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  • A review on sarcocornia species: Ethnopharmacology, nutritional properties, phytochemistry, biological activities and propagation
    Publication . Custódio, Luísa; Rodrigues, Maria João; Pereira, Catarina Guerreiro; Castañeda-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.
  • 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 . Castañeda-Loaiza, Viana; Placines, Chloé; Rodrigues, Maria João; Pereira, Catarina; Zengin, Gokhan; Uysal, Ahmet; Jeko, Jozsef; Cziaky, Zoltan; Reis, Catarina Pinto; Gaspar, Maria Manuela; Custódio, Luísa
    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 . Custódio, Luísa; Slusarczyk, Sylwester; Matkowski, Adam; Castañeda-Loaiza, Viana; Fernandes, Eliana; Pereira, Catarina; Rodrigues, Maria João
    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.
  • Metabolomic profile and biological properties of sea lavender (Limonium algarvense Erben) plants cultivated with aquaculture wastewaters: implications for its use in herbal formulations and food additives
    Publication . Rodrigues, Maria João; Castañeda-Loaiza, Viana; Monteiro, Ivo; Pinela, José; Barros, Lillian; Abreu, Rui M. V.; Oliveira, Maria Conceição; Reis, Catarina; Soares, Florbela; Pousão-Ferreira, Pedro; Pereira, Catarina; Custódio, Luísa
    Water extracts from sea lavender (Limonium algarvense Erben) plants cultivated in greenhouse conditions and irrigated with freshwater and saline aquaculture effluents were evaluated for metabolomics by liquid chromatography-tandem high-resolution mass spectrometry (LC-HRMS/MS), and functional properties by in vitro and ex vivo methods. In vitro antioxidant methods included radical scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), ferric-reducing antioxidant power (FRAP), and copper and iron chelating assets. Flowers’ extracts had the highest compounds’ diversity (flavonoids and its derivatives) and strongest in vitro antioxidant activity. These extracts were further tested for ex vivo antioxidant properties by oxidative haemolysis inhibition (OxHLIA), lipid peroxidation inhibition by thiobarbituric acid reactive substances (TBARS) formation, and anti-melanogenic, anti-tyrosinase, anti-inflammation, and cytotoxicity. Extract from plants irrigated with 300 mM NaCl was the most active towards TBARS (IC50 = 81 µg/mL) and tyrosinase (IC50 = 873 µg/mL). In OxHLIA, the activity was similar for fresh- and saltwater-irrigated plants (300 mM NaCl; IC50 = 136 and 140 µg/mL, respectively). Samples had no anti-inflammatory and anti-melanogenic abilities and were not toxic. Our results suggest that sea lavender cultivated under saline conditions could provide a flavonoid-rich water extract with antioxidant and anti-tyrosinase properties with potential use as a food preservative or as a functional ingredient in herbal supplements.
  • A first approach for the micropropagation of the edible and medicinal Halophyte Inula crithmoides L.
    Publication . Rodrigues, Maria João; Castañeda-Loaiza, Viana; Fernandes, Eliana; Custódio, Luísa
    Inula crithmoides L. (golden samphire) is an edible aromatic halophyte species with confirmed nutritional and medicinal properties attributed to the presence of important metabolites, including proteins, carotenoids, vitamins, and minerals. Therefore, this study aimed at establishing a micropropagation protocol for golden samphire that can serve as a nursery approach to its standardized commercial cultivation. For that purpose, a complete regeneration protocol was developed by improving shoot multiplication from nodal explants, rooting, and acclimatization methodologies. The treatment with BAP alone induced the maximum shoot formation (7–7.8 shoots/explant), while IAA treatment increased the shoot height (9.26–9.5 cm). Furthermore, the treatment that coupled best shoot multiplication (7.8 shoots/explant) and highest shoot height (7.58 cm) was MS medium supplemented with 0.25 mg/L BAP. Moreover, all shoots produced roots (100% rooting), and multiplication treatments did not exert significant effect on root length (7.8–9.7 cm/plantlet). Moreover, by the end of the rooting phase, plantlets cultivated with 0.25 mg/L BAP had the highest shoot number (4.2 shoots/plantlet), and plantlets from 0.6 mg/L IAA + 1 mg/L BAP presented the highest shoot height (14.2 cm) similar to control plantlets (14.0 cm). The survival up to the ex-vitro acclimatization stage was increased from 9.8% (control) to 83.3%, when plants were treated with a paraffin solution. Nevertheless, the in vitro multiplication of golden samphire is a promising way for its rapid propagation and can be used as a nursery method, contributing to the development of this species as an alternative food and medicinal crop.
  • Application of In Vitro plant tissue culture techniques to halophyte species: A review
    Publication . L, Custódio; Charles, Gilbert; Magné, Christian; Barba-Espín, Gregorio; Piqueras, Abel; Hernández, José A.; Ben Hamed, Karim; Castañeda-Loaiza, Viana; Fernandes, Eliana; Rodrigues, Maria João
    Halophytes are plants able to thrive in environments characterized by severe abiotic conditions, including high salinity and high light intensity, drought/flooding, and temperature fluctuations. Several species have ethnomedicinal uses, and some are currently explored as sources of food and cosmetic ingredients. Halophytes are considered important alternative cash crops to be used in sustainable saline production systems, due to their ability to grow in saline conditions where conventional glycophyte crops cannot, such as salt-affected soils and saline irrigation water. In vitro plant tissue culture (PTC) techniques have greatly contributed to industry and agriculture in the last century by exploiting the economic potential of several commercial crop plants. The application of PTC to selected halophyte species can thus contribute for developing innovative production systems and obtaining halophyte-based bioactive products. This work aimed to put together and review for the first time the most relevant information on the application of PTC to halophytes. Several protocols were established for the micropropagation of different species. Various explant types have been used as starting materials (e.g., basal shoots and nodes, cotyledons, epicotyls, inflorescence, internodal segments, leaves, roots, rhizomes, stems, shoot tips, or zygotic embryos), involving different micropropagation techniques (e.g., node culture, direct or indirect shoot neoformation, caulogenesis, somatic embryogenesis, rooting, acclimatization, germplasm conservation and cryopreservation, and callogenesis and cell suspension cultures). In vitro systems were also used to study physiological, biochemical, and molecular processes in halophytes, such as functional and salt-tolerance studies. Thus, the application of PTC to halophytes may be used to improve their controlled multiplication and the selection of desired traits for the in vitro production of plants enriched in nutritional and functional components, as well as for the study of their resistance to salt stress.