• Define the role of transcription factor binding and miRNA action in regulating the root hair response to rhizobial infection.
  • Utilize proteomic approaches to examine the role of protein-protein complex formation and protein phosphorylation in the root hair response to rhizobial infection.
  • Develop and apply computational systems biology methods to study genes, proteins, non-coding RNAs, metabolites, and pathways involved in root hair rhizobial infection by integrating dissimilar experimental data.
  • Provide education and outreach opportunities to high school teachers, as well as K-12 students and college freshmen.
I.

Define the role of transcription factor binding and miRNA action in regulating the root hair response to rhizobial infection. This objective will focus on regulatory networks involved in the regulation of soybean gene expression.

We will continue to use our standardized treatment regime and root hair isolation method (see protocols). We will seek to dissect the root hair cellular responses into specific steps in the infection process.

II.
Utilize proteomic approaches to examine the role of protein-protein complex formation and protein phosphorylation in the root hair response to rhizobial infection. Eukaryotic transcription factors often mediate their effects as part of a larger protein complex. Therefore, we will utilize proteomic approaches to define the protein complexes involved in nodulation related gene expression in the root hair. Starting at the initial recognition of the Nod factor by membrane associated receptor kinases, it is clear that protein phosphorylation cascades play a critical role in the regulation of rhizobial root hair infection and subsequent nodule formation. Hence, protein phosphorylation is a key, nodal step in the nodulation regulatory circuit. Our experiments will define the complexity of these regulatory pathways.
III.

Develop and apply computational systems biology methods to study genes, proteins, non-coding RNAs, metabolites, and pathways involved in root hair rhizobial infection by integrating dissimilar experimental data. To complement the studies described in Obj. 1 and 2, we will develop and apply computational systems biology methods to study genes, proteins, non-coding RNAs, metabolites, and pathways involved in root hair rhizobial infection by integration of available data and that gathered during the course of the project.  We believe we are in a unique position in having such a wealth of data, obtained under controlled, reproducible conditions and using a single, differentiated plant cell type. However, we recognize the challenge of data integration, as there is no existing framework that can integrate such a broad spectrum of data to build a systematic model. As a result, we will adopt, improve, and develop various bioinformatics tools to construct and refine models for understanding complex regulatory mechanisms at various levels by an iterative modeling-hypothesis-experiment approach.

IV.

Provide education and outreach opportunities through research experiences and curriculum. The outreach efforts of this proposal will impact both K-12 teachers and students, as well as college freshman, through programs designed to integrate plant genome research into the curriculum. To provide long-term sustainability of these programs, the scientists involved in the research will work with the MU Office of Science Outreach (http://education.missouri.edu/orgs/oso/)\, Office of Undergraduate Research (http://undergradresearch.missouri.edu/) and other MU plant science colleagues to create four new programs that build on existing successful partnerships. The goals of the outreach programs are:

  1. Immerse high school students in research through an 8-week Research Experience program.
  2. Provide an annual one-day teacher professional development workshop for middle and high school teachers in partnership with the Missouri Partnership for Educational Renewal (MPER).
  3. Develop and implement a plant genome research module into the high school biology curriculum in collaboration with the Partnership for Research and Education of Plants (PREP) program.
  4. Establish the Freshman Research in Plant Science (FRIPS) program, a new initiative to recruit freshman students to plant science research
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