Conservation Genetics of the Blueface Darter (Etheostoma sp. cf. zonistium), a rare

undescribed fish in northwest Alabama

Kenny Jones Brook L. Fluker

Bernard R. Kuhajda

Distribution and Characteristics

Bandfin Darter, Etheostoma zonistium

Blueface Darter, Etheostoma cf. zonistium

Fish art: Joseph R. Tomelleri

AL

MS

TN

KY

Gulf Coastal

Plain

Cumberland Plateau

Bailey and Etnier (1988); Kuhajda and Mayden (upubl. data)

Blueface Darter description is forthcoming (Kuhajda and Mayden).Once formally recognized, conservation status will need to be determined.

Previous Work

Phylogenetic analysis of cytochrome b (mtDNA) data identified Blueface Darters as a monophyletic group

Sister to E. zonistium sp. group

Approximately 3% sequence divergence

Kuhajda and Fluker (2009)

An Isolated Distribution

Natural isolation: The Hubbard Creek population is potentially isolated from other populations via natural isolation.

Reservoir-induced isolation: The Upper Bear Creek systems are possibly isolated due to the construction of the Upper Bear Cr. Reservoir (1978). Extant

Possibly extirpatedKuhajda and Mayden (2002)

Sipsey Fork

Little

Bear C

r.

Bear Cr.

Bear C

r.

Turke

y Cr.

Tennessee RiverDrainage

Black Warrior River Drainage (Mobile Basin)

Hubba

rd Cr.

Sipsey ForkUpper Bear

Cr. Reservoir

10.2 km

5.4 km

QuestionsI. Is there significant genetic structure between

Blueface and Bandfin Darters?II. Does the Blueface Darter have reduced

genetic variation compared to the Bandfin?III. Is there significant genetic structure within

the Blueface Darter across the Tennessee/Black Warrior divide?

IV. Does the Black Warrior population of Blueface Darter have reduced genetic variation compared to the Tennessee River drainage populations?

Methods: Sample collection

Upper BearCr.

Reservoir2

ALMS

TN

KY

Little

Bear C

r.

Bear Cr.

Bear

Cr.

Turke

y Cr.

Tennessee RiverDrainage

Black Warrior River Drainage (Mobile Basin)

Hubbard Cr.1

3

5

Sipsey Fork

Upper BearCr. Reservoir 2

6 478

9

Kinlock Falls, Hubbard Creek

20-30 individuals per site (seine) Fin-clipped (95% EtOH) Photo-vouchered or formalin preserved

specimens Extracted DNA using Qiagen DNeasy Kit

Methods: Microsatellite DNA analyses Genotyped 187 individuals for 8

microsatellite DNA loci: Esc132b, Esc68, Esc18, Esc26b (Gabel

et al. 2008) Eca46, Eca48, Eca49 (Tonnis 2006) EosD107 (Switzer et al. 2008)

Estimated genetic structure using AMOVA (ARLEQUIN) and STRUCTURE

Estimated genetic variation via allelic diversity and heterozygosity expected (ARLEQUIN)

Evaluated differences in genetic variation using non-parametric statistics (Kruskal-Wallis and Mann-Whitney U tests)

Structure v2.3.4

ResultsUpper Bear

Cr. Reservoir

2

ALMS

TN

KY

Little

Bear C

r.

Bear Cr.

Bear

Cr.

Turke

y Cr.

Tennessee RiverDrainage

Black Warrior River Drainage (Mobile Basin)

Hubbard Cr.1

3

4

Sipsey Fork

Upper BearCr. Reservoir 2

6 578

9

Results

1 63 4

Blueface Darter Bandfin Darter

2 5 7 8 9

FCT= 0.02, and an overall P-value of 0.03

1 2 3 4 5 6 7 8 9

Hubbard Above --Hubbard Below 0.003* --Bear Cr. (cf. zon) 0.098 0.066 --Little Bear Cr. 0.146 0.109 0.060 --

Turkey Cr. 0.079 0.073 0.022* 0.109 --

Big Sandy 0.070 0.041 0.033 0.067 0.064 --

Birdsong 0.073 0.056 0.034 0.076 0.070 0.023 --

Hatchie 0.247 0.228 0.205 0.234 0.247 0.176 0.155 --Bear Cr. (E. zon) 0.168 0.140 0.119 0.162 0.158 0.090 0.104 0.246 --

Results cont.

Blueface DartersBandfin Darters

Results

0

2

4

6

8

10

12

14

16

18

Blueface darters show a 23% reduction in genetic variation compared to Bandfin, but not significant (P = 0.084)

Mea

n Al

lele

s Per

Lo

cus

ResultsUpper Bear

Cr. Reservoir

2

ALMS

TN

KY

Little

Bear C

r.

Bear Cr.

Bear

Cr.

Turke

y Cr.

Tennessee RiverDrainage

Black Warrior River Drainage (Mobile Basin)

Hubbard Cr.1

3

4

Sipsey Fork

Upper BearCr. Reservoir 2

6 578

9

Results

1 63 4

Blueface Darter Bandfin Darter

2 5 7 8 9

1 2 3 4 5

Hubbard Above --

Hubbard Below 0.003* --

Bear Cr. (cf. zon) 0.098 0.066 --

Little Bear Cr. 0.146 0.109 0.060 --

Turkey Cr. 0.079 0.073 0.022* 0.109 --

Results cont.

Mea

n Al

lele

s Pe

r Lo

cus

Genetic diversity differed between Blueface populations (P=0.018). Turkey Creek lower than all others.

Hubbard Above Hubbard Below Bear Cr. Little Bear Cr Turkey Cr.0

5

10

15

20

25

Results

ConclusionsBlueface and Bandfin darters are differentiated with both

microsatellite and mtDNA. Species validity is supported with genetic data, although microsatellite data indicate retention of ancestral alleles in the Bear Creek population.

Overall, Blueface Darters do not have lower have lower genetic variation compared to the Bandfin Darter, even though allelic diversity was 23% percent lower and was a non-significant value.

Populations of the Blueface darter are isolated across the Tennessee/Black Warrior divide.

Possible reservoir fragmentation effect seen in Little Bear Creek, i.e. significant genetic structure and lower genetic variation compared to Bear Creek.

Conservation implications After being formally

described, the results generated from this study can inform state and federal agencies about the conservation needs of the Blueface darter

Conservation units Hubbard Creek Bear Creek system

Conservation priorities1. Turkey Creek 2. Hubbard Creek3. Little Bear Creek4. Bear Creek (source for

PTRA)

Acknowledgements

Funding Arkansas State University Faculty

Research Award Committee (FRAC). Tennessee Aquarium Conservation

Institute. Field and Lab Assistance

Alexandra Hook Taylor Lee Brittany McCall Dave Neely

Collecting Permits Alabama Dept. of Conservation and

Natural Resources Tennessee Wildlife Resources

Agency

Closing Statement