USDA-ARS Barley Research at Aberdeen, Idaho

United States Department of Agriculture (USDA)Agricultural Research Service (ARS)

Small Grains and Potato Germplasm Research UnitAberdeen, Idaho

Unit Snapshot

• 40 permanent employees– 12 scientists– 29 support staff

• 5 post-doctoral fellows (2 in barley)• 3 locations

– Aberdeen, ID (9 scientists)– Hagerman, ID (2 scientists)– Bozeman, MT (1 scientist)

University of Idaho Research and

Extension Center, Aberdeen, ID

University of Idaho Fish Culture Research

Station, Hagerman

National Small Grains Germplasm Research Facility and

University of Idaho Research and Extension Center

within the Aberdeen Greater Metro Region

USDA-ARS National Small Grains Germplasm Research Facility located at the

U. of Idaho Aberdeen Research & Extension Center

Five Projects in the Aberdeen Unit

– National Small Grains Collection--Harold Bockelman, Mike Bonman

– Improving Trout Production--Rick Barrows, Ken Overturf, Keshun Liu, Tom Welker

– Plant and Seed Chemistry Genetics--Victor Raboy

– Genetic Improvement of Barley and Oats for Enhanced Quality and Biotic Stress Resistance-- Phil Bregitzer, Mike Bonman, Gongshe Hu, Kathy Klos

– Potato Improvement--Rich Novy, Jonathan Whitworth

Hordeum Accessions by Country of Origin

• Total of 98 countries• Largest countries:

United States 8720Ethiopia 4216China 2182Turkey 1994Israel 1161

Kernel diversityin Ethiopianbarley landraces

The USDA Barley Core Collection: Genetic Diversity,Population Structure, and Potential for Genome-WideAssociation Studies. 2014. Munoz-Amatriain et al.

Barley, 12% protein

Pretreatment, low temperature

Barley Protein Conc., ~41% protein

Barley Protein Conc., ~55-60% protein

Ethanol

Low temperature treatment, fermentation, no distillation

Barley Protein ConcentrateARS and Montana Microbial Products, LLC

Alternative Ingredients for Aquafeeds

Fishmeal cost/MT: ~$600 in 2004,~$2100 in 2014

Single Ingredient Replacement for Fish mealBarley Protein Concentrate or Bio-soy

Gai

n, g

ram

s/fis

h

bc LM

ab

a

M

N

c

L

Fish meal BPC or Bio-soy

100% 33/66 66/33 100%

Barley Protein Concentrate

Pilot Plant in operation in Wisdom MT, supported by an Idaho aquaculture feed producer.

Pilot Plant is producing ~50 tons per month of BPC for full fish farm testing.

Product being sent to major feed company in Norway for further testing.

Full scale production plant will produce 500 tons/ month of BPC, with a 20% yield, so 2500 tons of barley needed per month.

Epigenetics in Barley: Trans-Generational Adaptation to Macronutrient StressParental Exposure to Macronutrient Stress Greatly Enhances Progeny Growth and Yield

When Grown Under a Similar StressPanel A: Parental Generation, cv. Harrington: the plant on the left was grown on optimal nutrition (10 mM N, 1.5 mM P, 4 mM K, illustrated as “NPK”); the plant on the right was grown on severely limiting nutrition (0.1 mM N, 0.06 mM P, 0.06 mM K, illustrated as “npk”). Seeds were harvested from five replicates, bulked and used to plant next generation illustrated in Panel B. Panel B: Pots 1 through 4 were planted with seed harvested from parents grown with optimal macronutrient nutrition (Panel A, left), and Pots 5 through 8 were planted with seed harvested from parents grown with severely reduced macronutrient nutrition (Panel A right). All eight pots were watered with the same nutrient solution, representing a moderately reduced macronutrient nutrition ( (3.0 mM N, 0.3 mM P, 0.8 mM K). Measures of plant growth and grain traits for plants in Panel B are given in Table 1.

1 2 3 4 5 6 7 8Pot Number

A B

NPK npk

Table 1. For plants illustrated in Panel B, measures of plant growth, grain yield and leaf and grain phosphorus (P) were obtained. The data represents the mean (±standard deviation) of the four replicates illustrated in Fig 1B.

ParentalNutrition

Progeny Traits When Grown on Moderate Macronutrient StressPlant

HeightNo. of Tillers Leaf P

Grain Weight GrainYield Grain P

cm mg g-1 mg grain-1 g plant-1 mg g-1 mg plant-1

Optimal 13±0.8 13±6 3.3±0.6 33±6.5 6.0±4.3 5.4±0.8 30.1±17Reduced 19±2.4 26±11 3.1±0.2 48±7.7 15.9±3.8 4.6±0.7 73.5±17

Results from observing Lenetah X GZ progeny

• The seedling resistance gene from GZ is on chromosome 4H.• There may also be genes influencing resistance on 6H and 7H.

In progeny from 95SR316A X GZ:1. Measure molecular markers on both ends of

the gene region and also in the middle.2. Did the most resistant progeny get that part

of chromosome 4H from GZ?

95SR316A X GZ

R MR MS S

68 lines w/GZ @ 4H

13 42 8 5

165 lines total 16 58 38 53

Three potential targets for marker-assisted introgression of malting quality QTL from ‘Stellar’ into the Aberdeen breeding program

Chr Location (interval) Trait(s) LOD Genotype at the target region of lines meeting AMBA malting targets

5H 173.5 (164.1‐182.4) cM DP 2.93‐3.32

6H 57.2 (55.9‐57.5) cM S/T 19.63

6H 70.7 (66.5‐89.7) cM(May be >1 QTL)

DP (wort protein, ‐

glucan)

2.74‐7.38

01Ab8219 X‐over Stellar

4 3 11

01Ab8219 X‐over Stellar

9 2 14

01Ab8219 X‐over Stellar

3 5 17

Three potential targets for marker-assisted introgression of malting quality QTL from ‘Stellar’ into the Aberdeen breeding program

Chr Location (interval) Trait(s) LOD Genotype at the target region of lines meeting AMBA malting targets

5H 173.5 (164.1‐182.4) cM DP 2.93‐3.32

6H 57.2 (55.9‐57.5) cM

S/T 19.63

6H 70.7 (66.5‐89.7) cM(May be >1 QTL)

DP (wort protein, ‐

glucan)

2.74‐7.38

01Ab8219 X‐over Stellar

4 3 11

01Ab8219 X‐over Stellar

9 2 14

01Ab8219 X‐over Stellar

3 5 17

S/T QTL Region

DP QTL Region

Rare alleleCommon allele

Malt QTLs– next steps

• Our primary target for marker-assisted introgression will be the S/T QTL at 57 cM on Chr. 6H (LOD=19.63)

• Haplotype diversity in this region of 6H is relatively low in the Aberdeen germplasm

• We will develop populations segregating for the ‘Stellar’ S/T QTL along with various combinations of chromosome 6H haplotypes already present in the Aberdeen breeding material

1. Fusarium graminearum strain TRI5 prom::GFP  expressing Green Fluorescent Protein marking mycotoxin production and Red Fluorescent Protein marking fungal growth (Ilgen et al., 2008)

2. In vitro exposure of  21mer siRNA targeting upstream transcription factors TRI6 & TRI10 that regulate  production of DON via TRI5 induction

3. Fluorescence is measured by plate reader as fungus grows in DON‐inducing conditions

Goal: Transgenic barley plants resistant to Fusarium head blightApproach: Test construct components directly in Fusarium (fast!)Project: Test effectiveness of siRNAs for host induced gene silencing

TRI10 TRI6 TRI5 DON

Trichothecene Pathway

TRI5prom::GFP GFP

siRNATranscription factors

Trichothecene Synthase

Fluorescent plate reader: Biotek Synergy H1

Output data

Biomarker for DON

Results for top siRNA (targets TRI6)Exposure of certain siRNAs decreased Green Fluorescent Protein expression, indicating low induction of TRI5 and therefore lower DON production. No significant effect on fungal growth was observed.

Barley Breeding Activity

• Covers both malting and food types• 50-50 winter/spring; predominantly 2-row

malting types• Yearly 8-10 K evaluation plots and 5-7 K

head rows• Six winter and nine spring locations, plus

NZ off-season nursery

Promising Spring Malt Lines

• 2Ab04-X01084-27 in AMBA plant scale testing

• 2Ab07-X031098-31 is available for AMBA plant scale testing– Yield potential equal to that of Voyager– Improved lodging resistance, higher extract,

lower beta-glucan

Multi-year Data for 2Ab07-X031098-31

LD = Lodging

• 02Ab669• 100 acres sown for AMBA plant scale testing• Better yield, lower beta-glucan, and lower

protein than Charles

Promising Winter Malt Lines

Entry Yield(Bu/A)

Winter Survival(%)

Extract(%)

Protein (%)

DP BG(ppm)

FAN(ppm)

05ARS540-125 143.4 87.8 81.5 11.4 127 144 18602Ab669 153.6 73.4 81.9 10.3 95 98 179Charles 141.4 64.4 82.4 11.1 108 140 207Endeavor 144.2 83.6 81.2 11.0 119 129 191

High beta-glucan Food Barley

• Cultivar in commercial use: Transit ~10% beta-glucan

• New release: 2Ab09-X06F058HL-31, beta-glucan = Transit, 10% better in grain yield

• ‘CM1’ an ultra-high beta-glucan mutation--13-18%

• New generation of food barley lines in the pipeline with yield of 2Ab09-X06F058HL-31 but 30% higher beta-glucan

More Germplasm News from AberdeenAll the barley that’s fit to breed!

Low phytate germplasm: Development concluding with the release of two lines: *‘Harriman’: hulled; higher yield (= Baronesse), test weight, available P vs ‘Herald’*‘Selway’: hulless; higher yield (↑ 8%) and test weight vs Clearwater

Yield drag from LPA has yielded to recurrent selection

Russian wheat aphid resistant barley: many germplasm lines and cultivars releasedvia collaboration with ARS‐Stillwater; in 2014:

*RWA1758 was licensed and is now available to producers*SNP‐based QTL mapping used to categorize resistance sources and germplasm lines

Transposon tagging barley population:*70 lines developed at Aberdeen released into NSGC; documentation in NCBI*62 lines developed at UC Berkeley to be released in 2015*Delivery of transgenes as recombinant Ds transposons will create many more

tagged lines as a by‐product. Development depends on community interest!

Thank you