Caracterização fisiológica de maçã Royal gala minimamente processada submetida a diferentes tratamentos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Novel insights into the genomic basis of citrus canker based on the genome sequences of two strains of Xanthomonas fuscans subsp aurantifolii

Background: Citrus canker is a disease that has severe economic impact on the citrus industry worldwide. There are three types of canker, called A, B, and C. The three types have different phenotypes and affect different citrus species. The causative agent for type A is Xanthomonas citri subsp. citri, whose genome sequence was made available in 2002. Xanthomonas fuscans subsp. aurantifolii strain B causes canker B and Xanthomonas fuscans subsp. aurantifolii strain C causes canker C. Results: We have sequenced the genomes of strains B and C to draft status. We have compared their genomic content to X. citri subsp. citri and to other Xanthomonas genomes, with special emphasis on type III secreted effector repertoires. In addition to pthA, already known to be present in all three citrus canker strains, two additional effector genes, xopE3 and xopAI, are also present in all three strains and are both located on the same putative genomic island. These two effector genes, along with one other effector-like gene in the same region, are thus good candidates for being pathogenicity factors on citrus. Numerous gene content differences also exist between the three cankers strains, which can be correlated with their different virulence and host range. Particular attention was placed on the analysis of genes involved in biofilm formation and quorum sensing, type IV secretion, flagellum synthesis and motility, lipopolysacharide synthesis, and on the gene xacPNP, which codes for a natriuretic protein. Conclusion: We have uncovered numerous commonalities and differences in gene content between the genomes of the pathogenic agents causing citrus canker A, B, and C and other Xanthomonas genomes. Molecular genetics can now be employed to determine the role of these genes in plant-microbe interactions. The gained knowledge will be instrumental for improving citrus canker control.

A Component of the Xanthomonadaceae Type IV Secretion System Combines a VirB7 Motif with a N0 Domain Found in Outer Membrane Transport Proteins

Type IV secretion systems (T4SS) are used by Gram-negative bacteria to translocate protein and DNA substrates across the cell envelope and into target cells. Translocation across the outer membrane is achieved via a ringed tetradecameric outer membrane complex made up of a small VirB7 lipoprotein (normally 30 to 45 residues in the mature form) and the C-terminal domains of the VirB9 and VirB10 subunits. Several species from the genera of Xanthomonas phytopathogens possess an uncharacterized type IV secretion system with some distinguishing features, one of which is an unusually large VirB7 subunit (118 residues in the mature form). Here, we report the NMR and 1.0 angstrom X-ray structures of the VirB7 subunit from Xanthomonas citri subsp. citri (VirB7(XAC2622)) and its interaction with VirB9. NMR solution studies show that residues 27-41 of the disordered flexible N-terminal region of VirB7(XAC2622) interact specifically with the VirB9 C-terminal domain, resulting in a significant reduction in the conformational freedom of both regions. VirB7(XAC2622) has a unique C-terminal domain whose topology is strikingly similar to that of N0 domains found in proteins from different systems involved in transport across the bacterial outer membrane. We show that VirB7(XAC2622) oligomerizes through interactions involving conserved residues in the N0 domain and residues 42-49 within the flexible N-terminal region and that these homotropic interactions can persist in the presence of heterotropic interactions with VirB9. Finally, we propose that VirB(7XAC2622) oligomerization is compatible with the core complex structure in a manner such that the N0 domains form an extra layer on the perimeter of the tetradecameric ring.

Genetic transformation of Citrus sinensis cv. Hamlin with hrpN gene from Erwinia amylovora and evaluation of the transgenic lines for resistance to citrus canker

Transgenic Citrus sinensis (L.) Osb. cv. Hamlin plants expressing the hrpN gene were obtained by Agrobacterium tumefaciens (Smith and Towns) Conn-mediated transformation. hrpN encodes a harpin protein, which elicits the hypersensitive response and systemic acquired resistance in plants. The gene construct consisted of gst1, a pathogen-inducible promoter, a signal peptide for protein secretion to the apoplast, the selection genes nptI1 or aacC1 and the Nos terminator. The function of gst1 in citrus was evaluated in transgenic C. sinensis cv. Valencia harboring the reporter gene uidA (gus) driven by this promoter. Histochemical analysis for gus revealed that gst1 is activated in citrus leaves by both wounding and inoculation with Xanthomonas axonopodis Starr and Garces pv. citri (Hasse) Vauterin et al. Genetic transformation was confirmed by Southern blot hybridization in eight cv. Hamlin acclimatized plants. RT-PCR confirmed hrpN gene expression in seven cv. Hamlin transgenic lines before pathogen inoculation. Some hrpN transgenic lines showed severe leaf curling and abnormal growth. Six hrpN transgenic lines were propagated and evaluated for susceptibility to X axonopodis pv. citri. RT-PCR confirmed gene expression in all six hrpN transgenic lines after pathogen inoculation. Several of the hrpN transgenic lines showed reduction in susceptibility to citrus canker as compared with non-transgenic plants. One hrpN transgenic line exhibited normal vegetative development and displayed very high resistance to the pathogen, estimated as up to 79% reduction in disease severity. This is the first report of genetic transformation of citrus using a pathogen-inducible promoter and the hrpN gene. Further evaluations of the transgenic plants under field conditions are planned. Nevertheless, the evidence to date suggests that the hrpN gene reduces the susceptibility of citrus plants to the canker disease. (C) 2009 Elsevier B.V. All rights reserved.

Aceitação e preferência de frutos para oviposição em duas populações de Ceratitis capitata (Diptera, Tephritidae)

The influence of four host fruits, orange (Citrus sinensis L.), papaya (Carica papaya L.), mango (Mangifera indica L.) and apple (Malus domestica Borkh) on oviposition behavior of Ceratitis capitata (Wiedemann, 1824) was evaluated. Experiments were carried out on two C. capitata laboratory-reared populations: one with artificial diet for 10 years with periodic introduction of wild flies and other reared with artificial diet for 20 years without wild flies introduction. In acceptance experiments two pieces of a single fruit type were exposed to a group of 10 females; after 48 hours, these were changed by two new pieces of the same fruit type, and, in the end of the fourth day, the experiments were concluded. In preference experiments, two pieces of different fruits were offered to the females. These experiments were driven under the same conditions of the acceptance ones. Acceptance and preference experiments for both populations showed the following choice hierarchy: papaya > mango > orange > apple. The acceptance experiments showed no differences in the number of eggs/female/day laid in the first 48 hours and in the last 48 hours of the experiments. The amount of eggs in the two pieces of fruits offered to the females was similar. In spite of the two populations present similar behavior in relation to host hierarchy, the number of laid eggs was different, being larger for the population reared without wild flies introduction.

Copper acquisition by the SenC protein regulates aerobic respiration in Pseudomonas aeruginosa PAO1.

Aerobic respiration of Pseudomonas aeruginosa involves four terminal oxidases belonging to the heme-copper family (that is, three cytochrome c oxidases and one quinol oxidase) plus one copper-independent, cyanide-insensitive quinol oxidase (CIO). The PA0114 gene encoding an SCO1/SenC-type protein, which is known to be important for copper delivery to cytochrome c in yeast, Rhodobacter spp. and Agrobacterium tumefaciens, was found to be important for copper acquisition and aerobic respiration in P. aeruginosa. A PA0114 (senC) mutant grew poorly in low-copper media and had low cytochrome cbb(3)-type oxidase activity, but expressed CIO at increased levels, by comparison with the wild-type PAO1. Addition of copper reversed these phenotypes, suggesting that periplasmic copper capture by the SenC protein helps P. aeruginosa to adapt to copper deprivation.

Efeito de auxinas sintéticas no enraizamento in vitro da macieira

Brotações de macieira (Malus domestica, Borkh), cv. Fred Hough, oriundas do processo de multiplicação in vitro, foram inoculadas em meio MS e MS/2, testando-se os reguladores de crescimento: ácido indol-3-acético (AIA); ácido indolbutírico (AIB) e ácido naftaleno acético (ANA), nas concentrações de 0, 1, 3 e 5 miM com o objetivo de observar o efeito dessas auxinas sobre o enraizamento da cultivar. Foram acrescentadas aos meios as vitaminas MS mio-inositol (100 mg/L) e sacarose (30 g/L) em meio de ágar (6 g/L). O pH do meio foi ajustado para 5,8 e a cultura foi incubada a 25 ± 2º C e 16 horas de fotoperíodo a 2.000 lux, permanecendo por 30 dias. Os tratamentos foram repetidos cinco vezes e cada repetição constou de cinco explantes inoculados em frasco de 250 mL contendo 40 mL do meio. O meio MS/2 em todas as concentrações testadas foi melhor que o MS. O ANA e o AIB, ambos na concentração de 3 miM, em meio MS/2, tiveram comportamento semelhante na porcentagem de enraizamento e no número de raízes produzidas; no entanto, o ANA provocou efeitos indesejáveis na qualidade destas, havendo formação de calo na base das brotações e raízes grossas. O AIA obteve melhor resposta nas altas concentrações, mas não foi melhor que o AIB e ANA.

Efeito de diferentes volumes de calda no raleio químico de frutos da macieira 'Gala'

O objetivo deste trabalho foi comparar a eficiência de diferentes volumes de calda de raleantes químicos em plantas de macieira (Malus domestica Borkh), cultivar Gala. Foram utilizados o ácido naftalenoacético (ANA) (15 g/ha de i.a.) e o carbaryl (1naftilNmetilcarbamato) (1.000 g/ha de i.a.) aplicando-se 16, 300, 430, 950, 1.300 e 1.900 L/ha de calda. O ANA foi aplicado cinco dias após a plena floração (DAPF), e o carbaryl, aos 15 DAPF. O volume de calda não parece ter sidoo fator mais importante para a eficiência do raleio: os tratamentos utilizados provocaram respostas semelhantes nas plantas. O peso e a porcentagem de frutas >66 mm, em todos os tratamentos, com exceção de 430 L/ha, foram maiores do que os da testemunha, mas não diferiram entre si.

Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange

The development and optimization of efficient transformation protocols is essential in new citrus breeding programs, not only for rootstock, but also for scion improvement. Transgenic 'Hamlin' sweet orange (Citrus sinensis (L.) Osbeck) plants were obtained by Agrobacterium tumefaciens-mediated transformation of epicotyl segments collected from seedlings germinated in vitro. Factors influencing genetic transformation efficiency were evaluated including seedling incubation conditions, time of inoculation with Agrobacterium and co-culture conditions. Epicotyl segments were adequate explants for transformation, regenerating plants by direct organogenesis. Higher percentage of transformation was obtained with explants collected from seedlings germinated in darkness, transferred to 16 hours photoperiod for 2-3 weeks, and inoculated with Agrobacterium for 15-45 min. The best co-culture condition was the incubation of the explants in darkness, for three days in culture medium supplemented with 100 muM of acetosyringone. Genetic transformation was confirmed by performing beta-glucoronidase (GUS) assays and, subsequently, by PCR amplification for the nptII and GUS genes.

Thidiazuron effects on shoot growth, return bloom, fruit set and nutrition of apples

Apple (Malus domestica Borkh.) trees, cultivars Gala and Fuji, were sprayed at full bloom with thidiazuron (TDZ) at doses of 0, 5, 10, or 20 g ha-1 of a.i. in order to evaluate its effects on plant growth and development, return bloom, fruit set, nutrition, and fruit yield. Fruit set increased with TDZ dose in 'Gala' but not in 'Fuji'. TDZ did not affect fruit yield in any cultivar. In 'Gala', the return bloom was reduced in about 50% at TDZ doses of 10 or 20 g ha-1. TDZ increased shoot growth in both cultivars. In leaves, TDZ decreased the concentration of Ca and Mg in 'Gala' and of Mg in 'Fuji', but did not affect the chlorophyll content, leaf area, length, width, and dry mass per cm² regardless of cultivar. In fruits, the effect of TDZ varied according to the portion evaluated. Highest doses of TDZ decreased the concentration of Ca and K in 'Gala' and of K in 'Fuji' in the entire fruits (flesh + skin); in the skin, highest doses of TDZ reduced the levels of N, Ca, and Mg in both cultivars, in addition to the level of K in 'Fuji'.

Deposição de agrotóxicos pulverizados na cultura da maçã

O objetivo deste trabalho foi avaliar a distribuição da deposição de agrotóxicos pulverizados em duas cultivares de maçã (Malus domestica Borkh). Uma calda com traçador Rodamina B foi pulverizada, com equipamento do tipo carreta com cortina de ar, com dois tipos de calibrações. Em uma calibração foram utilizados bicos cones JA-1, JA-2 e JA-3 e na outra, bicos leque AVI 110 ISO-04 e bicos JA-1. Não houve diferenças entre as calibrações, mas a deposição nas plantas foi decrescente da região apical para a basal. Nas calibrações testadas, 25% do traçador aplicado perdeu-se no solo.

PERDAS QUALITATIVAS DE MAÇÃS CV. GALA EM ARMAZENAMENTO REFRIGERADO

O presente trabalho foi desenvolvido com o objetivo de dimensionar as perdas de qualidade de maçãs cv. Gala durante o armazenamento refrigerado. Para determinar estas perdas, foi feito um acompanhamento durante três anos consecutivos, com frutos de três tamanhos: calibre em torno de 100 frutos/caixa de 18kg, calibre 130 e calibre acima de 200. Quinzenalmente, durante os primeiros 90 dias de armazenagem no primeiro ano, 144 dias no segundo ano e 211 dias no terceiro ano, amostras de 20 a 30 maçãs foram avaliadas para perda de peso e parâmetros qualitativos como a firmeza de polpa, os sólidos solúveis totais (SST) e acidez titulável. As variáveis que apresentaram as melhores correlações com o período de armazenagem e, portanto, podem auxiliar na estimativa de possíveis perdas quantitativas durante o armazenamento em ar-refrigerado foram a firmeza de polpa e a acidez titulável. As maiores perdas de qualidade, com exceção da firmeza de polpa, foram observadas em frutos dos tamanhos menores.

Effect of aminoethoxyvinilglycine (AVG) on preharvest fruit drop and maturity of apples

Apple trees cultivars Gala and Fuji were sprayed four weeks before commercial harvest with aminoethoxyvinilglycine (AVG), at doses of 0, 125, or 250 mg L-1, and assessed for preharvest fruit drop, fruit growth, and maturation on tree. In 'Gala', 64 days after AVG spraying, fruit drop for control treatment was 85%, and AVG (at 125 and 250 mg L-1) reduced it to 10%. In 'Fuji', 64 days after AVG spraying, fruit drop for control was 6%, while treatments with AVG (at 125 and 250 mg L-1) increased fruit drop to 10%. AVG was a powerful retardant of fruit maturation for 'Gala' but not for 'Fuji'. In 'Gala', the most affected attribute was the skin background color, followed, in decreasing order, by soluble solids content, the starch index, skin red color, the flesh firmness, and titratable acidity. In 'Gala', only flesh firmness retention was improved by increasing AVG dose from 125 mg L-1 to 250 mg L-1. The AVG at 250 mg L-1 inhibited "Gala" late fruit growth but not 'Fuji'.

Effect of preharvest spraying with thidiazuron on fruit quality and maturity of apples

Apple trees, cultivars Gala and Fuji, were sprayed at full bloom with thidiazuron (TDZ) at the doses of 0, 5, 10, or 20 g (a.i.) ha-1 and fruit were assessed for quality and maturity. In both cultivars, the increase of TDZ dose had detrimental effects on fruit quality, causing a reduction of fruit red surface and an increase of percentage of fruit that was asymmetrical and with calyx-end rot. TDZ caused an increment of calyx-end aperture that might have increased calyx-end rot. TDZ increased density and the length : diameter (L : D) ratio of the fruit. The number of viable seeds was not affected by TDZ. TDZ resulted in a substantial delay of fruit maturity in 'Gala' but not in 'Fuji'. TDZ also reduced soluble solids content (SSC) in 'Gala' and reduced titratable acidity in both cultivars.

Estudos histológicos preliminares da microenxertia de plantas micropropagadas de macieira

A fruticultura moderna necessita implantar tecnologias que possibilitem a produção de frutos de alta qualidade, com custos cada vez menores. A micropropagação associada à microenxertia possibilita altas taxas de multiplicação de plantas com alta qualidade fitossanitária, além de possibilitar a realização de estudos sobre compatibilidade de enxertia em diferentes clones. O presente trabalho tem como objetivo estudar o processo de soldadura entre genótipos de macieira (Malus domestica. Borkh) multiplicadas in vitro após a microenxertia. Esta técnica foi realizada em fenda simples, sob condições assépticas. Os estudos histológicos foram realizados através de cortes longitudinais seriados de segmentos de 8 mm do ponto de enxertia. O processo de soldadura foi caracterizado pelo desenvolvimento de tecido meristemático, originando células parenquimáticas na interface do microenxerto, com a proliferação do tecido cambial da cultivar copa. Isso possibilita a ligação do sistema vascular da copa com o do porta-enxerto, resultando na sobrevivência do microenxerto.