Source: UNIVERSITY OF FLORIDA submitted to
INVESTIGATING THE UTILITY OF FERAL POPULATIONS FOR BRASSICA CROP BREEDING
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1030769
Grant No.
2023-67012-40241
Cumulative Award Amt.
$169,300.00
Proposal No.
2022-09745
Multistate No.
(N/A)
Project Start Date
Aug 1, 2023
Project End Date
Jul 31, 2025
Grant Year
2023
Program Code
[A1141]- Plant Health and Production and Plant Products: Plant Breeding for Agricultural Production
Project Director
Mabry, M. E.
Recipient Organization
UNIVERSITY OF FLORIDA
G022 MCCARTY HALL
GAINESVILLE,FL 32611
Performing Department
(N/A)
Non Technical Summary
As climate change and other downstream impacts of human activities continue to cause increased drought and temperature stress, it is essential to find new ways to ensure food security. One innovative approach is to study how feral crops - plants that have escaped from cultivation and successfully established - respond to climate change and compare their genetic makeup to cultivated crops. This information can help identify populations of feral crops that can be sampled and used to supplement existing germplasm collections as well as to identify useful candidate genes for future food production. Feral populations may have unique adaptations to various climatic challenges, such as heat stress or soil salinity, that can benefit food production. This project proposes to use herbarium collections to inform and improve future food security by identifying feral crop populations for germplasm collection and conservation within the National Plant Germplasm System. This approach will highlight the potential of herbaria as a valuable resource for both protecting biodiversity and improving crops in novel ways. By identifying and conserving feral populations of crops, we can tap into untapped genetic diversity and ensure food security in the face of climate change.
Animal Health Component
5%
Research Effort Categories
Basic
95%
Applied
5%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2011440108050%
2021440106010%
2061440107020%
2131440106010%
9031440106010%
Goals / Objectives
The PD's overarching career goal is to become a leading agricultural science researcher at a Research-2 university while concomitantly being an excellent teacher and mentor, affording a non-traditional pool of undergraduate students with unique training opportunities. The proposed project aims to expand our understanding of feralization in vegetable crops, specifically Brassica oleracea and B. rapa, to determine the potential of feral crop populations as sources of genetic diversity for combating persistent and increasing threats to food security, such as climate change. The project aims to investigate the frequency and timing, genetic architecture, potential breeding applications, and environmental conditions that drive feralization. To achieve these goals, the project will utilize natural history collections, ecological niche modeling, artificial intelligence methods, and genomics to identify the environmental and genomic signatures of ferality. The results and materials created will act as starting resources for the PD's own research lab and will ultimately contribute to new ways of thinking about paths to innovation for future food security.Objectives - The PD will devote full-time effort (100%) to the project during the course of the fellowship.1. Defining the ecological niche of feral Brassica oleracea and B. rapa.1.1 - Identification of feral populations of crop species.1.2 - Development of ecological niche models.1.3 - Leveraging AI to capture traits not in environmental datasets.2. Comparative genomics of feralB. oleraceaandB. rapa.2.1 - Re-sequencing feral populations.2.2 - Frequency and pathways to ferality.2.3 - Identifying loci associated with feralization and local adaptation.
Project Methods
The methods described in Aim 1 of the research proposal involve defining the ecological niche of feral Brassica oleracea and B. rapa to better understand ferality and its implications for climate change adaptation. In aim 1.1, locality data from digitized herbarium specimens of B. oleracea and B. rapa will be filtered based on known historic and current cultivation zones, including botanical gardens and farms. Cultivation zones will be determined using text searching of label data and Landsat imagery. In aim 1.2, environmental variables such as elevation, soil characteristics, land use, and bioclimatic variables will be assembled from herbarium records and other sources. Ecological niche models for feral B. oleracea and B. rapa will be produced using Maxent software, which uses machine learning algorithms to predict species distributions based on environmental variables. Models will be projected onto maps of current and future environmental conditions using WorldClim databases. In aim 1.3, AI methods will be used to detect changes in phenology (e.g., flowering and fruiting time) of feral populations using herbarium specimens. The ResNet50 convolutional neural network, which is pre-trained on millions of images, will be used to classify reproductive vs. non-reproductive specimens with high accuracy.The methods proposed in Aim 2 of the study include re-sequencing feral populations of B. oleracea and B. rapa, analyzing the frequency and pathways to ferality, and identifying loci associated with feralization and local adaptation. In Aim 2.1, leaf tissue will be collected from feral populations of B. oleracea and B. rapa across the US, and genomic DNA will be extracted and sequenced using whole genome re-sequencing at Rapid Genomics LLC. The sequencing will result in paired-end reads at a depth of 11x. The sequence data will then be aligned to reference genomes of B. oleracea and B. rapa using the Burrows-Wheeler Aligner (BWA) software, and single nucleotide polymorphisms (SNPs) and insertions/deletions will be called using the Genome Analysis Toolkit (GATK) software. All analyses will be conducted on the HiPerGator server maintained by the University of Florida Research Computing Facility. In Aim 2.2, the newly generated sequence data from feral populations will be combined with previous sequence data from cultivars and wild relatives to assess the relationships between different populations and the frequency of feralization. Two phylogenetic programs, SNPhylo and TreeMix, will assess individual sample relationships and introgression to determine the pathways to ferality (endoferal or exoferal). Four-population (f4) tests will also be used to test the support of inferred migration edges from Treemix. Ancestry proportions and genetic structure of populations will be assessed using fastSTRUCTURE, and population structure, allele frequencies, and individual inbreeding coefficients will be evaluated using PCAangsd. In Aim 2.3, loci associated with feralization and local adaptation will be identified. Differentiation outliers and genetic-environment association tests will be used to identify loci with high differentiation between populations. FST methods will be used to calculate genetic differentiation, and the Bayenv program will be used to analyze correlation tests between allele frequencies and the environmental variables which best describe the niche of feral crops as identified in Aim 1. Variants identified in both methods will be further investigated as potentially valuable for targeted modifications enabling crops to be more locally adapted. Additionally, known genes that regulate leaf size, flowering time, and pathogen resistance will be analyzed using genome-wide association studies (GWAS) to identify potential candidate genes associated with feralization and local adaptation.The main advisor group will be primary mentor Dr. Pam Soltis and collaborating mentors Drs. Doug Soltis and Alex McAlvay. They will assess and evaluate the quality of research and professional development. They will critique the expected outcomes by preparing manuscripts and presentations. The rationale for their role is to ensure that robust and reproducible science is being conducted and all data produced will be stored using FAIR principles (Findable, Accessible, Interoperable, Re-Usable) and rooted in CARE (Collective Benefit, Authority to Control, Responsibility, and Ethics) best practices. Beyond the project mentors, Dr. Lucas Majure (curator of the Florida Museum of Natural History Herbarium) and Dr. Zachary Stansell (curator of the PGRU Vegetable and Hemp Crop Collections) will act as an additional advisory group. The rationale for their role is to ensure that herbarium vouchers and germplasm are deposited promptly and with all associated metadata for long-term storage and community use. Peers at the Florida Museum of Natural History, University of Florida, and New York Botanical Garden will also act as a supplementary advisory group. Presentations at internal seminars will allow for feedback on the proposed research's quality, expected measurable outcomes, and impacts. The rationale for their role is for feedback from different experts. Finally, to ensure that the research is being evaluated by different types of expertise and experience, attendees at national meetings (National Association of Plant Breeders and Plant and Animal Genome) and the Project Directors meeting and Foundational Area meeting are other opportunities for evaluation of the proposed research. The attendees at the meetings are composed of peers and government officials who will give different forms of feedback on the research. Finally, attendance at the Biology and Mathematics Educators (BIOME) Institute will serve as a way to assess and evaluate the quality of the PD's teaching strategies and diversity and inclusion efforts.

Progress 08/01/23 to 07/31/24

Outputs
Target Audience:In this reporting period, the project targeted academic and scientific researchers in agricultural genetics, ecology, and evolutionary biology who are interested in understanding feral crop populations, especially those within Brassica. Efforts were made to connect with researchers and naturalists in relevant regions to coordinate sampling and permit processes, advancing project objectives and building a collaborative foundation for analyzing feral Brassica populations. Additionally, undergraduate students at the University of Florida were targeted through hands-on research experiences, engaging them in data collection and filtering efforts using iDigBio and GBIF databases. This included analyzing occurrence records of B. oleracea and B. rapa and filtering records based on historic and current cultivation zones, employing land use maps, and filtering records by keywords associated with ferality. These activities aim to foster interest in crop conservation and ecological genetics while training future researchers in practical and computational methods for biodiversity studies. Changes/Problems:During this reporting period, the project encountered some logistical delays, though these were managed to minimize the impact on overall progress and project goals: Field Sampling and Permit Delays: Coordinating field collections of feral Brassica populations required additional time to secure permits and organize collaboration with local researchers and naturalists. Although this caused slight delays in sample collection, these partnerships have ultimately strengthened the project's regional network and should streamline future sampling efforts. Data Management and Filtering Complexity: Analyzing herbarium data from iDigBio and GBIF and filtering it to identify feral populations based on cultivation zones introduced unanticipated challenges due to the volume and variability of the data. To address this, additional time was devoted to refining the data filtering criteria and training the undergraduate researcher. This adjustment may impact the timeline for downstream genomic analyses; however, the improved dataset will enhance the quality of analysis and ensure a comprehensive examination of feral populations. Adjustment to Data Management Plan: No major changes were made to the Data Management Plan itself, but the approach to compiling and verifying the dataset required modifications to accommodate more detailed filtering and ground-truthing of herbarium records. These adjustments have made the database more robust, supporting future publications and data sharing. Despite these adjustments, no significant deviations from the research schedule or goals are anticipated beyond minor timeline extensions for sample collection and dataset preparation. The project remains on track to meet its objectives of re-sequencing feral populations and identifying new data patterns within Brassica diversity. There were no changes in approved protocols. What opportunities for training and professional development has the project provided?Undergraduate Mentorship: The project provided an undergraduate student with training in biodiversity data analysis, using databases such as iDigBio and GBIF for identifying and filtering herbarium records. This experience introduced the student to practical skills in ecological and genetic data handling and basic geospatial analysis using land use maps, enhancing their skill set in biodiversity informatics and data science. Field Collection and Sample Management: Engaging with local researchers and naturalists offered the project team--including the student researcher--practical experience in field sampling, species identification, and coordinating fieldwork logistics. This experience contributes to their professional development in ecological fieldwork and permits coordination. How have the results been disseminated to communities of interest?Presentation at CROPS Conference: Preliminary findings and methods were presented at the CROPS conference, reaching an audience of agricultural researchers and professionals interested in crop biodiversity and conservation. This venue enabled knowledge exchange and collaboration with other researchers focused on genetic diversity and feral populations. Collaborative Outreach with Local Naturalists and Researchers: Through partnerships with naturalists and researchers in various U.S. regions, the project engaged with individuals directly involved in local biodiversity efforts. This outreach has helped expand the network of stakeholders interested in feral Brassica populations and their potential for informing biodiversity conservation. What do you plan to do during the next reporting period to accomplish the goals?Expand Sample Collection: Additional field sampling of Brassica populations will continue to increase the genetic diversity represented in the project's physical collections, aiming to include populations from diverse cultivation zones. Genomic Analysis: Begin the genomic sequencing of the collected samples to analyze genetic diversity and determine the relationship between feral, cultivated, and wild populations of B. oleracea and B. rapa. The sequencing results will support publications and contribute new data to genetic studies on feral populations. Data Refinement and Analysis: Further refine and analyze the compiled dataset to identify trends in feral population distribution and characteristics across U.S. regions, incorporating additional ecological and land use data. This analysis will guide the selection of populations for sequencing and contribute to forthcoming research articles. Present Findings at Conferences and Publish Initial Results: Share progress through additional conference presentations and aim to prepare initial publications to disseminate findings to the academic community.

Impacts
What was accomplished under these goals? During this reporting period, the project made progress under two primary goals: Goal 1: Identification of Feral Populations - An undergraduate researcher worked to identify potential feral populations of Brassica oleracea and Brassica rapa using data from iDigBio and GBIF. Locality data were filtered based on known cultivation zones and land use patterns, with keywords associated with ferality (e.g., "escaped") used to refine the dataset. The resulting database provides a robust foundation for further analysis of potential feral populations in the U.S. Goal 2: Re-sequencing of Feral Populations - Feral populations were sampled from various U.S. regions. This included coordinating with local researchers and naturalists to collect Brassica samples and securing necessary permits, which enabled us to create a physical collection for genomic re-sequencing. These samples will be analyzed to compare genetic diversity among feral, wild, and cultivated populations of Brassica species, filling a gap in current genetic datasets.

Publications