The alcohol dehydrogenase-l (Adhl) gene is one of 10 major polypeptides (ANPs) synthesized in response to anaerobic stress. The system is accessible to recombinant DNA technology much as is the heat-shock system in animals. The overriding objective is to understand the behavior and structure of every component of a "typical" gene in higher organisms and how genes are coordinated quantitatively and organ specifically during development. We will continue to induce, recover via pollen selection, and analyze several Adhl mutants altered in regulatory parameters such as timing, rate, organ-specificity, stability, or circuitry of expression. Using interesting mutants, including unstable "controlling element" alleles, we will discover which step is gene expression -- e.g., tranascription, RNA-processing, translation -- is responsible for the regulatory behavior. Among other tests, quantitative Northern hybridization techniques and genetic fine structure analyses will be used. We will also characterize the protein-level molecular biology of the 10 major ANPs and polypeptides specific to the transition from an aerobic to an anaerobic environment (TPs); enzymatic functions for the ANPs and TPs will be sought. In addition, we plan to begin a formal, quantitative genetic study treating the anaerobic genes as a co-adapted complex, and to continue development of a protoplast transformation system using Adhl as the selectable marker.