Molecular Genetic Analysis of the Arabidopsis AB12 Locus

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The long term objective of this project is to identify proteins involved in abscisic acid (ABA) action and study their function by physiological, biochemical and molecular genetic analysis. Genes required for ABA action have been identified by isolating and characterizing mutants with altered sensitivity to ABA in Arabidopsis. Dr. Finkelstein's previous studies indicated that three ABI loci (abi1,abi2 and abi3) function in overlapping pathways of ABA response and the loss of function in both pathways was lethal in pollen. She will extend these studies by cloning the relevant loci, either by chromosome walking or genomic subtraction, focussing on ABI2. The specific objectives are: 1) Investigate timing and basis of loss of viability in abi2abi3 pollen, 2)Isolate new radiation induced abi mutants, 3) Isolate overlapping yeast artificial chromosomes spanning the region between the most tightly linked restriction fragment length polymorphism markers flanking the ABI1 locus. Alternatively, if any of the radiation induced mutations are allelic to abi2, she will use genomic subtraction to clone the region deleted in the mutant, 4) Identify the subset of genomic clones showing tightest linkage to ABI2 by RFLP fine mapping these clones with a panel of recombinant families, 5) Identify the ABI2 gene within this subset by complementation of the mutant phenotype, and 6) Isolate and characterize cDNA clones corresponding to the ABI2 gene.