The type-IV secretion system (T4SS) is a machinery able to transfer DNA and proteins between bacteria and in certain cases also to eukaryotic cells. This thesis compared the T4SSs present in... Show moreThe type-IV secretion system (T4SS) is a machinery able to transfer DNA and proteins between bacteria and in certain cases also to eukaryotic cells. This thesis compared the T4SSs present in Agrobacterium tumefaciens and the conjugative plasmid RP4. A. tumefaciens is able to transfer DNA into plant cells and transform them genetically. We discovered that a novel hairpin structure on the terminus of the transferred DNA allows for earlier expression of its genes in the recipient plant cell. Further, we could describe the translocation signal of the relaxase protein TraI of RP4 responsible for DNA processing and transfer via the T4SS. Additionally, it was shown that the T4SS of RP4 is able to translocate proteins from one cell to another without parallel DNA transfer. Furthermore, we were able to create a hybrid relaxase created consisting of parts from A. tumefaciens (relaxase part) and RP4 (translocation signal) that was shown to be translocated via the T4SS of RP4. Show less
The type-IV secretion system (T4SS) is a machinery able to transfer DNA and proteins between bacteria and in certain cases also to eukaryotic cells. This thesis compared the T4SSs present in... Show moreThe type-IV secretion system (T4SS) is a machinery able to transfer DNA and proteins between bacteria and in certain cases also to eukaryotic cells. This thesis compared the T4SSs present in Agrobacterium tumefaciens and the conjugative plasmid RP4. A. tumefaciens is able to transfer DNA into plant cells and transform them genetically. We discovered that a novel hairpin structure on the terminus of the transferred DNA allows for earlier expression of its genes in the recipient plant cell. Further, we could describe the translocation signal of the relaxase protein TraI of RP4 responsible for DNA processing and transfer via the T4SS. Additionally, it was shown that the T4SS of RP4 is able to translocate proteins from one cell to another without parallel DNA transfer. Furthermore, we were able to create a hybrid relaxase created consisting of parts from A. tumefaciens (relaxase part) and RP4 (translocation signal) that was shown to be translocated via the T4SS of RP4. Show less
The RNA guided endonuclease based on the CRISPR/Cas system of Streptococcus pyogenus is a potent new tool for genome engineering in plants. Delivery of this system is usually done by... Show moreThe RNA guided endonuclease based on the CRISPR/Cas system of Streptococcus pyogenus is a potent new tool for genome engineering in plants. Delivery of this system is usually done by transforming plants (and other organisms) with Agrobacterium tumefaciens which transfers a T-DNA encoding the required genes through its Type IV Secretion System (T4SS). Instead, here we report the delivery of the Cas9 protein of the CRISPR/Cas system of Streptococcus pyogenus through the T4SS of Agrobacterium to Nicotiana benthamiana and Saccharomyces cerevisiae. Show less
The development of methods for the genetic modification of plants a few decades ago has provided a tremendous boost for molecular plant science. Crop plants have been generated that are... Show moreThe development of methods for the genetic modification of plants a few decades ago has provided a tremendous boost for molecular plant science. Crop plants have been generated that are resistant to insects or herbicides, or that produce useful sugars or healthy nutrients. Although the ban on growing GM crops in Europe has considerably limited the application of GM technologies, they have still contributed considerably to fundamental plant science. Especially by using the natural and very efficient mechanism of DNA transfer by the soil born bacterium Agrobacterium tumefaciens, many collections of mutant lines of model plant species such as Arabidopsis and rice have been generated, in which genes are disrupted or overexpressed by the insertion of an Agrobacterium transfer DNA (T-DNA) construct. These collections have been used in forward or reverse genetics studies to unravel the function of a gene or a family of genes in plant defense or development, and to identify the key regulators in these processes. The study described in this thesis focused on the use of one of these key regulators, the Arabidopsis AT-HOOK MOTIF NUCLEAR LOCALIZED PROTEIN 15/REJUVENATOR (AHL15/RJV), to alter developmental processes such as flowering, senescence and regeneration. Show less
