Influência do metil-jasmonato na formação de compostos voláteis do aroma em Bananas (Musa acuminata cv. Nanicão) e Laranjas (Citrus sinensis cv. Pera) / Influence of methyljasmonate on the formation of aroma volatile compounds in Bananas (Musa acuminata cv. Nanicão) and Oranges (Citrus sinensis cv. Pera)

O principal hormônio associado aos processos do amadurecimento é o etileno, porém, na formação de compostos voláteis nos frutos, observa-se que as auxinas, o ácido abscísico e os jasmonatos também podem atuar como reguladores. Estudos indicam que em frutos climatéricos deve haver uma interação entre o metil jasmonato (MeJA) e o etileno na formação de compostos voláteis, mas em frutos não-climatéricos tal interação não é tão evidente. Há evidências de que o MeJA atue na regulação de algumas vias metabólicas relacionadas ao amadurecimento em frutos, sendo capaz de induzir aumento na produção de várias classes de compostos voláteis, através da expressão de genes que codificam as enzimas relacionadas às suas vias biossintéticas. Neste sentido, o objetivo deste projeto foi avaliar o efeito do metil jasmonato sobre o padrão de produção de compostos voláteis do aroma em frutos climatéricos e não-climatéricos. Precedentes do laboratório de Química, Bioq. e Biol. Molecular de Alimentos indicam que o MeJA apresentou padrões diferentes de comportamento em frutos climatéricos e não-climatéricos no que tange a formação do aroma. Assim, o presente projeto tem por hipótese a diferença de influência que o MeJA exerce sobre a produção de compostos voláteis em frutos climatéricos e não-climatéricos. Para testar esta hipótese foi avaliado o efeito do tratamento com MeJA na produção de compostos voláteis do aroma durante o amadurecimento de banana (Musa acuminata, cv. Nanicão), como exemplo de fruto climatérico e laranja (Citrus sinensis cv Pêra) para não-climatéricos. Os frutos foram divididos em grupo controle e tratado com MeJA (10 ppm/24h), armazenados em caixas plásticas tampadas e lacradas. Após tratamento foram submetidos a análises diárias da produção de etileno por cromatrogafia gasosa (CG), cor da casca e pesagem. Baseado em escalas de cor e a polpa foi congelada em N2 líq. e armazenada a -80°C para posterior análise dos compostos voláteis por cromatografia gasosa acoplada à espectrometria de massas (GC-MS). Ésteres, álcoois, cetonas e aldeídos foram compostos majoritariamente identificados na banana e terpenos, aldeídos, ésteres na laranja. As Bananas sofreram influência no perfil de acetato de isoamila, butonoato de butila, isobutirato de isoamila e isolvalerato de isoamila do começo ao fim do tratamento com MeJA, e as laranjas o tratamento influenciou os compostos Cis-muirola-3-5-diene, gamageraniol, alfa-copaeno, valenceno, alfa-pineno, carvone, geranial, entre outros terpenos, aldeídos como 3-hexanal e 2-hexenal (E) e ésteres como butirato de etila, nerol e tiglato de etilo. Os ésteres em frutos são produzidos por várias isoformas das álcool acil transferases (AATs). Estudos explicam que, ao menos 31 transcritos de AATs foram identificados em bananas, sendo 8 com altos níveis de expressão. Assim, é plausível supor que tal variedade de transcritos, e por conseguinte as AATs que codificam, sejam reguladas por múltiplos fatores, o que pode incluir o MeJa dentre outros sinais hormonais. Os terpenos são formados a partir de duas rotas, a do ácido mevalônico (MVA) e a rota do metileritritol fosfato (MEP). Compostos como, D-limoneno (51) e beta-selineno (62) tiveram níveis relativos maiores nos frutos do grupo controle, enquanto compostos terpênicos como geranial (59), valenceno (79) e o-cimeno (128), apresentaram maiores níveis nos frutos tratados com MeJa, no primeiro dia após o tratamento. Os resultados mostraram que o tratamento hormonal com MeJA causou mudanças do início ao fim do amadurecimento na composição do aroma de bananas (Musa acuminata cv Nanicão) e laranjas (Citrus sinensis cv Pera)

The main hormone associated with ripening processes is ethylene, but in the formation of volatile compounds in fruits, auxins, abscisic acid and jasmonates can also act as regulators. Studies indicate that in climacteric fruits there should be an interaction between methyl jasmonate (MeJA) and ethylene in the formation of volatile compounds, but in nonclimacteric fruits such interaction is not so evident. There is evidence that MeJA acts in the regulation of some metabolic pathways related to fruit ripening, being able to induce an increase in the production of several classes of volatile compounds, through the expression of genes that encode enzymes related to their biosynthetic pathways. In this sense, the objective of this project was to evaluate the effect of methyl jasmonate on the production pattern of aroma volatile compounds in climacteric and non-climacteric fruits. Precedents from the Laboratory of Food Chemistry, Bioq. and Molecular Biol. Molecular Chemistry, Bioq. and Molecular Biol. of Foods laboratory indicate that MeJA showed different behavior patterns in climacteric and non-climacteric fruits regarding aroma formation. Thus, the present project hypothesizes the different influence that MeJA has on the production of volatile compounds in climacteric and non-climacteric fruits. To test this hypothesis the effect of MeJA treatment on the production of volatile aroma compounds during ripening of banana (Musa acuminata, cv. Nanicão) as an example for climacteric fruit and orange (Citrus sinensis cv Pêra) for non-climacteric fruit was evaluated. Fruits were divided into control and MeJA treated group (10 ppm/24h), stored in capped and sealed plastic boxes. After treatment they were subjected to daily analysis of ethylene production by gas chromatography (GC), peel color and weighing. Based on color scales and the pulp was frozen in liquid N2 and stored at -80°C for subsequent analysis of volatile compounds by gas chromatography coupled to mass spectrometry (GC-MS). Esters, alcohols, ketones and aldehydes were compounds mostly identified in banana and terpenes, aldehydes, esters in orange. Bananas were influenced in the profile of isoamyl acetate, butyl butonoate, isoamyl isobutyrate and isoamyl isolvalerate from the beginning to the end of the MeJA treatment, and oranges the treatment influenced the compounds Cis-myrola-3-5-diene, gamma-geraniol, alpha-copaene, valencene, alpha-pinene, carvone, geranial, among other terpenes, aldehydes like 3-hexanal and 2-hexenal (E), and esters like ethyl butyrate, nerol, and ethyl tiglate. Esters in fruits are produced by various isoforms of the alcohol acyl transferases (AATs). Studies explain that at least 31 AAT transcripts have been identified in bananas, 8 of which have high expression levels. Thus, it is plausible to assume that such a variety of transcripts, and therefore the AATs they encode, are regulated by multiple factors, which may include MeJa among other hormonal signals. Terpenes are formed from two routes, the mevalonic acid (MVA) route and the methylerythritol phosphate (MEP) route. Compounds such as, D-limonene (51) and beta-selinene (62) had higher relative levels in the fruits of the control group, while terpenic compounds such as geranial (59), valencene (79) and o-cymene (128), showed higher levels in the MeJa treated fruits on the first day after treatment. The results showed that the hormonal treatment with MeJA caused changes from the beginning to the end of ripening in the aroma composition of bananas (Musa acuminata cv Nanicão) and oranges (Citrus sinensis cv Pera)

Curcumin solid dispersion based on three model acrylic polymers: formulation and release properties

Abstract To investigate structure-property relationship of polymer-based curcumin solid dispersion (SD), three acrylic polymers were used to formulate curcumin SD by solvent evaporation method. Curcumin Eudragit EPO SD (cur@EPO), curcumin Eudragit RS PO SD (cur@RSPO) and curcumin Eudragit RL PO SD (cur@RLPO) showed deep red, golden orange and reddish orange color, respectively. Cur@RSPO entrapped 15.42 wt% of curcumin followed by cur@RL PO and cur@EPO. FTIR spectra indicated that in cur@EPO, curcumin may transfer hydrogen to the dimethylaminoethyl methacrylate group and thus change its color to red. In contrast, curcumin may form hydrogen bonding with Eudragit RS PO and Eudragit RL. Curcumin exists in amorphous state in three SDs as proved by differential scanning calorimetry and X-Ray diffraction measurement. In vitro digestion presented that lower pH value in simulated gastric fluid (SGF) stimulates the curcumin release from cur@EPO while permeability influences the release profile in other two SDs. When in simulated intestinal fluid (SIF), first order release model governs the release behaviors of all three SDs which showed sustained release pattern. Our results are helpful to elucidate how structure of polymer may impact on the major properties of curcumin contained SD and will be promising to broaden its therapeutic applications.

Efeitos da ingestão do suco de laranja Pera e suco de laranja Moro na microbiota intestinal e marcadores inflamatórios em indivíduos com obesidade / Effect of Pera and Moro orange juice consumption on the gut microbiota and inflammatory parameters in volunteers with obesity

A obesidade é uma doença complexa que está associada inflamação crônica de baixo grau que contribui para o desenvolvimento de diversos distúrbios metabólicos como a resistência à insulina e estudos recentes sugerem a influência da microbiota intestinal no desenvolvimento e manutenção da doença. Diversos estudos apontam para o benefício da ingestão de frutas e vegetais na prevenção e tratamento de doenças crônicas. O suco de laranja contém diversos compostos bioativos com ações anti-inflamatórias, antioxidantes com efeitos na composição da microbiota intestinal. Deste modo, o objetivo principal deste estudo foi avaliar os efeitos da ingestão do suco de laranja Pera e Moro sobre a composição da microbiota intestinal e de parâmetros inflamatórios em voluntários com obesidade e resistência à insulina. Foi realizado um ensaio clínico crossover com suplementação de suco de laranja (400ml/dia) por 15 dias com um período de washout de 40 dias. As análises de sangue, fezes, urina, composição corporal, consumo alimentar foram realizadas antes e após cada intervenção. A comparação entre os tratamentos foi realizada utilizando equações de estimativas generalizadas e adotou-se um nível de significância de 5%. Em relação à microbiota intestinal, em ambos os tratamentos, os dois filos mais abundantes foram Firmicutes e Actinobateria. Dos gêneros analisados, observou-se maior abundância de Bifidobacterium após a suplementação com o suco de laranja Moro. O suco de laranja Pera promoveu uma diminuição da zonulina e o suco de laranja Moro contribuiu para redução de citocinas inflamatórias, diminuição da pressão arterial e aumento nos níveis de acetato nas fezes. Após a separação dos voluntários por grau de obesidade, observamos que o suco de laranja Moro contribuiu para o aumento na abundância de Akkermansia, Alistipes, Bacteroides e Catenibacterium em indivíduos com obesidade grau 3. Além disso, em ambos os sucos encontramos redução da razão Firmicutes/Bacteroidetes e aumento da excreção de metabólitos de flavonoides após os tratamentos. Diante destes resultados, conclui-se que o suco de laranja Pera apresentou ações positivas sobre a permeabilidade intestinal e o suco de laranja Moro promoveu efeitos mais expressivos na modulação da inflamação associada à obesidade e da microbiota intestinal

Obesity is a complex disease that is associated with low-grade chronic inflammation, and it contributes to the development of several metabolic disorders such as insulin resistance, and recent studies suggest the influence of the intestinal microbiota in the development and maintenance of the disease. Several studies have suggested the benefit of fruits and vegetables consumption in the prevention and treatment of chronic diseases. The orange juice contains some bioactive compounds with anti-inflammatory and antioxidant actions with effects in the composition of the gut microbiota. Thus, the main objective of this study was to evaluate the effects of Pera and Moro orange juice consumption on the composition of the gut microbiota and inflammatory parameters in volunteers with obesity and insulin resistance. A crossover clinical trial was carried out with orange juice supplementation (400ml/day) for 15 days with a washout period of 40 days. Blood, feces, urine, body composition, food consumption were analyzed before and after each intervention. Comparison between treatments was performed using generalized estimating equations and a significance level of 5% was adopted. In relation to gut microbiota, in both treatments, the two most abundant phyla were Firmicutes and Actinobateria. In the analysis of bacterial genera, a greater abundance of Bifidobacterium was observed after supplementation with Moro orange juice. The Pera orange juice reduced zonulin and Moro orange juice contributed to a reduction on inflammatory cytokines, a decrease in blood pressure and an increase in acetate levels in the stool. After separating the volunteers by degree of obesity, we observed that Moro orange juice contributed to the increase in the abundance of Akkermansia, Alistipes, Bacteroides and Catenibacterium in individuals with grade 3 obesity. Furthermore, in both juices we found a reduction in the Firmicutes/Bacteroidetes ratio and increased excretion of flavonoid metabolites after treatments. Therefore, we concluded that Pera orange juice had positive actions on intestinal permeability and Moro orange juice promoted more expressive effects on the modulation of inflammation associated with obesity and on the intestinal microbiota

Comprehensive Analysis of Autophagy-Related Genes in Sweet Orange (Citrus sinensis) Highlights Their Roles in Response to Abiotic Stresses.

Autophagy is a highly conserved intracellular degradation pathway that breaks down damaged macromolecules and/or organelles. It is involved in plant development and senescence, as well as in biotic and abiotic stresses. However, the autophagy process and related genes are largely unknown in citrus. In this study, we identified 35 autophagy-related genes (CsATGs-autophagy-related genes (ATGs) of Citrus sinensis, Cs) in a genome-wide manner from sweet orange (Citrus sinensis). Bioinformatic analysis showed that these CsATGs were highly similar to Arabidopsis ATGs in both sequence and phylogeny. All the CsATGs were randomly distributed on nine known (28 genes) and one unknown (7 genes) chromosomes. Ten CsATGs were predicted to be segmental duplications. Expression patterns suggested that most of the CsATG were significantly up- or down-regulated in response to drought; cold; heat; salt; mannitol; and excess manganese, copper, and cadmium stresses. In addition, two ATG18 members, CsATG18a and CsATG18b, were cloned from sweet orange and ectopically expressed in Arabidopsis. The CsATG18a and CsATG18b transgenic plants showed enhanced tolerance to osmotic stress, salt, as well as drought (CsATG18a) or cold (CsATG18b), compared to wild-type plants. These results highlight the essential roles of CsATG genes in abiotic stresses.

Carotenoid biosynthesis and quality characteristics of new hybrids between tangor (Citrus reticulata x C. sinensis) cv. 'Murcott' and sweet orange (C. sinensis) cv. 'Pêra'.

Phenotypic characteristics, as well as the relation between carotenoid accumulation and gene expression during ripening were determined in fruits of five new hybrids between tangor cv. 'Murcott' and sweet orange cv. 'Pêra'. The genotypes were classified into the orange-like group, showing mainly epoxycarotenoids, oval fruit shape and yellowish color, or in the mandarin-like group, showing mainly ß-cryptoxanthin, flattened shape and deep-orange coloration; although some hybrids presented intermediate characteristics. The diversity in carotenoid composition of hybrids and genitors were mostly explained by patterns of gene expression. High carotenoid (250-426 µg/g dry weight [dw]) and ß-cryptoxanthin (81-125 µg/g dw) contents, observed in the mandarin-like group, were generally associated with high expression of upstream genes (GGPPS1, PSY, PDS). On the other hand, low expression/repression of these genes and high expression of downstream genes (BCHX and ZEP) were associated with low carotenoid (~158 µg/g dw) and ß-cryptoxanthin (5-22 µg/g dw) contents and epoxycarotenoid accumulation, as occurred in the orange-like group. Breeding experiments resulted in hybrids with outstanding higher carotenoid contents than both genitors (up to 426 µg/g dw versus 158-250 µg/g dw in genitors), which was attributed to transgressive segregation. Differences among genotypes have great impact on commercial fruit quality and potential health benefits, such as the provitamin A content.

Analysis of phytochemical contributors to antioxidant capacity of the peel of Chinese mandarin and orange varieties.

The phytochemicals in the peel of six oranges and ten mandarins including seven wild varieties and three cultivars were systematically characterised using UHPLC-Q-TOF-MS, and the correlation analysis was performed between phytochemicals and antioxidant capacity in order to investigate the phytochemical contributors to antioxidant capacity. The gradient elution was completed within 16 min and 92 compounds were undoubtedly or tentatively identified. Furthermore, the antioxidant capacities were determined using ABTS, DPPH and FRAP methods. The number of compounds, their contents and the antioxidant capacities were sequenced in the same order of the wild mandarins > cultivated mandarins > oranges. The correlation analysis that showed five compounds were significantly correlated with the antioxidant capacity and can act as main contributors to the citrus varieties with high antioxidant capacities. This study is systematic for the metabolites identification of mandarins and oranges and provides valuable information for effective utilisation of citrus peel and their bioactive compounds.

Daily Consumption of Orange Juice from Citrus sinensis L. Osbeck cv. Cara Cara and cv. Bahia Differently Affects Gut Microbiota Profiling as Unveiled by an Integrated Meta-Omics Approach.

We have investigated the effect of intake of two different orange juices from Citrus sinensis cv. "Cara Cara" and cv. "Bahia" on faecal microbiota and metabolome using an integrated meta-omics approach. Following a randomized crossover design, healthy subjects daily consumed 500 mL of orange juice from Cara Cara or Bahia juices or an isocaloric control drink. Stools were collected at baseline (T0) and after a week (T7) of intervention. Operational taxonomic units (OTUs) were pyrosequenced targeting 16S rRNA, and faecal metabolites were analyzed by an untargeted metabolomics approach based on 1H NMR spectroscopy. The major shift observed in microbiota composition after orange juice intake was the increased abundance of a network of Clostridia OTUs from Mogibacteriaceae, Tissierellaceae, Veillonellaceae, Odoribacteraceae, and Ruminococcaceae families, whose members were differently affected by Cara Cara or Bahia juice consumption. A core of six metabolites such as inositol, choline, lysine, arginine, urocanic acid, and formate significantly increased in Cara Cara compared to the Bahia group.

Phenolic composition and total antioxidant capacity of South African frozen concentrated orange juice as affected by varietal, seasonal and regional differences.

BACKGROUND: There is epidemiological evidence that fruits and vegetables promote general health due to their phenolic composition. The phenolic composition of three commercially important citrus varieties ('Mandarin', 'Navel' orange and 'Valencia' orange), used for frozen concentrated orange juice (FCOJ) production in South Africa, were evaluated based on variety, production season and geographical region (Western Cape (WC) and Eastern Cape (EC)). RESULTS: FCOJ from the WC had significantly (P < 0.05) lower titratable acidity (TA) and higher total soluble solids (TSS):TA ratio compared to FCOJ produced in the EC. The 'Navel' FCOJ, irrespective of season and region, had the highest (P < 0.05) phenolic content (sum of quantified compounds, TP). Regional effects were clear for the 'Mandarin' variety, the EC 'Mandarin' FCOJ had the highest TP and WC 'Mandarin' had the lowest. Seasonal differences were less evident. Variation that could be ascribed to regional differences were also found for individual phenolic compounds. CONCLUSION: Robust data regarding the phenolic profile of FCOJ produced in South Africa, suitable for inclusion in food composition databases, were collected. © 2018 Society of Chemical Industry.

Influence of paternal consumption of a high-fat/high-sugar diet and/or orange juice during preconception on C57BL/6 female mice offspring breast cancer risk / Influência do consumo paterno de uma dieta hiperlipídica/hiperglicídica e/ou suco de laranja durante a preconcepção no risco de câncer de mama na prole feminina de camundongos C57BL/6

Breast cancer is the most frequent cancer in women worldwide. Paternal consumption of a highfat diet has been shown to program breast cancer risk in female offspring. Orange juice is widely consumed and is known for its content of bioactive compounds that may have a role in regulating epigenetic processes. Therefore, the aim of the present study was to evaluate the effects of paternal obesity and orange juice consumption on female offspring susceptibility to chemically-induced breast carcinogenesis. Three-week-old C57BL/6 male mice were distributed in control (CO), control-orange juice (CJ), obese (OB) and obese-orange juice (OJ) groups, fed either a standard chow or a high-fat/high-sugar diet (45% lard-based diet supplemented with sweetened condensed milk), with water or orange juice, for 11 weeks before mating. Female offspring were weaned onto standard chow until 7 weeks of age and then were initiated with 7,12-dimethyl-benzo[a]anthracene to induce mammary tumors. CJ female offspring presented higher multiplicity of mammary tumors (p≤0.05) compared to CO offspring. Female offspring from OB group showed higher tumor latency (p≤0.05), lower tumor incidence (p≤0.05), higher multiplicity of tumors (p≤0.05), lower cell proliferation (Ki67) in the mammary ducts (p≤0.05) and lower global levels of H3K27me3 in the mammary gland (p≤0.05) when compared to CO offspring. No differences (p≥0,05) were observed between OB and OJ female offspring regarding these parameters. Consumption of orange juice by non-obese fathers during preconception increased susceptibility of female offspring to mammary carcinogenesis. Although paternal consumption of a high-fat/high-sugar diet during preconception decreased incidence and increased latency of tumors, the multiplicity of lesions increased. In addition, the data indicates that the response to orange juice consumption depends of the paternal metabolic context

O câncer de mama é o mais frequente entre as mulheres em todo o mundo. Foi demonstrado que o consumo paterno de uma dieta hiperlipídica aumenta o risco de câncer de mama nas filhas. O suco de laranja é amplamente consumido e é conhecido pelo seu conteúdo de compostos bioativos que podem ter um papel na regulação dos processos epigenéticos. Portanto, o objetivo do presente estudo foi avaliar os efeitos da obesidade paterna e do consumo de suco de laranja na suscetibilidade da prole feminina à carcinogênese mamária quimicamente induzida. Camundongos C57BL/6 machos com três semanas de idade foram distribuídos nos grupos controle (CO), controle de suco de laranja (CJ), obeso (OB) e obeso suco de laranja (OJ), alimentados com ração controle ou com ração hiperlipídica e hiperglicídica (45% das calorias proveniente de lipídeos, a base de banha de porco) suplementada com leite condensado, com água ou suco de laranja, durante 11 semanas antes do acasalamento. A prole feminina foi desmamada e recebeu ração controle até 7 semanas de idade e, então, foi iniciada com 7,12- dimetil-benzo[a]antraceno para induzir tumores mamários. A prole feminina CJ apresentou maior multiplicidade de tumores mamários (p≤0.05) em relação a prole feminina CO. A prole feminina OB apresentou maior latência tumoral (p≤0.05), menor incidência tumoral (p≤0.05), maior multiplicidade de tumores (p≤0.05), menor proliferação celular (Ki67) nos ductos mamários (p≤0.05) e menores níveis globais de H3K27me3 na glândula mamária (p≤0.05), quando comparada a prole feminina CO. Não foram observadas diferenças (p≥0,05) entre a prole feminina do OB e OJ em relação a esses parâmetros. O consumo de suco de laranja por pais não obesos durante o período pré-concepcional aumentou a susceptibilidade da prole feminina à carcinogênese mamária. Embora o consumo paterno de uma dieta hiperlipídica e hiperglicídica durante a preconcepção tenha diminuído a incidência e aumentado a latência, a multiplicidade dos tumores mamários aumentou. Ainda, os resultados indicam que a resposta ao consumo de suco de laranja depende do contexto metabólico paterno

Efeito da ingestão de sucos de laranja, variedades Moro e Pera, sobre o estresse oxidativo de camundongos saudáveis e com resistência à insulina induzida por dieta hiperlipídica e hiperglicídica / Effect of orange juices intake, cv. Moro and Pera, in oxidative stress of healthy and insulin resistant mice induced by high-fat and high carbohydrate diet

As laranjas, das variedades Moro e Pera, são conhecidas por sua composição de flavonoides, em especial, a subclasse flavanona, além das antocianinas na laranja Moro. Ambas as subclasses apresentam capacidade antioxidante, anti-inflamatória e hipolipidêmica, podendo atenuar as alterações metabólicas decorrentes do consumo de uma dieta hiperlipídica e hiperglicídica. Assim, o objetivo deste trabalho foi avaliar o efeito da ingestão de sucos de laranja, das variedades Moro e Pera, sobre os parâmetros oxidativos de camundongos saudáveis e com resistência à insulina, esta última induzida por uma dieta hiperlipídica e hiperglicídica. Camundongos machos, da linhagem C57Bl/6, foram distribuídos em seis grupos, três grupos receberam uma dieta controle e três grupos uma dieta hiperlipídica e hiperglicídica, durante 12 semanas. Concomitante às dietas, dois grupos de cada tratamento receberam suco de laranja Pera e o outro suco de laranja Moro em substituição à água de beber. Na 10ª e 11ª semana, foram realizados o ipITT e o ipGTT. No final do tratamento, foram coletados o sangue, fígado e coração. As flavanonas majoritárias caracterizadas em ambos os sucos foram a narirutina e hesperitina. Além das flavanonas, a cianidina 3-glucosídeo foi a antocianina majoritária no suco de laranja Moro. O consumo de dieta hiperlipídica e hiperglicídica acarretou em ganho de peso e adiposidade corporal, além de alterações metabólicas, como intolerância à insulina e a glicose, com o desenvolvimento da resistência insulínica, de acordo com o índice de HOMA-IR. Contudo, o suco de laranja, da variedade Pera, foi capaz de atenuar os parâmetros metabólicos como a sensibilidade à insulina, além da adiposidade e peso corporal. No tecido hepático, foi observado redução da peroxidação lipídica e da expressão proteica da enzima catalase nos animais tratados com dieta hiperlipídica e hiperglicídica, quando comparado à dieta controle; e aumento na atividade de superóxido dismutase no tecido cardíaco. Não se observou diferenças significativas nas demais enzimas antioxidantes, bem como no dano oxidativo ao DNA avaliado pelo ensaio do cometa. No tecido cardíaco, foi observado aumento da peroxidação lipídica e de glutationa peroxidase nos animais tratados com dieta controle mais suco de laranja Pera e suco de laranja Moro, respectivamente. Assim, a dieta hiperlipídica e hiperglicídica não acarretou em estresse oxidativo e, de maneira geral, os sucos não alteraram este quadro. Como conclusão, o suco de laranja Pera, atenuou as alterações metabólicas relacionadas ao metabolismo de carboidratos, mas não alterou os parâmetros de estresse oxidativo

Oranges, Moro and Pera varieties, are known for their composition of flavonoids, especially the subclass flavanone, beyond the anthocyanins in the Moro variety. Both subclasses present antioxidant, anti-inflammatory and hypolipidemic capacities, and may attenuate the metabolic alterations due to the consumption of a hyperlipidemic and hyperglycemic diet. Thus, the objective of this study was to evaluate the effect of orange juice intake, Moro and Pera varieties, on the oxidative parameters of insulin resistant mice induced by a high-fat and high carbohydrate diet. Mice (C57Bl/6 strain) were distributed in six groups: three groups received a control diet and three groups received a high-fat and high carbohydrate diet during 12 weeks. Concomitantly to the diets, two groups of each treatment received Pera and Moro orange juice in replacement of water. The ipITT and ipGTT were done at 11 e 12 week. At the ending of the treatment, blood, liver and heart were collected. The flavanones narirutine and hesperitin were the major flavanones in both juices. In addition, cyanidin 3-glycoside was the major anthocyanin in Moro orange juice. The consumption of high-fat and high carbohydrate diet resulted in weight gain and body adiposity, and metabolic alterations, such as decrease in insulin tolerance and development of insulin resistance, according to the HOMA-IR index. However, orange juice of the Pera variety was able to attenuate metabolic parameters increasing insulin sensitivity and decreasing adiposity and body weight. In hepatic tissue, decrease on lipid peroxidation and protein expression of the catalase were observed in animals treated with high-fat and high carbohydrate diet when compared to the control diet; and an increase activity of the superoxide dismutase in cardiac tissue. No significant differences were observed in the other antioxidant enzymes, as well as in the oxidative DNA damage assessed by the comet assay. In cardiac tissue, lipid peroxidation and glutathione peroxidase were increased in animals treated with control diet plus Pera and Moro orange juice, respectively. Thus, the high-fat and high carbohydrate diet did not promote oxidative stress and, in general, the juices did not alter oxidative parameters. In conclusion, Pera orange juice attenuate the metabolic alterations related to the metabolism of carbohydrates, but not alter oxidative stress parameters

Characterization of the Key Aroma Compounds in Five Varieties of Mandarins by Gas Chromatography-Olfactometry, Odor Activity Values, Aroma Recombination, and Omission Analysis.

In this study, volatile compounds of five varieties of mandarin juices [Tankan, Miyagawa, Mashui (MS), Skiranui, and Ponkan (PG)] were investigated by gas chromatography-olfactometry (GC-O) and gas chromatography-mass spectrometry (GC-MS). A total of 47 volatile compounds were identified by GC-MS. Partial least-squares regression was used to process the mean scores from sensory evaluation by panelists of volatile compounds and samples. The sample PG was associated with "fruity", "floral", and "sweet" notes, while MS was correlated with "green" and "peely" notes. In addition, 36 aroma-active compounds, including esters, alcohols, aldehydes, ketones, and monoterpenes, were detected by GC-O. According to the quantitative results, 29 aroma compounds were important, which indicated that their odor activity values (OAVs) were ≥1. On the basis of the GC-O results and OAVs of these volatile compounds, 22 odor-active compounds were mixed to simulate successfully the overall aroma of PG mandarin juice. Furthermore, omission experiments confirmed that nonanal, hexanal, linalool, and (R)-(+)-limonene were the key odorants for the overall aroma of PG juice sample and that ß-ionone, decanal, γ-terpinene, and methyl butyrate were also important odor-active compounds.

The high content of ß-carotene present in orange-pulp fruits of Carica papaya L. is not correlated with a high expression of the CpLCY-ß2 gene.

We investigated the transcriptional regulation of six genes involved in carotenoid biosynthesis, together with the carotenoid accumulation during postharvest ripening of three different papaya genotypes of contrasting pulp color. Red-pulp genotype (RPG) showed the lowest content of yellow pigments (YP), such as ß-cryptoxanthin, zeaxanthin, and violaxanthin, together with the lowest relative expression levels (REL) of CpLCY-ß2 and CpCHX-ß genes. On the contrary, the yellow-pulp genotype (YPG) showed the highest content of YP and the highest REL of CpLCY-ß2 and CpCHX-ß genes. Interestingly, the orange-pulp genotype (OPG) showed intermediate content of YP and intermediate REL of CpLCY-ß2 and CpCHX-ß genes. The highest content of ß-carotene shown by OPG despite having an intermediate REL of the CpLCY-ß2 genes, suggests a post-transcriptional regulation. Thus, the transcriptional level of the genes, directing the carotenoid biosynthesis pathway, can partially explain the accumulation of carotenoids during the postharvest ripening in C. papaya genotypes of contrasting pulp color.

Fresh-Squeezed Orange Juice Properties Before and During In Vitro Digestion as Influenced by Orange Variety and Processing Method.

This study aims to analyze the influence of processing and orange variety on initial quality, antioxidant activity, total polyphenol content (TPC), and ascorbic acid content of fresh-squeezed orange juice during in vitro digestion. Fresh-squeezed orange juice was made from Fukumoto Navel, Lane Late Navel, Olinda Nucellar Valencia, and Campbell Valencia oranges, and was processed thermally and nonthermally. Antioxidant activity (FRAP and ABTS assays), TPC (Folin-Ciocalteu method), and ascorbic acid were analyzed before and after gastrointestinal digestion. Bioaccessibility was calculated by comparing the measured values after digestion with the initial value for each juice. Orange variety significantly influenced pH, acidity, and total soluble solids (P < 0.0001). Antioxidant activity by FRAP was significantly higher (P < 0.0001) in Fukumoto Navel orange juice (16.0 ± 0.4 mM Trolox) than the other juices (range: 9.1 to 10 mM Trolox). TPC was significantly influenced by orange variety (P < 0.0001) and ranged from 521 ± 6 (Campbell Valencia) to 800 ± 11 mg gallic acid/L (Lane Late Navel). Processing method did not influence antioxidant and polyphenol bioaccessibility (P > 0.05). However, antioxidant activity by ABTS and ascorbic acid bioaccessibility were significantly influenced by orange variety (P < 0.0001). These results indicate that fruit variety and nutrient bioaccessibility should be considered to optimize processing and formulation parameters. PRACTICAL APPLICATION: Processing method and variety of fruit during beverage manufacturing may influence its nutrient bioaccessibility. We present a detailed examination of the influence of orange variety, harvest time, and processing method on initial quality parameters and antioxidant bioaccessibility. It is recommended that nutritional analysis of beverages take into consideration both the bioaccessibility of nutrients and the initial nutrient content of the beverage. These results can be used for juice processors to take into consideration the harvest season and produce variety to manufacture beverages with higher nutritional quality.

Differentiation Using Microwave Plasma Torch Desorption Mass Spectrometry of Navel Oranges Cultivated in Neighboring Habitats.

The molecular fingerprinting of intact fruit samples combined with statistical data analysis can allow the assessment of fruit quality and location of origin. Herein, microwave plasma torch desorption ionization mass spectrometry (MPT-MS) was applied to produce molecular fingerprints for the juice sac and exocarp of navel oranges cultivated in three closely located habitats, and the mass spectrometric fingerprints were differentiated by principal component analysis (PCA). Because of the relatively high temperature and high ionization efficiency of MPT, the volatile aroma compounds and semivolatile chemicals in the navel oranges were sensitively detected and confidently identified by collision induced dissociation (CID). The limit of detection (LOD) of MPT-MS for vanillin was 0.119 µg/L, with the relative standard deviation (RSD, n = 10) of 1.7%. The results showed that MPT-MS could be a powerful analytical platform for the sensitive molecular analysis of fruits at molecular level with high chemical specificity, allowing differentiation between the same sorts grown in neighboring habitats.

Laser-induced Fluorescence Spectroscopy (LIFS) for Discrimination of Genetically Close Sweet Orange Accessions ( Citrus sinensis L. Osbeck).

Although there is substantial diversity among cultivated sweet oranges genotypes with respect to morphological, physiological, and agronomic traits, very little variation at DNA level has been observed. It is possible that this low DNA molecular variability is due to a narrow genetic basis commonly observed in this citrus group. The most different morphological characters observed were originated through mutations, which are maintained by vegetative propagation. Despite all molecular tools available for discrimination between these different accessions, in general, low polymorphism has been observed in all groups of sweet oranges and they may not be identified by molecular markers. In this context, this paper describes the results obtained by using laser-induced fluorescent spectroscopy (LIFS) as a tool to discriminate sweet orange accessions ( Citrus sinensis L. Osbeck) including common, low acidity, pigmented, and navel orange groups, with very little variation at DNA level. The findings showed that LIFS combined with statistical methods is capable to discriminate different accessions. The basic idea is that citrus leaves have multiple fluorophores and concentration depends on their genetics and metabolism. Thus, we consider that the optical properties of citrus leaves may be different, depending on variety. The results have shown that the developed method, for the best classification rate, reaches an average sensitivity and specificity of 95% and 97.5%, respectively. An interesting application of this study is the development of an economically viable tool for early identification in seedling certification, in citrus breeding programs, in cultivar protection, or in germplasm core collection.

Identification, characterization and expression analysis of lineage-specific genes within sweet orange (Citrus sinensis).

BACKGROUND: With the availability of rapidly increasing number of genome and transcriptome sequences, lineage-specific genes (LSGs) can be identified and characterized. Like other conserved functional genes, LSGs play important roles in biological evolution and functions. RESULTS: Two set of citrus LSGs, 296 citrus-specific genes (CSGs) and 1039 orphan genes specific to sweet orange, were identified by comparative analysis between the sweet orange genome sequences and 41 genomes and 273 transcriptomes. With the two sets of genes, gene structure and gene expression pattern were investigated. On average, both the CSGs and orphan genes have fewer exons, shorter gene length and higher GC content when compared with those evolutionarily conserved genes (ECs). Expression profiling indicated that most of the LSGs expressed in various tissues of sweet orange and some of them exhibited distinct temporal and spatial expression patterns. Particularly, the orphan genes were preferentially expressed in callus, which is an important pluripotent tissue of citrus. Besides, part of the CSGs and orphan genes expressed responsive to abiotic stress, indicating their potential functions during interaction with environment. CONCLUSION: This study identified and characterized two sets of LSGs in citrus, dissected their sequence features and expression patterns, and provided valuable clues for future functional analysis of the LSGs in sweet orange.

Diversity in sensory quality and determining factors influencing mandarin flavor liking.

We evaluated the sensory quality of 42 mandarin varieties that belong to 7 different natural subgroups: common mandarin (Citrus reticulata Blanco), Clementine (Citrus clementina Hort. ex. Tan), Satsuma (Citrus unshiu Marcovitch), Mediterranean mandarin (Citrus deliciosa Tenore), King mandarin (Citrus nobilis Loureiro), and mandarin hybrids, such as tangor (Citrus reticulata × Citrus sinensis) and tangelo (Citrus reticulata × Citrus paradisi). Consumer flavor acceptance tests revealed wide diversity in flavor preferences among mandarin varieties and subgroups. Furthermore, descriptive flavor-analysis tests conducted with the aid of a trained sensory panel revealed that the 9 most preferred varieties had similar flavor profiles, characterized by high sweetness, moderate to low acidity levels, low bitterness and gumminess, strong fruity and mandarin flavor, and high juiciness. The average total soluble solids (TSS) and acidity levels among the highly preferred varieties were 13.1% and 1.1%, respectively. In contrast, the 8 least preferred varieties were either too sour or gummy or had low levels of sweetness, fruity, or mandarin flavor, and either high acidity levels (>1.4%) or low TSS levels (<12.0%). Pearson tests revealed significant positive correlations between flavor acceptance and perceptions of sweetness, fruitiness, and mandarin flavor, and negative correlations with acidity levels and perceptions of sourness, bitterness, and gumminess. Principle component analysis clearly distinguished between highly accepted varieties that were tightly correlated with high TSS levels and perceptions of sweetness fruitiness and mandarin flavor, as compared with the least accepted varieties that were correlated with high acidity levels and perceptions of sourness, bitterness, and gumminess.

Genome-wide identification and phylogenetic analysis of the AP2/ERF gene superfamily in sweet orange (Citrus sinensis).

Sweet orange (Citrus sinensis) plays an important role in the economy of more than 140 countries, but it is grown in areas with intermittent stressful soil and climatic conditions. The stress tolerance could be addressed by manipulating the ethylene response factor (ERF) transcription factors because they orchestrate plant responses to environmental stress. We performed an in silico study on the ERFs in the expressed sequence tag database of C. sinensis to identify potential genes that regulate plant responses to stress. We identified 108 putative genes encoding protein sequences of the AP2/ERF superfamily distributed within 10 groups of amino acid sequences. Ninety-one genes were assembled from the ERF family containing only one AP2/ERF domain, 13 genes were assembled from the AP2 family containing two AP2/ERF domains, and four other genes were assembled from the RAV family containing one AP2/ERF domain and a B3 domain. Some conserved domains of the ERF family genes were disrupted into a few segments by introns. This irregular distribution of genes in the AP2/ERF superfamily in different plant species could be a result of genomic losses or duplication events in a common ancestor. The in silico gene expression revealed that 67% of AP2/ERF genes are expressed in tissues with usual plant development, and 14% were expressed in stressed tissues. Because the AP2/ERF superfamily is expressed in an orchestrated way, it is possible that the manipulation of only one gene may result in changes in the whole plant function, which could result in more tolerant crops.

Comparison of carotenoid accumulation and biosynthetic gene expression between Valencia and Rohde Red Valencia sweet oranges.

Carotenoid accumulation and biosynthetic gene expression levels during fruit maturation were compared between ordinary Valencia (VAL) and its more deeply colored mutant Rohde Red Valencia orange (RRV). The two cultivars exhibited different carotenoid profiles and regulatory mechanisms in flavedo and juice sacs, respectively. In flavedo, there was uncoordinated carotenoid accumulation and gene expression in RRV during green stages, which might be related to the expression of certain gene(s) in the MEP (methylerythritol phosphate) pathway. The carotenoid biosynthesis pathway shifting from α,ß-xanthophylls to ß,ß-xanthophylls synthesis occurred in RRV earlier than VAL during orange stages. In juice sacs, the low carotenoid content in both cultivars coincided with low expression of LCYE-Contig03 and LCYE-Contig24 during green stages, suggesting LCYE might be a limiting step for carotenoid accumulation. VAL mainly accumulated violaxanthin, but RRV accumulated ß-cryptoxanthin and violaxanthin during orange stages, which corresponded to differences in juice color. Several upstream genes (PDS-Contig17, LCYB-Contig19, and ZDS members) and a downstream gene (ZEP) were expressed at higher levels in RRV than VAL, which might be responsible for greater accumulation of ß-cryptoxanthin and violaxanthin in RRV, respectively.

Genome-wide analysis of the R2R3-MYB transcription factor gene family in sweet orange (Citrus sinensis).

MYB transcription factor represents one of the largest gene families in plant genomes. Sweet orange (Citrus sinensis) is one of the most important fruit crops worldwide, and recently the genome has been sequenced. This provides an opportunity to investigate the organization and evolutionary characteristics of sweet orange MYB genes from whole genome view. In the present study, we identified 100 R2R3-MYB genes in the sweet orange genome. A comprehensive analysis of this gene family was performed, including the phylogeny, gene structure, chromosomal localization and expression pattern analyses. The 100 genes were divided into 29 subfamilies based on the sequence similarity and phylogeny, and the classification was also well supported by the highly conserved exon/intron structures and motif composition. The phylogenomic comparison of MYB gene family among sweet orange and related plant species, Arabidopsis, cacao and papaya suggested the existence of functional divergence during evolution. Expression profiling indicated that sweet orange R2R3-MYB genes exhibited distinct temporal and spatial expression patterns. Our analysis suggested that the sweet orange MYB genes may play important roles in different plant biological processes, some of which may be potentially involved in citrus fruit quality. These results will be useful for future functional analysis of the MYB gene family in sweet orange.