Barley Genetics as a Model Barley Genetics as a Model for International for International

Collaborative ResearchCollaborative Research

Lauren OsbornLauren OsbornOregon State UniversityOregon State University

Department of Crop and Soil ScienceDepartment of Crop and Soil Science

Mentors: Dr. Patrick M. HayesMentors: Dr. Patrick M. Hayes

Dr. Peter SzucsDr. Peter Szucs

Dr. Tony ChenDr. Tony Chen

ChaptersChapters

Inheritance of Final Leaf Number in Dicktoo x Inheritance of Final Leaf Number in Dicktoo x Calicuchima and Dicktoo x OWB-D FCalicuchima and Dicktoo x OWB-D F22 populations populations

International and Local Research ExperienceInternational and Local Research Experience

Oregon Wolfe Barley Oregon Wolfe Barley

Inheritance of Final Leaf Number in Dicktoo x Calicuchima and Dicktoo

x OWB-D F2 populations

BarleyBarley

Winter typesWinter types Plants survive harsh winter temperaturesPlants survive harsh winter temperatures Vegetative StateVegetative State

Facultative TypesFacultative Types Low temperature tolerant but do not require cold Low temperature tolerant but do not require cold

periodperiod

Spring typesSpring types Planted in the springPlanted in the spring No need to survive winter temperaturesNo need to survive winter temperatures

Vernalization RequirementVernalization Requirement

Induction of flowering by exposure to low temperatureInduction of flowering by exposure to low temperature

No VRN No VRN VRN

Spring

FacultativeWinter

SensitivitySensitivity

Growth Growth HabitHabit

Winter Winter HardinessHardiness

SensitivitySensitivity

VernalizationVernalization

RequirementRequirement

SensitivitySensitivity

Short day Short day PhotoperiodPhotoperiod

SensitivitySensitivity

WinterWinter ++ ++ +/-+/-FacultativeFacultative ++ -- +/-+/-SpringSpring -- -- --

Vernalization Requirement

Photoperiod Sensitivity

Low Temperature

Tolerance

Cereal Winter Hardiness

Low Temperature

TolerancePhotoperiod Sensitivity

VernalizationVernalization

Two genes determine vernalization Two genes determine vernalization requirement in barley requirement in barley (Von Zitzewitz, 2005)(Von Zitzewitz, 2005) VrnH1VrnH1 VrnH2VrnH2

Winter alleles at both loci need to be present Winter alleles at both loci need to be present for the plant to require vernalizationfor the plant to require vernalization

VrnH1VrnH1

VrnH1 is located on chromosome 5HVrnH1 is located on chromosome 5H

Involved in plant growth and developmentInvolved in plant growth and development

ExpressionExpression Plant undergoes a vegetative to reproductive Plant undergoes a vegetative to reproductive

transitiontransition Meristem differentiation and the plant growth Meristem differentiation and the plant growth

cycle cycle

VrnH2VrnH2

VrnH2 is located on chromosome 4HVrnH2 is located on chromosome 4H

VrnH2 encodes a repressorVrnH2 encodes a repressor Effects VrnH1 Effects VrnH1 Prevents its expressionPrevents its expression

Repressor is activeRepressor is active VrnH1 is not expressed VrnH1 is not expressed Plant remains vegetativePlant remains vegetative

Vrn H2Vrn H2

Winter typesWinter types Repressor prevents expression of VrnH1Repressor prevents expression of VrnH1 Until a period of low temperatureUntil a period of low temperature

Cold period achievedCold period achieved VrnH2 genes cease expressionVrnH2 genes cease expression No repressor protein is producedNo repressor protein is produced VrnH1 is allowed expressionVrnH1 is allowed expression Growth and differentiationGrowth and differentiation

Spring TypesSpring Types

Two genotypesTwo genotypes VrnH2 repressor gene is deleted VrnH2 repressor gene is deleted

No repressor for VrnH1 No repressor for VrnH1 Part of VrnH1 gene deletedPart of VrnH1 gene deleted

Nothing for VrnH2 to repressNothing for VrnH2 to repress

Spring plantedSpring planted No cold periodNo cold period No vernalizationNo vernalization

VrnH2

VrnH1

MeasurementMeasurement

How do you measure how long the plant How do you measure how long the plant grows?grows?

MethodsMethods Double ridgeDouble ridge stage of meristem development stage of meristem development Heading dateHeading date Final leaf number (FLN), (Fowler et al. 2006)Final leaf number (FLN), (Fowler et al. 2006)

Heading Date vs. Final Leaf Heading Date vs. Final Leaf NumberNumber

Heading date and Final leaf Number (FLN) are Heading date and Final leaf Number (FLN) are used to confirm genotypic and phenotypic used to confirm genotypic and phenotypic expectationsexpectations

Heading date can be influenced by vernalization Heading date can be influenced by vernalization requirement and maturity genesrequirement and maturity genes

Final leaf number (FLN) is also important an Final leaf number (FLN) is also important an important measurementimportant measurement

Experimental DesignExperimental Design

Heading date vs. FLN Heading date vs. FLN

Develop populations that segregate for Develop populations that segregate for winter types winter types (v1v1/V2V2, v1v1/V2v2)(v1v1/V2V2, v1v1/V2v2)

Collect data on populationsCollect data on populations Heading date (Szucs et al. 2006)Heading date (Szucs et al. 2006) FLNFLN

ObjectivesObjectives

Data Collection on FLN Data Collection on FLN Genotype Genotype (Szucs et al. 2007)(Szucs et al. 2007) Phenotype Phenotype

ParentsParents F1 progenyF1 progeny F2 progenyF2 progeny

Evaluate the dataEvaluate the data Heading date results (Szucs et al. 2007)Heading date results (Szucs et al. 2007) FLN resultsFLN results

Parents and CrossesParents and Crosses

Parents Parents CalicuchimaCalicuchima DicktooDicktoo Oregon Wolfe Barley Dominant (OWB-D)Oregon Wolfe Barley Dominant (OWB-D)

CrossesCrosses Dicktoo x OWB-DDicktoo x OWB-D Dicktoo x CalicuchimaDicktoo x Calicuchima

Crosses and HypothesesCrosses and HypothesesParent 1 Parent 2 F2

Dicktoo OWB –DExpect

segregation

Genotype Phenotype Genotype Phenotype for Vrn

v1v1/v2v2 No Vrn requirement

V1V1/V2V2 No Vrn requirement

requirement

Parent 1 Parent 2 F2

Dicktoo Calicuchima Expect segregation

Genotype Phenotype Genotype Phenotype for Vrn

v1v1/v2v2 No Vrn requirement

V1V1/V2V2 No Vrn requirement

requirement

Winter Genotypes:v1v1/V2V2V1v1/V2v2

Materials and MethodsMaterials and Methods

The F2 plants were grown in the greenhouse and not allowed a cold period for vernalization

FLN was measured on 93 F2 plants plus parents and F1 progeny Number of leaves on a the main tiller

ResultsResults

Analysis of FLNAnalysis of FLN Phenotypic Frequency DistributionsPhenotypic Frequency Distributions Mean Final Leaf Number of genotypesMean Final Leaf Number of genotypes Correlation between heading date and FLNCorrelation between heading date and FLN

Genotype Final Leaf Number Growth Habit VRNH1/VRNH2 Germplasm

Dicktoo 6.63 ± 0.86 Falcutative v1v1v2v2 Parent

OWB-D 3.25 ± 0.43 Spring V1V1/V2V2 Parent

Calicuchima 7.50 ± 0.5 Spring V1V1/V2V2 Parent

Dicktoo x OWB-D 5.50 ± 0.5 Spring V1v1V2v2 F1

Dicktoo x Calicuchima 6.50 ± 0.5 Spring V1v1V2v2 F1

Parents and F1 genotypesParents and F1 genotypes

Phenotypic DistributionPhenotypic DistributionHeading DateHeading Date

Dicktoo x OWB-DDicktoo x OWB-D

0

5

10

15

20

25

30

35

40

45

10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170

Days to flowering

Nu

mb

er o

f li

nes

OWB-D Dt

(Szucs et al. 2007)(Szucs et al. 2007)

Phenotypic DistributionPhenotypic DistributionFLNFLN

Dicktoo x OWB-D

0

5

10

15

20

25

30

35

1 2 3 4 5 6 7 8 9 10 11 12 13 14

Final Leaf Number

Lin

es

OWB-D Dt

Phenotypic DistributionPhenotypic DistributionHeading DateHeading Date

Dicktoo x CalicuchimaDicktoo x Calicuchima

0

5

10

15

20

25

30

35

40

45

10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170

Days to flowering

Nu

mb

er o

f li

nes

Dt Cb

(Szucs et al. 2007)(Szucs et al. 2007)

Phenotypic DistributionPhenotypic DistributionFLNFLN

Dicktoo x Calicuchima

0

5

10

15

20

25

30

35

40

1 2 3 4 5 6 7 8 9 10 11 12 13 14

Final Leaf Number

Lin

es

Dt Cb

Dicktoo x OWB-D

0

2

4

6

8

10

12

14

Genotype

Fin

al L

ea

f Nu

mb

er

Mean Final Leaf NumberMean Final Leaf Number

Dicktoo x Calicuchima

0

2

4

6

8

10

12

Genotype

Fin

al L

ea

f Nu

mb

er

Mean Final Leaf NumberMean Final Leaf Number

CorrelationCorrelation

Dicktoo x OWB-DDicktoo x OWB-D r = 0.94r = 0.94 P < 0.0001 (extremely significant)P < 0.0001 (extremely significant)

Dicktoo x CalicuchimaDicktoo x Calicuchima r = 0.8r = 0.8 P < 0.0001 (extremely significant)P < 0.0001 (extremely significant)

Results show a high correlation between Results show a high correlation between heading date and FLN dataheading date and FLN data

ConclusionsConclusionsResults show that genotypic and phenotypic Results show that genotypic and phenotypic FLN data matches with that of the Vrn gene FLN data matches with that of the Vrn gene modelmodel Results show significance for expected segregationResults show significance for expected segregation

Statistical analysis show that heading date and Statistical analysis show that heading date and FLN data are similarFLN data are similar High correlation between FLN and Heading Date High correlation between FLN and Heading Date

Final Leaf NumberFinal Leaf Number Smaller populationsSmaller populations Larger PopulationsLarger Populations

International and Local International and Local Research ExperienceResearch Experience

Comparison between Lledia, Spain Comparison between Lledia, Spain and Corvallis, Oregon, USAand Corvallis, Oregon, USA

OverviewOverview

Universidad de LleidaUniversidad de Lleida

Oregon State UniversityOregon State University

ComparisonComparison

SpainSpain

Olives

Grains

Goats

Citrus Fruits

History of Universidad de LleidaHistory of Universidad de Lleida

Second Oldest Spanish UniversitySecond Oldest Spanish University Created in 1300 by a charter Created in 1300 by a charter King of Aragon, James II King of Aragon, James II

Continued with vitality until 1717Continued with vitality until 1717 Royal LawRoyal Law Banning universities in Catalonia Banning universities in Catalonia SeminarySeminary

Officially reopened in 1968Officially reopened in 1968 New buildings addedNew buildings added University is center of townUniversity is center of town

Universidad de LleidaUniversidad de Lleida

Current UniversityCurrent University 10,000 students10,000 students Buildings located all over the Buildings located all over the

citycity Students stay in their Students stay in their

buildingbuilding Students live at homeStudents live at home Importance of Agriculture Importance of Agriculture

Region’s economyRegion’s economy

Many studies related Many studies related

University of LleidaUniversity of Lleida

Research ExperienceResearch Experience

International perspective on plant genetics International perspective on plant genetics researchresearch Laboratory of Dr. Ignacio RomagosaLaboratory of Dr. Ignacio Romagosa Researched root growth and structure of Researched root growth and structure of

barley barley Root growth and structureRoot growth and structure

Important in the role of drought, stress and nutrient Important in the role of drought, stress and nutrient uptake efficiencyuptake efficiency

Barley cultivars, OWBBarley cultivars, OWB

Overview of ProjectOverview of Project

Research ExperienceResearch Experience FundingFunding

Techniques and technology usedTechniques and technology used

Growth chambers in labGrowth chambers in lab

Basic PCR technologiesBasic PCR technologies

Membership in the EU has allowed greater access to fundingMembership in the EU has allowed greater access to funding Work ethic and cultureWork ethic and culture

Laid back atmosphereLaid back atmosphere

Very friendlyVery friendly

Coffee breaks and lunch breaksCoffee breaks and lunch breaks

Oregon State Oregon State UniversityUniversity

History of Oregon State UniversityHistory of Oregon State University

Settlers from the Oregon TrailSettlers from the Oregon Trail Corvallis – “Heart of the Valley”Corvallis – “Heart of the Valley”

Corvallis College – 1856Corvallis College – 1856 First class - 1870First class - 1870

3 Students graduated: 2 men, 1 woman3 Students graduated: 2 men, 1 woman

$10.00 per term for tuition$10.00 per term for tuition

1883 – First Department of Agriculture1883 – First Department of Agriculture

1885 – State of Oregon, control of College1885 – State of Oregon, control of College

History of Oregon State UniversityHistory of Oregon State University

1900 – Oregon Agricultural College1900 – Oregon Agricultural College Large growth of collegeLarge growth of college Addition of new departmentsAddition of new departments

1961- Oregon State University1961- Oregon State University

Sea, Land, Space and Sun grantSea, Land, Space and Sun grant

First Graduation in Gill Coliseum 1950

Linus Pauling Nobel Prize Winner

Oregon State UniversityOregon State UniversityOSU currentlyOSU currently 19,000 students 19,000 students Over 200 academic degree programs Over 200 academic degree programs Thirteen collegesThirteen colleges ResearchResearch

$194 million in funded research last year$194 million in funded research last year

Oregon State UniversityOregon State University

Research ExperienceResearch Experience Dr. Patrick HayesDr. Patrick Hayes Research conducted with Barley Research conducted with Barley

Stripe Rust resistance in BarleyStripe Rust resistance in Barley

Field, greenhouse, and lab workField, greenhouse, and lab work FundingFunding

Top technology and tools in the labTop technology and tools in the lab

Large fieldwork and research projectsLarge fieldwork and research projects Work ethic and cultureWork ethic and culture

Very motivated and hard workingVery motivated and hard working

Helpful and friendlyHelpful and friendly

ComparisonComparison

OverviewOverview Perspectives on researchPerspectives on research FundingFunding

More technology to aid in researching barley More technology to aid in researching barley geneticsgenetics

Larger impact with local, national, and international Larger impact with local, national, and international researchresearch

Both experiences aided me during my college Both experiences aided me during my college careercareer

Questions?Questions?