1. Barley Genetics as a Model for International Collaborative Research Lauren Osborn Oregon State University Department of Crop and Soil Science Mentors: Dr. Patrick M. Hayes Dr. Peter Szucs Dr. Peter Szucs Dr. Tony Chen Dr. Tony Chen

2. Chapters Inheritance of Final Leaf Number in Dicktoo x Calicuchima and Dicktoo x OWB-D F 2 populations International and Local Research Experience Oregon Wolfe Barley

3. Inheritance of Final Leaf Number in Dicktoo x Calicuchima and Dicktoo x OWB-D F 2 populations

4. Barley Winter types Plants survive harsh winter temperatures Plants survive harsh winter temperatures Vegetative State Vegetative State Facultative Types Low temperature tolerant but do not require cold period Low temperature tolerant but do not require cold period Spring types Planted in the spring Planted in the spring No need to survive winter temperatures No need to survive winter temperatures

5. Vernalization Requirement Induction of flowering by exposure to low temperature No VRN VRN Spring Facultative Winter

6. Sensitivity Growth Habit Winter Hardiness SensitivityVernalizationRequirementSensitivity Short day Photoperiod Sensitivity Winter Winter+++/- Facultative+-+/- Spring---

7. Vernalization Requirement Photoperiod Sensitivity Low Temperature Tolerance Cereal Winter Hardiness Low Temperature Tolerance Photoperiod Sensitivity

8. Vernalization Two genes determine vernalization requirement in barley (Von Zitzewitz, 2005) VrnH1 VrnH1 VrnH2 VrnH2 Winter alleles at both loci need to be present for the plant to require vernalization

9. VrnH1 VrnH1 is located on chromosome 5H Involved in plant growth and development Expression Plant undergoes a vegetative to reproductive transition Plant undergoes a vegetative to reproductive transition Meristem differentiation and the plant growth cycle Meristem differentiation and the plant growth cycle

10. VrnH2 VrnH2 is located on chromosome 4H VrnH2 encodes a repressor Effects VrnH1 Effects VrnH1 Prevents its expression Prevents its expression Repressor is active VrnH1 is not expressed VrnH1 is not expressed Plant remains vegetative Plant remains vegetative

11. Vrn H2 Winter types Repressor prevents expression of VrnH1 Repressor prevents expression of VrnH1 Until a period of low temperature Until a period of low temperature Cold period achieved VrnH2 genes cease expression VrnH2 genes cease expression No repressor protein is produced No repressor protein is produced VrnH1 is allowed expression VrnH1 is allowed expression Growth and differentiation Growth and differentiation

12. Spring Types Two genotypes VrnH2 repressor gene is deleted VrnH2 repressor gene is deleted No repressor for VrnH1 Part of VrnH1 gene deleted Part of VrnH1 gene deleted Nothing for VrnH2 to repress Spring planted No cold period No cold period No vernalization No vernalization

13. VrnH2 VrnH1

14. Measurement How do you measure how long the plant grows? Methods Methods Double ridge stage of meristem development Double ridge stage of meristem development Heading date Heading date Final leaf number (FLN), (Fowler et al. 2006) Final leaf number (FLN), (Fowler et al. 2006)

15. Heading Date vs. Final Leaf Number Heading date and Final leaf Number (FLN) are used to confirm genotypic and phenotypic expectations Heading date can be influenced by vernalization requirement and maturity genes Final leaf number (FLN) is also important an important measurement

16. Experimental Design Heading date vs. FLN Develop populations that segregate for winter types (v1v1/V2V2, v1v1/V2v2) Collect data on populations Heading date (Szucs et al. 2006) Heading date (Szucs et al. 2006) FLN FLN

17. Objectives Data Collection on FLN Data Collection on FLN Genotype (Szucs et al. 2007) Phenotype Phenotype Parents Parents F1 progeny F1 progeny F2 progeny F2 progeny Evaluate the data Heading date results (Szucs et al. 2007) Heading date results (Szucs et al. 2007) FLN results FLN results

18. Parents and Crosses Parents Calicuchima Calicuchima Dicktoo Dicktoo Oregon Wolfe Barley Dominant (OWB-D) Oregon Wolfe Barley Dominant (OWB-D)Crosses Dicktoo x OWB-D Dicktoo x OWB-D Dicktoo x Calicuchima Dicktoo x Calicuchima

19. Crosses and Hypotheses Parent 1Parent 2F2F2 DicktooOWB –D Expect segregation GenotypePhenotypeGenotypePhenotypefor Vrn v1v1/v2v2No Vrn requirement V1V1/V2V2No Vrn requirement requirement Parent 1Parent 2F2F2 DicktooCalicuchimaExpect segregation GenotypePhenotypeGenotypePhenotypefor Vrn v1v1/v2v2No Vrn requirement V1V1/V2V2No Vrn requirement requirement Winter Genotypes: v1v1/V2V2 V1v1/V2v2

20. Materials and Methods The F 2 plants were grown in the greenhouse and not allowed a cold period for vernalization FLN was measured on 93 F 2 plants plus parents and F 1 progeny Number of leaves on a the main tiller

22. Results Analysis of FLN Phenotypic Frequency Distributions Phenotypic Frequency Distributions Mean Final Leaf Number of genotypes Mean Final Leaf Number of genotypes Correlation between heading date and FLN Correlation between heading date and FLN

23. GenotypeFinal Leaf NumberGrowth HabitVRNH1/VRNH2Germplasm Dicktoo6.63 ± 0.86Falcutativev1v1v2v2Parent OWB-D3.25 ± 0.43SpringV1V1/V2V2Parent Calicuchima7.50 ± 0.5SpringV1V1/V2V2Parent Dicktoo x OWB-D5.50 ± 0.5SpringV1v1V2v2F1F1 Dicktoo x Calicuchima6.50 ± 0.5SpringV1v1V2v2F1F1 Parents and F1 genotypes

24. Phenotypic Distribution Heading Date Dicktoo x OWB-D (Szucs et al. 2007)

25. Phenotypic Distribution FLN OWB-DDt

26. Phenotypic Distribution Heading Date Dicktoo x Calicuchima (Szucs et al. 2007)

27. Phenotypic Distribution FLN Cb

28. Mean Final Leaf Number

30. Correlation Dicktoo x OWB-D r = 0.94 r = 0.94 P < 0.0001 (extremely significant) P < 0.0001 (extremely significant) Dicktoo x Calicuchima r = 0.8 r = 0.8 P < 0.0001 (extremely significant) P < 0.0001 (extremely significant) Results show a high correlation between heading date and FLN data

31. Conclusions Results show that genotypic and phenotypic FLN data matches with that of the Vrn gene model Results show significance for expected segregation Results show significance for expected segregation Statistical analysis show that heading date and FLN data are similar High correlation between FLN and Heading Date High correlation between FLN and Heading Date Final Leaf Number Smaller populations Smaller populations Larger Populations Larger Populations

32. International and Local Research Experience Comparison between Lledia, Spain and Corvallis, Oregon, USA

33. Overview Universidad de Lleida Oregon State University Comparison

34. Spain

35. Olives

36. Grains Goats Citrus Fruits

37. History of Universidad de Lleida Second 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 1717 Royal Law Royal Law Banning universities in Catalonia Banning universities in Catalonia Seminary Seminary Officially reopened in 1968 New buildings added New buildings added University is center of town University is center of town

38. Universidad de Lleida Current University 10,000 students 10,000 students Buildings located all over the city Buildings located all over the city Students stay in their building Students stay in their building Students live at home Students live at home Importance of Agriculture Importance of Agriculture Region’s economy Region’s economy Many studies related Many studies related

39. University of Lleida Research Experience International perspective on plant genetics research Laboratory of Dr. Ignacio Romagosa Laboratory of Dr. Ignacio Romagosa Researched root growth and structure of barley Researched root growth and structure of barley Root growth and structure Root growth and structure Important in the role of drought, stress and nutrient uptake efficiency Important in the role of drought, stress and nutrient uptake efficiency Barley cultivars, OWB Barley cultivars, OWB

40. Overview of Project Research Experience Funding Funding Techniques and technology used Techniques and technology used Growth chambers in lab Growth chambers in lab Basic PCR technologies Basic PCR technologies Membership in the EU has allowed greater access to funding Work ethic and culture Work ethic and culture Laid back atmosphere Laid back atmosphere Very friendly Very friendly Coffee breaks and lunch breaks Coffee breaks and lunch breaks

41. Oregon State University

42. History of Oregon State University Settlers from the Oregon Trail Corvallis – “Heart of the Valley” Corvallis – “Heart of the Valley” Corvallis College – 1856 First class - 1870 First class - 1870 3 Students graduated: 2 men, 1 woman 3 Students graduated: 2 men, 1 woman $10.00 per term for tuition $10.00 per term for tuition 1883 – First Department of Agriculture 1885 – State of Oregon, control of College

43. History of Oregon State University 1900 – Oregon Agricultural College Large growth of college Large growth of college Addition of new departments Addition of new departments 1961- Oregon State University Sea, Land, Space and Sun grant

44. First Graduation in Gill Coliseum 1950

45. Linus Pauling Nobel Prize Winner

47. Oregon State University OSU currently 19,000 students 19,000 students Over 200 academic degree programs Over 200 academic degree programs Thirteen colleges Thirteen colleges Research Research $194 million in funded research last year $194 million in funded research last year

48. Oregon State University Research Experience Dr. Patrick Hayes Dr. Patrick Hayes Research conducted with Barley Research conducted with Barley Stripe Rust resistance in Barley Stripe Rust resistance in Barley Field, greenhouse, and lab work Field, greenhouse, and lab work Funding Funding Top technology and tools in the lab Large fieldwork and research projects Work ethic and culture Work ethic and culture Very motivated and hard working Very motivated and hard working Helpful and friendly Helpful and friendly

49. Comparison Overview Perspectives on research Perspectives on research Funding Funding More technology to aid in researching barley genetics Larger impact with local, national, and international research Both experiences aided me during my college career Both experiences aided me during my college career

50. Questions?