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  • Nutritional and functional evaluation of inula crithmoides and mesembryanthemum nodiflorum grown in different salinities for human consumption
    Publication . Ribeiro Lima, Alexandre; Gama, Florinda; CastaƱeda-Loaiza, Viana; Costa, Camila; Schueler, Lisa M.; Santos, Tamara; Salazar, Miguel; Nunes, Carla; Cruz, Rui M. S.; Varela, J.; Barreira, Luƭsa
    The nutritional composition and productivity of halophytes is strongly related to the biotic/abiotic stress to which these extremophile salt tolerant plants are subjected during their cultivation cycle. In this study, two commercial halophyte species (Inula crithmoides and Mesembryanthemum nodiflorum) were cultivated at six levels of salinity using a soilless cultivation system. In this way, it was possible to understand the response mechanisms of these halophytes to salt stress. The relative productivity decreased from the salinities of 110 and 200 mmol Lāˆ’1 upwards for I. crithmoides and M. nodiflorum, respectively. Nonetheless, the nutritional profile for human consumption remained balanced. In general, I. crithmoides vitamin (B1 and B6) contents were significantly higher than those of M. nodiflorum. For both species, Ī²-carotene and lutein were induced by salinity, possibly as a response to oxidative stress. Phenolic compounds were more abundant in plants cultivated at lower salinities, while the antioxidant activity increased as a response to salt stress. Sensory characteristics were evaluated by a panel of culinary chefs showing a preference for plants grown at the salt concentration of 350 mmol Lāˆ’1. In summary, salinity stress was effective in boosting important nutritional components in these species, and the soilless system promotes the sustainable and safe production of halophyte plants for human consumption.
  • In vitro enzyme inhibitory and anti-oxidant properties, cytotoxicity and chemical composition of the halophyte Malcolmia littorea (L.) R.Br. (Brassicaceae)
    Publication . CastaƱeda-Loaiza, Viana; Placines, ChloĆ©; Rodrigues, Maria Joao; Pereira, Catarina; Zengin, Gokhan; Neng, Nuno R.; Nogueira, JosĆ© M. F.; CustĆ³dio, LuĆ­sa
    This work reports for the first time the in vitro anti-oxidant (towards DPPH, ABTS, copper and iron), enzymatic inhibitory (on AChE, BuChE, Ī±-glucosidase, Ī±-amylase and tyrosinase), cytotoxicity (towards HepG2 and HEK 293 cells), and metabolomics (by HPLC-MS) of extracts from organs of Malcolmia littorea (L.) R.Br. Extracts were constituted mainly by phenolic acids and flavonoids, and main compounds were salicylic acid and luteolin-7-O-glucoside. Samples showed reduced radical scavenging and metal chelating capacity, and only the methanol extracts reduced iron. The root's ethanol and methanol extracts, and the aerial organ's ethanol extract exhibited the highest AChE inhibition. The root's ethanol extract displayed dual anti-cholinesterase activity. Samples showed a low capacity to inhibit Ī±-amylase, but a high Ī±-glucosidase inhibition was obtained with the root's and flower's ethanol extracts, and flower's methanol extract. Overall, samples displayed a high inhibition against tyrosinase, reduced HepG2 cellular viability and were less toxic towards HEK 293 cells.
  • The irrigation salinity and harvesting affect the growth, chemical profile and biological activities of Polygonum maritimum L.
    Publication . Rodrigues, Maria Joao; Monteiro, Ivo; Placines, ChloĆ©; CastaƱeda-Loaiza, Viana; Slusarczyk, Sylwester; Matkowski, Adam; Pereira, Catarina; Pousao-Ferreira, Pedro; CustĆ³dio, LuĆ­sa
    Previously, our group identified the halophyte Polygonum maritimum L. (sea knotgrass) as a promising source of ingredients for the cosmetic, food, pharmaceutical and veterinarian industries. To further explore this species, and to guarantee the supply of biomass for commercial purposes, it is necessary to ensure its sustainable production while assuring the preservation of its chemical and biological properties. In this context, this work aimed to cultivate this species in greenhouse conditions and to determine the influence of specific agronomic conditions, namely irrigation salinity and harvest, on the plant's growth performance and biological properties of obtained biomass. For that purpose, plants were grown in a greenhouse and irrigated with water with different salinities (0, 100, 200, 300 and 600 mM of sodium chloride - NaCl). After six weeks of irrigation, plants were cut 7 cm above soil level and submitted to an additional two harvests with a six-week interval. Plant growth performance was evaluated in terms of plant height, leaf number and surface area, moisture, and productivity. Acetone extracts were prepared from aboveground organs and evaluated for chemical composition (by spectrophotometric methods, and by ultra-high-resolution mass spectrometry - UHRMS), and for in vitro antioxidant properties [radical-scavenging activity (RSA) on DPPH and ABTS, ferric reducing antioxidant power (FRAP) and metal chelating activity on iron (ICA) and copper (CCA)]. Extracts were also appraised for in vitro anti-inflammatory activity on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Increased salinity and multiple harvests reduced plant growth and yields; the highest productivity was obtained in plants irrigated with freshwater at 2nd harvest (346 g/m(2) of dried biomass). The extracts maintained the in vitro biological properties and interesting chemical profile, however, these depended on the irrigation salinity and harvest regime. The highest antioxidant activities were obtained in extracts from plants irrigated with freshwater at the 3rd harvest (DPPH: 96.2%; ABTS: 89.1%; CCA: 61.6%; FRAP: 136%). The extracts from biomass obtained in 1st the harvest of freshwater-irrigated plants, and from those treated with 100 and 200 mM of NaCl, had significant anti-inflammatory properties. The main compounds detected were mostly flavonols (myricetin and quercetin glycosides), which varied according to both irrigation salinity and harvest. Our results indicate that sea knotgrass can be cultivated in greenhouse conditions aiming industrial commercial applications, irrigated with freshwater or with irrigation solutions with moderate salinity. Moreover, produced biomass maintain the biological and chemical properties previously detected in plants collected from the wild.
  • Dataset on functional and chemical properties of the medicinal halophyte Polygonum maritimum L. under greenhouse cultivation
    Publication . Rodrigues, Maria Joao; Monteiro, Ivo; Placines, ChloĆ©; CastaƱeda-Loaiza, Viana; Ślusarczyk, Sylwester; Matkowski, Adam; Pereira, Catarina; PousĆ£o-Ferreira, Pedro; CustĆ³dio, LuĆ­sa
    This data article includes data and analyses on the effect of different agronomic techniques on the production of Polygonum maritimum L. (sea knotgrass), namely different salinity irrigation treatments (0, 100, 200, 300 and 600 mM of NaCl) and a multi-harvest regime, and their relation with the chemical profile (ultra-high-resolution mass spectrometry - UHRMS), in vitro antioxidant [radical-scavenging activity (RSA) of DPPH and ABTS, copper chelating activity and ferric reducing antioxidant power] and anti-inflammatory (nitric oxide reduction on lipopolysaccharide-stimulated macrophages) activities. For further interpretation of the data presented in this work, please see the related research article "The irrigation salinity and harvesting affect the growth, chemical profile and biological activities of Polygonum maritimum L." (Rodrigues et al., 2019).
  • Wild vs cultivated halophytes: Nutritional and functional differences
    Publication . CastaƱeda-Loaiza, Viana; Oliveira, Marta; Santos, Tamara; SchĆ¼ler, Lisa M.; Ribeiro Lima, Alexandre; Gama, Florinda; Salazar, Miguel; Neng, N.R.; Nogueira, J. M. F.; Varela, JoĆ£o; Barreira, LuĆ­sa
    Some halophyte plants are currently used in gourmet cuisine due to their unique organoleptic properties. Moreover, they exhibit excellent nutritional and functional properties, being rich in polyphenolics and vitamins. These compounds are associated to strong antioxidant activity and enhanced health benefits. This work compared the nutritional properties and antioxidant potential of three species (Mesembryanthemum nodiflorum, Suaeda maritima and Sarcocornia fruticosa) collected in saltmarshes from Portugal and Spain with those of cultivated plants. the latter were generally more succulent and had higher contents of minerals than plants obtained from the wild and contained less fibre. All species assayed are a good source of proteins, fibres and minerals. Additionally, they are good sources of carotenoids and vitamins A, C and B-6 and showed good antioxidant potential particularly S. maritima. Chromatographic analysis of the phenolic profile revealed that ferulic and caffeic acids as the most relevant phenolic compounds detected in the halophytes tested.
  • A review on sarcocornia species: Ethnopharmacology, nutritional properties, phytochemistry, biological activities and propagation
    Publication . CustĆ³dio, LuĆ­sa; Rodrigues, Maria JoĆ£o; Pereira, Catarina Guerreiro; CastaƱeda-Loaiza, Viana; Fernandes, Eliana; Standing, Dominic; Neori, Amir; Shpigel, Muki; Sagi, Moshe
    Sarcocornia A. J. Scott is a halophytic edible succulent plant belonging to the Amaranthaceae family. To date, the genus includes 28 species distributed worldwide in saline environments, usually salt marshes. Sarcocornia (Scott) is similar to Salicornia (L.), which has a recognized commercial value in morphological and taxonomical traits. Species of both genera are commonly named samphire or glassworts in Europe, and their fleshy shoots are commercialized under their traditional names. Due to their nutritional, organoleptic and medicinal properties, Sarcocornia species have a high economic potential in various biotechnology sectors. Being highly tolerant to salt, they can be cultivated in saline conditions, and dissimilar to Salicornia, they are perennial, i.e., they can be harvested year-round. Therefore, Sarcocornia species are considered promising gourmet vegetables to be explored in the context of climate change, soil and water salinization and eco-sustainability. We hereby put together and reviewed the most relevant information on Sarcocornia taxonomy, morphology, nutritional and pharmacological properties, uses in ethnomedicine, potential applications in biotechnology, and propagation strategies.
  • Establishment and elicitation of liquid adventitious root cultures of Inula crithmoides L. for increased caffeoylquinic acids production and hepatoprotective properties
    Publication . CustĆ³dio, LuĆ­sa; CziĆ”ky, ZoltĆ”n; CastaƱeda-Loaiza, Viana; Rodrigues, Maria Joao
    This study established a liquid adventitious root culture for Inula crithmoides L. focusing on producing extracts rich in hepatoprotective caffeoylquinic acids through elicitation. Adventitious roots were induced from in vitro-grown plant leaves cultured on MS medium supplemented with 1 mg/L IBA. Growth and secondary metabolites accumulation (phenolics, flavonoids, hydroxycinnamic acids) were monitored weekly to determine the optimal harvesting time. Then, the adventitious roots were elicited for 4 weeks with varying concentrations of yeast extract-YE (50, 100 and 200 mg/L) and extracted with ethanol. The extracts were evaluated for their hepatoprotective effects against ethanol-induced hepatotoxicity and analyzed through high-performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS/MS). Roots of greenhouse-cultivated plants were used to compare the results obtained from in vitro -grown adventitious roots. IBA supplementation induced 100% adventitious root formation, with maximum biomass and phenolic accumulation after 4 weeks. Hydroxycinnamic acid levels increased significantly with YE treatment at all the concentrations, particularly in case of chlorogenic acid and di-O-caffeoylquinic acid isomers. The root extracts displayed significant in vitro hepatoprotective effects, particularly at 50 mg/L YE elicitation. Overall, the elicitation of I. crithmoides liquid root cultures with yeast extract offers a promising, efficient, and cost-effective approach for optimizing the production of valuable caffeoylquinic acids having potential pharmaceutical use.
  • If you cannot beat them, join them: exploring the fruits of the invasive species Carpobrotus edulis (L.) NE Br as a source of bioactive products
    Publication . CastaƱeda-Loaiza, Viana; Placines, ChloĆ©; Rodrigues, Maria JoĆ£o; Pereira, Catarina; Zengin, Gokhan; Uysal, Ahmet; Jeko, Jozsef; Cziaky, Zoltan; Reis, Catarina Pinto; Gaspar, Maria Manuela; CustĆ³dio, LuĆ­sa
    The halophyte species Carpobrotus edulis (L.) N.E. Br, also known as Hottentot-fig, is one of the 20 most aggressive invasive species of coastal areas worldwide. It is native to South Africa, where it is used in traditional medicine for the treatment of several diseases, including tuberculosis and acquired immunodeficiency syndrome (AIDS) caused by the human immunodeficiency virus (HIV). Aiming at a sustainable use of its biomass as a value-added product, this work reports for the first time the in vitro antioxidant, anti-microbial, enzymatic inhibitory properties and toxicity of peel and flesh extracts of Hottentot-fig mature fruits. The extracts' chemical composition was also determined by spectrophotometric methods (total contents of phenolics: TPC; flavonoids: TFC and tannins: TTC), and by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS/MS). The peels' extracts had generally the highest TPC, TFC and TTC, especially the ethanol ones (TPC: 272.82 mg gallic acid equivalents (GAE)/g dry weight (DW), TFC: 1.58 mg quercetin equivalents (QE)/g DW and TTC: 20.3 mg catechin equivalents (CE)/g DW). The peels' extracts also had the highest diversity of compounds, mostly phenolic acids, flavonoids, and coumarins, as identified by HPLC-ESI-MS/MS. Some molecules were specific to a particular fruit part, for example, coumaric acid and uvaol in the peel, and vanillin and kaempferol-O-(rhamnosyl)hexosylhexoside in the flesh. Some compounds are here described for the first time in Hottentot-fig, such as azelaic acid and emodin. The peel's extracts had the highest anti radical activity, especially the ethanol and acetone towards 2,2-diphenyl-1-picrylhydrazyl (DPPH) (half maximal inhibitory concentration (IC50) values of 0.59 and 0.88 mg/mL, respectively), and the acetone extract against 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) (IC50 = 0.56 mg/mL). Samples had nil capacity to chelate iron, a low copper chelation potential, but a significant capacity to reduce iron, especially the ethanol (IC50 = 0.09 mg/mL) and the acetone extracts of peels (IC50 = 0.10 mg/mL) and flesh (IC50 = 0.11 mg/mL) and also the water peel's extracts (IC50 = 0.18 mg/mL). Samples had nil to low activity towards the enzymes acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), alpha-amylase and alpha-glucosidase, but displayed a strong inhibition of tyrosinase, especially the ethanol peel's extracts (29.55 mg kojic acid equivalents (KAE)/g). Samples had nil to low in vitro toxicity towards human keratinocytes. All together our results suggests possible novel biotechnological applications of Hottentot-fig fruits as sources of innovative bioactive ingredients for the food, cosmetic, agriculture and/or pharmaceutical industries.
  • A first approach for the micropropagation of the medicinal halophyte Polygonum maritimum L. and phenolic profile of acclimatized plants
    Publication . CustĆ³dio, LuĆ­sa; Slusarczyk, Sylwester; Matkowski, Adam; CastaƱeda-Loaiza, Viana; Fernandes, Eliana; Pereira, Catarina; Rodrigues, Maria JoĆ£o
    Polygonum maritimum L. (sea knotgrass) belongs to a genus commonly used in folk medicine to treat inflammation-related disorders. In vitro pharmacological studies have confirmed these properties that were ascribed to bioactive flavonoids, such as myricetin and quercetin glycosides. Therefore, this study aimed at establishing a micropropagation procedure for sea knotgrass for obtaining standardized materials for its potential commercial cultivation. For that, a complete plant regeneration protocol was developed by improving shoot multiplication from nodal explants, rooting and acclimatization procedures, followed by the assessment of the phenolic profile of the in vitro-produced plants. The combination of 3 mg/L 6-benzylaminopurine (BA) + 0.1 mg/L indole-3-acetic acid (IAA) induced the maximum shoot formation (10.3), which was significantly increased from the first to the second cycle (18.3). The best rooting capacity was observed on shoots derived from the control medium (100%), followed by 2 mg/L kinetin (KIN) (97%) and 3 mg/L BA + 0.1 mg/L IAA (90%); however, the shoot number at the end of the rooting phase was higher on shoots derived from 3 mg/L BA + 0.1 mg/L IAA (6.16). The plant growth regulators used in the multiplication phase influenced survival in the acclimatization process, and plants derived from the control medium had the highest survival percentage (63.1%). Acetone extracts made from aerial organs of micropropagated sea knotgrass showed a predominance of the flavonoid myricetin-3-O-rhamnoside (8.135 mg/g). Overall, the halophyte sea knotgrass was successfully micropropagated showing its potential as a medicinal crop for the extraction of bioactive molecules.
  • Growth performance, in vitro antioxidant properties and chemical composition of the halophyte Limonium algarvense Erben are strongly influenced by the irrigation salinity
    Publication . Rodrigues, Maria Joao; Monteiro, Ivo; CastaƱeda-Loaiza, Viana; Placines, ChloĆ©; ConceiĆ§Ć£o Oliveira, M.; Reis, Catarina; Caperta, Ana D.; Soares, Florbela; PousĆ£o-Ferreira, Pedro; Pereira, Catarina; CustĆ³dio, LuĆ­sa
    Limonium algarvense Erben (sea lavender) is a halophyte species with potential to provide natural ingredients with in vitro antioxidant, anti-inflammatory, neuroprotective and antidiabetic properties. This study reports for the first time the 1) cultivation of sea lavender in greenhouse conditions under irrigation with freshwater (approx. 0 mM NaCl) and saline aquaculture wastewater (300 and 600 mM NaCl), and 2) the influence of the irrigation salinity on the plant performance (e.g growth, number of produced leaves and flowers), in vitro antioxidant properties [radical scavenging activity (DPPH and ABTS), ferric reducing antioxidant power (FRAP), metal chelating properties on copper (CCA) and iron (ICA)], toxicity (in vitro on three mammalian cell lines) and chemical composition (determined by LC-ESI-HRMS/MS). The freshwater-irrigated plants had better growth performance than those irrigated with saltwater. Extracts from wild plants, had the highest antioxidant activity, but those from cultivated ones kept high in vitro antioxidant properties and interesting chemical profile. The flowers' extracts of plants irrigated with 300 mM NaCl had the highest antioxidant activities against DPPH, whereas those from freshwater-irrigated plants were more active on ABTS, CCA and FRAP. Most of the extracts showed nil toxicity. The flowers' extracts displayed the highest diversity of compounds, mainly quercetin, apigenin, luteolin, naringenin and their glycoside derivatives. Moreover, their abundance varied with the irrigation salinity. These data indicate that sea lavender plants can be successfully cultivated in greenhouse conditions under fresh- and saltwater irrigation, maintaining interesting biological and chemical properties.