Metabolite profiles of essential oils in citrus peels and their taxonomic implications

China is an important center of origin for the genus Citrus L. of the family Rutaceae and is rich in wild Citrus species. The taxonomy of Citrus has been a subject of controversy for more than a half century. We propose that the metabolite profiles of Chinese native Citrus species can be used for classification and understanding of the taxonomic relationships within the Citrus germplasm. In this study, triplicate gas chromatography-mass spectrometry (GC-MS) metabolite profiles of 20 Citrus species/varieties were acquired, including 10 native varieties originating in China. R-(+)-limonene, alpha-pinene, sabinene and alpha-terpinene were found to be major characteristic components of the essential oils analyzed in this study, and these compounds contributed greatly to the metabolic classification. The three basic species of the subgenus Eucitrus (Swingle's system), i.e., C. reticulata Blanco, C. medica L. and C. grandis Osb., were clearly differentiated based upon their metabolite profiles using hierarchical cluster analysis (HCA) and partial least square-discriminant analysis (PLS-DA). All the presumed hybrid genotypes, including sweet orange (C. sinensis Osb.), sour orange (C. aurantium L.), lemon (C. limon Burm.f.), rough lemon (C. jambhiri Lush.), rangpur lime (C. limonia Osb.) and grapefruit (C. paradisi Macf.), were grouped closely together with one of their suggested parent species in the HCA-dendrogram and the PLS-DA score plot. These results clearly demonstrated that the metabolite profiles of Citrus species could be utilized for the taxonomic classification of the genus and are complementary to the existing taxonomic evidence, especially for the identification and differentiation of hybrid species.

Low glyphosate rates do not affect Citrus limonia (L.) Osbeck seedlings

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito de herbicidas residuais, aplicados por vários anos consecutivos, na distribuição do sistema radicular da laranjeira Natal (Citrus sinensis.(L.) osbeck)

O presente trabalho foi desenvolvido com o objetivo de verificar a influência do uso contínuo de vários herbicidas residuais, sobre a distribuição das radicelas de laranjeiras Natal (Citrus sinensis (L.) Osbeck) enxertadas sobre limão Cravo (Cithus limonia Osbeck) e plantadas em janeiro de 1970. O experimento foi instalado no pomar de produção da Faculdade de Ciências Agrárias e Veterinárias do Campus de Jaboticabal (UNESP), em Latossol Roxo distrófico, com 1,6% de matéria orgânica e 51% de argila. O delineamento experimental utilizado foi o de blocos ao acaso, dentro de um esquema fatorial 11x 3x2x2, com três repetições. Os tratamentos utilizados, com as respectivas doses em kg do i.a/ha foram: fluometuron a 4,2; simazíne a 4,8; atrazine a 4,8; bromacil a 3,2;bromacil (40%) + diuron (40%) a 4,8; bromacil (53,3%) + diuron (27,6%) a 4,8; terbacil a 3,2; oxadiazon a 1,5; dichlobenil a 3,0 e 6,0; além de uma testemunha capinada. O estudo das radicel as foi feito pelo método do trado, tendo sido tomadas amostras de solo contendo radicelas, nas distâncias de 80, 160 e 240 cm do tronco, nas camadas de 0 a 15 cm e 15 a 30 cm de profundidade e em duas direções distintas (entrelinhas e entreplantas). Os resultados mostraram que as radicelas localizam-se superficialmente, com 70% delas nos primeiros 15 cm, do total encontrado na camada superficial de 0 a 30 cm de profundidade do solo. Cerca de 75% das radicelas situam-se até a distância de 160 cm do tronco. Na camada mais superficial do solo (0 a 15 cm), a quantidade de radicelas diminuiu de maneira acentuada, à medida que se afastou do tronco. Na camada de 15 a 30 cm de profundidade, ocorreu uma distribuição horizontal mais uniforme, comparada com a da camada de 0 a 15 cm. As radicelas das plantas do cultivar Natal distribuiram-se uniformemente, tanto na direção das plantas da linha vizinha, quanto nas das plantas de uma mesma linha, para o espaçamento de 7,0 m x 7,2 m. Não se detectaram diferenças estatisticamente significativas nas quantidades de radicelas entre as parcelas dos vários tratamentos com herbicidas, excluindo-se desta forma, qualquer efeito negativo de tais produtos químicos sobre o sistema radicular das laranjeiras.

Efeito de diferentes porta-enxertos no vigor e produtividade da laranjeira 'pera' (Citrus sinensis L.) Osbeck)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Tolerância à dessecação e influência do tegumento na germinação de sementes de citrumelo 'swingle' (Citrus paradisi MACF X Poncirus trifoliata (L) RAF.)

O trabalho teve como objetivo estudar a tolerância à dessecação e a influência do tegumento na germinação de sementes de citrumelo 'Swingle'. As sementes foram extraídas manualmente e, em seguida, foi determinado o grau de umidade das sementes. Foi retirada uma amostra referente ao tratamento com o maior grau de umidade (48%) a ser estudado, e as demais foram submetidas à secagem em estufa com circulação forçada de ar (32±2ºC), visando à obtenção dos outros tratamentos com diferentes graus de umidade. O teste de germinação foi instalado em delineamento experimental inteiramente casualizado, num fatorial 6x2 (grau de umidade x presença ou ausência de tegumento), com quatro repetições de 25 sementes por parcela. Após a obtenção de cada tratamento as sementes foram tratadas com o fungicida Thiabendazole (0,4g.kg-1), semeadas em folhas de papel toalha umedecidas e confeccionados rolos que foram mantidos em câmara de germinação a 25ºC sob luz constante. As avaliações foram realizadas a cada sete dias até o 35º dia, sendo determinadas as porcentagens de germinação na primeira contagem, plântulas anormais, sementes mortas, sementes dormentes e germinação total. Também foram calculados o tempo médio e o índice de velocidade de germinação (IVG). Os dados foram submetidos à análise de variância, e as médias, comparadas pelo teste Tukey, a 5% de probabilidade. As sementes toleraram a dessecação até baixos níveis de umidade (16%), e a retirada do tegumento favoreceu o processo germinativo em sementes de citrumelo 'Swingle'.

Anxiolytic and sedative effects of extracts and essential oil from Citrus aurantium L.

Citrus aurantium L. is commonly used as an alternative treatment for insomnia, anxiety and epilepsy. Essential oil from peel (EOP) and hydroethanolic (70% w/v) extract (HE) from leaves were obtained. Hexanic (HF), dichloromethanic (DF) and final aqueous (AF) fractions were obtained from HE by successive partitions. Swiss male mice (35-45 g) were treated orally with 0.5 or 1.0 g/kg of these preparations 30 min before the experiments for the evaluation of the sedative/hypnotic activity (sleeping time induced by sodium pentobarbital-SPB: 40 mg/kg, i.p.), anxiolytic activity (elevated plus maze-EPM) and anticonvulsant activity (induced by pentylenetetrazole-PTZ: 85 mg/kg, se or by maximal electroshock-MES: 50 mA, 0.11s, corneal). The results showed that EOP (0.5 g/kg) increased the latency period of tonic seizures in both convulsing experimental models. This effect was not dose-dependent. Treatment with 1.0 g/kg increased the sleeping time induced by barbiturates and the time spent in the open arms of the EPM. Specific tests indicated that the preparation, in both doses used, did not promote deficits in general activity or motor coordination. HF and DF fractions (1.0 g/kg) did not interfere in the epileptic seizures, but were able to enhance the sleeping time induced by barbiturates. The results obtained with EOP in the anxiety model, and with EOP, HF and DF in the sedation model, are in accord with the ethnopharmacological use of Citrus aurantium L., which could be useful in primary medical care, after toxicological investigation.

Effects of the essential oil from Citrus aurantium L. in experimental anxiety models in mice

Citrus aurantium L. is popularly used to treat anxiety, among other indications suggesting central nervous system action. Previous studies showed anxiolytic effect in the essential oil from peel in mice evaluated on the elevated plus maze [Carvalho-Freitas, M.I.R., Costa, M., 2002. Anxiolytic and sedative effects of extracts and essential oil from Citrus aurantium L. Biological and Pharmaceutical Bulletin 25, 1629-1633.]. In order to better characterize the activity of the essential oil, it was evaluated in two other experimental models: the light-dark box and the marble-burying test, respectively related to generalized anxiety disorder and to obsessive compulsive disorder. Mice were treated acutely by oral route 30 min (single dose) or once a day for 15 days (repeated doses) before experimental procedures. In light-dark box test, single treatment with essential oil augmented the time spent by mice in the light chamber and the number of transitions between the two compartments. There were no observed alterations in the parameters evaluated in light-dark box after repeated treatment. Otherwise, single and repeated treatments with essential oil were able to suppress marble-burying behavior. At effective doses in the behavioral tests, mice showed no impairment on rotarod procedure after both single and repeated treatments with essential oil, denoting absence of motor deficit. Results observed in marble-burying test, related to obsessive compulsive disorder, appear more consistent than those observed in light-dark box. (c) 2005 Elsevier B.V. All rights reserved.

Effect of Citrus aurantium l. essential oil on muscle regeneration in mdx mice

Duchenne muscular dystrophy (DMD) is a severe X-linked recessive disorder characterized by the progressive loss of muscular strength. Mdx mutant mice show a marked deficiency in dystrophin, which was related to muscle membrane stability. The aim of this study was to verify the possible protective anti-inflammatory effect of citrus oil on mdx muscle fibers. Thus, adult male and female mdx mice (014/06-CEEA) were divided into control and citrus-treated. After 60 days of treatment, one ml of blood was collected for creatine kinase (CK) test. Diaphragm, sternomastoideus, anterior tibial and gastrocnemius muscles were removed and processed according to histological routine methods. The observed alterations indicate a direct effect of citrus. Recent studies have improved the diagnosis of muscular diseases but with no definitions of efficient treatments. Intervention with several therapies is important to many patients presenting muscular dystrophy, which enables them to live longer and be more active, while there is no development of gene therapies.

Gastric Ulcers in Middle-Aged Rats: The Healing Effect of Essential Oil from Citrus aurantium L. (Rutaceae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Polinização em cultura de laranja (Citrus sinensis L. Osbeck, var. Pera-Rio)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isoenzymatic polymorphism in Citrus spp. and Poncirus trifoliata (L.) Raf. (Rutaceae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development and characterization of polymorphic microsatellite markers for the sweet orange (Citrus sinensis L. Osbeck)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mites (Arachnida, Acari) on Citrus sinensis L. Osbeck orange trees in the state of Amazonas, Northern Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evaluation of microsatellite markers in cultivars of sweet orange (Citrus sinensis [L.] Osbeck)

Sweet orange is considered a very important species in the citrus world market and presents wide morphological variability. However, its characterization at the molecular level by random amplified polymorphic DNA (RAPD) and isozyme markers is not appropriate. Microsatellite or simple sequence repeats (SSRs) have been suggested as ideal for studies in cultures of vegetative propagation and as value markers for mapping in several species. However, information on microsatellite polymorphism in citrus species is scarce. In this work, microsatellite markers (AG-repeats) were developed from an enrichment library of genomic DNA of sweet orange cv. Pera (Citrus sinensis [L.] Osbeck), and 31 cultivars of sweet orange were evaluated. Evaluation of 18 microsatellite primers did not permit differentiation of the varieties studied. New microsatellite primers are being evaluated with the aim of detecting polymorphisms among the cultivars and closely related species to be used in genetic mapping programs.

Citrus aurantium L. essential oil exhibits anxiolytic-like activity mediated by 5-HT1A-receptors and reduces cholesterol after repeated oral treatment

Background: The current treatments for anxiety disorders and depression have multiple adverse effects in addition to a delayed onset of action, which has prompted efforts to find new substances with potential activity in these disorders. Citrus aurantium was chosen based on ethnopharmacological data because traditional medicine refers to the Citrus genus as useful in diminishing the symptoms of anxiety or insomnia, and C. aurantium has more recently been proposed as an adjuvant for antidepressants. In the present work, we investigated the biological activity underlying the anxiolytic and antidepressant effects of C. aurantium essential oil (EO), the putative mechanism of the anxiolytic-like effect, and the neurochemical changes in specific brain structures of mice after acute treatment. We also monitored the mice for possible signs of toxicity after a 14-day treatment.Methods: The anxiolytic-like activity of the EO was investigated in a light/dark box, and the antidepressant activity was investigated in a forced swim test. Flumazenil, a competitive antagonist of benzodiazepine binding, and the selective 5-HT1A receptor antagonist WAY100635 were used in the experimental procedures to determine the mechanism of action of the EO. To exclude false positive results due to motor impairment, the mice were submitted to the rotarod test.Results: The data suggest that the anxiolytic-like activity observed in the light/dark box procedure after acute (5 mg/kg) or 14-day repeated (1 mg/kg/day) dosing was mediated by the serotonergic system (5-HT1A receptors). Acute treatment with the EO showed no activity in the forced swim test, which is sensitive to antidepressants. A neurochemical evaluation showed no alterations in neurotransmitter levels in the cortex, the striatum, the pons, and the hypothalamus. Furthermore, no locomotor impairment or signs of toxicity or biochemical changes, except a reduction in cholesterol levels, were observed after treatment with the EO.Conclusion: This work contributes to a better understanding of the biological activity of C. aurantium EO by characterizing the mechanism of action underlying its anxiolytic-like activity. © 2013 Costa et al; licensee BioMed Central Ltd.