Somatic hybridization between Citrus sinensis (L.) Osbeck and C. grandis (L.) Osbeck

This work had as objective to produce citrus somatic hybrids between sweet oranges and pummelos. After chemical fusion of sweet orange embryogenic protoplasts with pummelo mesophyll-derived protoplasts, plants were regenerated by somatic embryogenesis and acclimatized in a greenhouse. The hybrids of 'Hamlin' sweet orange + 'Indian Red' pummelo and 'Hamlin' sweet orange + 'Singapura' pummelo were confirmed by leaf morphology, chromosome counting and molecular analysis. These hybrids have potential to be used directly as rootstocks aiming blight, citrus tristeza virus, and Phytophthora-induced disease tolerance, as well as for rootstocks improvement programs.

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.

Faunistic analysis of Carabidae and Staphylinidae (Coleoptera) in five agroecosystems in northeastern São Paulo state, Brazil

The objective of this study was to determined species composition and community structure of Carabidae and Staphylinidae in five areas of forest fragment and soybean/corn crops or orange orchard, from December 2004 to May 2007. Beetles were captured in pitfall traps distributed along two parallel transects of 200 m in length, placed across crop land/forest boundary fragment, with 100 m each. The Shannon-Wiener diversity and evenness indexes and Morisita similarity index were calculated. The carabids Abaris basistriatus Chaudoir, Calosoma granulatum Perty, Megacephala brasiliensis Kirby, Odontochila nodicornis (Dejean) and Selenophorus seriatoporus Putzeys. are dominant and are widely distributed in northeastern São Paulo state, Brazil. Point-scale species diversity was greatest at the transition between forest fragment and cultivated area. The carabid and staphylinid communities of the forest fragment were more similar to the community of orange orchard than that of soybean/corn crops.

Stability of Citrus tristeza virus protective isolates in field conditions

The objective of this work was to monitor the maintenance of Citrus tristeza virus (CTV) protective isolates stability in selected clones of 'Pêra' sweet orange (Citrus sinensis), preimmunized or naturally infected by the virus, after successive clonal propagations. The work was carried out in field conditions in the north of Paraná State, Brazil. Coat protein gene (CPG) analysis of 33 isolates collected from 16 clones of 'Pêra' sweet orange was performed using single strand conformational polymorphism (SSCP). Initially, the isolates were characterized by symptoms of stem pitting observed in clones. Then viral genome was extracted and used as template for the amplification of CPG by reverse transcription polimerase chain reaction (RTPCR). RTPCR products electrophoretic profiles were analyzed using the Jaccard coefficient and the UPGMA method. The majority of the clones had weak to moderate stem pitting symptoms and its CTV isolates showed alterations in the SSCP profiles. However, the stability of the protective complex has been maintained, except for isolates from two analised clones. Low genetic variability was observed within the isolates during the studied years.

Performance of 'Oneco' mandarin on six rootstocks in South Brazil

The objective of this work was to assess the effect of six rootstocks on yield, fruit quality, and growth of 'Oneco' mandarin during the first seven harvesting seasons, in Butiá, Rio Grande do Sul State, Brazil. The rootstocks evaluated were: 'Swingle' citrumelo (Citrus paradisi × Poncirus trifoliata), 'Caipira' orange (C. sinensis), 'Troyer' citrange (C. sinensis × P. trifoliata), 'Rangpur' lime (C. limonia), 'Volkamer' lemon (C. volkameriana), and 'Flying Dragon' trifoliata orange (P. trifoliata var. monstrosa). Plants budded onto 'Flying Dragon' had the lowest vegetative development, which indicates the dwarfing characteristics of this rootstock, and had the highest mean production efficiency, despite low yield. Plants grafted on 'Volkamer' lemon and 'Rangpur' lime had the highest alternate bearing. Under the experimental conditions evaluated, the most adequate rootstocks for mandarin 'Oneco' are 'Swingle' citrumelo and 'Troyer' citrange, regarding fruit yield and quality.

Genetic transformation of sweet oranges with the D4E1 gene driven by the AtPP2 promoter

The objective of this work was to produce transgenic 'Pêra' and 'Valência' sweet orange plants using the D4E1 gene driven by the Arabidopsis thaliana phloem protein (AtPP2) promoter and to quantify transgene expression in different transformation events. Genetic transformation experiments were carried out with epicotyl segments co‑cultivated with Agrobacterium tumefaciens. Six plants from 'Pêra' sweet orange and seven plants from 'Valência' sweet orange were confirmed as different transgenic events by means of the polymerase chain reaction (PCR) and the Southern blot techniques. Transgene expression was quantified using real‑time quantitative PCR. D4E1 gene expression levels vary from 5 up to 50 times among different transformation events.

Effects of biofilm and refrigeration on acerola postharvest conservation

The objective of this work was to compare the effects of four different concentrations of cassava starch film and storage temperature on shelf life and the quality of a genotype of acerola from the Germplasm Active Bank (GAB) of Federal Rural University of Pernambuco, Brazil. Sound orange-reddish acerola fruits were washed with a chlorine solution (100 mg.L-1 active chlorine) and randomly distributed into different lots. The fruits were dipped for 3 min in a cassava starch suspension with concentrations of 1, 2, 3 and 4% (w/v) and the control without coating, stored at 10ºC (85% RH) and 22ºC (85% RH). The total soluble solids (TSS), pH, titratable acidity (TA) and ascorbic acid (AA) were determined at harvest and regular interval during storage. The use of cassava biofilm at 1% on acerola fruits maintained the highest ascorbic acid content and the temperature of 10ºC extended storage life. The fruits coated with 1 and 2% biofilm could be stored for a period up to 15 days at 10ºC, with acceptable quality characteristics.

Residues of carbosulfan and its carbofuran metabolites and 3-hydroxy-carbofuran in oranges

The objectives of this study were to evaluate the residues of the insecticide carbosulfan and its carbofuran metabolites and 3-hydroxy-carbofuran in orange compartments (whole fruit, bagasse and juice) and comparison between the residual levels found in fruits with the maximum residue level and the safety interval established by the Brazilian legislation. Two field experiments were carried out, both with the following treatments: a-check; b-one application of 10 g of carbosulfan . 100 L-1 of water; c-one application with twice the rate applied in treatment b; d-four applications with the same rate applied in treatment b. Samples were taken at (-1), zero, 1, 3, 7, 14, 21 and 28 days after the last or unique application. The quantitative determinations were done by gas chromatography technique, using a nitrogen-phosphorus detector. The carbosulfan metabolism to its carbofuran metabolite was rapid (3 days), being both analytes concentrated in the bagasse (peel + flavedo + albedo). However, the metabolism of carbofuran to 3-hydroxy-carbofuran was of low intensity or this metabolite was quickly dissipated. Carbosulfan residues and its metabolites did not penetrate into the fruit, thus not contaminating the juice. The use of the pesticide was adequate, with respect to fruit consumption, in relation to the Brazilian legislation.

Qualidade dos frutos de tipos de melão,produzidos em ambiente protegido

O interesse pela cultura do melão no Brasil tem aumentado muito nos últimos anos, pelas crescentes exportações e pelo incremento no consumo do mercado interno. O objetivo deste trabalho foi avaliar as características físicas e químicas, assim como a atividade da pectinametilesterase dos frutos de tipos de melão (Cucumis melo L.), produzidos em ambiente protegido, no município de Centenário do Sul-PR. Os tipos estudados foram: Valenciano ('Amarelo-Ouro'), Caipira ('Gaúcho Caipira'), Net Melon ('Net Galia'), Orange ('Orange Melon') e Pele-de-Sapo ('Filipo'), com cinco repetições, utilizando seis frutos por repetição em delineamento inteiramente casualizado. Os frutos do Valenciano e Pele-de-Sapo destacaram-se quanto à massa, com valores 2,02 e 2,07 kg, respectivamente, e formatos alongados, enquanto os demais tipos apresentaram formatos arredondados e massa em torno de 1,4 kg. Os melões Pele-de-Sapo apresentaram espessura da polpa de 43,36 mm, estatisticamente superior à dos frutos Valenciano, com 38,98 mm. A menor espessura de polpa, 24,78 cm, e a maior espessura de casca, 9,74 mm, foram encontradas nos frutos do tipo Caipira que diferiu estatisticamente dos outros tipos. Os valores de pH não se apresentaram estatisticamente diferentes e variaram de 6,24 a 6,48. O Net Melon apresentou polpa com 12,3ºBrix e diferiu estatisticamente do Orange, Valenciano e Pele-de-Sapo, com 11;12; 10,34 e 9,94 ºBrix, respectivamente. O Caipira atingiu 5,06ºBrix, e também o menor conteúdo de acidez, 0,10 g de ac. cítrico.100-1 g de suco, o que inviabiliza sua comercialização. A atividade da pectinametilesterase na polpa dos frutos foi muito baixa, inferior a 0,005 PEu x 10(4) mL-1, nos cinco tipos avaliados. Na região norte do Paraná (Vale do Paranapanema), sob condições de cultivo protegido, os melões Pele-de-Sapo, Net Melon, Orange e Valenciano apresentaram boas características físicas e químicas dos frutos, destacando-se o Net Melon pelo teor de sólidos solúveis totais, acima de 12ºBrix.

Passiflora edulis Sims: the correct taxonomic way to cite the yellow passion fruit (and of others colors)

Passiflora edulis, the passion fruit native from Brazil, has several common names (such as sour passion fruit, yellow passion fruit, black passion fruit, and purple passion fruit), and presents a wide variability with the different rind colors of its fruits, which are very easy to notice. However, in 1932, Otto Degener suggested that the yellow passion fruit had its origin in Australia through breeding, calling it P. edulis forma flavicarpa, and that it could be distinguished by the color of the fruit, the deeper shade of purple of the corona, and the presence of glands on the sepals. These distinctions do not support themselves, for the glands are common to the species (although they may be absent), and the corona has a wide range of colors, regardless of the color of the fruit. A more critical ingredient is the fact that the external coloration of the fruit is a character of complex inheritance and is not dominant, thus displaying a number of intermediate colors, making it difficult to identify the extreme colors. For the correct scientific naming of agricultural plants, the International Code of Botanical Nomenclature must be used in conjunction with the International Code of Nomenclature for Cultivated Plants, with the selections with significant agronomic characteristics recognized and named cultivars. In accordance with the international convention promoted by the UPOV, of which Brazil is a signatory, several colors (light yellow, yellow, orange yellow, pink red, red, red purple, green purple, purple, and dark purple) can be recognized in order to adequately characterize passion fruit cultivars within the species P. edulis. At taxonomic level, Passiflora edulis Sims must be used for any plant and color of sour passion fruits, in combination with a cultivar name for the selected materials.

Use of the heat dissipation method for sap flow measurement in citrus nursery trees1

Sap flow could be used as physiological parameter to assist irrigation of screen house citrus nursery trees by continuous water consumption estimation. Herein we report a first set of results indicating the potential use of the heat dissipation method for sap flow measurement in containerized citrus nursery trees. 'Valencia' sweet orange [Citrus sinensis (L.) Osbeck] budded on 'Rangpur' lime (Citrus limonia Osbeck) was evaluated for 30 days during summer. Heat dissipation probes and thermocouple sensors were constructed with low-cost and easily available materials in order to improve accessibility of the method. Sap flow showed high correlation to air temperature inside the screen house. However, errors due to natural thermal gradient and plant tissue injuries affected measurement precision. Transpiration estimated by sap flow measurement was four times higher than gravimetric measurement. Improved micro-probes, adequate method calibration, and non-toxic insulating materials should be further investigated.

Control of Guignardia citricarpa by Bacillus subtilis and Trichoderma spp.

The ability of isolates of Bacillus subtilis and Trichoderma spp. to control citrus black spot (CBS) was investigated in ´Natal´ sweet orange orchards. The first experiment was conducted during the 2001/2002 season and four isolates of B. subtilis (ACB-AP3, ACB-69, ACB-72 and ACB-77), applied every 28 days, alone or in combination were tested and compared with fungicide treatments. Two other experiments were carried out during the 2002/2003 season, where the same isolates of Bacillus and two isolates of Trichoderma (ACB-14 and ACB-40) were tested being applied every 28 days in the second experiment, and every 15 days in the third experiment. In the first experiment, the treatment with ACB-69 differed statistically from the control, but did not differ from other biological control agents or mixture of Bacillus isolates. In the second experiment, the treatments with ACB-69 and ACB-AP3 resulted in smaller disease index compared with the control treatment. However, this result was not repeated in the third experiment, where the isolates were applied every 15 days. Disease severity was high in both evaluated seasons and the fungicide treatment was the most effective for disease control.

Avaliação genética de seleções e híbridos de limões cravo, volkameriano e rugoso como porta-enxertos para laranjeiras Valência na presença da morte súbita dos citros

Este estudo teve como objetivo realizar a avaliação genética da produção de frutos, eficiência produtiva e altura de laranjeiras Valência (Citrus sinensis) enxertadas em seleções e híbridos dos limões Cravo (C. limonia), Volkameriano (C. volkameriana) e Rugoso (C. jambhiri), em área endêmica para morte súbita dos citros (MSC). Foram avaliados 36 genótipos desses porta-enxertos, representados por cinco plantas cada, avaliados em cinco safras, do terceiro ao sétimo ano após o plantio. Sete dos genótipos avaliados apresentaram plantas com sintomas de MSC até o sétimo ano: Rangpur Otaheite orange 12901 (859), Rangpur Red Lime D.33.30 (866), Limão-Cravo EEL (871), Rangpur Borneo red (874), Citrus kokhai (1649), Limão-Rugoso 58329 (1655) e Limão- Cravo x Swingle B (1695). Para os genótipos que não manifestaram sintomas da doença, foram estimados parâmetros genéticos e fenotípicos e realizada a predição de valores genéticos dos indivíduos, visando à seleção e ao melhoramento genético para as características citadas, empregando-se o método REML/BLUP (máxima verossimilhança restrita/melhor predição linear não viciada). A análise de produção de frutos de cinco safras mostrou acurácia seletiva de 84,59%, tornando-se desnecessária a avaliação de maior número de safras. A seleção dos sete melhores genótipos proporcionou ganhos genéticos de 11,5% na produção de frutos, enquanto a do melhor genótipo conferiu ganho genético de 16,3%. As maiores médias genéticas preditas (>70,0 kg.pl-1) para produção de frutos foram obtidas pelos genótipos Limão-Cravo- Ipanema (1522), Santa- Bárbara-Red- Lime (884), Limão- Cravo- Limeira (863), Limão- Cravo- Taquaritinga (869), Limão- Rugoso- do -Cabo (1643), Rangpur- Rose Lime (868) e Limão- Cravo- da- Califórnia (1467). Já a acurácia seletiva da eficiência produtiva, para quatro colheitas, foi 77,4%. Para este caráter, as maiores médias genéticas (>8,0 kg.m-3) foram dos genótipos Rangpur- Lime x Trifoliata 3810 (1648), Rangpur- Lime x Trifoliata 5320 (1644), Limão- Cravo x Citrange- Carrizo (1524), Citrus pennivesiculata (880), Limão- Cravo x Trifoliata- Swingle A (1707), Rangpur- Rose- Lemon 124684 (864), Rangpur- Red -Lime D33.47 (867) e Limão- Cravo -Ipanema (1522). Dentre os 10 melhores genótipos para produção de frutos e para eficiência produtiva, apenas três são coincidentes: Rangpur- Rose -Lime (868), Citrus pennivesiculata (880) e Limão- Cravo-Ipanema (1522).

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.

Hidrogenionic potential (pH) of the attractant, trap density and control threshold for Ceratitis capitata (DIPTERA: TEPHRITIDAE) on Hamlin oranges in São Paulo central region, Brazil

This study evaluated the effect of initial pH values of 4.5, 6.5 and 8.5 of the attractant (protein bait) Milhocina® and borax (sodium borate) in the field, on the capture of fruit flies in McPhail traps, using 1, 2, 4 and 8 traps per hectare, in order to estimate control thresholds in a Hamlin orange grove in the central region of the state of São Paulo. The most abundant fruit fly species was Ceratitis capitata, comprising almost 99% of the fruit flies captured, of which 80% were females. The largest captures of C. capitata were found in traps baited with Milhocina® and borax at pH 8.5. Captures per trap for the four densities were similar, indicating that the population can be estimated with one trap per hectare in areas with high populations. It was found positive relationships between captures of C. capitata and the number of Hamlin oranges damaged, 2 and 3 weeks after capture. It was obtained equations that correlate captures and damage levels which can be used to estimate control thresholds. The average loss caused in Hamlin orange fruits by C. capitata was 2.5 tons per hectare or 7.5% of production.