phytonematodes; Meloidogyne; RNA interference; gene silencing; Nicotiana tabacum / Isolamento de genes e construção de vetores para o uso no silenciamento gênico de Meloidogyne incognita

AUTOR(ES)

Isabela Tristan Lourenço

DATA DE PUBLICAÇÃO

2008

RESUMO

Plant parasitic nematodes are responsible for huge economic losses in world agriculture that reaches US$ 125 billion yearly. The most important of these phytopatogens, Meloidogyne incognita, has six developmental stages during the life cycle (egg, four juveniles and female). The juvenile 2 enters the host root via mechanical force and enzymatic degradation to establish the host-pathogen interaction and differentiate in apomitic adult female, which deposits 2000 eggs. Current agronomic practices have usually been unsuccessful and expensive, so the cultivation of resistant varieties is actually the most efficient way to control nematodes. In this work, it was chosen to transform icotiana tabacum using plasmidial constructions for double strand RNA expression, aiming silencing target genes Isocitrate Lyase, Arginine Kinase, 14-3-3 Protein and Heat Shock Protein 90 of Meloidogyne incognita. Four regions of target genes were selected, isolated, subcloned in RNAi vector and are being used for plant transformation. In addition, it was done an expression analysis of the four target genes, normalized with housekeeping genes β-actin e 18S rRNA, using quantitative real time PCR. This experiment showed that these genes are differentially expressed when compared during egg, J2 and female phases. The major expression difference observed was the Isocitrate Lyase gene, which is a hundred times more expressed in egg or female when compared with J2.

ASSUNTO(S)

nicotiana tabacum rna interferente fitonematóide meloidogyne silenciamento gênico biofisica molecular

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