PAG-I Plant Genome I Conference

Town & Country Conference Center, San Diego, CA, November, 1992.


PG-I: 50pg1

PROGRESS TOWARD CLONING THE ARABIDOPSIS DET2 GENE.

Punita Nagpal, Veronique Vitart and Joanne Chory. Plant Biology Laboratory, The Salk Institute, San Diego, CA 92186-5800.


DET2 is a genetic locus involved in light-mediated early seedling development in Arabidopsis thaliana. Mutations in the DET2 gene are recessive and pleiotropic. When grown in the dark, det2 mutants display many characteristics (in morphology and gene expression) that are light-mediated in the wild-type. However, det2 mutants do not initiate the chloroplast developmental program in the dark and thus allow separation of two downstream light-regulated responses (gene expression and morphology) from a third, the development of chloroplasts. In addition to its role in de-etiolation, DET2 also appears to be involved in later stages of vegetative growth as det2 mutants have a prolonged vegetative phase, are late-flowering and show delayed leaf senescence. In our efforts to study the molecular mechanisms of signal transduction, DET2 has been mapped to chromosome 2 and chromosome walking experiments have allowed us to localize it to a 150 kb YAC. The experiments to delineate the gene further by walking across the YAC genetically with cosmid and lambda clones are in progress. Cosmid clones are being used to transform det2/det2 mutants and test complementation and also to screen a cDNA library made from etiolated wild-type tissue. In parallel, we have used these inserts to map a genomic rearrangement that has occurred in a new promising det2 allele obtained after fast-neutron irradiation. Our most recent progress toward identifying the gene will be presented.


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