T-DNA insertion, plasmid rescue and integration analysis in the model mycorrhizal fungus Laccaria bicolor.
Bottom Line: Our results demonstrate that the plasmid rescue approach can be used for resolving T-DNA integration sites in Laccaria.Neither obvious sequence similarities were found between these sites and the T-DNA borders indicating non-homologous integration of the transgenes.Agrobacterium-mediated gene transfer is a powerful tool that can be used for functional gene studies in Laccaria and will be helpful along with plasmid rescue in searching for relevant fungal genes involved in the symbiotic process.
Affiliation: Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, (B1876BXD) Bernal, Provincia de Buenos Aires, Argentina.Show MeSH
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Mentions: Supplementary phenotyping analysis was then carried out to study closer the dikaryotic transgenic strain 24. While on P5 with ammonium this strain had not shown an altered growth, interestingly the same medium supplemented with 4 mM KNO3 as the sole N source revealed a phenotype clearly distinguishable from the wild‐type strain. On this medium the transgenic strain grew more vigorously and had at least twice more hyphae penetrating the agar than the wild type while the radial growth rate was not altered (Fig. 6). These observations finally confirm that single tagged mutagenesis of dikaryotic Laccaria indeed can generate functional mutants with detectable phenotypes when observed under variable growth conditions.
Affiliation: Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, (B1876BXD) Bernal, Provincia de Buenos Aires, Argentina.