Progress 08/01/22 to 01/31/23
Outputs
Target Audience:There are two major target audiences that will ultimately benefit from the project. First, the oomycete research community (more specifically collaborators from the Oomycete Molecular Genetics Network- OMGN) will have access to newly generated data corresponding to the first proteomics analyses of an oomycete not known as a plant pathogen. Second, increase in the PD Tartar's proteomics expertise will have a direct impact of the quality of education received by the student population at NSU, which includes a significant proportion of students belonging to ethnic groups traditionally underrepresented in the US academic research structure (NSU is a minority serving institution) Changes/Problems:There were no major changes or problems. The only unforeseen difference between what was accomplished and what was proposed was the timeline during the November period:shipping of samples from Florida to Minnesota was postponed until after Thanksgiving to avoid any delays related to Holiday closures, in agreement with thesuggestions from the core facilities at UMN. However, this also created further delays associated with the Mass Spectrometry machine availability, since the UMN proteomics core facilitiyreceived many samples from many different labs immediately after the Thanksgiving weekend. Overall these delays did not impact the ability of PD Tartar to complete the project and meet the proposed objectives. What opportunities for training and professional development has the project provided?Training and professional development were at the core of the project since PD Tartar was able to acquire hands-on training and become proficient at using new technologies (proteomics). Proteomics is now feasible in PD Tartar's laboratoryand part of the lab long-term objectives is to transfer that knowledge to graduate students and other lab members (including students who belong to underrepresented ethnic groups in STEM, since PD Tartar's laboratory is at a Minority-Serving Institution) in order to provide the next generation of scientists with skills and knowledge that are relevant forthe current job market in biological sciences. How have the results been disseminated to communities of interest?As noted above, the short duration of the project was not compatible with the peer-review process. However,PD Tartar aims to generate two deliverables once a thorough analysis of the data is complete. First,the proteomics MS data will be deposited in appropriate public databases (The ProteomeXchange Consortium). Second, the data analysis will be the subject of a manuscript to be submitted for peer-reviewed publication and dissemination later in 2023. These potential products do not appear yet in this final report but will acknowledge the support from NIFA. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The main goals were met during the project period. As proposed, PD Tartar was released from his teaching responsibilities at Nova Southeastern University during the Fall 2022 semester (August-December 2022), and was able to use the time to accomplish the proposed objectives: First, PD Tartar did travel to the University of Minnesota (UMN) and attended the proteomics workshop offered by the UMN Center for Mass Spectrometry and Proteomics (CMSP). The workshop occurred in August 2022 (August 16-18) and established a strong basis for PD Tartar's training in proteomics, both in terms of protein sample preparation and bioinformatics analyses. As proposed, the hands-on, in-person workshop was also complemented by additional consumption on virtual learning materials after August 2022. Second, PD Tartar was able to use the knowledge acquired during the workshop to refine his laboratory workflow, as proposed and described in the proposal. Guided by conversations with the CMSP experts that PD Tartar met during the UMN workshop, new samples were prepared in the PD laboratory from September to October 2022. Triplicatesamples corresponding to proteincomplements expressed during oomycete-insect interactions were generated and ultimately assessed for QC in early November 2022. After re-engaging with CMPS experts, these samples were shipped to UMN after the Thanksgiving break in November 2022, and Mass Spectrometry data was acquired on December 13, 2022 (some delays were created by machine availability). Since then, PD Tartar has maintained collaborative conversations with staff at UMN CMSP and initiated the bioinformatics analyses, as proposed. Overall, data analysis identified 285 proteins, with a minor rate of contaminant (5%). Analyses demonstrated that 100 proteins could be associated with the oomycete (Lagenidium giganteum) secretome, while the remainder corresponded to insect (Galleria mellonella) proteins released in the media. Mining for potential bioactive compounds through bioinformatics analyses (consistent with PD Tartar's long-term objectives) is currently underway. Overall, the main objectives were met during the short project period (6 month). Although there is currently no product associated with the award, PD Tartar aims to generate two deliverables once a thorough analysis of the data is complete. First,the proteomics MS data will be deposited in appropriate public databases (The ProteomeXchange Consortium). Second, the data analysis will be the subject of a manuscript to be submitted for peer-reviewed publication and dissemination later in 2023. These potential products do not appear yet in this final report but will acknowledge the support from NIFA.
Publications
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