CIHEAM-Zaragoza and University of Lleida (Spain); Ignacio Romagoza, Ramzi Belkodja, Ana Casas, Alba Farré-Martínez, Ernesto Igartua, and Jose Miguel Soriano
-
The OWB has been used for more than a decade in an international program run by CIHEAM-Zaragoza and the University of Lleida (https://www.iamz.ciheam.org/agendas/xxiii-international-master-in-plant-genetics-genomics-and-breeding/ ). In this two-year biennial program, 20 to 25 diverse students, from around 10-12 countries, mostly from West-Asia, North-Africa and Europe, participate. For the sake of simplicity we use a reduced set of 82 DHL of the H.b. set and 252 markers. Students are responsible for the phenotypic characterization of field grown OWB double haploids, segregating for the following qualitative and quantitative traits: Plant Height; Length of the last internode; Spike length; Awn length; 2 vs 6 Rows; Compact vs. loose spike; Hooded vs Normal spikes; Phenological Cycle. Based on their own phenotypic and previously available genotypic data, a quantitative genetic analysis are conducted using the freely-available public software QTL IciMapping 4.2 for both map construction and QTL mapping.
Cornell University (USA); Susan McCouch
-
At Cornell University, the OWB population consisting of 94 H.b.-derived DH lines was every year for over 20 years as part of an introductory course entitled “Plants, Genes and Global Food Production”. The population and the two parents were planted in a greenhouse in Ithaca, NY in mid-August, at the beginning of fall semester, and plants were fully mature and ready for a hands-on learning lab by the first week in November. Students worked in groups of 3 to collect phenotypic data on leaf- and spike-morphology traits for the entire population, guided by photographs of the mutations observed in the parental lines. Each group of students was assigned a different set of 3 mutant traits, with overlapping trait observations across groups. The class was able to evaluate 6 mutant phenotypes: two- versus six-rows (vrs1), long versus short awns (lks2), hooded (no awn) versus normal floret (awn) (kap), dense versus normal head (zeo), leaf pubescence versus no pubescence (pub), and hairs versus no hairs on the lower leaf sheath (hsh). As part of an in-class exercise following the lab, phenotypes were cross-checked among groups and compared to the photo gallery of DH line spike morphologies (available on the OWB web-site) to resolve any differences. Datasets were then merged to generate a common dataset for the population as a whole as the basis for an exercise on linkage mapping. Linkage was analyzed using the phenotypic information generated by the students in combination with the genotypic information available on the web-site with chi-square tests to test for linkage. The OWB population captured student interest and heightened their awareness of the relationship between plant genetics and plant breeding. It clarified the relationship between genes and alleles, deepened understanding of the laws of Mendelian inheritance and segregation and provided an opportunity to discuss how linkage between genes and molecular markers is used to predict phenotypic outcomes in both human genetics and crop improvement.
Huntington University- Indiana (USA); Raymie Porter
-
The ISS is used in an upper-level undergraduate plant breeding course to train students in recognizing different traits and phenotypes. Students had little or no prior experience in recognizing traits of small grains, and were given only basic guidance in how to discover and recognize phenotypes. The objective of these exercises was to develop the skill of phenotype recognition through careful visual observations. Initially (while plants were in the early stages), students began observing and noting phenotypic differences between the two parental lines. Next, they looked for visible or measurable differences in the 18 non-parental lines based on the differences between the parental lines. As the plants developed, they continued their observations and added more qualitative and quantitative traits to their list. They measured traits that were quantitative (i.e., height, number of tillers), and devised a relative (1-5) rating scale for other traits that were more difficult to measure accurately (awn length, tiller erectness). They also categorized those traits that were qualitative (stem color, pubescence, spike color, awned vs hooded, number of rows, etc.). In their final report for the exercise, they each returned a spreadsheet with their individual plant observations for each trait, the mean, maximum, and minimum for each quantitative trait, and the number of lines of each qualitative trait phenotype. Future iterations of the course may include a basic linkage mapping exercise using published data from the full OWB set.
Iowa State University-USDA/ARS (USA); Roger Wise, Gregory Fuerst, Nick Peters, Nancy Boury, Laurie McGhee, Melissa Greene, Sarah Michaelson, Julie Gonzalez, Nick Hayes, Ron Schuck, Lance Maffin, Garrett Hall, Taylor Hubbard, and Ehren Whigham
-
iTAG (Inheritance of Traits and Genes) uses morphogenetically diverse Oregon Wolfe Barley in laboratory and classroom activities to connect visible traits to identifiable differences in DNA sequences. iTAG focuses on three traits to illustrate basic concepts in plant development, domestication, and disease resistance. Participants learn DNA extraction, PCR, gel electrophoresis, and document DNA polymorphisms among different phenotypes, concepts critical to producing productive crops. iTAG barley has been used by nearly 50 instructors in >200 high school and community college biology classes from 2010-2023, impacting nearly 5000 students, of which one third were from underrepresented groups from urban to rural communities.
Norweigan University of Life Sciences (Norway); Anja Karine Ruud, Morten Lillemo, Øyvind Jørgensen
-
Out of five practical exercises in Plant Breeding (Bio 248), the population is used in three:
-
Qualitative traits – the students phenotype grain and plants for different qualitative traits (Vrs1/vrs1, Nud/nud, Zeo/zeo, Lks2/lks2, Kap/kap). The interaction between Lks2 and Kap is used to understand epistasis.
-
Quantitative traits – the students measure plant height, and use the Zeo-results from the previous exercise to discuss genes with major and minor effects on plant height.
-
QTL mapping – a computer exercise in R, were they do QTL mapping of qualitative and quantitative traits from the OWB population. Since the computer skills vary, they can use phenotype data we provide, which is formatted for easy import into R, or whether they want to try with their own data from the two previous exercises. Focus is on interpreting the results – and it is helpful that the students have seen and measured the traits themselves.
Oregon State University (USA); Pat Hayes and Kelly Vining
-
The Oregon Wolfe Barley population is used at OSU in undergraduate and graduate-level courses in the Plant Breeding and Genetics (PBG) program (https://catalog.oregonstate.edu/courses/pbg/.
-
In the undergraduate/introductory graduate course (PBG430/530), the OWBs are used to demonstrate the principles of Mendelian analysis and its continued relevance in an era of abundant and cheap whole genome sequence.
-
In the graduate course (PBG620,621), the OWBs are used to demonstrate molecular marker development and application. The course has two parts: Introduction to Molecular Markers, and Genetic Mapping and Association. A hands-on lab exercise in the second part engages students in phenotyping, measuring spike length and categorizing awned vs. hooded floral morphology. This allows students to work directly with plant material instead of simply downloading online datasets, although the online option is available for students attending remotely. The phenotype data is then used in genetic mapping and genotype-phenotype association exercises. Student assessments consist of a lab report, an in-class quiz, and a comprehensive final exam covering all material from the term, including the OWB exercises.
-
Mendelian inheritance
-
Qualitative and quantitative inheritance
Purdue University (USA); Diane Wang, T.C Ting, M.M. Hammons, and C. Hoagland
-
The ISS was recently grown at Purdue University’s controlled environment phenotyping facility, which employs an automated system with pots on conveyer belts to collect plant measurements using imaging (https://ag.purdue.edu/aapf/virtual-tour.html). For the ISS evaluation, the facility’s RGB camera system was leveraged to acquire color images of each plant (top and 12 side views), automatically segment the images, and extract predefined parameters to quantify growth and color related traits of each line three times per week. Alongside these image-based traits, ground reference observations of plant height and tiller number were collected two times per week over the course of the 42-day evaluation period. This gives rise to a 324-observation dataset containing both image-based metrics and ground-reference data, which may be used for a variety of downstream instructional applications. These may include (1) teaching descriptive statistics and exploratory plotting of image-based and manual measurements and (2) teaching simple modeling of ground reference measurements using various combinations of image-based variables. These kinds of exercises may be most appropriate for high school or university-level students. All data along with experimental details are publicly available at Zenodo 10.5281/zenodo.10999418
Universidad Politécnica de Madrid (Spain); Elena Benavente, Estela Giménez, Patricia Giraldo, Carla Guijarro Real, Marina Martínez García, and Laura Pascual Bañuls
-
Faculty from our program (https://blogs.upm.es/geneticaymejora/en/) use the DH H.b. Oregon Wolfe Barley collection in various courses related to plant breeding at both undergraduate and master's levels, with adjustments according to the academic level. Each academic year, the DH collection is grown in triplicate under greenhouse conditions, where students measure plant height and spike length. Additionally, during laboratory sessions, students genotype the collection using two PCR-based molecular markers: the Knox-dup dominant marker (a duplication) and the Bmac 0130 co-dominant microsatellite (SSR) marker. Using phenotypic and molecular data, along with a molecular marker matrix provided by the faculty, students conduct QTL mapping associated with the quantitative traits using the free software MapDisto (http://mapdisto.free.fr/). This practice allows students to understand the importance of accurate phenotyping and genotyping, the differences in the analysis of dominant and co-dominant molecular markers, and how to perform association studies between molecular markers and complex traits.
-
In addition, we have developed and published resources for using an F2 population derived from the OWB-D x OWB-R.
University of Bologna (Italy); Silvio Salvi and Giuseppe Sangiorgi
-
The 93 OWB A.C._93 lines are used in the practical activities of our course (https://corsi.unibo.it/2cycle/PlantAgriculturalBiotechnology). Objectives of the activities are to expose the students to Mendelian (qualitative) and quantitative types of genetic variation and provide hands-on experience in phenotype collection, biometrical analysis and genetic mapping of both Mendelian loci and QTL. Each OWB_A.C. _93 DH line is grown in pots in a replicated design (4 plants per line) in greenhouse. Mendelian traits scored are: hooded (presence/absence), awn (presence/absence), row type (two/six). Quantitative traits measured are: awn length, spike length, number of florets, plant height. We take advantage of publicly available marker data. QTL mapping is run using MapQTL6 (Ooijen V. 2009). Quantitative trait data are used for running standard biometrical analysis. Trait correlation analysis is used to show statistically significant correlation between plant height with spike length and not with number of florets. QTL mapping by single marker and interval mapping analysis for both plant height and spike length show a strong LOD peak on chr. 2H, at approximate cM 150-160, in correspondence of ZEO-1 locus, illustrating its pleiotropic effect. Results are used to illustrate the concept of QTL genetic effect, proportion of explained phenotypic variation and the indefinite separation between QTL and Mendelian loci. In the next academic year, variation for root architectural traits (number of seminal roots, root growth angle, total root length, etc) within OWB_A.C. _93 at the seedling stage will be addressed using a semi-hydroponic protocol, followed by QTL analysis.
University of California - Davis (USA); Pat Brown
-
"The cool thing is that you can see your Manhattan plot change in real time as you enter phenotype data. I’m still amazed that we can get such nice mapping results with just 25 genotypes, but I guess that’s the result of a) Mendelian traits; and b) good genotype data on completely homozygous lines."
University of Dusseldorf (Germany); Rebecca Schuller, Maria von Korff, and Agatha Walla
-
We integrate the OWB DH population into the second-year bachelor’s curriculum of the interdisciplinary program “Quantitative Biology” (https://www.qbio.hhu.de/en/) within the Cluster of Excellence on Plant Sciences (CEPLAS), which has up to 40 enrolled students annually. Our lecture series covers critical population and quantitative genetics topics, laying the theoretical groundwork for the following practical sessions. Students engage hands-on with the OWB DH lines to gain experience in phenotypic data collection in the greenhouse. Divided into groups, they measure spike length and plant height as quantitative traits. Each group is also assigned two qualitative traits, including row-type, short or long awns, hooded florets, naked caryopsis, black lemma and pericarp, and leaf variegation. Students can refer to images on https://barleyworld.org when uncertain about specific traits such as caryopsis color or hull adherence. Using the R software package R/qtl (Broman et al., 2003) and marker and linkage data from https://barleyworld.org, students are introduced to the fundamentals of linkage mapping and QTL analysis. They begin by constructing their own barley linkage maps using a selection of markers. Next, using the entire marker and linkage data, the students investigate the linkage of their qualitative phenotypic traits with markers. Measurements for plant height and spike length are used to perform QTL analysis in R. Finally, the students compile their results into a publication format, simulating the identification of genes for the investigated traits and reinforcing their understanding of the concepts.
University of Illinois at Urbana-Champaign (USA); Juan Arbelaez
-
At the University of Illinois, first-year students in Introduction to Crop Sciences (CPSC 112) grow the entire H.b. OWB population and collect data on spike morphology traits such as two- versus six-rows, the presence or absence of awns, and their length, among others. This data substantiates the concepts of genes, alleles, and Mendelian Law of Segregation. We extend the data by adding different molecular markers, linked and un-linked to these traits. Students create pairwise genotypic classes from trait-marker combinations count and analyze them using chi-square tests to illustrate the concepts of parental and recombinant genotypic classes, the effect of linkage on recombination, and how scientists implement these analyses to identify the location of genes within the genome (see a link to UIUC lab material). The OWB population not only provides an empiric and interactive experience of plant genetics, but it is also a first exposure to how genetics can be used to improve our crops.
University of Minnesota (USA); Katy Guthrie and Robert Stupar
-
Beginning in 2012, the University of Minnesota developed a semester long exploration of the H.b. subset of the OWB population for an upper-level, Plant Genetic Resources course. The project spans eight labs, including one in-lab writing workday, over a 14-week semester. Students take on the role of a researcher exploring connections between phenotype and genotype using tools and techniques commonly used by molecular geneticists and breeders. Learning is scaffolded; students are first assigned a unique set of individuals they are responsible for characterizing. This includes collecting phenotypic data on growing plants described in lab one, and digital images described in lab five, as well as genotypic data as outlined in labs two through four. This includes running an electrophoresis-based Bkn-3 PCR marker (Muller et al 1995) to predict the hooded phenotype while the plants are still in the seedling stage (Figure 2). Individual data is placed in a combined class document and interpreted in context of the larger dataset. Class data, combined with publicly available data in cases of small class size, is used to complete labs six (genetic linkage) and seven (gene mapping). Starting with activities that promote individual learning buildings students’ confidence in core concepts before engaging in group learning of more difficult concepts, like epistasis and linkage. Both formative and summative assessments are used to gauge student learning. Formative assessment includes keeping a lab notebook for participation points. Summative assessments include low stakes “Learning Readiness Quizzes” to prepare students for lab activities, and a comprehensive OWB Lab report where students summarize their learning throughout the semester.
-
Materials, including lab manuals, worksheets, activity workbooks, and assessments (including rubrics), are available this Google Drive Folder.
Archives
-
Oregon State University - Linkage mapping and dissection of the hooded phenotype (Kelly Vining and Pat Hayes) 11/2020
-
Oregon State University - Monohybrid segregation and hypothesis testing using Vrs1 sequence SNPs (Alfonso Cuesta-Marcos and Pat Hayes)
-
IAMZ - linkage mapping and QTL analysis (Alba Farre)