concentration of vitamin C and carotenoids, while C. moschata had the highest content of vitamin B and flavonoids. F. regia stood out for its exceptionally high content of total phenolics, while D. chinensis surpassed the other flowers in soluble sugar content. The aroma profile analysis revealed a diverse array of volatile organic compounds, with each species having its own unique composition. C. moschata was characterized by p-dimethoxybenzene and D. chinensis by non-terpene compounds; F. regia displayed high amounts of decanal and nonanal, while V. cornuta was rich in myrcene and α-farnesene. These findings provide valuable insights into the secondary metabolites and aroma profiles of these flowers, enhancing our understanding of their bioactive compounds and potential health benefits.

Phytonutritional compounds and antioxidant activity of eight new edible flowers.

In the last years, special attention has been given to the nutritional properties of food, and the edible flowers (EFs) represent an uncommon fresh product to characterise, with the aim to explore new species with peculiar aesthetic and the aromatic features. This work investigated some nutritional properties of eight new EFs, such as Begonia boliviensis A.DC. (pink and white varieties), Dahlia pinnata Cav., Salvia farinacea Benth., S. × jamensis J.Compton, S. 'Purple Queen' Tulbaghia simmleri Beauv. 'Alba', T. violacea Harv. 'Alba', chosen for their different colours and corolla morphologies. Primary and secondary metabolites have been determined. The highest protein content was found in T. violacea 'Alba' and D. pinnata. The Salvia species showed the highest soluble sugar content. The two Tulbaghia species were notable for hexose and ascorbic acid content. D. pinnata showed the highest polyphenols, flavonoids and carotenoid content, with the highest antiradical activity.

Highly Efficient CRISPR/Cas9 Mediated Gene Editing in Ocimum basilicum 'FT Italiko' to Induce Resistance to Peronospora belbahrii.

Ocimum basilicum (sweet basil) is an economically important aromatic herb; in Italy, approximately 1000 ha of "Genovese-type" basil are grown annually in greenhouses and open fields and are subjected to Downy Mildew (DM) disease, caused by Peronospora belbahrii, leading to huge crop losses. Mutation of the Susceptibility (S) gene DMR6 (Downy Mildew Resistant 6) has been proven to confer a broad-spectrum resistance to DM. In this work, an effective Genome Editing (GE) approach mediated by CRISPR/Cas9 in O. basilicum 'Italiko', the élite cultivar used to produce "Pesto Genovese D.O.P", was developed. A highly efficient genetic transformation method mediated by A. tumefaciens has been optimized from cotyledonary nodes, obtaining 82.2% of regenerated shoots, 84.6% of which resulted in Cas9+ plants. Eleven T0 lines presented different type of mutations in ObDMR6; 60% were indel frameshift mutations with knock-out of ObDMR6 of 'FT Italiko'. Analysis of six T1 transgene-free seedlings revealed that the mutations of T0 plants were inherited and segregated. Based on infection trials conducted on T0 plants, clone 22B showed a very low percentage of disease incidence after 14 days post infection. The aromatic profile of all in vitro edited plants was also reported; all of them showed oxygenated monoterpenes as the major fraction.

Resilience of Stevia rebaudiana (Bertoni) Bertoni in the Underwater Biospheres of Nemo's Garden®: Adaptation to New Cultivation Systems.

The Nemo's Garden® project is an alternative production system for areas with scarce cultivable land but significant presence of water; thus, it is an interesting intervention to address the climate crisis. This work aimed to evaluate the micromorphological, biochemical, and phytochemical characteristics of Stevia rebaudiana (Bertoni) Bertoni grown underwater compared to the terrestrial specimens. The micromorphological analyses, performed on the leaves using light microscopy, fluorescence microscopy, and scanning electron microscopy, evidenced a general uniformity of the trichome morphotype and distribution pattern. The histochemical investigation indicated the simultaneous presence of terpenes and polyphenols in the trichome secreted material from the underwater samples and a prevailing polyphenolic content in the terrestrial specimens; this was also confirmed by biochemical analyses (26.6 mg GAE/g DW). The characterization of non-volatile components, performed using HPLC-MS, showed similar chemical profiles in all the samples, which were characterized by phenolic compounds and steviol glycosides. The volatile compounds, evaluated using HS-SPME coupled with GC-MS, showed sesquiterpene hydrocarbons as the main class in all the analyzed samples (80.1-93.9%). However, the control plants were characterized by a higher content of monoterpene hydrocarbons (12.1%). The underwater biosphere environment did not alter S. rebaudiana micro-morphological characters, although slight qualitative changes were evidenced for the compounds produced as a response to the growth conditions.

Postharvest Treatments on Sensorial and Biochemical Characteristics of Begonia cucullata Willd Edible Flowers.

Edible flowers (EFs) are currently consumed as fresh products, but their shelf life can be extended by a suitable drying technique, avoiding the loss of visual quality and valuable nutraceutical properties. Begonia cucullata Willd is a common ornamental bedding plant, and its leaves and flowers are edible. In this work, B. cucullata red flowers were freeze-dried (FD) and hot-air dried (HAD) at different temperatures. To the best of our knowledge, our study is the first one comparing different drying methodologies and different temperatures involving sensory characterization of EFs; therefore, a codified method for the description of the sensory profile of both fresh and dried B. cucullata was developed and validated. Phytochemical analyses highlighted the better preservation of antioxidant compounds (polyphenols, flavonoids and anthocyanins) for flowers dried at 60-70 °C. Visual quality was strongly affected by the drying treatments; in particular the color of the HAD samples significantly turned darker, whereas the FD samples exhibited a marked loss of pigmentation. Although all drying conditions led to a reduction in the hedonic indices if compared with fresh flowers, the best results in terms of organoleptic properties were obtained when the drying temperature was set to 60 or 70 °C.

Labdane Diterpenoids from Salvia tingitana Etl. Synergize with Clindamycin against Methicillin-Resistant Staphylococcus aureus.

Quorum-sensing (QS) is a regulatory mechanism in bacterial communication, important for pathogenesis control. The search for small molecules active as quorum-sensing inhibitors (QSI) that can synergize with antibiotics is considered a good strategy to counteract the problem of antibiotic resistance. Here the antimicrobial labdane diterpenoids sclareol (1) and manool (2) extracted from Salvia tingitana were considered as potential QSI against methicillin-resistant Staphylococcus aureus. Only sclareol showed synergistic activity with clindamycin. The quantification of these compounds by LC-MS analysis in the organs and in the calli of S. tingitana showed that sclareol is most abundant in the flower spikes and is produced by calli, while manool is the major labdane of the roots, and is abundant also in the leaves. Other metabolites of the roots were abietane diterpenoids, common in Salvia species, and pentacyclic triterpenoids, bearing a γ-lactone moiety, previously undescribed in Salvia. Docking simulations suggested that 1 and 2 bind to key residues, involved in direct interactions with DNA. They may prevent accessory gene regulator A (AgrA) binding to DNA or AgrA activation upon phosphorylation, to suppress virulence factor expression. The antimicrobial activity of these two compounds probably achieves preventing upregulation of the accessory gene regulator (agr)-regulated genes.

Abietane Diterpenoids from the Hairy Roots of Salvia corrugata.

Salvia corrugata Vahl. is an interesting source of abietane and abeo-abietane compounds that showed antibacterial, antitumor, and cytotoxic activities. The aim of the study was to obtain transformed roots of S. corrugata and to evaluate the production of terpenoids in comparison with in vivo root production. Hairy roots were initiated from leaf explants by infection with ATCC 15834 Agrobacterium rhizogenes onto hormone-free Murashige and Skoog (MS) solid medium. Transformation was confirmed by polymerase chain reaction analysis of rolC and virC1 genes. The biomass production was obtained in hormone-free liquid MS medium using Temporary Immersion System bioreactor RITA®. The chromatographic separation of the methanolic extract of the untransformed roots afforded horminone, ferruginol, 7-O-acetylhorminone and 7-O-methylhorminone. Agastol and ferruginol were isolated and quantified from the hairy roots. The amount of these metabolites indicated that the hairy roots of S. corrugata can be considered a source of these compounds.

Phytonutritional Content and Aroma Profile Changes During Postharvest Storage of Edible Flowers.

Edible flowers are niche horticultural products, routinely used as cooking ingredients in the food industry. Currently, new species are required with the aim of enlarging the number of species with a long shelf-life, healthy nutraceutical compounds, and new fragrance and tastes. Ageratum houstonianum Mill, Tagetes lemmonii A. Gray, Salvia dorisiana Standl, and Pelargonium odoratissimum (L.) L'Hér "Lemon" were selected for their different morphological characteristics and color. Fresh flowers were analyzed to characterize their phytonutritional content and aroma profile. Postharvest was determined up to 6 days of cold storage at 4°C in transparent polypropylene boxes. Visual quality and cellular membrane damage were observed. The relative content of different antioxidant constituents (e.g., polyphenols, flavonoids, anthocyanins, ascorbic acid), nutritional compounds (soluble sugars, crude proteins), the antioxidant scavenging activity, and the volatile profile were determined and correlated to the quality of shelf-life of the different species. The yellow T. lemmonii freshly picked flowers showed the highest ascorbic acid and flavonoids content, which was maintained during the cold storage, as well as the best visual quality. Limited changes in metabolites were detected in the light blue A. houstonianum during postharvest, although the visual quality is severely compromised. Magenta S. dorisiana and light pink P. odoratissimum showed similar changes in antioxidant constituents during cold storage. For the first time, the volatile compounds have been identified in the four species. Sesquiterpene hydrocarbons are the main class in fresh flowers of A. houstonianum, S. dorisiana, and P. odoratissimum, while monoterpene hydrocarbons are abundant in T. lemmonii. The cold storage influenced mainly P. odoratissimum and S. dorisiana flavor initially dominated by the increase in total monoterpenes at 6 days, reaching a relative content of 90%. Both A. houstonianum and T. lemmonii conserved the prevalence of the same class of constituents in all the analyzed conditions, even though the cold storage influenced the major compound abundance. On the basis of the results, T. lemmonii was the most interesting species with the longest shelf-life due to its phytonutritional and aromatic constituents. Results indicated the peculiar metabolic and physiological attitude of flowers species to cold storage.

Bioactive Compounds and Aroma Profile of Some Lamiaceae Edible Flowers.

Edible flowers are consumed for their appearance, colours, nutritional and healthy properties, but the use is limited by the actual number of the species. Seven edible flowers of the Lamiaceae family (Ocimeae and Mentheae tribes) were investigated: Monarda didyma 'Fireball', Nepeta × faassenii 'Six Hills Giant', Ocimum basilicum 'Blue Spice', O. basilicum 'Cinnamon', Ocimum × citriodorum, Salvia discolor, and Salvia microphylla 'Hot Lips'. Total soluble sugars, proteins, polyphenols, carotenoids, ascorbic acid and antioxidant activity were detected. The species of the Mentheae tribe contained higher sugar content than Ocimeae flowers, the opposite with regard to protein content. Ocimeae tribe flowers showed high polyphenols and carotenoids content. The Ocimeae tribe together with two specie of the Mentheae tribe showed an aroma profile dominated by sesquiterpene hydrocarbons (58.0% in S. discolor to 77.9% in Ocimum × citriodorum). Oxygenated monoterpenes prevailed in Nepeta and Monarda, also present in the essential oil of this latter species (84.5%). By contrast, Nepeta and S. discolor evidenced non-terpenes as the principal class (41.2% and 77.5%, respectively), while the oxygenated sesquiterpene was the main one in S. microphylla. The two varieties of Ocimum spp. showed oxygenated monoterpenes as the main class of volatiles.

Trace Elements in Edible Flowers from Italy: Further Insights into Health Benefits and Risks to Consumers.

The use of edible flowers in cooking dates back to ancient times, but recently it is gaining success among the consumers, increasingly attentive to healthy and sustainable foods of high quality, without neglecting taste, flavour, and visual appeal. The present study aims to deepen the knowledge regarding the mineral composition of edible flowers, an aspect not widely investigated in scientific literature. The concentrations of Cd, Co, Cu, Fe, Mn, Ni, Pb, Sr, V, and Zn have been determined by Inductively Coupled Plasma Optical Emission Spectrometry (ICP OES) in flowers belonging to a wide variety of species. The study highlights that some floral species are characterized by significantly higher concentrations of certain trace elements, e.g., the flowers of Acmella oleracea for Mn, those of basil (Ocimum basilicum) and of pumpkins (Cucurbita moschata and C. pepo) for Cu and Sr, and those of orange daylily (Hemerocallis fulva) for Ni. Potentially toxic elements are present at low concentrations, often below the limit of the detection for Cd, Co, Ni, V. In all samples, Cd and Pb are well below the maximum permitted levels in foodstuffs. It can be concluded that the edible flowers analyzed can be considered a good source of essential elements and do not present risks for the consumer health as for the mineral composition.

Volatilomic Analysis of Four Edible Flowers from Agastache Genus.

Volatilomes emitted from edible flowers of two species of Agastache (A. aurantiaca (A.Gray) Lint & Epling, and A. mexicana (Kunth) Lint & Epling) and from two hybrids (Agastache 'Arcado Pink' and Agastache 'Blue Boa') were investigated using a solid-phase microextraction technique as well as the extraction of its essential oils. Oxygenated monoterpenes were almost always the predominant class (>85%) of volatile organic compounds (VOCs) in each sample of A. aurantiaca, A. 'Blue Boa' and A. mexicana, with the exception of A. 'Arcado Pink' (38.6%). Pulegone was the main compound in A. aurantiaca (76.7%) and A. 'Blue Boa' (82.4%), while geranyl acetate (37.5%) followed by geraniol (16%) and geranial (17%) were the principal ones in A. mexicana. The essential oil composition showed the same behavior as the VOCs both for the main class as well as the major constituent (pulegone) with the same exception for A. mexicana. Total soluble sugars, secondary metabolites (polyphenols, flavonoids and anthocyanins) and antioxidant activity were also investigated to emphasize the nutraceutical properties of these edible flowers.

Essential Oil Composition and Volatile Profile of Seven Helichrysum Species Grown in Italy.

Helichrysum genus consists of about 600 species widespread throughout the world, especially in South Africa and in the Mediterranean area. In this study the aroma profile (HS-SPME) and the EO compositions of seven Helichrysum species (H. cymosum, H. odoratissimum, H. petiolare, H. fontanesii, H. saxatile, H. sanguineum, and H. tenax) were evaluated. All the plants were grown in Italy under the same growth conditions. The volatile constituents, particularly monoterpenes, depended by the plant's genotype and ecological adaptation. This study represents the first headspace evaluation on the selected plants and the results evidenced that monoterpenes represented the main class of constituents in five of the seven species analysed (from 59.2% to 95.0%). The higher content in sesquiterpene hydrocarbons was observed in the Mediterranean species of H. sanguineum (68.0%). Only H. saxatile showed relative similar abundance of monoterpenes and sesquiterpene hydrocarbons. The essential oil composition of the majority of examined species are characterised by high percentage of sesquiterpenes (especially ß-caryophyllene and δ-cadinene) ranging from 51.3% to 92.0%, except for H. cymosum, H. tenax, and H. sanguineum leaves where monoterpenes predominated (from 51.7% to 74.7%).

Activity of Salvia dolomitica and Salvia somalensis Essential Oils against Bacteria, Molds and Yeasts.

Essential oils (EOs) from Salvia dolomitica and Salvia somalensis, widely employed in the cosmetic and perfume industry, were analyzed for composition and tested against bacterial and fungal pathogens isolated from clinical and environmental specimens. The analyses were carried out against Staphylococcus aureus, Staphylococcus pseudointermedius, Pseudomonas aeruginosa, Escherichia coli, Streptococcus canis, Streptococcus pyogenes, Klebsiella pneumoniae, Proteus mirabilis, Microsporum canis, Microsporum gypseum, Trichophyton mentagrophytes, Aspergillus niger, Aspergillus flavus, Candida albicans, Candida krusei, Mucor sp. and Trichothecium roseum. Both EOs showed similar percentages of total monoterpenes and sesquiterpene hydrocarbons. The main constituents were 1,8-cineole and ß-caryophyllene in S. dolomitica and bornyl acetate and camphor in S. somalensis. The selected EOs have no relevant antifungal or antibacterial activities if compared to conventional drugs.

Micropropagation of Salvia wagneriana Polak and hairy root cultures with rosmarinic acid production.

Salvia wagneriana Polak is a tropical species native to Central America, well adapted to grow in the Mediterranean basin for garden decoration. Micropropagation has been assessed from axillary shoots of adult plants using a Murashige and Skoog basal medium, with the addition of 1.33-µM 6-benzylaminopurine for shoot proliferation; the subsequent rooting phase occurred in plant growth regulator-free medium. The plants were successfully acclimatised with high survival frequency. Hairy roots were induced after co-cultivation of leaf lamina and petiole fragments with Agrobacterium rhizogenes and confirmed by PCR. The establishment and proliferation of the selected HRD3 line were obtained in hormone-free liquid medium and the production of rosmarinic acid (RA) was evaluated after elicitation. The analysis of RA was performed by LC-ESI-DAD-MS in the hydroalcoholic extracts. The addition of casein hydrolysate increased the RA production, whereas no enrichment was observed after the elicitation with jasmonic acid.

Establishment and analysis of in vitro biomass from Salvia corrugata Vahl. and evaluation of antimicrobial activity.

Demethylfruticuline A and fruticuline A, the most abundant compounds from the surface extract of Salvia corrugata Vahl., have shown antibacterial, antitumor and cytotoxic activities. In order to obtain these icetexane diterpenes from in vitro cultures of S. corrugata, protocols were developed for callus production, micropropagation and shoot regeneration. Analysis of the regenerated shoots showed the presence of both icetexanes, micropropagated plants contained only fruticuline A, while the callus contained trace amounts of both diterpenes. The yield of fruticuline A was higher in the methanolic extract of regenerated shoots than in those of fresh leaves and fresh shoot tips. In addition to these diterpenes, the regenerated shoot and micropropagated plant extracts afforded seven other diterpenes, one icetexane and six abietanes, identified by UV, IR, 1D- and 2D-NMR and HR-MS analysis. Five compounds (19-acetoxy-7α-hydroxyroyleanone, 7ß,20-epoxy-11,12,19-trihydroxyabieta-8,11,13-triene, 7,20-dihydrofruticuline A, 7ß-acetoxy-20-hydroxy-19,20-epoxyroyleanone, 7ß-ethoxy-6ß,20:19,20-diepoxyroyleanone) were previously undescribed. Although the crude plant surface extract did not possess any antibacterial activity, methanolic extracts of in vitro tissues and two compounds, namely 7ß-acetoxy-20-hydroxy-19,20-epoxyroyleanone and 7ß-ethoxy-6ß,20:19,20-diepoxyroyleanone, isolated in suitable amounts, were active in varying degrees against multidrug resistant clinical strains of Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis and Enterococcus faecium, displaying MIC values ranging from 32, 64 to 128µg/mL.

Somatic Embryogenesis in Lisianthus (Eustoma russellianum Griseb.).

Somatic embryogenesis is, for the main floricultural crops, a promising system for commercial scale-up, providing cloned material to be traded as seedlings. Somatic embryos, having the contemporary presence of root apical meristem and shoot apical meristem, can be readily acclimatized. For Lisianthus it is possible to induce embryogenic callus from leaf fragments of selected genotypes and to obtain embryos either in agarized substrate or in liquid suspension culture. The production of somatic embryos in liquid medium is high and can be modulated in order to synchronize the cycle and the size of the neoformed structures. The possibility to use the liquid substrate with high propagation rates reduces labor costs and could support the costs of eventual automation. In this paper we report a stepwise protocol for somatic embryogenesis in the species Eustoma russellianum.

In vitro cultures of Bituminaria bituminosa: pterocarpan, furanocoumarin and isoflavone production and cytotoxic activity evaluation.

Bituminaria bituminosa L. is known for producing several compounds with considerable pharmaceutical interest, such as phenylpropanoids, furanocoumarins and pterocarpans. In vitro cultures of seedlings, shoots, and callus have been produced to obtain plant materials useful for the production of these metabolites. The secondary metabolite profile was evaluated by HPLC-DAD. The extracts of all the in vitro material contained the flavonoid daidzein, while plicatin B, erybraedin C and bitucarpin A were found only in the extracts of the in vitro shoots and in wild shoots. The furanocoumarins angelicin and psoralen were found in in vivo and in vitro plants, but in the callus were not detectable. The extracts were also tested for cytotoxic activity in HeLa cell culture; the highest level of cytotoxicity was found in in vitro shoot extracts.

In vitro propagation of Hydrangea spp.

Hydrangea (Hortensia) is a highly popular ornamental plant for garden decoration, and now it is commercially produced for cut flower branches. For in vitro culture, Murashige and Skoog medium supplemented with BA (0.25 mg/L) and sucrose (30 g/L) was used. Culture conditions were 23 ± 1°C of temperature, light intensity of 35 µmol/m(2)/s P.P.F.D., and 16/8 h day/night photoperiod. Following shoot proliferation, the in vitro rooting frequency was 100% on a medium containing NAA 0.5 mg/L. However, 95% direct in vivo rooting was achieved by dipping microcuttings in a 5,000 ppm K-IBA solution which were transferred afterward to a glasshouse for acclimatization. After 21 days, fully acclimatized and well-established plants were obtained, suitable for commercialization. Furthermore, leaf fragments derived from in vitro plantlets were cultured for callus induction and adventitious shoot regeneration.

Production of Curcuminoids in different in vitro organs of Curcuma longa.

Curcuma longa L. (turmeric) is one of the most important spice and safe food additives. Its main constituents, curcuminoids, showed anti-inflammatory, antitumor and antioxidant effects. In the present work, an in vitro propagation method was developed to achieve selected plant organs with quantified curcuminoid content. In vitro plants were obtained from sprouting buds as primary explants. The major curcuminoid constituents, such as curcumin (CUR), demethoxycurcumin (DEM), and bis-demethoxycurcumin (bis-DEM) were examined in different organs by LC-DAD-ESI-MS. A significant production of curcumin (more than 260 microg g(-1) fresh weight) was obtained from in vitro microrhizomes, especially grown in a Murashige and Skoog medium (MS) supplemented with kinetin (0.1 mg L((-1)), alpha-naphthaleneacetic acid (NAA, 1 mg L(-1)), sucrose (6%), agar (5%) and activated charcoal (0.1%). The analyzed microrhizomes showed reduced amounts of DEM and bis-DEM in comparison with CUR levels. In addition a shoot culture line was suitable to biosynthesize curcuminoids, in a ratio very similar to that identified in the fresh rhizomes of parent plants. This study represents the first direct quantification of curcuminoids in turmeric in vitro shoots and microrhizomes to be used in dietary supplements.

In vitro propagation of ornamental myrtus (Myrtus communis).

The Myrtle (Myrtus communis L.) is an evergreen shrub typical of the Mediterranean area; it is an interesting plant with multipurpose use. The ornamental use takes into account the production of green cut branches for indoor decoration and production of pot plants for gardening. In this species, there is a great variability in the natural germplasm around the Mediterranean coasts for type and size of fruit, plant architecture, leaf size and internode length. Selected genotypes have been successfully sterilized and cultured in vitro. The shoots were multiplied on MS (16) salts and vitamins, with 0.5 mg/L BA and 0.2 mg/L IAA. Clones showed variation of multiplication rate and rooting percentage. IAA or IBA at 0.5 mg/L increased the rooting percentage and noticed differences in root number and length. The sucrose concentration can affect rooting, such as light intensity during the in vitro rooting phase can modulate biomass production and chlorophyll content. The combination of these factors enhanced the frequency rate of acclimatization.