Navegando por Assunto "Piper nigrum"

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Acesso aberto (Open Access)
Extração e análise eletroforética em gel de poliacrilamida (SDS-PAGE) de proteínas totais de folhas e raízes de Piper tuberculatum
(Instituto Nacional de Pesquisas da Amazônia, 2009) SILVA, Redinaldo dos Santos; SOUZA, Claudia Regina Batista de
The Pará State is the principal Brazilian producer of black pepper (Piper nigrum Link), however, the spice’s production has been damaged by the Fusarium disease. The Fusarium solani f. sp. piperis, causal agent of this disease which attacks the plant root system promoting the root rot, leaves fall and plant death. Piperaceae plants endemic of Amazon region and resistant to F. solani f. sp. piperis infection, such as Piper tuberculatum Jacq., which has been studied in order to understand this host-pathogen interaction. In this work were evaluated five conditions for total protein extraction aiming to select suitable buffers for extraction of total proteins from P. tuberculatum leaves and roots. The conditions used to roots and leaves protein extractions were salt buffer, sucrose buffer, glycerol buffer, urea buffer and sodium phosphate buffer. Quantitative analysis showed that sucrose, glycerol and urea buffers were more efficient for leaves and roots protein extractions. SDS-PAGE analysis showed distinct band patterns in leaves and roots protein extracts obtained with different buffers. Our results could support the selection of extraction buffers for proteomics analysis in P. tuberculatum - F. solani f sp. piperis interaction studies.
Acesso aberto (Open Access)
Influência dos metabólitos secundários de Piper divaricatum da região amazônica no controle do Fusarium solani f. sp. piperis causador da fusariose em pimenta do reino
(Universidade Federal do Pará, 2014-09-18) MEIRELES, Erisléia das Neves; RAMOS, Alessandra de Rezende; http://lattes.cnpq.br/1279694874191138; SILVA, Joyce Kelly do Rosário da; http://lattes.cnpq.br/2278686174214080
The culture of black pepper (Piper nigrum L.) is one of the main agricultural activities in Pará State and it suffers serious damages caused by fusariosis, disease restrict in Brazil. This paper evaluated the antifungal activity in vitro of essential oils of species of Piper rich in phenylpropanoid before the Fusarium solani f. sp. piperis, which causes the fusariosis. The inhibition of mycelial growth by the method of agar dissemination in concentration of 5 mg.mL-1 was considered: decrease to P. aduncum (20.3%) and P. Krukoffii (31.4%); moderated to P. callosum (55.7%) and P. marginatum (70.3%) and high to P. divaricatum (93.3%). The major components identified by CG-EM were dilapiol (92.0%), safrole (78.0%), methyleugenol (75.2%) and eugenol (7.9%), apiol (80.0%), Z-isoosmorizol (44.0%) and E-anethole (22.0%), respectively. The oil of P. divaricatum and its major compounds present CIM values of 0.75 mg.mL-1. The evaluation of the effects combined of eugenol and methyl eugenol pointed eugenol as the main responsible by the activity.
Acesso aberto (Open Access)
Isolamento, caracterização e expressão em sistema bacteriano de um gene que codifica uma proteína transportadora de lipídeos de Piper nigrum
(Universidade Federal do Pará, 2012-05-11) BRITO, Wendell Upton de; SOUZA, Cláudia Regina Batista de; http://lattes.cnpq.br/3633972375952822
Black pepper (Piper nigrum L.) is one of the most widely used spices in the world belonging to the Piperaceae family, which comprises about 1400 species distributed mainly in the American tropics and Southern Asia, where this crop originated. Black pepper was introduced in Brazil in the 17th century and has been a nationally important crop since 1933. The Pará State is the main Brazilian producer of black pepper; however, the spice‘s production has been damaged by the root rot disease caused by Fusarium solani f. sp. piperis. Previous studies have revealed the identification of some cDNA sequences differentially expressed during compatible black pepper - F. solani f. sp. piperis interaction. Among them, a partial cDNA sequence coding for a putative Lipid transfer protein (LTP), which is know to play important roles in plant defense against insects and pathogens. Therefore, the aim of this work was to isolate and characterize the full-length cDNA and genomic sequences of black pepper LTP, named PnLTP. The PnLTP full-length cDNA isolated by RACE assays showed 621 bp with 32 bp and 235 bp non-coding sequences at the 5‘ and 3‘ ends, respectively. This cDNA contains a 354-bp ORF coding for a deduced protein with 117 amino acid residues that showed high identity with LTPs from different crops. Sequence analysis revealed that PnLTP contains a putative peptide signal at amino-terminal and eight cysteine residues predicted to form four disulfide bridges, which could contribute to protein stability. Alignment between genomic and cDNA sequences revealed no introns within PnLTP coding region, that is in accordance with other LTP plant genes. Finally, the mature PnLTP was expressed in bacteria system. Additional experiments will be performed in order to evaluate the ability of recombinant PnLTP in inhibit the F. solani f. sp. piperis growth.
Acesso aberto (Open Access)
Método de identificação de genes taxonomicamente restritos em dados de RNA-seq em organismo não modelo
(Universidade Federal do Pará, 2015-09-28) OLIVEIRA, Lorena Silva de; DARNET, Sylvain Henri; http://lattes.cnpq.br/4586614214029929
Black pepper (Piper nigrum) is a biotechnologically interesting plant, research target for both metabolism exploration and improvement related to phytopathological problems, in addition to understanding the evolution of basal angiosperms, ancestral group to which it belongs. With the technological revolution, the next generation sequencing offered access to genetic heritage of non model plants enabling the opening of new biotechnological perspective. The identification of non homologous genes restricted to certain species, called taxonomically restricted genes (TRGs), is a primary biotechnological target, especially in species and groups that are divergent and ancestral. This study aims to establish a method for TRGs identification from RNA-seq data and to validate the approach a dataset for black pepper. The method consists in filtering the transcripts in several stages, so that the annotated transcripts and false positives are removed, and the remaining data without molecular information are classified as potential TRGs. The application of this approach to a black pepper transcriptome dataset (35,631 transcripts) resulted in 22,661 transcripts annotated by similarity. The transcripts that were not annotated in this first analysis were processed in the TRAPID tool, resulting in 12,895 transcripts not annotated. The evaluation of transcripts for false positive detection resulted in 245 true transcripts that were analyzed for the presence of non-coding RNA, resulting in 204 unidentified transcripts. At the end of the method application 71 non annotated transcripts remained with coding regions of protein, indicating potential TRGs. The characterization of these potential TRGs in black pepper can provide new information about the molecular mechanism of this specie and perhaps elucidate pathways for the establishment of cultivars tolerant to disease.
Acesso aberto (Open Access)
Proteína PR-4 de pimenteira-do-reino (piper nigrum l.): expressão heteróloga em sistema bacteriano e avaliação funcional
(Universidade Federal do Pará, 2015-02-06) SILVA, Diehgo Tuloza da; SOUZA, Cláudia Regina Batista de; http://lattes.cnpq.br/3633972375952822
Pathogenesis-related (PR) proteins are plant proteins that are induced in response to pathogen attack. PR proteins are grouped into 17 independent families, PR-1 to PR-17. Antifungal and enzymatic activities have been described for some of these proteins. Among them, the PR-4 family comprises class I and II of plant chitinases. cDNA clone that encode a PR-4 of the class II, designated as PnPR-4, was previously isolated, in others studies, from Piper nigrum L. (black pepper). This is an important crop for Brazil, mainly in Pará state, however its production has decreased because root rot (Fusariose) disease caused by fungus Fusarium solani f. sp. Piperis. In this study, PnPR-4 cDNA clone was used for production of the recombinant protein, named PnPR-4, by bacterial expression system. Open Read Frame (ORF) of the PnPR-4 protein, was amplified from the cDNA clone by PCR assays, then ORF was linked in the expression vector pET-29(a) and introduced, by eletroporation, into E. coli Rosetta (DE3). The production of target protein, in transformed cells, was induced by 1 mM IPTG, at 37°C for 5h. After production, enzymatic activity of recombinant PnPR-4 was evaluated by Detection of Chitinase Activity after Gel Electrophoresis Polyacrylamide. Activity was evaluated in pH: 5.0 and pH: 7.8 for showing the stability of the recombinant protein. Molecular mass of Recombinant protein, PnPR-4, was 13.5kDa, according with others PR-4 produced by heterologous expression in bacterial system. In assay of enzymatic activity, PnPR-4 presented chitinolytic activity, both in pH: 5.0 or 7.8. This is a characteristic of plant quitinases, activity in a broad range of pHs. Where, optimal activity occurs between pH: 3.0 and 5.0. For PnPR-4, the optimal pH was 5.0, however in pH: 7.8 the recombinant protein had activity, demonstrating the stability of enzyme. These results showing that bacterial system of heterologous expression is effective for production of the recombinant protein PnPR-4, that it is an enzyme with chitinolytic activity and highly stable. Thus, PnPR-4, is now, a new enzyme which can be used in plant biotechnology in the combat against plant's pathogenic fungi.