1a.Objectives (from AD-416):
In this project, we propose to investigate the proteomes of leaf and root tissues in response to different levels and duration of drought stress using isobaric stable isotope containing mass tags. There are a number of specific objectives including:. 1)To construct protein function networks for stress-regulated proteins;. 2)To identify additional transcriptional factors and signal transduction factors that are putative partners of the identified proteins through database-mining strategies;. 3)To develop mathematical models for plant growth response to drought stress;. 4)To build an integrated model for protein expression and growth properties; 5). To set up a genetic transformation system for switchgrass to produce transgenic plants; . 6)To train a team of faculties and students through collaboration with USDA scientist; 7). To develop web-based resources (public data repositories and Twitter, Blog, etc.) to disseminate real-time information to the scientific community and the general public.
1b.Approach (from AD-416):
Specific activities performed in the ARS labs will include a) a comparative proteomic analysis using an 8-plex iTRAQ strategy of leaf and root tissues from switchgrass subjected to drought stess of different durations.
This project provides the means to continue a long standing mentoring/capacity building and research relationship between Tennessee State University (TSU, an 1890 University) and the R. W. Holley Center for Agriculture and Health (RWHC). Although this project is still in its earliest phase (funds were made available on 4/15/2013) considerable progress has been made with respect to the twin goals of training and research. From 5/23/2013-7/28/2013 a professor and 3 of her students were in residence at the RWHC. They were in daily contact with RWHC scientists and received individualized hands-on training from the expert practitioners of the RWHC staff specific to the proteomics aspects of this project. Additionally, the RWHC sponsored these individuals for a 3-day workshop on proteomics run jointly by the RWHC and Cornell University that provided more generalize training in proteomics.
Research progress was similarly impressive. Three multiplexed samples were prepared from the proteins isolated from the various biological materials (roots and leaves) prepared at TSU during the spring. These were analyzed by high performance mass spectrometry. We are currently evaluating the available genomic resources for their suitability for proteomic analysis. Although the data is preliminary, it appears quite promising with > 2000 proteins identified and quantified in each sample. The functional aspects of this project have yet to begin.