A High Through-put Screen to Identify and Clone Maize and Arabidopsis Cell-wall Biogenesis-related Genes

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Discriminate analysis of Fourier Transform Infrared Spectra (FTIR) has been developed as a specific, high throughput method to identify mutations that affect plant cell wall components and architecture. This method will be employed to demonstrate the throughput rates needed to identify, in mutagenized populations of maize and Arabidopsis, a broad range of mutants in the biogenesis and dynamic alteration of plant cell wall architecture during growth and development. Both forward and reverse genetic screens will be employed. The FTIR "forward" screen will detect specific alterations in wall structure and architecture caused by the insertion of DNA 'tags", regardless of the genetic basis of the alteration. In this respect, the screen will be powerfully selective at the end-product level. Mutations to be identified could include those causing defects in wall substrate formation, wall component secretion and targeting, wall architecture and dynamics during growth. Genes disrupted in these mutants may subsequently be identified based on the insertion position of the tags. The "reverse" genetics approach will examine the effects of inserting DNA tags into genes that are already known to be related to cell wall biosynthesis. It is anticipated that this second approach will provide a broad set of cell wall mutants that will serve to further the development of FTIR spectrum libraries. These libraries will be useful to diagnose FTIR-selected mutants of unknown genetic deficiency. A major practical goal is to generate plants with genetically defined variation in cell wall composition and architecture to permit assessment of these modifications on plant development. As FTIR mutations are confirmed, the plant biology community will be informed of them through a web site and seeds and clones will be provided to the community through established stock centers.