A Systematic Evaluation of Sisyrinchium hitchcockii Matt Groberg OSU and ODA Stephen Meyers, Dr. Robert Meinke (Advisors) S. hitchcockii S. bellum S. idahoense
Feb 24, 2016
A Systematic Evaluation of Sisyrinchium hitchcockii
Matt GrobergOSU and ODA
Stephen Meyers, Dr. Robert Meinke (Advisors)
S. hitchcockii S. bellum S. idahoense
Conservation StatusOrganization StatusNatural Heritage Information Center
List 1
NatureServe G1
Natural Heritage Network
S2
USFWS Species of concern
BLM Sensitive species
ODA Not listed
BLM: Soggy Bottom Site
S. hitchcockii
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.
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
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.
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.
Polyploidy
Distribution
S. hitchcockiiS. bellumS. idahoense
Chromosome Counts (Henderson 1976)
• S. hitchcockii: n = 32• S. bellum: n = 16• S. idahoense: n = 32 or 48
• Suggests Polyploidy present in all three
Self Compatibility (Henderson1976)
Cross Compatibility (Henderson1976)
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.
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.
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.
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.
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.
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.
Materials and Methods
• Morphology: key traits were measured in each species and then statistically compared using morphometric analysis.
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.)
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.
Polymerase Chain Reaction
Reason for Using Non-Coding DNA
Coding DNA Non-Coding DNA
Results and Discussion
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 eye: yellow eye small if present.
• Filaments: dark purple and >6mm long.
S. bellumOuter tepals: l:w ratio <2.5; length <13mm long; egg shaped,wider at the top; topside glossy andunderside lighter than topside.
• Yellow eye: yellow eye robust.
• Filaments: yellow to purple and < 6mm long.
S. IdahoenseOuter tepals: l:w ratio >2.5, usually>13mm and up to 20mm in some varieties; oblong to slightly wider at the top.
• Yellow eye: yellow eye present.
• Filaments: yellow-tan to tan-purple and < 6mm long.
Outer tepals
Fused filamentsYellow eye
Plot of Discriminant Functions
-7 -5 -3 -1 1 3 5
Function 1
-3.2
-1.2
0.8
2.8
4.8
Func
tion
2
speciesS. bellum S. hitchcockii S. idahoense Centroids
Cytology
• Chromosome counts:– S. hitchcockii: n = 32– S. bellum: n = 16– S. idahoense: n = 64
ConclusionS. hitchcockii is unique:
ConclusionPlot of Discriminant Functions
-7 -5 -3 -1 1 3 5
Function 1
-3.2
-1.2
0.8
2.8
4.8
Func
tion
2
speciesS. bellum S. hitchcockii S. idahoense Centroids
S. hitchcockii is unique:1. Morphologically
ConclusionPlot of Discriminant Functions
-7 -5 -3 -1 1 3 5
Function 1
-3.2
-1.2
0.8
2.8
4.8
Func
tion
2
speciesS. bellum S. hitchcockii S. idahoense Centroids
S. hitchcockii is unique:1. Morphologically2. Cytologically
ConclusionPlot of Discriminant Functions
-7 -5 -3 -1 1 3 5
Function 1
-3.2
-1.2
0.8
2.8
4.8
Func
tion
2
speciesS. bellum S. hitchcockii S. idahoense Centroids
S. hitchcockii is unique:1. Morphologically2. Cytologically3. Genetically
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