Biotecnologia
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Programa de Pós-Graduação em Biotecnologia
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URL do programa: http://www.biotecnologia.ufes.br/pt-br/pos-graduacao/PPGBIOTEC
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- ItemEnvolvimento de microRNAs na interação Carica papaya L. e Papaya meleira vírus com potencial biotecnológico(Universidade Federal do Espírito Santo, 2015-02-26) Abreu, Paolla Mendes do Vale de; Fernandes, Patrícia Machado Bueno; Zanettini, Maria Helena Bodanese; Silva, Maite Vaslin de Freitas; Rodrigues, Silas Pessini; Fernandes, Antônio Alberto RibeiroCarica papaya L. is one of the most cultivated and consumed fruits in tropical and subtropical regions worldwide. Brazil is among the largest producers and exporters of papaya fruit. The pre-harvest diseases of papaya plants are the main limitation for fruit production. Papaya sticky disease is caused by the Papaya meleira virus (PMeV). It is a commercially important pathology in papaya culture potentially causing the complete loss of fruit production. Despite of this, little is known about the papaya interaction and response mechanisms against PMeV and there is not a papaya variety resistant to the virus. It is known that papaya 20S proteasome subunit levels of increase during PMeV infection, suggesting that proteolysis is an important feature of the plant defense response mechanisms. To date, 10,598 plant microRNAs have been identified in the Plant miRNAs Database (name of the DB), but only two microRNAs, miR162 and miR403, are from papaya. In this study, plant microRNA sequences were used to search for putative microRNAs in the papaya genome. A total of 462 microRNAs, representing 72 microRNA families, were predicted to occur in papaya. Out of these, the expression of 11 microRNAs, whose targets are known to be involved in 20S and 26S proteasomal degradation and in other stress response pathways, was estimated using real-time PCR, comparing healthy and infected papaya leaf tissues. The expression of miRNAs involved in proteasomal degradation increased in response to very low levels of PMeV titre and decreased as the viral titre increased. In contrast, biotic stress-related miRNAs levels decreased in papaya tissues infected with low virus titre and increased at high PMeV levels. Corroborating this results, analysed target genes for this miRNAs had their expression modulated in a dependent manner. With the results, some miRNAs were identified as relevant to the biotechnological application. This study represents a comprehensive prediction of miRNAs in papaya. The data presented here might help to complement the available molecular and genomic tools for the study of papaya. The differential expression of specific miRNAs and the modulation of their target genes will be helpful for understanding the particular interaction of PMeV and papaya responsible of disease development.
- ItemIndução de resistência ao Papaya meleira virus em mamoeiros(Universidade Federal do Espírito Santo, 2011-03-04) Abreu, Paolla Mendes do Vale de; Ventura, José Aires; Fernandes, Patricia Machado Bueno; Loureiro, Marcelo Ehlers; Louro, Iúri DrumondCarica papaya L. is one of the most cultivated and consumed fruits in tropical and subtropical regions of the world. Brazil is the world's largest producer of papaya and the states of Espirito Santo and Bahia are responsible for more than 70% of the Brazilian production of this fruit. Diseases of fruit trees are the main limiting factors. Papaya meleira disease, caused by the Papaya meleira virus (PMeV), is one disease that does not have a resistant cultivar. However, the PMeV genome is a doublestranded RNA (dsRNA) and it has been shown that virus resistance in plants can be activated by dsRNA molecules. After detecting the presence of dsRNA, the cell initiates a process of degradation of RNA molecules, which may be viral, thus preventing the progress of the infection. This study aimed to induce resistance to PMeV in papaya seedlings using dsRNA molecules extracted from the viral genome. Four different treatments were evaluated qualitatively and quantitatively by means of molecular diagnosis of the virus in leaf samples by RT-PCR and qRT-PCR (real-time PCR), respectively. The papaya seedlings inoculated only with high load PMeV showed severe infection in the first days post-inoculation, while the seedlings inoculated with high viral load and additional dsRNA showed a milder viral infection, suggesting a reduction in infection by the virus that causes meleira. The combined inoculation of papaya seedlings with PMeV and viral dsRNA reduced the progress of the infection.