Source: PENNSYLVANIA STATE UNIVERSITY submitted to NRP
ROOT SYSTEM FUNCTION AND NUTRIENT USE IN CEREALS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1021929
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Jan 1, 2020
Project End Date
Dec 31, 2024
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
PENNSYLVANIA STATE UNIVERSITY
208 MUELLER LABORATORY
UNIVERSITY PARK,PA 16802
Performing Department
Plant Science
Non Technical Summary
Global agriculture faces the complex challenge of delivering more for less: population growth will increase demand; current rates of fertilizer and agrochemical use will not be sustainable as limited natural resources become exhausted and the environmental costs associated with their use continue to rise; climate change will reduce both the area of high quality agricultural land and the length of the useful growing season. Although these are daunting prospects, there is every reason to be optimistic that significant improvements can be made to our agricultural systems, including enhancing the efficiency of the crop plants themselves. The broader crop gene pool (traditional varieties, crop wild-relatives, orphan and under-utilized crop species) is adapted to a wide range of resource-poor environments and low-input management systems. To date, mainstream breeding efforts have focused on high-input, high-density systems, leaving much of this potential untapped. The emergence of agriculture and the domestication of plant and animal species are among the defining events of the last 10,000 years of human history. The latest genomics technologies have the capacity to generate an unprecedented level of understanding of these processes and the rich diversity of crop varieties and agronomic practices present in the world today. Such understanding will be vital in our efforts to generate more efficient crops and cropping systems for a sustainable future agriculture.The root system is the primary connection of the plant to the environment and fundamental to the efficient use of water and soil nutrients. This project will further efforts to generate resource efficient crops by advancing our knowledge of root system function in maize (corn) and sorghum. Maize is the cornerstone of US agriculture, providing raw material in the form of starch, oil, protein and biofuel, while supporting further agricultural activities such as dairy and beef industries. Sorghum is a close relative of maize that is of African origin and displays good tolerance to heat, drought and nutrient-poor soils, and that is finding increasing importance in US agriculture. Across the global pool of maize and sorghum varieties (modern breeding material, traditional landrace varieties, and wild-relatives), root systems are highly diverse - in architecture and anatomy, in the molecular machinery of resource acquisition, in the compounds they secrete to the environment, and in their symbiotic interactions with other organisms. Using novel genomics data and custom generated genetic-mapping resources, this project will explore the genetic basis of this functional diversity and evaluate the ecological implications of the root system for plant adaptation to different soil and resource environments. Within the context of an internationally competitive academic research program, this project will generate resources, data and publications available to the research community, while providing extensive opportunities for undergraduate, graduate and postgraduate training. This project will fall within the wider strategic aims of the PSU Huck Plant Institute Root Biology Center collaborative network.
Animal Health Component
30%
Research Effort Categories
Basic
70%
Applied
30%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2011510108050%
2011520108050%
Goals / Objectives
1. Estimation of genotype-trait associations for root traits in maize and sorghum; generation of predictive polygenic models a) Curation of existing trait and genotype datasets b) Generation of novel sorghum root architecture/anatomy data c) Generation of novel maize/sorghum mycorrhiza response data 2. Exploration of genotype-environment/root-environment associations a) Application of polygenic trait prediction models to global maize/sorghum diversity b) Identification of environment/root trait correlations 3. Evaluate models in context of i) landrace/wild-relative diversity; ii) diverse agronomic scenarios a) Characterization of novel maize landrace-derived inbred families4. Mechanistic study of selected candidate variation a) Functional analysis of the Pho1 family of phosphorus transporters/sensors in maize and sorghum
Project Methods
1. Estimation of genotype-trait associations for root traits in maize and sorghum. Generation of predictive polygenic models.a) Curation of existing trait and genotype datasets (primarily maize root architecture/anatomy)We will collate and link pre-selected phenotypic, environmental and genomic data sets (Genetic datasets:SEED landrace GBS (maize)(Romero Navarro et al., 2017);(Gates et al., 2019);Global resequencing (sorghum)(Lasky et al., 2015; Bellis et al., 2019);Wisconsin diversity panel (maize)(Mazaheri et al., 2019);Sorghum association panel (sorghum)(Shakoor et al., 2016);Andosol landrace maize WGS (Sawers and Rellan-Alvarez, unpublished). Funcrional datasets: Root architecture and anatomy (Lynch);Seed element composition (maize)(Fikas et al., 2019);Seed element composition (sorghum)(Shakoor et al., 2016);Mycorrhizal response (sorghum) Sawers et al., unpublished. Environmental datasets:Bioclimhttps://www.worldclim.org/bioclim;Phenology-weighted climatic variables (maize)(Gates et al., 2019);Accession-linked climatic variables (sorghum)(Lasky et al., 2015; Bellis et al., 2019);Global soil modelshttps://soilgrids.org;Scaled P-retention prediction doi.org/10.5281/zenodo.1233213).Beyond the benefits of hosting different types of data in the same place, we will anchor all genomic information to the same genome reference and same gene-model annotation. While we anticipate that much will be possible by cross-referencing, re-mapping, and imputation, there may be good reason to generate additional genetic data - for instance, where (re-)genotyping of a panel that has been used for phenotypic evaluation would facilitate extrapolation to larger population genetic data sets. We will look to facilitate collective study of the two crops, leveraging understanding of their shared history and the relationship between their genomes.b) Generation of novel sorghum root architecture/anatomy dataTo investigate environmental correlations with root anatomy, we will evaluate a well-defined sorghum association panel (Shakoor et al., 2016) using a high-throughput laser ablation tomography (LAT) platform developed by PSU researchers (Figure 2. Saengwilai et al., 2014; York et al., 2015). In collaboration with Stephen Kresovich (Clemson), we will perform root sampling at a site in North Carolina, during Summer 2020. On site, we will dig up a predetermined sample of plants and collect their roots (whorls 2-4) for laser ablation. We will investigate the relationship between environmental factors at the site of origin and root anatomy.c) Generation of novel maize/sorghum mycorrhiza response dataWe will evaluate mycorrhizal response in a well characterized maize association panel (Mazaheri et al., 2019) using a small-pot greenhouse-based system. Young plants will be evaluated for biomass production and mineral accumulation, with or without inoculation with mycorrhizal fungi. We will use both microscopy and molecular methods to estimate mycorrhizal colonization. We will address the hypothesis that more stressful environments promote stronger mutualism, looking to link mycorrhizal symbiosis with specific environments. We will explore the impact of domestication and improvement on mycorrhizal response.2. Exploration of genotype-environment/root-environment associations.a) Application of polygenic trait prediction models to global maize/sorghum diversityWe will use standard genome wide association scans to identify SNPs linked with root architecture/anatomical traits and mycorrhizal responsiveness, using both existing data and data generated inthis project. Polygenic models generated from well-characterized diversity panels will be used to extrapolate phenotypic predictions to broader genotyped maize and sorghum diversity.b) Isolation of environment/root trait correlationsWe will use explore trait correlations (e.g. the relationship between root anatomy and mycorrhizal symbiosis) and trait-environment correlations. We will cross-reference the results of whole genome scans of environment-genotype relationships with phenotypic trait hits. In addition to identification of single-gene candidates, we will explore global relationships between environmental DNA variant candidates and specific traits. This approach will reveal the potential adaptive significance of trait variation with respect to different environmental gradients (e.g. precipitation, temperature, soil chemistry).3. Evaluation of models in context of i) landrace/wild-relative diversity; ii) diverse agronomic scenariosa) Characterization of novel maize landrace-derived inbred familiesOur approach requires the ability to infer root system functionality across global diversity using genetic models based on direct observation of relatively small samples.Heterogeneous, outbred landrace maize populations are difficult to work with and lack the genetic uniformity required for meaningful fine-scale evaluation. To address thisproblem, we are using conventional breeding techniques to introduce fragments of exotic maize diversity into a temperate inbred background. We willgenerate a collection of ~750 BC1S4 families derived from ~250 distinct donors, sourced from across the Americas. This material will be genotyped and evaluated during years 2-4 of this project. Evaluation will include both greenhouse and field components, taking in selected root traits, as well as an evaluation of mycorrhizal symbiosis, along with overall plant performance and ionomic profiling of both vegetative tissue and grain. Availability of this unique resource will be a key component of future external funding applications, and will provide opportunity for collaboration and impact beyond the immediate project.4. Mechanistic study of selected candidate variation.We will provide functional follow-up of a specific family of gene leads identified on the basis of prior knowledge in other plant systems and through the results of preliminary genotype-environment scans. It is anticipated that the activities in Goals 1-3 will identify additional candidates, a small number of which will also be selected for functional study and further follow-up.a) Functional analysis of the Pho1 family of phosphorus transporters/sensors in maize and sorghumThe plant Pho1 genes encode a key family of phosphorus transporters/sensors that have been shown to play a role in the distribution of phosphorus within the plant and in signaling events impacting phosphorus homeostasis. Significantly, partial loss of function alleles of Pho1 in Arabidopsis have been linked to increased phosphate use efficiency.We have previously characterized the Pho1 gene families of maize and sorghum (Salazar-Vidal et al., 2016) and initiated a program of targeted mutagenesis using both endogenous transposon resources and CRISPR gene editing. We will advance the phenotypic characterization of this material, in both greenhouse and field experiments, allowing us to define the role of Pho1 genes, and to address the degree of specialization and overlap between family members.Comparative study of maize and sorghum orthologs of Arabidopsis Pho1 provides an opportunity to study functional divergence in the context of plant nutrient signaling.Phenotypic evaluation will extend preliminary observations indicating impacts of the mutations of root-architecture, plant growth, global transcriptomes and patterns of phosphorus/protein/carbohydrate accumulation in the vegetative parts of the plant and the grain. Although we have several endogenous transposon alleles available, CRISPR material will be needed to provide a complete view of the gene family, and to extend the work to sorghum. This material is in development and is projected to be available for characterization by year 3 of this project. In parallel, we will work through conventional crossing to generate the double and higher-order mutant combinations required to obtain a better understanding of the gene family.

Progress 01/01/20 to 09/30/20

Outputs
Target Audience:The target audience includes other academics,researchers, and students in the plant sceinces field. Changes/Problems:Covid protocols have imposed limitations on travel, recruitment and estabishment of large-scale field trials. In light of this, it is anticipated that computational and in silico activities will dominate over the next reporting period. The project will focus more on the use of existing maize resources/data and less on the development of new sorghum resources than might have otherwise been the case. Collaborative work with researchers both atPSU and other institutions will play an important role, resulting in synergy allowing maintenance of productivity and high quality training opportunities despite the difficult working environment. What opportunities for training and professional development has the project provided?The project has incorporated one postdoc, twograduate students and a project technician. The project team has worked collectively towards the accomplishments described above. Building on backgrounds in plant molecular biology and agronomy, project members have been introduced to maize field genetics and received training in aspects of field design, plant genotyping and the performance/mangement of controlled pollination. Additional training has been provided in computational analysis, including processing and analysis of trial data and quantitative genetic analysis. Project team members have also contributed to manuscript and proposal preparation. Results were also used in undergraduate and graduate courses to teach and train students on cutting edge developments and problems in agriculture/plant science andscientific communication andpresentations. How have the results been disseminated to communities of interest?Results have been presented in manuscripts (both published and as pre-prints), invited research talks, and courses taught at Penn State (AGRO410W: Physiology of Crop Plants andPLANTBIO512: Plant Biology Challenge Problems). What do you plan to do during the next reporting period to accomplish the goals?Goal 1. a) Datasets collected in year one will be used to run predictive models. Material to validate/inform development ofmodels will be advanced in the field. b) The root system architecture/anatomy was characterized in a sorghum line carrying a mutation linked to enhanced Striga resistance in natural populations in West Africa. Further characterization of this material and additional Striga resistant maize material will characterize the relationship between Striga resistance, mycorrhizal mutalism and root architecture/anatomy. c) Data generated in year 1 will be analyzed. Additional replicates/experimental follow-up will be conducted. Goal 2. a) Progress is anticipated in year 2 based on advances in Goal 1. b) The overlap between "environmental" SNPs and genomic regions identified in root trait genome wide association studies will be explored. Goal 3. a) Greenhouse based characterization of eight MAGIC founder parents will be completed. Measurements collected from the greenhouse study will beused to parameterize in silicon modeling root models (Open Simroot) totestfunction under different environmental scenarios. Partially-inbred MAGIC parents will be genotyped. Goal 4. a) A detailed field characterization of maize pho1 mutants will be performed under standard and phosphorus-deficient conditions at the Rock Springs Experimental Farm.

Impacts
What was accomplished under these goals? Goal 1. a) Maize diversity panel root datasets generated by the Lynch group at PSU have been further mined to explore trait correlations and patterns of genotype x environment interaction. Preliminary analyses have been conducted to investigate evidence of selection on root system architecture and anatomy among maize breeding lines. Genome wide marker sets have been used to run a first set of genome wide prediction models. These models will be explored further in the second year of the project. A panel of ~20 parents has been selected to capture aspects of the previously descriptive "steep, cheap and deep" root ideotype for enhanced nutrient capture and water use efficiency. In the 2020 summer field "steep, cheap and deep" parents from the stiff-stalk heterotic groups and a number Protection expired (ex-PVP) stiff-stalk lines were inter-mated to initiate generation of a "root synthetic" for the purposes of future testing predictive models and as a step towards root ideotype selection. b)The root system architecture/anatomy was characterized in a sorghum line carrying a mutation linked to enhanced Striga resistance in natural populations in West Africa. c)A sorghum diversity panel was evaluated with/without inoculation with mycorrhizal fungi in a replicated greenhouse trial. Ionomic analysis of leaf samples is pending. These data will allow characterization of the genetic architecture of the response to arbuscular mycorrhizal fungi in sorghum. Analysis of maize field data was used as the basis of the submitted manuscript "The genetic architecture of host response reveals the importance of arbuscular mycorrhizae to maize cultivation" (see products). Goal 2. a) Progress is anticipated in year 2 based on advances in Goal 1. b) Data linking source and environmental variables has been obtained for panels of maize and sorghum diversity, along with sets of "environmentally-selected" single nucleotide polymorphisms(SNPs). Goal 3. a) Progress has been made in the generation of landrace mapping populations and introgression lines. Partially-inbred families derived from an eight-parent landrace MAGIC (multi parent advanced generation intercross) population will be available for the second year of the project. Greenhouse based characterization of the eight founder parents has been initiated. Goal 4. a) A maize pho1 mutant has been characterized by transcriptome analysis, providing the basis for a paper in preparation. Results confirm the important role for Pho1 and the broader Spexin Hormone(SPX) protein family in the integration of maize phosphorus and nitrogen nutrition. Propagation of additional mutants and generation of double mutants, etc., was carried out in the summer genetics field. Resulting material will be used for field evaluation in the next year of the project.

Publications

  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Erik Diaz-Valenzuela, Ruairidh H Sawers, Angelica Cibrian-Jaramillo, Cis- and Trans-Regulatory Variations in the Domestication of the Chili Pepper Fruit, Molecular Biology and Evolution, Volume 37, Issue 6, June 2020, Pages 15931603, https://doi.org/10.1093/molbev/msaa027
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Alonso-Nieves AL, N Degrees Nez-Rios T, Massange-Sanchez JA, et al. Identification of the maize Mediator CDK8 module, and transposon-mediated mutagenesis of ZmMed12a. The International Journal of Developmental Biology. 2020 Aug. DOI: 10.1387/ijdb.200098sg.
  • Type: Journal Articles Status: Under Review Year Published: 2020 Citation: M. Rosario Ramirez-Flores, Sergio Perez-Limon, Meng Li, Benjamin Barrales-Gamez, Victor Olalde-Portugal, Ruairidh J. H. Sawers. (2020) The genetic architecture of host response reveals the importance of arbuscular mycorrhizae to maize cultivation. bioRxiv 2020.07.06.190223; doi: https://doi.org/10.1101/2020.07.06.190223
  • Type: Journal Articles Status: Under Review Year Published: 2020 Citation: Taylor Crow, James Ta, Saghi Nojoomi, M. Rocio Aguilar-Rangel, Jorge Vladimir Torres Rodriguez, Daniel Gates, Ruben Rellan-Alvarez, Ruairidh Sawers, Daniel Runcie. (2020) Gene regulatory effects of a large chromosomal inversion in highland maize. bioRxiv 861583; doi: https://doi.org/10.1101/861583
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Sawers, R. Mycorrhizal response in maize. Research presentation - "Zeavolution" online discussion group. April 15, 2020.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Sawers, R. Mycorrhizal response in maize. Research presentation - University of Cambridge, UK (online presentation). Paszkowski/Oldroyd root symbioses joint research group meeting. April 21, 2020.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Sawers, R. Mycorrhizal response in maize. Research presentation - North Carolina State University (online presentation). Plant biology seminar. July 24, 2020
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Sawers, R. Introduction to Quantitative Genetics. Workshop in Quantitative Genetics. Langebio-Cinvestav, Mexico (online presentation). Interaction with graduate students in Integrative Biology program. August 31, 2020 - September 4, 2020.