Source: UNIV OF MINNESOTA submitted to NRP
THE EFFECTS OF SEASONALITY AND CALLOSE DEPOSITION ON CARBON TRANSPORT IN CROP PLANTS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1030581
Grant No.
2023-67012-40180
Cumulative Award Amt.
$186,000.00
Proposal No.
2022-09727
Multistate No.
(N/A)
Project Start Date
Jul 1, 2023
Project End Date
Jun 30, 2025
Grant Year
2023
Program Code
[A1152]- Physiology of Agricultural Plants
Recipient Organization
UNIV OF MINNESOTA
1035 UNIVERSITY DR
DULUTH,MN 55812-3011
Performing Department
(N/A)
Non Technical Summary
Drought, frost, and other stresses can reduce plant productivity and crop yields. Sugar movement around the plant body to areas of growth and development drives productivity. The timing of stress, may affects current-season productivity. Plant stresses stop or slow sugar movement and plant growth. It is unknown if stress timing can reduce productivity in future growing seasons. This project will determine if the timing of drought or other stresses can reduce sugar movement and thus plant productivity. Does a drought that occurs during summer limit a tree from making fruits or flowers the following spring?We will grow trees in controlled greenhouse conditions and expose them to drought at different times during the year. After each stress treatment, we will make measurements of plant health, ability to make sugar, and ability to move sugar. We will continue making measurements each season for one year. The extra year of measurements will allow us to determine when the trees are able to recover from the stress. The broad implications from this work will be more reliable crop production. Specifically, our data will allow those who grow crop trees to better react to droughts and other stresses. Further, this work will allow stakeholders to better predict yields after plant stress.
Animal Health Component
0%
Research Effort Categories
Basic
100%
Applied
0%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
20624991020100%
Knowledge Area
206 - Basic Plant Biology;

Subject Of Investigation
2499 - Plant research, general;

Field Of Science
1020 - Physiology;
Goals / Objectives
1. Conduct a two seasonal studiesof callose deposition after drought and delayed winter dormancy.a. Determine turgor loss points for the target species.b. Measure chlorophyll fluorescence, callose presence, callose location, phloem anatomy, leaf stomatal conductance, leaf photosynthetic rate on the target species once per season.c. record phenological stages for the greenhouse population and outdoor conspecifics seasonally: noting the timing of leaf bud break, floral bud break, open leaves, open flowers, yellow leaves, and leave abcission.c. In the second year, also measure phloem sap flow rate for target species during each seasonal timepoint.2. Analyze the data from each study3. Prepare manuscripts describing the outcome of each study
Project Methods
We will purchase and maintain plants in a greenhouse setting, monitoring them for proper hydration, nutrition, and pests.Proior to beginning the seasonal measurements, plant turgor loss point will be found using a pressure chamber and drought treatment.Seasonally, we will measure:Chlorophyll fluorescence with a fluorometer and with a LiCor portable photosynthesis systemThe amount of callose in the phloem via fluorescence assayThe location of callose and amount of callose occlusion on the sieve plates with scanning electron microscopySieve element and phloem anatomical characteristics using classical histological sectioning with a sliding microtome and staining with Safranin O and Alcian Blue. These measurements will be used to calculate the leafLeaf Stomatal Conductance and leaf Photosynthetic Rate with a LiCor portable photosynthesis system.In year 2 we will also measure the flow rate of the sap within the sieve elements using phloem-mobile dyes such as CFDA or Esculin.We will analyze the results using regressions, t-tests, and ANOVA, to determine differences between phenological stages between and within species.All measurements will provide experiential learning opportunities for an undergraduate student.Evaluation of project success will be determined by:1) Completing all measurements as planned, or completing suitable alternatives in cases where methods do not work.2) Compiling results, analyzing them, and writing them into two manuscripts submitted to peer-reviewed journals.3) Presenting intermediate and final results at two academic conferences.

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

Outputs
Target Audience:Scientists performing plant biology work, both applied and basic. Reached through informal contact in meetings and at a major national meeting (American Society of Plant Biology). Changes/Problems:After consulation with an expert in Citrus biology, the inclusion of citrus in the experiment was altered. The broad concern was that growing the plants in Minnesota with very large changes in day length could alter the "normal" physiology of the plant. It was removed from the seasonal drought experiment as described in the proposal and has been includedin a new sub-project focused citrus seasonality. The project will investigate the changes to carbon movment ability in different flushes of citrus branches in plants infected and not infected with HLB. The overall goal of the project remails to undersand the structure-function relationships in seasonal phloem phenology with specific emphasis on callose deposition as described in the proposal. The material is being collected by researchers at the University of Florida Research Station and will be shipped to Minnesota where I will process and analyze it. This project will result in its own manuscript. What opportunities for training and professional development has the project provided?1) I have learned how to digest samples for callose and structural analysis by SEM, a technique that is generally underused for phloem analysis. 2) Experiencedirectly managing grant budgets and major grants including interfacing with University Sponsored Projects office. 3) Mentored one student directly as a PI. How have the results been disseminated to communities of interest?At a national conference (American Society of Plant Biologists) and through international, discipline specific journal club. What do you plan to do during the next reporting period to accomplish the goals?The final drought experiment will be conducted late July through August 2024. Data for all droughts will be analyzed, summarized and prepared as a manuscript for submission in a peer-reviewed journal. Citrus material will be sectioned, analyzed and the resulting data will be analyzed as it is available. Scanning electron microscopy will be conducted at the Arnold Arboretum at Harvard University through a number of trips to Massachusetts and the resulting data will be analyzed, summarized and included in the appropriate manuscript.

Impacts
What was accomplished under these goals? 1) The first seasonal study is underway and on target to be completed in August 2024. For this study: turgor loss points were determined and for the seasonal study periods that have already been completed (fall and spring), all physiological measurements were taken. Callose presence and location is currently being investigated with Scanning Electron Microscopy. Phenological periods are being monitored and recorded as they occur. Followup measurements for the fall drought were conducted in May 2024. The second seasonal study has been altered due to concerns over growing citrus under the daylight regime in Duluth (specifically short winter days). The material is currently being collected at the University of Florida Citrus Research Station. 2) Data for fall was analyzed and data from spring is currently being analyzed as the drought experiment ended in late May. Data from the spring followup study has also been analyzed. 3) Manuscript preparation has not started as the experiment is not complete.

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