Enraizamento de plantas cultivadas in vitro

The adventitious rooting process of in vitro cultured plantlets is a technique that has been employed for the vegetative propagation of a significant number of native and exotic species. Many factors are associated with the rooting stage influencing positive and/or negatively the establishment of micropropagation protocols. The objective of this work was a literature review of the main inherent factors concerning in vitro rooting process including the correlation among others the endogenous and exogenous auxins levels, juvenility, genotype, mineral nutrition, culture medium conditions, addition of growth regulators and other substances as phenolic compounds and active coal besides growth environmental conditions of in vitro cultures. Although the complete elucidation of all processes involved with rooting of in vitro cultured plants has not been achieved so far, a comprehensive study of the main factors that interfere on rooting is fundamental for the establishment of new researches that might contribute for the rooting of economically important plants.

Meio de cultura e tipo de explante no estabelecimento in vitro de espécies de maracujazeiro

Brazil is one of the main centers of genetic variability dispersion of the Passiflora genera. Its self incompatibility as well as disease incidence in its leaves and root system and, deforestation and monocultivation, promote loss of genetic material. Considering the risk of genetic erosion, the conservation of the variability in germplasm banks, which is of great interest in plant breeding, is necessary. Studies regarding the type of expiant and concentration of the culture media are necessary in order to determine protocols of establishment and in vitro conservation of passion-fruit germplasm. The objective of the present work was to evaluate the influence of the salt and nutrient concentration in the MS culture medium and types of expiants in the establishment and growth of the Passion fruit species: Passiflora giberti N. E.Brown, P. edulis Sims and P. laurifolia L. Each Passiflora species presented its own characteristics regarding in vitro development. The complete MS medium and nodal segments the second axilliary bud promoted better development of the genotypes studied.

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.

Seed and seedling surface-sterilization for in vitro culture of tillandsia gardneri (Bromeliaceae)

Tillandsia gardneri is a bromeliad with ornamental value and a wide geographical distribution over Brazil. However, due to habitat loss and illegal overcollection in the wild it is included as a vulnerable species in the official list of endangered plants of the State of Rio Grande do Sul, Brazil. The development of a protocol for T. gardneri seed propagation in vitro may be useful for reintroducing plants in their natural habitats, and for germplasm conservation. A difficult problem encountered during the establishment of an in vitro culture is explants disinfection, especially when working with endangered species, from which explant availability is restricted. Thus, the establishment of a sterilization protocol is crucial for the initiation and success of a micropropagation system for T. gardneri. The objective of this study was to evaluate the effect of sodium hypochlorite concentration and exposure time in seed and seedling surface disinfection, tissue sensitivity and development. Sodium hypochlorite solutions (10 or 20%/5, 10 or 15 min; 25%/5 or 10 min; and 50%/5 min) were effective in eliminating seed superficial contaminants. There was no significant difference among the effective sterilization treatments in relation to seed germination (%), and seedling length and number of leaves, after 120 days in vitro. Also, no damage to seed and seedling tissues were observed. Surface sterilization of seedlings, for initiation of an in vitro culture, required higher concentrations of sodium hypochlorite (25%/15 min; 20 or 50%/5, 10 or 15 min; and 40%/5 and 10 min) for controlling fungal and yeast contamination, compared to seed sterilization. No significant differences among these treatments were found in relation to seedling length and number of leaves, after 60 days in vitro.

Effects of ascorbic acid on in vitro culture of bovine preantral follicles

The objective of this study was to evaluate the effects of adding ascorbic acid to the media for in vitro culture of cattle ovarian fragments and to determine their effects on growth activation and viability of early-stage follicles. The ovarian cortex was divided into small fragments; one fragment was immediately fixed (control) and the other fragments were cultured in minimum essential medium (MEM) supplemented or not with various doses of ascorbic acid. Ovarian tissue was processed for histology, transmission electron microscopy (TEM) and immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA). Compared with control fragments, the percentage of primordial follicles was reduced (p < 0.05) and the percentage of growing follicles had increased (p < 0.05) in cultured cortical fragments, independent of the tested medium or incubation time. Furthermore, compared with control tissue, culture of ovarian cortex for 8 days reduced the percentages of healthy, viable follicles (p < 0.05), but not when cultures were supplemented with 25, 50 or 100 μg/ml of ascorbic acid. Ultrastructural and immunohistochemical analysis of 8 day cultured ovarian cortical fragments, however, showed the integrity and viability of follicles only when fragments were cultured in presence of 50 μg/ml of ascorbic acid. In conclusion, this study demonstrated that addition of ascorbic acid to MEM at a concentration of 50 μg/ml not only stimulates the activation of 8 day in vitro cultured cattle primordial follicles and subsequent growth of activated follicles, but also safeguards the viability of these early-stage follicles. © 2012 Copyright Cambridge University Press.

Supplementation with the histone deacetylase inhibitor trichostatin A during in vitro culture of bovine embryos

Summary Trichostatin A (TSA) is a histone deacetylase inhibitor that induces histone hyperacetylation and increases gene expression levels. The aim of the present study was to establish a suitable condition for the use of TSA in in vitro cultures of bovine embryos, and to determine whether TSA would increase blastocyst rates by improvement of chromatin remodelling during embryonic genome activation and by increasing the expression of crucial genes during early development. To test this hypothesis, 8-cell embryos were exposed to four concentrations of TSA for different periods of time to establish adequate protocols. In a second experiment, three experimental groups were selected for the evaluation of embryo quality based on the following parameters: apoptosis, total cell number and blastocyst hatching. TSA promoted embryonic arrest and degeneration at concentrations of 15, 25 and 50 nM. All treated groups presented lower blastocyst rates. Exposure of embryos to 5 nM for 144 h and to 15 nM for 48 h decreased blastocyst hatching. However, the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay revealed similar apoptosis rates and total cell numbers in all groups studied. Although, in the present study, TSA treatment did not improve the parameters studied, the results provided background information on TSA supplementation during in vitro culture of bovine embryos and showed that embryo quality was apparently not affected, despite a decrease in blastocyst rate after exposure to TSA. © Cambridge University Press 2011.

Pulp fruit added to culture medium for in vitro orchid development

As an additive in in vitro culture media, fruits have a great potential for facilitating economical orchid production because of lower technology requirements and the ease of obtaining raw materials to formulate culture media. We studied the in vitro growth of Cattleya bicolor Lindl. grown in a simplified culture medium supplemented with different kinds of fruit pulp. The experimental design was completely randomised, with eight seedlings per replication and ten replications per treatment, for a total of 80 seedlings per treatment. The culture medium was made using 150 g L -1 of pulp (without peel or seed) from the following fruits: ripe Santa Cruz tomatoes (Solanum lycopersicum L.), dwarf bananas (Musa cavendishii L.) of intermediate ripeness, light green chayote (Sechium edule (Jacq.) Sw), ripe papaya (Carica papaya L.) or green coconut (Cocos nucifera L.).The treatment control was MS 50 %. The treatments and the control were kept in a growth chamber for seven months before evaluating seedling survival percentage, shoot height, number of leaves, rooting percentage, root number, root length and dry masses of shoot and roots. The highest percentages of seedling survival were obtained using MS 50 %, banana and coconut medium. The seedling survival and rooting percentages illustrate that it is possible to emphasise the culture medium MS 50% and the culture medium supplemented with coconut on the most traditional culture medium with banana or tomato pulp. For the in vitro development of Cattleya bicolor Lindl., a simplified culture medium supplemented with coconut pulp is the most suitable for use as an alternative to MS 50%. A simplified culture medium supplemented with papaya pulp is not recommended for the in vitro development of Cattleya bicolor Lindl.

Avaliação in vitro de diferentes métodos de análises de fungitoxicidade de óleos essenciais

Environmental problems caused by synthetic fungicides have increased the search for alternative methods of control of plant diseases. The objective was to evaluate the effect of essential oil of citronella grass, on the fungus Rhizoctonia solani, in different methods of in vitro fungitoxicity. We used a randomized design in a factorial design with four replications, where the factors were composed of four methods for assessing the in vitro fungitoxicity of the essential oil of citronella grass (essential oil diluted in Tween 80 (0.5%) and embedded in the culture medium PDA (potato dextrose agar) still melting, essential oil diluted in Tween 80 (0.5%) and distributed on the surface of the PDA; oil essential diluted in Tween 80 (0.5%) and distributed on filter paper attached to the inner surface of the lid of the Petri dish, pure essential oil and distributed on the surface of the culture medium, and control) and five evaluation periods (2, 4, 6, 8 and 10 days of incubation). Was used 0.25μL mL-1 of citronella oil in all treatments. Of the treatments evaluated the use of pure oil distributed on the surface of the culture medium was more effective in reducing the mycelial diameter in all evaluations. In this method the rate of mycelial growth was 9,02 mm day-1, reaching in last evaluation 79,77 mm.

Seiva Sangue de Dragão (Croton lechleri) como meio de estocagem de dentes avulsionados: estudo in vitro da viabilidade celular

Pós-graduação em Odontologia - FOA

Reguladores vegetais no desenvolvimento in vitro de bromélia (Aechmea blanchetiana)

Pós-graduação em Agronomia (Horticultura) - FCA

Avaliação do efeito anti-inflamatório da silibinina sobre monócitos de sangue periférico de indivíduos saudáveis, infectados in vitro com cepa virulenta de Paracoccidioides brasiliensis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Reguladores vegetais e poliaminas na organogênese in vitro de Piper hispidinervium Candolle de Candolle: análises biométricas e bioquímicas

Pós-graduação em Ciências Biológicas (Botânica) - IBB

Caracterização morfológica, química e conservação in vitro de Pfaffia glomerata(Sprengel)Pedersen

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Potencial morfogenético e carotenóides em tecidos cultivados in vitro de Pothomorphe umbellata L

Pós-graduação em Agronomia (Horticultura) - FCA

Perfil transcricional de Bradyrhizobium elkanii SEMIA 587 in vitro e em simbiose com soja (Glycine max L. Merrill) através de microarranjo de DNA

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)