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Inducible, Cell Type-Specific Expression in Arabidopsis thaliana Through LhGR-Mediated Trans-Activation
Published on: April 19, 2019
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Efficient Plastid Transformation in Arabidopsis.
Qiguo Yu1, Kerry Ann Lutz2, Pal Maliga3,4
1Waksman Institute of Microbiology, Rutgers, the State University of New Jersey, Piscataway, New Jersey 08854.
Plant Physiology
|July 26, 2017
Summary
Plastid transformation in Arabidopsis is now 100-fold more efficient using ACC2-defective plants. This breakthrough overcomes a major hurdle, enabling advanced plastid biology research and applications in recalcitrant crops.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Genetics
Background:
- Plastid transformation is a crucial tool in plant biology but is significantly less efficient in Arabidopsis thaliana compared to Nicotiana tabacum.
- The enzyme acetyl-coenzyme A carboxylase (ACC), particularly the plastid-targeted ACC2 isoform, plays a role in fatty acid biosynthesis and influences plastid gene expression.
- Previous research indicated that mutations in ACC2 lead to hypersensitivity to spectinomycin, a common selectable marker in plastid transformation.
Purpose of the Study:
- To investigate whether disrupting ACC2 function enhances plastid transformation efficiency in Arabidopsis.
- To develop a more efficient method for plastid transformation in Arabidopsis, facilitating its use in research and crop improvement.
- To explore the potential of using ACC2 null mutants as a model for improving plastid transformation in other recalcitrant plant species.
Main Methods:
- Arabidopsis thaliana plants with null mutations in ACC2 were generated.
- These ACC2-defective plants were subjected to plastid transformation using biolistic bombardment with a vector containing the spectinomycin resistance gene (aadA) and the green fluorescence protein (GFP) gene.
- Spectinomycin resistance and GFP expression were used to identify and confirm successful plastid transformants.
Main Results:
- Plastid transformation efficiency was enhanced approximately 100-fold in ACC2-defective Arabidopsis lines (Slavice and Columbia accessions).
- Spectinomycin-resistant colonies exhibiting GFP fluorescence were successfully generated, confirming successful plastid transformation.
- The study achieved one to two plastid transformation events per bombarded sample in the ACC2 knockout backgrounds.
Conclusions:
- Disruption of ACC2 significantly enhances plastid transformation frequency in Arabidopsis, overcoming a major limitation in the field.
- The ACC2 null background provides a rational basis for developing practical plastid transformation systems in regenerable Arabidopsis accessions.
- This finding offers a template for improving plastid transformation in other crop species that are currently recalcitrant to the process.
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