Cytotoxic effects of the essential oil from leaves of Casearia sylvestris Sw. (Salicaceae) and its nanoemulsion on A549 tumor cell line

Extracts from leaves of C. sylvestris have cytotoxic effect in different tumor cell lines, possibly due to clerodane type diterpenes (casearins). On the other hand, there are few studies related to the antitumor activity of the essential oils from this species. This work evaluated for the first time the cytotoxicity effects of the pure essential oil and its nanoemulsion against A549 tumor cell line (human lung carcinoma). The essential oil was obtained from fresh leaves by hydrodistillation in a Clevenger-type apparatus and analyzed by GC/MS and GC/FID. Cytotoxicity evaluation was performed using the WST-1 test. The chemical analysis of the essential oil revealed a volatile fraction composed mainly of non-oxygenated sesquiterpenes (72.1%). The essential oil and its nanoemulsion exhibited cytotoxic activity against A549 tumor cells with EC50 of 4.0 μg/mL and EC50 of 1.0 μg/mL, respectively. Both samples displayed a dose dependent pattern (r = -0.79, p = 0.03) as determined by linear regression test

Anatomical aspects, chemical analysis and cytotoxic effect of the essential oil from leaves of Casearia arborea (Salicaceae)

The genus Casearia (Salicaceae) is found in sub-tropical and tropical regions of the world and comprises about 200 species. In Brazil, there are about 48 species and 12 are registered in the State of Rio de Janeiro; including Casearia arborea (Rich.) Urb. Essential oil was obtained from the fresh leaves by hydrodistillation and analyzed by GC-MS and GC-FID. The cytotoxic effect was determined by WST-1 assay. Chemical analysis of the essential oil revealed a very diversified (n = 37 compounds) volatile fraction composed mainly of non-oxygenated sesquiterpenes (90.2%). These sesquiterpenes included byciclogermacrene (18.7%), germacrene D (12.1%) and α- humulene (11.5%). In addition, the essential oil demonstrated cytotoxic effects against A549 tumor cells in the concentration of 4 μg/mL (EC50) (p < 0.05)

Estabilidad genética entre clones de berenjena in vitro inducidos por diferentes reguladores de crecimiento de plantas

Many factors may influence the genetic stability of plant in vitro clones, among which the genotype and the regenerative process induced by plant growth regulators. The resulting somaclonal variations may be useful for breeding projects, but may be detrimental to germplasm conservation. The objective of this work was to evaluate the genetic stability of Solanum melongena cv. Florida Market clones, obtained in response to different plant growth regulators. For the production of clones, leaf explants were used from commercial seed germinated plants. The explants were inoculated in Murashige and Skoog medium supplemented with different plant growth regulators at pre­defined concentrations. The DNA was extracted by the CTAB method from leaves of complete plants obtained by somatic embryogenesis induced by naphthaleneacetic acid (NAA) or indirect organogenesis induced by benzylaminopurine (BAP) or thidiazuron (TDZ). For the RAPD, 117 DNA samples were amplified by ten decamer primers and 49 specific bands were selected among the products for the comparative study. A total of 5733 fragments were obtained, with a rate of 5.37% polymorphism. NAA did not generate polymorphism and the BAP was responsible for the highest rate obtained (14.28%). Two RAPD primers were identified as markers for monitoring the genetic stability of eggplant. The polymorphic pattern was observed only in clones originating from indirect organogenesis. These results indicate the usefulness of a monitoring protocol for studies using in vitro cloned eggplant.Muchos factores pueden influir en la estabilidad genética de los clones de plantas in vitro, entre los que se encuentran el genotipo y el proceso regenerativo inducido por los reguladores del crecimiento. Por lo tanto, las variaciones somaclonales resultantes del cultivo pueden ser útiles para proyectos de mejoramiento genético, pero pueden ser perjudiciales para la conservación de germoplasma. El objetivo de este estudio fue evaluar la estabilidad genética de Solanum melongena cv. Florida Market, obtenida en respuesta a diferentes reguladores del crecimiento. Para la producción de clones se utilizaron explantes de hojas provenientes de plantas obtenidas de semillas germinadas. Los explantes se inocularon en medio de cultivo Murashige y Skoog con los diferentes reguladores del crecimiento en concentraciones predefinidas. El ADN se extrajo mediante el método CTAB a partir de plantas completas obtenidas por medio de embriogénesis somática inducida por ácido naftalenoacético (ANA) u organogénesis indirecta inducida por bencilaminopurina (BAP) o tidiazurón (TDZ). Para el RAPD, 117 muestras de ADN se amplificaron mediante diez cebadores y se seleccionaron 49 bandas puntuales entre los productos, para el estudio comparativo. Se obtuvieron un total de 5733 fragmentos, con una tasa de 5.37% de polimorfismo. ANA no generó polimorfismo y BAP fue responsable de la tasa más alta obtenida (14.28%). Se identificaron dos cebadores RAPD como marcadores para monitorear la estabilidad genética de la berenjena. El patrón polimórfico se observó solo en los clones originados en la organogénesis indirecta. Estos resultados indican la utilidad de un protocolo de monitoreo para estudios que usan berenjena clonada in vitro. Palabras clave: Solanum melongena, cultivo in vitro, variación somaclonal, RAPD, polimorfismo de ADN, fitomejoramient

Morfogênese in vitro de variedades brasileiras de cana-de-açúcar

The objective of this work was to establish in vitro systems for sugarcane plant multiplication and for regeneration from shoot apices excised from these plants. Three methods were analyzed: culture on semi-solid medium and liquid culture with or without agitation. The highest regeneration rates were obtained from cultures in liquid-medium with agitation. Shoot tips derived from these plants presented regeneration rates suitable for utilization in transformation protocols. PPT-resistant and GUS positive calluses were obtained from explants of in vitro plants of Chunnee variety inoculated with Agrobacterium tumefaciens C58C1 (pMP90) (pDUBarA9). The established in vitro culture system can be applied for transgenic sugarcane production, aiming at gene regulation studies and introduction of agronomical traits.O objetivo deste trabalho foi estabelecer sistemas de multiplicação de plantas de cana-de-açúcar in vitro e avaliar sua utilização, como material inicial, para a indução de regeneração a partir de ápices caulinares. Três métodos de cultivo foram avaliados: cultura em meio semi-sólido, cultura líquida estacionária e cultura líquida sob agitação. A taxa de multiplicação mais elevada foi alcançada por meio da cultura líquida sob agitação. Ápices caulinares, excisados dessas plantas, apresentaram taxas de regeneração in vitro compatíveis com sua utilização em protocolos de transformação. Calos resistentes a PPT e GUS-positivos foram obtidos de explantes da variedade Chunnee com inoculação de Agrobacterium tumefaciens C58C1 (pMP90) (pDUBarA9). O protocolo estabelecido a partir de cultivo in vitro pode ser utilizado para a produção de plantas transgênicas de cana-de-açúcar, visando à realização de estudos de regulação da expressão gênica, assim como à introdução de características de interesse agronômico

Evaluation of cryopreservation of Petiveria alliacea somatic embryos based on stress caused for the method used / Avaliação da criopreservação de embriões somáticos de Petiveria alliacea com base no estresse causado pelo método usado

Petiveria alliacea is a medicinal species with great potential for pharmacological use against several pathologies, including neoplasms. Many studies have been developed to optimize efficient production methodologies and long-term conservation of this species' genetic resources, with a view to phytochemical and pharmacological research. This study demonstrates the efficiency of the D-cryoplate technique, applied to somatic embryos from plants maintained in vitro. Leaf explants were inoculated in culture medium containing 20 ?M PIC and incubated under standard conditions in plant tissue culture. After 60 days, the somatic embryos induced directly on the leaf tissue surface were transferred to the multiplication medium (MS0). For cryopreservation, samples of these embryos were precultured for 24 hours in medium supplemented with sucrose (0.5M), then groups of 3-5 somatic embryos were encapsulated in calcium chloride directly in aluminum cryoplates. The cryoplates with somatic embryos adsorbed were immersed in a conditioning solution (loading) for 20 min. After being removed from loading, the somatic embryos adhered to the cryoplates were exposed to laminar flow air for different periods of time (0 to 140 min) to assess the level of dehydration. Then, samples submitted at each time were immersed in liquid nitrogen for 2 min. After this time, the cryoplates were removed and kept at room temperature for 20 min, and the somatic embryos were cultivated in MS0 medium. Evaluation after each treatment showed a high survival rate (93%) in cryopreserved somatic embryos. After 90 days of culture it was observed that somatic embryos dehydrated for 120 min showed the highest multiplication rate (32 embryos/inoculated embryo) obtained so far with these explants. The D-cryoplate technique brought innovation to established protocols representing the best option for in vitro conservation of these structures biotechnologically produced that are so promising for phytochemical and pharmacological research

Influência de substratos e de pré-tratamentos in vitro na aclimatização ex vitro de Arachis retusa

The objective of this work was to evaluate the influence of substrate and preconditioning treatments on the acclimatization of in vitro plants of Arachis retusa. Plants were transferred to Plantmax or sand, and fertilized with Hoagland's nutrient solution. Plants maintained in sand, with or without fertilizer, showed the highest survival rates. In order to evaluate the influence of in vitro preconditioning treatments, stem segments were cultured on MS medium supplemented with different sucrose concentrations. The highest survival and developmental rates were observed in plants from two accessions cultured on MS supplemented with 1.5% and 3% sucrose. Flowering and fruit production were observed after five months.O objetivo deste trabalho foi avaliar a influência de diferentes substratos e pré-tratamentos in vitro, na aclimatização de plantas in vitro de Arachis retusa. As plantas foram transferidas para Plantmax ou areia e adubadas com solução de Hoagland. Plantas mantidas em areia, com adubação ou sem adubação, apresentaram maiores taxas de sobrevivência. Para avaliação da influência de pré-tratamentos in vitro, segmentos de caule foram cultivados em meio MS suplementado com diferentes concentrações de sacarose. As maiores taxas de sobrevivência e desenvolvimento foram observadas em plantas cultivadas em sacarose a 1,5% e 3%. Depois de cinco meses, foram observadas a floração e a produção de frutos

Rescue of a non-viable accession and RAPD analysis of recovered plants of Arachis retusa

A regeneração in vitro de Arachis retusa foi avaliada visando à renovação e conservação de germoplasma. A estabilidade genética de plantas derivadas de eixos embrionários e segmentos apicais foi avaliada por RAPD. Foram analisados dez oligonucleotídeos decâmeros arbitrários, dos quais cinco foram selecionados. Noventa regiões genômicas foram avaliadas, com uma média de 18 loci por clone. Todos os segmentos amplificados foram monomórficos. Estes resultados indicam que as plantas são geneticamente estáveis nas regiões genômicas examinadas e que ambos os processos são adequados para a conservação in vitro do germoplasma de Arachis.In vitro regeneration of Arachis retusa was examined for the purpose of germplasm renewal and conservation. Random amplified polymorphic DNA (RAPD) fingerprinting was used to evaluate the genetic stability of plants derived from embryo axes and apical segments. Ten arbitrary decamer primers were screened and five of them were selected. Ninety genomic regions were evaluated, with an average of 18 loci per clone. All amplified segments were monomorphic. The results indicate that recovered plants are genetically stable at the assessed genomic regions and that both regeneration processes are suitable for in vitro germplasm preservation of Arachis species