Agrobacterium-mediated transformation (AMT) has abundantly been used for scientific research for identifying valuable traits. Agrobacterium not only transfers DNA but also translocates various virulence proteins encoded by the vir region on the Ti plasmid, which opened up the possibility to use this Agrobacterium-mediated protein translocation (AMPT) to trigger changes in plant cells without the need for introducing DNA.
In chapter 1, the factors and bottlenecks for plant transformation and regeneration are discussed. It brings forward the challenges in applying AMT and AMPT for crop improvement, and also the potential applications of AMPT in agriculture and biotechnology are explored.
Chapter 2 describes a previously developed split-GFP system and a method for more sensitive visualization of AMPT of GFP11-labeld proteins of interest in plant cells. The results show that improved codon usage and multimerization of the GFP11 label...
Show moreAgrobacterium-mediated transformation (AMT) has abundantly been used for scientific research for identifying valuable traits. Agrobacterium not only transfers DNA but also translocates various virulence proteins encoded by the vir region on the Ti plasmid, which opened up the possibility to use this Agrobacterium-mediated protein translocation (AMPT) to trigger changes in plant cells without the need for introducing DNA.
In chapter 1, the factors and bottlenecks for plant transformation and regeneration are discussed. It brings forward the challenges in applying AMT and AMPT for crop improvement, and also the potential applications of AMPT in agriculture and biotechnology are explored.
Chapter 2 describes a previously developed split-GFP system and a method for more sensitive visualization of AMPT of GFP11-labeld proteins of interest in plant cells. The results show that improved codon usage and multimerization of the GFP11 label resulted in enhanced sensitivity.
Chapter 3 describes the development of sensitive assay based on a 96-well plate reader to measure fluorescence in Agrobacterium cultures or plant extracts after AMPT or AMT. The method allowed to enhance AMT and AMPT co-cultivation conditions which achieved higher efficiency.
Chapter 4 describes how AMPT could reduce AMT recalcitrance by introducing heterologous proteins into plant cells. It is shown that AMPT of NahG or AvrPto increased both AMT and AMPT efficiency. The AMPT of AHL15 resulted in reduced senescence response and increased shoot regeneration on tobacco leaf discs.
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