Evaluation of Physicochemical, Functional and Sensory Properties of Carob Pulp Beverage (Ceratonia Siliqua L)

In Morocco, carob tree (Ceratonia siliqua L.) is of considerable socio-economic importance, helping to improve the income of rural populations and to develop the economy of mountain areas. However, the primary use of the carob’s pulp is generally limited in Morocco to animal feed. As a result, the current research suggests a novel pulp-based product designed for human consumption. The main purpose of this work is to characterize a new formulated carob beverage and assess its physicochemical, biochemical, and sensory quality. Two formulas of carob beverages were prepared with two different sugar concentrations (formula A:5 g/100 mL sugar; formula B: 10 g/100 mL sugar). Measurements of pH, titrable acidity, and colour were performed to assess the physicochemical parameters. Total polyphenols and tannins contents were determined as important functional and biochemical parameters. Regarding sensory analysis, preference test, triangle test, hedonic test, and descriptive test, were evaluated. The colour of the two formulations was dark red orange, as determined by chromameter method. For formulae A and B, the titrable acidity was (1.450±0.025) g Citric Acid Equivalent (CAE) /L and (1.450±0.031) CAE g/L, respectively, and the Brix was (11.67±0.22) °Bx and (15.33±0.22) °Bx, respectively. The polyphenols content was (98.8±0.0019) mg Gallic Acid Equivalent (GAE) /100 mL for formula A and (97.7000±0.0017) mg GAE/100 mL for formula B. The tannin level was (24.500± 0.027) mg Catechin (CE)/100 mL and (25.400±0.017) mg CE/100 mL for the formula A and formula B samples, respectively. The sensory analysis tests revealed a distinct difference between the two formulae. In fact, the formula B was highly preferred compared to formula A, also the most prominent features in the drink were colour, smell, flavour, and sweetness.

Genome-wide footprints in the carob tree (Ceratonia siliqua) unveil a new domestication pattern of fruit trees in the Mediterranean

Intense research efforts on phylogeography over the last two decades uncovered major biogeographical trends and renewed our understandings of plant domestication in the Mediterranean. We aim to investigate the evolutionary history and the origin of domestication of the carob tree that has been cultivated for millennia for food and fodder. We used >1000 microsatellite genotypes to identify carob evolutionary units (CEUs) based on genetic diversity structure and geography. We investigated genome-wide diversity and evolutionary patterns of the CEUs with 3557 SNPs generated by restriction-site associated DNA sequencing (RADseq). The 56 populations sampled across the Mediterranean basin, classified as natural, semi-natural or cultivated, were examined. Although, RADseq data are consistent with previous studies identifying a strong West-to-East genetic structure and considerable admixture in some geographic parts, we reconstructed a new phylogeographic scenario with two migration routes occurring from a single refugium likely located in South-Western Morocco. Our results do not favour the regionally bound or single origin of domestication. Indeed, our findings support a cultivation model of locally selected wild genotypes, albeit punctuated by long-distance westward dispersals of domesticated varieties by humans, concomitant with major cultural waves by Romans and Arabs in the regions of dispersal. Ex-situ efforts to preserve carob genetic resources should prioritize accessions from both western and eastern populations, with emphasis on the most differentiated CEUs situated in South-Western Morocco, South Spain and Eastern Mediterranean. Our study underscores the relevance of natural and seminatural habitats of Mediterranean forests and their refugia in the conservation efforts of tree crops.

Anti-Cancer Activity and Phenolic Content of Extracts Derived from Cypriot Carob (Ceratonia siliqua L.) Pods Using Different Solvents

Extracts derived from the Ceratonia siliqua L. (carob) tree have been widely studied for their ability to prevent many diseases mainly due to the presence of polyphenolic compounds. In this study, we explored, for the first time, the anti-cancer properties of Cypriot carobs. We produced extracts from ripe and unripe whole carobs, pulp and seeds using solvents with different polarities. We measured the ability of the extracts to inhibit proliferation and induce apoptosis in cancer and normal immortalized breast cells, using the MTT assay, cell cycle analysis and Western Blotting. The extracts’ total polyphenol content and anti-oxidant action was evaluated using the Folin–Ciocalteu method and the DPPH assay. Finally, we used LC-MS analysis to identify and quantify polyphenols in the most effective extracts. Our results demonstrate that the anti-proliferative capacity of carob extracts varied with the stage of carob maturity and the extraction solvent. The Diethyl-ether and Ethyl acetate extracts derived from the ripe whole fruit had high Myricetin content and also displayed specific activity against cancer cells. Their mechanism of action involved caspase-dependent and independent apoptosis. Our results indicate that extracts from Cypriot carobs may have potential uses in the development of nutritional supplements and pharmaceuticals.

Volatile Profile of Portuguese Monofloral Honeys: Significance in Botanical Origin Determination

The volatile profiles of 51 samples from 12 monofloral-labelled Portuguese honey types were assessed. Honeys of bell heather, carob tree, chestnut, eucalyptus, incense, lavender, orange, rape, raspberry, rosemary, sunflower and strawberry tree were collected from several regions from mainland Portugal and from the Azores Islands. When available, the corresponding flower volatiles were comparatively evaluated. Honey volatiles were isolated using two different extraction methods, solid-phase microextraction (SPME) and hydrodistillation (HD), with HD proving to be more effective in the number of volatiles extracted. Agglomerative cluster analysis of honey HD volatiles evidenced two main clusters, one of which had nine sub-clusters. Components grouped by biosynthetic pathway defined alkanes and fatty acids as dominant, namely n-nonadecane, n-heneicosane, n-tricosane and n-pentacosane and palmitic, linoleic and oleic acids. Oxygen-containing monoterpenes, such as cis- and trans-linalool oxide (furanoid), hotrienol and the apocarotenoid α-isophorone, were also present in lower amounts. Aromatic amino acid derivatives were also identified, namely benzene acetaldehyde and 3,4,5-trimethylphenol. Fully grown classification tree analysis allowed the identification of the most relevant volatiles for discriminating the different honey types. Twelve volatile compounds were enough to fully discriminate eleven honey types (92%) according to the botanical origin.

Effect of Sewage Sludge and Palm Fiber Co-compost as horticultural substrate on Carob tree (Ceratonia siliqua) growth

The palm (Phoenix dactylifera) is the main species of the main agro-systems of semi-arid and arid bioclimatic stages in Algeria. Its fibers (FP) are widely available and not used. In addition, the available sewage sludge (SS) requires recovery for ecological and economic purposes. To this end, the objective of this work was to formulate a Co-composting (CC) composed of SS and PF as a horticultural substrate (HS) intended for the breeding of the carob tree (Ceratonia siliqua). During the six months of CC, the indicative maturity pa-rameters such as pH, electro conductivity (EC), dissolved O2 and dry matter (DW) showed stability of the product. Four HS composed of 4 different CC-soil (TS) ratios: T (8-2), T (7-3), T (6-4) and T (0-10) were studied in the labor-atory. The best physicochemical characteristics such as total porosity (TP), humidity rate (RH), bulk density (BD), drainage rate (DR), pH and EC were obtained in T (80:20). This same SH also gave the best growth parameters of plants such as root volume (RV), root architecture (RA), number of roots (RN), root length (RL), number of leaves (LN), stem length (SL), fresh aerial and root weight (AFW), (RFW), dry aerial and root weight (ADW) and (RDW). We conclude that T (8: 2) induced the best growth performance of C. siliqua.

Bioclimatic Impact on the Carob Seeds Morphological Diversity in Morocco

The carob tree, Ceratonia siliqua L., a typical thermophilic sclerophyllous species, is a multipurpose tree (agronomic, silvicultural and pastoral) with aromatic and medicinal properties. Due to its geographical distribution, it occupies different countries of the Mediterranean. In Morocco, this species spreads over the different bioclimatic from the arid in the south to the humid in the north. The present work, study the morphological diversity of the seeds of five carob tree ecotypes distributed along a rainfall gradient (from the arid in the south to the humid one towards the north of the country). The shape parameters studied are the length, width, thickness, weight, volume and density of the seeds.The results obtained show that the carob tree ecotypes are characterized by significant differences in the different morphological parameters of the seeds among the five ecotypes studied. Indeed, dry land seeds are generally reduced in length, volume and weight, while those in humid regions are larger in weight and size. Those of the dry sowing areas have moderate parameters. The variance analysis shows that the origin of the ecotypes hada significant effect on the various shape parameters. Moreover, the multiple comparison of averages revealed different distinct groups. ACP analysis revealed that seed weight and volume is strongly correlated with the first component (the bioclimatic origin and the annual rainfall of the site of origin). While the second component corresponds to the parameters of length, width and thickness of the seeds. As for the factor 3, it corresponds to the density of the seeds which is not dependent on the other variables.In terms of conclusion, it is deduced that in Morocco, the carob tree seeds are characterized by a large morphological diversity which varies according to the rainfall gradient.

Carob tree micropropagation essays (Ceratonia siliqua L.)

Carob tree (Ceratonia siliqua L.) is an agro-sylvo-pastoral species that has been classified among the most efficient trees. The first publications on the "in-vitro" multiplication of Carob are very recent and show conflicting results. Mastery of this technique seems necessary to provide the increased demand of the international market for the gum extracted from the endosperm of the seed, a polyoside called galactomannan. This work aimed to study micropropagation from adult tree micro cuttings. Our results showed that the most favorable disinfectant is HgCl2 with 93% surface sterile explants. Comparison of the effect of various hormones with different concentrations (BAP, Zeatin, GA3, TDZ, and ANA) has shown that the addition of BAP on MS medium gave a good response of axillary bud development (58,3%±6,42) while BAP supplemented with activated charcoal gave the best results (87,3%±4,23). As for rooting, the different alternatives of hormones types and concentrations must be later considered to initiate rooting of the buds.

Determination of a sex-related RAPD marker in Carob (Ceratonia siliqua L.)

Molecular markers are used in the characterization and breeding of organisms. Carob (Ceratonia siliqua L.) is a species with both dioecious and hermaphrodite flower forms. The determination of sex at an early stage of growth in this species, whose juvenility period is long, is important in breeding studies. The objective of this study was to identify the sex-related markers using RAPD method. Ten genotypes were obtained from natural F1 hybrids between a naturally grown a female and a male carob tree. DNA was extracted from the leaves of 12 carob plants. Using BSA, the female and male bulks were formed from five female and five male plants, respectively, using equal amounts of DNA from each plant. In this study, 130 RAPD primers were tested. That of 21 primers tested showed polymorphisms between male and female bulks. While the fragment of 750 bp from the OPA17 RAPD primer was not detected in the female parent, female bulk, and female F1 hybrids; it was observed in the male parent and four out of five male F1 hybrids. This is the first report in the literature that one RAPD marker, namely OPA17-750, related to 80% reliability to male sex in carob was determined.