The Chloroplast Genome of Endive (Cichorium endivia L.): Cultivar Structural Variants and Transcriptome Responses to Stress Due to Rain Extreme Events.

The chloroplast (cp) genome diversity has been used in phylogeny studies, breeding, and variety protection, and its expression has been shown to play a role in stress response. Smooth- and curly-leafed endives (Cichorium endivia var. latifolium and var. crispum) are of nutritional and economic importance and are the target of ever-changing breeding programmes. A reference cp genome sequence was assembled and annotated (cultivar 'Confiance'), which was 152,809 base pairs long, organized into the angiosperm-typical quadripartite structure, harboring two inverted repeats separated by the large- and short- single copy regions. The annotation included 136 genes, 90 protein-coding genes, 38 transfer, and 8 ribosomal RNAs and the sequence generated a distinct phyletic group within Asteraceae with the well-separated C. endivia and intybus species. SSR variants within the reference genome were mostly of tri-nucleotide type, and the cytosine to uracil (C/U) RNA editing recurred. The cp genome was nearly fully transcribed, hence sequence polymorphism was investigated by RNA-Seq of seven cultivars, and the SNP number was higher in smooth- than curly-leafed ones. All cultivars maintained C/U changes in identical positions, suggesting that RNA editing patterns were conserved; most cultivars shared SNPs of moderate impact on protein changes in the ndhD, ndhA, and psbF genes, suggesting that their variability may have a potential role in adaptive response. The cp transcriptome expression was investigated in leaves of plants affected by pre-harvest rainfall and rainfall excess plus waterlogging events characterized by production loss, compared to those of a cycle not affected by extreme rainfall. Overall, the analyses evidenced stress- and cultivar-specific responses, and further revealed that genes of the Cytochrome b6/f, and PSI-PSII systems were commonly affected and likely to be among major targets of extreme rain-related stress.

Quality and Safety of Ready-to-Eat Golden Thistle (Scolymus hispanicus L.): A New Product for Traditional Italian Dishes.

Golden thistle (Scolymus hispanicus L.) is a wild edible plant belonging to Asteraceae family, with a great potential for food applications. The aim of this study was to identify the best cooking procedure able to provide a high-quality, ready-to-use product. For this purpose, leaf midribs (the most used edible part of the plant) were cooked by boiling, steaming, and 'sous vide', and the cooked products were compared for their phenolic content and composition, antioxidant activity, sugar and inorganic ion content, organoleptic characteristics, and microbial safety, this latter also during storage. In general, boiling caused a decrease in the value of these parameters, despite being the best product for taste and overall acceptability. On the contrary, steaming and 'sous vide' resulted in the best treatments to preserve antioxidant activity, total phenols, and chlorogenic acid. In particular, in 'sous vide' cooked samples, a significant increase in the value of these parameters and a remarkable decrease in nitrate content were found. Moreover, 'sous vide' resulted in the best treatment also regarding microbial safety during shelf life; actually, after 15 days of storage at 8 °C, Enterobacteriaceae and mesophilic aerobic bacteria were not detectable in 'sous vide' samples. These results contributed to increase the knowledge of a wild edible plant with high nutritional properties and promoting its consumption by obtaining a ready-to-use product with good organoleptic characteristics and endowed with a long period of shelf life.

Enhancing the nutritional value of Portulaca oleracea L. by using soilless agronomic biofortification with zinc.

Zinc (Zn) plays a crucial role for human health. Zn deficiency is a common problem worldwide, causing health problems specifically related with detrimental effects on immune system. In this study we used hydroponic floating system and nutrient solutions (NS) with different levels of Zn (0.13 - control, 1.3, 2.6 and 5.2 mg/L Zn) in order to test a biofortification process aimed to increase the Zn tissue content of two different varieties (commercial and wild) of purslane. We evaluated the effects of the treatments on yield, visual and overall nutritional quality of the edible part of plants. Biofortification treatments did not affect plant yield, but increased the Zn content in the edible part of purslane by 1.8, 2.3 and 2.7-fold, respectively with 1.3, 2.6 and 5.2 mg/L Zn in the NS, so that the consumption of a serving portion of 150 g of baby leaf purslane biofortified with the highest Zn concentration would account for 21% of RDA (recommended daily allowance) for this nutrient. The Zn biofortification process did not affect colour parameters, therefore no differences in the product visual quality were observed. Zn treatment with 5.2 mg/L allowed to obtain an increase in neoxanthin, lutein and ß-carotene, while it was not possible to outline a common trend for the fatty acids profile in relation to the Zn treatments. The consumption of hydroponic purslane biofortified with Zn may allow to improve the Zn nutritional status of consumers, and provides different important phytochemicals, such as carotenoids and unsaturated fatty acids.

Leaf nutrient content and transcriptomic analyses of endive (Cichorium endivia) stressed by downpour-induced waterlog reveal a gene network regulating kestose and inulin contents.

Endive (Cichorium endivia L.), a vegetable consumed as fresh or packaged salads, is mostly cultivated outdoors and known to be sensitive to waterlogging in terms of yield and quality. Phenotypic, metabolic and transcriptomic analyses were used to study variations in curly- ('Domari', 'Myrna') and smooth-leafed ('Flester', 'Confiance') cultivars grown in short-term waterlog due to rainfall excess before harvest. After recording loss of head weights in all cultivars (6-35%), which was minimal in 'Flester', NMR untargeted profiling revealed variations as influenced by genotype, environment and interactions, and included drop of total carbohydrates (6-50%) and polyols (3-37%), gain of organic acids (2-30%) and phenylpropanoids (98-560%), and cultivar-specific fluctuations of amino acids (-37 to +15%). The analysis of differentially expressed genes showed GO term enrichment consistent with waterlog stress and included the carbohydrate metabolic process. The loss of sucrose, kestose and inulin recurred in all cultivars and the sucrose-inulin route was investigated by covering over 50 genes of sucrose branch and key inulin synthesis (fructosyltransferases) and catabolism (fructan exohydrolases) genes. The lowered expression of a sucrose gene subset together with that of SUCROSE:SUCROSE-1-FRUCTOSYLTRANSFERASE (1-SST) may have accounted for sucrose and kestose contents drop in the leaves of waterlogged plants. Two anti-correlated modules harbouring candidate hub-genes, including 1-SST, were identified by weighted gene correlation network analysis, and proposed to control positively and negatively kestose levels. In silico analysis further pointed at transcription factors of GATA, DOF, WRKY types as putative regulators of 1-SST.

Barattiere: An Italian Local Variety of Cucumis melo L. with Quality Traits between Melon and Cucumber.

Barattiere, belonging to the Cucumis melo L. species, is a local variety of Puglia (Southern Italy), which is consumed as a vegetable at the immature stage, like cucumber. In this study, three Barattiere populations ('Monopoli', 'Carovigno' and 'Fasano') were evaluated for the main quality traits. All genotypes showed a very light green-yellow colour of flesh, without any difference regarding chlorophyll and carotenoid contents. Carovigno's Barattiere showed the highest values of dry weight (6.8 g 100 g-1 fresh weight - FW), sugars (45 g kg-1 FW), and sweetness index (7.3), while Monopoli's Barattiere showed the lowest total phenols content (21 mg kg-1 FW). Fasano's Barattiere showed the highest content of Zn and Cu (2.3 and 0.3 mg kg-1 FW, respectively), while 'Monopoli' showed the highest Ba content (0.3 mg kg-1 FW) and the lowest Mg content (94 mg kg-1 FW). No differences between populations were found concerning the content of Ca, K, Na, B, Mn, and Fe. In conclusion, the quality profile of Barattiere makes this local genotype interesting for its traits, and also suggests its consumption by people with specific dietary requirements.

Morphological and Chemical Profile of Three Tomato (Solanum lycopersicum L.) Landraces of A Semi-Arid Mediterranean Environment.

Puglia (Southern Italy), particularly rich in tomato agro-biodiversity, can be considered a typical region of the semi-arid Mediterranean environments. In this study, three local varieties of tomato (Manduria, Giallo di Crispiano and Regina) were characterized by using morphological descriptors according to international standards. Chemical (isoprenoids, ascorbic acid, total phenols, sugars and mineral content) and agronomic assessment were carried out to highlight the specific traits of these local varieties well adapted to a semi-arid environment. Data of morphological traits according to the "International Union for the Protection of New Varieties of Plants" (UPOV) guideline evidenced a clear distinctness among all three landraces, especially as regards fruits. Results also highlighted that a great part of variation in chemical traits was almost exclusively due to genotypes, while in a few cases observed differences resulted from the interaction between genotype and harvest time. The results of the present study may represent the first step toward the recognition of "conservation variety" status for Regina, Giallo di Crispiano and Manduria tomato landraces. At the same time, both quality traits and agronomic performance of these tomato genotypes suggest the possibility of their cultivation in other semi-arid environments also considering their quality traits, in view of a sustainable production.

Insights into sucrose pathway of chicory stems by integrative transcriptomic and metabolic analyses.

The worldwide-cultivated chicory (Cichorium intybus L.) produces food and beneficial compounds, and young pre-flowering inflorescence stems are newly marketed vegetables. These sink-organs undergo growth by metabolizing sugars of leaf origin; the carbohydrate content and sweetness are crucial aspects for consumers' nutrition and acceptance. NMR profiling of 31 hydrosoluble phytochemicals showed that stem contents varied as influenced by genotype, environment and interaction, and that higher sucrose levels were associated with the sweeter of two landraces. Integrative analyses of metabolic and transcriptomic profile variations allowed the dissection of sucrose pathway. Overall, 427 and 23 unigenes respectively fell into the categories of sucrose metabolism and sugar carriers. Among 10 differentially expressed genes, the 11474/sucrose synthase, 53458/fructokinase, 9306 and 17035/hexokinases, and 20171/SWEET-type genes significantly associated to sugar content variation, and deduced proteins were characterised in silico. Correlation analyses encompassing sugar level variation, expressions of the former genes and of computationally assigned transcription factors (10938/NAC, 14712/bHLH, 40133/TALE and 17846/MIKC) revealed a gene network. The latter was minimally affected by the environment and accomplished with markers, representing a resource for biological studies and breeding.

Iodine Biofortification of Four Brassica Genotypes is Effective Already at Low Rates of Potassium Iodate.

The use of iodine-biofortified vegetables may be a health alternative instead of iodine-biofortified salt for preventing iodine (I) deficiency and related human disorders. In this study, four Brassica genotypes (broccoli raab, curly kale, mizuna, red mustard) were hydroponically grown with three I-IO3- rates (0, 0.75 and 1.5 mg/L) to produce iodine-biofortified vegetables. Crop performances and quality traits were analyzed; iodine content was measured on raw, boiled, and steamed vegetables. The highest I rate generally increased I content in all Brassica genotypes, without plants toxicity effects in terms of reduced growth or morphological symptoms. After 21 day-iodine biofortification, the highest I content (49.5 µg/100 g Fresh Weight (FW)) was reached in broccoli raab shoots, while after 43 day-iodine biofortification, genotype differences were flattened and the highest I content (66 µg/100 g FW, on average) was obtained using 1.5 mg I-IO3/L. Nitrate content (ranging from 1800 to 4575 mg/kg FW) was generally higher with 0.75 mg I-IO3/L, although it depended on genotypes. Generally, boiling reduced iodine content, while steaming increased or left it unchanged, depending on genotypes. Applying low levels of I proved to be suitable, since it could contribute to the partial intake of the recommended dose of 150 µg/day: A serving size of 100 g may supply on average 24% of the recommended dose. Cooking method should be chosen in order to preserve and/or enhance the final I amount.

Transcriptome driven characterization of curly- and smooth-leafed endives reveals molecular differences in the sesquiterpenoid pathway.

Endives (Cichorium endivia L.) are popular vegetables, diversified into curly/frisée- and smooth/broad-leafed (escaroles) cultivar types (cultigroups), and consumed as fresh and bagged salads. They are rich in sesquiterpene lactones (STL) that exert proven function on bitter taste and human health. The assembly of a reference transcriptome of 77,022 unigenes and RNA-sequencing experiments were carried out to characterize the differences between endives and escaroles at the gene structural and expression levels. A set of 3177 SNPs distinguished smooth from curly cultivars, and an SNP-supported phylogenetic tree separated the cultigroups into two distinct clades, consistently with the botanical varieties of origin (crispum and latifolium, respectively). A pool of 699 genes maintained differential expression pattern (core-DEGs) in pairwise comparisons between curly vs smooth cultivars grown in the same environment. Accurate annotation allowed the identification of 26 genes in the sesquiterpenoid biosynthesis pathway, which included several g ermacrene A s ynthase, g ermacrene A o xidase and co stunolide s ynthase members (GAS/GAO/COS module), required for the synthesis of costunolide, a key precursor of lactucopicrin- and lactucin-like sesquiterpene lactones. The core-DEGs contained a GAS gene (contig83192) that was positively correlated with STL levels and recurrently more expressed in curly than smooth endives, suggesting a cultigroup-specific behavior. The significant positive correlation of GAS/GAO/COS transcription and STL abundance (2.4-fold higher in frisée endives) suggested that sesquiterpenoid pathway control occurs at the transcriptional level. Based on correlation analyses, five transcription factors (MYB, MYB-related and WRKY) were inferred to act on contig83192/GAS and specific STL, suggesting the occurrence of two distinct routes in STL biosynthesis.

Efficacy of Combined Sous Vide-Microwave Cooking for Foodborne Pathogen Inactivation in Ready-to-Eat Chicory Stems.

There is a variety of different food processing methods, which can be used to prepare ready-to-eat foods. However, the need to preserve the freshness and nutritional qualities leads to the application of mild technologies which may be insufficient to inactivate microbial pathogens. In this work, fresh chicory stems were packed under a vacuum in films, which were transparent to microwaves. These were then exposed to microwaves for different periods of time. The application of sous vide microwave cooking (SV-MW, 900 W, 2450 MHz), controlled naturally occurring mesophilic aerobic bacteria, yeasts and molds for up to 30 d when vacuum-packed vegetables were stored at 4 °C. In addition, the process lethality of the SV-MW 90 s cooking was experimentally validated. This treatment led to 6.07 ± 0.7 and 4.92 ± 0.65 log cfu/g reduction of Escherichia coli and Listeria monocytogenes inoculated over the chicory stems (100 g), respectively. With an initial load of 9 log cfu/g for both pathogens, less than 10 cfu/g of surviving cells were found after 90 s cooking. This shows that short-time microwave cooking can be used to effectively pasteurize vacuum-packed chicory stems, achieving >5 log cfu/g reduction of E. coli and L. monocytogenes.

Corrigendum: Insights into the Sesquiterpenoid Pathway by Metabolic Profiling and De novo Transcriptome Assembly of Stem-Chicory (Cichorium intybus Cultigroup "Catalogna").

[This corrects the article on p. 1676 in vol. 7, PMID: 27877190.].

Influence of cultivation sites on sterol, nitrate, total phenolic contents and antioxidant activity in endive and stem chicory edible products.

Chicories produce a wide range of vegetables with important nutritional value. We determined the variation of sterol, total polyphenol, nitrate contents and antioxidant capacity (SC, TPC, NC, AC) in endive leaves and stem-chicory novel vegetables, cultivated in two Italian regions. Within a given area, the SC was similar in smooth- and curly leafed endives (106.3-176.0 mg/kg FW); sitosterol and stigmasterol were major fractions (45-56 versus 38-43%). The stem SC was independent of landrace (101.5-118.6 mg/kg FW); sitosterol prevailed on stigmasterol and fucosterol (73-76 versus 12-14% versus 8-9%); the latter reached 15.7 mg/kg FW, conferring value as potential antidiabetes food. The planting site affected the AC and TPC of endives (893.1-1571.4 µmTE/100 g FW, 30.8-76.1 GAE100/g FW) and chicory stems (729.8-1152.5 µmTE/100 g FW; 56.2-124.4 GAE100/g FW), while the NC was recurrently below dangerous thresholds. PCA showed that environment was the major cause of variation, though it modestly affected these parameters.

Insights into the Sesquiterpenoid Pathway by Metabolic Profiling and De novo Transcriptome Assembly of Stem-Chicory (Cichorium intybus Cultigroup "Catalogna").

Stem-chicory of the "Catalogna" group is a vegetable consumed for bitter-flavored stems. Type and levels of bitter sesquiterpene lactones (STLs) participate in conferring bitterness in vegetables. The content of lactucin-and lactucopocrin-like STLs was higher in "Molfettese" than "Galatina" landrace stalks, regardless of the cultivation sites, consistently with bitterness scores and gustative differences. The "Galatina" transcriptome assembly resulted in 58,872 unigenes, 77% of which were annotated, paving the way to molecular investigation of the STL pathway. Comparative transcriptome analysis allowed the identification of 69,352 SNPs and of 1640 differentially expressed genes that maintained the pattern independently of the site. Enrichment analyses revealed that 4 out of 29 unigenes were up-regulated in "Molfettese" vs "Galatina" within the sesquiterpenoid pathway. The expression of two germacrene A -synthase (GAS) and one -oxidase (GAO) genes of the costunolide branch correlated positively with the contents of lactucin-like molecules, supporting that STL biosynthesis regulation occurs at the transcriptional level. Finally, 46 genes encoding transcription factors (TFs) maintained a differential expression pattern between the two varieties regardless of the growth site; correlation analyses among TFs, GAS, GAO gene expressions and STLs contents suggest that one MYB and one bHLH may act in the pathway.

Elemental characterization of wild edible plants from countryside and urban areas.

Thirteen elements (Na, K, Ca, Mg, Fe, Mn, Cu, Zn, Cr, Co, Cd, Ni and Pb) in 11 different wild edible plants (WEP) (Amaranthus retroflexus, Foeniculum vulgare, Cichorium intybus, Glebionis coronaria, Sonchus spp., Borago officinalis, Diplotaxis tenuifolia, Sinapis arvensis, Papaver rhoeas, Plantago lagopus and Portulaca oleracea) collected from countryside and urban areas of Bari (Italy) were determined. B.officinalis and P.rhoeas could represent good nutritional sources of Mn and Fe, respectively, as well as A.retroflexus and S.arvensis for Ca. High intake of Pb and Cd could come from P.lagopus and A.retroflexus (1.40 and 0.13 mg kg(-1) FW, respectively). WEP may give a substantial contribution to the elements intake for consumers, but in some cases they may supply high level of elements potentially toxic for human health. Anyway, both ANOVA and PCA analyses have highlighted the low influence of the harvesting site on the elements content.

Quality evaluation of cook-chilled chicory stems (Cichorium intybus L., Catalogna group) by conventional and sous vide cooking methods.

BACKGROUND: Chicory stems, appreciated both raw and cooked, represent a nutritious and refined food. In this study the effects on the quality of stems cooked by conventional (boiling, steaming and microwaving) and innovative (sous vide) methods were analysed. Several physical, chemical and sensory traits were compared using two local varieties (Galatina and Molfettese) of southern Italy (Puglia region). RESULTS: Independently of the variety, the sous vide method did not significantly affect (redness, yellowness and hue angle) or had the least impact on (lightness and total colour difference) quality parameters among the four methods as compared with the raw product. Following sensory analysis, the sous vide product always showed the highest score among the cooking methods. Moreover, this innovative method did not affect total phenol (TP) content and antioxidant activity (AA) compared with uncooked stems of both varieties. Microwaving increased TP content and AA (though associated with higher weight loss), while different responses depending on the chicory variety were observed after boiling and steaming. CONCLUSION: The results indicate the sous vide technique as optimal to preserve several traits, including organoleptic ones, for the quality of cook-chilled chicory stems. They also provide product-specific information usually required for cooking process strategies in the industrial sector of ready-to-eat vegetables.

[Drop-out from a mental health centre after the first contact]. / Il fenomeno del drop-out precoce dopo il primo contatto in un Centro di Salute Mentale.

AIM: Many patients drop-out from treatment after a single contact; however, characteristics have only rarely been investigated. We aimed at estimating the frequency of early drop-out and identifying possible risk factors. METHODS: The study was performed at the Palestrina Mental Health Centre, located near Rome, Italy. The clinical charts of all first-time attenders who did not come back during a 7-year index period (2000-2006) were reviewed and information was collected on sociodemographic factors, psychiatric history, clinical presentation, and process of care. RESULTS: During the index period, 97 of 1.001 first-time attenders dropped out after the first contact. We excluded patients with symptoms of organic (N = 9) or severe mental disorders (N = 3). Of the remaining 85 patients, 49 presented with symptoms related to life events or situations ("reactive symptoms", RS), while 36 had "non-reactive symptoms" (NRS). Eighty-five randomly selected patients with comparable conditions (psychogenic reaction; anxiety, dysthymic or somatoform disorder; ICD-9 code 300.x, 308.x, 309.x) who came back to visit were included as controls. First-only contact patients tended to present with RS more often (58% vs. 43%) than controls. Therefore, the analyses were stratified for reactivity of symptoms. Among patients with NRS, first-only contact patients tended to be younger than controls. Among patients with RS, time until the next appointment tended to be longer in first-only contact patients than in controls. No other differences were observed between first-only contact patients and controls on sociodemographic variables, source of referral, and previous treatment history. DISCUSSION: The occurrence of early drop-out was relatively low. The higher prevalence of RS among first-only contact patients may suggest a greater disposition towards conflict or ambivalence in interpersonal situations. Patient-perceived service availability, in the form of shorter appointment delay, seems to promote maintenance of contact. Younger patients who tend to be at higher risk of early drop-out deserve increased clinical attention.