Evaluation of citrus somatic hybrids for tolerance to Phytophthora nicotianae and citrus tristeza virus

Somatic hybridization is a biotechnology tool that can be used in citrus breeding programs to produce somatic hybrids with the complete genetic combination of both parents. The goal of this work was to test the reaction of citrus somatic hybrids that may be useful as rootstocks to trunk and root infections caused by Phytophthora nicotianae van Breda de Haan (P parasitica Dastur) and to citrus tristeza virus (CTV). The somatic hybrids evaluated were `Caipira` sweet orange (Citrus sinensis L. Osbeck) + `Rangpur` lime (C. limonia Osbeck), `Caipira` sweet orange + `Cleopatra` mandarin (C. reshni hort. ex Tanaka), `Caipira` sweet orange + `Volkamer` lemon (C. volkameriana V Ten. & Pasq.), `Caipira` sweet orange + rough lemon (C. jambhiri Lush.), `Cleopatra` mandarin + `Volkamer` lemon, `Cleopatra` mandarin + sour orange (C. aurantium L.), `Rangpur` lime + `Sunki` mandarin (C. sunki (Hayata) hort. ex Tanaka), `Ruby Blood` sweet orange (C. sinensis L. Osbeck) + `Volkamer` lemon, `Rohde Red` sweet orange (C. sinensis L. Osbeck) + `Volkamer` lemon, and `Valencia` sweet orange + Fortunella obovata hort. ex Tanaka. For P. nicotianae trunk and root infection assays, plants of the somatic hybrids, obtained from 9-month semi-hardwood cuttings, were evaluated and compared with diploid citrus rootstock cultivars after mycelia inoculation in the trunk or spore infestation in the substrate, respectively. `Cleopatra` mandarin + sour orange, `Rangpur` lime + `Sunki` mandarin, `Cleopatra` mandarin + `Volkamer` lemon, `Ruby Blood` sweet orange + `Volkamer` lemon, `Rohde Red` sweet orange + `Volkamer` lemon, and `Caipira` sweet orange + `Volkamer` lemon had less trunk rot occurrence, whereas the somatic hybrids `Cleopatra` mandarin + `Volkamer` lemon, `Cleopatra` mandarin + sour orange, `Caipira` sweet orange + `Volkamer` lemon, and `Caipira` sweet orange + `Rangpur` lime were tolerant to root rot. For CTV assays, plants of the somatic hybrids along with tolerant and intolerant rootstocks were budded with a mild strain CTV-infected or healthy `Valencia` sweet orange budwood. Differences in average scion shoot length indicated that the hybrids `Cleopatra` mandarin + sour orange and `Valencia` sweet orange + Fortunella obovata were intolerant to CTV (c) 2007 Elsevier B.V. All rights reserved.

Effect of Citrus floral extracts on the foraging behavior of the stingless bee Scaptotrigona pectoralis (Dalla Torre)

Effect of Citrus floral extracts on the foraging behavior of the stingless bee Scaptotrigona pectoralis (Dalla Torre). Stingless bees have an important role as pollinators of many wild and cultivated plant species in tropical regions. Little is known, however, about the interaction between floral fragrances and the foraging behavior of meliponine species. Thus we investigated the chemical composition of the extracts of citric (lemon and orange) flowers and their effects on the foraging behavior of the stingless bee Scaptotrigona pectoralis. We found that each type of flower has its own specific blend of major compounds: limonene (62.9%) for lemon flowers, and farnesol (26.5%), (E)-nerolidol (20.8%), and linalool (12.7%) for orange flowers. In the foraging experiments the S. pectoralis workers were able to use the flower extracts to orient to the food source, overlooking plates baited with hexane only. However, orange flower extracts were seemingly more attractive to these worker bees, maybe because of the particular blend present in it. Our results reveal that these fragrances are very attractive to S. pectoralis, so we can infer that within citric orchards they could be important visitors in the study area; however habitat destruction, overuse of pesticides and the competitive override by managed honeybees might have put at risk their populations and thus the ecological services they provide to us.

Polyembryony and identification of Volkamerian lemon zygotic and nucellar seedlings using RAPD

The objectives of this work were to evaluate the frequency of polyembryony, and to identify zygotic and nucellar seedlings of Citrus volkameriana using RAPD. Twenty-five polyembryonic and eight monoembryonic seeds were cultivated in vitrofor six months. DNA from seedlings was extracted and used in combination with five RAPD primers to identify zygotic or nucellar origin of the seedlings. Environmental conditions of the year affected significantly (P<0.05) the morphological characteristics of fruitsand the number ofembryos per seed. Polyembryonic seeds ranged from 30.9%, 44.8% to 54.4% over three years. Morphological characteristic was not correlated with polyembryony. In vitro culture enable all embryos of each seed to grow, favoring the percentage of seedlings identified as zygotic. In polyembryonic and monoembryonic seeds, 25.9% and 87.5% of the seedlings, respectively, were sexually originated. In polyembryonic seeds, not all zygotic seedlings were produced by small embryos located at the micropyle.

Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts

The objective of this study was to produce citrus somatic asymmetric hybrids by fusing gamma-irradiated protoplasts with iodoacetamide-treated protoplasts. Protoplasts were isolated from embryogenic suspension cells of grapefruit (Citrus paradisi Macfad.) cultivars Ruby Red and Flame, sweet oranges (C. sinensis Osbeck) 'Itaboraí', 'Natal', Valencia', and 'Succari', from 'Satsuma' (C. unshiu Marcow.) and 'Changsha' mandarin (C. reticulata Blanco) and 'Murcott' tangor (C. reticulata x C. sinensis). Donor protoplasts were exposed to gamma rays and receptor protoplasts were treated with 3 mmol L-1 iodoacetamide (IOA), and then they were fused for asymmetric hybridization. Asymmetric embryos were germinated, and the resulting shoots were either grafted onto sour orange, rough lemon or 'Swingle' (C. paradisi x Poncirus trifoliata) x 'Sunki' mandarin rootstock seedlings, or rooted after dipping their bases in indol-butyric acid (IBA) solution. The products were later acclimatized to greenhouse conditions. Ploidy was analyzed by flow cytometry, and hybridity was confirmed by amplified fragment length polymorphism (AFLP) analysis of plantlet DNAsamples. The best treatment was the donor-recipient fusion combination of 80 Gy-irradiated 'Ruby Red' protoplasts with 20 min IOA-treated 'Succari' protoplasts. Tetraploid and aneuploid plants were produced. Rooting recalcitrance was solved by dipping shoots' stems in 3,000 mg L-1 IBA solution for 10 min.

Monitoring the viability of citrus rootstocks seeds stored under refrigeration

The citrus nursery tree is produced through the bud grafting process, in which rootstock is usually grown from seed germination. The objective of this research was to evaluate, in two dissimilar environmental conditions, the viability and polyembryony expression of five citrus rootstocks seeds stored in different periods under refrigeration. The rootstock varieties evaluated were: Rangpur lime (Citrus limonia Osb. cv. Limeira), Trifoliate orange (Poncirus trifoliata Raf. cv. Limeira), Citrumelo (P. trifoliata x C. paradisi Macf. cv. Swingle), Sunki mandarin (C. sunki Hort. ex Tanaka) and Volkamer lemon (C. volkameriana Ten. & Pasq. cv. Catania 2). The experimental design was the randomized blocks in a 11 x 5 x 2 factorial scheme, evaluating from time zero to the tenth month of storage, the five varieties of rootstock in two environments: germination and growth B.O.D type chamber (Biological Oxygen Demand - Eletrolab Brand Model FC 122) at 25 °C; and greenhouse seedbed with partial temperature control (22 °C to 36 °C) and humidity control (75-85%). The plot had 24 seeds in four replicates, using trays with substrate in greenhouse and Petri dishes with filter paper in B.O.D. chamber. The seed germination rate and polyembryony expression were evaluated monthly. It was concluded that Trifoliate and Citrumelo Swingle seeds can be stored for up to seven months, while Volkamer lemon, Rangpur lime and Sunki seeds can be stored for up to ten months. The polyembryony expression rate was slightly higher when measured in greenhouse than in B.O.D. chamber and remained stable in both environments until the seventh month, from which dropped sharply. Citrumelo Swingle seeds expressed the highest polyembryony rate (18.8%), followed by Rangpur lime and Volkamer lemon (average value of 13.7%), Sunki (9.4%) and Trifoliate (3.2%). Despite some differences among varieties, the viability of rootstock stored seeds can be monitored either in the greenhouse or in B.O.D. germination chamber, the latter being the faster and more suitable method.

Atividade antioxidante do extrato de sementes de limão (Citrus limon) adicionado ao óleo de soja em teste de estocagem acelerada

The aim of this work was to evaluate antioxidant activity of lemon seeds added to soybean oil, submitted to accelerated incubator-storage test and to determine its synergistic effect with the synthetic antioxidant TBHQ. The treatments Control, TBHQ (50 mg/kg), LSE (2,400 mg/kg Lemon Seed Extract), Mixture 1 (LSE + 50 mg/kg TBHQ) and Mixture 2 (LSE + 25 mg/kg TBHQ) were prepared and subjected to the accelerated incubator-storage test at 60 ºC for 12 days; samples were taken every 3 days and analyzed regarding peroxide value and conjugated dienes. The results showed that antioxidant activity of the tested treatments were: TBHQ = Mixture 1 = Mixture 2 > LSE > Control.

Woody Gall symptoms in different citrus species and varieties artificially colonized by the citrus brown aphid Toxoptera citricidus

Seedlings of 41 different citrus species and varieties were massively colonized with the citrus brown aphid Toxoptera citricidus, obtained from Pêra sweet orange (Citrus sinensis) trees, presenting symptoms of the "Capão Bonito" complex of the Citrus tristeza virus (CTV). The objective was to evaluate resistance or tolerance of the varieties to that virus complex, but even after eight months of inoculation no stem pitting was observed in the plants. Otherwise, the presence of galls similar to those induced by the vein enation-woody gall disease was observed in 73% of the plants of Volkamer Palermo (Citrus volkameriana), 60% of the Volkamer Catania 2, 2% of the Rangpur Lime D.22.30 (Citrus limonia), 13% of the Volkamer Australian Red, the Rangpur Lime hybrid, the Orlando tangelo (Citrus reticulata x Citrus paradisi) and the Florida Rough lemon (C. jambhiri), and 7% of the Carrizo citrange (Poncirus trifoliata x Citrus sinensis). The highest incidence and the largest gall size were observed in the Palermo Volkamer showing that this clone would be the most suitable to be used as an indicator plant in biological indexing tests for the disease. There are no previous reports in the literature about the occurrence of woody galls in Orlando tangelo and Carrizo citrange.

Osmotic priming methodologies in relation to the physiological performance of rangpur lime seeds (Citrus limonia Osbeck)

Several mechanisms have been used to promote rapid germination of citrus seeds and uniform seedling emergence. We evaluated the effects of osmotic priming on the physiological performance of Rangpur lime seeds (Citrus limonia Osbeck). Seeds were treated with 30 g of Captan and 10 g of Tecto 600 in 20-litre batches and stored, without drying, at 10 ºC and 50% relative humidity for periods of 3, 6 and 9 months. After each period, seeds were primed at 25 ºC, in the light, by immersion in Poliethylenoglicol (PEG 6000), potassium nitrate (KNO3) and 70% PEG 6000 plus 30% KNO3, all at an osmotic potential of -1.1MPa, for priming periods of 3, 6, 9 and 12 days. Percentage germination, tray emergence and the emergence rate index (ERI) were evaluated. Priming in PEG 6000 solution, independent of priming period, or in KNO3 or PEG 6000 plus KNO3 for up to 9 days, were efficient at improving the physiological performance of seeds stored for up to 3 months. Osmotic priming appears to be a promising technique for improving the physiological quality of Rangpur lemon seeds.

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)

Peroxidase activity and total phenol content in citrus cuttings treated with different copper sources

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Phytochemicals and Antioxidant Activity of Citrus Seed Oils

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

Potencial antioxidante de extratos de sementes de limão (Citrus limon)

The present study aimed at evaluating the behavior of the soybean oil with the addition of lemon seed extracts (Citrus limon), in different concentrations, through oxidative stability and also at measuring the antioxidant activity using the DPPH free radical method and total phenolic compounds. The concentration of 2,400 mg.kg(-1) for the lemon seed extract, galego variety, was the one that provided the soybean oil with the best oxidative stability. The maximum antioxidant activities and the concentration values of the natural extract sufficient to obtain 50% of maximum effect, estimated at 100% (EC(50)), determined by DPPH for the extract and gallic acid were 70.58%, 69.94 mu g.mL(-1) and 75.07%, 64.73 mu g.mL(-1), respectively. The concentration of total phenolic compounds, determined by the Folin-Ciocalteu method, was of 76 mg.g(-1). Thus, it can be said that the lemon seed extract, galego, presents natural antioxidant action demonstrating potential to be used in foods.

Atividade antioxidante do extrato de sementes de limão (Citrus limon) adicionado ao óleo de soja em teste de estocagem acelerada

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biologia de Polyphagotarsonemus latus (Banks) (Acari: Tarsonemidae) em limão siciliano (Citrus limon Burm)

No estudo da biologia de Polyphagotarsonemus latus em limão Siciliano, foram utilizados potes plásticos circulares com capacidade de 250 ml, contendo areia esterilizada como suporte para dois frutos novos com aproximadamente 2,0 cm de diâmetro. O ensaio foi conduzido a 27,1 ± 0,5°C, umidade relativa de 67,6 ± 1,3% e fotofase contínua. O período de ovo a adulto durou 3,7 ± 0,1 dias para fêmeas e 3,6 ± 0,1 dias para machos, com sobrevivência de 100%. Após um período de pré-oviposição de 1,0 ± 0,2 dias, as fêmeas depositaram 5,6 ± 0,5 ovos por dia durante 10,5 ± 0,9 dias, totalizando 58,9 ± 6,7 ovos por fêmea. A longevidade foi de 13,4 ± 1,0 dias para fêmeas e 12,0 ± 2,4 dias para machos. A razão intrínseca de aumento (rm) foi de 0,359, a razão finita de aumento (l) de 1,43 indivíduos por fêmea por dia, o tempo médio de uma geração (T) de 10,34 dias e a taxa líquida de reprodução (Ro) de 41,0.