Browsing by Author "Romano, Anabela"
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- Acacia wood fractionation using deep eutectic solvents: extraction, recovery, and characterization of the different fractionsPublication . Magalhães, Solange; Moreira, Adriana; Almeida, Ricardo; Cruz, Pedro Fernandes; Alves, Luís; Costa, Carolina; Mendes, Cátia; Medronho, Bruno; Romano, Anabela; Carvalho, Maria da Graça; Gamelas, José A. F.; Rasteiro, Maria da GraçaThe selective extraction and recovery of different lignocellulosic molecules of interest from forestry residues is increasing every day not only to satisfy the needs of driving a society toward more sustainable approaches and materials (rethinking waste as a valuable resource) but also because lignocellulosic molecules have several applications. For this purpose, the development of new sustainable and ecologically benign extraction approaches has grown significantly. Deep eutectic solvents (DESs) appear as a promising alternative for the processing and manipulation of biomass. In the present study, a DES formed using choline chloride and levulinic acid (ChCl:LA) was studied to fractionate lignocellulosic residues of acacia wood (Acacia dealbata Link), an invasive species in Portugal. Different parameters, such as temperature and extraction time, were optimized to enhance the yield and purity of recovered cellulose and lignin fractions. DESs containing LA were found to be promising solvent systems, as the hydrogen bond donor was considered relevant in relation to lignin extraction and cellulose concentration. On the other hand, the increase in temperature and extraction time increases the amount of extracted material from biomass but affects the purity of lignin. The most promising DES system, ChCELA in a ratio of 1:3, was found to not significantly depolymerize the extracted lignin, which presented a similar molecular weight to a la-aft lignin. Additionally, the P-31 NMR results revealed that the extracted lignin has a high content of phenolic OH groups, which favor its reactivity. A mixture of ChCl:LA may be considered a fully renewable solvent, and the formed DES presents good potential to fractionate wood residues.
- Accumulation of phenolic compounds in in vitro cultures and wild plants of Lavandula viridis L'Her and their antioxidant and anti-cholinesterase potentialPublication . Costa, Patrícia; Gonçalves, Sandra; Valentão, Patrícia; Andrade, Paula B.; Romano, AnabelaIn this study, we evaluated the phenolic profile, antioxidant and anti-cholinesterase potential of different extracts from wild plants and in vitro cultures of Lavandula viridis L'Her. The HPLC-DAD analysis allowed the identification and quantification of 3-0-caffeoylquinic, 4-0-caffeoylquinic, 5-0-caffeoylquinic and rosmarinic acids, and luteolin and pinocembrin. Water/ethanol extract from in vitro cultures contained the highest amount of the identified phenolic compounds (51652.92 mg/kg). To investigate the antioxidant activity we used Trolox equivalent antioxidant capacity, oxygen radical absorbance capacity, Fe2+ chelation activity and the inhibition of Fe2+-induced lipid peroxidation in mouse brain homogenates (in vitro). Overall, all the extracts from both wild plants and in vitro cultures exhibited ability to scavenge free radicals, to chelate Fe2+ and to protect against lipid peroxidation. In addition, the extracts from L. viridis were active in inhibiting both acetylcholinesterase and butyrylcholinesterase (Ellman's method). Our findings suggest that L. viridis in vitro cultures represent a promising alternative for the production of active metabolites with antioxidant and anti-cholinesterase activity. (C) 2013 Elsevier Ltd. All rights reserved.
- Allelopathic effects of extracts from several plant species on seed germination and seedlings growth of Lactuca sativa LPublication . Gonçalves, Sandra; Franco, J.; Romano, AnabelaThe ability of a plant species to inhibit germination of other plants is an untapped resource for weed control in crops that could revolutionize organic crop production. The main objective of this study was to evaluate the allelopathic potential of aqueous (5 and 10%, w/v) and methanolic (50mg ml-1) extracts from several plant species (Arbustus unedo L., Ericaceae; Daphne gnidium L., Thymeleaceae; Myrtus communis L., Myrtaceae; Olea europea L., Oleaceae; Pistacia lentiscus L., Anacardiacceae; Quercus suber L., Fagaceae; Ruscus aculeatus L., Liliaceae, Rhododendron ponticum subsp baeticum (Boissier & Reuter) Handel-Mazzetti, Ericaceae) on seed germination and seedling growth of Lactuca sativa. To test the phytotoxicity 500µl of the extract were added separately over filter papers (9cm diameter) in Petri dishes. It was observed that methanolic extracts from M. communis and R. aculeatus completely inhibited seed germination, and almost all the extracts tested reduced seed germination and/or seedling growth. The inhibition of root growth was generally higher than of shoot growth, with the highest percentages of root inhibition being observed with the aqueous extracts of D. gnidium (80 and 72% for the extract at 5 and 10%, respectively) and R. aculeatus (74 and 78%), and with the methanolic extract of Q. suber (74%). Besides inhibiting seedling growth other morphological abnormalities occurred in the presence of the extracts. Roots of treated plants were thicker with a brownish color as compared to controls. Studies are being conducted to investigate the phytotoxic effect of the extracts on physiological and biochemical processes during germination and seedlings growth.
- Aluminium speciation and internal detoxification mechanisms in plants: where do we stand?Publication . Grevenstuk, Tomás; Romano, AnabelaAluminium (Al) typically has a deleterious impact on plant growth and development but some plants can accumulate otherwise toxic concentrations of this metal by deploying internal detoxification mechanisms (tolerance mechanisms). Although there are scattered reports concerning the regulation of Al uptake, transport and storage in metabolically innocuous forms, the overall picture remains incomplete and there are no accounts of a general overview of Al tolerance mechanisms. This review aims to compile the current knowledge on the physiological basis of Al tolerance and speculate on possible underlying molecular mechanisms regarding the uptake of Al, speciation in internal organs, transport to upper organs and storage in non-vital structures. Given the difficulties in detecting and studying the speciation of Al, special attention will be given to the approaches used so far and new developments that allow greater sensitivity and accuracy in uncovering the fate of Al in planta. © 2013 The Royal Society of Chemistry.
- Aluminum inhibits root growth and induces hydrogen peroxide accumulation in Plantago algarbiensis and P. almogravensis seedlingsPublication . Martins, Neusa; Gonçalves, Sandra; Romano, AnabelaWe have evaluated the impact of aluminum (Al) on germination, relative root growth, Al accumulation in roots tips, H2O2 levels, plasma membrane integrity, pigment levels, protein content, and the activities of superoxide dismutase (SOD) and catalase (CAT) in seedlings of the endangered Portuguese species Plantago algarbiensis and Plantago almogravensis. We found that up to 400 μM Al had no impact on the germination percentage in either species but inhibited root growth in a concentration-dependent manner (more severely in P. algarbiensis). Al accumulation in the root tips of both species was concentration dependent up to 200 μM but declined thereafter despite the absence of membrane damage. We observed a concentration-dependent induction of SOD activity but no change in CAT activity resulting in the accumulation of H2O2 (a known growth inhibitor), although its impact in P. almogravensis may be partially ameliorated by the accumulation of carotenoid pigments. Our data suggest an association between Al uptake, H2O2 production, and the inhibition of root growth during early seedling development in P. algarbiensis and P. almogravensis, although the latter is more tolerant towards higher concentrations of the metal. © 2013 Springer-Verlag Wien.
- An efficient in vitro propagation protocol for Pinguicula lusitanica, a rare insectivorous plantPublication . Gonçalves, Sandra; Escapa, Ana Luísa; Grevenstuk, Tomás; Romano, AnabelaIn this study, an efficient protocol was developed for in vitro propagation of Pinguicula lusitanica L., a rare insectivorous plant with pharmacological value and limited reproductive capacity. The effects of two concentrations (0.1 and 0.5 mg l-1) of a range of plant growth regulators, including cytokinins (BA, KIN, and ZEA) and auxins (IAA, IBA, and NAA), and three concentrations of MS medium macronutrients (total, 1/2 and 1/4MS) on proliferation and rooting, were investigated. P. lusitanica shoots showed abundant proliferation and rooting capacity, both of which were significantly influenced by MS medium strength. The use of 1/2MS supplemented with 0.5 mg l-1 BA or KIN ensure a 29-fold rate of proliferation. Best rooting frequency and higher root number and length were attained in 1/4MS medium containing 0.2 mg l-1 IAA. Sixty percent of the plantlets were successfully acclimatized to ex vitro conditions, exhibiting normal development. These plantlets can be used to replenish declining populations in the wild and/or for the extraction of bioactive compounds, reducing pressure on wild stocks.
- Analysis of the volatiles emitted by whole flowers and isolated flower organs of the carob tree using HS-SPME-GC/MSPublication . Custódio, Luísa; Serra, Hugo; Nogueira, José Manuel F.; Gonçalves, Sandra; Romano, AnabelaThe volatiles emitted by fresh whole flowers and isolated flower organs of male, female, and hermaphrodite carob trees (Ceratonia siliqua L.; Leguminosae)were analyzed by headspace solid-phase microextraction followed by capillary gas chromatography and mass spectrometry. The headspace of carob flowers is mainly constituted of high amounts of monoterpenes and sesquiterpenes, and more than 25 compounds were identified. The gender and cultivar affected both the qualitative profile and the relative abundances of the volatiles of whole flowers and isolated floral organs. Linalool and its derivatives (cis-linalool furan oxide, 2,2,6-trimethyl-3-keto-6-vinyltetrahydropyran, cis-linalool pyran oxide, and trans-linalool furan oxide), a-pinene, and a-farnesene were the dominant volatiles. Female flowers had a higher diversity of volatile compounds than males and hermaphrodites, but a lower abundance of the major ones. Similarly, the floral scent of female flowers of cv. Mulata had a higher content of volatiles but a lower abundance of the major ones, when compared to cv. Galhosa. In each of the three gender types of flowers, the nectary disks seemed to be the major source of volatiles.
- Antimicrobial activity of Drosophyllum lusitanicum, an endemic Mediterranean insectivorous plantPublication . Gonçalves, Sandra; Quintas, Célia; Gaspar, Maria Nelma; Nogueira, José Manuel F.; Romano, AnabelaThe aim of this study was to evaluate the antimicrobial activity of Drosophyllum lusitanicum leaf extract against various yeasts and bacteria species, including both standard and clinically isolated strains. The extract exhibited strong antimicrobial activity against all the tested yeast strains with inhibition zones ranging 23.67–42.23mm and with minimum inhibitory concentration (MIC) values ranging 31–63 mgL 1. All the Gram-positive bacteria studied were inhibited by the extract, showing inhibition zones ranging 17.67–43.00mm and MIC values comprising between 15.6 and 250 mgL 1. In contrast, the growth of the tested Gram-negative bacteria was not significantly affected by the extract. Among the microorganisms tested, Staphylococcus epidermidis ATCC 12228 was the most sensitive, presenting the lowest MIC value (15.6 mgL 1), while Enterococcus faecalis ATCC 29212 was the most tolerant (250 mgL 1). The extract of D. lusitanicum was analysed by gas chromatography-mass spectrometry and the major constituent found was plumbagin.
- Antioxidant activity and in vitro inhibition of tumor cell growth by leaf extracts from the carob tree (Ceratonia siliqua)Publication . Custódio, Luísa; Fernandes, Eliana; Escapa, Ana Luísa; López-Avilés, Sandra; Fajardo, Alba; Aligué, Rosa; Alberício, Fernando; Romano, AnabelaThe methanol leaf extracts of female cultivars of the carob tree [Ceratonia siliqua L. (Fabaceae)] and of hermaphrodite and male trees were investigated for their contents of phenolic compounds, their in vitro antioxidant activity, measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging and linoleic acid system assays, and their in vitro tumor growth inhibition on HeLa cells. The different cultivars and trees showed high levels of phenols, and considerable variations in the amount of these compounds. The extracts showed significant radical scavenging activity (RSA), which was not significantly affected by the gender of the tree. From the female cultivars tested, Galhosa exhibited the highest RSA. Gender significantly affected the antioxidant activity of the extracts measured by the linoleic acid system assay, and males and hermaphrodites showed the highest activities. The extracts displayed a remarkable ability to inhibit tumor cell proliferation, and their bioactivity varied with different cultivars or trees tested. Extracts from male and hermaphrodite trees exhibited higher capacity to inhibit the proliferation of HeLa cells than the female cultivars.
- Antioxidant activity and inhibitory effect of L. viridis extract on Fe2+-induced lipid peroxidation in brain homogenatesPublication . Costa, Patrícia; Gonçalves, Sandra; Romano, AnabelaThe brain is particularly susceptible to oxidative stress damaging effects due such events as the high consumption of oxygen, limited concentration of antioxidants and a relatively high degree of polyunsaturated fatty acids that are particularly good substrates for peroxidation reactions [1–3]. Oxidative stress could lead to damage biological target molecules, affecting the cellular function and integrity [4]. The ability of natural antioxidants, mainly phenolic compounds, to protect cells from oxidative stress has been previously demonstrated [5]. In this work, the methanol extract from Lavandula viridis L'Hér. (Lamiaceae), a xerophytic aromatic shrub endemic to the south-west Iberian Peninsula [6], was investigated for its effect on deoxyribose degradation, its reducing properties, Fe2+-chelating ability and total phenol content. The capacity of this extract to prevent Fe2+-induced lipid peroxidation in mouse brain (in vitro) was also evaluated. L. viridis extract showed Fe2+ chelating activity, reducing power and the ability to prevent Fe2+/H2O2-induced decomposition of deoxyribose in a dose-dependent manner. This extract also revealed a high phenol content (893.01±17.09µmol gallic acid equivalents/g extract) evaluated by Folin-Ciocalteu method. Moreover, in brain homogenates, the methanol extract of L. viridis caused a high decrease in the MDA production in both the basal and the Fe2+-induced lipid peroxidation. The effective protective properties of L. viridis could be attributed to its higher phenol content, Fe2+ chelating ability, reducing properties and HO· radical scavenging ability. The findings suggest that methanol extract from L. viridis could be a potential source of natural antioxidants.
- Antioxidant activity and verbascoside content in extracts from two uninvestigated endemic Plantago spp.Publication . Gonçalves, Sandra; Grevenstuk, Tomás; Martins, Neusa; Romano, AnabelaThis work describes for the first time the antioxidant activity of extracts from wild plants and in vitro cultures of two previously uninvestigated Plantago spp. endemic to Portugal, Plantago algarbiensis Samp. and Plantago almogravensis Franco. The Well-known and widely distributed species Plantago lagopus L. was also studied for comparison. Although the extracts from the three species exhibited radical scavenging and iron-reducing properties, the extracts from the two endemic species showed greater activity irrespectively of plant material source. Both P. algarbiensis extracts showed similar antioxidant activity whereas, in the case of P. almogravensis higher activity was observed for extracts from wild plants compared to extracts from in vitro cultures. Liquid chromatography-diode array detection analyses indicate verbascoside as a major constituent of all extracts and the highest content was obtained from in vitro cultures algarbiensis. In addition, a high correlation-between-the anti-oxidant-activity measured with three. methods and the total phenolic content was found, but no correlation was observed between antioxidant activity and verbascoside content. The results suggest that P. algarbiensis and P. almogravensis are sources of health-beneficial phytochemicals and indicate that in vitro cultures represent a promising alternative for the biosynthesis of verbascoside. (C) 2014 Elsevier B.V. All rights reserved.
- Antioxidant and anti-cholinesterase activities of Lavandula viridis L’Hér extracts after in vitro gastrointestinal digestionPublication . Costa, Patrícia; Grevenstuk, Tomás; Costa, Ana M. Rosa da; Gonçalves, Sandra; Romano, AnabelaLavandula viridis L’Hér is an aromatic shrub with relevant biological activities associated to the presence ofphenolic compounds. However, these compounds must be bioavailable to exert their biological function.Therefore, in this study, we investigated the influence of the L. viridis extract matrices on the digestion ofthe main component rosmarinic acid and their antioxidant and anti-cholinesterase activities after in vitrogastrointestinal digestion. Overall, the antioxidant and anti-cholinesterase activities were assured afterin vitro gastrointestinal processes and we observed that the L. viridis extract matrices have an importantrole in the bioactive effects of their main compound, rosmarinic acid. The L. viridis extracts and rosmarinicacid did not show any toxic effect on colon adenocarcinoma Caco-2 cell viability. Finally, the compoundsfrom L. viridis extracts were not metabolized by Caco-2 cells and were not able to permeate into them.
- Antioxidant and cytotoxic activities of carob tree fruit pulps are strongly influenced by Gender and cultivarPublication . Custódio, Luísa; Fernandes, Eliana; Escapa, Ana Luísa; Fajardo, Alba; Aligué, Rosa; Alberício, Fernando; Neng, Nuno Rosa; Nogueira, José Manuel F.; Romano, AnabelaExtracts from fruit pulps of six female cultivars and two hermaphrodite Portuguese carob trees [(Ceratonia siliqua L., Fabaceae)] exhibited strong antioxidant activity and were rich in phenolic compounds. The extracts decreased the viability of different human cancer cell lines on a dose- and time-dependent manner. Gender and cultivar significantly influenced the chemical content and the biological activities of the extracts. Extracts from hermaphrodite trees had a higher content of phenolic compounds, and exhibited higher antioxidant and cytotoxic activities. Among. females, cv. Aida had the highest radical scavenging activity and total content of phenolics, Mulata the highest capacity to inhibit lipid oxidation and Gasparinha the strongest cytotoxic activity on HeLa cells. The decrease in cell viability was associated with apoptosis on HeLa and MDA-MB-231 lines. (+)-Catechin and gallic acid (GA) were the main compounds identified in the extracts, and GA contributed to the antioxidant activity. Our results show that the antioxidant and cytotoxic activities of carob tree fruit pulps are strongly influenced by gender and cultivar, and provide new knowledge about the advantages of hermaphrodite trees over female cultivars, namely, as a source of compounds with biological interest, which may represent an increase of their agronomic interest.
- Antioxidant, cytotoxic and apoptotic activity of Drosophyllum lusitanicum extractsPublication . Gonçalves, Sandra; Xavier, Celine; Costa, Patrícia; Alberício, Fernando; Romano, AnabelaThis study was designed to determine the antioxidant, anti-proliferative, cytotoxic and apoptogenic properties of aqueous, methanol and hexane extracts from Drosophyllum lusitanicum leaves. Assays showed that the methanol extract had the highest antioxidant activity (F-C: 1188.06 ± 52.96 μmolGAE/mgextract; TEAC: 432.23 ± 6.71 μmolTE/mgextract; ORAC: 764.18 ± 61.18 μmolTE/mgextract)and the hexane extract had the lowest (F-C: 20.16 ± 1.62 μmolGAE/mgextract; TEAC: 20.94 ± 1.73 μmolTE/mgextract; ORAC: 50.66 ± 0.91 μmolTE/mgextract). The effect of plant extracts on HeLa cells proliferation and viability was evaluated by the crystal violet and WST-1 assays, respectively and apoptotic cells were determined using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). It was observed that the methanol extract inhibited cell proliferation and induced apoptosis on human cervical adenocarcinoma HeLa cells. Moreover, results indicated that the D. lusitanicum hexane extract exhibited cytotoxic effects on HeLa cells via apoptosis and also arrested cell cycle progression in G2/M phase. As damaging events are frequently correlated with oxidative stress, the prevalence of both antioxidant and anti-proliferative properties in the same plant species could be beneficial in terms of rational, preventive or therapeutic purposes. Thus D. lusitanicum may be a potential candidate in the field of anticancer drug discovery.
- Approaches for the cryopreservation of Plantago algarbiensis, a rare endemic species of the AlgarvePublication . Coelho, Natacha; González-Benito, María Elena; Romano, AnabelaPlantago algarbiensis is an endangered endemic species from the Algarve, Portugal. The main goal of this study was to investigate the viability of cryopreservation procedures in the conservation of seeds and nodal explants from this species. Seeds were directly immersed in liquid nitrogen (LN) for 30 days. Two methods were tested for the cryopreservation of nodal explants, namely droplet-vitrification and encapsulation-dehydration. For both methods, nodal segments were precultured on Murashige and Skoog (MS) medium and recovered on MS supplemented with 0.2 mg l(-1) 6-benzyladenine (BA), after freezing. After 30 days in LN, the germination capacity of seeds was not affected. The regrowth percentages of cryopreserved nodal segments were approximately 60%. With the droplet-vitrification method, a regrowth percentage of 60.0+/-15.2% was obtained after 120 min exposure to PVS2 (plant vitrification solution 2) and with encapsulation-dehydration method the highest percentage, 63.3+/-9.6%, was achieved after 3 h desiccation. Seed cryopreservation and cryopreservation of nodal segments by droplet-vitrification and encapsulation-dehydration are therefore effective approaches for the conservation of P. algarbiensis.
- Brief overview on bio-based adhesives and sealantsPublication . Magalhães, Solange; Alves, Luís; Medronho, Bruno; Fonseca, Ana C.; Romano, Anabela; Coelho, Jorge F.J.; Norgren, MagnusAdhesives and sealants (AS) are materials with excellent properties, versatility, and simple curing mechanisms, being widely used in different areas ranging from the construction to the medical sectors. Due to the fast-growing demand for petroleum-based products and the consequent negative environmental impact, there is an increasing need to develop novel and more sustainable sources to obtain raw materials (monomers). This reality is particularly relevant for AS industries, which are generally dependent on non-sustainable fossil raw materials. In this respect, biopolymers, such as cellulose, starch, lignin, or proteins, emerge as important alternatives. Nevertheless, substantial improvements and developments are still required in order to simplify the synthetic routes, as well as to improve the biopolymer stability and performance of these new bio-based AS formulations. This environmentally friendly strategy will hopefully lead to the future partial or even total replacement of non-renewable petroleum-based feedstock. In this brief overview, the general features of typical AS are reviewed and critically discussed regarding their drawbacks and advantages. Moreover, the challenges faced by novel and more ecological alternatives, in particular lignocellulose-based solutions, are highlighted.
- Cactus: chemical, nutraceutical composition and potential bio-pharmacological propertiesPublication . Das, Gitishree; Lim, Kyung Jik; Tantengco, Ourlad Alzeus G.; Carag, Harold M.; Goncalves, Sandra; Romano, Anabela; Das, Swagat Kumar; Coy-Barrera, Ericsson; Shin, Han-Seung; Gutierrez-Grijalva, Erick Paul; Heredia, J. Basilio; Patra, Jayanta KumarCactus species are plants that grow in the arid and semiarid regions of the world. They have long fascinated the attention of the scientific community due to their unusual biology. Cactus species are used for a variety of purposes, such as food, fodder, ornamental, and as medicinal plants. In the last regard, they have been used in traditional medicine for eras by the ancient people to cure several diseases. Recent scientific investigations suggest that cactus materials may be used as a source of naturally-occurring products, such as mucilage, fiber, pigments, and antioxidants. For this reason, numerous species under this family are becoming endangered and extinct. This review provides an overview of the habitat, classification, phytochemistry, chemical constituents, extraction and isolation of bioactive compounds, nutritional and pharmacological potential with pre-clinical and clinical studies of different Cactus species. Furthermore, conservation strategies for the ornamental and endangered species have also been discussed.
- Cardiovascular protective effect of cinnamon and its major bioactive constituents: An updatePublication . Das, Gitishree; Gonçalves, Sandra; Basilio Heredia, J.; Romano, Anabela; Jiménez-Ortega, Luis Alfonso; Gutiérrez-Grijalva, Erick P.; Shin, Han Seung; Patra, Jayanta KumarCinnamon from the bark of Cinnamomum species is one of the most important spices used worldwide in food and as a traditional medicine for centuries. It has substantial benefits for human health including its protective role on cardiovascular diseases. This review provides an overview of the cardiovascular protective effects of cinna-mon and its major bioactive constituents. Reviewed literature showed sufficient evidence that cinnamon can reduce the risk of cardiovascular diseases, including cardiac ischemia, cardiac hypertrophy, and myocardial infarction. Furthermore, cinnamon exhibited beneficial effects on cardiovascular-related comorbidities like diabetes, and other metabolic disorders, and showed antioxidant and anti-inflammatory effects. Cinnamon contains several bioactive compounds such as phenolics and volatile compounds. Cinnamaldehyde and cinnamic acid are among the main cinnamon compounds with protective effects on cardiovascular diseases through different molecular mechanisms. Although the protective effects of cinnamon and its main compounds have been extensively reported, more preclinical and clinical studies are still required before its use as a biopharmaceutical agent.
- Carob pulp: A nutritional and functional by-product worldwide spread in the formulation of different food products and beverages: A reviewPublication . Rodríguez-Solana, Raquel; Romano, Anabela; Moreno-Rojas, José ManuelCarob (Ceratonia siliqua L.) pod is a characteristic fruit from the Mediterranean regions. It is composed by seeds, the valuable part due to the extraction of locust bean gum, and the pulp, considered a by-product of the fruit processing industry. Carob pulp is a mixture of macro- and micronutrients, such as carbohydrates, vitamins and minerals, and secondary metabolites with functional properties. In the last few years, numerous studies on the chemical and biological characteristics of the pulp have been performed to encourage its commercial use. Its potential applications as a nutraceutical ingredient in many recipes for food and beverage elaborations have been extensively evaluated. Another aspect highlighted in this work is the use of alternative processes or conditions to mitigate furanic production, recognized for its toxicity. Furthermore, carob pulp’s similar sensorial, chemical and biological properties to cocoa, the absence of the stimulating alkaloids theobromine and caffeine, as well as its low-fat content, make it a healthier potential substitute for cocoa. This paper reviews the nutritional and functional values of carob pulp-based products in order to provide information on the proclaimed health-promoting properties of this interesting by-product
- Carob trees (Ceratonia siliqua L.) regenerated in vitro can acclimatize successfully to match the field performance of seed-derived plantsPublication . Osório, Maria Leonor; Osório, Júlio; Gonçalves, Sandra; David, Maria Manuela; Romano, Anabela; Correia, Maria JoãoThe use of in vitro regenerated plants in forestry and orchard depends ultimately on the development of efficient transplantation protocols, ensuring high survival rates and successful establishment under field conditions. We tested the performance of micropropagated carob trees (Ceratonia siliqua L.) throughout the acclimatization process in terms of survival, growth and physiological traits, including field comparisons with seed-derived and mother plants. The field trial was 100 %successful, i.e. we found no major differences between micropropagated, seed-derived and mother plants in terms of growth rate, height, number of leaves, photosynthetic efficiency, chlorophyll fluorescence, chlorophyll content and soluble protein content, although these parameters changed significantly during acclimatization. Stomatal conductance (gs) was reduced by fourfold when plants were transferred from in vitro culture to the growth chamber, thus preventing uncontrolled wilting. The photosynthetic rate (PN) was relatively low in vitro, in the growth chamber and the greenhouse, but increased to match seed-derived and mother plants in the field. The chlorophyll a/b ratio in leaves from in vitro and growth chamber plants was typical of shade plants (2.1) but became more characteristic of sun plants in the subsequent acclimatization stages (3.1–3.5). The maximum efficiency of photosystem II (Fv/Fm) remained mostly constant at ~0.80 throughout acclimatization, as is typical for healthy, non-stressed plants. We conclude that our micropropagation and acclimatization protocols provide a suitable alternative to traditional mass propagation techniques for the carob tree.