The Dark Side of the pollen: BSA-seq identified genomic regions linked to male sterility in globe artichoke.

Globe artichoke (Cynara cardunculus var. scolymus; 2n = 2x = 34) is a food crop consumed for its immature flower heads. Traditionally, globe artichoke varietal types are vegetatively propagated. However, seed propagation makes it possible to treat the crop as annual, increasing field uniformity and reducing farmers costs, as well as pathogens diffusion. Despite globe artichoke's significant agricultural value and the critical role of heterosis in the development of superior varieties, the production of hybrids remains challenging without a reliable system for large-scale industrial seed production. Male sterility (MS) presents a promising avenue for overcoming these challenges by simplifying the hybridization process and enabling cost-effective seed production. However, within the Cynara genus, genic male sterility has been linked to three recessive loci in globe artichoke, with no definitive genetic mechanism elucidated to date. A 250 offsprings F2 population, derived from a cross between a MS globe artichoke and a male fertile (MF) cultivated cardoon (C. cardunculus var. altilis) and fitting a monogenic segregation model (3:1), was analyzed through BSA-seq, aiming at the identification of genomic regions/genes affecting male sterility. Four QTL regions were identified on chromosomes 4, 12, and 14. By analyzing the sequence around the highest pick on chromosome 14, a cytochrome P450 (CYP703A2) was identified, carrying a deleterious substitution (R/Q) fixed in the male sterile parent. A single dCAPS marker was developed around this SNP, allowing the discrimination between MS and MF genotypes within the population, suitable for applications in plant breeding programs. A 3D model of the protein was generated by homology modeling, revealing that the mutated amino acid is part of a highly conserved motif crucial for protein folding.

Byproducts of Globe Artichoke and Cauliflower Production as a New Source of Bioactive Compounds in the Green Economy Perspective: An NMR Study.

The recovery of bioactive compounds from crop byproducts leads to a new perspective way of waste reutilization as a part of the circular economy. The present study aimed at an exhaustive metabolite profile characterization of globe artichoke and cauliflower byproducts (leaves, stalks, and florets for cauliflower only) as a prerequisite for their valorization and future implementations. The metabolite profile of aqueous and organic extracts of byproducts was analyzed using the NMR-based metabolomics approach. Free amino acids, organic acids, sugars, polyols, polyphenols, amines, glucosinolates, fatty acids, phospho- and galactolipids, sterols, and sesquiterpene lactones were identified and quantified. In particular, globe artichoke byproducts are a source of health-beneficial compounds including chiro-inositol (up to 10.1 mg/g), scyllo-inositol (up to 1.8 mg/g), sesquiterpene lactones (cynaropicrin, grosheimin, dehydrocynaropicrin, up to 45.5 mg/g in total), inulins, and chlorogenic acid (up to 7.5 mg/g), whereas cauliflower byproducts enclose bioactive sulfur-containing compounds S-methyl-L-cysteine S-oxide (methiin, up to 20.7 mg/g) and glucosinolates. A variable content of all metabolites was observed depending on the crop type (globe artichoke vs. cauliflower) and the plant part (leaves vs. stalks). The results here reported can be potentially used in different ways, including the formulation of new plant biostimulants and food supplements.

Polyphenolic Compound Variation in Globe Artichoke Cultivars as Affected by Fertilization and Biostimulants Application.

Globe artichoke is an ancient herbaceous plant native to the Mediterranean Basin. The edible part of the plant is limited to the fleshy leaves (bracts) and receptacle of a large immature inflorescence (head) that has been shown to be a rich source of bioactive compounds. Nutritional and pharmacological properties of artichoke heads and leaves are attributed mainly to polyphenolic compounds and inulin present at high concentration. In this study, polyphenols were investigated in two artichoke cultivars (Opal and Madrigal) in response to four nitrogen rates and foliar applications of biostimulating products under drip irrigation. Field experiments were carried out over two growing seasons (2015-2016, 2016-2017) in Policoro (MT), Southern Italy, on a deep clay soil in sub-humid climate conditions. Phenolic compounds were isolated and characterized by means of high-performance liquid chromatography with photodiode array detection and electrospray ionization/mass spectrometry (HPLC-DAD-MS/MS) analysis. In both cultivars, caffeoylquinic acids were more abundant when a dose of 100 kg ha-1 of ammonium nitrate was provided, whereas apigenins were not affected by nitrogen fertilization. Luteolins increased in cv Opal and decreased in cv Madrigal following N fertilization. The application of biostimulants (3 L ha-1) favored the accumulation of polyphenols, in particular of caffeoylquinic acids and apigenin, in artichoke heads in both cultivars. The results obtained highlight some positive aspects related to the synergistic effect of nitrogen fertilization and biostimulant foliar application.

In vivo study of the bioavailability and metabolic profile of (poly)phenols after sous-vide artichoke consumption.

Artichokes are a rich source of (poly)phenols, mainly caffeoylquinic acids, but little is known about their bioavailability from this source. This study investigated the absorption, metabolism and excretion of (poly)phenols after sous-vide artichoke consumption (5776 µmol of (poly)phenols) by healthy volunteers. Seventy-six (poly)phenol metabolites were identified by UHPLC-MS/MS using authentic standards, including acyl-quinic acids plus C6-C3, C6-C1, C6-C2, C6-C1-N, C6-C0 metabolites, and their phase-II conjugates. The major metabolites were 3'-methoxy-4'-hydroxycinnamic acid, 3'-methoxycinnamic acid-4'-sulfate, and 4'-hydroxycinnamic acid-3'-sulfate, which appeared early in plasma (Tmax < 4 h); plus 3-(3'-methoxy-4'-hydroxyphenyl)propanoic acid, 3-(4'-methoxyphenyl)propanoic acid-3'-glucuronide, 3-(3'-hydroxyphenyl) propanoic acid and hippuric acids, which appeared later (Tmax > 6 h). The 24 h urinary recovery averaged 8.9% (molar basis) of the (poly)phenols consumed. Hepatic beta-oxidation of 3',4'-dihydroxycinnamic acid and methylated conjugates occurred, but was limited (<0.04%). 3'-Methylation exceeded 4'-methylation and interindividual variability was high, especially for gut microbial metabolites (up to 168-fold).

Botanical authentication of globe artichoke-containing foods: Differentiation of Cynara scolymus by a novel HRM approach.

Cynara scolymus L., known as globe artichoke, is a medicinal plant widely used in plant food supplements (PFS) and herbal infusions due to its beneficial health properties. The high demand for artichoke-containing products can lead to adulteration practices. In this work, a real-time polymerase chain reaction (PCR) system coupled to high-resolution melting (HRM) analysis was proposed to differentiate C. scolymus from other Cynara species. Hence, a Cynara-specific real-time PCR assay was successfully developed with high analytical performance, achieving a sensitivity of 0.4 pg of globe artichoke DNA. HRM analysis enabled the discrimination of C. scolymus, with a high level of confidence (>98%), corroborating sequencing data. Application results to artichoke-containing PFS and mixed herbal infusions allowed confirming the presence of C. scolymus in 38% of the samples, suggesting the substitution/mislabelling of globe artichoke in 2 samples and the need for further efforts to increase DNA amplifiability of PFS.

Active Packaging-Releasing System with Foeniculum vulgare Essential Oil for the Quality Preservation of Ready-to-Cook (RTC) Globe Artichoke Slices.

Two globe artichoke genotypes, "Spinoso sardo" and "Opera F1", have been processed as ready-to-cook (RTC) slices and refrigerated at 4 °C for 12 days (i) to evaluate the suitability to be processed as RTC slices; (ii) to evaluate the effect of a Foeniculum vulgare essential oil (EO) emitter, within an active package system, to delay quality decay, thus extending shelf life; (iii) to estimate the impact of EO emitter on the sensory profile of the RTC slices after cooking. Results revealed that both globe artichoke genotypes possess a good attitude to be processed as RTC product. "Opera F1" showed the best performances for color parameters, texture and chemical indexes, while "Spinoso sardo" showed lower mass loss (ML) over the storage time. The addition of EO emitter slowed down the consumption of O2, better preserved texture when compared to the control and more effectively control polyphenol oxidase (PPO) activity and antioxidants' retention during the cold storage. Microbial counts in control globe artichoke RTC slices were significantly higher than those packed with EO emitter, confirming the inhibiting role played by EO of F. vulgare. In addition, the EO emitter did not influence negatively the sensory profile of RTC globe artichoke slices after microwave cooking.

Artichoke Biorefinery: From Food to Advanced Technological Applications.

A sequential extraction process has been designed for valorizing globe artichoke plant residues and waste (heads, leaves, stalks, and roots left in the field) by means of green extraction techniques according to a biorefinery approach. We investigated two cascading extractions based on microwave-assisted extractions (MAE) and green solvents (water and ethanol) that have been optimized for varying temperature, solvent and extraction time. In the first step, phenols were extracted with yields that ranged between 6.94 mg g-1 dw (in leaves) and 3.28 mg g-1 dw (in roots), and a phenols productivity of 175.74 Kg Ha-1. In the second step, inulin was extracted with impressive yields (42% dw), higher than other conventional inulin sources, corresponding to an inulin productivity of 4883.58 Kg Ha-1. The remaining residues were found to be valuable feedstocks both for bioenergy production and green manure (back to the field), closing the loop according to the Circular Economy paradigm.

Silage Fermentation: A Potential Biological Approach for the Long-Term Preservation and Recycling of Polyphenols and Terpenes in Globe Artichoke (Cynara scolymus L.) By-Products.

An economic and effective method for storage is necessary to make full use of the nature of active components in artichoke by-products and ease environmental pressure. In this paper, the potential of silage fermentation for the preservation and recycling of polyphenols and terpenes in artichoke by-products is evaluated. The silage of artichoke by-products is characterized by lactic acid bacteria fermentation. Silage distinctly increases the abundance of lactic acid bacteria in artichoke by-products, such as Lactobacillus, Lactococcus, Serratia, and Weissella, and greatly increases the abundance of Firmicutes. The improvement of the microorgan structure and composition is of great significance for the quality of artichoke by-products. Polyphenols in the stems and leaves of artichokes are preserved well in silage. Among the 18 polyphenol compounds detected by high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (HPLC-QqQ-MS/MS), the contents of 11 phenolic acids and four flavonoids increased significantly. For terpenes detected by gas chromatography-mass spectrometry (GC-MS), the contents of four pentacyclic triterpenoids increased significantly, while two sterols were kept stable in the silage process. Silage is a potential biotechnology for the long-term preservation of bioactive components, such as polyphenols and terpenes in artichoke by-products, and the results provide a scientific basis for the efficient utilization of by-products.

Cryopreservation of shoot apices and callus cultures of globe artichoke using vitrification method.

BACKGROUND: Cryogenic cooling became a crucial tool for the storage of heterozygous plants such as globe artichoke. This study was carried out to optimize a reliable method for in vitro cryopreservation of shoot apices and callus cultures of globe artichoke using dimethylsulfoxide (DMSO) and Plant Vitrification Solutions 2 (PVS2) as cryoprotectant solutions. Shoot apices were exposed to DMSO or PVS2 for 20, 40, 60, and 80 min prior to plunge in liquid nitrogen (LN). RESULTS: It was found that using PVS2 as a cryoprotectant in cryopreservation of shoot apices of globe artichoke was more effective compared with using of DMSO alone. Among the exposure time tested, 60 min gave the best results of survival. The highest survival (60%), regeneration (56%), and proliferated shootlets (4.30) were obtained after cryoprotection with PVS2 for 60 min. Regarding callus cultures, the maximum values of fresh weights and subsequently growth value of recovered callus were registered with 40 min followed by 60 min exposure time. Related to the type of the tested cryoprotectants, the best survival and growth parameters of the cryopreserved callus cultures were obtained with PVS2 treatments. Treatment with PVS2 for 40 min registered the highest survival observations of cryopreserved callus. Also, the maximum values of fresh weight (1.30 g) and growth value (4.20) were obtained with 40 min exposure time. Microscopy analysis presented as cell morphology revealed that the treatment of PVS2 40% was the optimum for cell growth of cryopreserved callus of globe artichoke. CONCLUSION: The results demonstrated that using PVS2 as a cryoprotectant in cryopreservation of shoot apices and callus cultures of globe artichoke was more effective compared with DMSO.

Shelf-life study of ready-to-cook slices of globe artichoke 'Spinoso sardo': effects of anti-browning solutions and edible coating enriched with Foeniculum vulgare essential oil.

BACKGROUND: The globe artichoke [Cynara cardunculus var. scolymus (L.) Fiori] is a natural source of minerals, fiber, inulin, and polyphenols. A ready-to-cook (RTC) version of this product could improve its commercialization and consumption. In this study, the effects of (i) anti-browning treatments (citric acid 0.5% + ascorbic acid 2%, or cysteine 0.5%, w/v) and (ii) dipping in locust bean gum (LBG) edible coating with or without Foeniculum vulgare essential oil (EO) were evaluated on RTC globe artichoke slices cv. 'Spinoso sardo' during storage (11 days at 4 °C). Fresh weight loss, color, texture, polyphenol oxidase (PPO), microbiological and chemical parameters, antioxidant capacity, and sensory descriptors were determined. RESULTS: The results showed that, despite the antioxidant treatment used, when EO was added to LBG, all microbiological groups underwent an average reduction of 0.50 log CFU g-1 . Cysteine preserved the antioxidant capacity, color, and texture of samples better than citric acid and ascorbic acid, and inhibited PPO; the addition of EO to LBG enhanced these effects. At the end of the storage period, the overall highest sensory attribute score was attained by the coated globe artichoke samples with EO. CONCLUSION: The marked efficacy of the LBG edible coating with added F. vulgare EO in maintaining microbiological, physical, chemical, and sensory qualities makes it a promising processing tool for the preservation of ready-to-cook Spinoso sardo globe artichoke slices during storage for 11 days at 4 °C. © 2019 Society of Chemical Industry.

Bioactivities, chemical composition and nutritional value of Cynara cardunculus L. seeds.

In the present study, the nutritional value, bioactive properties, and chemical composition of various cardoon (Cynara cardunculus L.) genotypes cultivated in central Greece were investigated. The results demonstrated that Cynara seeds are a good source of fat and protein, while they also contain considerable amounts of K, Mg, and Fe and low amount of Na. Sucrose, oxalic acid, and α-tocopherol were the only free sugar, organic acid, and tocopherol isoform respectively, found among the studied genotypes. The most abundant fatty acids were linoleic, oleic and palmitic acid, while PUFA was the most abundant fatty acid class. All the tested seeds contained only two phenolic compounds, namely 5-O-caffeoylquinic acid and 3,5-O-caffeoylquinic acid, while significant antioxidant activities and cytotoxicity against tumor cell lines and antimicrobial effects were also observed. In conclusion, cardoon seed extracts could be exploited in the food and pharmaceutical industries as alternative sources of natural compounds with bioactive properties.

Analysis of DNA Methylation Patterns Associated with In Vitro Propagated Globe Artichoke Plants Using an EpiRADseq-Based Approach.

Globe artichoke represents one of the main horticultural species of the Mediterranean basin, and 'Spinoso sardo' is the most widespread and economically relevant varietal type in Sardinia, Italy. In the last decades, in vitro culture of meristematic apices has increased the frequency of aberrant plants in open-field production. These off-type phenotypes showed highly pinnate-parted leaves and late inflorescence budding, and emerged from some branches of the true-to-type 'Spinoso sardo' plants. This phenomenon cannot be foreseen and is reversible through generations, suggesting the occurrence of epigenetic alterations. Here, we report an exploratory study on DNA methylation patterns in off-type/true-to-type globe artichoke plants, using a modified EpiRADseq technology, which allowed the identification of 2,897 differentially methylated loci (DML): 1,998 in CG, 458 in CHH, and 441 in CHG methylation contexts of which 720, 88, and 152, respectively, were in coding regions. Most of them appeared involved in primary metabolic processes, mostly linked to photosynthesis, regulation of flower development, and regulation of reproductive processes, coherently with the observed phenotype. Differences in the methylation status of some candidate genes were integrated with transcriptional analysis to test whether these two regulation levels might interplay in the emergence and spread of the 'Spinoso sardo' non-conventional phenotype.

Modification of the existing maximum residue levels for mandipropamid in various crops.

In accordance with Article 6 of Regulation (EC) No 396/2005, the applicants Syngenta Crop Protection B.V. and Agriculture and Horticulture Development Board (AHDB) submitted, respectively, a request to the competent national authorities in the Netherlands and United Kingdom to modify the existing maximum residue levels (MRLs) for the active substance mandipropamid in various crops. The data submitted in support of the request were found to be sufficient to derive MRL proposals for beetroots, radishes, cauliflowers, Brussels sprouts, witloofs/Belgian endives, peas (without pods) and globe artichokes. Adequate analytical methods for enforcement are available to control the residues of mandipropamid in plant matrices under consideration at the validated limit of quantification (LOQ) of 0.01 mg/kg. Based on the risk assessment results, EFSA concluded that the intake of residues resulting from the use of mandipropamid according to the reported agricultural practices is unlikely to present a risk to consumer health.

Nutritional value and chemical composition of Greek artichoke genotypes.

The nutritional value and chemical composition of various artichoke genotypes cultivated in Greece were evaluated. Significant differences were detected in water content, as well as in fat, ash, protein and carbohydrate content. Proteins and carbohydrates were detected in high amounts in all the genotypes. Antioxidant activity was also varied between the studied samples. Palmitic and linoleic acids were the most abundant fatty acids in artichoke heads, while stearic, oleic, alpha-linolenic, arachidic, behenic and lignoceric acids were detected in lesser amounts. 3,5-O-Dicaffeoylquinic acid and 5-O-caffeoylquinic acid were the main phenolic compounds. In conclusion, heads of artichoke genotypes cultivated in Greece showed a high nutritional value and antioxidant activity which signifies the importance of this vegetable for the Mediterranean diet, while the diversity in chemical composition between the studied samples should be further exploited for the selection of elite cultivars with specific end-uses of the final product.

Artichoke edible parts are hepatoprotective as commercial leaf preparation

ABSTRACT Chemical profile analyses of artichoke (Cynara scolymus L., Asteraceae) edible parts (fleshy receptacle, inner bracts) as well as roots are compared with the commercially usable leaf extract using HPLC-DAD-ESI-MS via chlorogenicacid as a marker. Overall polyphenolic constituents demonstrated by means of LC/MS profiling. The nutritional values and inulin contents of different assessed parts were investigated. The present study was designed to determine the effect of artichoke: leaves, bracts, receptacles and roots alcoholic extracts against CCl4-induced acute hepatotoxicity and hyperlipidemia in rats by means of histopathological and biochemical parameters. Serum liver enzymes levels of aspartate amino transferase, alanine amino transferase, alkaline phosphatase and lipid peroxidase content (malondialdehyde MDA) were estimated. Blood glutathione, total cholesterol, triacylglycerides and high density lipid level were estimated in plasma. The ethanol extract of roots, leaves, bracts and receptacles were standardized to (0.82 ± 0.02, 1.6 ± 0.06, 2.02 ± 0.16 and 2.4 ± 0.27 mg chlorogenic acid/100 mg extract), respectively. The receptacle showed the highest content of polyphenols and exhibits the highest antioxidant activity. HPLC analysis of inulin in the receptacles of globe artichoke revealed high content of inulin (41.47 mg/g) dry extract. All artichoke parts contain comparable vitamins and minerals. Artichokes receptacles extract when taken in dose of (500 mg/kg/day) reduce the lesion caused by CCl4 alone more than groups receiving silymarin. Bracts and leaves extract exert nearly the same effect.

Effects of alternative cropping systems on globe artichoke qualitative traits.

BACKGROUND: Traditionally, globe artichoke cultivation in the Mediterranean basin is based on monoculture and on use of high amounts of nitrogen fertiliser. This raises issues regarding its compatibility with sustainable agriculture. We studied the effect of one typical conventional (CONV) and two alternative cropping systems [globe artichoke in sequence with French bean (NCV1), or in biannual rotation (NCV2) with cauliflower and with a leguminous cover crop in inter-row spaces] on yield, polyphenol and mineral content of globe artichoke heads over two consecutive growing seasons. RESULTS: NCV2 showed statistical differences in terms of fresh product yield with respect to the monoculture systems. In addition, the dihydroxycinnamic acids and dicaffeoylquinic acids of non-conventional samples were one-fold significantly higher than the conventional one. All the samples reported good mineral content, although NCV2 achieved a higher Fe content than conventional throughout the two seasons. After two and three dates of sampling, the CONV samples showed the highest levels of K content. CONCLUSION: In our study, an acceptable commercial yield and quality of 'Spinoso sardo' were achieved by shifting the common conventional agronomic management to more sustainable ones, by means of an accurate choice of cover crop species and rotations introduced in the systems. © 2017 Society of Chemical Industry.

Minerals profile of two globe artichoke cultivars as affected by NPK fertilizer regimes.

Globe artichoke is a proven source of various minerals (such as K, Fe and Zn) in the Mediterranean diet, but their content in response to fertilizer regime has not yet been investigated sufficiently. Thus, we monitored the effect of two contrasting nitrogen/phosphorus/potassium (NPK) fertilizer regimes (one balanced and the other excessive) on the minerals accumulation of 'Apollo' and 'Tema 2000' cultivars, grown in three Sicilian locations ('Landolina', 'Iannarello' and 'Zotto') - South Italy. Except for total nitrogen, the balanced fertilizer regime favoured the accumulation of both macro- and micro-minerals, but with a different extent depending especially on trial location. Particularly, plants grown at 'Iannarello' responded more strongly to the fertilizer regime with respect to K, P, Ca, Fe and Zn accumulation, as a result of its different soil characteristics than the other locations. Providing a balanced supply of nitrogen/phosphorus/potassium via fertilization can enhance the nutritive value of globe artichoke, but taking into account especially soil characteristics.

Determination of pesticide residues in globe artichoke leaves and fruits by GC-MS and LC-MS/MS using the same QuEChERS procedure.

Aiming to select the most suitable sample preparation for the multiresidue analysis of pesticides in globe artichoke (Cynara cardunculus L.), a modified QuEChERS, a matrix solid phase dispersion and a dispersive ethyl acetate extraction were compared. Trueness and precision were determined at 0.2mgkg-1 for the three methods. The modified QuEChERS showed the best performance. The scope of the method was enlarged to 35 GC and 63 LC amenable pesticides, its overall performance was evaluated and validated to artichoke leaves and fruits according to DG-SANTE Guidelines. Different matrix effects were observed for most of the pesticides which were higher for leaves than fruits. Difenoconazole and flutriafol suffered signal suppression in leaves extracts but showed positive matrix effects in fruits. All pesticides were analyzed at or below their Maximum Residue Levels fixed for globe artichoke by the European Union. The method was successfully applied for the analysis of commercial samples.

Xylanase production by Aureobasidium pullulans on globe artichoke stem: Bioprocess optimization, enzyme characterization and application in saccharification of lignocellulosic biomass.

Statistical optimization of the factors affecting xylanase production by Aureobasidium pullulans NRRL Y-2311-1 on globe artichoke stem was performed for the first time. The optimization strategies used resulted in almost six-fold enhancement of xylanase production (66.48 U/ml). Biochemical and thermal characterization of the crude xylanase preparation was performed to elucidate its feasibility for different industrial applications. The optimum conditions for xylanase activity were pH 4.0 and 30-50°C. The enzyme was very stable over a wide pH range of 3.0-8.0. The thermal stability studies revealed an inactivation energy of 183 kJ/mol. Thermodynamic parameters (enthalpy, entropy, and Gibbs free energy) for thermal inactivation were also determined. Primary application of the crude xylanase preparation in saccharification of corn cob subjected to different pretreatment techniques has been evaluated. The crude xylanase preparation was very promising for saccharification of corn cob pretreated with aqueous ammonia. The maximum yield of reducing sugar was 357 mg/g dry substrate, which revealed that the crude xylanase from A. pullulans could be a very good alternative in saccharification of lignocellulosic biomass for biological fuel generation. This study also provides a basis for further exploitation of globe artichoke by-products in microbial production of several other industrially significant metabolites.