Rebecca E. Blanton 1 , Robert E. Jenkins 2 , and Henry L. Bart, Jr. 1

1. Morphological Variation Among Populations of the Federally Endangered Duskytail Darter, Etheostoma (Catonotus) percnurum Rebecca E. Blanton1, Robert E. Jenkins2, and Henry L. Bart, Jr.1 1Dept. Ecology and Evolutionary Biology, Tulane University, New Orleans, LA 2Dept. Biology, Roanoke College, Salem, VA

2. Taxonomic History • Formerly Etheostoma flabellare, Fantail Darter • First recognized as distinct species • R. E. Jenkins 1976 • Diagnosed and described by Jenkins in 1994 • Freshwater Fishes of Virginia (Jenkins and Burkhead 1994) • E. percnurum – Duskytail Darter • Reduced caudal barring relative to other closely related taxa E. flabellare E. percnurum

3. Classification: Catonotus E. squamiceps group “spottails” 10 spp. E. flabellare group “fantails” 3 spp. E. virgatum group “barcheeks” 7 spp. “Egg Mimics” pronounced on second dorsal Enlarged golden knobs (egg mimics) on first dorsal Blue, red in male fins Bar pattern (egg mimics) on cheek 3 black spots on caudal peduncle Nape, prepectoral area unscaled Female inverted for entire spawning event Female with broad, flat, non-bifurcate genital papilla Egg clustering

4. Distribution Modified from Jenkins and Burkhead 1994 = historically extirpated population

5. Morphological Variation • Noted by Jenkins(Jenkins and Burkhead 1994) • Differences in squamation and color • Substantial between Copper Cr. and others • Examined further by Eisenhour and Burr (2000) • Found differences in: • squamation and shape • egg and ova counts • nest size preference • Hypothesized Big South Fork population was independent evolutionary unit

6. Question • Noted morphological variation along with high degree of isolation of populations • Suggests each is likely on its own evolutionary trajectory • Are disjunct populations morphologically diagnosable, warranting species-level recognition??

7. Objectives • Examine morphological variation: • Determine taxonomic status of each population • Describe species that warrant recognition • Address conservation issues that may result from recognition of additional taxa

8. Methods

9. Methods • Specimens • Borrowed from museums and universities • Variation examined using: • Meristics • Morphometrics • Pigmentation • Intra and Inter-population variation

10. Methods • MERISTICS: • 26 counts • Largely followed Hubbs and Lagler (1958), Page (1983), and Jenkins and Burkhead (1994) • 194 specimens examined from 6 populations and 2 drainages • Males and females > 30 mm SL • Tested for sexual dimorphism • Males and females combined

11. Methods • MERISTIC DATA ANALYSIS: • Univariate: Frequency distributions • Multivariate: PCA (SYSTAT 5.2.1) • 16 of 26 variables used for all individuals • Factor scores plotted in Cricket Graph III • Variables with component loadings >.50 were considered most important

12. Methods • MORPHOMETRICS • Comparisons of body shape based on: • Truss protocol (Bookstein et al., 1985) using 19 interlandmark distances • 12 additional length/depth measurements (Hubbs and Lagler 1958) E. percnurum_Copper Cr.

13. Methods • MORPHOMETRIC ANALYSIS • Multivariate sheared PCA(Swofford; SAS 8.2) • 12 variables and 123 individuals • Univariate comparisons • Measurements standardized with Standard Length • Males and females analyzed separately • Nuptial and non-nuptial males examined separately • Included individuals > 35mm SL

14. Methods • PIGMENTATION: • Limited data due to lack of nuptial and live individuals • Data examined for 4 pigmentation variables • Jenkins previously examined other color and pigmentation characters of nuptial males(Jenkins and Burkhead 1994) • ANALYSIS • Univariate (frequency distributions, means) • Descriptive comparisons

15. Results MERISTICS

16. PCA: Meristics Fewer: Pored Lateral Line Belly Scales Higher: Caudal Fin Rays PC 2 Higher: Transverse Scales Caudal Peduncle Scales Scales Along D1 Base

17. 100% 0% 50% Squamation Along D1 Big South Fk. Citico Cr. Abrams Cr. Copper Cr. Little R.

18. Results MORPHOMETRICS

19. ResultsMorphometrics • No variation between females from different populations • Substantial overlap between non-nuptial males from different populations • Nuptial males with high degree of body shape variation between populations

20. Sheared PCA: Nuptial Males Shorter: First Dorsal Height Second Dorsal Height Caudal Fin Length Longer: Pelvic insertion to Anal Fin insertion PC3 Longer: Pectoral Fin Length Anal Fin Height

21. Results PIGMENTATION

22. Means for Pigmentation Variables *No. caudal spots also examined but showed no variation

23. Other Pigmentation DifferencesLittle R. vs. Copper Cr. Nuptial Males • Pelvic fins best pigmented in Copper Cr. • Rays of D2 more prominently marbled in Little R. • Rays uniformly dusky in Copper Cr. • Tessellations on Caudal Fin more intense in Little R. • Tessellations lost in Copper Cr.

24. Conclusions • Substantial morphological variation among four extant populations • Status of extirpated populations unclear due to low number of existing specimens • Abram Creek and South Fork Holston River • Grouped with closest geographic counterparts

25. Conclusions • Do any populations warrant species recognition? • Little River E. percnurum diagnosed with % scale cover along D1 base • also show distinct variation in morphometrics, and pigmentation • Others diagnosed with combination of characters: • Citico Creek – meristics and pigmentation • Big South Fork – measurements • Data needed for additional nuptial males • Copper Creek –combination of characters

26. Conclusions • Each population should be recognized: • Independent evolutionary units/species • Conservation efforts should be structured to conserve all populations • not just to maintain diversity, but because all are important independent units in the E. percnurum lineage

27. Future Questions and Work • Examine additional nuptial males from Citico Creek and Big South Fork • Live and nuptial males for better pigmentation comparisons • Diagnose and Describe • Relationships within the E.flabellare group of Catonotus • Are E. percnurum spp. monophyletic? • Is there high levels of genetic structure within populations similar to that observed for its supposed sister taxon, E.flabellare? • If so, important information for proper management and conservation

28. Johnson Cr. 1 (40%) 4 (20%) 27 (20%) 28 (20%) Hap 2 Stoney Cr. 2 (100%) 22 rmi. 19 rmi. Hap 1 7 rmi. Hap 27 Ararat R. 5 (20%) 6 (60%) 7 (20%) Hap 28 Hap 6 Hap 7 Yadkin R. Hap 6 Hap 6 NC Hap 5 Hap 1 Hap 4 NC SC ND2 Haplotype Locality Relationships Etheostoma flabellare

29. Acknowledgements • Institutions for loan of specimens: • INHS • UMMZ • University of Tennessee • Tulane Museum Natural History • Southern Illinois University Carbondale • American Museum Natural History • National Museum Natural History • Tulane University for facilities and equipment

30. Questions

32. Classification Etheostoma percnurum Duskytail Darter • Order: Perciformes • Family: Percidae • Genus: Etheostoma • Subgenus: Catonotus

33. Habitat Big South Fork, TN Citico Creek, TN

34. Conservation Status • Upon Description: • Jenkins called for federal protection • Relict distribution • Extirpation of two populations • Threats to habitat and water quality • Listed 1993 as Federally Endangered (Biggins 1993) • Recovery Plan developed 1994(Biggins and Shute)

35. Means of variables that loaded heavily on sheared PC 3 *Measurements standardized using SL

36. ResultsMeristics • Copper Creek and Citico Creek E. percnurum can be distinguished from one another based on meristics - no overlap in variation • Both also show high degree of meristic differences from Little R. and Big South Fork individuals • Little River and Big South Fork individuals with high degree of meristic similarity • Little River diagnosible based on scalation along D1 base