Limits...
T-DNA insertion, plasmid rescue and integration analysis in the model mycorrhizal fungus Laccaria bicolor.

Kemppainen M, Duplessis S, Martin F, Pardo AG - Microb Biotechnol (2008)

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.

View Article: PubMed Central - PubMed

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

Related in: MedlinePlus

Laccaria bicolor transgenic strain 24 and wild type grown on P5 medium supplemented with 4 mM KNO3 as the sole N source.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3815887&req=5

f6: Laccaria bicolor transgenic strain 24 and wild type grown on P5 medium supplemented with 4 mM KNO3 as the sole N source.

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.


T-DNA insertion, plasmid rescue and integration analysis in the model mycorrhizal fungus Laccaria bicolor.

Kemppainen M, Duplessis S, Martin F, Pardo AG - Microb Biotechnol (2008)

Laccaria bicolor transgenic strain 24 and wild type grown on P5 medium supplemented with 4 mM KNO3 as the sole N source.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815887&req=5

f6: Laccaria bicolor transgenic strain 24 and wild type grown on P5 medium supplemented with 4 mM KNO3 as the sole N source.
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.

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.

View Article: PubMed Central - PubMed

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
Related in: MedlinePlus