Enraizamento de estacas de lichia (Litchi chinensis Sonn.)

Estudou-se os efeitos de auxilias exógenas e ácido bórico, no enraizamento de estacas de lichia (Litchi chinensis Sonn.). As estacas foram uniformizadas, com 25 cm de comprimento e 4 folhas cortadas pela metade. Cerca de 2,5 cm da base das mesmas foi mergulhado nos tratamentos: H2O; Boro 150 µg/ml; IBA 5.000 ppm, IBA 2.000 ppm; IBA 5.000 ppm + Boro 150 µg/ml; IBA 2.000 ppm + Boro 150 µg/ml; NAA 3.000 ppm; NAA 1.500 ppm; NAA 3.000 ppm + Boro 150 µg/ml; NAA 1.500 ppm + Boro 150 µ/g/ml. A estaquia foi realizada no mês de setembro (Hemisfério sul), sendo que as estacas foram colocadas em bandejas de isopor, tendo como substrato vermiculita e mantidas sob nebulização intermitente. Os resultados obtidos permitiram concluir que o IBA 5.000 ppm por 1 minuto foi o tratamento mais efetivo, proporcionando 83,33% de estacas enraizadas em 120 dias, enquanto o tratamento testemunha (H2O), apresentou somente 16,67% de estacas enraizadas.

Poliaminas na embriogênese somática em cenoura (Daucus carota L.)

As poliaminas, putrescina (put), espermidina (spd) e espermina (spm) são amplamente distribuídas na natureza, desempenhando importante papel em vários eventos celulares. Considerando que as poliaminas estão envolvidas no processo embriogênico, torna-se importante conhecer as bases para estas alterações no metabolismo. O objetivo do presente trabalho foi observar os teores de poliaminas durante as diferentes fases da embriogênese somátka da cenoura. As análises foram realizadas, através da modificação do método de Flores e Galston a cada 96 horas durante 50 dias da cultura de células em suspensão. As células foram obtidas a partir de calos e inoculadas em meio de Murashige & Skoog (MS), modificado e acrescido de 0,2 mg/l de 2,4-D e 0,1 mg/l de cinetina durante os primeiros 14 dias da cultura, sendo que no restante do período permaneceram em meio MS desprovido de fiorreguladores. Os teores de poliaminas oscilaram durante todo o processo embriogênico, sendo que a putrescina foi a que apresentou maior acúmulo. Estes resultados sugerem que os níveis de espermidina e espermina são reduzidos em relação a putrescina devido a intensa utilização da spd e spm no processo de divisão e diferenciação celular, enquanto que a put parece ter apenas a função de sintetizar as outras duas amidas.

ISOENZYMATIC POLYMORPHISM and ACTIVITY of PEROXIDASES of COMMON BEAN(Phaseolus vulgaris L.) UNDER SALINE STRESS

Uma das utilizações da técnica de cultura de tecidos para o melhoramento vegetal é a identificação de linhas de células que apresentem tolerância ao estresse salino. Para se estudar os mecanismos bioquímicos envolvidos na expressão genética da tolerância a salinidade, calos oriundos de eixos embrionários de quatro cultivares de feijão (Phaseolus vulgaris L.; cultivares IAC - carioca, IAPAR 14, JALO-EEP 558, BAT - 93), foram cultivados em meio sólido Murashige & Skoog (1962), suplementado com NaCl nas concentrações de 0, 20, 40, 60 e 80 mM. Após 14 dias de incubação, os calos foram coletados e analisados quanto aos padrões isoenzimáticos e de atividade das peroxidases. Os cultivares BAT e IAPAR apresentaram duas zonas de atividade em comum na região anódica e apenas uma zona enzimática específica a cada um deles (migração mais rápida).Possivelmente as duas zonas anódicas intermediárias sejam produtos do mesmo loco enzimático, porém com alelos diferentes, consequentemente diferentes mobilidades eletroforéticas. O cv. JALO apresentou duas zonas anódicas de atividade em comum com os cultivares IAC e IAPAR com uma zona anódica exclusiva de migração mais lenta, a qual apresentou atividade mais intensa de todos os cultivares analisados. Este cultivar revelou ainda uma zona catódica provavelmente dimérica e heterozigota nos indivíduos de todos os tratamentos aplicados. Provavelmente, esta é a mesma zona que ocorre em homozigose com fixação do alelo lento para os indivíduos de todos os tratamentos efetuados nos cultivares BAT e IAPAR. O cv. IAC apresentou duas bandas anódicas em comum com os cv. IAPAR e JALO. Apresentou também a banda anódica mais rápida em comum com o cv. IAPAR e uma banda anódica exclusiva de migração mais lenta. Curiosamente, os indivíduos deste cv. mantidos em meio suplementado com 20 mM de NaCl não apresentaram atividade nas três zonas anódicas mais lentas. Ocorreu no cv. IAC uma única zona de atividade catódica, dimérica e heterozigota para os indivíduos provenientes de todos os tratamentos, composta provavelmente de dois alelos diferentes da zona correspondente ao cv. JALO. Amostras provenientes dos tratamentos 40 e 60 mM de NaCl, desta zona catódica, apresentaram maior atividade enzimática. A análise da atividade da peroxidase no extrato bruto, revelou que os cultivares responderam diferentemente ao aumento da concentração salina no meio de cultura, com aumento pronunciado dessa atividade nos cultivares IAC e JALO.

Multiplicação in vitro DE Ficus carica L.: efeito da cinetina e do ácido giberélico

A cultura da figueira é afetada pelo vírus-do-mosaico e a cultura de tecidos é uma alternativa para se proceder à limpeza clonal. Neste trabalho, objetivou-se estudar o efeito da cinetina e GA3 na multiplicação in vitro da figueira. Segmentos nodais foram inoculados em meio de cultura WPM contendo as seguintes combinações de cinetina (0; 0,5; 1; 2 e 4 mg.L-1) e GA3 (0, 2, 4, 6 e 8 mg.L-1). Avaliaram-se número e comprimento dos brotos, peso da matéria fresca e seca da parte aérea, número de raízes, peso da matéria fresca e seca do sistema radicular e de calos. A utilização de 0,5 mg.L-1 de cinetina promoveu melhor taxa de multiplicação in vitro de Ficus carica. O GA3 reduziu a formação e multiplicação dos brotos e induziu ao estiolamento, à hiperidricidade, clorose e necrose apical das plântulas.

Action of auxins and/or boron, in the process of formation of roots in cuttings of coffee (Coffea-Arabica L CV Mundo-Novo)

The objective of present research was to verify the IBA and/or NAA effects with boron, on rooting of coffee (Coffea arabica L. cv 'Mundo Novo') stem cuttings. Therefore it were used stem cuttings from semi-hardwood orthotropous branches, having two nodes, with aproximately 10 cm of lenght. The cuttings were treateds during 24 hours with IBA and/or NAA plus boron solutions. The treatments' effect was observed trough the following observation, 90 days after planting: rooting percentage and number of cuttings with ''callus''. Trough the resultS obtained, it was concluded that, for obtaining more rooting of cuttings, the best treatment was NAA at 200 ppm plus boron or, the combination of IBA and NAA at 200 ppm plus boron.

Composites PVDF-TrFE/BT used as bioactive membranes for enhancing bone regeneration

In this paper a piezoelectric composite membranes were developed for charge generator to promoter bone regeneration on defects sites. Is known that the osteogenesis process is induced by interactions between biological mechanisms and electrical phenomena. The membranes were prepared by mixing Barium Titanate (BT) powders and PVDF-TrFE (PVDF:TrFE = 60:40 mol%) on dimethylformamide medium. This precursor solution was dried and crystallized at 100degreesC for 12 hours. Composites membranes were obtained by following methods: solvent casting (SC), spincoating (SP), solvent extraction by water addition (WS) and hot pressing (HP).The microstructural analysis performed by SEM showed connectivity type 3-0 and 3-1 with high homogeneity for samples of ceramic volume fraction major than 0.50. Powder agglomerates within the polymer matrix was evidenced were observed for composites with the BT volume fraction major than 40%. The composite of ceramic fraction of 0.55 presented the best values of remanent polarization (similar to33 muC/cm(2)), but the flexibility of these composites with the larger ceramic fraction was significantly affected.For in vivo evaluation PVDF-TrFE/BT 90/10 membranes with 3cm larger were longitudinally implanted under tibiae of male rabbit. After 21 days the animals were sacrificed. By histological analyses were observed neo formed bone with a high mitotic activity. In the interface bone-membrane was evidenced a pronounced callus formation. These results encourage further applications of these membranes in bone-repair process.

Tibial segmental bone defect treated with bone plate and cage filled with either xenogeneic composite or autologous cortical bone graft

Tibia segmental defect healing in sheep were clinically, radiographically and histologically evaluated. Twelve young sheep aged four to five months were divided into two groups, G1 and G2. A 3.5 cm long segmental defect was created in the right tibial diaphysis with maintenance of the periosteum. The bone defects in both groups were stabilized with a bone plate combined with a titanium cage. In G1 the cage was filled with pieces of autologous cortical bone graft. In G2 it was filled with a composite biomaterial which consisted of inorganic bovine bone, demineralized bovine bone, a pool of bovine bone morphogenetic proteins bound to absorbable ultra-thin powdered hydroxyapatiteand bone-derived denaturized collagen. Except for one G1 animal, all of them showed normal limb function 60 days after surgery. Radiographic examination showed initial formation of periosteal callus in both groups at osteo-tomy sites, over the plate or cage 15 days postoperatively. At 60 and 90 days callus remodeling occurred. Histological and morphometric analysis at 90 days after surgery showed that the quantity of implanted materials in G1 and G2 were similar, and the quantity of new bone formation was less (p = 0.0048) and more immature in G1 than G2, occupying 51 +/- 3.46% and 62 +/- 6.26% of the cage space, respectively. These results suggest that the composite biomaterial tested was a good alternative to autologous cartical bone graft in this experimental ovine tibial defect. However, additional evaluation is warranted prior to its clinical usage.

Synthesis Pengo System plates for the treatment of long-bone diaphyseal fractures in dogs

A Brazilian orthopaedic company designed a stainless steel plate called Synthesis Pengo System (S.P.S.), which has one fixed and one changeable extremity. According to the assembly of the changeable extremity, it is possible to obtain dynamization or neutralization of the fracture site. Since the S.P.S. plate was developed for use in human patients, the aim of this study was to evaluate this system in long-bone diaphyseal fractures in dogs. Eight dogs with closed diaphyseal fractures of the femur (n = 1), radius and ulna (n = 5), and tibia (n = 2) were used. Patients were aged seven months to three years and weighed 18 to 31.2 kg. The S.P.S. plate was assembled with one fixed extremity and one changeable extremity in dynamization mode. The trail bar was positioned for synthesis modules with holes for cortical screws. The modules were positioned close to one another in two fractures and for away from the fracture site in the others. The bone healing occurred by external callus. Since motion at the fracture site determines the amount of callus required, the secondary bone healing that was observed in all of the cases indicated less rigid fixation of this system. A potential benefit of this system was a lesser interface contact with the bone since it was only done by trail bar. The major disadvantage was the prominence of the implant. It was possible to conclude that the S.P.S. plate appears to be a suitable method for the treatment of diaphyseal fractures in dogs.

Effect of storage of cuttings on the rooting of different cultivars of cut chrysanthemums (Dendranthema grandiflora Tzvelev) for two different seasons

Chrysanthemums are commercially propagated through cuttings. Pre-rooting storage of cuttings in the dark is a common practice among growers and companies that work and trade with chrysanthemum cuttings. Therefore, the maximum storage period for cuttings and differences in tolerance among cultivars has been investigated. Adventitious roots of cuttings can originate in almost any tissue, including the epidermis, stem cortex and pericycle, ray parenchyma, immature xylem and phloem cells, and pith. The aims of this work were to determine the effect of time of cold storage of cuttings (0, 1, 2, 3, 4, 5 and 6 weeks) on the rooting of four cut chrysanthemum cultivars (Super White, Sheena, Dark Orange Reagan and Town Talk) for two seasons of the year (summer and winter), and also to determine the origin of root formation in chrysanthemum cuttings. The study was carried out as a randomized complete block design with 5 replications for each storage treatment. Each plot was comprised of three cuttings that were examined 14 days after the cutting procedure. During the winter, the roots were studied by scanning electron microscopy (SEM) at the Electron Microscope Laboratory. The following conclusions were made: in winter, cold storage affected the rooting of cuttings, mainly after two weeks of storage for all cultivars. The rooting percentage was lower in the winter and the cuttings could be preserved for a shorter period. The source and growth of roots in chrysanthemum cuttings was found to be endogenous. After three days, callus formed in the pericycle and the first emergence of adventitious roots occurred by the fourth day of rooting. During the summer, cold storage could be up to 4 weeks without any problems.

Effect of benzylaminopurine (BAP) on clonal propagation rate of Curcuma longa L.

Curcuma longa L. is used in many countries for its flavor, and medicinal and cosmetic attributes, as well as for its peculiar starch characteristics. These factors have driven an interest in the in vitro propagation of this species, looking for germplasm bank maintenance, production of disease free plants, genetic variability induction from callus, and as a tool for starch research. However, there are few reports concerning the micropropagation of Curcuma longa. The in vitro propagation rate of this species, cultured under two benzylaminopurine (BAP) concentrations, was the aim of this research.

In vitro propagation of Maytenus ilicifolia (Celastraceae) as potential source for antitumoral and antioxidant quinomethide triterpenes production. A rapid quantitative method for their analysis by reverse-phase high-performance liquid chromatography

Cell culture of Maytenus ilicifolia were established in order to produce and to quantify the antitumoral and antioxidant quinonemethide triterpenes. In vitro calli were induced from leaf explants of native plants and cultured in semi-solid medium under controlled conditions of humidity, temperature and photoperiod. The quinonemethide triterpenes showed maximum accumulation in the logarithmic phase growth of the cell culture. A rapid, sensitive and reliable reverse-phase HPLC method was used for quantitative determination of the antitumoral and antioxidant quinonemethide triterpenes, 22β-hydroxymaytenin and maytenin in callus of Maytenus ilicifolia. Well resolved peaks with good detection response and linearity in the range 1.0 - 100 μg/mL were obtained. This quantitative work was performed by an external standard method.

Avaliação radiográfica, densitométrica e histológica do uso de perfurações ósseas para o estímulo de consolidação de fraturas do terço distal do rádio de cães

To stimulate the bone callus development process from distal third of radius, 24 adult mongrel dogs used were from both sexes. These dogs were separated in two experimental groups of 12 animals each, named control and treated, divided in 4 moments (M1=15 days; M2=30 days; M3=45 days; M4=60 days), who underwent were performed surgical fractures. In treated group, it was performed bone perforations on proximal and distal edges, craniolateral and mediolateral to the fracture site. At the end of each moment, control and treated animals were evaluated by radiography, histology, and bone mineral densitometry (BMD) was determined on fracture site. According to the radiographic data of treated dogs, it was verified on days 15 and 30 more intense bone regeneration than control group. During M3 and M4, it wasn't detected any difference in bone reparation process betweencontrol and treated groups. In densitometric study, BMD values were greater in treated animals than in control dogs. Histological studies revealed at 15 and 30 days chondrocyte hyperplasia and initial endochondral ossification on drilled limbs; control group showed sustainment connective tissue and initial chondrocyte hiperplasia. At M3 and M4 of the treated group, were verified development and remodeling of periosteal callus in more advanced phases when comparing with limbs from control group. It can be concluded that using perforations enhances blood flow supply and activation of osteogenic cells on fracture site, stimulating the beginning of fracture consolidation process.

Micropropagación de Pothomorphe umbellata (L.) Miq. vía organogénesis directa

The present research was aimed at standardizing the protocol of seed disinfection, seed germination and organogenesis via callus of Pothomorphe umbellata (L.) Miq.The germinated seeds were inoculated in different concentrations of BAP (benzylamine purine) and NAA (naphthalene acetic acid) in order to stimulate the callus induction. After 60 days of culture, the calluses with some shoots were taken to an organogenesis medium (GA3 0.1 mg.L -1, BAP 0.5 mg.L-1) during 40 days, to be transferred later to a development medium. Finally, the plantules were acclimatized, presenting a good index of survival.

Carotenóides totais em Pothomorphe umbellata (L.) Miq. cultivadas in vitro

The aim of the present work was to compare the content of carotenoids between callus and regenerated plants of Pothomorphe umbellate. Germinated seeds (40 days old) were inoculated in different concentrations and combinations of BAP (benzylaminopurine) and NAA (naphthalene acetic acid) in order to stimulate the callus' production. After 60 days of culture, the callus containing some shoots were transferred to organogenesis medium (GA 3 0.1 mg L -1, BAP 0.5 mg L -1) for 40 days. Next, they were subcultivated in a medium for seedling growth (without regulators) for 40 days. Callus (collected after 60 days) and seedlings (collected after 140 days) were frozen in liquid nitrogen and kept under -80°C for future carotenoids' analysis. The highest concentration of carotenoids was found in plants cultivated in medium without regulators. The callus did not showed difference concerning the culture medium; however, they presented lower content of carotenoids in relation to plants.

Growth regulators for in vitro propagation of Tillandsia gardneri (Bromeliaceae)

Tillandsia gardneri Lindi, is a herbaceous perennial with ornamental value. However, in Brazil there is no report about this species' cultivation on a commercial scale. The low multiplication rate of T. gardneri (in average one offshoot/plant/year) leads to illegal over-collection in the wild to meet commercial demands. The development of protocols for in vitro propagation of T. gardneri may be useful for increasing multiplication rate, producing enough plants to supply the ornamental market and also to reduce the pressure over plants collection in the wild. The present study evaluated the effect of growth regulators (6-benzylaminopurine-BA alone or in combinations with naphthaleneacetic acid-NAA) on shoots development from seedlings pre-established in vitro, from seed germination on 1/4 MS medium without growth regulators. Seedlings (with about 1.0 cm long) were re-cultured to solid 1/2 MS media supplemented with growth regulators. After 30 days on the induction medium seedlings were re-cultured to MS basal medium. The experiment was conducted in a complete randomized design with four replications and ten treatments: control (free of growth regulators), BA (0.5, 1.0 and 2.0 mg/L), BA (0.5 mg/L) + ANA (0.1 mg/L), BA (1.0 mg/L) + ANA (0.1 mg/L), BA (1.0 mg/L) + ANA (0.5 mg/L), BA (2.0 mg/L) + ANA (0.1 mg/L), BA (2.0 mg/L) + ANA (0.5 mg/L), and BA (2.0 mg/L) + ANA (1.0 mg/L). The outgrowth of shoots did not occur on medium devoid of growth regulators (control). Regression analysis for some evaluated parameters, such as percentage of seedlings responsive to shoot formation and number of shoots/seedling, and regulators concentrations (BA or ANA) were significant, allowing the establishment of the growth regulators concentration for obtaining the best multiplication rate. Some seedlings maintained in media with ANA (0.5 or 1.0 mg/L) were completely converted into callus masses that turned dark brown leading to seedlings death.