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ARS Home Pacific West Area Aberdeen, Idaho Small Grains and Potato Germplasm Research Research





- Esvelt Klos, Kathy
- Gao, Dongying
- Hu, Gongshe
- Yimer, Belayneh
- 2023
- Publications
- Plant Genetic Resources, Genomics and Genetic Improvement (301)

Research Project: Enhancing Barley and Oat Productivity, Quality, and Stress Resistance **Location:
                            Small Grains and Potato Germplasm Research** 2023 Annual Report **Objectives**
                                

Objective 1: Exploit crop genetic resources to identify and characterize genetic variation contributing to barley and oat quality, disease resistance, and stability in the face of environmental variation. 
Sub-objective 1.A: Screen and map stripe rust resistance in wild barley accessions.
Sub-objective 1.B: Characterize Fusarium head blight (FHB) resistance in barley and develop disease resistant germplasm.
Sub-objective 1.C: Map oat genes conferring resistance to crown rust disease and develop resistant germplasm. 
Sub-objective 1.D: Identify and map the locations of genes contributing to oat response to environmental stress. 

Objective 2: Use gene editing technology to change barley candidate gene expression and determine the impact on seed composition (seed storage proteins, enzymes). 

Objective 3: Develop barley and oat germplasm, adapted to irrigated, dryland or organic cropping systems, with increased yield, better quality, and superior resistance to stress. 
Sub-objective 3.A: Develop No-Glycosidic Nitrile (No-GN) barley lines suitable for brewing and distilling. 
Sub-objective 3.B: Develop further improved winter food barley varieties. 
Sub-objective 3.C: Develop organic barley and oat lines. **Approach**
                                

Identification of desirable traits within wild or un-adapted barley and oat accessions and introgression into elite germplasm can create a basis for developing new cultivars (Objective 1). Genetic data for wild barley accessions from the National Small Crains Collection will be used to establish a molecular ID database (Sub-objective 1A). Association mapping methods will be used to identify QTL for Barley Stripe Rust resistance (Sub-objective 1A) and for oat seedling growth (emergence, growth per day, tillering) under moisture stress (Sub-objective 1D). Linkage analysis will be used to map barley resistance to Fusarium Head Blight (Sub-objective 1B) and to validate QTL contributing to oat grain quality (test weight, GC, kernel size distribution), and phenotypic stability (Sub-objective 1D). Both association and linkage methods will be used to map Oat Crown Rust resistance QTL (Sub-objective 1C), with digital imaging used to capture measures of disease progression. Superior selections may be evaluated for release as new varieties and will be used as parents in new crosses.
Under the hypothesis that editing seed storage protein transcription factors will reduce barley seed protein content (Objective 2), Hvu_NAC genes will be CRISPR edited using guide RNAs targeting exons. The transgenic barley plants and the wild types will be evaluated for seed protein and starch content. Although it is possible that editing Hvu_NAC genes will not significantly reduce the seed protein content, this would also be informative of the genetic basis of barley seed protein regulation.

This project will develop barley and oat germplasm, adapted to irrigated, dryland or organic cropping systems, with increased yield, better quality, and superior resistance to stress (Objective 3). Agronomic performance will be assessed on key traits such as yield, lodging resistance, drought tolerance, grain test weight, grain plumpness, and maturity. Elite lines will be tested for at least three years in replicated, small plot yield trials at multiple Idaho locations. Low/no glycosidic nitrile cultivars or elite lines for brewers and distillers (Sub-objective 3A) will be developed using marker-assisted selection of the No-GN allele from Full Pint. Selected lines will be evaluated for malting quality based on standards of the American Malting Barley Association. Food barley with a fall-planted growth habit and good winter survival will be improved for higher ß-glucan content and yield (Sub-objective 3B). Lines grown as fall-planted headrows that survive multiple winter tests will be considered winter types. Those with consistently good agronomic performance, and high ß-glucan content will be selected for entry into advanced trials. Barley and oat elite lines will be evaluated for performance under organic production (Sub-objective 3C). Twenty barley and 20 oat lines will be grown under three management regimes (conventional irrigated, conventional water stressed and organic irrigated) over four years to determine the lines best suited to organic production in the Intermountain West; and whether evaluation in an organic nursery provides unique information about line performance. **Progress Report**
                                

This project began in February 2023 continues research from expired project 2050-21000-034-000D. For additional information, see the report for the expired project. 

The goals of this project are to produce information that will increase the breeding efficiency of barley and oat, and to develop improved barley and oat germplasm and varieties that meet the needs of expanding and established market classes in the face of complex disease and environmental pressure.

Progress was made toward increasing breeder and grower options for robust and resilient resistance to barley stripe rust disease (Sub-objective 1A). Barley germplasm carrying a combination of seedling stage and adult plant stage resistance donated by both an unadapted heirloom variety and an elite Aberdeen malting line were evaluated for disease resistance and agronomic performance in multi-year, multi-location tests. Disease resistance in the field was found to increase with each resistance gene region incorporated, while agronomic performance was maximized in lines carrying three resistance alleles from the heirloom variety in combination with the maximum proportion of the genome inherited from the elite parent at all other locations. This information has been submitted for publication to the journal PytoFrontiers. Germplasm will be available to breeders upon request. Towards identifying new sources of resistance, 1,906 accessions of wild barley (Hordeum spontaneum) were obtained from the National Small Grains Collection. Seed of these were planted along with 10 cultivated barley check varieties to obtain sufficient seed for future characterization. We successfully obtained viable seed and tissue for use in genotyping from 1,900 accessions.

Initial evaluation of spring malting barley bi-parental populations for reaction to Fusarium head blight (FHB) disease and accumulation of Deoxynivanol (DON) mycotoxin was performed for all four crosses made with two elite Aberdeen breeding lines by two North Dakota varieties to investigate the character and uniqueness of Aberdeen, Idaho, FHB resistance (Sub-objective 1B). These populations were planted in 2023 FHB evaluation nurseries at one location in Idaho and two locations in North Dakota to obtain multi-year data that will meet a high publication standard. Preliminary evaluation suggests that there may be lines within these populations with greater FHB resistance than the parents.

A genetic resource for identifying and mapping genes that contribute adult plant resistance to oat crown rust disease has been completed and partially genotyped (Sub-objective 1C). This nested association-mapping population shares a high-yielding oat parent from the Aberdeen breeding program (HA08-03X21-1) combined with one of six heirloom varieties chosen from the National Small Grains Collection for resistance to oat crown rust disease. 2023 summer seed increases will provide the seed needed for field evaluations of crown rust resistance. Analysis of this full genetic resource will allow us to map the locations of resistance genes, compare them for the size of their effects on disease resistance and determine if each resistance source has the potential to contribute unique genetic resistance to new varieties.

Towards Sub-objective 1D, five planned oat crosses (HA05AB29-39 X HA08-03X09-1; HA05AB29-39 X ND060235; HA05AB29-39 X IL2294-1; HA08-03X09-1 X Bountiful; HA08-03X09-1 X H927-1-6-X-X_24) have been made and populations are under advancement by the single-seed descent method. Once complete, these populations are expected to provide information that will validate the location and effect of several milling quality quantitative trait loci (QTL) that were previously identified using whole-genome association mapping. These include QTL that are anticipated to decrease the variability in milling quality of oat produced in different production environments without reducing quality under optimal environments.

To investigate the genetic control of barley grain protein content (Objective 2) two transcriptional factor gene sequences, Hvu_NAC1 and Hvu_NAC2, were chosen as candidates for experimental investigation. These genes will be targeted for CRISPR-edited deletions. Towards this, guide RNAs (short RNA sequences specific to the candidate genes), and the associated transformation vectors were designed and constructed. The accuracy of the vector insert fragments for both plasmids were verified by sequencing. Once transformed into barley plants, the CRISPR-associated endonuclease is expected to cut the double-stranded DNA of our candidate genes to produce a targeted mutation.

Elite germplasm for malting and food markets was evaluated and presented to stakeholders (Objective 3). One spring (16ARS067-13) and two winter (11ARS652-7 and 13ARS526-8) malting barley breeding lines were submitted for pilot scale testing to the American Malting Barley Association. Spring food barley elite lines have been produced to target yield improvements. Also under Objective 3, an elite oat breeding line (HA08-03X21-1) has been identified for variety release. Based on multi-year evaluations, this line has significantly higher yield potential than current varieties under conventional production.

Barley germplasm without Glycosidic Nitrile (No-GN) is suitable for both malting and distilling (Sub-objective 3A). For this purpose, 20 genetically stable lines have been produced that carry the molecular genetic markers for the No-GN trait, donated by the cultivar ‘Full-Pint’, which were selected from a total of 107 lines based on their field performance. These will undergo chemical characterization of their GN phenotype and be entered into more advanced agronomic and malting quality evaluations. Additional crosses will be used to expand the presence of this trait in the Aberdeen, Idaho, malting barley breeding program. These activities have been presented to stakeholders.

There is a great deal of room for improvement to winter food barley germplasm (Sub-objective 3B). Following the release of the 'Upspring’ variety, focus was shifted to yield, lodging, and winter survival. Elite breeding lines 05ARS849-15, 12ARS777-1, 14ARS733-4, 14ARS744-10, 14ARS744-16, 14ARS766-5, 16ARS844-1 and 16ARS846-7 are in agronomic and quality evaluations.

To validate the performance of the new Aberdeen permanent organic small trains nursery while simultaneously evaluating elite barley and oat for suitability to organic production, an experimental design has been completed (Sub-objective 3C). Twenty barley lines and 20 oat lines have been selected to include the most promising malting barley, food barley and oat breeding lines. Replicated multi-year testing in both organic and conventionally managed evaluation plots will provide a proof-of-concept for use of the organic nursery in variety development. **Accomplishments**


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