Análise do gene OsGLR3.1 na tolerância ao déficit hídrico em tabaco

Abstract

The genetic transformation allows the insertion of genes of interest in the plant genome, making it possible, the functional characterization of the inserted gene until the acquisition of genetically modified plants more tolerant to drought. From the genetic transformation of Nicotiana tabacum, this study aimed to analyze the expression of OsGLR3.1 gene, which was identified as differentially expressed in Oryza sativa when submitted to drought. GLR is a family of glutamate receptor genes in plants are so-called because of the similarity in amino acid sequence encoding by the secondary structure and to ionotropic glutamate receptors animals (iGluRs). Tobacco leaf explants underwent the genetic transformation process via Agrobacterium tumefaciens. After cocultivation, the leaf explants were cultured on MS medium + Timentin and PPT selection agent. The coming of the explant shoots and selected by PPT were subcultured in vitro until the formation of seedlings. Confirmation of transformed plants was performed by standard PCR amplified amplicon for 413pb. The plants confirmed with the insertion of the transgene, two events had a single copy in the genome of the plant by quantitative PCR analysis. These plants and control plants, which have not undergone the transformation process were subjected to water stress experiment induced by PEG6000 (polyethylene glycol). Order to verify the expression of the transgene, the leaves was collected at time 0h, 6h and 24h for RNA extraction. After extraction and cDNA synthesis was performed quantitative PCR to assess the transgene expression profile and other genes (NtDREB2A, NtPYR1 and NtGLR3.4) determined by the formula Pfaffl. The transgene expression profile proved to be constant in transformed plants under normal condition and induced stress condition and the insertion of the gene OsGLR3.1 moderately affected the expression of other genes evaluated and became inconclusive this gene in response tolerance deficit water.