The Organic Matrix Associated with Calcium Oxalate Formation in Vitis labrusca

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9317366 Webbab Calcium oxalate formation is the predominant form of biomineralization in plants. Crystals of calcium oxalate form in higher plants in species-specific anatomical and developmental patterns, indicating that their formation is biologically controlled. Crystals generally form within the vacuole in association with an organic matrix, which accumulates prior to crystal formation and provides cellular control over the crystallization process. The long term goals of the proposed research are to understand what induces particular cells to form crystals, and how the organic matrix controls the crystal development. Because expression of the organic matrix is a key event in crystal initiation and differentiation, the proposed studios focus on the organic matrix associated with crystal formation in Vitis labrusca leaves. In particular, research is aimed in characterizing proteins in membrane enclosures which surround developing crystals. These membrane enclosures form prior to crystal initiation, and are in direct contact with the crystal s during development. In the proposed research polyclonal antibodies will be used that were developed against membrane envelope proteins in rabbits. The specific objectives of the research are: 1) to identify and characterize immunoreactive proteins in the membrane enclosures, and 2) to determine the specificity of antibodies for these proteins and for the membrane envelopes. Antibodies produced against the membrane proteins are used to isolate the individual protein components. The isolated proteins will be subjected to amino-terminal sequencing and their sequence compared with other protein sequences already present in data banks. The specificity of the proteins will be examined by immunoblotting crude protein extracts and solubilized membrane envelope proteins, as well as by immunoelectonmicroscopy. %%% Biomineralization is an important process both in plants and animals. Presently little is known of ho w crystals are initiated in plants, and about their growth. The proposed research deals primarily with fundamental problems of cell differentiation and calcium regulation in plant growth and development. It has also broad relevance outside this area as wll, and will provide information about the mechanisms of biomineralization in plants and other organisms. ***