Maize cells consist of interconnected but discrete compartments that help to maintain cellular function and order. Identifying proteins that localize to these compartments is critical to understanding developmental and physiological processes in maize, which in turn provides guiding information for crop improvement. This project makes use of recent advances in genomics to identify proteins that localize to diverse cellular compartments. The project will generate reporter lines that express proteins tagged with a fluorescent marker. Using confocal microscopy, the lines will display visual information about when and where the tagged proteins are expressed and how specific proteins may be interacting and functioning. Genes for tagging will provide complete marker coverage of cellular compartments. Several resources will be used for gene selection, including known genes from the auxin/cytokinin hormone pathway and from the RAB-mediated vesicle trafficking pathways. Gene selections will also be guided by public gene models generated from maize genome sequencing projects and by specific requests from the maize research community. The outcomes of the project will include a set of stable tagged lines expressing fluorescent protein-derived tags for 100 proteins, which will represent full coverage of cellular compartments. The tagged gene constructs will be freely available from the Jackson lab, and seeds of transgenic lines will be available from the Maize Genetics Stock Center.
Access to project outcomes: Data on characterization of the lines at the cellular level will be compiled in a localization catalog posted on a public website at http://maize.tigr.org/cellgenomics. Cell biology workshops will help train the scientific user community in analysis of the tagged lines using confocal microscopy. The project will also serve tribal community colleges in the Rocky Mountain West by enhancing education in cell and molecular biology. The project will therefore generate resources that bridge cell biology and functional genomics and will provide training across diverse scientific and learning communities. Training and outreach activities will be reported on the Plant Genome Research Outreach Portal at www.plantgdb.org/pgrop/pgrop.php.
Intellectual Merit. Corn is a major crop species that has been domesticated to grow in a full range of environments. Currently, efforts to improve crop potential depend on identifying and studying new traits that could increase yield under new and diverse growing conditions. To link traits to specific functions and genes, this project developed tools for identifying and studying trait-specific proteins within cells of the corn plant. The short-term goal of the project was to develop methods to identify molecules of interest, mark the proteins with visual tags that could be tracked and produce new lines that could be used for future study and breeding. The project successfully generated over 100 new stable lines of corn that are expressing visual markers. These seed resources can be applied to diverse breeding programs, including marker-assisted breeding and other molecular methods for crop improvement. Additionally, researchers around the world have access to research lines that advance our understanding of the function of specific proteins that control crop traits. These lines are maintained as seed resources for all users and information is publicly available through a project website. Broader Impacts. A major outcome of the project has been to promote learning and education for groups traditionally under-represented in science disciplines. The project capitalized on new advances in genetics, genomics and molecular methods to attract Native American students to science careers through a partnership with Little Big Horn College (LBHC), a Tribal College in Montana. Genetics and Molecular Genetics workshops were held annually at LBHC, the University of Wyoming or at Cold Spring Harbor Laboratory. Outcomes include recruitment of students to complete BS degrees at UW, and a high proportion of students who remain at LBHC to complete AA degrees in science disciplines. Materials and training for students and faculty at LBHC will continue to improve the recruitment and retention of students in STEM careers and improve the diversity of the science enterprise. Outcomes. The specific project outcomes include teaching and research materials for genetics instruction at a Tribal College. Biological products include more than 100 corn lines, and their associated materials, that carry visual markers for research and breeding uses. Expression of the fluorescent tags in various tissues and developmental stages were captured by imaging and provide accessible information for users. The collection of biological materials including the tagged gene constructs and seeds of the new lines are all publicly available through the Cold Spring Harbor Laboratory, the University of Wyoming and the Maize Genetics Stock Center. Seeds can be requested and datasets of fluorescent-tagged corn genes and images are compiled in a visually accessible catalog accessible via the project’s public website at http://maize.jcvi.org/cellgenomics.
