Source: UNIVERSITY OF CALIFORNIA, DAVIS submitted to NRP
THE BIOLOGICAL ROLE OF BACTERIAL MEMBRANE VESICLES IN ADHESION AND CELL-TO-CELL SIGNALING FOR VASCULAR PLANT PATHOGENS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1022832
Grant No.
2018-67012-31497
Cumulative Award Amt.
$96,983.22
Proposal No.
2020-02528
Multistate No.
(N/A)
Project Start Date
Jun 1, 2020
Project End Date
May 31, 2021
Grant Year
2020
Program Code
[A7201]- AFRI Post Doctoral Fellowships
Recipient Organization
UNIVERSITY OF CALIFORNIA, DAVIS
410 MRAK HALL
DAVIS,CA 95616-8671
Performing Department
Plant Pathology
Non Technical Summary
Wilt-disease causing bacterial pathogens like Ralstonia colonize the water-transporting xylem vessels. Although its known that these bacteria disrupt the water-transporting function of the xylem vessels, the mechanism for this disruptiong is untested. Here we determine whether the bacteria directly clog the xylem, trigger plant host auto-immunity to clog the xylem, or damage vessels and allow air to displace the sap. To do this, we are adapting the medical imaging technology x-ray microCT to created 3D x-ray images of tomato stem from healthy and wilting tomato plants. This will allow us to definitively identify the mechanism of vascular dysfunction.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2121460110050%
2121460102050%
Goals / Objectives
Using a tomato and Ralstonia pseudosolanacearum GMI1000 model, we will characterize and visualize the spatial dynamics of xylem dysfunction. We will quantify the progressive spatial decline of xylem functionality using Licor 6400XT gas analyzer measurements of we stomatal conductance as a proxy. We will focus on several categories of leaflets for the spatial analysis: wilted leaflets, turgid leaflets co-located on a petiole with wilted leaflets, and turgid leaflets on petioles without wilt.We will follow this analysis by in depth 3D X-ray visualization of stem. We will use X-ray microcomputed tomography (X-ray μCT) to differentiate between mechanisms of xylem dysfunction: blockage by bacteria, blockage by parenchyma cell tyloses, or sap displacement by gas embolisms.
Project Methods
Standard inoculation methods will be used to directly introduce pathogenic bacteria, Ralstonia solanacearum, directly into the xylem of plants. Subsequently, LICOR measurements will be performed according to standard operating procedures. The X-ray microCT data will be reconstructed with an open source tomopy832.py script using appropriate parameters.Efforts: through laboratory instruction, postdoctoral and graduate student trainees will develop scientific proficiency in these methods.

Progress 06/01/20 to 05/31/21

Outputs
Target Audience:Target audience: plant pathologists, plant biologists, and tomato researchers. We reached them by two presentations: Lowe-Power TM. 2020. "Why do plants wilt? What X-ray computed microtomography can tell us about a model bacterial pathogen" Oct 28. Microbiology in Progress Seminar Series, UC Davis, CA. (Zoom) Lowe-Power TM. 2020. "Ralstonia and Tomato Physiology during Bacterial Wilt Disease." Oct 26. Department of Plant Pathology Seminar Series. Athens, UGA. (Zoom) Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project provided the PD with the opportunity to mentor a postdoctoral researcher, which strengthened the PD's mentoring abilities. How have the results been disseminated to communities of interest?Thus far, via two presentations, including a recorded presentation available on youtube. https://www.youtube.com/watch?v=TpFLAiaZy94&feature=youtu.be&ab_channel=Lowe-PowerLab A manuscript has been posted to a pre-print server and is currently being revised for resubmission to a journal. https://www.biorxiv.org/content/10.1101/2021.03.19.436187v1 What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Both sets of data were collected as described. The data has been analyzed and a manuscript is in preparation to report these findings to the scientific community.

Publications

  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Lowe-Power TM. 2020. "Ralstonia and Tomato Physiology during Bacterial Wilt Disease." Oct 26. Department of Plant Pathology Seminar Series. Athens, UGA. (Zoom)
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Lowe-Power TM. 2020. "Why do plants wilt? What X-ray computed microtomography can tell us about a model bacterial pathogen" Oct 28. Microbiology in Progress Seminar Series, UC Davis, CA. (Zoom)