Laboratorio Of Biologia Celular E Tecidual

São José dos Campos, Brazil

Laboratorio Of Biologia Celular E Tecidual

São José dos Campos, Brazil
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Baldotto L.E.B.,Federal University of Viçosa | Baldotto M.A.,UFV Campus Florestal | Canellas L.P.,Laboratorio Of Solos | Bressan-Smith R.,Laboratorio Of Genetica E Melhoramento Of Plantas | Olivares F.L.,Laboratorio Of Biologia Celular E Tecidual
Revista Brasileira de Ciencia do Solo | Year: 2010

In vitro propagation of pineapple produces uniform and disease-free plantlets, but requires a long period of acclimatization before transplanting to the field. Quicker adaptation to the ex vitro environment and growth acceleration of pineapple plantlets are prerequisites for the production of a greater amount of vigorous, well-rooted planting material. The combination of humic acids and endophytic bacteria could be a useful technological approach to reduce the critical period of acclimatization. The aim of this study was to evaluate the initial performance of tissue-cultured pineapple variety Vitória in response to application of humic acids isolated from vermicompost and plant growth-promoting bacteria (Burkholderia spp.) during greenhouse acclimatization. The basal leaf axils were treated with humic acids while roots were immersed in bacterial medium. Humic acids and bacteria application improved shoot growth (14 and 102%, respectively), compared with the control; the effect of the combined treatment was most pronounced (147%). Likewise, humic acids increased root growth by 50%, bacteria by 81% and the combined treatment by 105%. Inoculation was found to significantly increase the accumulation of N (115%), P (112%) and K (69%) in pineapple leaves. Pineapple growth was influenced by inoculation with Burkholderia spp., and further improved in combination with humic acids, resulting in higher shoot and root biomass as well as nutrient contents (N 132%, P 131%, K 80%) than in uninoculated plantlets. The stability and increased consistency of the host plant response to bacterization in the presence of humic substances indicate a promising biotechnological tool to improve growth and adaptation of pineapple plantlets to the ex vitro environment.


Vale E.M.,State University of Norte Fluminense | Reis R.S.,State University of Norte Fluminense | Santa-Catarina C.,Laboratorio Of Biologia Celular E Tecidual | Silveira V.,State University of Norte Fluminense
Scientia Horticulturae | Year: 2016

Heterosis describes the superior performance of a heterozygous F1-hybrid in comparison with the average performance of the parental lines for a given trait in a given environment and is the result of the effects of non-additive genes. In the present study, proteins from the primary roots of the papaya (Carica papaya L.) hybrid JS12 × São Mateus and its parental inbred lines were analyzed using proteomic analyses combining the shotgun method and nanoESI-HDMSE technology. A total of 955 proteins were identified by the shotgun method, among which 261 exhibited a trend toward heterosis in the hybrid compared with the mid-parents. Non-additive proteins were divided into “above high-parent” (16.1%), “high-parent” (6.5%), “low-parent” (22.2%), and “below low-parent” (55.2%) abundance patterns. The results revealed a decrease in proteins involved in energy-consuming processes such as protein metabolism and an increase in root development proteins such as those involved in auxin polar transport and signaling regulation. The findings suggest that the hybrid possesses an optimization mechanism for protein synthesis that results in substantial improvements in cellular energy efficiency and phenotypic performance. Therefore, this study may contribute to a better understanding of the molecular basis of heterosis in papaya. © 2016 Elsevier B.V.


PubMed | Federal University of Rio de Janeiro, State University of Rio de Janeiro, Laboratorio Of Biologia Celular E Tecidual, State University of Norte Fluminense and 2 more.
Type: Journal Article | Journal: Memorias do Instituto Oswaldo Cruz | Year: 2015

Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain) or orally infected with cysts (ME-49 strain). Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis.


PubMed | Federal University of Rio de Janeiro, State University of Rio de Janeiro, Laboratorio Of Biologia Celular E Tecidual, State University of Norte Fluminense and 2 more.
Type: | Journal: Memorias do Instituto Oswaldo Cruz | Year: 2015

Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain) or orally infected with cysts (ME-49 strain). Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis.


Bagatini P.B.,Laboratorio Of Biologia Celular E Tecidual | Saur L.,Laboratorio Of Biologia Celular E Tecidual | Rodrigues M.F.,Laboratorio Of Biologia Celular E Tecidual | Bernardino G.C.,Laboratorio Of Biologia Celular E Tecidual | And 8 more authors.
Invertebrate Neuroscience | Year: 2011

Studies have suggested that neuronal loss in Parkinson's disease (PD) could be related to the pacemaker activity of the substantia nigra pars compacta generated by L-type Cav 1.3 calcium channels, which progressively substitute voltage-dependent sodium channels in this region during aging. Besides this mechanism, which leads to increases in intracellular calcium, other factors are also known to play a role in dopaminergic cell death due to overproduction of reactive oxygen species. Thus, dihydropyridines, a class of calcium channel blockers, and resveratrol, a polyphenol that presents antioxidant properties, may represent therapeutic alternatives for the prevention of PD. In the present study, we tested the effects of the dihydropyridines,isradipine, nifedipine, and nimodipineand of resveratrol upon locomotor behavior in Drosophila melanogaster. As previously described, paraquat induced parkinsonian-like motordeficits. Moreover, none of the drugs tested were able to prevent the motor deficits produced by paraquat. Additionally, isradipine, nifedipine, resveratrol, and ethanol (vehicle), when used in isolation, induced motor deficits in flies. This study is the first demonstration that dyhidropyridines and resveratrol are unable to reverse the locomotor impairments induced by paraquat in Drosophila melanogaster. © Springer-Verlag 2011.


Vale E.D.M.,State University of Norte Fluminense | Heringer A.S.,State University of Norte Fluminense | Barroso T.,State University of Norte Fluminense | Ferreira A.T.D.S.,Instituto Oswaldo Cruz Instituto Oswaldo Cruz Ioc Fiocruz | And 4 more authors.
Proteome Science | Year: 2014

Background: Somatic embryogenesis is a complex process regulated by numerous factors. The identification of proteins that are differentially expressed during plant development could result in the development of molecular markers of plant metabolism and provide information contributing to the monitoring and understanding of different biological responses. In addition, the identification of molecular markers could lead to the optimization of protocols allowing the use of biotechnology for papaya propagation and reproduction. This work aimed to investigate the effects of polyethylene glycol (PEG) on somatic embryo development and the protein expression profile during somatic embryo maturation in papaya (Carica papaya L.).Results: The maturation treatment supplemented with 6% PEG (PEG6) resulted in the greatest number of somatic embryos and induced differential protein expression compared with cultures grown under the control treatment. Among 135 spots selected for MS/MS analysis, 76 spots were successfully identified, 38 of which were common to both treatments, while 14 spots were unique to the control treatment, and 24 spots were unique to the PEG6 treatment. The identified proteins were assigned to seven categories or were unclassified. The most representative class of proteins observed in the control treatment was associated with the stress response (25.8%), while those under PEG6 treatment were carbohydrate and energy metabolism (18.4%) and the stress response (18.4%).Conclusions: The differential expression of three proteins (enolase, esterase and ADH3) induced by PEG6 treatment could play an important role in maturation, and these proteins could be characterized as candidate biomarkers of somatic embryogenesis in papaya. © 2014 Vale et al.; licensee BioMed Central Ltd.


PubMed | State University of Norte Fluminense, Waters Corporation, University of Sao Paulo and Laboratorio Of Biologia Celular E Tecidual
Type: Comparative Study | Journal: PloS one | Year: 2015

The development of somatic cells in to embryogenic cells occurs in several stages and ends in somatic embryo formation, though most of these biochemical and molecular changes have yet to be elucidated. Somatic embryogenesis coupled with genetic transformation could be a biotechnological tool to improve potential crop yields potential in sugarcane cultivars. The objective of this study was to observe somatic embryo development and to identify differentially expressed proteins in embryogenic (E) and non-embryogenic (NE) callus during maturation treatment. E and NE callus were cultured on maturation culture medium supplemented with different concentrations (0.0, 0.75, 1.5 and 2.0 g L(-1)) of activated charcoal (AC). Somatic embryo formation and differential protein expression were evaluated at days 0 and 21 using shotgun proteomic analyses. Treatment with 1.5 g L(-1) AC resulted in higher somatic embryo maturation rates (158 somatic embryos in 14 days) in E callus but has no effect in NE callus. A total of 752 co-expressed proteins were identified through the SUCEST (The Sugarcane EST Project), including many housekeeping proteins. E callus showed 65 exclusive proteins on day 0, including dehydrogenase, desiccation-related protein, callose synthase 1 and nitric oxide synthase. After 21 days on maturation treatment, 14 exclusive proteins were identified in E callus, including catalase and secreted protein. NE callus showed 23 exclusive proteins on day 0 and 10 exclusive proteins after 21 days on maturation treatment, including many proteins related to protein degradation. The induction of maturation leads to somatic embryo development, which likely depends on the expression of specific proteins throughout the process, as seen in E callus under maturation treatment. On the other hand, some exclusive proteins can also specifically prevent of somatic embryos development, as seen in the NE callus.


Baldotto L.E.B.,State University of Norte Fluminense | Baldotto M.A.,Laboratorio Of Solos | Olivares F.L.,Laboratorio Of Biologia Celular E Tecidual | Viana A.P.,Laboratorio Of Genetica E Melhoramento Of Plantas | Bressan-Smith R.,Laboratorio Of Genetica E Melhoramento Of Plantas
Revista Brasileira de Ciencia do Solo | Year: 2010

Pineapple (Ananas comosus L. Merrill) in vitro propagation results in the production of a large quantity of healthy and homogeneous plantlets. Despite these advantages, the long time required for acclimatization makes this agricultural practice too costly. Growth acceleration of plantlets by inoculation with endophytic and epiphytic diazotrophic bacteria may be useful to reduce this period. The objectives of this study were to evaluate the potential of 20 diazotrophic bacteria strains in synthesizing indole, solubilizing Ca phosphate and Zn oxide and acting against the fungus Fusarium subglutinans f. sp. ananas, and subsequently, evaluate the performance of pineapple 'Vitória' propagated by tissue culture in response to the application of these isolates during the acclimatization period in a greenhouse. Shoot and root growth characteristics and leaf nutrient content of pineapple were evaluated. Results showed differences in the ability of the bacteria strains to synthesize indole, solubilize Ca phosphate and Zn oxide and act against Fusarium. Differences in the growth-promoting capacity of the shoot and root system of bacteria and leaf accumulation of N, P, K, Ca and Mg in pineapple were also shown. Inoculation with diazotrophic bacteria can promote the growth of plantlets during the acclimatization period, improving the adaptation of pineapple to the exvitro environment.


Reis R.S.,State University of Norte Fluminense | Vale E.D.M.,State University of Norte Fluminense | Heringer A.S.,State University of Norte Fluminense | Santa-Catarina C.,Laboratorio Of Biologia Celular E Tecidual | Silveira V.,State University of Norte Fluminense
Journal of Proteomics | Year: 2016

Somatic embryogenesis, an important biotechnological technique, has great potential for application in sugarcane breeding and micropropagation. Polyamines have been associated with the regulation of several physiological processes, including the acquisition of embryogenic competence and somatic embryogenesis. In this study, we used a proteomic approach to evaluate the effects of exogenous polyamine on sugarcane somatic embryo development to better understand this process. Embryogenic cultures were treated with different concentrations of putrescine, spermidine, and spermine. Proteomic analyses combined the shotgun method and the nanoESI-HDMSE technology. Among polyamines, 500μM putrescine gave rise to the highest number of somatic embryos; however, no differences in the amount of fresh matter were observed between polyamines and control. Differences in protein abundance profiles resulting from the effect of 500μM putrescine on sugarcane somatic embryo maturation were observed. Proteomic analyses of putrescine and control treatment showed differences in the abundances of proteins related to somatic embryogenesis, such as arabinogalactan proteins, peroxidases, heat shock proteins, glutathione s-transferases, late embryogenesis abundant proteins, and 14-3-3 proteins. These results show that putrescine and the identified proteins play important roles in protecting the cells against an in vitro stress environment, contributing to the formation of somatic embryos during the maturation treatment. Biological significance: Despite all studies with somatic embryogenesis, the molecular mechanisms controlling the process have not been completely understood. In this study, we highlighted the effects of the polyamine putrescine on somatic embryogenesis of sugarcane and the differentially abundant proteins related to somatic embryo development. We identified six groups of important stress related proteins that are involved in the adaptation of cells to the stress environment of in vitro culture and may also be part of the mechanisms associated to the somatic embryogenesis process. Therefore, our research is trying to understand the complexity of how one single somatic cell becomes a whole plant. © 2015 Elsevier B.V.


PubMed | State University of Norte Fluminense and Laboratorio Of Biologia Celular E Tecidual
Type: | Journal: Journal of proteomics | Year: 2015

Somatic embryogenesis, an important biotechnological technique, has great potential for application in sugarcane breeding and micropropagation. Polyamines have been associated with the regulation of several physiological processes, including the acquisition of embryogenic competence and somatic embryogenesis. In this study, we used a proteomic approach to evaluate the effects of exogenous polyamine on sugarcane somatic embryo development to better understand this process. Embryogenic cultures were treated with different concentrations of putrescine, spermidine, and spermine. Proteomic analyses combined the shotgun method and the nanoESI-HDMS(E) technology. Among polyamines, 500 M putrescine gave rise to the highest number of somatic embryos; however, no differences in the amount of fresh matter were observed between polyamines and control. Differences in protein abundance profiles resulting from the effect of 500 M putrescine on sugarcane somatic embryo maturation were observed. Proteomic analyses of putrescine and control treatment showed differences in the abundances of proteins related to somatic embryogenesis, such as arabinogalactan proteins, peroxidases, heat shock proteins, glutathione s-transferases, late embryogenesis abundant proteins, and 14-3-3 proteins. These results show that putrescine and the identified proteins play important roles in protecting the cells against an in vitro stress environment, contributing to the formation of somatic embryos during the maturation treatment.Despite all studies with somatic embryogenesis, the molecular mechanisms controlling the process have not been completely understood. In this study, we highlighted the effects of the polyamine putrescine on somatic embryogenesis of sugarcane and the differentially abundant proteins related to somatic embryo development. We identified six groups of important stress related proteins that are involved in the adaptation of cells to the stress environment of in vitro culture and may also be part of the mechanisms associated to the somatic embryogenesis process. Therefore, our research is trying to understand the complexity of how one single somatic cell becomes a whole plant.

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