2006 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter? This project is 100% devoted to addressing Component 1 (Functional Utilization of Plant Genomes: Translating Plant Genomics into Crop Improvement) of NP 302, "Plant Biological and Molecular Processes." The major problem being resolved is how to sustain and improve agricultural output in the face of continued population growth. Feeding the growing human population without compromising quality of life is a worldwide problem. The best agricultural land on the planet is already being utilized, and land still awaiting cultivation has the potential to be only marginally productive, limiting further agricultural gains. Agricultural biotechnology that is focused on increasing crop yield per acre of cultivated land will become increasingly important to sustain the food supply over time. Our goal is to identify and analyze the genes that control plant architecture, in order to ultimately manipulate them to aid in the improvement of agriculturally important traits such as robustness, fruit size and yield. The genes that have been identified to date as playing important roles in plant growth control appear to act in signal transduction pathways or in transcriptional regulation. However, the other components of these pathways and regulatory networks, as well as their targets, remain largely unidentified, and the molecular processes through which they control plant form are poorly understood. Elucidation and manipulation of these pathways to improve food crops is a major new frontier for agricultural biotechnology. Developing biotechnological means of improving crop yield will benefit both domestic farmers who will be able to harvest and export more crops with less effort and expense, and also overseas populations who currently or in the future will have inadequate food supplies. 2.List by year the currently approved milestones (indicators of research progress) FY2006/FY2007 Year 1. 1. Screen EMS-mutagenized clv3-3 families for enhancer and suppressor phenotypes, recover mutants and initiate backcrosses. 2. Determine contribution of the HD-ZIP genes to the jba-1D shoot meristem phenotype. 3. Identify targets of BOP1 and BOP2 transcriptional regulation. 4. Provide clones and sequences of maize ULT1 homologs to maize researchers. 5. Annotate ULT1 gene ortholog on rice genome. FY2008 Year 2. 1. Complete backcrosses and perform complementation tests. 2. Analyze the expression patterns of the seven miR166 loci. 3. Initiate BOP1 and BOP2 expression analysis. 4. Provide clones and sequences of BOP1 and BOP2 to maize researchers. 5. Annotate BOP1 and BOP2 gene orthologs on rice genome. FY2009 Year 3. 1. Perform phenotypic analysis of enhancers and suppressors. 2. Analyze the 35S::miR166 phenotypes in comparison to jba-1D. 3. Complete BOP1 and BOP2 expression analysis and initiate BOP1 yeast two hybrid candidate assays. 4. Act as consultant for maize GDB group, Plant Ontogeny group and rice TILLING project. FY2010 Year 4. 1. Generate and analyze double mutants with known meristem regulatory factors. 2. Characterize the jba-1D suppressor phenotype, initiate mapping. 3. Complete BOP1 yeast two hybrid candidate assays, initiate large-scale Y2H screen. 4. Identify orthologs of BOP1 interacting proteins in the rice genomic and maize EST sequence databases. FY2011 Year 5. 1. Map and clone CLV3-dependent enhancer/suppressor locus. 2. Complete jba-1D suppressor mapping, clone gene. 3. Complete large-scale Y2H screen and confirm positive candidates. 4. Identify ortholog(s) of the jba-1D suppressor gene in the rice genomic and maize EST sequence databases. 4a.List the single most significant research accomplishment during FY 2006. BOP1 and BOP2 Regulatory Targets This accomplishment addresses NP 302 Component 1, problem statement 1A – Advancing From Model Plants to Crop Plants. Our accomplished goal was to identify new downstream target genes of the BOP1 and BOP2 plant developmental regulatory proteins. Previous work has revealed that these two genes act early in a molecular pathway that regulates Arabidopsis leaf formation, but the specific components of this pathway are not fully known. Research to address this question was performed by Dr. Jennifer Fletcher and co-workers at the USDA-ARS Plant Gene Expression Center in Albany, CA. We used genetic and molecular experiments to show that BOP1 and BOP2 are required to control the expression of genes that determine leaf polarity. Our results demonstrate a critical role for the BOP genes in a key develop-mental regulatory pathway that affects leaf shape. Manipulation of such a pathway in crops plants may lead to increased yields, benefiting domestic farmers as well as export markets. 4b.List other significant research accomplishment(s), if any. NONE. 4c.List significant activities that support special target populations. NONE. 4d.Progress report. None. 5.Describe the major accomplishments to date and their predicted or actual impact. This project started April 6, 2006 and replaced project 5335-21000-016-00D. Please also see the report for project 5335-21000-016-00D. BOP1 and BOP2 Regulatory Targets: This accomplishment addresses National Program NP302 Component 1, problem statement 1A – Advancing From Model Plants to Crop Plants. Our accomplished goal was to identify new downstream target genes of the BOP1 and BOP2 plant developmental regulatory proteins. Previous work has revealed that these two genes act early in a molecular pathway that regulates Arabidopsis leaf formation, but the specific components of this pathway are not fully known. Research to address this question was performed by Dr. Jennifer Fletcher and co-workers at the USDA-ARS Plant Gene Expression Center in Albany, CA. We used genetic and molecular experiments to show that BOP1 and BOP2 are required to control the expression of genes that determine leaf polarity. Our results demonstrate a critical role for the BOP genes in a key developmental regulatory pathway that affects leaf shape. Manipulation of such a pathway in crops plants may lead to increased yields, benefiting domestic farmers as well as export markets. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? This project started April 6, 2006 and replaced project 5335-21000-016-00D. Please also see the report for project 5335-21000-016-00D. The science is being transferred to the broader research community and the general public through peer-reviewed publications, presentations at national and international scientific conferences, and communications in the lay media. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). This project started April 6, 2006 and replaced project 5335-21000-016-00D. Please also see the report for project 5335-21000-016-00D. Presentation entitled “Maintenance of stem cell populations in plants” at the North Carolina Biotechnology Center, Durham, NC, May 2006. Presentation entitled “MicroRNA regulation of plant stem cell populations” at the 65th Annual Meeting of the Society for Development Biology, Ann Arbor, MI, June 2006.