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The xyloglucan endotransglucosylase/hydrolases (XTHs) are enzymes involved in cell wall assembly and growth regulation, cleaving and re-joining hemicellulose chains in the xyloglucan-cellulose network. Here, in a homologous system, we compare the secretion patterns of XTH11, XTH33 and XTH29, three members of the Arabidopsis thaliana XTH family, selected for the presence (XTH11 and XTH33) or absence (XTH29) of a signal peptide, and the presence of a transmembrane domain (XTH33). We show that XTH11 and XTH33 reached, respectively, the cell wall and plasma membrane through a conventional protein secretion (CPS) pathway, whereas XTH29 moves towards the apoplast following an unconventional protein secretion (UPS) mediated by exocyst-positive organelles (EXPOs). All XTHs share a common C-terminal functional domain (XET-C) that, for XTH29 and a restricted number of other XTHs (27, 28 and 30), continues with an extraterminal region (ETR) of 45 amino acids. We suggest that this region is necessary for the correct cell wall targeting of XTH29, as the ETR-truncated protein never reaches its final destination and is not recruited by EXPOs. Furthermore, quantitative real-time polymerase chain reaction analyses performed on 4-week-old Arabidopsis seedlings exposed to drought and heat stress suggest a different involvement of the three XTHs in cell wall remodeling under abiotic stress, evidencing stress-, organ- and time-dependent variations in the expression levels. Significantly, XTH29, codifying the only XTH that follows a UPS pathway, is highly upregulated with respect to XTH11 and XTH33, which code for CPS-secreted proteins.
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Proteínas de Arabidopsis/metabolismo , Parede Celular/metabolismo , Glicosiltransferases/metabolismo , Arabidopsis/metabolismo , Arabidopsis/fisiologia , Proteínas de Arabidopsis/fisiologia , Brefeldina A/farmacologia , Membrana Celular/metabolismo , Desidratação , Glicosiltransferases/fisiologia , Complexo de Golgi/metabolismo , Resposta ao Choque Térmico , Sistemas de Translocação de Proteínas/efeitos dos fármacosRESUMO
The quest to discover the variety of ecological niches inhabited by Saccharomyces cerevisiae has led to research in areas as diverse as wineries, oak trees and insect guts. The discovery of fungal communities in the human gastrointestinal tract suggested the host's gut as a potential reservoir for yeast adaptation. Here, we report the existence of yeast populations associated with the human gut (HG) that differ from those isolated from other human body sites. Phylogenetic analysis on 12 microsatellite loci and 1715 combined CDSs from whole-genome sequencing revealed three subclusters of HG strains with further evidence of clonal colonization within the host's gut. The presence of such subclusters was supported by other genomic features, such as copy number variation, absence/introgressions of CDSs and relative polymorphism frequency. Functional analysis of CDSs specific of the different subclusters suggested possible alterations in cell wall composition and sporulation features. The phenotypic analysis combined with immunological profiling of these strains further showed that sporulation was related with strain-specific genomic characteristics in the immune recognition pattern. We conclude that both genetic and environmental factors involved in cell wall remodelling and sporulation are the main drivers of adaptation in S. cerevisiae populations in the human gut.
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Evolução Molecular , Microbioma Gastrointestinal , Trato Gastrointestinal/microbiologia , Insetos/microbiologia , Saccharomyces cerevisiae/genética , Animais , Variações do Número de Cópias de DNA , Genoma Fúngico , Genômica , Humanos , Microbiota , Repetições de Microssatélites , Filogenia , Saccharomyces cerevisiae/classificação , Saccharomyces cerevisiae/isolamento & purificaçãoRESUMO
BACKGROUND: Table olives have been a component of the Mediterranean diet for centuries, with the trend for their consumption currently increasing worldwide. They are rich in bioactive molecules with nutritional, antioxidant, anti-inflammatory or hormone-like properties. In the present study, the concentrations of phenolics, triterpenic acids, carotenoids and vitamins, as well as fatty acid profiles and antioxidant activity, were analyzed in the edible portion of black table olives (Olea europea L.) from Italian (Cellina di Nardò and Leccino) and Greek (Kalamàta and Conservolea) cultivars fermented with selected autochthonous starters and in the corresponding monovarietal olive oils. RESULTS: On a fresh weight basis, Cellina di Nardò and Leccino table olives showed the highest total phenolic content. No significant differences were found with respect to the levels of total triterpenic (maslinic and oleanolic) acids and vitamin E among cultivars. All table olives were characterized by high amounts of oleic, linoleic and palmitic acids. Oils were richer in lipophilic antioxidants (carotenoids and tocochromanols) than table olives, which, instead, showed a higher content of polyphenols and triterpenic acids than oils. CONCLUSION: The present study demonstrates that fermented table olives are an excellent natural source of unsaturated fatty acids, as well as being nutritionally important health-promoting bioactive compounds. © 2017 Society of Chemical Industry.
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Frutas/química , Lactobacillus plantarum/metabolismo , Olea/microbiologia , Pichia/metabolismo , Saccharomyces cerevisiae/metabolismo , Antioxidantes/análise , Antioxidantes/metabolismo , Ácidos Graxos Insaturados/análise , Ácidos Graxos Insaturados/metabolismo , Fermentação , Frutas/metabolismo , Frutas/microbiologia , Grécia , Itália , Olea/química , Olea/metabolismo , Polifenóis/análise , Polifenóis/metabolismo , Vitamina E/análise , Vitamina E/metabolismoRESUMO
Tomato (Solanum lycopersicon L.) fruit is rich in various nutrients, vitamins and health-promoting molecules. Fresh tomatoes are an important part of the Mediterranean gastronomy, and their consumption is thought to contribute substantially to the reduced incidence of some chronic diseases in the Mediterranean populations in comparison with those of other world areas. Unfortunately, tomato fruit is highly perishable, resulting in important economic losses and posing a challenge to storage, logistic and supply management. This review summarizes the current knowledge on some important health-promoting and eating quality traits of tomato fruits after harvest and highlights the existence of substantial cultivar-to-cultivar variation in the postharvest evolution of the considered traits according to maturity stage at harvest and in response to postharvest manipulations. It also suggests the need for adapting postharvest procedures to the characteristics of each particular genotype to preserve the optimal quality of the fresh product.
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The immune system is essential to maintain the mutualistic homeostatic interaction between the host and its micro- and mycobiota. Living as a commensal,Saccharomyces cerevisiaecould potentially shape the immune response in a significant way. We observed thatS. cerevisiaecells induce trained immunity in monocytes in a strain-dependent manner through enhanced TNFα and IL-6 production upon secondary stimulation with TLR ligands, as well as bacterial and fungal commensals. Differential chitin content accounts for the differences in training properties observed among strains, driving induction of trained immunity by increasing cytokine production and direct antimicrobial activity bothin vitroandin vivo These chitin-induced protective properties are intimately associated with its internalization, identifying a critical role of phagosome acidification to facilitate microbial digestion. This study reveals how commensal and passenger microorganisms could be important in promoting health and preventing mucosal diseases by modulating host defense toward pathogens and thus influencing the host microbiota-immune system interactions.
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Quitina/imunologia , Imunidade Inata , Monócitos/microbiologia , Saccharomyces cerevisiae/imunologia , Animais , Parede Celular/imunologia , Humanos , Interleucina-6/imunologia , Camundongos Endogâmicos C57BL , Monócitos/imunologia , Fagocitose , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Cellulose synthase-like (Csl) genes are believed to encode enzymes for the synthesis of cell wall matrix polysaccharides. The subfamily of CslA is putatively involved in the biosynthesis of ß -mannans. Here we report a study on the cellular localization and the enzyme activity of an Arabidopsis CslA family member, AtCslA2. We show that the fluorescent protein fusion AtCslA2-GFP, transiently expressed in tobacco leaf protoplasts, is synthesized in the ER and it accumulates in the Golgi stacks. The chimera is inserted in the Golgi membrane and is functional since membrane preparations obtained by transformed protoplasts carry out the in vitro synthesis of a 14C-mannan starting from GDP-D-[U-14C]mannose as substrate. The enzyme specific activity is increased by approximately 38% in the transformed protoplasts with respect to wild-type. Preliminary tests with proteinase K, biochemical data, and TM domain predictions suggest that the catalytic site of AtCslA2 faces the Golgi lumen.
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Arabidopsis , Quimera , Corantes Fluorescentes/química , Glucosiltransferases/química , Rede trans-Golgi/química , Arabidopsis/enzimologia , Fenômenos Bioquímicos , Quimera/metabolismo , Corantes Fluorescentes/metabolismo , Glucosiltransferases/metabolismo , Folhas de Planta/enzimologia , Nicotiana/enzimologia , Rede trans-Golgi/enzimologiaRESUMO
Carotenoids are well known for their nutritional properties and health promoting effects representing attractive ingredients to develop innovative functional foods, nutraceutical and pharmaceutical preparations. Pumpkin (Cucurbita spp.) flesh has an intense yellow/orange color owing to the high level of carotenoids, mainly α-carotene, ß-carotene, ß-cryptoxanthin, lutein and zeaxanthin. There is considerable interest in extracting carotenoids and other bioactives from pumpkin flesh. Extraction procedures able to preserve nutritional and pharmacological properties of carotenoids are essential. Conventional extraction methods, such as organic solvent extraction (CSE), have been used to extract carotenoids from plant material for a long time. In recent years, supercritical carbon dioxide (SC-CO2) extraction has received a great deal of attention because it is a green technology suitable for the extraction of lipophylic molecules and is able to give extracts of high quality and totally free from potentially toxic chemical solvents. Here, we review the results obtained so far on SC-CO2 extraction efficiency and quali-quantitative composition of carotenoids from pumpkin flesh. In particular, we consider the effects of (1) dehydration pre-treatments; (2) extraction parameters (temperature and pressure); the use of water, ethanol and olive oil singularly or in combination as entrainers or pumpkin seeds as co-matrix.
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Dióxido de Carbono/química , Carotenoides/isolamento & purificação , Cucurbita/metabolismo , Cromatografia com Fluido Supercrítico , Pressão , Solventes/química , TemperaturaRESUMO
Grape variety, quality, geographic origins and phytopathology can influence the amount of polyphenols that accumulate in grape tissues. Polyphenols in wine not only shape their organoleptic characteristics but also significantly contribute to the positive impact that this beverage has on human health. However, during the winemaking process, the total polyphenol content is substantially reduced due to the adsorption onto yeast wall polymers and subsequent lees separation. Despite this, limited information is available regarding the influence of the yeast starter strain on the polyphenolic profile of wine. To address this issue, a population consisting of 136 Saccharomyces cerevisiae strains was analyzed to identify those with a diminished ability to adsorb polyphenols. Firstly, the reduction in concentration of polyphenolic compounds associated to each strain was studied by assaying Total Phenolic Content (TPC) and Trolox Equivalent Antioxidant Capacity (TEAC) in the wines produced by micro-scale must fermentation. A total of 29 strains exhibiting a TPC and TEAC reduction ≤ 50%, when compared to that detected in the utilized grape must were identified and the nine most-promising strains were further validated by larger-scale vinification. Physico-chemical analyses of the resulting wines led to the identification of four strains, namely ITEM6920, ITEM9500, ITEM9507 and ITEM9508 which showed, compared to the control wine, a TPC and TEAC reduction ≤ 20 in the produced wines. They were denoted by a significant (p < 0.05) increased amount of anthocyanin, quercetin and trans-coutaric acid, minimal volatile acidity (<0.2 g/L), absence of undesirable metabolites and a well-balanced volatile profile. As far as we know, this investigation represents the first clonal selection of yeast strains aimed at the identifying "functional" fermentation starters, thereby enabling the production of regional wines with enriched polyphenolic content.
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BACKGROUND: Many fruits, including watermelon, are proficient in carotenoid accumulation during ripening. While most genes encoding steps in the carotenoid biosynthetic pathway have been cloned, few transcriptional regulators of these genes have been defined to date. Here we describe the identification of a set of putative carotenoid-related transcription factors resulting from fresh watermelon carotenoid and transcriptome analysis during fruit development and ripening. Our goal is to both clarify the expression profiles of carotenoid pathway genes and to identify candidate regulators and molecular targets for crop improvement. RESULTS: Total carotenoids progressively increased during fruit ripening up to ~55 µg g(-1) fw in red-ripe fruits. Trans-lycopene was the carotenoid that contributed most to this increase. Many of the genes related to carotenoid metabolism displayed changing expression levels during fruit ripening generating a metabolic flux toward carotenoid synthesis. Constitutive low expression of lycopene cyclase genes resulted in lycopene accumulation. RNA-seq expression profiling of watermelon fruit development yielded a set of transcription factors whose expression was correlated with ripening and carotenoid accumulation. Nineteen putative transcription factor genes from watermelon and homologous to tomato carotenoid-associated genes were identified. Among these, six were differentially expressed in the flesh of both species during fruit development and ripening. CONCLUSIONS: Taken together the data suggest that, while the regulation of a common set of metabolic genes likely influences carotenoid synthesis and accumulation in watermelon and tomato fruits during development and ripening, specific and limiting regulators may differ between climacteric and non-climacteric fruits, possibly related to their differential susceptibility to and use of ethylene during ripening.
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Carotenoides/genética , Citrullus/crescimento & desenvolvimento , Frutas/crescimento & desenvolvimento , Redes e Vias Metabólicas/genética , Carotenoides/metabolismo , Citrullus/efeitos dos fármacos , Citrullus/metabolismo , Etilenos/farmacologia , Frutas/efeitos dos fármacos , Frutas/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Licopeno , Redes e Vias Metabólicas/efeitos dos fármacos , Proteínas de Plantas/biossíntese , Plantas Geneticamente ModificadasRESUMO
Strain SPC-1(T) was isolated from the phyllosphere of Cynara cardunculus L. var. sylvestris (Lamk) Fiori (wild cardoon), a Mediterranean native plant considered to be the wild ancestor of the globe artichoke and cultivated cardoon. This Gram-stain-negative, catalase-positive, oxidase-negative, non-spore-forming, rod-shaped and non-motile strain secreted copious amounts of an exopolysaccharide, formed slimy, viscous, orange-pigmented colonies and grew optimally at around pH 6.0-6.5 and 26-30 °C in the presence of 0-0.5 % NaCl. Phylogenetic analysis based on comparisons of 16S rRNA gene sequences demonstrated that SPC-1(T) clustered together with species of the genus Sphingomonas sensu stricto. The G+C content of the DNA (66.1 mol%), the presence of Q-10 as the predominant ubiquinone, sym-homospermidine as the predominant polyamine, 2-hydroxymyristic acid (C(14 : 0) 2-OH) as the major hydroxylated fatty acid, the absence of 3-hydroxy fatty acids and the presence of sphingoglycolipid supported this taxonomic position. 16S rRNA gene sequence analysis showed that SPC-1(T) was most closely related to Sphingomonas hankookensis ODN7(T), Sphingomonas insulae DS-28(T) and Sphingomonas panni C52(T) (98.19, 97.91 and 97.11 % sequence similarities, respectively). However, DNA-DNA hybridization analysis did not reveal any relatedness at the species level. Further differences were apparent in biochemical traits, and fatty acid, quinone and polyamine profiles leading us to conclude that strain SPC-1(T) represents a novel species of the genus Sphingomonas, for which the name Sphingomonas cynarae sp. nov. is proposed; the type strain is SPC-1(T) ( = JCM 17498(T) = ITEM 13494(T)). A component analysis of the exopolysaccharide suggested that it represents a novel type of sphingan containing glucose, rhamnose, mannose and galactose, while glucuronic acid, which is commonly found in sphingans, was not detected.
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Cynara/microbiologia , Filogenia , Polissacarídeos Bacterianos/biossíntese , Sphingomonas/classificação , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/análise , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Poliaminas/análise , RNA Ribossômico 16S/genética , Sphingomonas/genética , Sphingomonas/isolamento & purificação , Ubiquinona/análiseRESUMO
Supercritical fluid extraction with CO2 (SFE) is an alternative technology to conventional solvent extraction (CSE), to obtain food-grade bioactives from plants. Here, SFE and CSE extracts from carrot and pumpkin matrices, impregnated with hempseed or flaxseed oil as co-solvents, were characterized by HPLC and GC-MS, and their ability to counteract the inflammatory and oxidative phenomena underlying the onset of several pathologies was assessed in vitro. All extracts showed dose-dependent anti-inflammatory potential and demonstrated an ability to interfere with the pro-inflammatory effects of breast cancer cell-conditioned media, and to inhibit reactive oxygen species (ROS) accumulation and nitrite production (NP) in lipopolysaccharide-stimulated macrophages. Nuclear factor-erythroid-2-related factor 2 (Nrf2) is involved in these response mechanisms, as highlighted by the increased mRNA levels of its target genes revealed by quantitative real-time PCR analyses. NP and ROS concentrations negatively correlated with α-tocopherol and most carotenoids, but positively with the total tocopherol/total carotenoid ratio, suggesting an idiosyncratic effect of these bioactives on cell responses and emphasizing the need to focus on extract constituents' interactions.
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Cucurbita , Daucus carota , Animais , Camundongos , Lipopolissacarídeos/farmacologia , Dióxido de Carbono/farmacologia , Meios de Cultivo Condicionados/farmacologia , Espécies Reativas de Oxigênio , Células MDA-MB-231 , Extratos Vegetais/farmacologia , Macrófagos , Estresse Oxidativo , Inflamação/tratamento farmacológico , Carotenoides/farmacologia , Células RAW 264.7RESUMO
Growing conditions and seasonal fluctuations are critical factors affecting fruit and vegetable nutritional quality. The effects of two partially overlapping cropping seasons, early (ECS; January-May) and full (FCS; March-July), on the main carpometric traits and bioactive components of different watermelon fruits were investigated in the open field. Four watermelon genotypes, comprising of three commercial cultivars 'Crimson Sweet', 'Dumara', 'Giza', and the novel hybrid 'P503 F1', were compared. The carpometric traits varied significantly between genotypes. Soluble solids and yield were higher under FCS than ECS. The variation affecting colour indexes between the two growing seasons exhibited a genotype-dependent trend. The antioxidant components and radical scavenging activity of watermelon fruits were also significantly affected by differences in received solar energy and temperature fluctuations during the trial period. The average citrulline, total phenolics and flavonoid contents were 93%, 71% and 40% higher in FCS than in ECS. A genotype-dependent variation trend was also observed for lycopene and total vitamin C between cropping seasons. The hydrophilic and lipophilic radical scavenging activities of the pulp of ripe watermelon fruits of the different genotypes investigated varied between 243.16 and 425.31 µmol Trolox Equivalent (TE) of 100 g-1 of fresh weight (fw) and from 232.71 to 341.67 µmol TE of 100 g-1 fw in FCS and ECS, respectively. Our results, although preliminary, show that the functional quality of watermelon fruits is drastically altered depending on the environmental conditions that characterize the ECS and LCS.
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The secretory pathway in plants involves sustained traffic to the cell wall, as matrix components, polysaccharides and proteins reach the cell wall through the endomembrane system. We studied the secretion pattern of cell-wall proteins in tobacco protoplasts and leaf epidermal cells using fluorescent forms of a pectin methylesterase inhibitor protein (PMEI1) and a polygalacturonase inhibitor protein (PGIP2). The two most representative protein fusions, secGFP-PMEI1 and PGIP2-GFP, reached the cell wall by passing through ER and Golgi stacks but using distinct mechanisms. secGFP-PMEI1 was linked to a glycosylphosphatidylinositol (GPI) anchor and stably accumulated in the cell wall, regulating the activity of the endogenous pectin methylesterases (PMEs) that are constitutively present in this compartment. A mannosamine-induced non-GPI-anchored form of PMEI1 as well as a form (PMEI1-GFP) that was unable to bind membranes failed to reach the cell wall, and accumulated in the Golgi stacks. In contrast, PGIP2-GFP moved as a soluble cargo protein along the secretory pathway, but was not stably retained in the cell wall, due to internalization to an endosomal compartment and eventually the vacuole. Stable localization of PGIP2 in the wall was observed only in the presence of a specific fungal endopolygalacturonase ligand in the cell wall. Both secGFP-PMEI1 and PGIP2-GFP sorting were distinguishable from that of a secreted GFP, suggesting that rigorous and more complex controls than the simple mechanism of bulk flow are the basis of cell-wall growth and differentiation.
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Parede Celular/metabolismo , Nicotiana/metabolismo , Proteínas de Plantas/metabolismo , Poligalacturonase/metabolismo , Biomarcadores , Membrana Celular/metabolismo , Retículo Endoplasmático/metabolismo , Endossomos/metabolismo , Proteínas Fúngicas/antagonistas & inibidores , Proteínas Fúngicas/metabolismo , Proteínas Ligadas por GPI/metabolismo , Complexo de Golgi/metabolismo , Hexosaminas/efeitos adversos , Hexosaminas/metabolismo , Mutação , Proteínas de Plantas/farmacologia , Plantas Geneticamente Modificadas , Poligalacturonase/antagonistas & inibidores , Ligação Proteica , Estabilidade Proteica , Transporte Proteico , Protoplastos/metabolismo , Proteínas Qa-SNARE/genética , Proteínas Qa-SNARE/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes de Fusão/farmacologia , Nicotiana/citologia , Nicotiana/genéticaRESUMO
This study aims to describe and compare the distribution of bioactive compounds, the fatty acids profiles, and the TEAC hydrophilic and lipophilic antioxidant activities in different fruit fractions (pulp, peel, and kernel) of two mango cultivars (Tommy Atkins and Keitt). All fractions are sources of health-promoting bioactive compounds. Regardless of cultivars, pulp had the highest content of phytosterols (~150 mg/100 g dw), peels ranked first for pentaciclic triterpenes (from 14.2 to 17.7 mg/100 g dw), tocopherols, carotenoids, and chlorophylls, and kernels for phenolic compounds (from 421.6 to 1464.8 mg/100 g dw), flavonoids, condensed tannins, as well as hydrophilic and lipophilic antioxidant activities. Differences between the two cultivars were evidenced for ascorbic acid, which showed the highest levels in the peels and kernels of Keitt and Tommy Atkins fruits, respectively. Similarly, the concentration of dehydroascorbic acid was higher in the pulp of Tommy Atkins than Keitt. The highest percentage of saturated fatty acids was observed in pulp (~42%) and kernels (~50%), monounsaturated fatty acids in kernels (up to 41%), and polyunsaturated fatty acids in peels (up to 52%). Our results add information to the current knowledge on nutraceuticals' distribution in different fractions of mango fruit, supporting its consumption as a healthy fruit and suggesting the great potential value of peels and kernels as sources of novel ingredients. Indeed, mango by-products generated during agronomic-to-industrial processing not only causes a significant environmental impact, but economic losses too. In this scenario, boosting research on conventional recovery methods offers eco-friendly solutions. However, green, novel biorefinery technologies may offer eco-friendly and profitable solutions, allowing the recovery of several more profitable by-products, sustaining their continuous growth since many bioactive compounds can be recovered from mango by-products that are potentially useful in the design of innovative nutraceutical, cosmeceutical, and pharmaceutical formulations.
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Lycopene content in tomato fruit is largely under genetic control and varies greatly among genotypes. Continued improvement of lycopene content in elite varieties with conventional breeding has become challenging, in part because little is known about the underlying molecular mechanisms in high-lycopene tomatoes (HLYs). We collected 42 HLYs with different genetic backgrounds worldwide. High-performance liquid chromatography (HPLC) analysis revealed lycopene contents differed among the positive control wild tomato Solanum pimpinellifolium, HLYs, the normal lycopene cultivar "Moneymaker", and the non-lycopene cultivar NC 1Y at the pink and red ripe stages. Real-time RT-PCR analysis of expression of the 25 carotenoid biosynthesis pathway genes of each genotype showed a significantly higher expression in nine upstream genes (GGPPS1, GGPPS2, GGPPS3, TPT1, SSU II, PSY2, ZDS, CrtISO and CrtISO-L1 but not the well-studied PSY1, PDS and Z-ISO) at the breaker and/or red ripe stages in HLYs compared to Moneymaker, indicating a higher metabolic flux flow into carotenoid biosynthesis pathway in HLYs. Further conversion of lycopene to carotenes may be prevented via the two downstream genes (ß-LCY2 and ε-LCY), which had low-abundance transcripts at either or both stages. Additionally, the significantly higher expression of four downstream genes (BCH1, ZEP, VDE, and CYP97C11) at either or both ripeness stages leads to significantly lower fruit lycopene content in HLYs than in the wild tomato. This is the first systematic investigation of the role of the complete pathway genes in regulating fruit lycopene biosynthesis across many HLYs, and enables tomato breeding and gene editing for increased fruit lycopene content.
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Improved cellulose biosynthesis and plant biomass represent important economic targets for several biotechnological applications including bioenergy and biofuel production. The attempts to increase the biosynthesis of cellulose by overexpressing CesAs proteins, components of the cellulose synthase complex, has not always produced consistent results. Analyses of morphological and molecular data and of the chemical composition of cell walls showed that tobacco plants (F31 line), stably expressing the Arabidopsis CesA6 fused to GFP, exhibits a "giant" phenotype with no apparent other morphological aberrations. In the F31 line, all evaluated growth parameters, such as stem and root length, leaf size, and lignified secondary xylem, were significantly higher than in wt. Furthermore, F31 line exhibited increased flower and seed number, and an advance of about 20 days in the anthesis. In the leaves of F31 seedlings, the expression of primary CesAs (NtCesA1, NtCesA3, and NtCesA6) was enhanced, as well as of proteins involved in the biosynthesis of non-cellulosic polysaccharides (xyloglucans and galacturonans, NtXyl4, NtGal10), cell wall remodeling (NtExp11 and XTHs), and cell expansion (NtPIP1.1 and NtPIP2.7). While in leaves the expression level of all secondary cell wall CesAs (NtCesA4, NtCesA7, and NtCesA8) did not change significantly, both primary and secondary CesAs were differentially expressed in the stem. The amount of cellulose and matrix polysaccharides significantly increased in the F31 seedlings with no differences in pectin and hemicellulose glycosyl composition. Our results highlight the potentiality to overexpress primary CesAs in tobacco plants to enhance cellulose synthesis and biomass production.
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Carotenoids may have different effects on cancer and its progression. The safety of carotenoid supplements was evaluated in vitro on human non-small cell lung cancer (NSCLC) adenocarcinoma A549 cells by the administration of three different oleoresins containing lycopene and other lipophilic phytochemicals, such as tocochromanols. The oleoresins, obtained by the supercritical CO2 green extraction technology from watermelon (Lyc W), gac(Lyc G) and tomato (Lyc T) and chlatrated in α-cyclodextrins, were tested in comparison to synthetic lycopene (Lyc S), by cell cycle, Annexin V-FITC/PI, clonogenic test, Mytosox, intracellular ROS, Western Blot for NF-kB and RT-PCR and ELISA for IL-8. The extracts administered at the same lycopene concentration (10 µM) showed conflicting behaviors: Lyc W, with the highest lycopene/tocochromanols ratio, significantly increased cell apoptosis, mitochondrial stress, intracellular ROS, NF-kB and IL-8 expression and significantly decreased cell proliferation, whereas Lyc G and Lyc T significantly increased only cell proliferation. Lyc S treatment was ineffective. The highest amount of lycopene in Lyc W was able to counteract and revert the cell survival effect of tocochromanols supporting the importance of evaluating the lycopene bio-availability and the real effect of antioxidant tocochromanols' supplementation which may not only have no anticancer benefits but may even increase cancer aggressivity.
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The results showed that soil electrical conductivity, (EC2: 7 dS/m) increased soluble solids, lycopene content, total phenolic content, hydrophilic and lipophilic radical scavenging activities (HRSA and LRSA) by 14.2, 149, 20, 46.4, and 19.0%, respectively, compared with control. Under 0.5% spent engine oil (SEO), flavonoid content decreased by 21.7% compared with the control. HRSA and LRSA of fruits subjected to EC2/SEO1 treatment were, respectively, 45.9 and 35.5% lower than control. The a*/b* ratio was positively and significantly (P < 0.01) correlated with ß-carotene (R = 0.78), lycopene (R = 0.68), total vitamin C (R = 0.71), α-tocopherol (R = 0.83), γ-tocopherol (R = 0.66), HRSA (R = 0.93), LRSA (R = 0.80), and soluble solids (R = 0.84) suggesting that it may be a promising indicator of fruit quality in areas affected by such constraints. The research revealed that combined stresses induce responses markedly different from those of individual treatments, which strain the need to focus on how the interaction between stresses may affect the functional quality of tomato fruits.
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Oleosin, caleosin and steroleosin are normally expressed in developing seed cells and are targeted to oil bodies. In the present work, the cDNA of each gene tagged with fluorescent proteins was transiently expressed into tobacco protoplasts and the fluorescent patterns observed by confocal laser scanning microscopy. Our results indicated clear differences in the endocellular localization of the three proteins. Oleosin and caleosin both share a common structure consisting of a central hydrophobic domain flanked by two hydrophilic domains and were correctly targeted to lipid droplets (LD), whereas steroleosin, characterized by an N-terminal oil body anchoring domain, was mainly retained in the endoplasmic reticulum (ER). Protoplast fractionation on sucrose gradients indicated that both oleosin and caleosin-green fluorescent protein (GFP) peaked at different fractions than where steroleosin-GFP or the ER marker binding immunoglobulin protein (BiP), were recovered. Chemical analysis confirmed the presence of triacylglycerols in one of the fractions where oleosin-GFP was recovered. Finally, only oleosin- and caleosin-GFP were able to reconstitute artificial oil bodies in the presence of triacylglycerols and phospholipids. Taken together, our results pointed out for the first time that leaf LDs can be separated by the ER and both oleosin or caleosin are selectively targeted due to the existence of selective mechanisms controlling protein association with these organelles.
Assuntos
Corpos de Inclusão/metabolismo , Nicotiana/metabolismo , Folhas de Planta/metabolismo , Óleos de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Protoplastos/metabolismo , Sementes/metabolismo , Transporte Proteico/fisiologiaRESUMO
The increasing popularity of pomegranate (Punica granatum L.), driven by the awareness of its nutraceutical properties and excellent environmental adaptability, is promoting a global expansion of its production area. This investigation reports the variability in the weight, moisture, pH, total soluble solids, carbohydrates, organic acids, phenolic compounds, fatty acids, antioxidant activities, and element composition of different fruit parts (juices, peels, and kernels) from four (Ako, Emek, Kamel, and Wonderful One) of the most widely cultivated Israeli pomegranate varieties in Salento (South Italy). To the best of our knowledge, this is the first systematic characterization of different fruit parts from pomegranate cultivars grown simultaneously in the same orchard and subjected to identical agronomic and environmental conditions. Significant genotype-dependent variability was observed for many of the investigated parameters, though without any correlation among fruit parts. The levels of phenols, flavonoids, anthocyanins, and ascorbic and dehydroascorbic acids of all samples were higher than the literature-reported data, as was the antioxidant activity. This is likely due to positive interactions among genotypes, the environment, and good agricultural practices. This study also confirms that pomegranate kernels and peels are, respectively, rich sources of punicic acid and phenols together, with several other bioactive molecules. However, the variability in their levels emphasizes the need for further research to better exploit their agro-industrial potential and thereby increase juice-production chain sustainability. This study will help to assist breeders and growers to respond to consumer and industrial preferences and encourage the development of biorefinery strategies for the utilization of pomegranate by-products as nutraceuticals or value-added ingredients for custom-tailored supplemented foods.