A gene coding for green fluorescent protein (GFP) that is fused to a mitochondrial targeting signal has been constructed. Expression of this gene allows direct observation of the dynamics of mitochondria in vivo in living plant tissues under fluorescence microscopy without further staining or substrate application, and enables us to screen mutants with altered mitochondrial morphology. A chimeric gene (DIPS-GFP) was constructed in which a mitochondrial targeting signal from the ATPdelta gene and a coding region of the GFP gene were fused and placed under the control of the CaMV 35S promoter. DIPS-GFP was introduced into the T-DNA region of a binary vector, and transformed into Arabidopsis plants by Agrobacterium-mediated transformation. While the insertion of DIPS-GFP into the nuclear genome allows us to observe mitochondria in vivo, a mutant plant with altered mitochondrial status can be obtained at the same time by T-DNA mutagenesis.
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