Optimization of microwave-assisted extraction of carotenoids from Citrus clementina peels

Background: Citrus fruits, especially clementines, are among the most consumed fruits in the world. Clementine consists of pulp (endocarp) and peel (epicarp) which are rich in carotenoids. After using fruit pulp, peels are usually discarded as waste; the valorization of the latter in the recovery of its beneficial components, mainly carotenoids, may seem to be important. Objective: The main objective of this study is to determine the optimal conditions allowing the extraction of a high carotenoids yield from clementine peels. Methods: The microwave-assisted extraction method (MAE) was applied for extraction of total carotenoids from Citrus clementina peels, and the response surface methodology (RSM) was used to investigate the influence of extraction parameters, including hexane concentration, microwave power, irradiation time, and solvent to solid ratio, on the extraction yield, then the results were modeled using a second order regression. Total carotenoids yield of clementine peel extract obtained under optimal microwave-assisted extraction conditions was compared to extracts performed using two conventional extraction methods (maceration and Soxhlet). Results: The optimal conditions for microwave-assisted extraction were 68% of hexane concentration using 561 W of microwave power during 7.64 min of irradiation time, 43 ml/g of solvent-to-solid ratio using two successive extractions. Under optimized conditions of microwave-assisted extraction, the recovery of carotenoid content was 186.55 µg/g dry matter (DM), which was higher than that obtained by the two conventional methods, maceration extraction (ME; 160.53 µg/g DM) and Soxhlet extraction (SE; 162.68 µg/g DM). Conclusion: From this study, it can be concluded that microwave-assisted extraction is an efficient method for carotenoid recovery and considering its high yield in reduced time, it could be recommended for extraction of these bioactive compounds from clementine peels.

Analysis of CcGASA family members in Citrus clementina (Hort. ex Tan.) by a genome-wide approach

The Gibberellic Acid Stimulated Arabidopsis (GASA) proteins were investigated in the study to help understand their possible roles in fruit trees, particularly in Citrus. A total of 18 CcGASA proteins were identified and characterized in Citrus clementina via a genome-wide approach. It was shown that the CcGASA proteins structurally shared a conserved GASA domain but varied considerably in primary sequences and motif compositions. Thus, they could be classified into three major phylogenetic groups, G1~G3, and two groups, G1 and G3 could be further classified into subgroups. The cis- elements on all CcGASA promoters were identified and categorized, and the associated transcription factors were predicted. In addition, the possible interactions between the CcGASA proteins and other proteins were predicted. All the clues suggested that these genes should be involved in defense against biotic and abiotic stresses and in growth and development. The notion was further supported by gene expression analysis that showed these genes were more or less responsive to the treatments of plant hormones (GA3, SA, ABA and IAA), and infections of citrus canker pathogen Xanthomonas citri. It was noted that both the segmental and the tandem duplications had played a role in the expansion of the CcGASA gene family in Citrus. Our results showed that the members of the CcGASA gene family should have structurally and functionally diverged to different degrees, and hence, the representative group members should be individually investigated to dissect their specific roles.

THE EFFECT OF IRRIGATION DOSES ON THE PRODUCTIVITY OF Citrus clementina variety “Fina Berkane”, A VARIETY OF CLEMENTINE PRODUCED IN TRIFFA PLAIN, NORTH EASTERN MOROCCO

Morocco is a Mediterranean country with an economy based on exporting agricultural products mainly to Europe. However, the ongoing climate change characterized by the frequency and the intensity of drought, along with the regularity of the rainfall is undeniably affecting the productivity of major agricultural products in Morocco. Citriculture is one of the key pillars of agricultural products in Moroccan conditions and the productivity of citriculture is tightly related to water management. To reduce this dependency under climatic change, a pilot study, over three years, has been launched to assess the effect of water stress on the production of clementine (Citrus clementina 'Berkane Clementine'). This study was carried out on the clementine, variety "Fina Berkane" at the Triffa plain, Northeastern Morocco. This variety was grafted on Volkameria rootstock with a density of 3m x 6m. Experimental variety was planted during October 2012, in loam-clay soil equipped with a drip irrigation system. Four doses of water irrigation (60%, 80%, 100%, and the conventional irrigation dose⩾120%) were tested based on the real water demand of the studied plant. Throughout the three-year monitoring, the effect of used water amount on the various parameters including yield, fruit size, and fruit quality have been studied. Results of the study revealed that the treatment 100% ETc provided a high number of fruits, yield, and a good quality ratio. It was also well reported that adopting a dose of 80% ETc could save a significant amount of irrigation water (more than 40% ETc) compared to the conventional irrigation dose (⩾120% ETc) without significantly affecting either the yield or the desired caliber profile (marketable standard).

Preparation of Multifunctional N-Doped Carbon Quantum Dots from Citrus clementina Peel: Investigating Targeted Pharmacological Activities and the Potential Application for Fe3+ Sensing

Carbon quantum dots (CQDs) have recently emerged as innovative theranostic nanomaterials, enabling fast and effective diagnosis and treatment. In this study, a facile hydrothermal approach for N-doped biomass-derived CQDs preparation from Citrus clementina peel and amino acids glycine (Gly) and arginine (Arg) has been presented. The gradual increase in the N-dopant (amino acids) nitrogen content increased the quantum yield of synthesized CQDs. The prepared CQDs exhibited good biocompatibility, stability in aqueous, and high ionic strength media, similar optical properties, while differences were observed regarding the structural and chemical diversity, and biological and antioxidant activity. The antiproliferative effect of CQD@Gly against pancreatic cancer cell lines (CFPAC-1) was observed. At the same time, CQD@Arg has demonstrated the highest quantum yield and antioxidant activity by DPPH scavenging radical method of 81.39 ± 0.39% and has been further used for the ion sensing and cellular imaging of cancer cells. The obtained results have demonstrated selective response toward Fe3+ detection, with linear response ranging from 7.0 µmol dm−3 to 50.0 µmol dm−3 with R2 = 0.9931 and limit of detection (LOD) of 4.57 ± 0.27 µmol dm−3. This research could be a good example of sustainable biomass waste utilization with potential for biomedical analysis and ion sensing applications.

Influence of Rootstock Genotype and Ploidy Level on Common Clementine (Citrus clementina Hort. ex Tan) Tolerance to Nutrient Deficiency

Nutrient deficiency, in particular when this involves a major macronutrient (N, P, and K), is a limiting factor on the performance of plants in their natural habitat and agricultural environment. In the citrus industry, one of the eco-friendliest techniques for improving tolerance to biotic and abiotic stress is based on the grafting of a rootstock and a scion of economic interest. Scion tolerance may be improved by a tetraploid rootstock. The purpose of this study was to highlight if tolerance of a common clementine scion (C) (Citrus clementina Hort. ex Tan) to nutrient deficiency could be improved by several diploid (2×) and their tetraploid (4×) counterparts citrus genotypes commonly used as rootstocks: Trifoliate orange × Cleopatra mandarin (C/PMC2x and C/PMC4x), Carrizo citrange (C/CC2x and C/CC4x), Citrumelo 4475 (C/CM2x and C/CM4x). The allotetraploid FlhorAG1 (C/FL4x) was also included in the experimental design. The impact of nutrient deficiency on these seven scion/rootstock combinations was evaluated at root and leaf levels by investigating anatomical parameters, photosynthetic properties and oxidative and antioxidant metabolism. Nutrient deficiency affects foliar tissues, physiological parameters and oxidative metabolism in leaves and roots in different ways depending on the rootstock genotype and ploidy level. The best known nutrient deficiency-tolerant common clementine scions were grafted with the doubled diploid Citrumelo 4475 (C/CM4x) and the allotetraploid FlhorAG1 (C/FL4x). These combinations were found to have less foliar damage, fewer changes of photosynthetic processes [leaf net photosynthetic rate (Pnet), stomatal conductance (gs), transpiration (E), maximum quantum efficiency of PSII (Fv/Fm), electron transport rate (ETR), ETR/Pnet], and effective quantum yield of PSII [Y(II)], less malondialdehyde accumulation in leaves and better functional enzymatic and non-enzymatic antioxidant systems. Common clementine scions grafted on other 4× rootstocks did not show better tolerance than those grafted on their 2× counterparts. Chromosome doubling of rootstocks did not systematically improve the tolerance of the common clementine scion to nutrient deficiency.

Profile of Fork-Tailed Bush Katydid (Orthoptera: Tettigoniidae) Feeding on Fruit of Clementine Mandarins

Sweet oranges (Citrus sinensis (L.) Osbeck Sapindales: Rutaceae) dominated commercial citrus production in California until recently when there has been a shift to mandarins, mostly Citrus reticulata (Blanco) mandarins and Citrus clementina (hort. ex Tanaka) clementines. Past analyses of commercial field scouting and harvest data indicated that fork-tailed bush katydids (Scudderia furcata Brunner von Wattenwyl), a major pest in oranges, are present in clementine groves, but that fruit scarring attributed to katydids is rare. Conversely, jagged or web-like scarring attributed to caterpillars was more prevalent than expected. We used two field experiments in four representative cultivars of clementines to test four explanatory hypotheses for this observation: 1) katydids do not feed on clementine fruit, 2) damaged clementine fruit recover, 3) damaged clementine fruit preferentially abscise, and 4) katydid scars on clementine fruit have a different, undocumented morphology, not recognized as katydid damage. We find support for the latter two hypotheses. Katydids fed readily on the clementine fruit of all cultivars tested, chewing irregular holes that developed into jagged or web-like scars of a range of shapes and often led to splitting and abscission of maturing fruit. The katydid scars often more closely resembled chewing caterpillar damage than the round katydid scars in oranges, suggesting that katydid damage is being misclassified in clementines. The resistance documented in some other mandarins was not observed. Katydids are clearly a frugivorous pest causing previously unrecognized scarring in clementines.

Caractérisation chimique et évaluation de l’activité antimicrobienne des flavonoïdes extraits des feuilles de Citrus clementina de la région de la Mitidja (Algérie)

Les agrumes occupent une superficie importante de 65 000 ha en Algérie (2014), leurs fruits sont utilisés dans l’alimentation humaine sous forme fraîche ou transformée, mais les feuilles sont communément utilisées en phytothérapie traditionnelle. Les bienfaits constatés par la population nous ont incités à valoriser les feuilles de Citrus clementina récupérées après la taille des arbres, par l’évaluation de l’activité antimicrobienne et la caractérisation chimique de leurs extraits flavonoïdes. L’étude quantitative en composés flavonoïdes montre un rendement de 11,2 % en flavonesflavonols, de 12,36 % en anthocyanes, de 17,87 % en hétérosides et de 12,1 % en C-glycosides. L’étude par spectrophotomètre UV-visible révèle que l’extrait des feuilles renferme 2,48 ± 0,36 mg/g de flavones-flavonols, 3,33 ± 0,296 mg/g de C-glucosides, 0,34 ± 0,05 mg/g d’anthocyanes et 3,77 ± 0,20 mg/g d’hétérosides. L’analyse par HPLC de l’extrait de feuilles de clémentinier révèle sa contenance en acide gallique (35,849 %), acide ascorbique (35,849 %), acide caféique (area = 6,1803 %), acide benzoïque (6,1125 %), catéchine (23,4311 %) et rutine (6,0082 %). Ces extraits montrent une activité antimicrobienne variable à l’égard de tous les germes microbiens (Gram+, Gram– et levure) étudiés. L’extrait flavones-flavonols montre une forte activité antimicrobienne à l’égard des souches Staphylococcus aureus, Staphylococcus epdermidis, Echerichia coli, Pseudomonas aeruginosa et Schacharomyces cervisai avec un diamètre supérieur à 20 mm et une concentration minimale d’inhibition (CMI) égale à 382, 150, 382 μg/ml et 0,380 mg/ml respectivement. Les autres extraits (anthocyanes et C-glycosides) ont un effet intermédiaire, faible ou absent sur les micro-organismes testés. Le pouvoir bactériostatique (CMI) de Staphylococcus aureus en milieu liquide est de 238 μg/ml, et le pouvoir bactéricide (concentration minimale bactéricide) est de 1,845 μg/ml. Cette méthode de dilution est plus précise que celle de milieu gélosé.