Progress 05/25/21 to 06/01/23
Outputs
Target Audience:The target audience: Blueberry breeders, farmers, molecular geneticists, and other allied scientists of this crop, consumers and the general public. Production of blueberry fruits greatly benefits from insect-mediated pollination, for which it is becoming common practice among growers to supplement pollination services with managed honeybees. By comparing flower and nectar rewards and pollinator preferences for the flowers, this project aims to identify the cultivars preferred by pollinators and consequently visited and pollinated more frequently. This project seeks to understand why this is happening at a molecular level and provide guidance to farmers on the cultivars preferred from a pollinator perspective, hence potentially more productive. The outcomes of this project will allow blueberry producers to give value to their products and, at the same time, respond to the requests of consumers who are increasingly responsive to fruits obtained from organic and environmentally friendly orchards. Changes/Problems:The project had a delayed starting date as of September 2021 because of the difficulties in hiring a postdoctoral researcher, who left the country after three months (in December 2021) because of family issues. A second postdoc was hired to work on the project in August 2022, and he has been working on this project since then. We encountered difficulties extracting RNA of sufficient quality and quantifying for the gene expression experiments that were solved at the beginning of 2023. What opportunities for training and professional development has the project provided?One postdoc has been trained on this project on the following analysis: analysis and visualization of metabolites, extraction of RNA and analysis of RNAseq data. How have the results been disseminated to communities of interest?The results of this project have been presented to farmers, breeders, stakeholders, and the scientific community in the form of oral presentations and posters, including: the Washington Blueberry Research Review, Mount Vernon, WA (October 2021); Washington Small Fruit Conference, Lynden, WA (December 2021); Hansen Retreat, Utah State University (October 2022); University of Florida, Dept. of Horticulture, internal seminar (December 2021); Annual Meeting of the Phytochemical Society of North America (July 2022); Annual meeting of the Washington Blueberry Commission (October 2022). Dr. Borghi and Dr. DeVetter wrote a commentary about blueberry nectar published in the Small Fruit Update of the Northwestern Berry Foundation (May 2023). Dr. Borghi also included a module on the chemical signals for pollinators' attraction and rewards, hence nectar and pollen, in the specialty course BIOL6750 she teaches every spring semester at Utah State University. What do you plan to do during the next reporting period to accomplish the goals?We are working on RNAseq data analysis and interpretation to compile a comprehensive manuscript on blueberry floral chemical traits of pollinator attraction and reward, including the genes for metabolite synthesis.
Impacts
What was accomplished under these goals?
We completed the analysis of nectar and pollen metabolomics of four blueberry cultivars, namely Calypso, Draper, Liberty, and Last Call, collected from Washington State University experimental fields. In parallel, we performed greenhouse experiments to assess the morphological parameters of blueberry flowers (size, corolla width, pistil and anther length, etc). Pollinator counts in the field have also been completed and correlated with data on fruit size and yield. Because of the difficulties in hiring a postdoc to work on this project, metabolomics assessments of flowers exposed to heat stress could not be performed. However, we extended the work on the cultivars and performed RNAseq of flowers. In collaboration with Dr. Gutesohn (West Virginia University) we collected, quantified and annotated volatile organic compounds (VOCs) emitted from flowers. In collaboration with Dr. Schaeffer (Utah State University), we performed multiple choice assays with bumblebee pollinators to assess pollinator preferences for VOCs emitted by different flowers.
Publications
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2021
Citation:
Presentations:
Borghi, M., UF, Dept. of Horticulture (UF-IFAS), Invited Seminar, "High-energy level metabolism and transport reveled at the transition from close to open flowers.," University of Florida, Dept. of Horticulture, UF-IFAS. (December 6, 2021)
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2021
Citation:
Presentations:
DeVetter, L. (Presenter & Author), Borghi, M., Washington Small Fruit Conference, "Pollination Update-Floral Traits," Lynden, WA. (December 1, 2021)
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2021
Citation:
Presentations:
DeVetter, L. (Presenter & Author), Borghi, M., Washington Blueberry Research Review, "Pollinator attraction - Nectar, pollen, and assessment of new technologies," WA Blueberry Commission, Mount Vernon, WA. (October 14, 2021)
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Progress 10/01/21 to 09/30/22
Outputs
Target Audience:Blueberry farmers, breeders, beekeepers. Changes/Problems:The postdoc initially assigned to this project left USU after 2.5 months of work. A new postdoc was hired in August 2022. What opportunities for training and professional development has the project provided?One postdoctoral researcher has been assigned to work on this project (PI start-up funds). The postdoc has been trained on RNA extraction and qRT-PCR How have the results been disseminated to communities of interest?The results of this research were presented at the annual meeting of the Washington Blueberry commission in October 2022. The postdoc currently working on this project presented preliminary results of differential gene expression at the USU Hansen retreat (Oct 2022) What do you plan to do during the next reporting period to accomplish the goals?RNA extractions of all samples are under way. VOCs will be collected starting in February/March, annotated and quantified.
Impacts
What was accomplished under these goals?
Nectar production differed among cultivars, and cultivars with less nectar had the lowest number of observed honey bee foragers. Regression analysis showed a positive but non-significant relationship between nectar production and honey bee forager counts, which is likely because pollinator recruitment is also influenced by floral morphology, a trait that differed significantly among cultivars. Although the total content of carbohydrates is not significantly different among cultivars, significant differences were instead observed at the level of individual carbohydrates. Total content of amino acids was also different among cultivars as well as the content of organic acids. Compounds with biological activity, such as nicotinate (also known as niacin or vitamin B3) and quinic acid were present in low abundance in nectar of two cultivars.
Publications
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Progress 05/25/21 to 09/30/21
Outputs
Target Audience:Blueberry breeders, growers, and nurseries; Beekeepers. Changes/Problems:We had difficulties in extracting a sufficient amount of RNA from blueberry flowers with conventional extraction kits which are commonly utilized to extract RNA from model plant material. To overcome these problems, we successfully contacted scientists who routinely work with blueberry samples and are in the process of purchasing reagents needed to complete RNA extractions. What opportunities for training and professional development has the project provided?One postdoctoral researcher has been assigned to work on this project (PI start-up funds). The postdoc has been trained on data analysis and visualization How have the results been disseminated to communities of interest?The preliminary results of this research have been presented at the following audiences: DeVetter, L.W., M. Borghi, G. LaHue, A. De La Luz, T. Sade. 2021. Pollinator attraction - Nectar, pollen, and assessment of new technologies. Washington Blueberry Research Review. Mount Vernon, WA. Oct. 14, 2021. DeVetter, L.W. and Borghi, M. 2021. Pollination Update-Floral Traits. Washington Small Fruit Conference, Lynden, WA. Dec. 1, 2021. Borghi, M. High-energy level metabolism and transport reveled at the transition from close to open flowers. University of Florida, Dept. of Horticulture, UF-IFAS (internal departmental seminar on Zoom). Dec. 6, 2021 What do you plan to do during the next reporting period to accomplish the goals?Pollen data will be annotated starting in January 2022. In the meantime, we started correlation and regression analyses between metabolites, pollinator visitation, fruit sets, and berry weights. These data will allow us to better understand the role that nectar and pollen metabolome plays in linking pollinator preferences with traits of relevance for the growers and end consumers. We are in the process of extracting RNA from flowers harvested from the same plants from which nectar and pollen were collected. Dr. Iorizzo (North Carolina State University) gave us personal access to the recent version of the V. corymbosumgenome sequence that we will utilize as a scaffold for RNA sequencing of our samples. Concurrently, we are vernalizing potted plants in the USU Greenhouse. These plants will be utilized to assess flower anatomy and structures for which potted plants grown under controlled conditions are preferred.
Impacts
What was accomplished under these goals?
Nectar and pollen from 4 highbush blueberries (Vaccinium corymbosum) cultivars Calypso, Draper, Liberty, and Last Call were collected from Washington State University experimental fields by the research group of Dr. Lisa DeVetter. Floral metabolites were extracted, derivatized, and separated using a Gas Chromatography-Mass Spectrometry (GC-MS), annotated and quantified by comparison with peaks and spectra of analytical standards. Pollen data are still being annotated and will be ready by the time of the next year report. Based on the nectar metabolite concentrations, blueberry cultivars cluster into two separate groups: 'Liberty' with 'Draper' and Calypso' with 'Last Call'. 'Last Call' has the highest sugar content (fructose, glucose, sucrose, and maltose) in its nectar despite low volumes, which is likely due to a concentration effect. Correlation and regression analyses between metabolites, pollinator visitation, fruit sets, and berry weights are currently underway.
Publications
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