The goal of the project is to find out the total protein composition of the plant nucleus, and to see whether this composition changes in different plant organs and cell types. To reach this goal, the PIs will combine their technical expertise in the areas of high-speed fluorescence-activated flow sorting of nuclei and high throughput analytical proteomics. The experiments will use flow sorting to rapidly purify nuclei from homogenates prepared from plant tissues and organs. These nuclei will be analyzed using high throughput proteomic techniques, particularly mass spectrometry, to identify as many as possible of the individual proteins contained within the nuclei. The project will develop these procedures using two plant species, the dicot model Arabidopsis thaliana, and the monocot crop Oryza sativa. These species were chosen since their complete nuclear genome sequences are available. Further experiments will examine nuclear protein composition within specific organs (roots, shoots, leaves, flowers), and within specific cells of the root (epidermis, endodermis, vascular system, meristem, and cortex). The SGER funding will be employed to determine the technical feasibility of this experimental strategy.
