Abstract
A mutant screen was developed to isolateArabidopsis thaliana mutants affected in the signaling pathway of phytochrome A. Mutagenized transgenic seedlings were screened for deregulated expression of a chalcone synthase promoter fused to luciferase after etiolated tissue was treated with far-red light. The efficiencies of two types of cameras used for imaging the mutant population and for isolating individual mutants were compared. The cooled CCD camera performed more efficiently than the intensified CCD camera, and when combined to a powerful analysis software, it allowed rapid and accurate selection of putative mutants. A description of image-analysis automation is presented together with theoretical and experimental comparisons of the two types of cameras.
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Abbreviations
- CAB:
-
chlorophyll a/b-binding protein
- CAB2 :
-
Lhcb1 gene inA. thaliana
- CCD:
-
charged couple device
- CHS :
-
gene encoding chalcone synthase
- Hz:
-
Hertz
- Luc :
-
gene encoding firefly luciferase
- M:
-
first generation of mutagenized seeds
- M2 :
-
generation of plants obtained by selffertilization of M
- MS:
-
plant growth medium of Murashige & Skoog (1962)
- PHYA:
-
phytochrome A
References
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Michelet, B., Chua, NH. Improvement of arabidopsis mutant screens based on luciferase imagingin planta . Plant Mol Biol Rep 14, 320–329 (1996). https://doi.org/10.1007/BF02673364
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DOI: https://doi.org/10.1007/BF02673364