Source: UNIVERSITY OF MISSOURI submitted to
UNDERSTANDING PLANT ROOT BORDER CELLS AND THEIR SECRETIONS IN MEDIATING RHIZOSPHERE MICROBIOME DYNAMICS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
NEW
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1030651
Grant No.
2023-67012-40304
Project No.
MO00080987
Proposal No.
2022-09748
Multistate No.
(N/A)
Program Code
A1402
Project Start Date
Jul 1, 2023
Project End Date
Jun 30, 2025
Grant Year
2023
Project Director
Kranawetter, C.
Recipient Organization
UNIVERSITY OF MISSOURI
(N/A)
COLUMBIA,MO 65211
Performing Department
(N/A)
Non Technical Summary
We seek to understand the chemical mechanisms by which plants engage in beneficial microbial relationships and defend against pathogens, which will benefit food quality, yield, and supply. The frontier of root defense against pathogens is led by plant root border cells, which are poised at the interface between root and soil. Border cells are a specialized cell type located at the root tips of most plant species. They violate traditional knowledge of plant cells in that they are not formally attached to the plant root tip. Instead, border cells adhere to the root tip through a complex assortment of secretions. Since they are not properly attached to the root, border cells can be removed upon exposure to water, and they are rapidly replaced within 24 hours after initial removal to be held in place once again by their secretions. Border cell secretions include DNA (which is highly unique as a secretion), proteins, mucilage, and metabolites. In general, metabolites encompass a wide range of compounds necessary for life and growth. Recognizable metabolites to the general public might include glucose and caffeine, but there are thousands of others. While helping to adhere border cells to the root tip, these secretions also form a sticky "trap" that captures microbes, preventing pathogens from accessing the root tip and preventing infection. Furthermore, the various proteins and known metabolites possess beneficial and/or chemically protective, defense related properties. Collectively, border cells are essential in protection of the root tip (which is the base of all root growth, fragile, and susceptible to lethal infection) from invading pathogenic microbes.We hypothesize that border cells directly manipulate the surrounding soil to create a favorable local chemical environment for root/plant health through secretion of metabolites. In this, our primary goals are to establish the identities of border cell secreted/released metabolites and their secretory pathways. While the metabolites arising from both border cells and the root tip have been studied previously, the specific secretions arising from border cells remains unknown. In our study, we aim to isolate border cells from the root tips of Medicago truncatula (a relative to alfalfa) in custom bioreactors. After isolation, we will harvest the secretions arising specifically from border cells to identify compounds involved in beneficial or defense-related functions. Furthermore, we will also study the specific mechanisms by which border cells secrete these compounds. Ultimately, our studies will generate knowledge empowering methods to create healthier plants with higher food yields.
Animal Health Component
0%
Research Effort Categories
Basic
50%
Applied
25%
Developmental
25%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
20614191000100%
Knowledge Area
206 - Basic Plant Biology;

Subject Of Investigation
1419 - Leguminous vegetables, general/other;

Field Of Science
1000 - Biochemistry and biophysics;
Goals / Objectives
Overarching GoalThe overarching goals of this project are to form the foundation of border cell secretory activities in rhizosphere manipulation and develop multidisciplinary skills for the post-doctoral fellow.Project ObjectivesTrainingTheoretical, hands-on operational learning, and applications of Trapped Ion Mobility Spectroscopy (TIMs) coupled to a hybrid quadrupole time-of-flight mass spectrometer (QToF-MS) in nontargeted metabolomics experiments.Milestones for training objective one include:Receiving training from Bruker personnel in proper operation and management of the Sumner lab TIMS-QToF instrument.Analysis of border cell secretions using UHPLC-TIMS-QToF-MS.Utilization of border cell secretion TIMS-QToF data to identify secreted metabolites and derive targets for bioactivity assays.Application of qRT-PCR to Medicago truncatula root border cells.Milestones for training objective two include:Reaching out to collaborators on campus for instructions in proper qRT-PCR usage and methods development.Conduction of experimental protocols we develop for specific application to border cells.Working with our collaborators on proper protocols for data publication.Probing for new proteins in Medicago truncatula root border cells using immunoblots.Milestones for training objective three include:Collaboration with immunoblot experts on MU campus to learn proper techniques for performing immunoblots.Designing new protocols to make border cells suitable for immunoblot analyses.Performing preliminary immunoblot analyses utilizing a common cytosolic marker as a proof-of-concept study.Collaborating with other researchers on campus to design antibodies (if necessary) or order antibodies to probe candidate proteins associated with specialized metabolite secretion.Collaborating with other laboratories to learn proper data presentation methods.Employment of Bioactive Metabolites in Bacterial Growth Inhibition Assays.Milestones for training objective four include:Performing background research on proper materials/methods for conducting growth inhibition assays (protocols for these assays are readily available).Conducting the growth inhibition assays, optimize, and observe the results.Collaboration and literature gathering to learn proper publication methods.ResearchEstablishment and identification of secreted metabolites arising specifically from isolated border cells of Medicago truncatula under non-elicted (baseline) and microbial elicted conditions using Ultra High Performance Liquid Chromatography (UHPLC) coupled to Trapped Ion Mobility Spectrometry-Time of Flight-Mass Spectrometry (TIMs-ToF-MS). Ultimately this will fill the knowledge gap of border cell specific contributions to rhizosphere chemistry.Milestones for research objective one include:Development of a protocol for collection of border cell secreted metabolites.Analysis of secreted compounds by UHPLC-TIMS-ToF-MS.Identification of newly secreted metabolites for the purpose of this objective and for later bioactivity studies in objective three.Obtainment (via commercial vendors) or in-house purification of newly secreted metabolites for the purpose of this objective and bioactivity studies in objectiveEstablishment of the mechanisms of specialized metabolite secretion by probing border cell extracts for changing ABC and MATE transporters associated with movement of specialized metabolites utilizing Quantitative Reverse Transcription- Polymerase Chain Reaction (qRT-PCR) and Immunoblot analysis. This will ultimately establish border cell mechanisms of specialized metabolite secretion.Milestones for research objective two include:Development of a protocol for collection of border cell RNA and execution of qRT-PCR experiments.Execution of qRT-PCR experiments.Development of a protocol for extracting proteins from border cells.Design or purchase of antibodies targeting specific transporters (including trial/error in antibody design).Execution ofimmunoblot experiments.Establishment of secreted specialized metabolite roles in relation to Sinorhizobium meliloti (a symbiont of Medicago truncatula) and Pseudomonas syringae pv. tomato DC3000 (a pathogen known to infect Medicago truncatula) using growth inhibition assays. This will ultimately provide insight into the roles of specific phytochemicals in plant defense and symbiosis.Milestones for research objective three include:Acquiring Sinorhizobium meliloti and Pseudomonas syringae pv. DC3000.Culturing and storage of glycerol stocks the aforementioned bacteria using well established medias and methodologies.Culturing of bacteria for growth inhibition assays.Completion of growth inhibitions assays using wide-range dilutions of a minimum of three pure specialized metabolites from research objective one.
Project Methods
General Research MethodsWe will profile and identify secreted metabolites that arise specifically from border cells of Medicago truncatula under non-elicited and microbial elicited conditions using Ultra High Performance Liquid Chromatography (UHPLC) coupled to Trapped Ion Mobility Spectrometry-Quadrupole Time of Flight-Mass Spectrometry (TIMs-QToF-MS) to understand border cell specific contributions to rhizosphere chemistry.M. truncatula seeds will be chemically scarified, sterilized, plated on 1% w/v water agar overlaid with filter paper, and grown for three days in the dark. Border cells will be harvested and loaded into a flow-through bioreactor assembly for metabolite trapping/harvest. Negative control secretions will be examined using border cells under baseline secretion conditions (i.e. no application of eliciting materials). Positive control secretions will be performed using Methyl-Jasmonate (Me-JA) (a known initiator of specialized metabolite secretion in M. truncatula). This same procedure will be performed using 80% Methanol:20% water extracts of S. meliloti or P. syringae (essentially small molecule extracts). A downstream Solid Phase Extraction (SPE) cartridge will be connected for secreted metabolite collection. Secreted metabolites will be trapped over 0, 0-8, 8-24, and 24-48 hours, providing insight into border cell temporal secretion patterns. Samples will be analyzed by Ultra High Performance Liquid Chromatography-Trapped Ion Mobility Spectroscopy-Quadrupole Time of Flight-Mass Spectrometry (UHPLC-TIMs-QToF-MS). Data will be processed utilizing Bruker DataAnalysis and Metaboscape software tools. Sample differences will be visualized using statistical analyses such as Principal Component Analysis (PCA) along with Partial Least Squares-Discriminant Analysis (PLS-DA). The top five metabolites of highest intensity variation will be identified for use in specific aim 3.We will examine mechanisms of secondary metabolite secretion by probing border cell extracts for changing ABC and MATE transporters associated with movement of secondary metabolites utilizing Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and Immunoblot analysis to determine mechanisms of metabolite secretion.M. truncatula seeds will be chemically scarified, sterilized, plated on 1% w/v water agar overlaid with filter paper, and grown for three days in the dark. For both qRT-PCR and immunoblot analyses, border cells will be harvested and allowed to rest overnight to dissipate stress responses. Border cells will be elicited using MeJA in DMSO diluted into S&H basal salts medium. Negative controls will use DMSO only. A time-based series (0, 0-8, 8-24, 24-48 hours) will be employed after which samples will be flash frozen and lyophilized. Dried tissue samples will undergo qRT-PCR according to manufacturer protocols (Power SYBR Green, Life Technologies or related), focusing on genes associated with ATP Binding Cassette and Multidrug and Toxins Efflux (MATE) transport families, which have known activities in primary and secondary metabolite transport. Genes of interest have been identified from available literature. Identified candidate genes will be linked to their overall roles in metabolite secretion by probing for their protein products utilizing immunoblots. Border cells will be analyzed under elicited and baseline conditions as described for qRT-PCR experiments, checking for protein product overexpression under elicited conditions. Antibodies will be purchased or designed targeting the protein products of earlier qRT-PCR targets.We will investigate the role of secreted metabolites in relation to Sinorhizobium meliloti and Pseudomonas syringae pv. tomato DC3000 using growth inhibition assays to understand the roles of specific phytochemicals in plant defense and symbiosis.Cultures for S. meliloti and P. syringae pv. tomato DC3000 are currently in our possession. Metabolites that were identified in specific aim one will be purchased commercially or purified in our lab. Bacteria will be streaked on medium plates after which filter paper discs soaked in a solution of a compound of interest will be placed on smeared regions. Stock solution concentrations will be varied to observe effects on each microbe and define an optimal concentration. Comparison to a positive control (using filter paper discs soaked in antibiotics appropriate for each bacterial species) and negative control (filter paper discs soaked in sterile water) will take place. Results will be documented by photographing culture plates and collecting representative samples for cell viability counts.Evaluation PlansTraining Evaluation PlanProject Advisory BoardAdvisory board meetings will take place twice per year (at a minimum). These meetings will serve as an opportunity to provide research updates and progress on professional development goals. In turn, the advisory board will provide insight into experimental goals and guidance in support of the project director's professional development.Individual Evaluation with Project MentorWeekly meetings with project mentor will ensure the project director has readily accessible guidance for experiments and professional development activities.Research Evaluation PlanAim 1:Successful development of a protocol for collection of border cell secreted metabolites.Analysis of secreted compounds by UHPLC-TIMS-ToF-MSIdentification of newly secreted metabolites for later bioactivity studies.Purification or purchase of target bioactive border cell secreted metabolites for utilization in growth inhibition assays outlined in Specific Aim 3.Success indicator: Establishment of border cell secretion of bioactive metabolites under baseline and microbial extract elicited conditions.Aim 2:Development of a protocol for collection of border cell RNA for qRT-PCR experiments and proteins for immunoblots.Designing target primers for qRT-PCR experiments and designing antibodies for immunoblot experiments.Successfully executing qRT-PCR and immunoblot experiments.Success indicator: Identification of RNA's and transporters associated with border cell metabolite secretion transporters that are upregulated under biotic elicitation conditions.Aim 3:Cultures of S. melilot and P. syringae DC3000 will be acquired.Culturing S. melilot and P. syringae DC3000 in their appropriate medias and retainment of glycerol stocks for future use.Execution of growth inhibition assays.Successful completion of the growth inhibition assays will be recognized after a minimum of three identified border cell secreted metabolites have been utilized in growth inhibition assays.Success indicator: Identification of metabolite-based growth inhibitors or promoters.EffortsUndergraduate MentoringThe University of Missouri-Columbia has a strong, established undergraduate presence in research laboratories across campus. We will recruit new and continue to train current undergraduate students throughout the duration of this award. Further details are outlined in the Target Audience portion of this report. Rural High School Student OutreachPD will engage in rural high school outreach to education students on STEM careers. Further details are outlined in the Target Audience portion of this report.WorkshopsThe Sumner laboratory hosts an annual metabolomics workshop that is available to members of the University of Missouri-Columbia. The PD will act as a teacher in this workshop both years of the project award. Further details are outlined in the Target Audience portion of this report. ConferencesPD will actively participate in conferences throughout this award. This will include electing to give oral presentation or, if not selected for oral presentations, poster presentations. Further details are outlined in the Target Audience portion of this report.

Progress 07/01/23 to 06/30/24

Outputs
Target Audience: Rural Highschool Students Progress: PD has joined Science on Wheels at the University of Missouri-Columbia. Thus far, Science on Wheels has not provided any experiences reaching out to rural high school students due to lack of funding. As Science on Wheels historically works with rural high school students, PD has been in conversations with Science on Wheels organizers to obtain funding for rural high school outreach. This is an ongoing passion for the PD (who is from a rural area) to educate rural high school students on advanced careers in STEM. Undergraduate Students, University of Missouri-Columbia Progress: PD has actively engaged with mentoring two undergraduate students. The first is an active student in the PD host laboratory (Lloyd Sumner laboratory, Univ. of Missouri). Now a sophomore at Univ. of Missouri, this student is a woman from Brazil that PD has taught basic laboratory skills and aided in advancing her understanding of metabolomics/mass spectrometry. This student is exceptionally high achieving, having been awarded two undergraduate research fellowships and currently undertaking a DAAD Research Internship in Science and Engineering in Germany. The second is a senior undergraduate man from a laboratory in mechanical engineering, in which PD taught basic plant science skills. This student has since graduated from Univ. of Missouri and will be beginning a PhD program at Univ. of Missouri in Fall 2024. University of Missouri-Columbia Metabolomics Center Annual Workshop Participants Progress: PD has actively taught in one Univ. of Missouri-Columbia metabolomics core sponsored workshop. This was a once yearly weeklong session, with a minimum of five hours of laboratory time each day. Participants were taught basic metabolomics workflows, starting sample extraction to derivatizations suitable for LCMS and GCMS. This also included introduction to instrumentation and data processing. Attendees included primarily graduate students and postdoctoral fellows. The backgrounds of attendees were a diverse mix of cultures and backgrounds including underrepresented minorities. Approximately half of the training group were women. Several attendees were from other universities in the Univ. of Missouri system, broadening PD efforts in audience reached. Conference Attendees Progress: Univ. of Missouri College of Agriculture, Food, and Natural Resources (CAFNR) Research Symposium: The CAFNR research symposium is a Univ. of Missouri campus wide event. As such, it was widely multi-disciplinary, including participants from both STEM and non-STEM backgrounds. Furthermore, participants included undergraduate and graduate students in addition to postdoctoral fellows. PD presented a poster and spoke with a wide variety of attendees, many of non-plant science backgrounds. Metabolomics Association of North America (October 2023): The Metabolomics Association of North America consists of a select audience of several hundred individuals, primarily from a mass spectrometry/metabolomics background. As an international conference, attendees are from diverse backgrounds and universities spanning primarily Mexico, Canada, and the United States. American Society of Mass Spectrometry (June 2024): The American Society of Mass Spectrometry is the premier conference for scientists utilizing mass spectrometry. As a massive international conference (ASMS 2024 was between 6-7,000 attendees from around the world) featuring academic and industry researchers, the target audience was from a broad background. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Training: Mass Spectrometry Instrumentation: PD has gained valuable experience using trapped ion mobility spectrometry (tims) through the Sumner lab tims-tof instrument. The Sumner lab has also recently acquired an orbitrap (Thermo Exploris 480) on which PD has now been trained. Through both of these instruments, PD has gained considerable knowledge on mass spectrometry instrumentation. These expertise will prove valuable as PD moves forward with research outlined in this project and in future career endeavors. RNA extraction, qRT-PCR workflows, and instrumentation: PD has gained knowledge of necessary procedures for plant RNA extraction, quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) workflows, and associated instrumentation for qRT-PCR. This enables PD to perform multi-faceted experiments, wherein traditional molecular biology approaches like qRT-PCR can be combined with cutting-edge mass spectrometry instrumentation. This is a powerful approach, effectively allowing PD to independently perform advanced experiments that possess greater research impact. Ultimately, especially considering the knowledge gaps in root border cell research, this allows greater progress/impact in the field of border cell research. Professional Development: Bayer-University Mentorship Program (B4U): PD has now completed B4U, which is a competitive annual program that provides one on one mentoring for students and post-doctoral researchers. This mentee driven program offered mentees with soft skill coaching and career development advice. Each mentee was paired with an industry scientist at Bayer as a mentor to generate professional development goals for career development, and they worked together to complete those goals. Bayer provided structured content on soft skill development, an understanding of industry working environment, and expectations of an influential scientist. Current Co-coordinator Univ. of Missouri Interdisciplinary Plant Group (IPG) Plant Talks: IPG Plant Talks is a three times per month seminar that takes place in the Fall and Spring semesters each academic year. This program is student/post-doc lead, in which two coordinators (which include PD and one graduate student in Univ. of Missouri Plant Sciences) arrange undergraduate/graduate student and post-doc speakers. Plant Talks also arranges two industry-based representatives each semester to come and discuss their careers in industry as a form of networking and soft skill development.PD would like to mention a personal enjoyment in serving as co-coordinator, as it has proven very rewarding to arrange research presentations from student/postdoc speakers on the Univ. of Missouri campus. Conferences: PD has attended three conferences over the 2023-2024 award year. These consist of the Metabolomics Association of North America (MANA) 2023, Mizzou College of Agriculture, Food, and Natural Resources (CAFNR) Research Symposium 2023, and American Society of Mass Spectrometry 2024. PD was part of the local organizing committee and scientific organizing committee for MANA 2023. This taught PD valuable skills in organization, communication, team work, and evaluation of scientific merit. At MANA 2023, PD also provided an oral presentation, which encouraged PD public speaking skills, networking, and enhanced research dissemination. At the remaining conferences, PD presented a poster which also encouraged public speaking skills, networking, and enhancement of research dissemination. How have the results been disseminated to communities of interest?Research has been disseminated at three conferences as of this progress report to a wide, research-based community. Two conferences primarily contained metabolomics/mass spectrometry professionals. Our novel approach to plant-based metabolomics has been well received due to our combinatorial approach on hardware development and instrument method development for live tissue usage. While these conference featured attendees from a plant science background, the vast majority were from a medical background. As such, PD was able to reach an audience needing deeper understanding of the complexity of plant systems and therefore the necessity of plant science research. The last conference featured many different disciplines both STEM and non-STEM related. This allowed education of a broad community at different experience levels on border cell functions. Many attendees arose from the humanities, meaning PD had a unique experience explaining border cells to non-scientists. What do you plan to do during the next reporting period to accomplish the goals? Training: Goal 2: Goal 2 is currently in progress. To accomplish this goal, PD plans to continue receiving training as needed from aforementioned collaborators. This will also entail further education on experimental design. Goal 3: As this goal has not been started, PD will be working with collaborators identified on campus to establish an experimental protocol and receive training. PD has already received some introductory training, helping with quick entry to this goal. Goal 4: As this goal has not been started, PD plans to perform literature searches to learn proper methodologies in growth inhibition assays. As part of training, PD plans to conduct initial experiments using known compounds from established literature. Successful replication of these experiments should suffice for training and establish a firm basis for experimental treatments using identified metabolites from these studies. Research: Goal 2: PD has already started this goal and plans to further refine an experimental protocol. PD also plans to continue working with collaborators as necessary for experimental design/procedures. PD plans to work with collaborators on proper data presentation and transparency for publication. Goal 3: PD has not yet started on research goal 3. To meet this goal, PD plans to establish an experimental protocol and carry out immunoblot experiments. PD has established communications with a collaborator on-campus to help with teaching necessary experimental design/procedural skills and data presentation. PD plans to acquire necessary materials within the next quarter in preparation for these experiments. Goal 4: PD has not yet started on research goal 4. To meet this goal, PD plans to establish an experimental protocol and perform preliminary experiments to ensure consistency with established literature. After ensuring proper protocol has been established, PD will carry out experiments as outlined in project initiation. PD has already acquired the bacterial species necessary for these growth inhibition assays. As this goal is relatively straightforward, PD expects this goal to progress relatively quickly compared to previous goals.

Impacts
What was accomplished under these goals? Training Goal 1 has been completed with the following results: PD has received training from Bruker personnel in operation and management of the Sumner lab tims-tof instrument. Border cell secretions have been analyzed using UHPLC-tims-qToF-MS. Data on border cell secretions has been processed and some compounds have been identified. Identification of more metabolites is ongoing as of this progress update. Goal 2 has been completed in part and is currently ongoing in line with PD submitted timeline. Current progress is as follows: PD has reached out to collaborators for instruction in proper qRT-PCR usage and methods development. This training has been generously provided by Dr. Norma Castro-Guerrero, a scientist in the laboratory of Dr. David Mendoza Cozatl. A preliminary experimental protocol has been developed based on utilization of Medicago truncatula full root tissues. A derivative protocol has also been developed for utilization of M. truncatula root border cells, which need a slightly different approach due to tissue constraints. Data collection is currently in progress, with the next step being data assembly for publication. As such, after data is collected and analyzed, the next step will be the last milestone of Goal 2. Goals 3 and 4 are to be performed under the 2024-2025 project year. Research Goal 1 is completed as outlined in project initiation. Thus far, PD has achieved all milestones outlined below: Major issue: Root border cell secreted metabolites/compounds influence the surrounding soil but are at extremely low concentrations. Current high-end mass spectrometry instrumentation can detect these compounds, but a protocol for harvesting was needed. PD aimed to develop a protocol for harvesting and identifying border cell secretions. Research Aim 1 Milestone 1: A protocol for the collection of border cell secreted metabolites has been developed. This protocol maintains border cell viability and allows collection of border cell secretions in sufficient amounts for later mass spectrometry analysis. The developed protocol entails using a bioreactor chamber containing border cells. The bioreactor system was custom designed by PD and Lloyd W. Sumner (PD host mentor). A liquid flow is established through the bioreactor such that border cell secretions are washed away. These secretions are then trapped on a downstream Solid Phase Extraction device. Solid Phase extraction allows direct harvesting of compounds from solution, making it ideal for trapping border cell secreted compounds. Research Aim 1 Milestone 2: PD is currently performing more collections of border cell secreted metabolites. The goal of these further collections is to increase depth of coverage/identifications of secreted metabolites. However, this milestone has been completed as PD possesses finalized mass spectrometry data suitable for publication. Research Aim 1 Milestone 3: PD has identified newly secreted metabolites arising from isolated border cells. As mentioned in the milestone progress update above, PD is currently performing more secreted metabolite collections to increase depth of coverage in the pursuit of more compound identifications. Current evidence has shown a wide diversity in compound secretion, with most identified compounds possessing properties relating to plant defense or induction of symbiotic (beneficial) interactions. Research Aim 1 Milestone 4: Several high-level secretions have been identified that are available through commercial vendors. Key Outcomes/Impact: Key outcomes from Research Aim 1 were a custom system/method for border cell secreted metabolite collection/analysis and evidence of border cells secreting metabolites independently of the plant root. This will prompt action on further root border cell research in plant health studies, supporting development of robust crop species. This ultimately will impact food quality and quantity for human consumption. We expect this research to be of interest to academic and private industry researchers in the near future. However, we expect longer term impacts to entail economic benefits for farmers and health benefits for the general public. Goal 2 is currently in progress, in line with the overall timeline provided upon application for this postdoctoral fellowship. Current progress on Goal 2 is outlined as follows: A protocol for collection of border cell RNA has been developed and is currently being refined as needed for optimal RNA yield. qRT-PCR materials have been acquired and some preliminary experiments have been performed in M. truncatula total root tissues as proof of concept. These have been performed successfully and PD is confident in future success using border cells in qRT-PCR. Goals 3 and 4 are to be performed under the 2024-2025 project year.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Kranawetter, C. and Sumner, L.W. (2024, 4 June). Poster Presentation. Preliminary Identification of Specialized Metabolite Secretions and Secretory Pathways in Medicago truncatula Border Cells. 2024 American Society of Mass Spectrometry (ASMS) conference (international conference).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Kranawetter, C. and Sumner, L.W. (2023, 6 October). Poster Presentation. Metabolomics of Secreted Specialized Metabolites from Isolated Medicago truncatula Border Cells. 2023 University of Missouri-Columbia College of Agriculture, Food, and Natural Resources (CAFNR) Research Symposium (local symposium).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Kranawetter, C. and Sumner, L. W. (2023, 26 October). Oral Presentation. Natural Product Secretion Patterns and Mechanisms of Isolated Border Cells from Medicago truncatula. 2023 Metabolomics Association of North America (MANA) (International Conference). Columbia, MO.