The cereals, including rice, maize, wheat, barley, sorghum, and the millets, are the most important group of plants in agriculture, and environmental stress is the major limitation to their productivity. Increasing their tolerance to abiotic and biotic stress is an important goal in agriculture and considerable progress has been made in discovering the mechanisms of cell signaling that regulate the response to these stresses in plants. This project will dramatically enhance this progress by generating a protein-protein interaction database for 275 rice protein kinases, that represent each of the protein kinase subfamilies present in the recently sequenced rice genome. The protein interaction data will be obtained using both the yeast two hybrid method and a method utilizing mass spectrometry analysis of affinity-tagged protein complexes that will be purified from transgenic plants. Additionally, 100 representative orotein kinases will be mutated by either insertional mutations or RNAi silencing, and their mutant phenotypes analyzed. This combined information will available at the PlantsP website and will advance the understanding of cell signaling pathways. The knowledge of the mechanisms that plant cells use to regulate their tolerance to environmental stress will be critical for improving the agricultural productivity of the cereals. Deliveables: Data. The project data will be deposited at the PlantsP website (http://plantsp.sdsc.edu) along with the gene annotations. Full-length cDNA sequences will be deposited in Genbank and PlantsP as soon as they are completed and checked. The protein interaction data, and knockout phenotype data will be entered into the PlantsP database at no longer than six month intervals after completing the individual experiments.
Materials. The rice cDNA libraries, BD-kinase constructs, and yeast 2-hybrid bait arrays will be made available on request from the Song laboratory (Wen-Yuan Song, University of Florida). They will be advertised in a publication, which will occur in year. Availability of the libraries will also be advertised on the PlantsP website and at the annual plant phosphorylation meeting, which reaches most of the plant protein kinase community.
Transgenic rice seeds will be made available from Pam Ronald's laboratory (UC Davis) or Jian-Kang Zhu's laboratory (Univeristy of Arizona) until a suitable rice resource center is available.
