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