1. A Systematic Evaluation of Sisyrinchium hitchcockii Matt Groberg OSU and ODA Stephen Meyers, Dr. Robert Meinke (Advisors) S. hitchcockiiS. bellum S. idahoense

2. Conservation Status OrganizationStatus Natural Heritage Information Center List 1 NatureServeG1 Natural Heritage Network S2 USFWSSpecies of concern BLMSensitive species ODANot listed

3. BLM: Soggy Bottom Site S. hitchcockii

4. A Brief Taxonomic History: Hitchcock and Cronquist (1973) group S. hitchcockii, along with closely related species, into one species: Sisyrinchium angustifolium. Henderson (1976) divided groups of plants synonymous with S. angustifolium into eight different species including S. bellum, S. idahoense, and S. hitchcockii. –Previous taxonomic studies based on dried plants. –Characters in the flowers of Sisyrinchium are not well preserved in herbarium specimens. S. angustifolium S. bellum S. hitchcockii S. idahoense

5. Henderson explains that S. hitchcockii is morphologically unique in that it has: A short creeping rhizome The largest flowers ( tepals up to 20 mm long) The widest stems and leaves ( >3mm) Rhizome Tepal Leaf Stem

6. Definitions of a Species A member of a species can produce fertile offspring with another member of that species. –However, plants often self fertilize or hybridize with closely related species and produce fertile offspring. A group of organisms that are morphologically, cytologically, genetically, and distributionally very similar. –The species is on its own evolutionary track.

7. Speciation Allopatric Speciation: new species are formed when a geographic or ecological barrier between two populations prevents gene flow over a long period of time. Sympatric Speciation: new species are formed when internal barriers, such as polyploidy, reduce or prevent gene flow between the new species and the preserved species.

8. Polyploidy

9. Distribution S. hitchcockii S. bellum S. idahoense

10. Chromosome Counts (Henderson 1976) S. hitchcockii: n = 32 S. bellum: n = 16 S. idahoense: n = 32 or 48 Suggests Polyploidy present in all three

11. Self Compatibility (Henderson1976)

12. Cross Compatibility (Henderson1976)

13. Question and Hypotheses Q: Is S. hitchcockii a true species? –H0: S. hitchcockii is a larger version of S. bellum due to polyploidy, but is not genetically unique and therefore a not true species. –H1: S. hitchcockii is morphologically, cytologically, and genetically unique and therefore a true species.

14. Objectives Compare the morphology of S. bellum and S. idahoense to S. hitchcockii from fresh specimens and herbarium specimens to determine if there are differences not preserved in herbarium specimens.

15. Objectives Compare the morphology of S. bellum and S. idahoense to S. hitchcockii from fresh specimens and herbarium specimens to determine if there are differences not preserved in herbarium specimens. Confirm that there is polyploidy in chromosome numbers between all three species.

16. Objectives Compare the morphology of S. bellum and S. idahoense to S. hitchcockii from fresh specimens and herbarium specimens to determine if there are differences not preserved in herbarium specimens. Confirm that there is polyploidy in chromosome numbers between all three species. Analyze genetic information from all three species to determine if S. hitchcockii is unique and to suggest a possible phylogenetic tree.

17. Objectives Compare the morphology of S. bellum and S. idahoense to S. hitchcockii from fresh specimens and herbarium specimens to determine if there are differences not preserved in herbarium specimens. Confirm that there is polyploidy in chromosome numbers between all three species. Analyze genetic information from all three species to determine if S. hitchcockii is unique and to suggest a possible phylogenetic tree. Determine if S. hitchcockii is a true species that should be monitored.

18. Objectives Compare the morphology of S. bellum and S. idahoense to S. hitchcockii from fresh specimens and herbarium specimens to determine if there are differences not preserved in herbarium specimens. Confirm that there is polyploidy in chromosome numbers between all three species. Analyze genetic information from all three species to determine if S. hitchcockii is unique and to suggest a possible phylogenetic tree. Determine if S. hitchcockii is a true species that should be monitored. Develop a key for identification in the field.

19. Materials and Methods Morphology: key traits were measured in each species and then statistically compared using morphometric analysis.

20. Materials and Methods Morphology: key traits were measured in each species and then statistically evaluated using morphometric analysis. Cytology: cytometry from crushed leaves was made with Partec GmbH ploidy analyzer ( at OSU seed lab.)

21. Materials and Methods Morphology: key traits were measured in each species and then statistically compared using morphometric analysis. Cytology: cytometry from crushed leaves was made with Partec GmbH ploidy analyzer ( at OSU seed lab.) Genetics: DNA was extracted from the leaves. Polymerase chain reaction (PCR) was used to amplify nuclear non-coding DNA (nrITS) and chloroplast DNA (matK). Phylogenetic tree was created using Bayesian inference.

22. Polymerase Chain Reaction

23. Reason for Using Non-Coding DNA Coding DNA Non-Coding DNA

24. Results and Discussion

25. Morphology S. hitchcockii Outer tepals: l:w ratio >2.5; length usually >15mm and up to 20mm; elliptic and slightly wider at the top Yellow eyeYellow eye: yellow eye small if present. Filaments: dark purple and >6mm long. S. bellum Outer tepals: l:w ratio <2.5; length <13mm long; egg shaped, wider at the top; topside glossy and underside lighter than topside. Yellow eyeYellow eye: yellow eye robust. Filaments: yellow to purple and < 6mm long. S. Idahoense Outer tepals: l:w ratio >2.5, usually >13mm and up to 20mm in some varieties; oblong to slightly wider at the top. Yellow eyeYellow eye: yellow eye present. Filaments: yellow-tan to tan-purple and < 6mm long. Outer tepals Fused filaments Yelloweye Yellow eye

27. Cytology Chromosome counts: –S. hitchcockii: n = 32 –S. bellum:n = 16 –S. idahoense:n = 64

29. Conclusion S. hitchcockii is unique:

30. Conclusion S. hitchcockii is unique: 1.Morphologically

31. Conclusion S. hitchcockii is unique: 1.Morphologically 2.Cytologically

32. Conclusion S. hitchcockii is unique: 1.Morphologically 2.Cytologically 3.Genetically

33. Acknowledgements Stephen Meyers Kelly Amsberry OSU and UC herbaria Dr. Robert Meinke Rebecca Currin Dr. Kevin Ahern & HHMI Dr. Lynda Ciufetti Dr. Lisa Karst Jaworski Scholarship