till .K4 T7 v. 65 no. 2 Fal 2004 JOURNAL KENTUCKY ACADEMY OF SCIENCE Official Publication of the Academy Volume 65 Number 2 Fall 2004 The Kentucky Academy of Science Founded S May 1914 Governing Bourn 2004 Executive Committee 2004 President: Robert W. Kingsolver, Kentucky Wcslcvan CoUege/kingsoI@kwc.cdu President Elect: Bruce Mattingly, Morehead State University/b. mattin@morehead-st.edu Vice President: Miriam Sleinitz-Kannan, Northern Kentucky Universily/kannan@nku.cdu Past President: Robert J. Barney, Kentucky State University/rbamey@kysu.edu Secretary: Dawn Anderson, Berea College/dawn-anderson@berea.edu Treasurer: Kenneth Crawford, Western Kentucky University/kenneth. crawford@wku.edu Executive Secretary (ex officio): Donald Frazier, University of Kentucky/dfrazie@pop. uky.edu Editor, JOURNAL (ex officio): John VV. Thieret, Northern Kentucky' Universitv/thieretj@exchange. nku.edu DmSIOX AND AT-LARCE REPRESENTATIVES Physical Sciences (2004): Jennifer Muzyka, Centre ColIege/muzyka@centre.edu Biological Sciences (2005): Thomas Rambo, Northern Kentucky Universitv/rambot@nku.edu At-Large (2005): Ralph Thompson, Berea College/ralph_thompson@berea.edu Social Sciences (2006): Darid Hogan, Northern Kentucky University/hogan@nku.edu Physical Sciences (2006): Mark Blankenbuehler, Morehead State University/blanken@morehead-st.edu Program Coordinator (ex officio): Robert O. Creek, Eastern Kentucky University/rcreek@chpl.net Director, Junior Academy of Science (ex officio): Elizabeth K. 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INSTITUTIONAL AFFILIATES Fellow University of Kentucky University of Louisville Sustaining Member Campbells ville University Kentucky State University Morehead State University Murray State University Northern Kentucky University Western Kentucky University Member Asbury College Bellarmine University Berea College Centre College Cumberland College Eastern Kentucky Univerity Kentucky Wesleyan College Madisonville Community College West Kentucky Community & Technical College Associate Member Pikeville College Transylvania University INDUSTRIAL AFFILIATES Associate Patron Touchstone Energy Associate Member Hoffman Environmental Research Institute Third Rock Consultants Wood Hudson Cancer Research Lab Figure 1. Fatoua villosa. a. Whole plant, b. Inflorescence, c. Staminate flower, d. Staminate flower, long section, e. Floral diagram of staminate flower, f. Pistillate flower, g. Pistillate flower, long section, h. Floral diagram of pistillate flower, i. Fruit, j. Seed. Bar = 1.5 cm for a; 0.5 mm for b; 1 mm for c, d, f, g-j: Drawings by Priscilla Fawcett; taken from Correll and Correll 1982; used by permission of the publisher. JOURNAL OF THE KENTUCKY ACADEMY OF SCIENCE ISSN 1098-7096 Continuation of Transactions of the Kentucky Academy of Science Volume 65 Fall 2004 Number 2 J. Ky. Acad. Sci. 65(2):67-75. 20(14. Spread of Fatoua villosa (Mulberry Weed; Moraceae) in North America Michael A. Vincent W. S. Turrell Herbarium (MU), Department of Botany, Miami University, Oxford, Ohio 4.5056 ABSTRACT Fatoua villosa (mulberry weed, Moraceae) is documented from 28 states and die District of Columbia in die United States. The species has spread through the horticultural trade to greenhouses and nurseries, from which it has escaped into gardens, lawns, and ruderal areas. A small percentage of the collections examined were from more natural settings in open forested sites or along streams. The species is illustrated and described, and maps are provided showing its spread in the 50 years since it was first collected in this country. First reports are provided for mulberry weed in die District of Columbia, Illinois, Minnesota, Oregon, Pennsylvania, and Wisconsin. INTRODUCTION Forty years ago, Thieret ( 1964) reported the discovery of a new weedy plant species for North America, Fatoua villosa (Thunb.) Na- kai, commonly called "mulberry weed," "hairy crabweed," or "kuwa-kusa," a member of the Moraceae (mulberry family). Thieret cited in- formation provided by Dr. Joseph Ewan, of New Orleans, that the species had been known in diat city for about 15 years. Since that time, the species has spread across North America, and literature reports exist for its presence in 26 states in the United States. The genus Fatoua was described by Gau- dichaud (Freycinet 1830). Two or three spe- cies of the genus are native to Australasia (Chew 19S9; Hutchinson 1967; Rohwer and Berg 1993; Zhou and Gilbert 2003) and an ad- ditional species is found in Madagascar (Berg 1977; Leandri 1948). Fatoua villosa (Figure 1) was first described by Thunberg (1784) as Ur- tica villosa (Urticaceae); the epithet was trans- ferred to the genus Fatoua by Nakai (1927). Synonyms include Urtica japonica Thunberg (1784), Fatoua japonica Blume (1856), and Boehmeriopsis pallida Komarov (1901). Mi- quel (1869) published the only monograph of the genus to date, treating it as a member of the Urticaceae. The first treatment of Fatoua as a member of Moraceae was by Gaudichaud (Freycinet 1830), who described two addition- al possible synonyms, F. pilosa and F. aspera. Placement of the genus in Moraceae is sup- ported by Yamazaki (1982), who showed that seed development patterns in Fatoua follow those found in other Moraceae and is unlike that of Urticaceae. Mulberry weed is native to China (Li 1986), [apan (Ohwi 1965), Korea (Komarov 1901; Lee 1989), Okinawa (Walker 1976), Ryukyu Islands (Walker 1976), Taiwan (Liu and Liao 1976), and Tonkin (Reed 1977). In addition to reports from the continental United States, which will be discussed below, mulberry weed has been reported as an introduced weed in the Bahamas (Correll and Correll 1984), Cor- sica (Jeanmonod 2000), Finland (Kuitunen and Lahtonen 1994). Hawaii (Yatskievych and Raveill 2001), and Puerto Rico (Liogier 1997; Liogier and Martorell 2000). In its native range, mulberry weed is considered weedy in fi,S [ournal of the Kentuck) Academ) of Science 65(2) cultivated or grassy fields, along roadsides, on trailsides, in open woods, and in rocky areas (Holm et al. 1979; Ohwi 1965; Randall 2002; Walker 1976; Zhou and Gilbert 2003). As an introduced weed, it has been found in green- houses, nurseries, flowerbeds, potted plants, and botanical gardens; a few records are from woodlands, riverbeds, and forest edges and fencerows. Fat ou a villosa is an herbaceous, erect, tap- rooted annual with colorless sap without latex. Stems of the species are 10-90 cm in height, may be simple to much branched, and range in color from green to maroon red; they are thinly to densely short hairy. In general aspect, die species greatly resembles taxa of Urtica- ceae and, indeed, when first encountered is often mistakenly identified as a member of that family. Its leaves are alternate, thinly sca- brous, broadly ovate to lanceolate, 3-10 X 1- 5 cm, tootiied, cordate to truncate at the base, acute to acuminate at the apex; petioles range from 0.5-6 cm; stipules are small, free, and early-deciduous; venation in the leaves is pal- mate to pinnate. Punctate cystolitiis, com- posed of calcium carbonate, are present in die leaves. Inflorescences are axillary, condensed cymes that are often nearly capitate, 4—7 mm wide, on peduncles that vary from very short to 2-3 cm in length; each inflorescence is sub- tended bv a small bract. Plants are monoe- cious, with staminate and pistillate flowers in each inflorescence; inflorescences lower on the stem contain predominately pistillate flow- ers; die further up the stem the inflorescence is positioned, the larger the percentage of sta- minate flowers. Among the fertile pistillate flowers can be found staminate flowers with morphologically distinct pistils (pistillodes) without ovules. The calyx is 4-lobed, pubes- cent, yellowish in staminate flowers, and green in pistillate flowers; die corolla is absent. Functional pistils are bicarpellate and uniloc- ular; the style is lateral and appears un- branched, but has a very small aborted stvle branch positioned at die base of the larger functional style; there is a single pendulous ovule. Staminate flowers contain four exserted stamens positioned alternately widi the calyx lobes. Fruits are achenes, three-angled to nearly globose to flattened, 1—1.1 mm long, buff to dark brown, with whitish raised ridges; they are mosdy dropped near the mother plant, but are also explosively expelled up to 4' — and probably farther, especially from plants growing as weeds in flower baskets hanging over greenhouse benches. Seeds have a small embrvo, with endosperm, i Description compiled from specimens and the following sources: Miller and Wood 2003; Neal 1998; Reed 1977; Rohwer and Berg 1993; Sanders 1996; Thieret 1964; Wu and Kuo-Huang 1997: Wunderlin 1997; Yamazaki 1982; Yatskiewch and Raveill 2001; Zhou and Gilbert 2003). Known chromosome counts are n = 13 (Kon- do and Miller 1973) and In = 26 Li (1986). Plants flower from July dirough October in outdoor settings in the north, August through November in the south, and through the sum- mer and winter in deep south and greenhouse settings. Swain and Downum (1989, 1990) showed that mulberrv weed contains biologically ac- tive compounds, furanocoumarins, diat are phototoxically active toward some test organ- isms, such as die bacterium Escherichia coli. Control of the species as a weed has been extensively studied by Penny and Neal (1999a, 1999b). Pre-emergence control is most effec- tively obtained by use of products containing ox\-flourfen and oxadiazon; dinitroaniline her- bicides differed in effectiveness. Penny and Neal (2003) showed that seed burial and mulch were 90% effective at inhibiting ger- mination, since germination requires light. Low or high temperatures may also affect ger- mination and development, since die species grew best at temperatures between 15-38°C (Penny 2000). Postemergence control is most effective using products containing paraquat, glyphosate, and glufosinate and perhaps also diquat or pelargonic acid (Pennv 2000). In North America, mulberry weed is re- ported bv Wunderlin (1997) from 17 states in the United States, and bv Kartesz and Mea- cham (1999) and the USDA Plants Database (USDA, NRCS 2004) for 18 states. Additional reports in the literature bring die number to 26 states (Table 1). No reports are known of the species in Canada. It appears that die spe- cies is being spread dirough the horticulture industry, since many earlv reports are of die species as a weed in greenhouses, from which it presumably spreads by means of bedding plants, nursery stock, potting soil, or bedding mulch (Miller and Wood 2003; Pennv 2000; Spread ol Fatoua villosa in North America — Vincent m Table I. States in the United Stales from which Fatoua oillosa lias been reported in published literature or observed as herbarium specimens, Listed are the source citations, the year ol the earliest known collection examined during the course of this study, ami the number ol counties/parishes from which (lie species is reported (specimens ami undoi umented literature reports). Literature sourcc(s) Earliest ...II-. Number <>t ties parishes Alabama Arkansas California District ol Columbia Florida Georgia Illinois Indiana Iowa Kentucky Louisiana Maryland Massachusetts Michigan Minnesota Mississippi Missouri New York North Carolina Ohio Oklahoma Oregon Pennsylvania South Carolina Tennessee Texas Utah Virginia Washington West Virginia Wisconsin M.issr\ L975 Smith 1994; Sundell 1986 Ilrusa el al. 2002: Randall 1997: Sanders L996; Baldwin ct al. 2004 DuQuesna) 197-1: Massey 1975: Wunderlin and Hansen '2004 Jones and Coile L988; Massey 1975 Maxwell and Thomas 2003: Wunderlin 1997 Cusick 2002 Brownie and Athev 1992; Taylor 1994 MacRoberts 19S9; Massey 1975; Thieret 1964; Thomas et al. 1980 Wunderlin 1997 Miller and Wood 2003 Reznicek 2001 Carter et al. 1990; Massev 1975 Wunderlin 1997; Yatskievych and Raveill 2001 Miller and Wood 2003 Massey 1975; Neal 1998 Vincent 1993 Taylor et al. 1996; Taylor and Taylor 1981 Anonymous 2004b; Wunderlin 1997 Krai 1981; Anonymous 2004c Jones et al. 1997; Lipscomb 1984 Welsh et al. 2003 Wright 198S Anonymous 1996; Anonymous 2004d Wunderlin 1997 1967 S L985 3 1983 7 1994 1 1970 17 1965 11 1982 2 1995 1 2000 1 1983 6 1950 IS 1981 4 1994* 1 2001 1 199S 1 1972 9 1972 7 2002 1 1973 7 1979 11 1979 7 2000 1 1989 1 1975 7 1970 4 1974 9 1997 1 19S7 4 1995* 1 2002 * = not seen. Penny and Neal 1999a, 1999b; Sanders 1996; Taylor 1994; Taylor et al. 1996; Vincent 1993; Welsh et al. 2003; Wunderlin 1997; Yatskiev- ych and Raveill 2001). Pennv (2000) and Pen- ny and Neal (1999a, 1999b) report 50% of nurseries surveyed in 1997 and 75% of nurs- eries surveyed in 1998 were infested by this weedv species. One or two generations can oc- cur in a growing season (Penny 2000; Pennv and Neal 1999a! 1999b). Tl lis study was undertaken to determine the extent to which Fatoua cillosa has spread in North America since its introduction, and if any pattern can be discerned regarding its spread. MATERIALS AND METHODS In addition to field work to find the species, herbarium specimens were examined from the following herbaria (acronyms from Holmgren 2004): A, AUA. BALT, BAYLU, BH. BHO. BKL, BRIT, C, CDA, CLEMS, CM. DAN". DOY, F. FTG, GA. GH. GMUF, ID. ILL, ILLS, ISC. JEF, KNK. LAF. LSU, MICH. MIN, MISS, MISSA, MO, MONTU, MU. NA, NBYC, NCU, NHA, NLU. NO, NY, OCLA, OKL, OS, OSH, RM, SMU. TENN. TEX. UAM, UCR, UNA, UNLY. URY. US, USCH. USF VDB, VPI, VT, WIS. WSI WTU, WVA. 70 journal "I the Kentucky Acad Science 65(2) >V5>* Figure 2. Distribution of Fatoua villosa in North America from its initial discovery in Louisiana to the present. Each square (■) represents the first report of the species for a particular count)'. Dots (•) represent these first reports carried over from previous map(s). a. 1950-1969. b. 1970-1979. c. 1980-1989. d. 1990-2004. RESULTS AND DISCUSSION A total of 495 herbarium specimens were examined during the course of this study. Col- lections were seen from 134 counties in 28 states, and the District of Columbia (Appendix 1). An additional 21 counties are listed in lit- erature reports for which no voucher docu- mentation was found. Of the counties repre- sented by specimens, 17 were represented only as greenhouse weeds and not by speci- mens from sites out-of-doors. The earliest known specimen of mulberry weed from North America is from New Or- leans, Orleans Parish, Louisiana, collected in 1950 (G. P. DeWolfs.n. [NO]). Table 1 shows the dates of the earliest known specimen from each state, as well as the number of counties/ parishes from which die species is known for each state. The state with the largest number of counties/parishes from which mulberry weed has been collected (specimens plus ad- ditional literature reports) is Louisiana (18), followed by Florida (17), Georgia (11), and Ohio (11). Even though the species was re- ported by Wunderlin (1997) for West Virginia, no specimen could be located to document its occurrence in that state. Based on die herbarium specimens exam- ined, beginning with Louisiana, from which the earliest collection is known (1950), the species appeared in the 1960s in Georgia (1965) and Alabama (1967). In the 1970s it turned up in Florida (1970), Tennessee (1970), Mississippi (1972), Missouri (1972), North Carolina (1973), Texas (1974), Soudi Carolina (1975), Oklahoma (1979), and Ohio (1979). In the 1980s, the species was found in Maryland (1981), Illinois (1982), California (1983), Kentucky (1983), Arkansas (1985), Vir- ginia (1987), and Pennsylvania (1989). In the 1990s specimen were collected in DC (1994), Massachusetts (1994), Indiana (1995), Wash- ington (1995), Utah (1997), and Minnesota (1998). Since 2000, the species has been col- lected in Oregon (2000), Iowa (2000), Michi- gan (2001), Wisconsin (2002), and New York (2002). Spread of Fatoua villosa in North America — Vincent 71 When the distribution ol the specimens is mapped (Figure 2), an interesting pattern emerges with regard to the spread of the spe- cies in the last 50 years. Beginning with the earliest known collection, in New Orleans in 1950, collections gradually radiate out to the southeast, east, northeast, north, northwest, and west. Over the next 50 years, the range of the species gradually increases until the pres- ent-day extent is reached. It cannot be dis- cerned from the available data whether mul- tiple introductions occurred into the United States, or if the species spread from an initial introduction site in New Orleans. The source of the initial introduction is also unknown. Spread of the species does seem to result from horticultural practices. Of the herbarium specimens studied, 50% are from gardens or flower beds, 13% are from nurseries, 12% are from greenhouses, 7% are from lawns, 5% are from potted plants, and 2% are from other horticultural sources (compost piles, mulch, soil piles). Of the remainder, sites from which specimens were collected included old fields, fencerows, cracks and crevices, railroad yards, roadsides, waste places, and dirt or gravel parking lots. Only 6% were from sites away from cultivated areas, where die populations could be considered "naturalized." CONCLUSIONS Fatoua villosa is an increasingly common and widespread weed in the continental Unit- ed States. It is apparently spreading by means of both plants and propagules through the horticultural trade. The species may also be spreading by other means, e.g., import of seeds obtained through seed indices/lists (Anonymous 2004a). While most of the known specimens are from horticultural sites, such as greenhouses, nurseries, and gardens, the spe- cies has spread from these areas into adjacent ones, such as lawns, fencerows, and other In- deral areas. From these habitats, mulberry weed has seemingly spread in some areas to more natural settings, such as forest edges and along streams. Since die species is able to sur- vive and spread in both open and shaded hab- itats, it may spread from horticultural and In- deral settings into more undisturbed sites, es- pecially in open forests, a habitat it occupies in its native range. ACKNOWLEDGMENTS I wish to thank the following individuals for their assistance: N. Harriman (OSH), T G. Lammers (OSH), J. W. Thieret (KNK). I am grateful to the many herbaria cited lor provid- ing access to specimens in their care. LITERATURE CITED Anonymous. 1996. Mulberry weed. Status: exotic in Unit- ed States. Nature Conservancy Priorities for Conser- vation January 1996:13. Anonymous. 2004a. Wuhan Botanical Garden Index Sem- iiinni 2004—2005. littp://www.\vhiob. ac.cn/englisli/web/ indexen.htm (accessed 15 Sep 2004). Anonymous. 2004b. South Carolina Plant Atlas, http:// cricket.biol.sc.edu/lierb/ (accessed 22 Oct 2004). Anonymous. 2004c. Database of Tennessee Vascular Plants, http://tenn.bio.utk.edu/vascular/vascular.html. Anonymous. 2004d. 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Observationes de Urticeis quibus- dam et de Fatoua. Annales Museum Botanicum Lug- duno-Batavum 4:301-307. Nakai, T 1927. Notulae ad plantas Japonicae et Koreanae. XXXIV. Botanical Magazine (Tokyo) 41:501-522. Neal, J. C. 1998. Mulberry weed or hairy crabweed (Fa- toua villosa). Weedlnfo Fact Sheet 03. Horticulture In- formation Leaflet 903. Department of Horticultural Science, College of Agriculture and Life Sciences, North Carolina Cooperative Extension Service, North Carolina State University. Ohwi, J. 1965. Flora of Japan (in English). Smithsonian Institution, Washington, DC. Penny, G. M. 2000. Biology and management of Mulberry weed, Fatoua villosa (Thunb.) Nakai. M.S. Thesis, Hor- ticultural Science. North Carolina State University, Ra- leigh, NC. Penny, G. M., and J. C. Neal. 1999a. Mulberry weed (Fa- toua villosa): a new weed in landscapes and container nursery crops. Proc, So. Weed Sei. Soc. 52:82. Penny, G. M., and J. C. Neal. 1999b. Preemergence con- trol of mulberry weed (Fatoua villosa) in containers. Proceedings of the Southern Nurserymen's Association Research Conference 44:382-^383. Penny, G. M., and J. C. Neal. 2003. Light, temperature, seed burial, and mulch effects on mulberry weed (Fa- toua villosa) seed germination. Weed Tech. 17:213- 218. Randall, J. M. 1997. Weed alert! New invasive weeds in California. California Exotic Pest Plant Council. 1997 Symposium proceedings: 1-6. Randall, R. P. 2002. A global compendium of weeds. R. G. and F. J. Richardson, Victoria, Australia. Reed, C. F. 1977. Economically important foreign weeds: potential problems in the United States. USDA Agri- cultural Handbook No. 498. Washington, DC. Reznicek, A. A. 2001. Mulberry weed (Fatoua villosa) spread as far north as Michigan. Michigan Botanist 40: 73-74. Rohwer, J. G., and C. C. Berg. 1993. Moraceae. Pages 438-453 in K. Kubitzki, J. C. Rohwer, and V Bittrich (eds). The families and genera of vascular plants. Vol. 2. Flowering plants: Dicotyledons: Magnoliid, Hama- melid and Caryophyllid families. Springer- Verlag, Ber- lin, Germany. Sanders, A. C. 1996. Noteworthy collections: California: Fatoua villosa (Thunb.) Nakai (Moraceae). Madrono 43:527. Smith, E. B. 1994. Keys to the flora of Arkansas. Univer- sity of Arkansas Press, Fayetteville, AR. Spread ol Faloua villosa in North America — Vincent SiiihI.II. E. L986. Noteworthy plants from Arkansas. Cas- tanca 51:21 1-215. Swain. L. A., and K. R. Downum. L989. Biologically active coumarins of the Moraceae (fig family). Abstracts of Papers— American Chemical Society L98: AGRO LOO. Swain, L. A., and K. R. Downum. 1990. Liglit-activated toxins in the Moraceae. Biochemical Systematics and Ecology 18:153-156. Taylor, C. E. S., L. K. Magrath, P. Kolley. P. Buck, and S. Carpenter. 1996. Oklahoma vascular plants: additions and distributional comments. Proc. Okla. Acad. Sci. 76: 31-34. Taylor, D. D. 1994. Mulberry weed — a new weedy pest in Kentucky. Kentucky Native Plant Society Newsletter. 9(3):3-l, 10. Taylor, R. J. and C.E. Taylor. 1981. Plants new to Arkan- sas, Oklahoma and Texas. Sida 9:25-28. Thieret, J. W. 1964. Fatoua villosa (Moraceae) in Louisi- ana: new to North America. Sida 1:248. Thomas, R. D„ P. B. Pias Cox, N. A. Dawson, and R. C. Gough. 1980. A checklist of the vascular plants of Morehouse, Richland, and West Carroll parishes of northeast Louisiana. Contributions of the Herbarium of Northeast Louisiana University 1:1-67. Thunberg, C. P. 1784. Flora Japonica. I. G. Miilleriano, Lipsiae. USDA, NRCS. 2004. The PLANTS Database, Version 3.5 (http://plants.usda.gov). National Plant Data Center, Baton Rouge, LA 70874-4490 USA. Vincent, M. A. 1993. Fatoua villosa (Moraceae). mulberry weed, in Ohio. Ohio J. Sci. 93:147-149. Walker, E. H. 1976. Flora of Okinawa and the Southern Ryukyu Islands. Smithsonian Institution Press, Wash- ington, DC. Welsh, S. L„ N. D. Atwood, S. Goodrich, and L. C. Hig- gins. 2003. A Utah flora, 3rd ed. Monte L. Bean Life Science Museum, Brigham Young University, Provo, UT. Wright, R. A. S. 1988. Noteworthy collections: Virginia. Faloua villosa (Thunberg) Nakai. Castanea 53:242. Wu, C.-C, and L.-L. Kuo-Huang. 1997. Calcium crystals in the leaves of some species of Moraceae. Botanical Bulletin of Academia Sinica 38: 97-104. Wunderlin, R. P. 1997. Fatoua. Pages 389-390 in Flora of North America Editorial Committee (eds). Flora of North America. Vol. 3. Magnoliophyta: Magnoliidae and Hamamelidae. Oxford University Press, New York, NY. Wunderlin. R. P.. and B. F Hansen. 2004. Atlas of Florida vascular plants (http://www.plantatlas.usf.edu/). [S. M. Landry and K. N. Campbell (application development), Florida Center for Community Design and Research.] Institute for Systematic Botany, University of South Florida. Tampa, FL. Yama/aki, T. 1082. The seed formation of Fatoua villosa (Moraceae). J. Jap. Bot. 57:350-365. Yatskievych, G, and J. A. Raveill. 2001. Notes on the in- creasing proportion of non-native angiosperms in the Missouri flora, witb reports of three new genera lor the state. Sida 19:701-709. Zhou, /.. and M. G. Gilbert. 2003. Moraceae. Pages 21- 73 iii Z, Wu, I' II. Raven, and I) Hong (eds). Flora "l China. Vol. 5. Ulmaceae through Basellaceae. Science Press, Beijing and Missouri Botanical Garden, St. Lou- is, MO. APPENDIX Selected Specimens Examined (only 1 specimen per county; * = greenhouse weed) ALABAMA: Chambers County, L. Dalrymple s.n. (AUA); Conecuh County, A.R. Diamond 3650 (CA, NLU); Elmore County, E.R. Bums s.n* (AUA); Lee County, J.M. Moffett s.n. (TENN); Madison County. C.T. Bryson J7.529 (USCH); Mobile County. K.E. Rogers 1819 b LeLong (NCU); Pike County, A.R. Diamond 5330 (AUA); Sumter County, R.D. Thomas 100747 b Pitlman (NLU, NY). ARKANSAS: Drew County, E. Sundett 6S34 b Culdin (BRIT, NLU, UAM); Pulaski County, E. b M. Sundett 12049 (NLU, UAM); Union County, R.D. Thomas 107891 et al. (MO, TENN). CALIFORNIA: Alameda County, R.D. Raabe b Stroth- er s.n.* (A); Kern County, Lapp et al. s.n. (CDA); Riv- erside County, A.C. Sanders 15S32 (UCR); San Bernar- dino County, M. Cohen s.n. (CDA); San Diego County. F. McCutcheon s.n. 6- Avery (CDA); Santa Barbara County. K. Cheesman s.n. (CDA). DISTRICT OF COLUMBIA: Washington, S. McNaull 141 b Csiba (MU). FLORIDA: Broward County, M.A. Vincent 10998 b Hickey (MU); Citrus County, E Damian s.n. (GH, NCU); Dade County, /. Popenoe 867 (FTG, NCU); Franklin County, L.C. Anderson 8671 (MO); Hillsborough County. J.M. Kunzer 556 (USF); Leon County. R.K- Godfrey 72357 (GA, MO, NCU, NY, TENN); Monroe County.;. Popenoe 1979 (FTG, NCU); Pasco County, D. Du- Quesnay s.n. (A, CM, FTG, GA, MIN, NCU, NLU, SMU, USF); Sarasota County,/. Popenoe 1979 (USF). GEORGIA: Bibb County, JR. Allison 3514 (GA); Clarke County, W.H. Duncan s.n* (GA, LSU); Columbia County, /. Allison 942 (BH, GA, MO, NCU); Fulton County, R.D. Thomas 101286 et al. (NLU); Glynn County, W.H. Duncan 30555 (GA, MO); Gwinnett County, SB. Jones 22525 (GA, NCU); Jones County. W.H. Duncan 30564 (GA, WIS); Lowndes County. WR. Faircloth 6736 (GA, MO, NCU); Sumter County, R.A. Norris 6710 (GA, NLU); Walker Counh, W.H. Duncan 30647 (MO). ILLINOIS: Champaign County, S.R. Hill 33109 (BRIT, MU. NY, TEX, VT); Jackson County, A.C. Knelling 696S (ILL). INDIANA: Floyd County. Maxwell s.n. (JEF). IOWA: Muscatine County, A.W. Cusick 35602 Ml OSH). KENTUCKY: Adair County, J.W. Thieret 60461 ik\K 74 il of the Kentucky Academy "I Science 65(2) Campbell County, /.W Thieret 57160 (KNK); Jefferson C tv. M Medley 9311-83 (KNK); Madison Count) M.A. Vincent 7163 (KNK. MO, MU). LOUISIANA: Caddo Parish, R.D. Thomas 166976 & Raymond (NLU); Calcasieu Parish, R. Ni'i/land 1143 (LSU); Concordia Parish, R.D. Thomas 24566 (NCU, NLU, TENN); East Baton Rouge Parish, S. Tucker s.n. (LAF, LSU, NLU, TEX); Iberia Parish, R.D. Thomas 19706 et id. (NLU); Jefferson Parish, T. Zebryk 3297 (NLU); Lafayette Parish, J.W. Thieret 16171 (A, LAF, SMU); Lafourche Parish, R.D. Thomas 79963 et al. (BRIT, NCU, NLU); Lincoln Parish, A.W. Boyd 3208 (NLU); Morehouse Parish, R.D. Thomas 56634 b Pias (NLU); Natchitoches Parish, W.C. Holmes 4001 (NLU, NO); Orleans Parish,/. Ewan 23056 (NO, WIS); Ouachita Parish, R.D. Thomas 17629 (GA, ILL, NLU, USF, WTU); Rapides Parish, E. McWilliams M539024 (NLU); St. Charles Parish, G. Montz 5294 (BRIT, LSU, LAF, NLU, NO); St. John Parish, G.N. Montz 5946 (LSU, NLU, NO); St. Tammany Parish, T. Zebnjk 3301 (NLU); Tangipahoa Parish, G.N. Montz 8932 (LSU, NLU, NO). MARYLAND: Baltimore County, C.F. Reed 111095* (MO); Harford County, C.F. Reed 126949* (MO); How- ard County,/. Duke s.n. (NA); Montgomery County, EG. Meyer 22475 (A). MICHIGAN: Jackson County, A.A. Reznicek 11300 (DOV, GH, ILLS, MO, MU, OSH). MINNESOTA: Anoka County, B.A. Addison s.n.* (MIN). MISSISSIPPI: DeSoto County, C.T. Bryson 18056 et al. (NLU, TENN); Forrest County. K.E. Rogers 7902 (NCU); Franklin County, C. Havran 1238 (MISS, MIS- SA); Grenada County, C.T. Bryson (TENN); Lafayette County, M.S. Huneycutt s.n. (MISS); Monroe County, J.R. MacDonald 9258 et al. (DOV); Oktibbeha County, C.T. Bryson 19176 ir Bryson (CM, DOV, MO); Pearl River County, C.T. Bryson 16933 it Sudbrink (MISS); Washing- ton County, C.T. Bryson 15655 (GA, KNK, MO, NLU, TENN, UNLV). MISSOURI: Boone County, P.M. McKenzie 1629 (MO); Butler County, S. Hudson 956 (MO); Cape Girar- deau County, T.E. Brooks s.n. (MO); Cole County, T.E. Smith 3605 (MO); St. Louis County, M.A. Vincent 6443 (MO, MU). NEW YORK: Rensselaer County, N.G. Miller 15565 (GH, MU). NORTH CAROLINA: Brunswick County, M.A. Vin- cent 8681 (BAYLU, C, DAV, ID, ILLS, MU, NCU, OSH, TEX); Cherokee County, E. Lunsford s.n. ir- Morrow (NCU); Dare County, M.A. Vincent 9437 (MU); Durham County, C.F. Reed 116878* MO Iredell County//! Nelson 2809 6 Wnek (NCU); Mecklenburg County, J.F. Matthews s.n. (NCU); Moore County, B.A Sortie 9363 (GH, NCU). OHIO: Athens County, J.W. Thieret 56353* (KNK Butler County. M.A. Vincent .5693 (BIIO. MICH, MO, MU, NY, OS, US, USF); Darke County M.A. Vincent lltltll et al. (MU); Delaware County, AW Cusick 35744 (MU); Franklin County, A.W Cusick 30637 6 Shelton (MU, OS); Hamilton County, M.A. Vincent 7997 (MU, OS); Lawrence County, A.W. Cusick 34753 (CM, MU, NY); Meigs County, A.W. Cusick 35026 (MU); Musking- um County, L.E. Brown 9235* (BRIT); Portage County, A.W. Cusick 32216* (MU); Washington County, A.W. Cusick 30632 6- Ortt (MU, OS). OKLAHOMA: Bryan County, C. Taylor 36151* (OKL); Carter County, A. Buthod 4372 et al. (OKL); Grady Coun- ty, L.K. Magrath s.n. (OKL); McCurtain County, C. Citty s.n. (NLU). OREGON: Benton Count)', R.R. Hahe 5718* (NY). PENNSYLVANIA: Allegheny County, S.A. Thompson 6354* (CM). SOUTH CAROLINA: Florence County, L. Swails s.n. (USCH); Georgetown County, /.B. Nebon 21469 (USCH); Greenville County, N.E. Mullins 75318 (NLU); Lexington County, J.B. Nelson 11950 (USCH); Newberry County, C.N. Horn 4294 (USCH); Pickens County, S.R. Hill 20097 (BRIT. GH, MO, NY. USF); Richland County, J.B. Nelson 5737 (USCH). TENNESSEE: Davidson County, M. Guthrie 609 (TENN); Hamilton County, /.T. Beck 3866 (TENN); Knox County, A.W. Cusick 32241* (MU); Shelby County, A. Evans s.n. (MO, NLU, TENN). TEXAS: Austin County, M.H. Mat/field 1776 et al. (BRIT, MO, TEX); Bexar County, M.A. Vincent 4943 (ILL, MO, MU); Blanco County, R.W. Sanders 5563 (BRIT); Brazoria County, R.J. Fleetwood 11180* (SMU); Collin County, S.R. Hill 4539* (GH); Dallas County, B.L. Lipscomb 3386 (LSU, MU); Gillespie County, R.W. Sand- ers 5339 (BRIT); Harris County, L.E. Brown 8466 (NLU, SMU); Tarrant County, B. Lipscomb 3472 (BRIT). UTAH: Washington County, L. Higgins 19902 (MO, NY). VIRGINIA: Chesterfield County, W.J. Hayden 3553 (NLU, URV); City of Lynchburg, C. Leys s.n. (MO, MU); Fairfax County, T. Darling s.n. (GMUF); Richmond County, R.A.S. Wright 2417 (VPI). WISCONSIN: Winnebago County, M.A. Vincent 10805 6 Lammers* (MU, OSH). List ill Recent Reviewers 75 We gratefully acknowledge the contribution of time and expertise provided !>y the following individuals in reviewing manuscripts submitted for consideration by the Journal. J. Richard Abbott Charles A. Acosta George W. Argus John R. Raird Richard L. Royce David M. Brandenburg Wayne Bresser Jerry H. Caqienter James Duval] Richard L. Josephs Robby Lee [ames O. Luken Joseph I. Orban Mark Pvron Thomas C. Rambo Matt W. Raymond Thomas Sproat David D. Taylor Jerry W. Warner Maiy Kathryn Whitson I kx Vcad Sci 65 2 :76 M 2004. Distributional Records of Selected Kentucky Fishes Michael C. Compton,' David J. Eisenhour,5 Ronald R. Cicerello,3 Lewis E. Kornman,4 Albert Surmont Jr.,' and Ellis I,. Laudermilk3 Kentucky Division of Water, Frankfort, Kentuck) 40601 'Department of Biological and Environmental Sciences, Morehead State University, Morehead, Kentucky 40351 'Kentucky State Nature Preserves Commission. Frankfort. Kentucky 40601 'Kentucky Department of Fish and Wildlife Resources, Northeast Fisheries District, Morehead, Kentucky 403.5 1 ABSTRACT Distributional records for 14 species of fishes are included for Kentucky. Phoxinus oreas is recorded for the first time from Kentucky (Big Sandy River drainage, Pike Co.). Reported for the first time are drainage records for Ichthyomyzon unicuspis and Clinostomus funduloides (Licking River drainage, Batli/Rowan and Morgan cos., respectively); Scaphirhynchus platorynchus (Kentucky Riser drainage, Franklin and Henry/ Owen cos.); Cyprinella gidactura (Green River drainage, Barren Co.); Notropis maculatus (Tennessee River drainage. Graves Co.); and Forbesichthys agassizii (Tradewater River drainage, Caldwell Co. and Crooked Creek system, Crittenden Co.). Range extensions in the upper Cumberland River drainage for Etheostoma crossopterum, E. nigrum, and E. percnurum (Clinton, Pulaski, and McCreary cos., respectively) and for Etheostoma chlorosoma in the Green River drainage (Ohio Co.) are reported. The continued occurrence of the rare fishes Acipenser fulvescens (Ohio River, Lewis Co.), Notropis maculatus (Obion Creek system, Hickman Co.), Noturus exilis (lower Cumberland River drainage, Trigg Co.), and Lota Iota (Ohio River, Bracken, Jefferson, and Livingston cos.) is noted. INTRODUCTION Kentucky has the fourth highest fish diver- sity in the United States, behind Alabama, Georgia, and Tennessee. Clay (1975) reported ca. 200 species occurring in tire state; Burr and Warren (1986), 242 species. During die past 18 years die state total has increased with the resolution of several species complexes (Ceas and Page 1997; Page et al. 1992; Page et al. 2003; Wood et al. 2002). Continued monitoring of the distribution and status of the fish fauna is necessary for resource man- agers in species conservation planning and res- toration efforts. In recent vears, Burr et al. (1990), Warren et al. (1991), Cicerello and Laudermilk (1996), Ryon and Carrico (1998), and Eisenhour and Burr (2000) provided in- formation on the status and distribution of several species. However, collections by uni- versity and state agency personnel, and re- cords of species from sport and commercial fishing infrequently get reported outside of "gray" literature. Herein, the presence of one new species, substantial range extensions of several species, and current status information of some rare species for Kentucky is noted. MATERIALS AND METHODS All records reported were from collections made within die past 10 years by the authors or personnel from the Kentucky Division of Water (KDOW) or die Kentucky Department of Fish and Wildlife Resources (KDFWR), unless noted otherwise. Specimens were col- lected via seining or backpack eleetrofishing, unless noted otherwise. Preserved specimens were deposited at die Illinois Natural Historv Survey (INHS), Morehead State University (MOSU), or Southern Illinois University at Carbondale (SIUC). For specimens not vouchered in museum collections, die authors confirmed identification by personal examina- tion of the specimens or examination of pho- tographs. Common and scientific names for each species follow Nelson et al. (2004). Lengths are reported in standard length (SL) or total length (TL). Species accounts provide museum catalog number, number of speci- mens and their size range (in parentheses), stream name, receiving stream watershed (in parentheses), locality county, and collection date. RESULTS Silver Lamprey Ichthyomyzon unicuspis Hubbs and Trautman MOSU 1588 (1; 111 mm TL), Licking River (Ohio River), in tailwaters below Cave Run Lake dam, Bath/Rowan cos., 23 Mar 2000. 76 Distributional Records — Campion ,.| — \e-«,r°- These radial functions are dien multiplied by die real spherical harmonics to get the complete orbital. The second choice made to reduce run- time is the use of a pseudopotential so that die core electrons do not have to be treated explicitly The pseudopotential is designed to reproduce die wave function and density of the full potential exacdv outside a certain ra- dius from the atom. This calculations uses die pseudopotential developed by Hamman, Schulter, and Chiang (1979). For GaN and InN it was necessary to treat die d electrons Figure 1. Orbitals used in LDA Calculation. The atomic orbital is the dashed line; the solid line is die reduced-range orbital used in this work. The reduced-range orbital is renormalized to produce a neutral atom. They are shown here unrenormalized to compare the shape and range of die reduced-range orbitals to the atomic orbitals. The horizontal axes are in Bohr radii (a„). The vertical axes use units of a,,"1'2 for the s orbital and a^1- for the p orbital. LDA Calculation — Yoder s: ALUMINUM BORON GALLIUM INDIUM NITROGEN ss fourna] ol the Kent i itk\ Academy ol Science 65 2 Table 1. Orbitals used for LDA Calculation. The coefficients and deca) constants for atomic orbitals defin equation (1) are listed below. The a,'s are in units of a,, ' and A,'s are in units of a^ ;. ) i i " .\ a, A. °. ". B i 1 0.2115 0.7700 0.3105 0.7274 2 0.6777 0.2615 0.6230 -0.5936 3 1.9385 -0.9191 1.1540 1.2588 4 .5.4724 0.4757 2.1625 -0.0020 5 15.3507 -0.2239 4.1663 0.7533 N 1 0.5382 -4.3934 0.3917 0,5512 2 0.8377 8.4704 0.8373 0.4601 3 1.3250 -10.2842 1.9694 1.5532 4 2.1410 6.3572 4.7858 2.0949 5 3.5170 -0.9203 11.8171 1.4368 Al i 1 0.2020 -1.2501 0.2184 -0.5284 2 0.4548 1.294.3 0.4720 0.3872 3 0.8910 -0.7715 0.9742 -0.4479 4 1.6709 1.1736 1.9757 0.6019 5 3.0719 -0.9092 3.9768 -0.7207 6 5.5922 0.3445 7.9776 0.7320 7 10.1273 -0.0532 15.9776 -0.4556 Ga i 1 0.1615 -0.7517 0.1916 0.6490 0.2198 -0.0378 2 0,5191 0.2330 0.3250 -1.1594 0.2801 0.1042 3 1.5350 0.6502 0,5170 1.7919 1.3568 2.0812 4 4.3605 -0.3837 0.8012 -1.5096 4.5796 17.8218 5 12.2633 0.1409 1.2234 0.5238 11.1092 35.6777 6 29.1552 -46.3553 In i 1 0.1763 -0.9939 0.1145 0.1847 0.1617 -0.0058 2 0.3810 0.3646 0.2S15 0.0557 0,5291 -0,3409 3 0.7856 0.9248 0,5738 -0.0496 1.4766 -2.7556 4 1.5898 -0.4106 1.0953 -0.1288 3.9508 -6.9840 5 3.1898 0.0475 2.0305 0.1580 10.6424 7.9237 6 3.7118 -0.1109 28.7114 -7.2654 7 6.7351 0.0405 of Ga and In explicitly, since diese states in- teract strongly with the deep 2s electrons in N. So a new pseudo-potential was developed using the same mediods as Hamman et al. to handle these interactions. The new pseudo- potential, with the inclusion of the "semi-core" d-electrons, improved the prediction of die bond lengths from 7.4% error to 3.0% error in zb GaN with similar improvement for InN. The diird choice was to use die Harris Ap- proximation (Harris 1985), which allows a number of different procedures to be imple- mented. The electron density of the crystal is chosen to be die linear combination of the atomic densities. This means that each atom remains neutral during the entire calculation, which, in turn, means die potential is short- ranged. The attractive core term and die re- pulsive Coulomb term cancel one another at large distances from the atom. These two po- tentials are combined and treated as a single potential, which is easily fitted to linear com- 1 .1 ) \ ( lalculation — Yoder S9 Tabic- 2. Equilibrium Crystal Structure of Nitrides using tlir fast I. DA Method. Experimental results Lambrecl .cud Segall, 1994) arc shown in parentheses. The lattice constant, a, is listed in Angstroms. Structural parameters e/a and u arc the commonly defined imilless ratios lor wz. Compound Crystal structure " . onl) u wz onh BN zb 3.653 (3.616) — — wz 2.575 1.649 0.3742 AIN /.I) 4.19S (4.369) — — wz 2.985 (3.111) 1.6(10 11.600) 0.382 (0.385 GaN zb 4.367 (4.501) — — wz 3.087 (3.180) 1.629 (1.624 0.376 (0.375 InN zb t.912 4.97) — — wz 3.473 (3.533) 1.633 (1.6111 0.375 (0.375- biaation of gaussian functions. This allows for rapid, analytic evaluation of matrix elements, since the orbitals are also linear combinations of gaussians. Since this potential is linear in the density these matrix elements can then be stored in large arravs. The matrix element needed at run-time is then simply interpolated from this table. Only die exchange-correlation term resists tabulation and analytic treatment due to the fact diat it is not a linear function of density. To treat this term we fit the exchange-corre- lation potential to a linear combination of gaussian functions in the same manner as die Coulomb-core term. However, unlike the Coulomb term, die fit must be done at run- time. Due to the complicated form of die ex- change, the fit is imperfect but results in a very reasonable approximation of this poten- tial'. Using the local orbitals and the single cal- culation of the matrix elements in the Harris approximation, the calculation has the form of an ab initio tight-binding (TB) calculation. As such, it provides a useful way to conceptualize the interactions taking place in the crystal. The calculation has been used as the foundation of a semi-empirical model for die ground-state molecular structure and quantitative band structure for organic polymers. This model has shown itself to be a \*erv durable and trans- ferable method for calculating band gaps and bond lengths of many different types of con- jugated, organic polymers (Yoder et al. 1999). This was possible because it has all the char- acteristics of a TB calculation but avoids the tedious fitting of a large number ol parameters to experimental results that is found in purely empirical models. This method has also proven itseli to be a good method to produce structural properties of III-Y and Il-YI semiconductors. Bond lengths are generally within 2% of experimen- tal values, and bulk moduli are within 10% (Dickerson 1997; Yoder et al. 1999). These re- sults compare favorably to other slower, more rigorous LDA methods. For the III-N crystals in this work, the re- sults of this method using the orbitals defined b\ Table 1 are shown in Table 2. Also the band structure of the crystal can be extracted from die electron states produced by the calcula- tion. The band structure produced by this cal- culation is qualitatively the same as the band structures that were calculated with a full-po- tential, self-consistent LMTO LDA calculation done bv Lambrecht and Segall (Lambrecht and Segall 1994). This comparison is made in Figure 2 for zb-AlN and wz-GaN. Besults for the other nitrides are similarly consistent with the more rigorous calculations. EMPIRICAL CORRECTION To make this method quantitative we have included small correction terms (much like a TB model) that we can fix to experimental band gaps and lattice parameters. Specifically all that is needed to get the correct band gaps of the material is to shift the cation on-site s- states up to increase the size of the band gap at the experimental lattice parameters. Only in BN does the correction need to be applied to all the on-site B matrix elements, as the B s- and p-states are strongly intertwined for this crystal (Lambrect and Segall 1994). Also, the correction for BN reduces rather than increas- es the size of the band gap as tin- Harris LDA tends to calculate a band ^ ^L --/ 12- 12 i 10- / N <£- 10 - 8- -C-_-^ \ /^- ^v/ 8 - 6- 6 - 4 - 4- 2- 2 -i 0- 0 - -2- -2 - -4- -4 - -b- ■^^ s7" — 7" ^\\ -6 J -8- K ~^= // / \v ^ -8 - -10 - -10- -12- ^ — y -12 - 14- -14 - — -"*"" -1b -1R- -16 - -18 - -20 - yj -22- -22 - X W L r K X M K H K 14 sC\ /* S=L 3 /"T^ 12 15 N; — ' - 10 i \- 8 — 3 - 6 — /~\ \. — 4 \ 1 s 2 — -"\^--^ 5 3 15 ^ST*\ -2 - \ V\ > -4 - \. \\, VI -6 _ ; 1 - -8 - -10 i - -1? 1 -14 \^/ - -IB X W L r K X 12 ^yC&^j 5p-<5 - 10 - --\ i 8 6 — f 3 \ — 3 ^ 4 2 — 3/ — 0 1 4 - -2 _i\ ^r/jy Wk\ -v- -4 "™**V; 1 /- -8 _ i \^_ 3 l"~^ _ 10 - 12 ^ 3 ■ - ^T - 14 1 i - 16 ! "~^^^ 3 - M K M L H K (a) zb-AIN (b) wz-GaN Figure 2. Basic Band Structure of Nitrides, (a) The left column is the band structure of zinc-blend AlN calculated using the method of diis work (top) and calculated using a full-potential, self-consistent LDA method (bottom) (Lam- brect and Segall 1994). (b) The right column is die band structure of wurtzite GaN calculated using the mediod of this work (top) and calculated using a full-potential self-consistent LDA method [4] (bottom). shown in Table 3. Since this correction is only added to states in the valence band, it affects the total energy only indirectly. To produce the measured lattice parameters of the mate- rial a small (~1 eV per bond at equilibrium bond length) exponentially decaying term is added to the total energy. The parameters of this scalar correction are listed in Table 3 as well, where A and a represent the coefficients and decay constants in the equation: R(r) = Ae-"r (2) The final corrected results for these mate- rials, both in die zb and wz crystal structures, LDA Calculati -Yode 91 Tabic 3. Num erica] Parameters used for an Empirical Correction to U)A Calculation. sluit of on-site mntrto elements Element Shill (eV) B -0.403 (p-states) Al 0.879 (s-states) Ga 1 .256 (s-states) In 1 .303 (s-slates) Panimcter Cor scalar i nerg) term defii rd in equation (2). Bond » i \ ' \ , \ B-N 4.0 1 1 1 1 Al-N 4.0 373.35 Gil-N 4.0 398.25 In-N 4.0 87.01 are listed in Table 4. Once we have corrected results for these materials we can apply this correct model to larger systems. We have ap- plied these results to small supercells (64 at- oms for zb, 72 for wz) just large enough to contain all third neighbors in the unit cell. An atom is then removed form the center of the supercell, and the first and second neighbors to the vacant site are allowed to relax. The first neighbors relax toward or away from die va- cant site; the second neighbors relax toward or away from die first neighbors. When diese sets of atoms relax, die band structure is re- calculated. The supercell bandstructures are shown in Figure 3 for zb crystals with a N- vacancy and with then with a cation vacancy. The perfect crystal bandstructures (first col- umn in Figure 3) are shown beside the vacan- cy bandstructures to illustrate how the elec- tron states change. Since removing a nitrogen atom causes an odd number of electrons to be removed, one of die bands shown is half-filled and can be considered the donor or acceptor state. This half-filled bands are indicated with arrows on the figure. In all cases, we see that for the N-vacancy, the half-filled band is in- tertwined with some of the conduction bands. This indicates thai it acts as a shallow donor. Although in the case of AlN several of the low- est conduction bands, including the half-filled band, have separated from the higher hands creating a small gap. Similarly, for the cation vacancy the half-filled band is the highest va- lence band and crosses the other filled bands. This suggests it acts as a shallow acceptor. These results are in good agreement with oth- er experiments and with other calculations. FUTURE WORK We have begun establishing a clear, quan- titative picture ot the pure materials that will allow us to then add a wide range of defects and impurities to the materials and determine defect energies and at least the qualitative be- havior of the donor/acceptor states these changes produce. The range of problems this method could dien be applied to would be very large and could be used in the laboratory to test ideas of dopants, dopant concentra- tions, and other factors to guide their pro- gress. Of particular interest is the p-doping of GaN, where a number of possibilities have been raised. It has been predicted that C on an N-site and Be on a Ga-site were better do- nors dian the Mg dopant that is currently be- ing used (Wang and Chen 2001). However, it has also been shown that Be energetically pre- fers an interstitial location over the Ga-site lo- cation (Van de Walle 1996). With our method, we should be able to confirm or contradict these findings and move toward finding a more efficient way to make p-t\pe GaN. With die large supercells that this method can ad- Table 4. Equilibrium Crystal Structure of Nitrides using the Modified LDA Method. Experimental results are shown in parentheses. The lattice constant, a, is listed in Angstroms. Structural parameters c/a and u are the commonl) defined unitless ratios for wz. Band gap is listed in eV. Coi upon nd Crystal structure ■' c7a (wz link u (wz only) Band gap BN zb 3.612 (3.616) — — 6.1 (6.1) wz 2.552 1.646 0.375 7. SI AlN zb 4.369 (4.369) — — 6.03 wz 3.106 (3.111) 1.601 (1.600) 0.383 (0.385) 6.20 6 1 GaN zb 1.503 (4.5011 — — 3.29 (3.3 wz 3.178 (3.180) 1.6.35 (1.624) 0.375 (0.375) ! I" 3.5 InN zb 4.986 (4.97) — — 1.76 wz 3.524 (3.533) 1.633 (1.611) 0.375 (0.375^ 1.92 (1.9) 92 Journal of the Kentucky Academy oJ Science 65 2 Perfect Crystal Nitrogen Vacancy Cation Vacancy BN AIN ..-. GaN A A A A/ ^ \y \y V InN Figure 3. Supercel] Bandstructure of Nitrides. Band structures produced with semi-empirical calculation for supercells of 64 atoms for the zinc-blend Ill-nitride crystals. The first column is the band structure of the pure crystal. The center column is the supercell wiUr a single nitrogen vacancv and the right column is the supercell with a single cation vacancy. The arrows in the last two columns indicate the positions of the highest occupied (half-filled) bands. I, DA Calculation— Yoder 93 dress, we can even perhaps model co-doping, calculate binding energy of donor-acceptor pairs, and study alloying of the materials in addition to doping. Other potential applications include the in- vestigation of the role of hydrogen in the dop- ing process and modeling the dopant profile across interfaces in heterostructures. The role of hydrogen has already been shown to be a key plaver in Mg doping of GaN as discussed earlier. Oxygen impurities are found to pro- duce "DX states" as they are allowed to relax in the crystal and are found in vvurtzite lattic- es, but not in zinc-blende lattices (Chen and Slier 1995). Again the existence and stability of these states are straight-forward results of this calculation. The dopant profiles across in- terfaces in heterostructures which are neces- sary for almost all device applications and re- quire large supercells, could also be studied. Understanding interfacial segregation is criti- cal to controlling doping in these devices. The potential application of this method is great and will contribute not only to practical device development but will also contribute to the fundamental understanding of basic semi- conductor physics. ACKNOWLEDGMENTS Thanks to Dr. An-Ban Chen of Auburn Uni- versity for useful discussions as well as Ken- tucky NSF EPSCoR for support of this work. LITERATURE CITED Amano. H„ M. Kito, K. Hiramatsu, and I. Akssalri. 19S9. P-type conduction in Mg-doped GaN treated with low- energy electron beam radiation (LEEBI). Japan. J. Appl. Phys. 28:L2112. Chen, A.-B., and A. Slier. 1995. Semiconductor alloys: j >1 in sits and in. iirriak engineering. Plenum Publishing New York. NY, Dickerson, B. K. 1997. Theon ol semiconductor alloys: molecular dynamics of disordered structures, Monte- Carlo simulations of phase diagrams, and efficient AB- initio energ) method. Ph.D. Thesis. Auburn University, Auburn. AL. I lam. inn. D. Pi., M. Seliluler. and C. Chiang. 1979. \orui- conserving pseudopotentials. Phys. Rev. Lett. 43:1494. I [arris, J. 19S5. Simplified method for calculating the en- ergy of weakh interacting fragments. Plivs. Rev. B 31: 1770-1779. Lambrect, W. H. I... and B. Segall. 199-1. Pages 125-127 in J. H. Edgar (ed). Properties of group III nitrides. EM IS Datareviews Series. London. Morkoc. H., S. Strife. G. B. Gao. M. E. Lin. B. Sverdlov. and M. Bums. 1994. Large-band-gap SiC. III-Y nitride, and II-VI ZnSe-based semiconductor device technolo- gies. J. Appl. Phys. 76:1363-1398. Nakamura, S. 1997. Characteristics of room temperarure- C\V operated InGaN multi-quantum-well-structure la- ser diodes. Materials Research Society's Internet J. Ni- tride Semiconductor Res. Vol. 2, Art. 5 and refs. there- in. Nakamura, S., N. Iwasa, M. Senoh. and T. Muki. 1992. Hole compensation mechanism of P-tvpe GaN' films. Japan. J. Appl. Phys. 31:1258. Neugebauer, J.. andC. G. Van de Walle. 1999. Chemical trends for acceptor impurities in GaN. Appl. Phvs. Lett. 85:3003. Sankev, O. K, and D. J. Niklewski. 1989. Ab initio mul- ticenter tight-binding model for molecular-dynamics simulations and other applications in covalent systems. Phys. Rev. B 40:3979-3995. Strite, S.. and H. Morkoc. 1992. GaN. AlN and InN: a review. J. Vacuum Sci. Technol. B 10:1237-1266. van Schilfgaarde. M„ A. Slier, and A.-B. Chen. 1997. The- ory of AlN, GaN, InN and their allovs. J. Crvstal Growth 178:8-31. Wang. H., and A.-B. Chen. 2001. Calculations of acceptor ionization energies in GaN. Phvs Rev. B 63:125212. Yoder, G. B. K. Dickerson, and A.-B. Chen. 1999. Semi- empirical method for calculating structure and band gap of semiconducting polvmers. J. Chem. Phvs. Ill: 10347-10353. J. Ky. Acad. Sei. 65(2):94-103. 20O4. Noteworthy Vascular Plants from Kentucky: A State Record, Range Extensions, and Various Species of Interest J. Richard Abbott Botanv Department. University of Florida, Gainesville, Florida 32611 and Ralph L. Thompson and Rudv A. Gelis Herbarium. Biology Department. Berea College, Berea, Kentucky 40404 ABSTRACT A total of 53 species is presented here, arranged in four groups: (1) a state record, Polygonum densiflonun. (2) range extensions in Kentucky (13 species), (3) various species of interest in the state (34 species, including 12 new populations for species listed and tracked by the Kentucky State Nature Preserves Commission), and (4) cultivated species possibly naturalized in the state (5 species). INTRODUCTION The species reported here primarily repre- sent collections over the last several years based on student-related field research through the Berea College Herbarium (BE- REA). Fourteen new state records invoh~ing BEREA student research were recentiy pub- lished (Abbott et al. 2001). Several Kentucky fioristic projects involving BEREA students have also been published (Thompson et al. 1984, 1996, 2000; Thompson and FitzGerald Jr. 2003; Thompson and Fleming 2004; Thompson and Noe Jr. 2003) and odiers have appeared as abstracts (Abbott and Thompson 1993, 1994; Fleming et al. 1998; Thompson et al. 1995), but reports of most of the notewor- thy species simply have not been formally published. A recent dissertation (Medley 1993) and a recent book (Browne and Athey 1992) both provide fists of the vascular flora of Kentucky, but neither of them includes information from many of the specimens at BEREA. Ongoing research has also yielded additional records since the two checklists. Small regional her- baria such as BEREA are rarely utilized by workers outside the state; thus, dieir holdings remain largely unknown to other investigators. There are several ongoing projects in the state drat may culminate in a fioristic atias, a woody plant flora, and a manual of the flora of Kentucky. We report the following species to make knowledge of their presence available to odier in-state workers and the botanical community at large. Medley (1993) was the primary source for information on die distri- bution of species in Kentucky, but a problem for us is that he occasionally cited a species* presence in a county- based on knowledge of our collections vvidiout referring to our actual specimens. Nonetheless, vvidi very few excep- tions, we only report species here that fill in "gaps" in Medleys report. Browne and Athey (1992) was also referred to, but Medley has most of die same information, typically in greater detail. Any of die species below con- sidered endangered, threatened, rare, or spe- cial concern by die Kentucky State Nature Preserves Commission (KSNPC 2000) are in- dicated after the name bv KSNPC, status, and the year of the listing-report used. Gleason and Cronquist (1991) was used for distribu- tional information outside Kentucky and is die source of our nomenclature. Data on taxa not present in Gleason and Cronquist were found in Radford et al. (1968). Unless odiervvise in- dicated, all specimens are deposited at BE- REA. Fiftv-three species are presented here, placed in four groups: (1) a state record, (2) range extensions in Kentucky (13 species), (3) various species of interest in die state (34 spe- cies, including 12 new populations for species listed and tracked by the Kentucky State Na- ture Preserves Commission), and (4) cultivat- ed species that are possibly naturalized in the state (5 species). 94 Kentucky Plant Records — Abbott, Thompson, anil Gelis 95 KENTUCKY STATE RECORD Polygonum densiflorum Meissner [Polygona- ceae] Native coastal plain species that ranges from New Jersev south to Florida, west to Texas. and interior to southern Missouri (Gleason and Cronquist 1991). It was to be expected in Kentucky (Beal and Thieret 19S6), and it is present on the Mississippi Alluvial Coastal Plain in the contiguous states of Arkansas (Smith 1988), Missouri (Steyermark 1963), and Tennessee (Chester et al. 1997). Based on our examination of Polygonum specimens from Kentucky herbaria. Medley ( 1993) was correct in his assessment that there were not any Kentucky vouchers of P. densiflorum. Our collection is over 500 km east-northeast of any other collections from the above adjacent states. Madison County: Berea College Forest, Red Lick Reservoir No. 2, ca. 2.2 km west of US 421 and KY at Bighill and 0.64 km south off gravel maintenance road; in cattail marsh near the earthern dam; infrequent. 24 Oct 2003; Thompson 03-1208, with J.R. Abbott. RANGE EXTENSIONS Castanea pumila (L.) R Mill. var. pumila [Fa- gaceae] KSNPC Threatened (2000). Johnson (1989) recorded this species as vouchered from eight counties in Kentucky but did not list Madison County. Medley (1993) stated there were five existing popula- tions but that none was known with certainty to exist. Madison County: Anglin Hollow, 2.5 km southeast on Long Branch Road from junction with Red Lick Road (KY 594); occasionally bush-hogged roadside wooded ledge at edge of field: rare, one shrub of several stems. 4 Oct 1993; Abbott 6385, with R.L. Thompson. Crataegus coccinea L. [Rosaceae] Medley (1993) cited only one specimen, from Letcher County without a collector or number. Ross Clark [EKY] first brought this species to our attention bv identifying and an- notating the following sheets (and others at BEREA and EKY). Laurel County: Lilv Surface-mined Area, 0.32 km south of Lilv and 3.2 km east of Kv 25 off Lily-Mcf largue Road; rare, single tree on northwest outslope. 12 [un 1981; Thomp- son 541. with D.D. Taylor. Madison County: Berea College Forest, ca. 3.2 km east of Berea on KY 21: along old pow- erline right-of-way north of road. 22 May 1983; D.D. Taylor 3473. Rockcastle County: John 15. Stephenson Memorial Forest State Nature Preserve, An- glin Falls Ravine; south-southwest trending, dry midslope, rare. 28 Jul 1997; Thompson 97- 161, with C.A. Fleming. Geum laciniatum Murray [Rosaceae] Previously known onlv along the Ohio River and in die coastal plain portion of western Kentucky (Medley 1993). As described in Campbell et al. (1994), the site below was dis- covered bv Rand}' L. Mears in 1993. Laurel Countv: London, 1.2 km south of junction with C.R. 1006 on U.S. 25, on west side of road; swampy bottomland remnant. 19 Jun 1994; Abbott 6999. with R.L. Mears. This site was re-visited on 8 Aug 2002, and it has been partially developed. A large area has been filled in and has several large gravel piles. There is still a small, open, swampy strip adjacent to the nearbv swamp woods, but this species was not relocated. Heracleum maximum Bartr. [= H. lanatum Michx.] [Apiaceae] KSNPC Endangered (2000). A widespread circumboreal species. In Ken- tucky, known widi certainty only from Harlan counts' in the southeast (Medley 1993). Lewis County: Brush Creek Island in the Ohio River; late old-field; rare. 14 Jun 1995: Gelis BC-254. with R.L. Thompson. Lathyrus hirsutus L. [Fabaceae] Native to Europe. In Kentucky known only from seven western counties (Medlev 1993). Madison Count}': Berea College Campus, south of the Alumni Building and Athletic Field, adjacent to Scaffold Cane Road (KY 595); fallow field. 3 Jun 1999; Abbott 12685. Ludwigia hirtella Raf. [Onagraceae] Medley (1993) reported this species from Edmonson and Metcalfe counties, in addition to Pulaski county based on the population re- 96 Kentucky Academy ol Science 65(2) icrcci on 1 nuk l>v poited here, which is voucl collection. Pulaski County: near Woodstock, 2.4 km east on Ocala Road from junction with KY 39, north of Hazeldell Church of Christ; opening in seasonally wet upland woods, succession^ grass-sedge meadow, occasional. 15 Jul 1991; Abbott 1016, with R.L. Thompson. Lycopodium appressum (Chapman) Lloyd & Underw. [Lycopodiaceae] [= Lycopodiella appressa (Chapman) Cran- fill] KSNPC Endangered (2000). Known from Calloway county in the coastal plain portion of western Kentucky. Medley (1993) also cited the population listed here (based on our collection). Pulaski County: near Woodstock, ca. 1.6 km east on Ocala Road from junction with KY 39, north of road along trail; small opening in sea- sonally wet upland woods; rare, only one pop- ulation, fewer than 10 square meters. 5 Jul 1991; Abbott 829, with R.L. Thompson. The population, revisited in 1994, was in stable condition. Ranunculus parviflorus L. [Ranunculaceae] Previously known with certainty only from a few counties in western Kentucky (Medley 1993). Laurel County: off Willie Green Road at Sinking Creek, 0.32 km downstream; fallow field north of creek. 10 May 1994; Abbott 6797, with R.L. Mears. Madison County: Fort Roonesborough State Park; yard in campground; rare. 21 May 1994; Abbott 6911. Ranunculus pusillus Poiret [Ranunculaceae] Frequent in western Kentucky (Medley 1993). Laurel County: London, 0.32 km east of junction with KY 80 on KY 192, dien south of highway; in mowed wet meadow, along small seep. 10 May 1994; Abbott 6801, with R.L. Mears. Ranunculus sardous Crantz [Ranunculaceae] Native to Europe. Frequent in western Kentucky (Medley 1993). These collections show the species to be fairly well established in central and southeastern Kentucky. Rell County: Fonde Surface-mined Dem- onstration Area; mixed hardwoods plantation, outslope; infrequent. 31 May L989; Thompson 89-949. Casey County: adjacent to the intersection of U.S. 127 and 70; in a wet meadow. 1 Jul 1988; B. Hoagland 224 [BEREA; duplicate from EKY; identified as R. pensylvanicus in Hoagland and Jones (1992)]. Knox County: northeast of Corbin, ca. 2 km northwest of U.S. 25 on KY 830; grazed open roadside field; frequent. 25 Jul 1992; Abbott 3344. Laurel County: North Corbin; east of U.S. 25W just north of Whitley County line; upper floodplain of Lynn Camp Creek. 2 May 1999; Abbott 12554. Rockcastle County: east of Disputanta, 0.3 km east on Anglin Fork Road from Ham- monds Fork Road, at junction with Anglin Creek and Clear Creek, wet open field; fre- quent. 9 May 1992; Abbott 2090. Ranunculus sceleratus L. [Ranunculaceae] Frequent along the Ohio River and in west- em Kentucky (Medley 1993). Madison County: Fort Roonesborough State Park; sandy, open, upper beach along river; rare. 10 Jun 1992; Abbott 2519. Urochloa platijphijlla (Munro ex Wright) R. Webster [= Brachiaria platyphylla (Munro ex Griseb.) Nash] [Poaceae] Native southeastern species (Radford et al. 1968). Previously known in Kentucky only from four western counties (Medley 1993). Madison County: Fort Boonesborough State Park; river sand and mudflats; rare. 16 Aug 1992; Abbott 4029, with R.L. Thompson and R.L. Mears. Veronica polita Fries [Scrophulariaceae] Native to Eurasia. Reportedly rare in Campbell and Mason counties (Medley 1993). Medley (1993) reported Veronica agrestis (a very similar species) as frequent "probably throughout die state." We have never seen that species but have seen V. polita regularly enough to suspect diat it is undoubtedly more common than collections indicate and that there may have been some confusion in Med- ley's report. Boone County: soudi of Petersburg, 6.24 km north on KY 20 from junction with KY 18, Kentucky Plant Records — Abbott, Thompson, and Gelis 97 then northwest at junction with Woolper Creek; open field along lake on creek. 13 May 1994; Abboll 6838, with R.F.C. Naczi and R.L. Mears. Garrard County; Camp Nelson Quarry, 2.72 km off KY 1845 from junction of U.S. 27 and 1.6 km west-northwest of Lamber Methodist Church; xeric aggregated gravel floor; infre- quent. 1 Apr 1997; Thompson 97-189, with C.A. Fleming. Jessamine County: waste place along Ken- tucky River, near Brookhii Bridge. 30 Mar 1956; DM. Smith 1261. Madison County: Fort Boonesborough State Park; yards; frequent. 5 Mar 1992; Ab- bott 1322, with R.L. Thompson and G. Dan- deneau. VARIOUS SPECIES OF INTEREST Achyranthes japonica (Miq.) Nakai [Amaran- thaceae] Native to eastern Asia. Previously reported for three counties along Tug Fork in eastern Kentucky. Medlev (1993) also mentioned that the species is probably spreading, as con- finned by this collection and by Vincent and Cusick (1998), who reported it new to Ohio. Lews County: Brush Creek Island in the Ohio River; forested wetland; abundant. 3 Sep 1995; Gelis BC-1077. with R.L. Thompson. Aconitnm uncinatum L. [Ranunculaceae] KSNPC Threatened (2000). Previously reported from five counties from northern to southeastern Kentucky, including Laurel (Medley 1993). This collection repre- sents a new population. Laurel Count)': London, University of Ken- tucky Feltner 4-H Camp; gravelly streambank; rare. 25 Sep 1999; Thompson 99-1060, with E.W.J. FitzGerald Jr. Aegopodiurn podagraria L. [Apiaceae] Native to Eurasia; escaped in northeastern United States. Previously reported only from Jefferson county in Kentucky (Medley 1993). Lewis County: Manchester Island No. 1 in the Ohio River; mature bottomland hard- woods; rare. 8 Jun 1996; Gelis Ml-1211, with C.L. Fleming. Alopecurus pratensis L. [Poaceae] Native lo Eurasia, widespreadlv naturalized in the United Slates. Previously reported as rare in a few counties in northern Kentucky (Medlev 1993). Madison County: Berea College Forest, 1 mi south of Bighill, 175 feet from KY 421 roadside on northwest trending side slope dominated bv serieea lespede/.a. 16 May 2003; Thompson 03-50. with D.B. Poindexter. Cacalia snavcolcns L. [Asteraceae] Known mostly from western Kentucky but also from unverified reports in a few other scattered counties (Medley 1993). Lewis County: Manchester Island No. 1 in the Ohio River; rare along shaded riverbank. 29 Jul 1995; Gelis Ml-767, with R.L. Thomp- son, J.R. Abbott, and A.E. Shupe. Camelina microcarpa Andrz. [Brassicaceae] Native to the Old World. Previously known from a few counties in northern and central Kentucky (Medley 1993). Laurel Counts': London, east of U.S. 25 on J KY 1006 toward Le\i Jackson Wilderness Road State Park, along gravel embankment of railroad tracks. 19 Jun"l994; Abbott 5187, with R.L. Mears. Cardamine impatiens L. [Brassicaceae] Native to Europe and introduced into die coastal states west to Michigan (Gleason and Cronquist 1991). Medley (1993) fisted only Campbell and Jefferson counties in Kentucky-. Lewis County: Manchester Island No. 1 in the Ohio River; late oldfield; frequent. 4 May 1995; Gelis Ml-71, with D. Snell. Carex pedunculata Muhl. [Cyperaceae] When the population below was vouchered, it was only the second known site for this spe- cies in Kentucky (Thompson et al. 2000). Now known in several counties in eastern Kentucky (Medley 1993). Laurel County: Rock Creek Research Nat- ural Area; mixed mesophytic hemlock forest near a small rockhouse recess on conglomer- ate ledge; very infrequent. 21 \pr 1985: Thompson 85-77. Verified by Dr. Robert FC. Naczi. Delaware State University. Chenopodium pumilio R. Br. [Chenopodi- aceael 98 fournal of the Kentucky Academy of Science 65(2) Native to Australia. In Kentucky known pre- viously from Trimble and Fayette counties (Medley 1993). Lewis County: Manchester Island No. 1 in the Ohio River; seasonally flooded and eroded sandy bank, 28 Jul 1995; Abbott 7808, with R.L. Thompson and R.L. Gelis. Chrysosplenium americanum Schwein. [Saxi- fragaeeae] KSNPC Endangered (2000). Rare in southeastern Kentucky (Medley 1993). Already known from Harlan County, but we report a new population. Harlan County: Pine Mountain Settlement School; wooded spring seep into Issacs Creek; infrequent. 26 Mar 1998; Thompson 98-14. Cypripedium calceolus L. var. parviflorum (Salisb.) Fern. [= C. parviflorum Salisb.] [Orchidaceae] KSNPC Threatened (2000). Previously known from a few scattered lo- cations in eastern Kentucky (Medley 1993). Our collection was the basis of Medleys (1993) report from Laurel county. Harlan County: Sheppard Trail, near Pine Mountain Settlement School; on roadside em- bankment. 3 Jun 1997; Thompson 97-1298. Laurel County: Rock Creek Research Nat- ural Area uplands, northeast-facing pine-oak stand; rare. 7 May 1989; Thompson 89-644. Disporum maculatum (Buckl.) Britt. [Lili- aceae] KSNPC Special Concern (2000). Known from several eastern counties in Kentucky, including Bell and Harlan (Medley 1993), but our collections voucher new pop- ulations. Bell County: Fonde Surface-mined Dem- onstration Area; on highwall; infrequent. 21 Apr 1990, Thompson 90-278. Harlan County: Pine Mountain Settlement School; north-trending mesic lower slope along the Split Rock Trail; infrequent to rare. 19 April 1988; Thompson 88-279. Eriophorum virginicum L. [Cyperaceae] Previously known in Kentucky only from Harlan county (Medley 1993). Laurel County: south of Flatwoods and west of Frozen Camp Creek, in powerline right-of- way; wet sandy opening (disturbed drainage channel); several dozen fertile stems seen. 30 Jul 1993; Abbott 5897, with J.J.N. Campbell and S. Walker. Flocrkca proserpinacoides Willd. [Limnantha- ceae] Medley (1993) reported this species as rare near streams along the Ohio River in northern Kentucky, all west of diese collections. Lewis County: Manchester Island No. 2 in the Ohio River; mature bottomland hard- woods; occasional. 4 May 1995: Gelis A/2-75, with D. Snell. Gentiana flavida A. Gray [Gentianaceae] KSNPC Endangered (2001). Previously reported from several scattered counties, some unvouchered or with uncertain identifications (Medley 1993). Madison County: between Brushy Knob and Hacker Smith Mountain, near Lick Fork- Creek; ca. 20 stems at edge of yard and rem- nant cedar glade thicket. 21 Aug 2002; Thompson 02-392, with M. Evans. Geranium dissectum L. [Geraniaceae] Native to Europe. Previously known from northern Kentucky in two counties, with a few other unverified reports (Medley 1993). Madison County: Berea College Campus, south of Kettering Residence Hall and Agri- culture Greenhouses; along edge of cultivated field adjacent to Scaffold Cane Road (KY 595); locally abundant. 23 Apr 1999; Abbott 12458. Hexastylis heterophylla (Ashe) Small [Aristo- lochiaceae] Known previously from Bell and Harlan counties (Medley 1993). Gleason and Cron- quist (1991) treated the species as conspecific under H. virginica. According to Gaddy (1987), this species is distinct from H. virgin- ica. Our plants have predominantly erect calyx lobes mostly 3-4 mm long (characteristics of H. virginica), and the details of the reticula- tion along the internal calyx of some flowers are also reminiscent of H. virginica, with rel- atively low relief and no distinct vertical ridg- es. Thus, we were tempted to follow Gleason and Cronquist. However, some calyx lobes are strongly reflexed and a few are ca. 5 mm long (especially when fresh), and some calices have a pronounced internal reticulation with dis- tinct vertical ridges (characteristics of H. het- Kentucky Plant Records- Aliboll. Timings, son, a in I Gelis 99 erophi/lla). Even the (lowers that are most like //. virgin ica, though, have anther connectives which are exerted beyond the anther (a dis- tinctive //. heterophylla characteristic). Final- ly, Rob Nac/.i, of Delaware State University, who has field experience with many Hexastylis species, looked at our vouchers and then vis- ited the site, concluding that, overall, our pop- ulation is more like H. heterophylla than H. virginica. Laurel County: ca. 100 m west of 1-75 at junction with the Laurel River; a few plants near the river, more common upslope. 9 Apr 1994; Abbott 6532. Liiuiin usitatissimum L. [Linaceae] Known previously from Fulton County in western Kentucky (Medley 1993) and report- edly vouchered from Madison County by Lib- by et al. (1997), although, contrary to the re- port, no specimen was left at BEREA. Pulaski County: northeast of Hazeldell Church of Christ, ca. 1 mile west of junction with Alexander Road on Ocala Road; adven- tive in fallow field [near roadside edge]; rare. 2 Jul 1992; Abbott 2847, with R.L. Thompson. Liparis loeselii (L.) L.C. Richard [Orchida- ceae] KSNPC Threatened (2000). Previously reported from a few counties in eastern Kentucky (Medley 1993). Thompson and MacGregor (1987) reported it from Bell County. Clark County: abandoned limestone quarry, 1.4 km west of junction with KY 1924 on KY 418 and 0.32 km east of Lisletown (Halls on the River), adjacent to Kentucky- River; raised spot in cattail-rush marsh; rare. 5 Aug 1994; Thompson 94-701, with E.W.J. FitzGerald, Jr. Lobelia nuttallii Roemer & Schultes [Cam- panulaceae] KSNPC Threatened (2000). Medley (1993) reported this species from four counties in southeastern Kentucky, in- cluding Laurel and Whitley, based partly on our collections. Campbell et al. (1994) report- ed several other populations in Laurel and Whitley counties, also based partly on our col- lections. Laurel County: Lily Surface-mined Area, 0.32 km south of Lily and 3.2 km east of U.S. 25 off Lilv-McHargue Road; old outslope pond embankment; rare. 28 |un 1 98 1 ; Thomp- son 81-653, with D.I). Taylor. McCreary County: 0.8 km north off U.S. 27 from Scott County (TN) line, and west on Cline Road for 0.48 km; wet meadow portion of field adjacent to mixed hardwoods stand; infrequent. 10 Aug 1993; Thompson 93-465. Whitley County: south of Bark Camp, 0.8 km south of FS. 191 from junction with KY 1 193; in a mowed powerline right-of-way along edge of mixed pine-oak woods; rare. 1 Aug 1993; Thompson 93-447, with J.R. Abbott and A.E. Shupe. Lysimachia vulgaris L. [Primulaceae] Native to Eurasia. Rare in three counties along the Ohio River, west of this collection (Medley 1993). Lewis County: Manchester Island No. 1 in the Ohio River; unconsolidated shoreline; rare. 22 fun 1996; Gelis Ml-1263, with CA. Fleming. Oenothera linifolia Nutt. [Onagraceae] KSNPC Endangered (2000). Rare in four western Kentucky counties and McCreaiy County in southeastern Kentucky (Medley 1993). Pulaski County: near Woodstock, 2.2 km east on Ocala Road from junction with KY 39, NW of Hazedell Church of Christ; in fallow- field; rare. 23 Jul 1992; Abbott 3280, with R.L. Thompson. Papaver (labium L. [Papaveraceae] Native to Europe. Only known from Camp- bell and Kenton counties in northern Ken- tucky (Luken and Thieret 1987). Madison County: near Redhouse, Louis\ille and Nashville Railroad right-of-way, 0.32 km north on KY 388 from junction of KY 3372; infrequent. 17 May 1998; Thompson 98-157. Nicholas County: between gas pipeline and roadside off U.S. 68, 4 km southwest of Ellis- \ille; scattered group of 20 plants. 27 May 1995; Gelis 326, with R.L. Thompson and D. Snell. Platanthera integrilabia (Correll) Luer [Or- chidaceae] KSNPC Threatened (2000). A federal candidate for listing (USFWS 1999). Medley (1993) listed three counties from southern Kentucky. KM) Journal <>l the Kentucky Acadernj ol Science 65(2 Laurel County: Marsh Branch Road (F.S. 774), ca. 0.96 km south of KY 192. then north- east ca. 0.48 km in ravine just north of F.S. 4108; wet swampy streamhead in open woods: one large extended narrow population of hun- dreds of plants. 31 Jul 1993; Abbott 5954. Poa bulbosa L. [Poaceae] Native to Europe. Known from one count)7 in northern Kentucky and three counties in western Kentucky (Medley 1993). Laurel County: London. Le\i Jackson Wil- derness Road State Park; near campground restrooms; forested roadside widi mowed un- derstory. 10 May 1994; Abbott 6776, with R.L. Mears. Madison Countv: Berea, along slope behind buildings at soudieast corner of Chestnut and Boone streets: remnant woodland slope. 20 Apr 1992; Abbott 1911. Ranunculus ficaria L. [Ranunculaceae] Native to Eurasia. Previously known in Ken- tucky from three counties along die Ohio Riv- er, west of diese collections (Medlev 1993). Lewis Countv: Manchester Island No. 1 in the Ohio River; mature bottomland hard- woods; rare. 11 Apr 1995; Gelis Ml-35, with R.L. Thompson. Rliamnus frangula L. [Rhamnaceae] Native to Eurasia. Previously onlv vouch- ered as naturalized in Jefferson and Laurel counties (Medlev 1993). Our site represents a new population. Laurel Countv: London, University of Ken- tucky Feltner 4-H Camp; mesic oak-pine for- est below earthen dam embankment along creek; rare. 16 May 2002; Thompson 02-44, with E.W.J. FitzGerald, Jr. Scirpus fluviattlis (Ton-.) Gray [Cvperaceae] KSNPC Threatened (2000).' Previously listed for two counties in extreme western Kentucky (Medlev 1993), and Jones (1994) reported it from Madison Counts'. Our collection is the easternmost documented for Kentucky. Bell Countv: Fonde Surface-mined Dem- onstration Area; settling pond edge; rare. 12 Aug 1989; Thompson 89-1273. Sherardia aroensis L. [Rubiaceae] Native to western Eurasia and northern Af- rica. Known previously from three scattered counties in Kentucky (Medley 1993). Madison County: Fort Boonesborough State Park; mowed grass-legume field on west side of KY 388; locally abundant. 8 May 1996; Abbott 8565, with R.L. Thompson and B.S. Carlsward. Sicla hermaphrodita (L.) Rusbv. [Malvaceae] KSXPC Special Concern (2000). Medley (1993) reported this species from several other counties along the Ohio River in Kentucky. Lewis Countv: Manchester Island No. 2 in the Ohio River; late oldfield; rare, one eolonv located near island head. 21 Jul 1995; Gelis M2-51 7. Silene ovata Pursh [Carvophvllaceae] KSXPC Threatened (2000). Previously reported from five counties (Medlev 1993), including Bell, which is un- doubtedly based on this voucher. Bell Countv: Log Mountain Surface-mined Demonstration Area; wooded outslope; infre- quent. 26 Jul 1985; Thompson 85-1420, with R.A. Straw" Spiranthes lucida (H. Eaton) Ames [Orchida- ceae] KSNPC Threatened (2000). Medlev (1993) reported this species from seven counties (but some as sight records onlv) in soudiem central to southeastern Ken- tucks-. Clark Counts': abandoned limestone quarry, 1.4 km west of junction with KY 1924 on KY 418 and 0.32 km east of Lisletown, adjacent to Kentucky River; rare, a single eolonv in die Juniperus xeric community at die edge of a small pond. 17 Jun 1994; Thompson 94-426, with J.R. Abbott. Stellaria aquatica (L.) Scop. [= Myosoton aquaticum (L.) Moench] [Carvophvllaceae] Native to Europe. Previously reported as rare on river banks and seeps of five counties along die Ohio River (Medley 1993). Lewis Counts': Brash Creek Island in die Ohio River; unconsolidated shoreline; fre- quent. 27 May 1995; Gelis BC-1S4. with R.L. Thompson. Kentucky Plant R ecorus- -Abbott, Th ompson. and Gelis 101 Vallisneria americana Michx. [Hydrocharita- ceae] Known previously from two counties farther west (than this collection) along the Ohio Riv- er and from a few other scattered counties in Kentucky (Medley 1993). Lewis County: Manchester Island No. 2 in the Ohio River; aquatic lied; infrequent. IT Aug 1995; Gelis M2-860, with H.I, Thomp- son. CULTIVATED SPECIES POSSIBLY NATURALIZED The following species are included as they appeared to us to have been non-cultivated. It is possible, however, that they were planted main- decades ago, although no homesite rem- nants were seen. Some of them were certainly spreading in the immediate area. Nonedieless, it it were possible to know the entire site his- tory, perhaps all but the Euonymus and Li- gustrum could be seen as having spread from cultivation or as persisting and may not truly be naturalized (e.g., Nesom 2000). Akebia quinata (Houtt.) Dene. [Lardizabala- ceae] Native to eastern Asia. Previously- known in Kentucky from Jefferson County (Medlev 1993). Rockcastle Countv: Daniel Boone National Forest, ca. 0.64 km from KY 1004 on a forest service road: roadside pine clear-cut area; rare. 27 Jul 1991; Thompson 91-739, with D.D. Taylor. Euonymus alata (Thunb.) Siebold [Celastra- ceae] Native to eastern Asia. Medlev (1993) re- ported this species as infrequently naturalized in a lew scattered counties in Kentucky. Lewis Countv: Manchester Island No. 1 in the Ohio River;' oldfield; rare. 8 Jun 1996; Cel- ls Ml-1207. with CA. Fleming. Madison County: Berea College Campus, south of the Alumni Building and Athletic Field, along cross-country trail past Brushy Creek; mostly disturbed secondary hardwood forest, but with several scattered, exotic, usu- ally-cultivated plants: apparently spontaneous, dozens of individuals seen along creek. 3 Inn 1999; Abbott 12681. Kerria japonica <\,.> DC. [Rosaceae] Native to China and Japan. Medlev i L993) mentioned ii from cultivation. Gleason and Cronquist (2001) included this species as an occasional escape from cultivation. I Lilian County: I'ine Mountain Settlement School; persisting in mixed mesoplivtie woods from site ol old burned library near Issai s (ieek: rare. 19 Apr HISS; Thompson SS-219. This species was also collected at Pine Moun- tain Settlement School as an apparently culti- vated shrub near a woodland margin far from any buildings (Abbott 8585). This latter collec- tion demonstrates how a planting in an out- of-the-way place could later be seen as possi- bly naturalized, especially when decades have passed and site usage has changed. Madison Countv: Berea College Forest, ca. 3.68 km east of Berea on KY 21 from junction with KY 595; pine-oak woodland strip along north side of road; one multi-stemmed shrub growing at base of, and surrounded by, native trees and shrubs. 23 April 1993; Abbott 4674. Perhaps persisting from cultivation long ago but there is no evidence or local record of an old homesite in the immediate area. Not ap- parently spreading here. This could represent an escape from recent cultivation somewhere nearby. Ligustrum obtusifolium Siebold & Zucc. [Ole- aceae] Native to eastern Asia. In disturbed areas in Favette and Oldham counties (Medlev 1993). Based on several specimens recently annotat- ed at BEREA by Ross Clark (EKY), this spe- cies is much more commonly naturalized than previously reported. Garrard Countv: Camp Nelson Quarry, 2.7 km on KY 1845 from junction with US 27: dry quarry floor near highwall talus area; rare, two fruiting shrubs. 2 Aug 2004: Thompson 04- 1105. Madison Countv: Berea College Campus, south of the Alumni Building and Athletic Field along cross-country trail past Brush) Creek; mostly disturbed secondary hardwood forest, but with several scattered, exotic, usu- ally-cultivated plants; numerous individuals ol various sizes present. 3 |un 1999: Abbott 12682. Rockcastle Countv: |ohn B. Stephenson 102 Journal oi the Kentucky Academy <>i Science 65(2) Memorial Forest State Nature Preserve, An- glin Falls Ravine; northwest-trending lower slope in Pinus-Liriodendron-Quercus com- munity: infrequent. 3 Oct 1997; Thompson 97- 344, with A.N. Allen. Lijcinm barbarian L. [Solanaceae] Native to temperate Eurasia. Documented as sparingly naturalized along a few roadsides and creeks in four Kentucky counties, mostly on limestone-based soils (Medley 1993). Clark Count): roadside ditch along KY 418 ca. 0.16 km from junction widi KY 1924; rare, site discovered by Ross Clark. 26 Sep 2003; Thompson 03-1066. Madison County: Berea College Campus near Agriculture Annex; clustered in cut-back Moms alba. 19 Sep 1985; Thompson 85-1632. LITERATURE CITED Abbott, J. R., and R. L. Thompson. 1993. A relict colony of coastal plain species on the eastern Mississipian Pla- teau in Pulaski County, Kentucky. The 54th Annual Meeting, Association of Southeastern Biologists. Old Dominion Univ., Virginia Beach, VA, 16 April. ASB Bull. 40:149. [Abstract]. Abbott, J. R., and R. L. Thompson. 1994. A floristic survev of Fort Boonesborough State Park, Madison County, Kentucky. The 55th Annual Meeting, Association Southeastern Biologists. Central Florida Univ., Orlando, FL, 14 April. ASB Bull. 41:91. [Abstract]. Abbott, J. R.. R. L. Thompson, and R. L. Gelis. 2001. Vascular plants new to Kentuckv. Sida 19:1199-1202. Beal. E. O., and J. W. Thieret. 19S6. Aquatic and wetland plants of Kentucky. Kentucky State Nature Preserves Comm. Sci. Techn. Ser. 5. Browne, E. T, Jr., and R. Athey. 1992. Vascular plants of Kentucky: an annotated checklist. Univ. Press of Ken- tucky, Lexington, KY. Campbell, J. J. N., J. R. Abbott, R. R. Cicerello, J. D. Riser, J. H. MacGregor, and J. G. Pahs. 1994. Coop- erative inventory of endangered, threatened, sensitive and rare species, Daniel Boone National Forest: Lon- don Ranger District. Kentucky State Nature Preserves Commission, Frankfort, KY. Chester, E. W., B. E. Wofford, and R. Krai. 1997. Atlas of Tennessee vascular plants, Vol. 2. Angiosperms: di- cots, Misc. Publ. 13, The Center for Field Biology, Aus- tin Peay State Univ., Clarksville, TN. Fleming, C. A., R. L. Thompson, and R. A. Gelis. 1998. Wetland and aquatic vascular flora of five reservoirs in the Berea College Forest, Madison and Jackson Coun- ties, Kentuckv. The 59th Annual Meeting, Association of Southeastern Biologists. Northeast Louisiana Univ., Monroe, April 16. ASB Bull. 45:91. [Abstract]. Gaddy, L. L. 1987. A review of the taxonomy and bioge- ograph) ol Hexastylis (Aristolochiaceae I astanea 52: 186-196. Gleason, H. A., and A. Cronquist. 1991. Manual nl\ os- cular plants of northeastern United States and adjacent Canada. 2nd ed. New York Botanical Garden, Bronx, NY. Hoagland, B. W., and B. L. Jones. 1992. Wetland and riparian flora of the Upper Green River Basin, south- central Kentucky. Trans. Kentuckv Acad. Sci. 53:141- 153. Johnson, G. P. 1989. Castanea and Fagus (Fagaceae) in Kentucky. Trans. Kentucky Acad. Sci. 50:75-78. Jones, R. L. 1994. New localities for rare or infrequent vascular plants of Kentuckv. Trans. Kentuckv Acad. Sci. 55:139-141. [KSNPC] Kentuckv State Nature Preserves Commission. 2000. Rare and extirpated plants and animals of Ken- tucky. J. Kentucky Acad. Sci. 61:11.5-132. Libbv. G W., R. L. Mears, and C. T Bloom. 1997. Note- worthy vascular plant discoveries from Kentucky. Trans. Kentucky Acad. Sci. 58:74-79. Luken, J. O., and J. W. Thieret. 1987. Linum grandiflorum (Linaceae), Papaver dubium (Papaveraceae), amd Sal- via pratensis (Labiatae): additions to the Kentucky flora. Trans. Kentucky Acad. Sci. 48:26. Medley, M. E., R. Cranfill, L. R. Phillipe, and R. Hannan. 1980. Sundews in Kentucky and notes on their only ex- isting habitat in die state. Unpublished abstract pre- sented at the 66th annual meeting of the Kentucky Academy of Sciences, on file with the Kentuckv- State Nature Preserves Commission, Frankfort, KY. Medley, M. E. 1993. An annotated catalog of die known or reported vascular flora of Kentucky. Ph.D. Disser- tation. Univ. Louisville, Louisville, KY. Nesom, G L. 2000. Which non-native plants are included in floristic accounts? Sida 19:189-193. Radford, A. E., H. E. Ahles, and C. R. Bell. 1968. Manual of the vascular flora of the Carolinas. Univ. North Car- olina Press, Chapel Hill, NC. Smidi, E. B. 1988. An adas and annotated list of the vas- cular plants of Arkansas. 2nd ed. Privately published by Edwin B. Smidi, Univ. Arkansas Bookstore, Fayette- \ille. AR. Steyermark, J. A. 1963. Flora of Missouri. Iowa State Univ. Press, Ames, LA. Thompson, R. L., J. R. Abbott, and A. E. Shupe. 1995. Vegetation and vascular flora of an abandoned lime- stone quarry, Clark County Kentucky. The 56di Annual Meeting, Association of Southeastern Biologists. Univ. Tennessee, Knoxville, 23 April. ASB Bull. 42:85. [Ab- stract]. Thompson, R. L., and E. W. J. FitzGerald Jr. 2003. Vas- cular flora of Feltner Lake, Laurel County, Kentucky. J. Kentucky Acad. Sci. 64:75-92. Thompson, R. L., and C. A. Fleming. 2004. Vascular flora and plant communities of the John B. Stephenson Me- morial Forest State Nature Preserve (Anglin Falls Ra- Kentucky Plant Records — Abbott, Thompson, and Gelis L03 vine), Rockcastle County, Kentucky. Castanea 69:125- 138. Thompson. R. [,.. R. L. Jones, J. It. Abbott, and W. N. Denton. 2000. Botanical Survey of Hock Creek Re- search Natural Ana. Kentucky. U.S. I). A. Foresl Ser- vice, Northeastern Research Station. General Technical Report NE-272. Newtown Square, PA. Thompson, R. L., ami J. R. MacGregor. 1986. Liparisloe- sclii (Orchidaceae) documented in Kentucky. Trans. Kentucky Acad. Sei. 47:138-139. Thompson, R. L„ and F. D. Noe Jr. 2003. American mis- tletoe (Phoradendron leucarpum, Viscaceae) in Rock- castle County, Kentucky. J. Kentucky Acad. Sei. 64:29- 35. Thompson, R. L., \V. G. Vogel, and D. D. Taylor. 1984. Vegetation and flora of a coal surface-mined area in Laurel County. Kentucky. Castanea 49:1 1 1-126. Tl pson, 11. I... G. L. Wade, and R, A Straw. 1996. Natural anil planted flora ol l.og Mountain Surface- mined Demonstration Area, Hell County Kentucky. Pages 484-503 in Successes and failures: applying re- search results to insure reclamation success. Proceed- ings. 13th Annual National Meeting, American Society of Surface Mining and Reclamation (ASSMR), May 18- 23, Univ. Tennessee. Knoxville, TN. [USFWS] United States Fish and Wildlife Service. 1999. Endangered and threatened wildlife and plants, review of plant and animal taxa that are candidates or proposed for listing as endangered or threatened. Fed. Hrgistei 64:57533-575 17 Vincent, M. A., and A. W. Cusick. 1998. New records of alien species in the Ohio vascular flora. Ohio J. Sei. 98: 10-17. J. Ky. Acad. Sd. 65(2):104-107. 2004. A PCR-Based F, Hybrid Screen Using the Beta-aetin Genes from the Sunfishes Lepomis cyanellus and L. macrochirus (Centrarehidae) David K. Peyton Department of Biological and Environmental Sciences, Morehead State University, Morehead. Kentuck) 40351 ABSTRACT Hybrids between closely related fish species are widely propagated in aquaculture and known to occur in many instances under natural conditions. The resulting hybrids often do not have characters that distinguish them definitively from the multiple possible parent species, making field identification difficult or impossible. This can be particularly difficult with juvenile specimens and adults who are not displaying breeding color- ation. Lepomis species are capable of producing many hybrids, and L. macrochirus X L. cyanellus hybrids are produced commercially and released into private and public waters. This hybrid, easily distinguishable by morphology from its parent species, provided a good model to test a molecular identification tool. In this study I describe a method to design an unambiguous polymerase chain reaction (PCR) screen for hybrids that can be performed on living or dead specimens, requires minimal and non-invasive tissue sampling, is rapid and inexpensive, and can be adapted for any species. INTRODUCTION The problem of correctly identifying hybrid offspring in closely related fish species is com- pounded by the complex genetic interactions that may occur between the chromosomal contributions of the parent species. Morpho- logical traits can often identify candidate pa- rental species but confirmation is seldom pos- sible. Moreover, F! hybrid offspring may vary from each other due to crossing over in pa- rental gametes, and the outcome may also dif- fer between crosses if the gender of the par- ticular species is varied (i.e., male Lepomis cy- anellus X female L. macrochirus versus fe- male L. cyanellus X male L. macrochirus) (Childers 1967). Even non-hybrid juveniles of some species can be difficult to identify cor- rectly without tedious scale counts and dissec- tion. Application of a molecular test with un- ambiguous results provides a definitive medi- od to ascertain correctly the identity of a fish as well as to determine whether or not the specimen is a hybrid. Common molecular techniques for fish identification are allozymes, microsatellite markers, or other DNA profiling strategies such as restriction fragment length polymor- phisms (RFLPs) (Avise and Saunders 1984; Neff 2001). While these are effective and re- liable in most circumstances, characterizing polymorphisms in the genome can be time- consuming, and evaluating the mediods for statistical reliability can be overwhelming and costly. With relatively inexpensive reagents for polymerase chain reaction (PCR) and increas- ing knowledge of fish genomics, more precise strategies are possible. An alternative approach, presented here, is to look for intronic polymorphisms in a con- served gene diat can distinguish species that are otherwise nearly identical at die genetic- level. This approach has several advantages. First, consensus primers to exonic sequences can be used to isolate the gene from multiple species for sequence analysis. Second, poly- morphisms can be chosen that are not hyper- variable in nature. Therefore, these polymor- phisms are likely to be found universally with- in a single species with few exceptions. Third, because introns tend to span large distances, it may be possible to design primer sets that can screen for multiple species with multiple possible PCR product lengths in a single re- action. Lastly, because the length will likely be much longer than a typical microsatellite, anal- ysis can be performed on simple agarose gels instead of high resolution polyacrylamide gels. The gene chosen in this study is the cyto- plasmic beta-actin gene, a member of the ac- tin gene family. This gene is highly conserved across kingdoms, has little or no variation at the amino acid level, and is highly conserved at the nucleotide level in teleosts. There are at least six, and possibly as many as nine, actin family members in teleosts (Venkatesh et al. 1996). Beta-actin can be distinguished from 104 PCR-Based F, Hybrid Screen— Peyton 105 other actins by the signature amino acids at each terminus of the coding sequence. Within the heta-actin gene, the introns are valuable phylogenetic markers (Lee and Gye 2001 ). In this study, primers were designed to hybridize to unique regions in the first intron ol the beta-actin gene in two sunfish (family Cen- trarchidae), creating different products in each species and providing a rapid technique for identification of hybrids. The application of this approach to other species and other fam- ilies is discussed. MATERIALS AND METHODS Specimens Lepomis cyanellus and L. macrochirus spec- imens were collected from South Elkhorn Creek in Woodford Count}', Kentucky, and from Town Branch in Fayette County, Ken- tucky. Lepomis cyanellus X L. macrochirus hybrid specimens were purchased from the Jones Fish Hatchery (Cincinnati, OH). Isolation of Genomic DNA Small fin clip biopsies (5 mm2) from eight fish were incubated in 250 (xl lvsis buffer (100 raM Tris, 5 mM EDTA, 0.2% SDS, 200 mM NaCl), and 5 julI proteinase K (from a 100 |xg/ ml stock solution) at 52°C for 2 hours. Sam- ples were vortexed and centrifuged in a mi- crofuge for 8 minutes at top speed. The su- pernatant was then transferred to a fresh tube containing 400 (jlI isopropanol. The tube was inverted 10 times and centrifuged for 1 mi- nute at top speed, and the supernatant was removed and discarded. The pellet was washed with 95% ethanol, dried briefly, then resuspended in water. Cloning of Beta-actin Genes Beta-actin whole gene primers were de- signed to hybridize with the first 9 and the last 9 codons of the cytoplasmic beta-actin gene based on conserved sequences in available da- tabase entries. Whole gene primers were as follows: Forward: 5' atg gat gat gaa ate gec gca ctg gtt 3'; Reverse: 5' tta gaa gca ttt acg gtg gac gat gga 3'. Cycle parameters were: 95°G for 15 minutes, followed by 30 cvcles of (95°C for 30 sec, 47°C for 30 sec, 72°C for 1 mi- nute). The genomic fragment produced from each fish was ca. 2 kb and was cloned into the pGEM-T Easy vector (Promega). Candidate clones were sequenced and compared to known beta-actin genes for verification, and die exons were determined using consensus splice sites and the \iiiualK invariant lirtu-ac- t i 1 1 amino acid sequence. Amplification ol Polymorphic Regions The polymerase chain reaction was carried out with each sample using the following amounts: 5 |jl1 genomic DNA (from a 10 ng/ (jlI stock), 1.5 u.1 of each primer (from a 10 ng/ u.1 stock), 12.5 uJ of 2X Thermoscript PCR master mix (Marsh Bioproducts), and water up to 25 |jl1. The Exon 1 Forward Primer (EXIF) sequence is: 5' tgg ttg ttg aca acg gat ccg gta tgt gca 3'. The L. macrochirus reverse primer (MRP) is 5' tta aaa ggt aaa gat ctt gac tac atg tac g 3'. The L. cyanellus reverse prim- er (CRP) is 5' tgg tta gac etc att aga tgt cag cat atg 3'. The cycle parameters lor the prim- ers used in this study were as follows: 95°C for 15 minutes, followed by 30 cycles of (95°C for 30 sec, 47°C for 30 sec, and 72°C for 1 mi- nute). Samples were electrophoresed on a 1.5% agarose gel and visualized using ethi- diuni bromide staining. RESULTS Exon 1 and intron 1 of the beta-actin genes from L. cyanellus and L. macrochirus are shown in Figure 1. The first 27 nucleotides of the gene were included in the synthesized primers used to isolate the gene and as such do not necessarily represent die true genomic sequence and are not included here in calcu- lation of identity. All data discussed is based on codons 10-366 out of die 375 total codons. The total intronic sequence between the start and stop codons is 49 nucleotides longer in L. cyanellus than L. macrochirus, but it is nearly identical otherwise (Figure 2). The majority of the differences observed reflect gaps or inser- tions and occur within the first intron. The se- quence identity for the total sequence is 94.6%. The presumed amino acid sequence has over 95% identity with published protein sequences from Oncorhynchus mykiss (John- son 2002) and Cyprinus carpio (Liu et al. 1990). Variation in the coding regions between L. macrochirus and L. cyanellus is confined to the third position of the codon in most in- stances, as expected (data not shown). Primers were designed to exploit the differ- 106 journal of the Kentucky Academy ol Science 65(2) Exon 1 forward primer - L.m. ATGGATGATGAAATCGCCGCACrGG7TG7TGACAACGGA TCCCCTATCT L.c. ATGGATGATGAAATTGCCGCACTGG rrGTrGACAACGGArCCGGrATGr GCAAAGCCGGTTTCGCCGGAGACGACGCCCCTCGTGCTGTCTTCCCCTCCATCG GCAAAGCCGGTTTCGCCGGAGACGACGCCCCTCGTGCTGTCTTCCCCTCCATCG TTGGTCGCCCCAGGCATCAGgtgagtgattgatcgccagcacaataaagccaca TTGGTCGCCCCAGGCATCAGgtgagtgattgatcgccagcacaataaagccaca ccgtttttta tggattttaaaacacatttactatcc ctggtttttaataagaacttgctgattatggattttaatacacatttactgacc taattacactcctaagcaattaaaatta ttcctgaatttcttgattgtta taattacactcctaaacaattaaaattaattattcctgaatttcttgattgtta aatgaaaattgetttget aatgaggtctaaccactaagc g c t g a a a a 1 1 g c 1 1 1 g c t ca_tatg_ctg_aca_tctaa_cg_ag_g_tctaacca c t a a g c * — L. cyanellus reverse primer aacatttacatgcgcaacctgattaaataagtactatattatgggaaatattcc aacatttacatgcacaacctaattaaa tactgcattataggaaatattcc ^ L. macrochirus reverse primer ctcattgtatt tgacacacg tacatqtaqtcaaqa tct ttaccttttaat cacatggtattgccgagacacaaatacatgtagtcaagatcttaaccttt-aat taggaactgctacatatccatgttttgttttt aacaagtttgtcttg taggaactgctacatagcaatgttctctttttttcttttaacaagtttgtattg tttgtcatgtcctgttcagGGAGTGATGGTGGGTATGGGCCAGAAGGACAGCTA tttgtCatgtcttgttcagGGAGTGATGGTGGGTATGGGCCAGAAGGACAGCTA Figure 1 . The genomic sequence of the beta-actin gene in Lepomis macrochirus and L. cyanellus. The exonic se- quence is in capital letters, intron 1 is in lowercase letters. All of exon 1 and the beginning of exon 2 are shown. The locations of primers are italicized. Gaps are indicated by dashes in the sequence. ences observed in the first intron, particularly where gaps were present. PCR amplification with the primers indicated in Figure 1 pro- duced species-specific products when assayed by gel electrophoresis. Figure 3a shows the re- sults when the Exon 1 Forward Primer (EXIF) was paired with the L. macrochirus Reverse Primer (MRP). There are 355 base- pair long bands present with L. macrochirus DNA (lanes 2—4), absent with L. cyanellus 1234 56789 L. macrochirus 429 /\ /\ \ 123 1 330 | » | I 439 1 111 [ 182 [ 1 09 | 144 | I L. cyanellus II III rv V 1 123 | 373 | 239 I 439 III ] 111 | 182 J 108 | 144 | I II IV V Figure 2. Gene structure of the beta-actin gene in Le- pomis macrochirus and L. cyanellus. The nucleotide lengths of exons I through V are indicated in boxes and the lengths of introns are indicated between the boxes. The gene contains four introns between the start and stop codons. 5' and 3' untranslated regions are not included here. Figure 3. PCR products are species-specific. Panel 3a shows the results when the "Exon 1 Forward Primer" is paired with the Lepomis macrochirus Reverse Primer (MRP). The expected product is 355 basepairs long. Bands are only present with L. macrochirus DNA (lanes 2-4) and hybrid DNA (lanes 8-9). Panel 3b shows the results when EXIF is paired with the the L. cyanellus Reverse Primer (CRP). The expected product is 282 ba- sepairs long. Bands are only present with L. cyanellus DNA (lanes 5-7) and hybrid DNA (lanes 8-9). Panel 3c shows the products of a reaction using EXIF, MRP and CRP. Both species exhibit their expected size bands (lanes 2-4 and 5-7), and the hybrid DNA produces both 282 and 355 basepair products. DNA (lanes 5-7) and present with genomic DNA from the hybrid specimens (lanes 8-9). Figure 3b shows the results when EXIF was paired with the the L. cyanellus Reverse Prim- er (CRP). There are 282 basepair long bands absent with L. macrochirus DNA (lanes 2-4), present with L. cyanellus DNA (lanes 5-7) and present with genomic DNA from the hy- brid specimens (lanes 8-9). The products of a reaction using EXIF and both reverse primers simultaneously (MRP and CRP) are shown in Figure 3c. In this sce- nario, each single species is limited to produc- PCR-Based F, Hybrid Screen— Peyton 107 ing the specific hand corresponding to its chromosomal sequences, hut the hybrid sam- ples allow amplification of both the 282 and 355 hasepair products. It may be pointed out that a potential technical limitation is that competition for the common primer in each reaction (in this case, the forward primer) may cause unequal intensity in the resulting bands when both templates are present, as in the hy- brid. Most likely this could be overcome by tedious selection and optimization of primer length and GC content, but the unequal in- tensity does not interfere with the interpreta- tion and is considered unimportant in the final result. The fact that both bands are present only if both parental versions of beta-actin are present is of primary interest here. This limi- tation was not a factor in this primer combi- nation, but to normalize intensity between sin- gle species genomic DNA samples and hybrid DN A samples (which contain half of the tem- plate amount for each product), 30% more hy- brid product was loaded into the gel for anal- ysis. DISCUSSION In this study, I describe a technique for de- veloping a rapid and unambiguous test for identification of Fl hybrid fish. This technique is not suitable for determining introgression, gene flow, or F2 crosses. Instead, it should be used as a simple identification tool and can be rapidly adapted to any species. For example, in 10 days or less the sequence data for the beta-actin gene can be acquired from a fin bi- opsy, and species-specific primers can be syn- thesized. Even though the intronic primers used here, by design, have limited application outside of L. cyanellus and L. macrochirus, the consensus primers for the start and stop codons have potential for use in any species, and could be even more adaptable if the wob- ble codon positions were made degenerate in the primer design. Preliminary data indicate that these primers will not be restricted to the centrarchids. At this time, I have already iso- lated the beta-actin gene from five esocids us- ing these primers and the techniques de- scribed herein. Therefore, there is little doubt that this technique could have broad applica- tion. This is also the first description of a Lepom- is beta-actin genomic sequence. The predicted gene structure appears In have conserved exon lengths with other teleosts as distantly related as Takifugu rubripes (Venkatesh et al. 199fi) and Esox masquinongy (author's observation). The amino acid sequence is highly conserved, and it is noteworthy to mention that beta-actin is a widely used phvlogenetic marker that could have potential lor characterizing the re- lationships between the Centrarchids as well as other families (Baldauf et al. 2000; Bhatta- charva and Weber 1997; Coodson and Hawse 2002). The implications of this will be dis- cussed elsewhere when a complete collection of Lepomis sequences is obtained. ACKNOWLEDGMENTS I thank Ben Brammell for his assistance in the collection of specimens. This work was supported by a grant from the Institute for Regional Analysis and Public Policy at More- head State University. LITERATURE CITED Avise, J. C, and N. C. Saunders. 1984. Hybridization and introgression among species of sunfish (Lepomis): anal- ysis by mitocbondrial DNA and allozyme markers. Ge- netics 108:237-255. Baldauf, S. L., A. J. Roger, I. Wenk-Siefert, and W. F. Doolittlc. 2000. A kingdom-level phylogeny of eukary- otes based on combined protein data. Science 290:972- 977. Bhatlaeharya, D., and K. Weber. 1997. The actin gene of the glaucocystophyte Cyanophora paradoxal analysis of the coding region and introns, and an actin phylogeny of eukaryotes. Curr. Genet. 31:439-446. Childers, W. F. 1967. Hybridization ot four species of sun- fishes (Centrarchidae). Illinois Nat. Hist. Surv. Bull. 92: 159-214. Goodson, H. V., and W. F. Hawse. 2002. Molecular evo- lution of the actin family. J. Cell Sci. 115:2619-2622. [ohnson, M. C. 2002. Direct submission of Oncorhynais mykiss sequence to GenBank. Accession no. AJ43815S. Lee, J. S., and M. C. Gye. 2001. Use of beta-actin gene nitron 2 as a phylogenetic marker in fish taxonomy DNA Sequence 12:71-76. Liu, Z. J., Z. Y. Zhu, K. Roberg, A. Faras, K. Guise, A. R. Kapuscinski, and P.B. Hackett. 1990. Isolation and char- acterization of beta-actin gene of carp (Cyprians car- pio). DNA Sequence 1:125-136. Neff, B. D. 2001. Genetic paternity analysis and breeding success in bluegill sunfish (Lepomis macrochirus). J. Hered. 92:111-119. Venkatesh, B., B. H. Tay, G. Elgar. and S. Brenner. 1996. Isolation, characterization and evolution ol nine puller- fish (Fugii nibripes) actin genes, f. Molecular Biol. 259: 655-665. J. Ky. Acad. Sci. 65(2): 108-1 15. 2004. Growth of Stygobitie (Orconectes australis packardi) and Epigean (Orconectes cristavarius) Crayfishes Maintained in Laboratory Conditions Ann-Simone Cooper and Robin L. Cooper' Department of Biology, University of Kentucky, Lexington, Kentucky 40506-0225 ABSTRACT This study reports on maintenance and growth of the cave crayfish, Orconectes australis packardi, and the epigean crayfish, Orconectes cristavarius, within laboratory conditions for 1 and 2 years. The O. a. packardi survived well compared to the O, cristavarius in captivity. The poor survival of the epigean species was probably due to unsuitable conditions. The epigean as well as the cave crayfish molted and grew in captivity, but without any significant difference in molt frequency between species. In the first year, total body length was obtained to assay growth, whereas in the second year the more accurate measure of post- orbital carapace length was used. The ability of O. a. packardi to adjust to captivity is likely due to their lower metabolic rate and ability to handle hypoxic stress better dian epigean species. INTRODUCTION The growth and maintenance of cave cray- fishes in laboratory conditions is not well doc- umented but advantageous to know for several reasons. Although field studies may inform us of those aspects of growth that occur in na- ture, they are complicated by many variables that cannot be controlled. For instance, with respect to crustaceans as well as mammals, pe- riodic climate changes over a yearly cycle or over several years can bias data obtained dur- ing a brief period of time. This is also relevant for cave crustaceans that are influenced by surface streams varying seasonally in temper- ature. Resources, such as food and shelter, also may impact one subset of the population but not another (e.g., location dependent), as is known to occur in sand crabs (Siegel and Wenner 1985). Standardization of such vari- ables in controlled laboratory studies allows them to be assessed and tested for integration in field studies. In addition, knowledge of how well stygobitie (i.e., aquatic cave obligate) an- imals survive in a holding facility is of use in case of a need to temporarily circumvent spe- cies eradication by acute environmental im- pacts. Such disturbances occur with land de- velopment, producing a high sedimentation and anthropogenic pollutants known to be le- thal to crustaceans in general (Fingerman 1985). Growth measurements of stygobitie cray- 1 To whom correspondence should be addressed. fishes in the field have proven to be a difficult task, and this is likely the reason for the scar- city of quantitative data. Problems of marking and recapture of cave crayfish species over the molt cycle have been addressed by Cooper (1975), Cooper and Cooper (1976), Hobbs III (1978), and Weingartner (1977). Using various marking approaches, individuals have been re- captured and identified for 5 years in one study (Cooper 1975), 3.5 years by Weingartner (1977), and 2 years in another study (Hobbs III 1973). Recapture studies of troglophilic crayfish (Cambanis laevis Faxon) over a year and ones maintained in jars held within sur- face and cave streams were conducted by Weingartner (1977). Field monitoring of crustacean develop- ment in a variety of karsts with wide-ranging dynamics is necessary since water tempera- ture, environmental space, and food resources are not constant within all karst systems. Sur- face stream runoff from summer to winter al- ters water temperature in caves. For example, karst waters in an Indiana cave varied from 11.6 to 8.0°C after a rain on a snow-laden sur- face (Poulson 1964). In the second longest cave in Kentucky (Coral Cave, 38.46 km of passages) and die third longest (Sloans Valley Cave, 37.70 km of passages), both in Pulaski County, stygobitie crayfish (Orconectes aus- tralis packardii Rhoades, 1944), occur within a short distance of a karst window (30 m in Coral Cave) and are exposed to fluctuating wa- ter temperatures. In contrast, the longest cave 108 Laboratory Growth <>l Cave and Surface Crayfishes — Coop (I Cot per KM in Kentucky (Mammoth Cave, Edmonson County, 580 km ol passages) contains deep bodies of water with little variation in temper- ature as well as regions more directly influ- enced hv surface stream temperatures (Barr and Kuelme 1971; Packard INNS; Poulson 1992). Since water temperature is the major environmental trigger for inducing molting in cave crayfishes (Jegla 1966, 1969), one would expect crayfish populations within caves to have varied molt cycles. Thus, growth rates of crayfishes may vary depending on the region of a cave in which the animals are being mon- itored. Estimates of age and developmental rates of cave crayfishes suggest that they de- velop very slowly and that some can live for long periods (—30 years) (Cooper 1975; Coo- per and Cooper 1978; Weingartner 1977), but direct measurements over this long have not been made. In addition, since temperature is a regulating environmental factor in crusta- cean development, generalizations for all cave crayfishes are impractical since the animals in- habit various physiomorphic regions of caves. Here we report on a preliminary study to monitor the growth and laboratory mainte- nance of a stygobitic species (Orconectes aus- tralis packardi Rhoades, 1944) and an epigean (surface-dwelling) species (Orconectes crista- varius Taylor, 2000). We used both juvenile and mature crayfish so we could determine if various age groups would survive in controlled conditions. In this preliminary study, we re- port on the laboratory conditions used and the growth of these species maintained over a 1- year and in some cases a 2-year period in a defined laboratory setting. MATERIALS AND METHODS Orconectes a. packardi were collected in the Sloans Valley Cave System at die "Appa- lachian Trail" in a pool of water measuring 3 m wide, 6 m long, and at most 0.3 m deep. Orconectes cristavariiis were obtained from a relatively fast-moving surface stream (Four- mile Creek) by Fourmile Road in Clark Coun- ty, KY. Species identification was confirmed by Dr. Gunther Schuster of Eastern Kentucky University for O. cristavarius and by the tax- onomic key provided in Hobbs Jr. et al. (1977) for O. a. packardi. The crayfishes were transported to the lab- oratory in Lexington, KY, in water obtained from their environment. They were then transferred to individual aquaria (33 X 28 X 23 cm; water deplli 10—15 cm) and held as isolates throughout the study. Some individu- als were housed successfully for a year. Con- tainers were cleaned biweekly and animals were led with c< ercial lish food pellets (Aquadine), which is marketed as "shrimp and plankton sticks: sinking mini sticks." Since lliis consists of ground-up fish it would appear to be a suitable and nutritious diet for crayfishes. Fragments of cleaned chicken egg shell also were placed in the containers as a source of calcium. The chloramines Lexington uses for water purification were removed by carbon- based filters for the aquaria water. The car- bon-filtered water was held in a 190 liter (50 gallon) plastic tank and aerated for several days before utilization. Bacteria and algae were allowed to grow in the tank in order to detoxify any NH4+ and convert it to nitrite and nitrate, since NH.,+ is known to be toxic to crustaceans (McRae 1999). Cave animals were maintained in total darkness except for feed- ing, cleaning, and measuring. Epigean animals were exposed to a low light level widi a light cycle of 16:8 (light: dark) produced by full spectra lights (General Electric). When the aquaria were cleaned or when measurements were obtained, observations were made if the animals had molted by the appearance of the animal and by the presence, in the holding tank, of the chelae from the old exoskeleton. Neither species would fullv consume the che- lae after they consumed the rest of the old exoskeleton. The temperature was maintained at 16 to 17°C throughout the year; this was the temperature of the laboratory. At one pe- riod the laboratory temperature was uncon- trolled as mentioned in the results during the second year of this study. Aquaria for both the surface and cave species were stored in the same room over the same period ot time, and the tanks containing the cave species were covered with black plastic to Mock light. Aquaria were marked with numbers so that individuals could be monitored. Total body length (tip of rostrum to the end of telson) was measured to an accuracy ot 1 millimeter with a flexible plastic ruler. Mea- surements were obtained lour times during the first year. Alter the first year we learned thai thi' total body length measure is likely in- 110 Journal of tlie Kentucky Academy of Science 65(2) accurate because of the flexibility of the joint- ed abdomen. So for a subsequent year of hold- ing some of the same crayfishes in captivity, along with the addition of new individuals, the postorbital carapace length (from the posteri- or, dorsal surface of the orbital cup to the end ol the carapace directly posterior to die eye cup) was used. These measures were made with calipers (Swiss Precision, Switzerland, 0.1 mm). As for the first-vear study, four different time points were used throughout the year. A similar periodic sampling had previously been used by Weingartner (1977). The percent growth was determined by: [(postmolt length — premolt length)/(premolt lengdi)] X 100. RESULTS The epigean and stygobitic species grew and molted in captivity, but the survival rate over die first and second years was lower for the epigean species than for the stygobite. The highest mortality for the epigean species oc- curred after 7 mondis, although two of them died after 4 mondis. The stygobitic species demonstrated a better survival rate, with onlv one dying after 4 months and another after 9 months. The one that died at 4 mondis ap- peared to do so during ecdysis since the exu- vium was still attached to the body. The indi- viduals diat died are represented in Figure 1 as line plots that do not fully extend to the end of 1 or 2 years. The lines terminate at the period when measurements were last ob- tained. The frequency of molting was substantially higher (9 of 12 animals) for the epigean spe- cies within die first few months of contain- ment. A second molt was noted for only one of the epigean species within the first 12 months (Figure IB, note two asterisks). Of the four surface crayfish tiiat progressed to die second year of study onlv die largest one died. Three of the 13 stygobites molted during die same first few mondis and two of diem after 9 months. The individuals that molted are in- dicated widi asterisks witiiin the period of time a molt was noted (Figure 1). Only in die stygobitic species was it observed that some individuals actually had a reduction, instead of an increase, in their body length after a molt. This phenomenon has also been reported for O. a. australis by Cooper (1975), and in an earlier report Creaser (1934) stated that in the A Oap 6.0 E 5.5 -c 5.0 S 4.5 f 4.0 CD 3.5 - 3.0 100 — 1 — 200 300 — 1 400 B 7.5 7.0 E 6.5 - o ^ 6.0 - c 5.5- 0) t. 5.0- £ 4.5 H 4.0 - 3.5 O.c. 25^ 100 200 Days 300 400 Figure 1. Growth of stygobitic (Orconectes australis packardi) and epigean (Orconectes cristavarius) crayfishes in laboratory conditions. Growth curves were obtained by measures of total bodv length of the animals maintained as isolates (A) Orconectes australis packardi (O.a.p.) and (B) Orconectes cristavarius (O.c.) for a 1-year period in captivity. Asterisks denote that a molt occurred in that time period. Individuals diat died have their growth plot terminated at the last date a measurement was taken. There was substantial mortality for the epigean O.c. spe- cies after 7 mondis of containment. Initially N = 13 for O.a.p. and N = 12 for O.c. crayfish O. propinquus (Creaser used the name Faxonius propinquus) growth was not always associated with a molt when the male changed in sexual form. We are fairly confi- dent in the recorded dates of molts for each animal; however, there were a few animals, in both species, that indicated a growth in body length without a molt being observed. This lack in changes of lengtii we account to not being able to fully stretch the abdomen of the animal consistently during all measures. Per- haps the muscles between the segments of the abdomen were relaxed more during some Laboratory Growth ol Cave and Surface Crayfishes — Cooper mid ('oojicr 11 measures than others. This problem was avoided during the second year ol this study in which the postorbital carapace length was measured. Four freshly caught stygobites were added to the study for the second year because we released a tew from the first-year study back in the cave since we clipped one pair of an- tennules from some individuals at the end of the first-vear study to aid in another study in antennule growth within a molt cycle. Figure 2A contains the data obtained during the sec- ond year of the study of the stygobites. The ones in the graph depicted with circles were from the first-year studv and the ones repre- sented by triangles were crayfish newly added to the study. The results obtained in the sec- ond year indicate that the animals did increase in length during a molt although a minor amount in some cases. For the stygobites, three out of die four added to the study in the second year molted within the first 2 months. Six out of the eight molted within the last 150 days, which was likely a result of the labora- tory having environmental temperature swings for a period of about 2 weeks with tempera- tures reaching as high as 28°C. The few- epigean crayfish appeared to in- crease in size to a greater degree than die sty- gobitic species. As in Figure 2A, the lines in Figure 2B depicted with circles were ones from the first-year studv, and the crayfish rep- resented bv triangles were newly added epi- gean species. Only three animals were carried over from the first-year study. One died within 1 month, another within 3 months, and the last one within 7 months of the second studv in which postorbital carapace length was mea- sured. As in the first-year studv; the newly added epigean crayfish also showed a low re- sistance to laboratory rearing since a good number had died before the year was com- pleted. To determine die extent of die difference in growth between the two species during the first year of the studv. a percent change for each individual was determined and the means of the percent differences were com- pared between species (Figure 3A,). Since so many animals died in the epigean population in the period between 220 days and 365 davs, the 220-dav measurement session was used to calculate the percent difference in growth for 0) o 32 30 28 26 24 22 20 18 16 14 Oc. — i — 100 1 — 200 Days 300 400 Figure 2. Growth curves of stygobitic (Orconectes aus- tralis packardi) and epigean (Orconectes cristavarius) crayfishes as measured by postorbital carapace length. Plots for Orconectes australis packardi (O.a.p.) (A) and Orconectes cristavarius (O.c.) (B) are shown. Animals held throughout a second vear in captivity (solid circles) as compared to 1 year (solid triangles) are indicated. As- terisks denote that a molt occurred in that time period. The indhiduals that died have their growth plot termi- nated at the hist date a measurement was taken. As in the first vear of studv, diere was again high mortality for O.c. No O.c. survived for 2 years in captivity. Initially N = 13 for O.a.p. and N = 12 for O.c. this species during the first vear. Most of the growth for the epigean species occurred ear- lier than that for the stygobitic species. As mentioned earlier, the total body length was likely subject to error, but since such measures of surface crayfish species are rapidly made for aquaculture purposes, we retained these data since they can be of use. The growth measures during the second vear for the stygobitic and epigean crayfishes of the postorbital carapace 112 il of the Kentucky Academy oi Science 65(2) were also compared as a mean of a percent change (Figure 3B L One group of stvgobite crayfish was used for comparison of postorbital carapace length to total bodv length (Table 1). The postorbital carapace length accounts for about 40% of the total body length. The percent in the increase of growth, as measured bv postorbital cara- pace length, is ca. 9% (Table 2). There did not appear to be a correlation with size of the an- imal and the percent increase in growth within a single molt. DISCUSSION The results of this studv demonstrate that O. a. packardi can be maintained well in cap- thitv under defined conditions for up to 2 vears. This was the case for both small and large cra\fish. However, O. cristavarius ini- tially showed a good survival rate, which de- clined rapidlv die longer diey were housed, particularlv after 7 months. Since small and large epigean cra\-fish died, the failure to sur- vive was probablv due to unsuitable conditions for this species within the laboratory rather than to senescence. This studv also demon- strated that the epigean as well as the cave cra\fish molted and grew in captivity. One ap- proach to measure growth rates in crustaceans is to measure the time from one molt to the next in addition to size increases resulting from each molt. Only some of die cra\fish we held molted a second time (Table 3). Based on die molt frequency, diere is little or no dif- ference in growth rate between the two spe- cies, with O. a. packardi having slightly more second molts. In some cases where a molt was documented the cave animals did not show more dian a few millimeters increase in bodv length as assessed during the first year of studv bv total bodv lengtii. With measures of post- orbital carapace length, as used in die second year of studv. an approximate increase in lengtii widi each molt is 9%, but there is con- siderable individual variation (Table 2). Preliminary growth studies widi die stygo- bitic crayfish Orconectes inermis inermis Cope 1872. in Pless Cave, Indiana, using a mark- and-recapture approach, indicated diat juve- niles showed larger increments of growth at ecdvsis than mature animals (Hobbs III 1976). Using repetitive recaptures of hundreds of marked individuals of three species of stygo- £ 8 CO c 220 Days A2 8 - 730 Days I B Q. — t- <1> .E o 20 -, 18 16 14 12 - 10 8 6 - 4 - 2 0 O.a.p. 356 Days _L_ O.c. O.a.p O.a.p. O.c. Figure 3. Comparisons in the growth between stvgobitic (Orconectes australis packardi) and epigean (Orconectes cristavarius) cra\"fishes. (A,) During the first vear of studv in which die total body lengths were used, a comparison in percent of growth between Orconectes cristavarius (O.c.) and Orconectes australis packardi (O.a.p.) is shown (N = 13 for O.a.p. and N = 10 for O.c). Since so manv O.c. died, die values obtained after 220 days were used for both species. The error bars represent die ± of the standard error of the mean. (A,) The percent of growdi for O.a.p. based on total bodv lengtii, is shown after a 2- vear period in captivity. Since no O.c. survived for 2-vears a comparison could not be made. (B) In die second vear of studv the postorbital carapace lengtii was used as an index of growth (N = 8 for O.a.p. and N = 3 for O.c). The sample size is smaller for O.c. because of die lack in survival of this species. bitic cra\"fish in Shelta Cave, Alabama, Cooper (1975) predicted that one of die species, Or- conectes australis australis (Rhoades 1941), might five for 30 years or more. This was based on knowTi minimum size at recruitment, maximum size, and growdi increments ob- served following molts (Cooper and Cooper 1978, 1979: Culver 1982, p. 51). Detailed growth studies over a 1-year period of Cam- barus laevis from epigean groups as well as groups diat lived widiin die cave were con- ducted widi recapture techniques. From such measures, growth curves were established (Weingartner 1977). In Wemgartner's study a Laboratory Growth of Cave and Surface Crayfishes — Cooper and Coopi i 113 Talilr I. Comparison of total liody length to postorliital carapace length for Oreonietis anstralis packardi. Tuo ad- ditional animals not used in the growth studies were used for these morphometric measures. Table 2. The percent change in POL within .1 moll fc Oreonectes australis packardi. Premoll (n Total body Postorbitnl carapace Ratio length (1 length (POL) (mm) body/POL 43 16 0.37 44 IS 0,11 47 17.5 0.37 50 20 0.40 51 20 0.39 55 22.5 0.41 58 19 0.33 60 23 0.38 fid 23.5 0.39 71 29 0.41 71 28 0.39 Mean 0.39 Mean SEM 0.024 SEM 17.1 17.5 18.0 18.2 LS.3 19.0 20.0 20.5 21.0 22.8 23.5 26.4 27.0 28.0 ■ si Il 'II ISO L9.1 20.0 20.0 20,5 21.0 22.2 22.1 23.0 23,5 25.3 28.0 29.0 30.0 5.26 9 I 1 II. 1 9.S9 12.02 10.53 11.0 7.8 9.52 3.07 7.06 3.06 7.41 7.14 8.4 0.7 few animals were recaptured after 3 vears and the growth curves were extended for that length of time; however with only a few ani- mals the variability was not able to be assessed for differences in the age of the animals. Poor survivorship for O. cristavarius in our studies is likelv a species-dependent phenom- enon, since anodier epigean crayfish, Procam- barus clarkii (Girard 1852), from Raceland, Louisiana, survived well in die same labora- tory conditions for 2 vears or more. The rea- son for the difference in survival between these two epigean species mav be a result of emironmental adaptation, since O. cristavar- ius is predominantly found in fast-moving, highly oxygenated streams, and P. clarkii comes from swamps. It might be that die P. clarkii and O. a. packardi can survive well in water that is not highly oxygenated, such as that used in our laboratory, while O. crista- varius cannot. It is known that P. clarkii is ver\- J hardy and can tolerate all but the severest cas- es of hvpoxia (McClain 1999). There might also be dietary factors that we did not inves- tigate to account for die survival differences. All crayfishes held in captivity were fed die same diet at the same time. In addition, larger animals had a greater quantity of food provid- ed to them. Stygobitic crayfishes also show an amazing resistance to experimentally induced fluctua- tions in temperature, from freezing in blocks of ice to rapid exposure of high temperature (32.5°C) (Park et al. L941). In addition, they are known to be starvation resistant, possibly due to a lower metabolic rate (Dickson and Franz 1980; Dickson and Giesy 1982). Bur- banck et al. (1948) and Jegla (1964) both re- ported diat cave crayfish have a lower meta- bolic rate as compared to epigean species. Comparable studies on O. cristavarius are lacking, so it remains unknown if this species can tolerate general stress as well as P. clarkii and O. a. packardi. The differential in the initial growth rate between O. cristavarius and O. a. packardi is interesting. It is possible that the handling, transport, and exposure to a new environment are factors, although care was taken not to al- low the animals to heat up in transport or be exposed to fluctuating temperature. However, a change in temperature from a surface stream or cave could have an impact on the initial molting frequency. Temperature is con- Table 3. Comparison of growth rates by frequency of molts within the first and second years of captivity for Oreonectes australis packardi and Oreonectes cristavarius crayfishes. 1st molt 2nd molt Isl year O.a.p. O.e. 5 of 12 animals LOol 12 animals none 1 of 4 2nd year 0.a.p. O.e 14 of S animals 10 of 9 animals 5 of 8 animals 3 11I 3 animals 114 ial of the Kentucky Academy <>l Science 65(2) sidered to be one of the most significant lac- tors in regulating crustacean growth (Conan 1985): not only an increase in water temper- ature but a reduction as well can induce molt- ing. Other small crustaceans, such as copepods (Vidal 1980) and amphipods (Dagg 1976), re- duce their molt frequency in cold tempera- tures, but when this occurs the animals grow larger after each molt than when they molt more frequently. In contrast, an increase in temperature can inhibit some crustaceans from molting (Haefner and van Engle 1975). The shrimp Crangon crangon Linnaeus is known to have varied longevity depending on environmental temperature (Labat 1977; Llyod and Yonge 1947; Oh 1999). For a review of factors regulating growth in crustaceans see Wenner (1985). The increase in initial growth of O. crista- varius is not likely due to 'catch-up' growth, common in crustaceans when they are ex- posed to sufficient food after being deprived of food (Bostworth and Wolters 1995), since the habitat from which they were obtained was abundant in crayfish, fish, and snails as dietary resources. On the other hand, the lack of growth for O. a. packardi even after a molt might be expected since it has been reported that an O. a. australis measuring 35.2 mm in total carapace length did not alter its length after two molts (Cooper 1975). It would be of value to know if stygobitic crayfish do show 'catch-up' growth depending on resources, or if the growth attained for a molt is tempera- ture regulated. If such growth does occur it would make it difficult to determine the cor- rect age of adult animals without knowing their complete life history and environmental conditions. This is particularly relevant since Weingartner (1977) showed differential rates of growth within various regions in a single cave system. It was concluded that the envi- ronmental differences in water temperature and food resources likely accounted for the re- gional differences. It is our hope that one will consider the pos- sibility of long-term rearing of crayfishes in suitable laboratory conditions to gain insight into environmental factors regulating the de- velopmental issues and to further refine lab- oratory conditions that promote survival. ACKNOWLEDGMENTS We thank Richard M. Cooper for his help with feeding and maintaining the Crayfish; John LaMar Cole and Dr. Milan L. Hopper (University of Kentucky) for help in obtaining cave crayfish and bearing with us on the time- consuming cave expeditions in the Sloan's Val- ley Cave System where die cave crayfish were obtained; and Hyewon Cooper and the Kim family of Lexington for helping to obtain epi- gean crayfish. Editorial assistance was provid- ed by Austin M. Cooper. LITERATURE CITED Barr, T. C, Jr., and R. A. Kuehne. 1971. Ecological studies in the Mammoth Cave system of Kentucky II. The eco- system. Annales Speleol. 26(l):47-96. Bostworth, B. G., and W. R. Wolters. 1995. Compensatory growth in juvenile red swamp crawfish, Procambarus clarkit. Pages 64S-656 in R. P. Romaire (ed). EighrJi International Symposium on Astacology. Louisiana State University, Baton Rouge, LA. Burbanek, W. D., J. P. Edwards, and M. P. Burbanck. 1948. Toleration of lowered oxygen tension by cave and stream crayfish. Ecology 29:360-367. Conan, G. Y. 1985. Periodicity and phasing of molting. Pages 73-99 in A. M. Wenner (ed). Crustacean issues 3, Factors in adult growth. AA. Balkema, Boston, MA. Cooper, J. E. 1975. Ecological and behavioral studies in Shelta Cave, Alabama, with emphasis on decapod crus- taceans. Ph.D. Thesis. University of Kentucky, Lexing- ton, KY. Cooper, J. E., and M. R. Cooper. 1976. Marking crayfish for long-term ecological studies. ASB Bull. 23(2):52. [abstract]. Cooper, J. E., and M. R. Cooper. 1978. Growth, longevity, and reproduction strategies in Shelta Cave crayfishes. NSS Bull. 40(3):97. [abstract]. Cooper, J. E., and M. R. Cooper. 1979. Growdi, longevity, and reproduction strategies in Shelta Cave crayfishes. Page 85 in E. Werner (ed). Proceedings of the 1977 National Speleological Society Annual Convention, Al- pena, MI. Cope, E. D. 1872. On die Wyandotte Cave and its fauna. Am. Naturalist 6:406-422. Creaser, E. P. 1934. Age, growdi, and sex ratios in die crayfish Faxonius propinquus. Trans. Michigan Acad. Sci. 14:581-585. Culver, D. C. 1982. Cave life: evolution and ecology. Har- vard Univ. Press, Cambridge, MA. Dagg, M. J. 1976. Complete carbon and nitrogen budgets for the carnivorous amphipod, Calliopus laeviusculus (Kroyer). Int. Rev. Ges. Hydrobiol. 61:297-357. Dickson, G. W, and R. Franz. 1980. Respiration rates, ATP turnover and adenylate energy charge in excised Laboratory Growth ol Cave and Surface Crayfishes — Cooper and ('nop I gills of surface and cave crayfish. ( !omp. Biochem, Phy- siol. 65 A:375-379. Dickson, C. W., and J. P. Giesy. I9S2. The effects of star- vation on muscle phosphoadenylate concentrations and adenylate energy charge of surface and cave crayfish. Comp. Biochem. Physiol. 71 A:357-361. Fingerman. S. W. 1985. Non-metal environmental pollut- ants and growth. Pages 219-234 in A. M. W'enner (ed). Crustacean issues 3, Factors in adult growth. AA. Bal- kema, Boston, MA. Girard, C. 1852. A revision of the North American Astaci, with observations on their habits and geographical dis- tribution. Proc. Acad. Nat. Sci. Philadelphia 6:87-91 Ilaefner, P. A., Jr., and W. A. van Engel. 1975. Aspects of molting, growth and survival of male rock crabs Cancer irroratus, in Chesapeake Bay. Chesapeake Sci. 16:253- 265. Hobbs, 11. 11.. Jr., II. 11. llobbs. III, and M. A. Daniel. 1977. A review of the troglobitic decapod crustaceans of die Americas. Smithsonian Contrib. Zool. 244:1-183. Hobbs, II. II., III. 1973. The population dynamics of cave crayfishes and their commensal ostracods from south- ern Indiana. Ph.D. Thesis. Indiana University, Bloo- mington, IN. Hobbs, H. II., III. 1976. Molt cycle, size and growth in Orconectes inermis inermis Cope (Decapoda: Cambar- idae). Virginia J. Sci. 27:44. [abstract]. Hobbs. II. II., III. 1978. Studies of the cave crayfish, Or- conectes inermis inermis Cope (Decapoda, Cambari- dae). Part IV: Mark-recapture procedures for estimating population size and movements of individuals. Int. J. Speleol. 10:303-322. Jegla, T. C. 1964. Studies of die eyestalk, metabolism, and molting and reproductive cycles in a cave crayfish. Ph.D. Thesis. University of Illinois, Champaign, IL. Jegla, T. C. 1966. Reproductive and molting cycles in cave crayfish. Biol. Bull. 130:345-358. Jegla. T. C. 1969. Cave crayfish: Annual periods of molting and reproduction. Actes du IV' Congr. Int. Speleol. Yu- goslavie (12-26 IX 1965) 4-5:135-137. Labat, J. P. 1977. Ecologie de Crangon crangon (L.) (De- capoda, Caridea) dans un etang de la cote langue-do- cienne. Vie et Millieu 30, ser. A:232-292. Llvod, A. J., and C. M. Yonge. 1947. The biology of Cran- gon vulgaris L. in the Bristol channel and Severn es- tuarv. J. Mar. Biol. Assoc. United Kingdom 26:626-661. McClain, VV. R. 1999. Effects of hypoxia on growth and survival ol the crayfish Procambams clarkti. Freshwater Crayfish 12:121 L33 MeRae. T. L999. Chemical removal of nitrite and chlori- nating agents froi unicipal water supplies used lor crayfish and aquarium finfish culture. Freshwatei ( Iraj fish 12: 727-732. oli. C.-W. 1999. Life history and population dynamics ol the crangonid shrimps Crangon crangon and PhUoch- eras trispinosus in Port Erin Bay. Isle of Man. Ph.D. Thesis. University of Liverpool, Liverpool, UK. Packard. A. S. 1888. The cave fauna of North America with remarks on the anatomy of the brain and origin of the blind species. Mem. Natl. Acad. Sci. 4(1):] 156, 26 plates. Park, O.. T. W. Roberts, and S. J. Harris. 1941. Prelimi- nary analysis of activity of the cave crayfish, Cambarus pellucidus. Am. Naturalist 75:154-171. Poulson, T L. 1964. Animals in aquatic environments: an- imals in caves. Pages 749-771 in D. B. Dill, E. F. Adolph, and C. G. Wilber (eds). Handbook of physiol- ogy. Section 4, Adaptations to the environment. Wil- liams and Wilkins Co., Baltimore, MD. Poulson, T. L. 1992. The Mammoth Cave ecosystem. Pag- es 569-611 in A. I. Camacho (ed). The natural history of biospeleology. Monogr. Natl. Mus. Nat. Sci. Madrid. Rhoades, R. 1941. Notes on some crayfishes from Ala- bama caves, widi the description of a new species and a new sub-species. Proc. U.S. Naturalist Mus. 91:141- 148. Rhoades, R. 1944. The crayfishes of Kentucky. Am. Midi. Naturalist 31:111-149. Siegel, P. R., and A. M. Wenner. 19S5. Field and labora- tory studies of sand crab growth. Pages 165-170 in A. M. Wenner (ed). Crustacean issues 3, Factors in adult growdi. AA. Balkema, Boston, MA. Taylor, C. A. 2000. Systematic studies of the Orconectes juvenilis complex (Decapoda: Cambaridae), with de- scriptions of two new species. J. Crust. Biol. 20:132- 152. Vidal, J. 1980. Phvsioecology of zooplankton I. Effects of phytoplankton concentration, temperature, and body size on the growth of Calanus pacificus and Pseudoca- lanus sp. Marine Biol. 50:111-134. Weingartner, D. I. 1977. Production and tropic ecolog) "I two crayfish species cohabiting an Indiana cave. Ph.D. Thesis. Michigan State University, East Lansing. Ml Wenner, A. M. 1985. (ed). Crustacean growth: Factors in adult growth. Crustacean issues 3. AA. Balkema, Bos- ton, MA. J. Ky. Acad Sd 65 2 :116-131. 2004. Vascular Flora of Five Reservoirs in the Berea College Forest, Madison and Jackson Counties, Kentucky Ralph L. Thompson Herbarium. Biologv Department. Berea College, Berea, Kentucky 40404-2121 and Chris A. Fleming Department of Botany, University of Tennessee, Knoxville, Tennessee 37996-1100 ABSTRACT A descriptive survey of the vascular plants from wetland and aquatic habitats of the five Berea College reservoirs, Madison and Jackson counties, in south-central Kentucky was conducted during the growing seasons of 1995, 1996, 1998, and 2003. Six plant habitats described are Vegetated Open Water, Emergent Marsh, Shrub Swamp. Seasonally Dewatered Mud and Sandflat, Sedge-grass Meadow, and Shallow' Stream with Gravel Bar. Relative abundance values were determined for every taxon at each reservoir. The species list comprises 292 species, 175 genera, and 65 families. Thirty-eight taxa (13%) are exotics. Species are classified as Lycopodiophyta (1). Equisetophyta (2), Polypodiophyta (7), Pinophyta (1), and Magnolioph\ta (2S1). In the National Wetland Classification, 205 species (70.0%) are obligate, facultative wedand. and facultative species. INTRODUCTION The Berea College Forest (BCF) consists of 3318 ha (8200 acres) in several tracts in parts of Madison, Jackson, and Rockcastle counties of south-central Kentucky. BCF is maintained for watershed/wilderness resources, timber management, wildlife management, recrea- tion, and educational purposes by Berea Col- lege, Berea, Kentucky. In addition, the BCF has four reservoirs in Madison and Jackson counties diat pro\ide the water supplv for die citv of Berea and surrounding Madison Coun- tv communities and one reservoir for flood- prevention purposes. Prior to 1904, people from Berea obtained their water from wells or cisterns. In 1904, Berea College began collecting water from springs in die Berea College Forest and start- ed die Berea College Waterworks. In 1919, Upper Silver Creek Reservoir was built to col- lect and store water for the citv. A second res- ervoir. Lower Silver Creek Lake, was created in 1939 to pro\ide more water. Cowbell Lake was built in 1954 because of increased water demand bv Berea and soudi Madison Counts". As more water continued to be needed for Be- rea, its industrial areas, and south Madison County, Owslev Fork Reservoir was construct- ed in 1976. In 1986, Berea College was re- quired to build an earthen dam to serve as a flood-retarding structure for the overflow or breach of the Cowbell Reservoir dam. This ac- cumulated bodv of water is known as Red Lick Reservoir No. 2. These five reservoir lakes are managed and regulated bv Berea College Util- ities. Ferren and Tonsor (1996) defined a reser- voir as a pond or lake, natural or artificial, from which water may be withdrawn for irri- gation and/or water supplv. Our floristic studv is a baseline inventory with emphasis on die wetland and aquatic flora from collections and observations at five man-made bodies of water, or lakes, which we have termed "reservoirs" for descriptive purposes. These bodies of wa- ter are classified under the Lacustrine System of Cowardin et al. (1979), which are charac- teristically bounded bv uplands or bv wetlands dominated by shrubs, persistent emergents, and emergent mosses and lichens. Published botanical works for the Berea College Forest are Grossman and Pittillo (1962), Wade and Thompson (1990), and Thompson and Fleming (2004). Among diese three studies, over 600 plant species have been collected from die forest. Recent wet- land floristic studies in soudi-central and east- central Kentucky include Hoagland and Jones 116 Flora <>l Five Reservoirs in the Berea College Foresl — Thompson and Fleming, I I Kentucky Figure 1. The five reservoirs within the Berea College Forest. Bighill Quadrangle. Kentucky, Photon-vised 1979: 1 Upper Silver Creek Reservoir, (2) Lower Silver Creek Reservoir, (3) Cowbell Reservoir, (4) Owsley Fork Reservoir, and (5) Red Lick Reservoir No. 2. Kentucky inset with Madison (upper left) and Jackson (lower right) counties dark- ened. (1992), Luken and Bezold (2000), and Thompson and FitzGerald (2003). Baseline inventories of the vascular flora from reservoirs are important for preserving wetland areas, moderating the effects of floods, improving water qualitv. and enhancing aesthetic and heritage value (Mitsch and Gos- selink 1993). Wetland studies have provided valuable information on soils, hvdrological pat- terns, plant communities, hvdrosere succes- sion, and environmental controlling factors (Meagher and Tonsor 1992). The objectives of our descriptive floristic survey were (1) to document the vascular flora vvitli representative voucher specimens, (2) to ascertain plant origin (i.e., native or exotic), (3) to indicate National Wetland Classification Status for each taxon, (4) to determine relative abundance, and (5) to describe habitats for each species. THE ENVIRONMENTAL COMPLEX Reservoir Descriptions Upper Silver Creek Reservoir (USC). Also known as Kale Lake or A-Lake, USC is the smallest reservoir, 2.1-ha surface area (5.3 acres), and was formed by damming die East Fork of Silver Creek. The concrete dam lies at 341 m elevation. USC is 10 m deep at its deepest point with visibility of 3.5 m. Upper Silver Creek reservoir is located in Madison County at the end of a gravel road 2 km soudi of KY 21 and 5 km west of the US 421 junc- tion with KY 21 at Bighill (Figure 1). Lower Silver Creek Reservoir (LSC). LSC (B-Lake or North Lake) has 7.8-ha surface area (19.3 acres). The north-trending concrete dam spillway lies at 29S m elevation. LSC is 10.5 m deep with visibility at 3 m. Lower Sil- ver Creek Reservoir lies 0.3 km north of USC on the East Fork Silver Creek and 1.7 km south of KY 21 (Figure 1). Coicbell Reservoir ( CRR ). CBR has 6.8-ha (16.8 acres) surface area and was formed by damming of the Cowbell Creek watershed. A north-trending earthen and concrete dam lies at 301 m elevation. CBR is 12 m deep with visibility of 3 m. Cowbell Reservoir is located in Madison County at the end of a gravel road 1.8 km south of KY 21 and 2.4 km west ol junction US 421 and KY 21 at Bighill (Figure 1). 118 Journal of the Kentucky Academy oi Science 65(2) Owsley Fork Reservoir (OFR). This, tin- largest reservoir, has 61.1-ha (151 acres) sur- face area. OFR was formed by damming Ows- ley Fork Creek and Radford Hollow tributary. The north- and northwest-trending earthen and concrete dam lies on the Madison and Jackson count)' boundary at 250 m elevation. OFR has a maximum depth of 12 m with vis- ibility of 3.5 m. Owsley Fork Reservoir is the only lake open to the general public, but with certain restrictions. OFR is located 4 km east of Bighill from the junction of US 421 with KY 21 which becomes Owsley Fork Road on the east side of OWF in Jackson County. A paved road, Radford Hollow Road, separates from Owsley Fork Road to the southwest of the dam on the Madison County side (Figure 1). Red Lick Reservoir No. 2 (RLR). This res- ervoir of 3-ha (7.4 acres) was designed as a flood-retarding structure with an earthen dam formed from landfill from the adjacent mixed hardwood forest. The elevation of the dam top is 262 m. RLR is 5 m deep with limited visi- bility of 1 m. It is located 1.2 km north of Cowbell Reservoir on Cowbell Creek, and 0.7 km south of KY 21 (Figure 1). Physiography and Geology The Berea College Forest of Madison and Jackson counties lies within the Knobs Lower Scioto Dissected Plateau Region of the West- ern Allegheny Plateau and the Knobs Normal Upland Region of the Interior Plateau (Wood et al. 2002). The geological substrate within the area of the Berea College reservoirs is very complex. Drainage below the reservoirs has some Holocene Alluvium (210-230 m) of the Quaternary System, and the valleys have some New Albany Shale (230-275 m) of the Upper Devonian and Lower Mississippian System. Bedrock surrounding the reservoirs on lower slopes consists of shale, siltstone, and lime- stone of die Nancy Member (275-335 m), Cowbell Member (335-365 m), Nada Mem- ber (365-385 m), and Renfro Member (385- 396 m). All these members belong to the Bor- den Formation of the Mississippian System (Weir et al. 1971). The middle and upper slopes and ridgetops of the Knobs consist of limestone and shale of Newman Limestone Member (396^50 m) and Pennington For- mation (450-457 m) sandstone, siltstone and shak', both of the Upper Mississippian Sys- tem. Several higher Knobs are capped with the Livingston Conglomerate Member (457- 470 m) and/or the Corbin Sandstone Member (470-488 m) in the Lee Formation of the Lower Pennsylvanian System (Weir et al. 1971). Soils The majority of the forest soils surrounding the reservoirs belong to the the Weikert series or the Bledsoe-Gilpin-Shelocta-Grigsby series. The Weikert series are shallow, well-drained, moderately permeable soils with an acid pH from 4.5 to 5 derived mainly from acid silt- stones (Newton et al. 1973). The Weikert soil series includes the majority of the soils sur- rounding Cowbell, Red Lick Flood Retarding Structure, Upper Silver Creek, and Lower Sil- ver Creek reservoirs located entirely in Mad- ison County. The soils of the Bledsoe-Gilpin- Shelocta-Grigsby series are typically deep, well-drained, and moderately permeable soils with a pH from 4.6 to 7. They are formed mainly in mixed alluvium from acid siltstones and shales (Hayes 1989). This soil association is found only at the Owsley Fork Reservoir study site in Jackson and Madison counties. Climate The climate of Kentucky is warm temper- ate, humid mesothermal, with little or no wa- ter deficiency, and is characterized by long warm summers and short mild winters (Tre- wartha and Horn 1980). The mean annual temperature of Berea is 13.7°C. July and Au- gust are usually the warmest months, with a mean temperature of 24.6°C, and January is the coldest mondr, widi a mean temperature of 1.5°C. The mean annual precipitation is 119.3 cm widi July having the highest precip- itation at 12.2 cm and October the lowest with 5.8 cm. The mean frost-free growing season is 189 days with the mean date of die last spring freeze on 15 April and the first fall freeze on 22 October (Conner 1980). Vegetation Western Mesophytic Forest, a mosaic re- gion of Mixed Mesophytic Forest and Oak- Hickory Forest, is the predominant vegetation in The Knobs Border Area as described by Braun (1950). Muller and McComb (1986) Flora of Five Reservoirs in the Berea College Forest — Tl, wmpson nut I Fie classified the upland lorests Irom eight sites within the Kentucky Knobs Region into me- sophvtic hardwoods, white oak, chestnut oak, and scarlet oak forest types. The upland forest bordering the Berea College reservoirs on the north- and northwest-trending aspects is pri- marily a mosaic of mesophytic hardwoods (Liriodendron-Acer-Fagus-Quercus-Carya- Fraxinus). The forest stands on the east-trend- ing aspect and valley bottoms in the vicinity of the reservoirs are intermixed with mixed oak hardwoods-pine (Quercus-Pinus). All five res- ervoirs are surrounded by forests and open lands in various serai stages of secondary suc- cession. METHODS The vascular flora was collected from die wetland habitats of the five reservoirs through- out the growing seasons of 1995, 1996, 1998, and 2003. Plants not in wetland habitats (i.e., contiguous secondary successional ecotones, upland forests, and reservoir dams) were not included in the species list. Manuals used for plant identification were Gleason and Cron- quist (1991), Strausbaugh and Core (1978), and Beal and Thieret (1986). Plant classifica- tion and nomenclature are mostly from Glea- son and Cronquist (1991). Representative voucher specimens were processed according to standard herbarium procedures and depos- ited in the Berea College Herbarium (BE- REA). Previous plant collections from die BCF were also examined at BEREA. Six wetland habitats modified from Thomp- son and FitzGerald (2003) were delineated from field reconnaissance of wetland habitats and species composition in these wetland ar- eas from each reservoir. These wetland habi- tats are enclosed in brackets after the National Wetland Category in the Appendix. The six wetland habitats are (1) Vegetated Open Wa- ter (VOW) — an area of permanent water with a depth of up to 2 m with vegetation restricted to obligate free-floating species, floating- leaved submergents, and submergents; (2) Shrub Swamp (SS) — seasonally flooded area with saturated soils present when no standing water exists and dominated by riparian trees and shrubs with wetland herbaceous species; (3) Emergent Marsh (EM) — an area that is characterized by permanent or seasonal Hood- ing or by water letdown, resulting in saturated soils during dr\ summers, and thai supports obligate and facultative amphibious herba- ceous plants: (4) Sedge-grass Meadow (SGM) — an area that has typically saturated soils covered with shallow water during limes of increased rainfall and that supports obligate and facultative wetland herbs; (5) SeasonalK Dewatered Mud-Sand Flat (SDF) — an area that is inundated for most of the year but with water receding early enough in summer, sup- porting annual species along exposed mudflats or sandy shoreline. The boundary between the seasonally dewatered flats and emergent marshes may vary from year to year based upon precipitation, evaporation, and water drawdown; and (6) Shallow Stream with Grav- el Bar (SSG) — headwater inlet streams with vegetated streambanks, terraces, and gravel bar areas from periodical flooding; this habitat supports a rich assemblage of wetland and for- est perennial herbs and woody plants. The for- ests and open lands in various serai stages of secondary succession surrounding the reser- voirs were not inventoried for plants or in- cluded in the species list. The Appendix includes the species name, plant origin (native or exotic). National Wet- land Classification status, a representative wet- land habitat for each taxon, and the relative abundance value of each species found at each reservoir. An asterisk (*) preceding a species name denotes an exotic or non-indigenous plant spe- cies. After the species name is die National Wetland Category (Reed 1988). These Nation- al Wedand Categories are OBL = Obligate Wetland, FACW = Facultative Wetland, FAC = Facultative, FACU = Facultative Upland, and UPL = Upland, and may contain minus ( — ) or plus ( + ) designations for the drier or wetter limits of the facultative categories. A Not Categorized (NC) has been created for those species not listed or classified in die 1997 National Wetland Indicator Plant List, Northeastern Region 1 by the USFWS (1997) (Appendix). In the Appendix, abbreviations horizontally from the earliest to most recently established are reservoirs are: USC = Upper Silver Creek, LSC = Lower Silver Creek, CBR = Cowbell. OFR = Owsley Fork, and RLR = Red Lick Reservoir No. 2. Relative abundance values are listed under the columns lor each reser- 120 [oumal ot the Kentucky Academy <>i Science 65(2) Table 1. Classification of vascular plants at Five li voirs in tin- Berea College Forest. Kentucky. S|K-(_il-s composition Division Familii s Genera Species Native Exotic Equisetoph\ta 1 i 2 2 0 0.69 Lvcopodioplnta 1 i 1 1 0 0.34 Polypodiophyta 3 5 7 7 0 2.40 Pinophyta 1 1 1 1 0 0.34 Magnoliophyta 59 167 281 243 38 96.23 Masjnoliopsida 44 125 190 167 23 65.07 Liliopsida 15 42 91 76 15 31.16 Totals 65 175 292 254 38 100.00 voir. Relative abundance values adapted from Thompson and Jones (2001) are R = Rare — 1 to 5 indhiduals or colonies; I = Infre- quent— 6 to 30 indhiduals or colonies; O = Occasional — 31 to 100 indhiduals or colonies; F = Frequent — hundreds of indhiduals or colonies; and A = Abundant — thousands of indhiduals or colonies (Appendix). RESULTS Taxonomic Summarv The documented vascular flora of die five Berea College Reservoirs comprises 292 spe- cies widiin 175 genera from 65 families. The annotated catalogue of plant species is com- posed of Lvcopodiophyta (1), Equisetoplrvta (2), Pohpodiophxta (7). Pinophvta (1), and Magnoliophvta (281). Thirty-eight species (13%) are exotics (Table 1). Two hundred five (70.0%) are OBL, FACW, and FAC wedand species (Appendix). The number of species in die wedand categories are OBL (61), FACW (73), FAC (70), FACU (61). UPL (11), and NC (16). Total species for each reservoir are USC (114), LSC (177), CBR (150), OFR (214), and RLR (135) (Appendix). The largest families in species are Asteraceae (48), Po- aceae (35), Cvperaceae (24), Lamiaceae (13), Fabaceae (12), Polvgonaceae (12), and Jun- caceae (9). The largest genera are Polygonum (11), Juncus (9), Panicum (9), Aster (8), and Carex (8) (Appendix). Polygonum densiflorum is documented as a new Kentucky record. Reservoir Flora and Habitats The five reservoirs have developed various wetland habitats dirough hvdrarch or hvdro- sere succession. Characteristic species and lo- eallv rare taxa are listed for each reservoir hab- itat. There is some zone intergradation in the species from the six habitats, some more con- spicuous then others; e.g., a gradient or con- tinuum exists between species among the EM. SS, and SDF habitats at Owslev Fork Reser- voir. The flora and habitats of the five reservoirs in the Berea College Forest — Upper Silver Creek, Lower Silver Creek, Cowbell, Owslev Fork, and Red Lick Reservoir Xo. 2 — are de- scribed from die oldest to die earliest created bodv of water. Upper Silver Creek Reservoir. The four habitats of USC are VOW EM. SDF, and SGM. Important submerged vegetation in the VOW are Xaja.s guadalupensis. X. minor and Potamogeton nodosus. The EM lies in steep areas surrounding die reservoir and at a small area by die dam spillwav. Generallv, the EM and SDF habitats are hea\ilv shaded bv the encroaching upland vegetation. Characteristic species of die EM include Alisma subcorda- tum, Cyperus strigosus, Juncus acuminatu.s, J. effusu.s var. solutus, Leersia oryzoide.s, Lud- wigia alternifolia, Scirpus cyperinus, and Scu- tellaria lateriflora. The SDF occurs as small slough delta areas at die northwest shore and east shore, which are created from side ra\ine stream siltation and water drawdown. T\pical taxa of this hab- itat include Bidens spp., Boehmeria cylindrica, Impatiens capensis, Ludwigia alternifolia, Ly- copus virginicus, Mimulus alatus, and Pentho- rum sedoides. At die north end on die lower part of die dam, a small wet-meadow seepage exists. Some SGM species include Apocynum cannabinum, Carex frankii, C. lurida, C. vul- pinoidea, Eupatorium perfoliatum, E. serotin- um, Linum striatum, Polygala sanguinea, Scir- pus atrovirem, and S. pendulus. Scirpus po- lyphyllus is restricted to USC. The USC flora is represented bv 27 OBL. 27 FACW, 26 FAC. 25 FACU, 4 UPL, and 5 NC species. Lower Silver Creek Reservoir. The five wedand habitats of LSC are VOW, EM, SDF SS, and SGM. The VOW habitat contains Xa- jas guadalupensis, X. minor, and Potamogeton nodosus. The EM zone occurs at die north- western end by die concrete spillway, exten- sively at die southern end, and in small areas along the wTest-and-east trending banks. Char- Flora "I Five Reservoirs in the Berea College Forest— Thon vpson mid Fleming 121 acteristic emergents arc Eleocharis quadran- gulata, Juncus acuminatus, Juncus effusus var. solutus, Leersia oryzoides, Scirpus cyperinus, S. validus, and Typha latifolia. Other EM taxa include Acorus calamus, Asclepias incarnata, Carex spp., Hypericum mutilum, Ludwigiaal- ternifolia, L. palustris, Lycopus oirginicus, Onoelea sensibilis, and Panicum rieidulum. The SDF habitat nearly encompasses the entire reservoir during times of water draw- down except at the concrete dam. Character- istic species are Bidens spp., Boehmeria ci/lin- drica, Fimbristylis autumnalis, Impatiens ca- pensis, Lycopus oirginicus, Microstegium oi- mineum, Mimulus alatus, M. ringens, Polygonum spp., Scirpus atrovirens, and Xan- ihium strumarium. A small zone of SS behind the EM at the south end includes scattered Platanus occidentalis, Salix cxigua, S. nigra, S. sericea, and Sambucus canadensis. The her- baceous laver of SS mainly comprises species present in the seasonally dewatered flats and emergent marsh. The SGM habitats in die southern terminus and die northern end com- prise the most floristicallv rich habitat. Char- acteristic SGM species include Agrimonia parviflora, Asclepias incarnata. Aster dumo- sus, Carex frankii, C. lurida, C. tribuloides, C. oulpinoidea, Eupatorium coelestinum, E. jis- tulosum, E. perfoliatum, Euthamia gramini- folia, Helianthus angustifolius, Lobelia cardi- nalis, L. siphilitica, Panicum clandestinum, Poli/gala sanguinea, Rhcxia oirginica, and Spi- ranthes cernua. Wetland species found only at LSC are Aco- rns calamus. Aster puniceus. Carex squarrosa, C. stipata, Cyperus brevifolioides, Habenaria flava, H. lacera, H. pcramocna, Lilium cana- dense, Pluchea camphorata, and Solidago ru- gosa (Appendix). The LSC flora is represented by 36 OBL, 50 FACW, 40 FAC, 36 FACU, 5 UPL, and 10 NC species. Cowbell Reservoir. Four wetland habitats at CBR are classified as VOW, EM. SDF. and SS. The perimeter is entirely forested with steep terrain on both the east and west-trend- ing banks. Najas guadalupensis, Potamogeton diversifolius. P. nodosus, and P. pusillus, are found in the VOW zone. The EM at the south and southwest watershed area supports typical hvdrosere successional plants. Characteristic emergent species are Eleocharis quadrangu- lata, JunCUS effusus var. SolutUS, Leersia ory- zoides, Panicum rigidulum Scirpus cypt rinus S. pendulus, and Typha latifolia. The SDF zone is located around shoreline especiall) on the west and south sides. Characteristic spe- cies along the SDK shoreline are Boehmeria cylindrica, Carex vulpinoidea, Impatiens ca- pensis, I. pallida. Junius tenuis, Lycopus oir- ginicus, Polygonum cespitosum var. hngise- lum. Polygonum punctatum, and Rotala ra- mosior. Shrub swamp habitats are located on the east and west banks at CBR. Indicator wood) trees are Acer negundo, A. saccharinum, Bet- ula )iigra. Platanus occidentalis, Populus del- toides, and Salix nigra. Characteristic shrubs are Hydrangea arborescens, Lindera 1/cuzoin. and Sambucus canadensis. Toxicodendron racl- icans is the most prevalent woody vine. Her- baceous plants in the SS differ little in species composition from those found in the sur- rounding EM and SDF habitats. Wetland species, Betula nigra. Gratiola neg- lecta, Populus deltoides, Potamogeton diicrsi- folius. P. pusillus, and Thelypteris hexagonop- tera, are recorded only from CBR (Appendix). The CBR flora consists of 34 OBL. 35 FACW, 34 FAC, 35 FACU. 4 UPL. and 8 NC species. Owsley Fork Reservoir. All six wetland habitats— VOW, EM, SDF. SS, SGM, and SSG — are present at OFR. Characteristic spe- cies in the VOW are Najas guadalupensis, N. minor, Potamogeton illinoensis, and P. nodo- sus. Two large areas of EM are present at OWF; one located at die western shore of Madison County and the other at the south- eastern shore of Jackson County. Characteris- tic emergents are Alisma subcordatum. Eleo- charis palustris, E. quadrangulata, Equisetum arvense, E. hyemale, Galium tinctorium, jun- cus effusus var. solutus. Ludwigia alternifolia, Sagittaria australis. Scirpus purshianus, S. validus. Typha angustifolia, and T. latifolia (Appendix). The Seasonally Dewatered Mud-Sand Flats encircle the entire lake in the southwest and southeast coves of the reservoir where the shoreline is much more level, exposed, and drier during water letdown than the shaded north-trending aspect. Characteristic species of the SDF are Bidens spp.. Cyperus strigo- sus. Diodia oirginiana, Eclipta prostrata, Eleo- 122 Journal of the Kentucky Academy "I Science (i5 2 charts ovata, Lindernia dubia, Ludwigia pal- ustris, Lysimachia nummularia, Penthorum sedoides, Polygonum spp., Rotala ramosior, Scirpus purshianus, Scutellaria lateriflora, and Xanthium strumarium. Shrub swamp habitats serve as an intergrad- ing boundary between the SDF and EM zones in the largest coves of Owsley Fork Reservoir. Woodv species of the Jackson counts' SS in the eastern comer of the lake are Acer negundo, A. saccharinum, Cornus drummondii, Platan- us occidentalis, Salix exigua, S. nigra, and S. sericea. Herbaceous wetland species princi- pally include those of die SGM and SDF zones. The SS in the northwest corner of the reservoir in Jackson Counts' has nearly the same floristic composition in die Madison County side. Owsley Fork Reservoir has a large SGM on the north side in Jackson Counts' and also on die east side in Madison Count)'. The SGM species include Agalinis purpurea, Agrimonia partiflora, Apocynum cannabinum. Asclepias incarnata, Eupatorium coelestinum, E. fistu- losum, E. peifoliatum, lmpatiens capensis, Juncus spp., Mentha ^piperita, and Scirpus atrovirens. The SSG zones were present at the watershed of the SS of bodi Madison and Jackson counties where intermittent streams are located. Characteristic woody taxa include Campsis radicans, Clematis virginiana, Hy- drangea arborescens, Lindera benzoin, Sam- bucus canadensis, and Toxicodendron radi- cans. On the streambanks and gravel bars are found Bidens spp.. Boehmeria cylindrica, Eq- uisetum arvense, lmpatiens capensis. I. palli- da, Lobelia siphilitica, and Microstegium vi- mineum. Wetiand species restricted to OFR are Ci- cuta maculata, Cornus drummondii, Eleochar- is tenuis. Epilobium coloratura, Equisetum hyemale, Galium tinctorium, Juncus brachy- carpus, J. diffusissimus, J. torreyi, Leucospora multifida, Lycopus americanus, Lysimachia nummularia. Potamogeton illinoensis, Sagit- taria calycina, and Typha angustifolia. The OFR flora is made up of 47 OBL, 55 FACW, 51 FAC, 42 FACU, 10 UPL, and 9 NC species (Appendix). Red Lick Reservoir No. 2. RLR has all six wedand habitats— VOW, EM, SDF, SS, SGM, and SSG. Characteristic plants of die VOW habitat are Brasenia schreberi, Lemna minor, Najas guadalupensis, and Potomogeton nodo- sus. The EM at this reservoir lies along the east-facing bank and the southern end of the lake. Smaller areas of this habitat can also be found in the northwest corner near the dam and along the steep, west-trending bank of un- disturbed mesophstic hardwood forest. The EM zone is dominated bv Eleocharis quad- rangulata. Juncus effusus sar. solutus, Leersia oryzoides, Panicum rigidulum, Polygonum densiflorum, Scirpus cyperinus, Sparganium americanum, and Typha latifolia. The SDF habitats are located in the south end between die SS and die EM and in a few- other small sites depending on die seasonalls evaporated water levels. Characteristic species include Cyperus strigosus, Eleocharis ovata, Fimbristylis autumnalis, lmpatiens capensis, Lindernia dubia var. anagallidea. Ludwigia al- ternifolia, Penthorum sedoides, Rotala ramo- sior, and Xanthium strumarium. A small SS zone at the south end near the SSG entrance and on die disturbed west shore has Alnus glutinosa, Cornus amomum, Liquid- ambar styraciflua, Platanus occidentalis, and Salix nigra. Herbaceous plants are primarily diose of die SDF and SGM. The SGM along the west side includes Boehmeria cylindrica, Carex lurida, C. tribuloides. Equisetum ar- vense, Helianthus angustifolius, Lobelia siphil- itica, Lythrum salicaria, Mimulus alatus, Pilea pumila, and several odier species found main- ly on die SDF. The SSG habitat, located at the soudiem terminus of die reservoir, is flo- ristically different from any of die odier hab- itats. The SSG streambanks include Carpinus caroliniana, Hydrangea arborescens, Liquid- ambar styraciflua, and Sambucus canadensis; gravel bars support Boehmeria cylindrica, Carex torta, Equisetum arvense, Glyceria stri- ata, and lmpatiens capensis. Alnus glutinosa, Brasenia schreberi, Carex torta, Cornus amomum, Lemna minor, Lud- wigia decurrens, Liquidambar styraciflua. Lythrum salicaria, Panicum verrucosum, Po- lygonum densiflorum, and Sparganium amer- icanum are ssetland species found only at RLR. The RLR flora is composed of 35 OBL, 34 FACW, 32 FAC, 26 FACU, 3 UPL, and 5 NC species (Appendix). Flora ol Five Reservoirs in the Berea College Foresl Thompson and Fleming 123 DISCUSSION The five Berea College reservoirs exhibit similar stages ol hvclrosere succession com- posed of characteristic wetland species in spe- cific wetland habitats. Each reservoir varies in development of plant habitats and species composition based in part on factors of res- ervoir age, size and depth, existing vegetation contiguous to the lakes, hydrarch succession, and copper sulfate algicide treatments. The four reservoirs used for utilities water are cur- rentlv mesoligotrophie. Although Red Lick Reservoir No. 2 is the most recently formed reservoir, it has advanced to a more eutrophic stage than the older reservoirs. Some contrib- uting factors include sedimentation from ad- jacent reclaimed land, shallow depth, and a lack of direct copper sulfate treatments. The species richness of wetland and aquatic vascular plants appears to be high with 70% belonging to the OBL, FACW, and FAC cat- egories. This is a significant percentage con- sidering that the upland forested and open terrain is predominately composed of FACU and UPL species. Each resen'oir has transi- tional zones of upland forest and grassy areas bordering die wetland habitats. Many facul- tative species have taken advantage of die moisture regimens of these reservoirs. Red Lick Reservoir No. 2 has a species richness comparable to die other four reservoirs in to- tal plant species including its number of wet- land and aquatic plants. Hydrarch succession, a type of progressive secondary succession, has been induced by a number of factors at the reservoirs. The first of these factors may have been die presence of viable seed banks prior to reservoir con- struction. Once these diaspores were given the appropriate environmental conditions for ger- mination and subsequent growth, thev became established as various habitats developed. Wa- ter and wind are important dispersal mecha- nisms for propagules. The flora and fauna have shaped the floristic composition in a number of ways. Animals have aided in the distibution of viable seeds to account for a greater species richness. Waterfowl, for example, could be re- sponsible for certain wetland species that clearly were not present prior to lake forma- tion, e.g.. Aconis calamus, Brasenia schreberi, Lcmua minor. Sagittaria calycina, and Spar- gfmium americanum. The flora allows for the formation ol better-suited habitats for wetland species, through decaying of plant material, resulting in the addition ol organic material to the soil, and through increased evapotranspi- ration. allowing for areas with drier soils (Mitsch and Gosselink L993). These factors have increased the ability of these areas to support various degrees of high species rich- ness. Several wetland and aquatic species are lo- cally rare in the wetland habitats of the five reservoirs. These tax a often are restricted to a single reservoir site, i.e.. Habenaria lima. II. lacera, and H. peramoena at Lower Silver Creek Reservoir. We encountered no threat- ened, endangered, or special concern taxa for Kentucky (KSNPC 2000). We did document the presence of Polygonum densiflorum, a new Kentucky distribution record. Overall, 292 species have become established in the res- ervoirs within 10 to 77 years. ACKNOWLEDGMENTS Gratitude is extended to the Appalachian College Association, Inc., Berea, Kentucky, for financial support during summer 1998 through a Mellon Faculty/Student Research Grant awarded to Berea College. Special thanks are given to Rudv A. Gelis, a Berea College biology graduate, for field assistance and field collections from the Berea College Reservoirs. LITERATURE CITED Beat, E. O., and J. W. Thieret. 19S6. Aquatic and wetland plants of Kentucky. Kentucky Nature Preserves Coram. Sci. Techn. Ser. 5. Braun, E. L. 1950. Deciduous forests of eastern North America. Hafner Publishing Company, New York. NY. Conner. G. 19S0. Climate of Berea. Kentucky. Climatol- ogy of Kentucky series. Kentucky Climate Center, Western Kentucky Univ.. Bowling Green. KY. Cowardin, L. M., V. Carter, F. C. Golet, and E. T. LaRoe 1979. Classification of wetlands and deepwater habitats of the United States. Office of Biological Sciences. Fish and Wildlife Service, U.S.D.I. FWS OBS-79/31. Ferren. F. C... and S. J. Tonsor. 1996. Wetland classifica- tion methodology. Madrono 43:157— 178. Cleason, H. A., and A. Cronquist. 1991. Manual of vas- cular plants of northeastern United States and adjacent Canada, 2nd rd. New York Botanical Garden, Bronx, NY. Grossman, J., and t). Pittillo. 19(i2. Shrubb) and herba 124 uillliul I) I ihc Kentucky Academy of Science 65(2) ceous flora of Berea College Forest. Trans. Kentuck) Acad. Sci. 23:61-73. Hayes, R. A. 19S9. Soil survey of Jackson and Owsle) Counties, Kentucky. U.S.D.A. Soil Conservation Service and Forest Service. Washington, DC. Hoagland, B. W., and R. L. Jones. 1992. Wetland and riparian flora of the Upper Green River Basin, south- central Kentucky. Trans. Kentucky Acad. Sci. 53:141- 153. [KSXPC]. Kentucky State Nature Preserves Commission. 2000. Rare and extirpated biota of Kentuck)'. J. Ken- tucky Acad. Sci. 61:115-132. Luken, J. O., and T. N. Bezold. 2000. Species richness and wetland status of different shoreline elements at Cave Run Lake, Kentuck). Castanea 65:126-138. Meagher, W. L., and S. J. Tonsor. 1992. The checklist of the vascular flora of the Augusta Floodplain Preserve. Michigan Bot. 31:83-98. Mitsch, W. J., and J. G. Gosselink. 1993. Wedands, 2nd ed. Van Nostrand Reinhold, New York, NY. Muller, R. N., and W C. McComb. 1986. Upland forests of the Knobs Region of Kentucky. Bull. Torrey Bot. Club 113:260-280. Newton, J. H.. H. P. McDonald, D. G. Preston, A. J. Rich- ardson, and R. P. Sims. 1973. Soil survey of Madison County', Kentucky': U.S.D.A. Soil Conservation Sendee, Washington, DC. Reed, P. B., Jr. 1988. National list of plant species that occur in wedands: 1988 national summary. Biological Report 88(24). U.S.D.I., U.S. Fish and Wildlife Service, Washington, DC. Strausbaugh, P. D., and E. L. Core. 1978. Flora of West Virginia, 2nd ed. Seneca Books, Grantsville, WV. Thompson, R. L.. and E. W. J. FitzGerald, Jr. 2003. Vas- cular flora of Feltner Lake, Laurel County, Kentucky. J Kentuck) Kcad Sci, 64 75-92. Thompson, R. L., and C. A. Fleming. 2004. Vascular flora and plant communities ol the John B. Stephenson Me- morial Forest State Nature Preserve (Anglin Kails Ra- vine), Rockcastle County, Kentuck). Castanea 69:125- 138. Thompson, R. L.. and R. L. Jones. 2001. Woody plants of Rock Creek Research Natural Area and watershed up- lands. Laurel County, Kentucky Castanea 66:275—287. Trewartha, G. T, and L. H. Horn. 1980. An introduction to climate. 5th ed. McGraw-Hill Book Co., New York. NY. [USFWS]. United States Fish and Wildlife Seniee. L997. National list of vascular plant species that occur in wet- lands: 1996 national summary. National Wetlands In- ventory, U.S. Fish and Wildlife Service, Washington, DC. Wade, G. L., and R. L. Thompson. 1990. Establishment of native plant species from forest topsoil seedbanks on a borrow area in Kentucky Pages 451^160 in Proceed- ings of the 1990 Mining and Reclamation Conference and Exhibition. Vol. II. West Virginia University, Mor- gantown, WV Weir, G. W, K. Y. Lee, and P. E. Cassidv. 1971. Geologic map of die Bighill Quadrangle, east-central Kentucky GQ-900. U.S. Geological Survey (map scale 1:24,000), Washington, DC. Wood, A. J., J. M. Omemik. W. H. Martin, G. J. Pond, W. M. Andrews, S. M. Call, J. A. Comstock, and D. D. Taylor. 2002. Ecoregions of Kentucky (color poster with map, descriptive text, summary tables, and photo- graphs). U.S. Geological Survey (map scale 1: 1,000,000), Reston, VA. Flora of Five Reservoirs in the Berea College Forest — Thompson and Fleming 125 Appendix. Vascular plants of the live- Berea College Reservoirs, Madison and Jackson counties K. ntucky. Relative ubundj Taxon Nations] Wetland Category [wetland habitat] hi CBR lilH Equisetophyta Equisetaceae Equisetum arvense L. FAC. [SSG] E hyemale L. FACW. [SS] Lycopodiophyta Selaginellaceae Selaginella apoda (L.) Fern. FACW. [SSG] Polypodiophyta Aspleniaceae Athyriumfilix-femina (L.) Roth. FAC. [SSG] A. pyenocarpon (Spreng.) Tidest. FAC. [SSG] A. thelypterioides Michx.) Desv. FAC. [SSG] Polystichum acrostichoides (Michx.) Scliott. FACU-. [SSG] Thelypteris hexagonoptera (Michx.) Weatherbv. FAC. [SSG] Onocleaceae Onoclca sensibilis L. FACW. [SCM] Ophioglossaceae Bolrychium dissectum Spreng. FAC. [SDF] Pinophyta Taxodiaceae Taxodium distichum (L.) Rich. OBL. [SDF] Magnoliophyta Aceraceae Acer negttndo L. FAC + . [SDF] A. nibnim L. FAC. [SSG] A. saccharinum L. FACW [SS] Acoraceae Acorus calamus L. OBL. [EM] Ahsmataceae Alisma subcordatum Raf. OBL. [EM] Sagittaria australis (J.G. Smith) Small. OBL. [EM] S. calycina Engelm. OBL. [EM] Anacardiaceae RJms copallina L. FACU-. [SSG] fi. glabra L. NC. [SGM] Toxicodendron radicans (L.) Kuntze. FAC. [SDF] Apiaceae Cicuta maculate L. OBL. [SGM] Cryptotaenia canadensis (L.) DC. FAC. [SGM] *Daucus carota L. NC. [SGM] Osmorhiza claytonii (Michx.) Clarke. FACU-. [SGM] Sanicula canadensis L. UPL. [SGM] Apocynaceae Apocynum cannabinum L. FACU. [SGM] Asclepiadaceae Ampelamus albidus (Nutt.) Britt. FAC. [SDF] Asclepias incarnaia L. OBL. [SDF] \ syriaca L. FACU-. [SGM] A. tuberosa L. NC. [SGM] A. nitidis Walt. NC. [SGM] Asteraceae *AchUlea millefolium L. FACU. [SGM] Ambrosia artemisiifolia L. FACU. [SGM] A. trifida L. FAC. [SGM] Aster cordifolius L. NC. [SSG] A. dumosus L. FAC. [SGM] A. divaricatvs L. NC. [SSG] A lateriflorus (L.) Britt. FACW-. [SDF] A. ontarionis Wieg. FAG. [SD1 O O I n R I 1 1 O 1 0 1 R R I I o I () R R I I o F I R o I R o I I F O 1 o O F O I — I R I — I o I O 1 R — I R — R o I 1 — O o o F I I o 1 1 F 1 I 1 1 1 R — R — R — R 1 R O I 0 O I 0 o — O — 0 o 0 () 0 I 1 1 [ ! 1 I 1 1 126 [ourna] oJ the Kentuck) Academy oi Science 65(2 Appendix, ( lontinued. Taxon, National Wetland Categoiy. [wetland liabital .A. pOosus Willd. UPL. [SGM] A. prenanthoides Muhl. FAC. [SDF] A. puniceus L. OBL. [SDF] Btdens rcnnm L. OBL. [SDF] B. frondosa L. FACVV. [SGM] B.polylepis S. F. Blake. FACW. [SGM] ^Chrysanthemum leucanthemum L. UPL. [SGM] Comjza canadensis (L.) Cronq. UPL. [SGM] *Eclipta prostrata (L.) L. FAC. [SDF] Elephantopus carolinianus Willd. FACU. [SGM] Erechtites hieracifolia (L.) Raf. FACU. [SGM] Eupatorium coelestinum L. FAC. [SGM] E. fistulosum Barratt. FACW. [SDF] E. purpureum L. FAC. [SGM] E. perfoliatum L. FACW+. [SGM] E. rotundifolium L. FAC-. [SGM] E. serotinum Michx. FAC-. [SGM] Euthamia graminifolia (L.) Nutt. FAC. [SGM] Helenium flexuosum Raf. FAC—. [SDF] Helianthus angustifolius L. FACW. [SDF] Helianthus microcephalus T. & G. NC. [SGM] 7r<7 annua L. FAC. [SGM] Lactuca canadensis L. FACU — . [SGM] P/uc/iea camphorata (L.) DC. FACW. [SDF] Polynmia uvedalia L. FAC. [SGM] Prenanthes altissima L. FACU-. [SGM] Pyrrhopappus carolinianus (Walt.) DC. NC. [SGM] Rudbeckia fulgida Ait. FAC. [SDF] R hirta L. FACU. [SGM] R triloba L. FACU. [SGM] Silphium trifoliatum L. FAC. [SGM] Solidago caesia L. FACU. [SDF] S. canadensis L. FACU. [SGM] S. flexicaulis L. FACU. [SDF] S. glgantea Ait. FACW. [SDF] S. rugosa P. Mill. FAC. [SGM] *Sonchus asper (L.) Hill. FAC. [SGM] Verbesina alternifolia (L.) Britt. FAC. [SGM] Vernonia gigantea (Walt.) Trel. FAC. [SGM] Xanthium struniarium L. FAC. [SDF] Balsaminaceae Impatiens capensis Meerb. FACW. [SGM] I pallida Nutt. FACW. [SSG] Betulaceae *Alnus glutinosa (L.) Gaertn. FACW-. [SS] Betula nigra L. FACW. [SS] Carpinus caroliniana Walt. FAC. [SS] Corylus americana Walt. FACU—. [SWG] Bignoniaceae Campsis radicans (L.) Seem. FAC. [SSG] Brassieaceae *Cardamine hirsuta L. FACU. [SGM] Lepidium virginicum L. FACU—. [SGM] Rorippa palustris (L.) Bess. OBL. [EM] Cabombaceae Brasenia schreberi J. F. Gmel. OBL. [VOW] Campanulaceae Campanula americana L. FACU. [SGM] Lobelia cardinalis L. FACW+. [SGM] L. inflata L. FACU. [SDF] R — — 0 I I — 1 1 o o 0 1 0 I o I 0 1 R — [ o I — — — R R R — o 1 O I I F R O 1 O — O O I I F O I o O O — o o O I — — o R O I o O R R I R R I I R — — I — — I R — — R I R — — I — I — O I I O I — F — O I O o O I — — — R O — — o — — R o O — I — I — I o O F I F A F I I I I R — I I I I I R O I I I o — — F I I — I o I Flora of Five Reservoirs in the Berea College Forest — Thompson and Fleming 127 Vppendix. < Continued. Taxon. National Wetland Categoi) [wetland I 1 M use CBR OFR Rl.K 1 1 1 I 0 C) R 1 O I n F F () 1 0 1 R () 1 1 0 O O 0 1 1 I 1 1 1 I I o 1 I I 1 o R 1 O I R — I o I 1 R I I I F I R I I o 1 R I F O R — O o I 1 F I o O o I I O o R O o I F F I F F F O O o o — F F — O — O I 1 I F R O F F F O I R — I O R — F 1 F F F 1 1 O — O O R O I — R R — 1 I O 1 — — R R — O F O F O — — — 1 O L puberula Michx. FACW-. [SDF] L, syphilitica I.. FACW+. [SGM L. spicata Lam. FAC — . [SGM] Triodanis perfoliata L Nieuwl. FAC. siai Caprifoliaceae *Lonicera japonica Thunb. FAC—. [SGM Sambucus canadensis L. FACW. [SWG Viburnum rufidulum Raf. UPL. [SDF] Clusiaceae Hypericum mulilum L. FACW. [SDF] H punctatum Lam. FAC-. [SDF] Commelinaceae *Commelina communis L. FAC—. [SGM! Convolvulaceae Calystegia sepium L. R. Br. FAC-. [SGM] *lpomoea laatnosa L. FACW. [SDF] 7. pandurata (L.) G.Mey. FACC. [SGM] Cornaceae Cornus amomum P. Mill. FACW. [SS] C. drummondii C. A. Mey. FAC. [SS] Cuscutaceae Cuscuta gronovii Wild. NC. [EM] C. indecora Choisv. NC. [SDF] C. pentugona Engelm. NC. [SGM] C\peraceae ' Carexfrankii Kunth. OBL. [EM] C. hirsutella Mack. FACC. [SDF] C. lurida Wahl. OBL. [EM] C. squarrosa L. FACW. [EM] C. stipata Wild. OBL. [EM] C. torta F. Booth. FACW. [SSG] C. tribuloides Wahl. FACW+. [EM C. mdpinoidea Michx. OBL. [EM] Cyperus breiifolioides Thieret 6c Delahoussave FACW. [SGM] C. flavescens L. OBL. [SDF] C. strigostis L. FACW. [EM] Eleocharis ovata (Roth) R. & S. OBL. [SDF] £. palustris (L.) R. & S. OBL. [EM] E quadrangulata Michx.' R. & S. OBL. [EM] E. tenuis (Willd.) Schult. FACW+. [SDF] Fimbristylis autumnalis L. R 6c S. FACW+. [SDF] Rlujnchospora capitellata (Michx.) Valil. OBL. [EM] Scirpus atrovirens Muhl. OBL. [SDF] S. cyperinus (L.) Kunth. FACW+. [SDF] S. pendulus Muhl. OBL. [EM] S. pohjphtjllus Vahl. OBL. [EM] S. purshianus Fern. OBL. [EM] S. calidus Vahl. OBL. [EM] Scleria triglomerata Michx. FAC. [SGM] Dioscoreaceae *Dioscorea batatas Dene. NC. [SSG] Euphorbiaceae Acahjpha rhomboidea Raf. FACU— . [SDF] X i-'irginica L. FACC-. [SDF] Euphorbia maculata L. FACC — . [SDF' £. nutans Lagasca. FACU—. [SDF] PhyUanthus carvlinicnsis -Walt. FAC- SDK Fabaeeae Amphicarpaea bracteata I.. Fern. FAC. [S(.\l Apios americana Metlik. FACW. .EMi L28 journal nl the Kentuck) Acadi f Science 65(2) Appendix. ( Continued. Relative abundances Toxon. \.[i i! Wetland * ategory. [wetland habitat] Chamaecrista fasciculata (Michx.) Greene. FACU. [SGM] Desmanthus ittinoensis (Michx.) MacMill. FAG [SGM] Desmodium paniculatum (L.) DC. UPL. [SGM] *Lespedeza cuneata (Duni Cours.) Don. FACU — . [SDF] L. intermedia (S. Wats.) Britt. NC. [SGM] *L. stipulacea Maxim. FACU. [SDF] *L. striata (Thunb.) H. & A. FACU. [SDF] *Melihtus alba Desr. FACU-. [SGM] Strophostyles umbellata (Muhl.) Britt. FACU-. [SGM] *Trifolium pratense L. FACU-. [SGM] Gentianaceae Sabatia annularis (L.) Pursh. FAC + . [SDF] Hamamelidaceae Liquidambar styraciflua L. FAC. [SS] Hydrangeaceae Hydrangea arborescens L. FACU. [SSGF] Juncaceae Juncus acuminatum Michx. OBL. [EM] /. biflorus Ell. FACW. [SGM] /. brachycarpus Engelm. FACW. [SGM] /. diffusissimus Buckl. FACW. [SGM] /. dudleiji Wieg. FAC-. [SGM] J. effusus L. var. solutus Fern. & Wieg. OBL. [EM] /. marginatus Rostk. FACW. [SDF] /. tenuis Willd. FAC-. [SGM] /. torreyi Cov. FACW. [SGM] Lamiaceae Collinsonia canadensis L. FAC+. [SGM] Lycopus americanus Muhl. OBL. [EM] L. virginicus L. OBL. [SDF] "Mentha ^.piperita L. FACW+. [SGM] Physostegia virginiana (L.) Benth. FAC + . [SSG] "Prunella vulgaris L. FACU. [SGM] Pycnanthemum tenuifolium Schrad. FACW. [SDF] Scutellaria elliptica Muhl. NC. [SDF] Scutellaria lateriflora L. FACW+. [SDF] Stachys nuttatlii Schuttw. FAC. [SGM] S. tenaifolia Willd. FACW+. [SDF] Teucrium canadense L. FACW—. [SDF] Trichostema dichotomum L. NC. [SGM] Lauraceae Lindera benzoin (L.) Blume. FACW-. [WM] Lemnaceae *Lemna minor L. OBL. [VOW] Liliaceae "Allium vineale L. FACU-. [SGM] Lilium canadense L. FAC + . [SGM] Linaceae Linum medium (Planch.) Britt. FACU. [SDF] L. striatum Walt. FACW. [SDF] Lythraceae Ammannia coccinea Rottb. OBL. [SDF] *Lythrum salicaria L. FACW+. [SGM] Rotala ramosior (L.) Koehne. OBL. [SDF] Magnoliaceae Liriodendron tulipifera L. FACU. [SS] Melastomataceae Rliexia virginica L. OBL. [SDF] Molluginaceae *Mollugo verticillata L. FAC. [SDF] — () — F 1 0 — — 0 F — 0 1 0 1 O — — 0 — — F () 1 1 — I 1 1 — F — — O I — — — — I I o o O I I o o o O — — R o 1 o — — — — R F o F o F — I I o I I o I F I O I I R I — — — O — o o I F 1 — I — O — — — — R — I o R O I o I I F — O R O O F I R R I — R 0 I I — 1 I I R — — — I I O F F o — — — R — R R I I — 0 F I O I O O — O — o — _ o Flora <>l Five Reservoirs in the Berea College Foresl Thompson and Fh mine. 129 Appendix. < lontinued, Tilwhi. National Wetland Category [wetland habitat] lie m use LSC dill OFB I'.l.li ( 1 ( ) F F F F I A 1 A 1 F o F I R () o 1 R F o F H F F I R R O R R — o I 0 I 0 O 1 1 o O 1 I 0 o o I R — o o — R I o o R I o I I o I I — I I I I I I 0 I R I I o o F O o o o F F o o F O — o F A F A O F F , i F O I F F F 0 — O I I — F F F F I F F 1 O F O F O F 1 — I o F 1 1 o O o F R o — 1 I — 1 R O I o — o Najadaceae *Najas minor AH. OBL. [VOW] N. guadelupensis (Sprcng.) Magnus. OBL. [VOW] Fraxinus pennsylvanica Marsh. FACW. |SS] Onagraceae Circaea lutetiana L. FACU. [SGM] Epibbium colomtum Bi.-liler. FACW+. [SDF] Ludwigia alternifolia L. FACW+. [SDF] /.. decurrens Walt. OBL. [EM] L. palustris (L.) Ell. OBL. [SDF] Orchidaceae Habenariaflava (L.) R. Br. FACW. [SGM] II lacera (Michx.) Loud. FACW. [SGM] H. peramoena A. Gray. FACW. [SGM] Spiranthes cernua (L.) Rich. FACW. [SDF] S. lacera (Raf.) Raf. FACU-. [SGM] PJantaginaceae Plantago rugelii Dene. FACU. [SGM] Platanaceae Platamis orcidentalis L. FACW-. [SS] Poaceae *Agrostis gigantea Roth. FACW. [SDF] A. perennans (Walt.) Tuckerm. FACU. [SGM] Andropogon oirginicus L. FACU. [SGM] Brachyelytrum erectum (Schreb.) Beauv. NC. [SGM] Cinna arundinacea L. FACW+. [SDF] *Digitaria ischaemum (Schreb.) Muhl. UPL. [SDF] *D. sanguinalis (L.) Scop. FACU-. [SDF] *Echinochloa crus-gaUi (L.) Beauv. FACU. [SDF] E. muricata (Beauv.) Fern. FACW+. [SDF] *Eleusine indica (L.) Gaertn. FACU-. [SDF] Elymus htjstrix L. UPL. [SSG] E. oirginicus L. FACW-. [SDF] *Efagrostis pectinacea (Michx.) Nees. FAC. [SDF] *E. spectabilis (Pursh) Steudel. UPL. [SGM] *Festuca elatior L. FACU-. [SGM] Glyceria striata (Lam.) Hitehc. OBL. [EM] *Holcus lanatus L. FACU. [SGM] Leersia onjzoides (L.) Swartz. OBL. [EM] L. virginica Willd. FACW. [SDF] *Microstegium oimineum (Trin.) Camus. FAC. [SSG] Muhlenbergia frondosa (Poir.) Fern. FAC. [SSG] Panicum anceps Michx. FAC. [SGM] P. clandestinum L. FAC + . [SDF] P. dichotomiflorum Michx. FACW-. [SDF] P. dichotomum L. FAC. [SDF] P. flexile (Gattinger) Scribn. FACU. [SDF] P. lanuginosum Ell. FAC. [SDF] P. polyanthes Schult. FAC. [SDF] P. rigidulum Nees. FACW+. [EM] P. verrucosum Muhl. FACW. [SGM] Paspalum laeve Michx. FAC + . [SGM] *Setaria faberi Herrm. UPL. [SGM] S. geniculata (Lam.) P. Beauv. FAC. [SGM] *Sorghum halepense (L.) Pers. FACU. [SDF] Tridens flams (L.) A. Hitehc. FACU. [SDF] Polygalaceae Tolygala ambigua Nutt. UPL. [SGM] P. sanguinea L. FACU. [SGM] 130 journal ol the Kentucky Academy ol Science 65(2) Appendix. Continued. Relative abundam T.l\l Boone County — Bryant Galbraith, ami Held 133 H«w U .. t\ \G ^J^7'«^\-f. Figure 1. A glacial map of Boone County, Kentucky, and the Greater Cincinnati region showing the extent of drift deposits (after Ray 1974). increasing human population and subsequent changes in land use. METHODS AND MATERIALS In general, trees at all sites were sampled in 0.04 ha circular plots spaced at 30 m inter- vals along line transects throughout each for- est. Two stands were sampled using plotless methods, and the prairie was sampled in 1 m X 1 m plots (Bryant 1981b). We sampled 17 mature forests plus one prairie site from across Boone County; however, one of the for- est sites was located in adjacent Kenton Coun- ty (Figure 2). Onlv trees ^10 cm diameter breast height (dbh) were measured; however, in two stands die minimum dbh was &S.9 cm. Although shrubs, seedlings and saplings, and herbs were sampled in most stands, only overstory was considered in the forest analysis for this paper. These data were analyzed to relative frequen- cy (RF), relative density (RD), and relative dominance (RDo), which were then summed to generate an importance value (IV) for each species (Curtis and Mcintosh 1951). Densitv (trees/ha), total basal area (m2/ha), and species diversity (H') were calculated for each forest. Grasses and forbs in the prairie were analyzed to frequency onlv (Brvant 1981b). Classification of stands was developed using an unweighted pair-group mean-average (UPGMA) cluster analysis with percent simi- larity (Kovach 199S). A cutoff value of 60% similarity was used to create the initial cluster groupings (Hinkle 197S). This analysis was performed on untransformed importance val- ue data, which included all species. Stand re- lationships were summarized using the Bray- Curtis ordination procedure (McCune and Medford 1999). RESULTS Fortv-four tree species were recorded for Boone County, 38 from our samples plus 6 from Keith (1968). Tree density, basal area, tree species diversity, and tree species richness 134 Journal ot the Kentucky Academy <>l Science 65(2) Table 1. The- percentage '' "I land in Boone County, Kentucky with residual and transported soils. Tile total percentage does not equal 100 since only soil types for sampling sites are listed. Also the range of slope for each soil type is included. Slope Residual soils Eden siltv clay loam Cynthiana flaggy' silty clay loam Faywood siltv clay loam Total' 19.0 8.8 13.9 41.7 12-35 12-50 2-20 Transported soils Glacial (loess over glacial till) Rossmoyne silt loam (fragipan) Jessup silt loam (no fragipan) 23.1 14.5 0-12 2-20 Loess (loess over residuum) Nicholson silt loam Alluvial land, steep Lakin loamy- fine sand 7.9 1.4 0.6 0-12 0-12 Alluvium Lindside silt loam Nolin silt loam Huntington silt loam Total 0.9 1.1 1.2 50.7 0-4 0-4 0-4 for each stand are shown in Table 2. Based on cluster analysis, we were able to recognize three vegetation types: (1) mixed mesophvtic (glacial), (2) oak-hickorv, and (3) oak-ash-ma- ple (Figure 3). All five of the mixed meso- phvtic (glacial) stands clustered together at >60% similarity as did two oak-hickorv stands and two oak-ash-maple stands. Other vegeta- tion types apparently were represented bv only one stand each and did not show cluster- ing. In an attempt to determine vegetation-en- vironmental relations, all stands were ordinat- ed. Axis 1 of the ordination appeared to be a moisture gradient with the prairie at the drier end and the floodplain forest at the wetter end (Figure 4). The majority of stands tended to group near the middle of the moisture gradi- ent, perhaps reflecting the mesic nature of die county. Axis 2 was either a topographic or soil gradient, but may be a combination of die two. The beech forest was on a flat upland site with Jessup soil, die mixed mesophytic (steep alluvial) site was in a ravine widi sandy soils of Illinoian or glacioflmiatile origin (Ray 1974), and the beech-maple site was gendy rolling with Rossmoyne soil, which consists of Wis- consin loess over glacial till (Weisenberger et al. 1973). All mixed mesophytic ( glacial i sites were in highly dissected areas ot Kansan out- wash and Jessup soils. The oak-hickory sites were on gentle to moderate slopes with Fay- wood and/or Eden silt loams; the oak-ash-ma- ple sites were on steep slopes with Cynthiana silt loams. The latter three soil types are re- sidual. DESCRIPTION OF VEGETATION Mixed mesophvtic forests were of two t\pes, those on Kansan glacial deposits and others on steep alluvium. The mixed meso- phytic glacial sites (BC, BN, BP, MU, YP; codes for sites are explained in Figure 2) xvere on highly dissected outwash deposits of Kan- san age. Seeps and springs were characteristic of these sites. Dominant tree species included sugar maple (Acer sacchamm), white ash (Fraxinus americana), beech (Fagus grandi- folia), bassvvood (Tilia americana), northern red oak (Quercus rubra), bitternut hickory (Carya cordifonnis) , and tulip poplar (Liriod- endron tulipifera). Tree species diversity (H') ranged from 2.6 to 3.5 (Table 2) and these sites are floristically rich (Bryant 1978). Tree density ranged from 222 to 499 trees/ha, and basal areas fit die >25 m2/ha proposed bv Martin (1992) for mature mixed mesophvtic forests. Mixed mesophvtic (steep alluvium) forests are found in ravines (SC) with steep alluvial soils. These sandy soils are found within die narrow band of Illinoian glacial deposits near die Ohio River in the western portion of the county (Figure 1). Tulip poplar was the dom- inant species at this site and also at a sandv hummock site (HS); however, vegetation at that site tended to ordinate more closely to beech-maple than to the mixed mesophvtic (glacial) stands. Oak-hickory forests (DU, WS) were associ- ated with the droughty residual Fawvood or Eden soils, although other soils, e.g., Nichol- son, were of minor importance. Those stands were located on side slopes and narroxv ridges, especially- widiin the Eden Shale Belt portion of Boone County. White oak (Quercus alba), northern red oak, and shagbark hickory (Car- ya ovata) were canopy dominants. On such sites sugar maple was primarily a subcanopy member (Bryant 1981a). Odier stands (LW, MM, DW) showed similarities to oak-hickory, Vegetation ol Boone County — Bryant, Galbraith, and Held 135 Figure 2. Location of sampling sites in Boone County, Kentucky. Site abbreviations: BC, Boone County Cliffs Nature Preserve; BM, Beech-Maple; BN, Boone County Cliffs Addition; BP. Bald Point; DU, Durrs Woods; DW, Dinsmore Woods; EB, East Bend; EC, Elijah Creek; GC, Gunpowder Creek; HS, Hummock Site; KP, Kansan Prairie; LW, Luebber's Woods; ML, Middle Creek Floodplain; MM, Middle Creek Middle Slope; Ml!, Middle Creek Upper slope; SC, Steep Creek; WS, Woodland-Scott (located in adjacent Kenton Count)); and VP, Young Property. Open circles represent Keidi's (1968) sample sites; closed circles show location of some towns in Boone Countv. but other oak species, not white oak, were of greater importance. Oak-ash-maple communities (EC, GC. DW) occupied moderate to steep slopes, es- pecially where erosion had removed much or all of the former till deposits. Cvnthiana silt loam was the principal soil type and the forest dominants were oaks (Quercus spp.), white ash, and sugar maple. Because of composi- tional similarities, it was often difficult to dis- tinguish oak-hickorv and oak-ash-maple stands. Beech-maple (BM) forests yvere most prominent on the level to gently rolling up- lands with Rossmovne soils. These loessial soils are underlain by a fragipan that may hin- der drainage. Beech (EB) forests were also found on gentlv rolling uplands, but there Jes- sup soil is most prominent. Both lorest types had low tree densities, but high basal areas (Table 2). Floodplain forests were dominated by box- elder (Acer negundo), cottonwood [Papains deltoids), and silver maple (.4. saccharinum . These forests are located on wide creek banks and Ohio [liver backwater sites. The alluvial 36 journal ol the Kentuck) Academ) of Science 65(2) KP ML BM SC | LW __ M 1 H L H I w 1 BP BC -20 20 40 60 80 100 Percent Similarity Figure 3. Dendrogram based on cluster analysis, showing the relationship of the sites sampled in Boone County, Kentucky. CM tn x < GC DW DU " EC WS" LW MM- * BC YP HS BP BH „MU KP BM SC EB ML Axis 1 Figure 4. Ordination of the 18 sites sampled in Boone County. Axis 1 is a moisture gradient; a\is 2, a topographic/ soils gradient. Vegetation ol Boone C -oiint\'— Bn/aitl. C.alliniilli. rind Held 13' Table 2. Tree density (trees/ha), basal area (m2/ha), spe cies diversity (H'l. and the number of tree species for forests .mil other environs in Boone County. Keiilnrkv Site abbreviations as in Figure 2. Smith's richness (number «»! tree sjm tics i Forest site ha m ha II' BC 499 27.S 3.2 25 BN 390 28.3 2.6 IS BP 410 38.4 3.5 20 MU 262 24.7 2.1 12 YP 222 27.1 3.2 23 SC 302 46.0 3.7 20 IIS 296 36.2 2.8 16 BM 233 32.6 3.0 18 EB 126 35.4 2.0 8 ML 538 26.7 0.9 9 DU 470 23.1 2.3 17 WS 343 31.4 2.1 13 I.W 347 26.6 1.4 11 MM 314 21.5 3.0 17 DW 334 28.1 3.2 22 EC 328 22.4 3.0 14 GC 457 29.6 2.8 16 KP 0 0 0 0 soil complex included Lindside, Nolin, and Huntington silt loams. Species diversity at the wet end of the moisture gradient was lower than for mesic and drv-mesic sites. Prairie communities (KP) were rare; occur- ring as small isolated patches on exposed gla- cial outcrops or upland sand deposits (Bryant 1981b). Little blustem (Schizachyrium scopar- ium), side-oats grama (Bouteloua curtipendu- la), and Indian grass (Sorglwstmm nutans) were characteristic of these drier sites al- though a number of forbs were relatively abundant. Seedlings of eastern redcedar (Jun- iperus virginiona) appeared to be slowly in- vading the prairie. DISCUSSION There have been few recent reports or de- scriptions of countvwide (e.g., Campbell and Grubbs [1992] for Hopkins County) or region- al vegetation (e.g., Bryant and Held [2001] for the Jackson Purchase Region) in Kentucky. However, Bryant and Held (2004) recently de- tailed the vegetation-environment patterns in Hamilton County, Ohio. Geologically, Boone and Hamilton counties have experienced sim- ilar glacial events; phytogeographicallv, these two counties are located in the northern por- tion of Braun's (1950) Western Mesophvtic Forest Region but near the junction of her Mixed Mesophytic, Oak-Hickory, and Beech- Maple Fores! Regions. The structure and composition of vegeta- tion are determined l>\ a nber of factors including past disturbances, successional pro- cesses, and individualistic responses of species to environmental constraints (Cole and Ware 1997). A number of. species respond similarih to environmental gradients and thus sort oul as communities or vegetation types. The beech-maple and beech forests that we identified may represent remnants of forest types that formerly were more widel) distrib- uted on Rossmoyne soils in Boone and adja- cent Kenton counties, Kentuck) (Keith L968). Beech-maple forests were reported on soils of mixed glacial origins in Hamilton County, Ohio (Bryant 1987; Bryant and Held 2004) and were found on Illinoian deposits well south of the Wisconsin glacial border in In- diana (Lindsey et al. 1965). Braun (1950) and Vankat et al. (1975) considered the terminus of the Wisconsin advance in southern Ohio to be the southern extent of beech-maple forests. Bryant and Held (2004) reported two hpes of mixed mesophytic forests in Hamilton County, one with tulip poplar and one with- out. We also found two tvpes of mixed meso- phytic forests in Boone Count)' — one on high- ly dissected Kansan outwash deposits (Bryant 1978) and one on steep alluvium. Although Keith (1968) considered tulip poplar to be scarce in Boone County, it was common in both mesophvtic tvpes and was a dominant on the steep alluvium. In soutiiern Ohio, Forsvth (1970) noted diat mixed mesophvtic forests containing buckeye, beech, white basswood. and tulip tree appeared to correlate with die occurrence of Kansan drift. A great variety of moisture conditions exists in these deep, steep-sided valleys (Forsvth 1970) and sup- ports a mixed mesophvtic association. We found the floodplain forests in Boone Count)' to be eompositionallv similar to those in Hamilton Countv. Boxelder, Cottonwood, and silver maple, along with green ash (F. pennsylvanica) and various willows {Salix spp.), vary in importance in different parts of die floodplains. Oak-hickory forests were lound primarily on steep slopes and ridges with droughty residual soils. Keith (1968) speculated that white oak had been more important in the past than at 138 il of tlic Kentucky Academy ol Science 65(2) present. In the Eden Shale Belt, white o;ik is generally more important in stands where dis- turbance has been minimal (Bryant 1981a), but its importance was reduced on previously logged sites. This may support Keith's (1968) speculation; however, slope aspect may influ- ence the occurrence of white oak. On less ex- posed aspects, more mesic species are of greater importance. Oak-ash-maple stands were most commonly found on steep slopes in those areas of the count)' where erosion had removed the former glacial cover. In Hamilton Count}-, Ohio, Bry- ant and Held (2004) referred to this forest type as western mesophvtic after Gordons (1966, 1969) recognition of western meso- phvtic forests in southern Ohio. Prairies, although extremely small in areal extent, add an important component to the county's vegetational diversity. These prairie remnants have been maintained by a complex of factors including periodic disturbances (Bryant 1981b). SUMMARY The broad vegetational development shown for Boone County might not be expected on a local level (i.e., one count}7); however, geo- logic and edaphic diversity7 underlie and pro- mote biological diversity here. In adjacent Hamilton County, Ohio, Bryant and Held (2004) found forest types to sort out along moisture and topographic gradients. We found a similar pattern in Boone County but perhaps widi a stronger influence from soils, especially in relation to glacial and postglacial history. A mosaic of unlike climaxes is characteristic of the Western Mesophvtic Forest Region (Braun 1950); we, too, found a mosaic of veg- etation types. Braun (1950) considered the Western Mesophvtic Forest Region to repre- sent a tension zone where the compensating effects of local environments permit unlike cli- maxes to exist close to one another. We sup- port that tension zone interpretation for Boone County, especially considering its geo- graphic location and glacial-edaphic history. LITERATURE CITED Braun, E. L. 1950. Deciduous forests of eastern North America. The Blakiston Co.. Philadelphia, PA. Bryant. W. S. 1978. Vegetation of the Boone County- Cliffs Nature Preserve, a forest on a Kansan outwash deposit in northern Kentucky. Trans, Kentucky Acad. Sci, 39: 12-22. Bryant, W. S. 1981a. Oak-hickor) forest of the Eden Shale Belt: a preliminary report. Trans. Kentucky Acad Si i 42:41-15. Bryant, W S. L9811). Prairies on Kansan outwash deposits in northern Kentucky. Ohio Biol. Surv. Biol. Notes 15 8,8-91. Bryant. W. S. 1987. .Structure and composition of the old- growth forests ol Hamilton County, Ohio and environs. Pages 317-324 in R. L. Hay, F. W. Woods, and II. DeSelm (eds). Proc. Sixth Centra] Hardwoods Forest Conference. University of Tennessee, Knoxville, TN. Bryant, W. S., and M. E. Held. 2001. An ordination of the plant communities of the Jackson Purchase flegion of Kentucky*. Pages 11-18 in E. W. Chester and A. F. Scott (eds). Proc. Ninth symposium on the natural history of Lower Tennessee and Cumberland River vallevs. The Center for Field Biology, Austin Peay- State University. Clarksville, TN. Bryant, W. S., and M. E. Held. 2004. Forest vegetation in Hamilton County, Ohio: a cluster analysis and ordi- nation study. Pages 312-321 in Proc. 14th Central Hardwood Forest Conference. GTR-NE-316. Campbell, J. J. N., and J. Grubbs. 1992. Natural plant communities of Hopkins Counts, Kentucky. Trans. Ken- tucky Acad. Sci. 53:29-38. Cole, A. M., and S. A. Ware. 1997. Forest vegetation, edaphic factors, and successional directions in the cen- tral Piedmont of Virginia. Castanea 62:100-111. Curtis, J. X, and R. P. Mcintosh. 1951. An upland forest continuum in the prairie-forest border region of Wis- consin. Ecology 32:476-496. Elam, A. B„ Jr. 1973. Climate. Pages 63-64 in B. C. Wei- senberger, C. W. Dowell. T R. Leathers, H. B. Odor, and A. J. Richardson (eds). Soil survey of Boone, Camp- bell, and Kenton counties, Kentucky. U.S.D.A. Soil Conservation Service. Government Printing Office, Washington, D.C. [ERMC]. Environmental Resource Management Center. 2002. Boone County Forest Quality Assessment: an ecological evaluation, prioritization, and mapping. RPt: 1090. Forsvth, J. L. 1970. A geologist looks at die natural veg- etation of Ohio. Ohio J. Sci. 70:180-191. Held, M. E., and J. E. Winstead. 1976. Structure and composition of a climax forest system in Boone County, Kentucky. Trans. Kentucky Acad. Sci. 37:57-67. Hinkle, C. R. 1978. The relationship of forest communi- ties and selected species to edaphic and topographic factors on the Cumberland Plateau of Tennessee. Ph.D. Dissertation. University of Tennessee, Knoxville, TN. Gordon, R. B. 1966. Natural vegetation of Ohio, at die time of the earliest surveys [map]. Ohio Biological Sur- vey, Columbus, OH. Gordon, R. B. 1969. The natural vegetation of Ohio in pioneer davs. Ohio Biol. Bull. n.s. 3(2). Keith, J. R. 1968. Vegetation of the Pleistocene Drift Re- Vegetation of Boone County — Bn/aul. Galbraith, anil Held 139 gion, northern Kentucky Trans. Kentucky Acad. Sci. 29: 1O-20. Kovach, W. L. 1998. MVSP— a multivariate statistical package for Windows. Version '>.(). Kovach Computing Services. Pentraeth, Wales. U.K. Lindsey. A. A.. W. B. Crankshaw, and S. A. Qadir. 1965. Soil relations and distribution map ol vegetation ol piv- settlement Indiana. Bot. Gaz. 126:15.5-163. Martin. W. II. 1992. Characteristics of old-growth mixed mesophytic forest. Nat. Areas J. 12:127-135. McCune, B., and M J Medford. 1999. PC-ORD for Win- dows (Version 4.17): multivariate analysis ol ecological data. MJM Software. Gleneden Beach. OR. Middleton. A. R.. W. R. Jillson. F. T. McFarland. and W. V Vnderson 1926 Kentucky. Pages 349 154 in \ I Shelford (ed Naturalist's guide to the vmericas The William', ami Wilkins Co Baltimore, Ml) Ray, L. I.. 1974. Geomorpholog) and quaternary geolog) ol the glaciated Ohio River Valle) — a reconnaissance stud). U.S. Geol. Sun. Professional Paper S2b. Vankat, J. I... W II. Blackwell, and W. E. Hopkins 1975 The dynamics of Hueston Woods and a review "I the question of the successional status of the southern beech-maple forest. Castanea 40:290-308. Weisenberger, If C, E. W. Dowell, T. R. Leathers. II. B. Odor, and A. J. Richardson. 1973. Soil surve) of B( Camphell and Kenton counties. Kentucky. U.S.D.A. Soil Conservation Sen ice Government I'rintiiig Office Washington, DC. J. Ky. Acad. Sci. 6S(2):140-153. 2004. Scientists of Kentucky Christopher Columbus Graham: Kentucky Man of Science James Duval] Boone County Public Library, Scheben Branch, 8899 U.S. 42. Union, Kentuck) 41091 Christopher Columbus Graham (1784- 1885) was bom in Kentucky before it became a state. He was considered one of the most interesting and useful citizens in the state long before die time of his death at the age of 100. Though he was a man concerned in public af- fairs and a self-made man of wealth, he never held a public office. He was a traveler, writer, archaeologist, medical doctor, civic leader, and philanthropist.1 In die larger historical picture his scientific contributions must be considered negligible, but he participated in the scientific life of die time and might be considered char- acteristic, as there were very few men then who could be considered professional scien- tists. As a woodsman Graham had few equals. When a young man he made about 20 flatboat trips down the Mississippi. He was on die wa- ter near New Madrid, Missouri, during the great earthquake of December 1811. He de- veloped such facility with die rifle that he eventually came to be regarded as die best marksman in the world, and he could shoot with unaided vision to die age of 100. ' The skills he gained at this period of his fife were doubtless of value when he joined a company of infantry during the War of 1812. - He was wounded, captured, then exchanged, re-enter- ing active service. Later, at Fort Maiden, Can- ada, he was captured by Indians but soon es- caped. In 1814 he enlisted again. These were rough times, and not all of die fighting was with the enemy. For example, Lt. Chasteen Scott (1785-1861), under whom he served as a Sergeant, "whipped a rascally Yankee con- tractor [sic]" on 30 Oct 1814, the day that Gra- ham and the rest of the company were finally discharged from active service.3 Graham probably intended to return to his former occupation as a silversmith in which he had been occupied before the war. As he passed dirough Lexington, Kentucky, on his way back to Harrodsburg, he chanced to meet Dr. Benjamin Winslow Dudley (1785-1870) in Lexington. Dr. Dudley had received his M.D. at the University of Pennsylvania in 1806, and he had just returned from medical studies in England and France in 1814. He was to be particularly known for his success in operating for kidney and bladder stones; he is said to have operated on 225 such cases, all without benefit of anesthesia, losing only three pa- tients. He also performed successful cataract surgery. He opposed die use of bloodletting and was one of the first to sterilize his surgical instruments in boiling water. He is considered the founder of the medical school at Transyl- vania University in Lexington. Dudley, slightly younger than Graham, immediately offered him a chance to become one of his first stu- dents. Graham, just past die age of 30, was virtually uneducated, except in die ways of the woods, and war, and he told Dudley he lacked both the education and money to study at the university. Dudley replied he would keep the offer open till whenever it was wanted. After the war was over in the spring of 1815, Graham made another trip to New Or- leans. There he accepted an offer from James Hull, D.D., rector of Christ Church, to teach school, which he did, just barely keeping ahead of the students. By this means he likely gained die remedial skills he needed for his future college career. An outbreak of yellow fever, so common in New Orleans at that time, forced him to leave the city. He took a ship to New York, but the ship had its own outbreak, and a number of the people on it died; Gra- ham was ill when he arrived. The ship was quarantined, but he hired a skiff, landed in Virginia, and walked back to Kentucky by way of the Cumberland Gap. He then accepted die offer to study with Dr. Dudley, a turning point in his life. In 1810 Transylvania University was one of six medical colleges in die nation; by 1820 that number had grown to 26. When Craham be- 140 Christopher ( lolumbus ( Jraham— Dm all 111 -"&. '^g^t^^L^r ^i^t/fl r IsOt^-*^*-*^ c5f~a^ v -* is* Figure 1. Christopher Columbus Graham. From W. B. Alien. A History of Kentucky L872 142 Journal of 1 1 it- Kentucky Academy of Science 65(2) came a student of Dr. Dudley in 1817 it was still one of the best in the country. Graham's quick mind and unusual abilities were put to good use. This was the course of study, ac- cording to Wright: During this period requirements for the degree of doctor of medicine required a student to take two years of lecture courses, unless he had been a prac- ticing physician for four years, in which case he need- ed to attend only one year. All candidates had to be twenty-one years or older, write a thesis of not less dian twelve or more than forty pages on a designated medical subject, and pass two examinations, one be- fore the faculty and one before the president and trustees. The curriculum covered the areas of anat- omy and surgery, theory and practice of medicine, materia medico and medical botany, obstetrics, chemistry, and the institutes of medicine.'1 It has been remarked with some justifica- tion (though not entirely correctly) that Gra- ham did very little work in the profession in which he was trained.5 With an associate, Dr. Henry Miller, he practiced medicine in Har- rodsburg for five years following his gradua- tion and did at least some medical work for years afterwards. He wrote a medical book in 1866 titled The True Science of Medicine, ev- ery copy of which seems to have disappeared.6 He performed but one operation for kidney stones, and that successfully, on a small child.7 For over 30 years he was proprietor of Har- rodsburg Springs, the most famous "watering- place" in Kentucky at that time.8 Louis Jacob Frazee, M.D. (1819-1905), writing on the medical uses of the mineral waters of the state, said: Thirty years ago the Harrodsburg Springs under the management of Dr. Graham, was one of the most popular watering places in Kentucky. The beautiful grounds, and the fine hotel accommodations pre- senting the most attractive features of the place. The most important ingredient of die water here, is sul- phate of magnesia, which renders it aperient, but aside from this it possesses no very decided medicinal virtues. It may be used in toipor of the bowels, es- pecially when accompanied with dyspeptic symp- toms, indeed in almost every case where a gende ape- rient effect is desired. This has long since been aban- doned as a watering place.9 No doubt Graham did at least some con- sultation during that time, which was probably a factor in the popularity of die springs as a place of healing. He had on his staff in die IS 10s Dr. E. B, Thomas, a hydrotherapist.10 The hydrotherapy department had been or- ganized by Dr. Roland S. Houghton of New York, a famous teacher ol hydrotherapy" VanArsdall, after a lengthy analysis of the treatment at Harrodsburg Springs, concluded, "It is obvious that Dr. Graham must have practiced a mild sort of psychotherapy in as- sociation with the baths and general hygienic measures." vl In his old age Graham is listed in the census records as a "Retired Physician," which gives credence to die idea that he con- sidered medicine his primary occupation. Psychology, then called mental philosophy, was an area in which Graham studied that may be considered an offshoot of his medical in- terests. His The true science of mind (1869), is a devastating critique of the current "mental philosophy," and it is also his own attempt at a positive contribution to the field. He wrote: My natural turn of mind led me, in early life, to moralize upon all events, and caused a pleasure (no self-creation, take notice) in me to do just as I pleased, which was to read everything I could find upon the subject of mind. With this foundation I commenced my practical study of mind, and having for more than fifty years mixed with all nations and languages of the human family, from the native In- dian up, or rather, down, to die snobs, parvenus, and paragons — the fashionable folly of our race — have sought to find where happiness dwells.13 Unlike practitioners of the discipline in our day, Graham tended to draw an overt moral from the material he discussed. He related the following incident (reminding us, perhaps, of Pavlov's dog) about his pet, Fidel: I here relate a little incident that illustrates two great leading principles: I had a sprightly and inter- esting puppy, to which the cook often threw egg- shells, thus teaching it to eat eggs; the result being the breaking up of all my setting hens and loss of chickens. All this, however, my fond attachment in- duced me to put up with; but another branch of ed- ucation caused its death. Breachy sheep occasionally entering my yard, I set little Fidel after them, and was often greatly amused to see die little creature in full chase, tight and tight after a flock of great sheep; when on its return it would frisk around me, and looking up widi innocent laughter seemed to say: "I did what you told me; wasn't it funny?" By and by, however, till I thus lost ten or twelve by its example in learning others to help it, had I courage to take its life, knowing tiiat the fault was not in the dog, but in myself; nor had I myself a heart, or will, to perpetrate ( Ihristopher C i'n i M 1 1 ii , ( Jraham — Dm all 113 the deed, but hired another to do it. This illustrates two vital principles: L". The force of education for good or evil, even upon the brute; 2'1. The necessity of punishing, or taking the life, to prevent disastrous consequences, and to save the lives ol many. Parents .mil friends often think it smart to hear their little ones swear or commit innocent depredations, as I did little Fidel, not thinking it might lead to their de- struction." His criticism of "faculty psychology" was particularly acute. At one time as many as 40 "faculties" were admitted by some authors, and each author had a different list. Graham did not accept that series of errors. In a chap- ter titled "What is a Faculty?" he began by saying: Faculty is a word coined by the manufacturers of shoddy text-books on psychology, mental philosophy, or metaphysics — all meaning the same thing. . . . I5 The word, he said a little later, conveys a false meaning and is enforced by arbitrary author- ity.16 This is similar to current opinion on the subject. T H. Leahey, in the Encyclopedia of Psychology, wrote, "Strictly speaking, faculty psychology died in the 20th century, at least as regards scientific psychology. Under behav- iorism, psychologists became skeptical of in- ferred entities of any sort, including the mind, which therefore had no need of subdivi- sions." lv Graham said in the preface to his work, "I shall strive to drive innate ideas, as witches have been done, from the world, and to show that this thing called divine conscience is a parasite — an effect — is not a principle — has no separate existence from the prejudice and education of the mind. . . ." 18 His object, as he said, is the application of natural law to man, and this is certainly a scientific objective. He offered a one-line summary of his theory: "Cod has endowed us with sensibility, from which arise pleasure and pain, and conse- quently a desire or will to do or not to do! thus is resolved in a short sentence, the mighty question, the great enigma of psychology, soul, mind or intellect, all meaning the same thing." l9 The reason Graham spent so little time and effort in the field of medicine was probably because the medical profession at the time was badly paid, with the exception of some of the large practices of fashionable cits' doctors. But also he was extremely skeptical about the benefits thai medical practitioners could con- fer on their patients. The medical profession at the time was in disarray, and theories abounded. Before L850 there were published in this country alone 213 different medical journals, some of them surviving only a lew- issues.2" Stricllv speaking, there was no science of medicine, and anyone who was interested primarily in science could only look on the dis- cipline (or lack thereof) in despair. This situ- ation, in some fashion or other, extended to many of the allied sciences, such as chemistry. Graham remarked: The chemist, though aeting upon the necessary laws ot science, is as often disappointed in his results from the endless and unseen counteracting influences, as the man well acquainted with human nature is of the anticipations of his results. The physician, in like manner, is constantly perplexed and disappointed in the sequences of his prescriptions; for though calo- mel be a purgative, and tarter will puke, calomel may vomit, and tarter purge, from some necessary exist- ing, yet unseen, condition of system.21 A little further in The true science of mind he made an analogy between the medic and the "superficial metaphysician": The physician, when ignorant of those occult work- ings upon his patient, and pressed hard for explana- tion, treats the ease with deep gravity and most learned technicality; such as morbid irritation, normal and abnormal condition of system; loss of sensorial power, accumulated excitibility, revulsion, translation, concatenation, and above all, "vis medieatrix natura" is dragged in as the universal panacea of medical ig- norance.22 He quoted at length a well-known passage from the French physician Francois Magendie (1783-1855), called by one of his own stu- dents "the great sceptic." Graham quoted at some length from one of Magendie's famous lectures, which begins "Gentlemen: Medicine is a great humbug."-3 And this seems to mirror what Graham thought of the medicine of his day, that is, he realized it was not a science. Claude Bernard (1813-1878), the student of Magendie mentioned above, was the first to set medicine on the road to becoming an em- pirical science. Bernard wrote in his most im- portant book, published in Paris in 1865, "We are doubtless far from the time when all med- icine will be scientific; but that need not pre- vent our conceiving it possible. . . ." This may 144 journal nl the Kciituckv \cadeinv ol Science 65(2) scnf to justify Graham's doubts concerning the scientific nature ol medicine at the time.-' Graham's contributions to the progress of medicine, at least on the local level, wen- practical, and it may have been his practical nature that led to his more intrepid exploits in connection with the medical school. The post- mortem examination of an Irishmen, killed ln- one of his fellow countrymen in a quarrel, led to difficulties between Dr. Dudlev, the mentor of Graham, and Dr. Daniel Drake (1785- 1852), die professor of medical botanv and materia medica. Feelings on die controversy ran high enough that Dudlev challenged Drake to a duel, something fashionable at the time. Drake declined die honour, but his friend. Dr. Richardson, professor of obstetrics, accepted in his place. This was dulv arranged with Graham as die second of Dr. Dudlev; Graham even cut the gold buttons off the coat Dudlev had bought in France, so as not to proxide a target. At the first exchange of fire, Richardson, who missed, received a potential- ly fatal wound, which was immediately stanched by Dudlev, sa\ing his life. From diis time the two professors became fast friends.25 But what of the Irishman who caused all the trouble in die first place? There was no sense in allowing him to go to waste, and this led to what the newspapers of die time called "The Battle of die Graveyard."26 During the period Graham was at Transyl- vania, the French physicians were die most medically advanced in die world. Pathological medicine was their great contribution to ^'est- em medical science: the concept of tracing a disease or illness to a specific organ or part of the bodv. The introduction of pathology, diough dien in its infancy, was (for good or ill), the single greatest factor changing the course of medical research in the United States. Padiological medicine logically led to surgery as die means of finding and removing; die cause of illness. Dissection, which is nec- essary to train a good surgeon, required a sup- ply of cadavers, and, in die absence of any le- gal means of obtaining diem, was a major problem for medical science. Graham wrote years later drat he had "'headed all die resur- recting expeditions."27 He said, "I was Dud- ley's favourit [sic] and well I might be, and was his only dependence in procuring subjects, and was his demonstrator, often dissecting all night while others were out on pleasure."2 In 1S22. very near the time of "The Battle of the Graveyard," the following advertise- ment appeared in Wooler's British Gazette: Many hundred dead bodies will be dragged from dieir wooden coffins this winter, for the anatomical lectures (which have just commenced), the articula- tors, and for those who deal in the dead. . . . The violation of the sanctity of the grave is said to be needful, for the instruction of the medical pupil, but let each one about to inter a mother, husband, child, or friend, say shall I devote this object of my affection to such a purpose; if not. the onlv safe coffin is Bridg- man's Patent wrought-iron one. Da\id Burrell, in an interesting paper on die origins of undertaking, from which the preceding quotation was taken, remarks that Cincinnati, with sfx medical schools in die area, more than any other city in the West, was very concerned about body-snatching. He pointed out that the metallic coffin, intended to keep the body safe, may have actually aided the body-snatchers by keeping their quarry fresh longer. At any rate these intended final resting places were guarded bv walls and watchmen (the fence around the gravevard was never intended to keep the occupants in), and even with landmines, torpedoes, and spring guns that exploded when disturbed. As Burrell, who has written several papers on as- pects of this topic, savs: "Communities literally feared for their dead each time the medical schools began a new session."29 Graham said diat once his party was pur- sued near Nicholasville while making their way from a cemetery to dieir horses, and one of several shots lodged in the subject he was earning on his back. He wrote concerning die cause of the trouble between his professors. We were taking up the Irishman that caused the duel as above named, and again taking [taken] prisoners bv an armed guard and we hauled up to the court for trial but diere was no law to make the dead pri- vate property, so the declaration of the Scriptures, that from dust we came and unto dust we must re- turn let us off bv paving one cent damages for taking that much clay or soil.30 "The Battle of the Graveyard" led to what must have been an interesting court case. The defense was bv a top-notch lawyer, John Jor- dan Crittenden (1786-1863), who was gover- nor of Kentucky' from 1848 to 1850, just re- Christopher Columbus Gn mam Dinall I 15 signed from (lie U.S. Senate in March L819 to return to his private practice in Frankfort, One writer remarked. In all likelihood the judicial decision not to order the body's return did not hinge upon any "convincing" quote of Genesis . . . but because the right to sue lor something must inhere in a preceding riulit to control or own.31 This is probably a misunderstanding of the situation. The lawyer and the judge were ob- viously aware of Blaekstone's Commentaries (Book 4, Chapter 17) "Of Offences Against Private Property," which stated that among the Romans die stealing of a corpse (though in- decent) was no felonv unless one stole some Of the grave clothes with it. Genesis was read- ily admissible into am courtroom in Kentucky at this time, and die passage ". . . dust thou art, and unto dust thou shalt return" (Gen. 3: 19) was not irrelevant: it provided the means of settling the case while technically letting the plaintiffs win: Dust was cheap.32 Graham's scientific interests extended far beyond the field of medicine. Among the pa- pers of Henry Clay (1777-1852) is a letter from Dr. Graham written to Clay in 1825 when Clay was Secretary of State under Pres- ident John Quincy Adams (1767-1848). Gra- ham, who had already traveled fairly exten- sively in the West and Mexico, told Clay he was interested in any kind of government mis- sion diat would pay him expenses for travel. He hoped in this manner to serve his country as well as further his scientific interests: I am fond of the sciences of Natural History and mineralogy, which traveling so much extends. My cabinet is already respectable, and I wish to enlarge it.M In 1805 the entire mineralogical collection at Yale University fit in a single candle box. It, like the somewhat larger collection at Har- vard, was but recently acquired. In 1810 a Eu- ropean collection of about 10,000 specimens was on loan for public exhibit at Yale. This was finally purchased by the college in 1825, the year in which Graham already had a "respect- able" collection. A public subscription for the Yale collection was made, and the purchase price was $20,000. Scientific collections of ma- terials all over the country were scanty; for ex- ample. Harvard's herbarium was insignificant until the 1840s.34 The work of private collec- tors was important since the biological and earth sciences had nol vH completed collect- ing, describing, and classifying materials In the letter referred to above Graham re- minded Cla) they had met several times and had become acquainted a( Transylvania Uni- versity. (Clay was a trustee of thai institution. Graham also mentioned thai lie had become proprietor of Harrodsburg Springs, inviting ( !lay to visit next time he was in the area. He discussed some political matters relating to Andrew Jackson, always of interest to Clay. Then, in the last paragraph of the letter, he made the following statement: I should be pleased to know the probability of Mr. Rafinesque's success in his Banking schemes, as lie has flooded me with letters, appointing me sole ageni in all his operations. I know him to be so visionary, that I have given the subject but little attention.* There is a Iikelv connection between Gra- ham's interest in natural history and his ac- quaintance with Constantine S. Rafinesque (1783-1840). It may have been Grahams practical business instincts diat led Rafinesque to fix upon him as suitable to advance his scheme of divisible stock coupons. Graham's consideration of Rafinesque is in keeping with the general opinion of him at the time. Raf- inesque related that he visited Graham in the course of his travels in Kentucky: I visited again the small cabinet of Mai. Thompson at Shawnee Spring near Harrodsburg. that of Shells of Dr. Graham at Harrodsburg, and that of curiosities by Mrs. McDowell at Danville. I went on horseback with Dr. Graham to survey ancient monuments on Salt R. where we dug fossil teeth.37 Dr. Graham was a life-long naturalist. We have some of his descriptions of geological for- mations, one of which is the "Devil's Pulpit' along the Kentucky River in Nelson Countv Frederick Hall, M.D. (1780-1843), who visit- ed Graham's Springs in June 1837, wrote. I cannot help saving, that our polite landlord i^ ad mirably qualified for the management of this im- mense establishment. Doctor Graham is a well-in- formed physician, gentlemanly in his deportment, and exceedingly accommodating to all his boarders. His many kind attentions have brought me under lasting obligations to him. We have taken several long walks together. To-day he made me acquainted with an interestini: localih ol the sulphate of barvtes. It is about two miles south from his house. The substance 14fi Joiinml of (lie Kentucky Acadeim ol S cienc( 65(2) forms a vein, in secondary limestone rock, which is nearly perpendicular to the horizon, varying in thick- ness from one to eight inches, and extending, ac- cording to Dr. G. at least, twenty miles in length. The mineral is white, and its structure laminated.1'1 Graham explored caves and was an invet- erate collector of fossils and other natural ar- tifacts; in 1872 the large cabinet he collected was estimated to be worth $250,000. His ma- terial was donated to the Museum of the Ken- tucky Free Public Library (Louisville) at that time, and he began to assemble another col- lection. In his archaeological work at Big Bone Lick, Graham was not content merely to col- lect bones, but he formed a theory concerning die geology of the region, which I will not cov- er here; he described the valley and explained why it was sinking. Graham did not merely collect, like Prof. Rafinesque, but he also wrote accounts in which he attempted to set forth the significance of what he had found. Dr. Graham had been a close observer of wildlife from an early age. He once compared himself to John James Audubon (1785-1851), the famous naturalist and painter of birds, with no advantage to the latter: Audubon was never a closer observer of birds than I have been of all animals; so much so, indeed, that I have learned the language of many of them. I can tell by the voice of birds when they see a serpent as well as if I were to see it myself."10 In the early part of the 19th century the biological and earth sciences were still in die process of collecting, describing, and classify- ing materials. Rafinesque, often exuberant and careless in his methods, was among the first scientists of note in the West to engage in this necessary task on such a large scale. His meth- ods and manner left him open to criticism, so diat he did not always get credit for what he actually accomplished, but part of his legacy in the West — Kentucky at diat time — was his students. He is said to have been the first (if not the first, one of the early few) to actually use specimens in the classroom. He was an indefatigable traveler, collector, and writer. He attempted to found a botanical garden in Lex- ington, but the project came to naught. A pro- fessor of Rafinesque s caliber would have left a profound mark on a student such as Gra- ham. Even if Graham never sat in his formal lectures (though it is likely that he did), Tran- sylvania University was too small for such an influence to be unfelt. Graham, with his in- terest in nature and science, whatever reser- vations he might have had about Rafinesque's "visionary" tendencies, would have gravitated to a teacher with such a background and knowledge." Graham's work in geology and archaeology may have been his most visible contribution to scientific work in Kentucky. In 1871 the Public Library of Kentucky was incorporated by the Legislature. Former Gov. Thomas Bramlette (1817-1875), Graham's son-in-law, and Reu- ben T. Durrett (1824-1913), of the Kentucky Historical Society, who also married one of Graham's daughters, were the chief organiz- ers. They raised over $400,000 (by means of a lottery, though the word "lottery" was carefully avoided) to endow it. The next year a building for the library was purchased for $210,000. There were placed in it 40,000 volumes, and a Museum collection of 250,000 "specimens." Graham was named curator of die museum and was the chief donor of the artifacts. It was in this capacity that he conducted the exca- vations at Big Bone Lick. The Library itself soon went bankrupt, amid charges of fraud and mismanagement.4- Its effects went to die Polytechnic Society of Kentucky, and later to the Louisville Free Public Library. It may not be possible to even trace die materials at this point, since the Louisville Free Public Library later gave away all the artifacts to museums across the country, apparently without docu- menting where anything went.43 As late as 1884 Graham still had materials to sell and donate, as he wrote to John P. Knox (1844— 1903), the state geologist.44 Elsewhere I have published Grahams ac- count of his excavation at Big Bone Lick, Boone County, Kentucky; only a brief sum- mary is given here.45 For 30 days he dug with 10 men from the area who were paid one dol- lar a day. Graham was then 93 years old. He wrote not only about his own excavation but also provided a glimpse of prior digs in the area. He was fortunate to have as one of his workmen Thomas Rich (1808-1883), who had dug there on various excavations for years. Na- thaniel S. Shaler (1841-1906), mentioned here, was professor of geology at Harvard Uni- versity. He excavated at Big Bone Lick in 1863. Graham said in the article: Christopher Columbus Graham — Dm/ill i r In the year 1833 Thomas Rich, as above named, who dug for me. was foreman in excavating lor- an agent of New York, and exhumed a gigantic skeleton, twenty-two feet long and eleven feet high, with disks twelve feet long, all ol which are now in the Kentucky Department of the British Museum, in London. Mr. Rich told me that he also disinterred, during the same digging, the skeleton of an elk whose horns were seven feet long, since which he has been dig- ging for various parties, among whom was Mr. Shaler. our present geologist of Kentucky, but never found an entire skeleton ol any kind; and I to save money to those who may wish to search, say that I am sat- isfied there will never be another found. I obtained petrified horns ol both deer and elk, but not very large. There are several important things about Gra- ham's investigation. The most significant seems to be the following discovery, which he owed to Mr. Rich and the local workers: In my excavations we often came to piles of what 1 drought were collections of yellow, soft sand; but see- ing die old diggers rubbing it between their hands till warm, and smelling it with a grin and leer of the eye diat said to me you are green, I found it to be decayed flesh that still had the odor, as dogs around the pit would smell and scratch into it. I am not aware that any other excavator mentions die preserved remains of the flesh oi these creatures. If this is true, and I see no reason to doubt that it is, it would certainly be relevant in any inquiry concerning the age of the beasts whose bones have created so much interest. If conditions have continued to pre- serve this flesh in a state similar to the present, it might even be possible for archaeologists to acquire DNA samples from it, which would doubtless extend our knowledge on the whole subject of the animals inhabiting the Lick in prehistoric times. One of Graham's other discoveries of more local historical interest at Big Bone Lick is his finding the remains of the salt-boiling works, which ceased about 1812. He found them six feet below the surface, and he offered an ex- planation as to why the ground should be lit- tered with preserved bones (that is, hard and undecayed) when the Lick was discovered, but the bone-bed should be so far below the sur- face, 10 to 12 feet, in 1876. So far as finding artifacts is concerned, the excavation could probably be called a success, as Graham re- ported that he "brought off seven barrels of bniics. a number ol buffalo heads, and both mammoth and mastodon molars but found no very large bones or tusks 'bill had nine feet of a 14-foot tusk given me b) Mr, McLaughlin proprietor [of the hotel]), and left upon the ground a earl load of bones of various ani- mals."46 Bid it was significant in other ways, and it speaks well lor Graham as a scientist that he was concerned (o publish an account ol his findings rather than nierch earn awav bones, as was the habit of so many of the oth- ers who sought bones at the Lick. Dr. Graham was in communication with a number of eminent scientists. He wrote his account of the excavations at Big Bone Lick at the request of Prof. Fredric W. Putnam (1839-1915) of Harvard University. Professor Putnam became curator of the Peabodv Mu- seum of American Archaeology there in 1875. a year prior to the work at Big Bone; he held the position until 1909. Appointed professor of archaeology and ethnology' at Harvard in ] SS6, he is considered one of the first anthro- pologists to work in the United States. In 1874 he served as the assistant to the Kentucky Geological Survey under Dr. Shaler, and it was likely that he met Dr. Graham at this time. Graham also corresponded with Charles Darwin (1809-1882). In a four-page letter dat- ed 30 Jan 1877, he informed Darwin that he had defended him, John Tyndall (1820-1893), and others against the attacks of a clergyman.47 It seems Graham also sent him a copy of the article on Big Bone Lick, which had been pub- lished in the Courier-Journal (Louisville) on 29 Jan 1877, and this was probably the real reason for the letter. Darwin is said to have commented favourablv on the article and to have passed it on to Thomas Henry Huxlev (1825-1895) and others, who made similar comments on it. There are two short letters Graham wrote in 1880 (28 March and 17 April) in which he informed Darwin that his earlier letter had been framed and was to be placed in the Kentucky statehouse. No doubt this was to be in the fireproof rooms that hail been given the Kentucky Historical Society, ot which Graham was a charter member. In 1S20 Graham married Theressa Sutton, daughter of the proprietor oi the Harrodsburg Springs, and soon owned the resort himself. Here is how Frederick Hall, himself a medical doctor, described it: 148 Journal <>l the Kentuck) Acaderm ol Science 65(2) Our stage drove directly to the hotel nlv one to follow their father into tlit- medical profession. In ISo2 Craham was illegally arrested by Union soldiers, one of whom he severely wounded, as a Confederate sympathizer. He was imprisoned for a week and then released, lie published The true sci- ence of medicine in 1866 and The true philos- ophy of mind in L869.54 The next year began his association with the Public Library of Ken- tucky. Richard II. Collins (182^1889) pub- lished the two-volume History of Kentucky in 1874 and acknowledged the contributions of Dr. Graham to his research. From this point until his death in 1885 Graham became more and more involved in the history of Kentucky, writing letters to people such as Lyman C. Diaper (1815-1891), one of the foremost col- lectors of manuscripts and other materials for the history of the West.55 In addition Graham wrote a series of articles about the early pio- neers of Kentucky, which appeared in die Louisville Monthly Magazine (1879). In 1880 Graham became a charter member of die Kentucky Historical Society; indeed, it is probably significant that his name is the first on die list in the charter granted by the state legislature. When he was 90 he still went on frequent expeditions for geological specimens, which would have included the excavation at Big Bone Lick. Col. E. C. Walton, the 50-year editor of the Stanford Interior Journal, re- membered that Graham, at age 97, walked the 10 miles from Crab Orchard to Stanford to eat a birthday breakfast with a friend.56 For his 100th birthday his friends in Louisville held a centennial celebration at the Louisville Hotel, consisting of a huge pioneer dinner and many, many toasts, all of which was written up and occupied most of the front page of the Cou- rier-Journal along with a wood-cut showing Graham as a venerable-looking gentleman with long white flowing hair. This event was felt by many of those present, and his acquain- tance, to be the passing of an age, since he was the last living link with the early past of Kentucky, and he had been acquainted with most of the pioneers of note.57 Upon the grounds of the Harrodsburg Springs — the sole remaining relic of the era when the resort flourished under Dr. Gra- ham— is a tombstone marked "Unknown." It is not the stone of Craham. At Ins death on 3 Feb L885 he was buried in the cemeter) in Danville, nol far from the place of the pioneer fort in which he was born on 10 Ocl 1784. Il is perhaps fitting that the stone of one who began his scientific career robbing graves should have been stolen. ' The stone on the grounds at the Springs marks the grave oi a society girl of the south who \ isited and. after dancing all night in the fashionable ballroom, fell (lead in the morning. ' This romantic in- cident seized the public imagination, and passed into Kentucky legend, forever to be as- sociated with Graham's Springs. It max serve to remind us that there are many things we would like to know about Craham and the people of this era, but also there is much we owe them.61 The past, though vanished, still lives in the collective memory through the writing and reading of history and offers us insight into the present. ENDNOTES 1. Thomas D. Clark. Tlic Kentucky (1942: rpt. Lexing- ton: University Press of Kentucky, 1992), p. 223. A $10,000 was reward offered and advertised in Europe and America to anyone who could out-shoot Graham. 2. G. Glenn Clift, Notes on Kentucky veterans of the War of 1812. (Anchorage, Kentucky: Borderland Books, 1964). p. 342. 3. Kentucky Soldiers of The War of 1812. Report of the Adjutant General of the State of Kentucky. (Frank- fort: E. Polk Johnson. 1891). He is reported to have served from 1 Jan 1814 till 31 Oct 1814. There is some indication diat this was his second period of sendee in that war. 4. John D. Wright, Transylvania, Tutor To the West (Lexington: Transylvania University, 1975; qit. Uni- versity Press of Kentucky, 1980), p. 84. 5. Russel Blaine Nye, Society and Culture in America. 1830-1860. (New York: Harper & Row. 1974 1. p. .342 remarks that in this period nearly all physicians were general practioners. and the income was not high. As late as 1873 there were hut 149 hospitals in the entire country, and a country doctor might make $3000 a year. 6. Extensive searches have been made for this work. 1 would like to thank my colleagues Jinny Ussel and Michelle Foster, of the Boone Count) Public Library, Union, Kentucky, for doing an exhaustive internet search lor this hook, ami Maggie lleran ol the Lloyd Library, Cincinnati, lor searching their collection ot medical tests for the same vllsheler unite ol this book in 1933: "En>l KCT suckers occurs else- where, too, as around a healthy staminate tree in a home yard in Middlctown, Ohio; if die suckers there had not been removed, the yard would have been a "coffeetree thicket" (David M. Brandenburg pers. coiiiin. 2003). Suekering is not confined to staminate trees: a living pis- tillate tree in Boone County, Kentucky, produces them (Richard Feist pers. comm. 2004 1. KCT has been known as a wild-growing plant and as a cultivated tree for at least 250 years. Why, then, is the suekering habit so often not noted in literature and by- some individuals who work with trees? Are only certain KCT genotypes prone to suekering? Under what condi- tions does KCT suekering occur? Is it a trait expressed only after a concatenation of certain environmental con- ditions? Does die removal of suckers within a day or two of their appearance contribute somehow to their contin- ued production? These are questions yet to be answered. In eastern North America other trees, too, produce suckers. Well known among these are tree-of-heaven (Ai- lanthus altissima), papaw (Asimina triloba), hackberry (Celtis occidentalism trembling aspen (Populus tremulo- ides), black locust (Robinia pseudoacacia), and sassafras (Sassafras albidum). Leaves. In common with juvenile leaves of various other trees, diose of KCT suckers differ from mature leaves (Figure 1). On sucker shoots and seedlings of KCT the first one to nine leaves are usually 1-pinnate, 6 to 45 em long, and with 3-12 pairs of pinnae. Later growth pro- duces an increasing number of pinnately compound pin- nae, starting widi often one in a median position on the rachis of an otherwise 1-pinnate leaf. On some leaves a simple pinna or two may be produced between compound pinnae. At the tip of primary and secondary rachises of die leaves of KCT may be an inconspicuous, soft brisde. Those I have seen do not exceed 1 cm, but Halsted (10), calling them "tendrils," reported diat diose on die primary rachis may reach 2.5 cm. What die bristies represent is uncertain, although they may simply be extensions of the rachis or "degenerate terminal leaflets" (5) much as can be seen in black walnut (Juglans nigra; Juglandaceae). Similar brisdes may occur also on leaves of honey-locust (Gleditsia triacanthos; Fabaceae), a tree related to KCT. Though frequendy described as lacking, die stipules of KCT, when present, are represented by caducous, minute scales or bristles. Stipels are similar to stipules and, like them, are not always present. Specimens vouchering diis note are deposited in the herbarium of Northern Kentucky University (KNK). I thank David M. Brandenburg, The Dawes Arbore- tum, Newark, OH, and Richard Feist, Burlington, KY, for data. 154 Notes loo Figure 1. Silhouette of selected leaves from a Kentucky coffeetree sucker with nine leaves, showing change in leaf morphology from the base to the tip of the sucker. Omitted are leaves 3, 5, and 7. each of which was closely similar to the leaf immediately below it. (a) Leaf 1, one-pinnate, (b) Leaf 2, one-pinnate, (c) Leaf 4. one-pinnate, (d) Leaf 6, mostly one-pinnate but with a median pinna compound, (e) Leaf 8, with four median pinnae compound. (0 Leaf 9. mostly two-pinnate, with all pinnae but die basal two pairs compound, the leaf with essentially the same morphology as mature leaves of the species. The vertical bar = 10 cm. LITERATURE CITED. (1) Bailey, L. H. 1944. The standard cyclopedia of horticulture. The Macmillan Com- pany, New York, NY. (2) Bates, E. J. S„ C. R. Blinn, and A. A. Aim. 2002. Regenerating quaking aspen, http:// www.extension.umn.edu/distribution/naturalresources/ DD5637.html. Accessed 28 Aug 2004. (3) Dirr. M. A. 1998. Manual of woody landscape plants. 5th ed. Stipes Publishing, Champaign, IL. (4) Anonymous. 1902. Cof- feetree (Gymnocladus dioicus). U.S.D.A. Forest Serv. Circ. 91. (5) Elwes, II. J., and A. H. Henry. 1906. The trees of Great Britain and Ireland. Vol. 2. Privately pub- lished. Edinburgh. (6) Wilbur, R. L. 1963. The legumi- nous plants of North Carolina. North Carolina Agric. Exp. Sta. Techn. Bull. 151. (7) Robertson, K. R., and Y.-T. Lee. 1976. The genera of Caesalpinioideae (Leguminosae in the southeastern United States. J. Arnold. Arbor. 57:1-53. (8) Steyermark, J. A. 1963. Flora ol Missouri. Iowa State Univ. Press, Ames, 1A. (9) Environment Canada. 2003. Species at risk, http://www.spetiesatrisk.gc.ca/search speciesDetails-e.cfm?SpeciesID = 222. Accessed in Vug 2004.(10) Halsted, B. D. 1902. The "tendrils" of the K, n tucky coffee-tree. Torreya 2:5-6. — George F. Buddcll II, 8790 Napoleon Zion Station Road, Dry Ridge. k\ 41035. L56 [ournaJ ol the Kentuck) Vcadem) o) Science 6 Vegetative Proliferation in Eragrostis minor (Little Lovegrass; Poaceae). — Unusual or malformed plant parts arc occasional!) seen in man) groups ol plants. These distortions, termed tcratological, max occur in veg- etative structures, such as leaves or stems, or in repro- ductive structures, that is. Rowers and fruits ( 1. 2). Darwin (3) discussed morphological deviations in his Origin oj Species, calling them "monstrosities." which he thought would be injurious or at least not useful, stating that such could grade into "varieties" if the condition was inherited by subsequent generations. The replacement of various parts in the spikelets of grasses by vegetative growth has been known at least since it was reported in 1620 bv Bauhin (4) in the species now- known as Poa bulbosa L., and in 1690 bv Rav (5) in what is now known as Festuca vivipara (L.) Sm. Many other species of grasses have been reported to exhibit similar conditions. In a s\nopsis paper on die subject, Beede (6) reported such replacement of parts in about SO different grass species, many of which have been given varietal or form epithets such as "\i\ipara." "prolifera." or even "monstrosa." A fairlv widelv distributed example is Poa bulbosa L. (bulbous bluegrass), known by most people only from its proliferative form. P. bulbosa ssp. cicipara (Koel.) Arcangeli, though a non-proliferative form is sometimes seen (7). Beede (6) recognized diree hpes of vegetative struc- tures in grass inflorescences: proliferation, in which die paleas and lemmas are replaced by leaf-like structures; phyllody, in which the paleas and lemmas are replaced widi such well-developed leaves that thev are differenti- ated into sheadis and blades; and true civipary, in which a seed germinates precociously on the parent plant to give rise to a new individual (8). The terms viviparv and pro- liferation have been used almost interchangeably and without discrimination in the literature for several hun- dred vears, but Arber (9) argued that the term "\i\iparv" should be restricted to conditions where seeds germinate and grow in situ. The causes of vegetative proliferations in grasses are variable, and may include one or several of the following (6, 10, 11, 12, 13): heritable causes, such as hybridization and pokyploidv: malformations (teratology), caused bv me- chanical injury, chemical damage, or attacks bv patiiogens or pests; and adverse environmental conditions, such as in water levels, light levels, high altitude or latitudes, and abrupt changes in day length or temperature. Vegetative proliferations have been reported for several species of die genus Eragrostis (love grasses). Beede (6) listed three species of the genus in which die condition is known: E. brizoides (L.f.) Nees, E. capensis (Thunb.) Trin.. and / mrescens PresI; in addition, proliferations reported by Jain 14 in E. angetica (Roxb. Steudel. I have recentl) seen several specimens of Eragrostis mi- rtoi from Kentucky, Michigan, and Ohio: see Specimens Examined with abnormal infloresci nces and since I was unable to uncover am literature record ol such a condi- tion in this grass species further investigation was war- ranted. Eragrostis minor Host Little lovegrass; Poaceae; Fig. I V IB), also known as Eragrostis poaeoides Beaux, ex Roemer 6c J. A. Schultes, is a widely distributed, weedy species introduced from Europe. It is found in nearly all of the lower 4S states in the United States ( http://plants. usda.gov/), often along railroad tracks (15, 16), or road- sides and paths (17). It is similar to E. cilianensis (All.) Janchen (stink grass; = E. megastachya (Koel.) Link from which it differs by ha\ing lemmas 1.5-2 mm long 2-2. S in E. cilianensis). ha\ing andiers 0.2 mm long (0.5 in E. cilianensis). and often lacking glands on the keel of the lemma (usually present in E. cilianensis > (18, 19). The proliferations in spikelets of E. minor (Fig. 1C) fit best the definition of "phvllodv" as given bv Beetle (6i. In some spikelets, all parts, including glumes, have become leaf-like, die smaller appearing as blades, and the larger differentiated into sheaths and blades. Some spikelets have only a single floret replaced bv a proliferation, while other spikelets are completely replaced bv what appears to be a plandet. In diese proliferative structures, no sexual parts are present even diough remaining florets appear normal and contain andiers and/or pistils. It is unknown whether the vegetative proliferations in E. minor are able to function as propagules. Manv plant species along the railroad tracks at the Ken- tucky and Ohio sites (see Specimens Examined) showed evidence of herbicide treatment. It is possible that this is die cause of die unusual morphology of some spikelets from these populations, though this remains to be tested. Specimens examined. KENTUCKY: Mason County; Marysville; weedy in railroad vard. 3 Jul 2001. M. A. Yin- cent 9562. J. W. Thieret, it W. M. Vincent (KNK, MU). MICHIGAN": St. Joseph Counrv; Three Rivers; weedv ground at railroad crossing, 17 Aug 1995, A. W. Cusick 32692 (MICH, MU). OHIO: Butler County; Oxford; weedy along railroad line, 19 Jul 2004, M. A. Vincent et al. 12159 (DAY. DOY. ISC. KNK. MO. MICH. MU. OSH. US, UTC). I am grateful for assistance from the following people in tiiis study: Thomas Lammers (OSH), Shane Shaw (MU), and John W. Thieret (KNK). LITERATURE CITED. (1) Goebel, K. 1900. Organ- ography of plants. Clarendon Press. Oxford, England. (2) Figure 1. A. Whole plant of Eragrostis minor, showing inflorescence widi mixed normal and proliferative florets (Vincent 12159, MU; scale bar = 2 cm). B. Inflorescence showing normal florets (Vincent 12159, MU; scale bar = 1 cm). C. Portion of an inflorescence showing proliferative florets (Cusick 32692, MU; scale bar = 1 cm). Notes L57 15S al of the Kentucky Acacli I Science 65(2) Guedes, M.. and 1'. Dnpuv. 1979. Teratological r i ic >< I i f it ;i - tions and the meaning ul (lower parts. Today & Tomor- row's Printers & Publishers, New Delhi, India. (3) Oar- win, C. 1872. The Origin of species. 6th ed. Carlton House, New York, NY. (4) Bauhin. C. 1620. Prodromos theatri botanici. P. Iacobi, impensis I. Treudelii, Franco- furti ad Moenum. (5) Ray, J. 1690. Synopsis methodica stirpium britannicarum. Samuel Smith, London. England. (6) Beetle. A. A. 1980. Vivipary, proliferation, and phvl- lody in grasses. J. Range Managem. 33:256-261. (7) Cus- ick, A. W., and M. A. Vincent. 2002. Poa bulbosa ssp. bulbosa (Poaceae) in North America. Michigan Bot. 41: 19-22. (8) Elmqvist, T, and P. A. Cox. 1996. The evolu- tion of vivipary in flowering plants. Oikos 77:3-9. (9) Ar- ber, A. 1934. The Gramineae. A study of cereal, bamboo, and grass. Macmillan, New York, NY. (10) Filatenko, A. A. 1969. Teratological changes of the flowers resulting from the remote hybridization in die genus Triticum L. Bot. Zhurn. 54:153-155. [In Russian.] (11) Harmer, R. 1984. Vegetative proliferation and vivipary in Scottish grasses. Trans. Bot. Soc. Edinburgh 44:261-268. (12) Lee, J. A., and R. Harmer. 1980. Vivipary, a reproductive strat- egy in response to environmental stress? Oikos 35:254— 265 (13) Nielsen, E. 1941. Crass studies. V Observations on proliferation. Bot. Gaz. 103:177-181. (14) Jain, S, K. 1968. Notes on Indian grasses — X: Proliferation in Era- grostis P. Beauv. and Bromus L. Bull. Bot. Surv. India 10: 229-230. (15) Brandes. O. 1993. Eisenbahnanlagen als Untersuchungsgegenstand der Geobotanik. Tuexenia 13: 415-444. (16) Peterson, P. M. 2003. Eragrvstis Wolf. Pag- es 65-105, volume 25, Flora of North America Editorial Committee (eds). Flora of North America North of Mex- ico. Oxford Univ. Press, New York, NY. (17) Northam, F. E.. R. R. Old, and R. H. Callahan. 1993. Little lovegrass (Eragroatis minor) distribution in Idaho and Washington. Weed Technol. 7:771-775. (18) Gleason, H. A., and A. Cronquist. 1991 . Manual of vascular plants of Northeast- ern United States and adjacent Canada. 2nd ed. New York- Botanical Garden, Bronx, NY. (19) Hitchcock, A. S., and A. Chase. 1950. Manual of die grasses of the United States. 2nd ed. USDA Misc. Publ. 200. United States Gov- ernment Printing Office, Washington, DC. — Michael A. Vincent, W. S. Turrell Herbarium (MU), Department of Botany, Miami University, Oxford, Ohio 45056. J. Ky. Acad. Sci. 6S(2):159-162. 2001. INDEX TO VOLUME 65 ABBOTT, J. RICHARD, 94 ABSTRACTS. 59-63 Acer, 119 A. negundo, 135 A. rubrum, 22 A. saccharinum, 135 A. saccharum, 134 Achyranthes japonica, 97 Vcipenser fulvescens, 77. S3. 91 Aconitum unoinatum, 97 Aegopodium podagraria, 97 Agave, 48 Agkistrodon contortris mokasen, 24 Agricultural Sciences, 59-60 Akebia quinata, 1 10 Alismataceae, 47 Alismatidae, 47 Allegheny woodrat, distribution of, 33-38 habitat use of, 33-38 in a mixed-mesophytic forest. 33- 38 Alopecurus pratensis, 97 Amaranthaceae, 97 Ambystoma jeffersonianum, 30 American beech. 22 American crow, 21, 23 American robin, 31 American toad, 29 Amphibians. 27, 29 Amur honeysuckle. 27-32 ANTONIOUS. GEORGE K, 59, fiO Anura, 29 Apiaceae, 97 Araceae, 47 Archaeological excavations. 12-20 at Owl Cave, 12-20 Mammoth Cave National Park, 12-20 Arecaceae, 47 Arecidae, 47 Aristolochiaceae, 98 ARNOLD, S. BROOK. 33 Aroids, 47 Arsenic, in tap water, 62 from private wells. 62 in central Appalachia, 62 Ash, white, 134 Asimina triloba. 22 Asteraceae, 59. 97 Atriplex liortensis, 46 A. littoralis. 46 AWARDS 2003. 5.5-58 Azadiraclitin. movement in soil, 60 BAKER, MICHAEL D.. 21 RASKIN. CAROL, 53 BASKIN, JERRY, 53 Rasswood. 134 Baylisascaris procyonis, 33 BE BE. FRED N!. 61 Beech, 134 Berea College Forest, vascular flora in, 107-131 BERTRAM. LONNIE R„ 33 Beta-actin genes, 104-107 Bittemut hickory, 134 Black holes, gravitational tensing of. 62 Black oak, 22 Black rat snake, 29 BLALOCK, RICHARD. 62 Blight, chestnut, 33 Blue jay, 23 Bluegrass, bulbous, 156 Bluestem, little. 137 Bhmtnose darter, 80 BOATENG, DADDY N„ 59 Boehmeriopsis pallida, 67 Bog lemming, southern, 34 BOOK REVIEW, 54 Boone Count)- Kentucky, 132 natural terrestrial vegetation of, 132-139 Botrvchium, 46 Bouteloua curtipundula, 137 Box turtle, eastern, 29 Box turtles, 29 Boxelder, 135 Brassicaceae, 97 BREWER, KEVIN S., 33 Broad-headed skink, 30 Bromeliaceae, 47 Bromeliads, 47 Brown snake, 29 Brown-headed cowbirds, 21, 23 BRYANT, WILLIAM S.. 132 BUDDELL. GEORGE E, 51 BUDDELL, GEORGE E. 155 Bufo americanus, 29 Bufonidae, 29 Bulbous bluegrass, 156 Bullfrog, 29 Burbot, 80 Cacalia suavoeolens, 97 Cactaceae, 46 Calcium, in plasma ol rats. 61-62 Cambarus cumberlandensis, 78 C. laevis, 108 Camelina microcarpa, 97 Campanulaceae, los CAMPBELL, NATALIE J.. 33 Caprifoliaceae, 27-32 Capsicum annuum Bell Boy, 59 Cardamine impariens 97 Carex, 17 C. pedunculata, 97 < \RSTE\S KENNETH C. 12 Carya, 119 C. cordiformis, 134 C ovata, 134 C. spp., 22 < iaryophyllaceae, loo Caryophyllidae 16 Castanea pumila var. pumila, 104 Catonotus, SI Cat-tails, 47 Caudata, 29 Cavefish, spring so Celastraceae, 1 10 Celosia argentea, 46 Centra] newt. 29 Centrarchidae, 104-107 Chenopodiaceae, 46 Chenopodiaceae, 97 Chenopodium ambrosioides, 46 C. botrys, 46 C. pumilio, 97 C. spp., 46 Chestnut blight. 33 Chestnut oak. 22. 1 19 Christopher Columbus Graham. 149-153 Chrvsosplenium americanum, 98 CICERELLO, RONALD R., 76 CLARK. AMY M., 33 Cla\tonia caroliniana, 46 CLEMONS, CHAD M.. 33 CLEMONS, JOE E.. JR.. 33 Clinostomus elongatus, 78 C. funduloides, 7s COE, STEVE, 62 Colubridae, 29 Colubrids. 29 Commelinaceae. 47 Commelinidae, 47 COMPTON, MICHAEL C, 76 < (AIPTON, R. N., 5 COMPTON, STEPHEN, 62 COOPER, ANN-SIMONE, los COOPER, NIGEL C. F 57 COOPER, ROBIN L. los Copper, in plasma of rats. 61-62 Copperhead. 24 Comtis florida, 22 Corvus brachyrhynchos, 21. 23 Cottonwood, 135 Cowbell Reservoir, 1 17 < Irangon crangon, 1 11 Crataegus coccinea, Mil Crayfish, 7s ins Crow, American. 21 Cryphonectria parasitica I I Cucurbita, 12. 17 Cyanocitta cristata, 23 ( yperaceae, 17. 97. loo 159 lfi() Journal oi the Kentucky Acaderm of Science 65(2) Cyperus, 47 Cspraceae, 98 Cyprinella glactura, 78, 88 Cypripedium calceolus var. parviflo- rum, 98 C. parviflorum, 98 Dace, mountain redbelly, 79 redside, 78 rosyside, 78 Darevskia valentini, 1 Darter, bluntnose, 80 duskytail, 82 fringed, 81 johnny, 90, 91 Deer, white-tailed, 15-16 Dendroica discolor, 24 D. virens, 24 DERTING, TERRY, 62 Desmognathus fuscus, 29-30 DICK. D. KYLE, 33 Dioscorea quatemata, 48 D. villosa, 48 d-iron complex, Disporum maculatum, 98 Distinguished college/university sci- entist award, 57-58 DURTSCHE, RICHARD D., 27 Dusky salamander, 29 Duskvtail darter, 82 DUVALL, JAMES, 140 Dysphania, 46 D. anthelmitica, 46 Eastern box turtle, 29 Eastern garter snake, 29 Eastern redcedar, 137 Ecoregions map, Kentucky's Level IV, 60 EISENHOUR, DAVID J., 76 Elaphe obsolete, 29 Eleocharis, 47 Empidonax virescens, 24 Emydidae, 29 Epigean cravfishes. growth of, 108- 115 Equisetophyta, 116, 120 Eragrostis brizoides, 156 E. capensis, 156 E. cilianensis, 156 E. megastachva, 156 E. minor, vegetative proliferation in, 156-158 E. poaeoides, 156 E. virescens, 156 Eriophorum virginicum, 98 Esox masquinongy, 77 Etheostoma chlorosoma, 80 E. crossopterum, 90, 91 E. nigrum, 81 E. olivaceum, 81 E. percnurum, 82 E. squamiceps, 81 Eumeces fasciatus, 3 E. laticeps, 3, 30 Euonymus alata, L10 Eurasian milfoi, 27 Eurecea cirrigera, 29-30 European wall lizard. 1—4 Fabaceae, 104, 154 Fagaceae, 46, 104 Fagus, 119 F. grandifolia, 134 Fast LDA calculation, modeling ni- tride crystals, 85-93 Fatoua aspera, 67 F. japonica, 67 F. \iIlosa, 67-75 Faxonius propinquus, 110 FERNANDO, SHAMANTHI, 62 FERNER, JOHN YV., 1 Festuca \ivipara, 156 Fishes, Kentucky, distributional re- cords of, 76-84 FLEMING, CHRIS A., 116 Floerkea proserpinacoides, 98 Flora of North America, comments on, 39^0 Flowering dogwood, 22 Food choices, 61 Forbesichthys agassizii, 80, 82 Forest songbirds, nesting success of, 21-26 in mixed mesophvtic forests, 21- 26 in Kentucky, 21—26 Fraxinus, 119 F. americana, 134 FRAZIER, DONALD T.. 56 Fringed darter, 81 Frog, green, 29 Frogs, true, 29 Frogs and toads, 29 GALBRAITH, SHANNON L., 132 Garter snake, eastern, 29 GELIS, RUDY A., 94 Gentiana fiavida, 98 Gentianaceae, 98 Geography, 60 Geraniaceae, 98 Geranium dissectum, 98 Geum laciniatum, 104 Grama, side-oats, 137 Grass, Indian, 137 stink, 156 Gravitational lensing, of black holes, 62 Green frog, 29 Guidelines for Contributors to the Journal, 64—65 Gymnocladus dioicus, 154-155 Gypsy modis, 33 Hamamelidae, 46 HAWKINS, LISA, 60 Health Sciences, 60-62 HELD, MICHAEL E., 132 Heracleum lanatum, 104 II maximum, Mil Herbicide movement, in soil water, 59 Herpetofauna biodiversity, 27-32 I lexastylis heterophylla, 98 II. virginica, 98 Hickories. 22-23 Hickory, bittemut, 134 shagbark, 134 IIORRALL, AUDREY D., 33 Hybognathus ha\i, 79 Hydrocharitaceae, 100 Hylocichlia mustelina. 31 Iclithyomyzon ammoecetes. 77 I. bdellium, 77 I. fossor, 77 I. unicuspis, 77, 81 Indian grass, 137 Italian wall lizard, 1^1 Jackson County, vascular flora in, 107-131 Jefferson salamander, 30 Johnny darter, 81 JOHNSON, DIANE, 55 JONES, NATALIE J.. 33 JORDAN, CLARENCE, 59 Juglans, 17 Juncaceae, 47 Juncus, 47 Juniperus \irginiana, 137 Kalmia laufolia, 23 Kentucky coffee tree, 154—155 Kentucky fishes, distributional rec- ords of, 76-84 Kentucky- vascular plants, 94-103 Kerria japonica, 110 KOCHHAR, TEJINDER S., 59 KORNMAN, LEWIS E., 76 KRUPA, JAMES J., 33 Kudzu, 27 Lacertidae, 1—4 LACKI, MICHAEL J., 21 Lake sturgeon, 77 Lamprey, Ohio, 77 silver, 77 Lardizabalaceae, 110 LAUDERMILK, ELLIS L., 76 La\th\Tus hirsutus, 104 Lemming, southern bog, 34 Lemna valdiviana, 47 Lepomis cyanellus, 104-107 L. macrochirus, 104-107 Ligustrum obtusifolium, 110 Liliaceae, 48, 98 Liliopsida, 120 Limnanthaceae, 98 Linaceae, 108 Lindera benzoin, 22 Linum usitatissimum, 108 Liparis loeselii, 10S Index to Volume fi5 L61 Liriodendron, 1 19 L. tulipifera, 22, 134 I., tulipifera, 134 l-iron complex, Little bluestem, 137 Little lovegrass, 156-158 LLOYD, CHRISTOPHER M., 33 Lobelia nuttallii, 108 Lonicera niaackii, 27—32 Lota lota, 89, 91 Lovegrass, little, 156-158 Lower Silver Creek Reservoir, 117 Ludwigia hirtella, 96 Lungless salamanders, 29 Lvciuni barbarum, 102 Lycopersicon esculentum "Moun- tain Spring," 59 L. hirsutnm, 59 f. glabratum, 59, 60 f. hirsutum, 59, 60 L. pennellii, 59, 60 L. pimpinellilolium, 59, 60 Lycopodiaceae, 96 Lycopodiella appressa, 96 LyeopodiopliN'ta, 116, 120 Lycopodium appressum, 96 Lymantria dispar, 33 Lysimachia vulgaris, 10S Lvtlmnn salicaria, 27 Madison County, vascular flora in, 107-131 Madtom, slender, 79 Magnoliophyta, 116, 120 Magnoliopsida, 120 Magnolophvta. 46, 47 MAHURIN, S. M., 5 Maianthemum, 48 Malvaceae, 100 Manfreda, 48 Maple, silver, 135 sugar, 134 MARLETTE, MARTHA A., 61 MATHIS, CALEB, 62 McCLURE, S. B„ 5 McEVOY, NICHOLAS L„ 27 Meadow vole, 34 MEIJER, WILLEM, 52-53 Microtus pennsylvanicus, 34 Mite, two-spotted spider, 60 Mniotilta varia, 24 Mognoliidae, 46 Molothrus ater, 21 Monolepis nuttalliana, 46 Moraceae, 67 Mows alba, 102 Mountain redbellv dace, 79 Mountain-laurel, 23 Mulberry Weed, 67-75 MUNSIF, SIDDHARTH, 62 Muskellunge, 77 MUTISYA. SAMUEL M„ 60 \IYKA, JENNIFER LEICH, 63 Myosoton squaticum, 100 Mvrioplivlluin spicatum, 27 Neotoma magister, distribution of, 33-38 habitat use of, 3.3-38 in a mixed-mesnphvtic forest, 33 38 Nesting success, of forest songbirds, 21-26 Newt, central, 29 Newts, 29 NCYVANG, HERMINE, 61 Nitride crystals, fast LDA calcula- tion to model, 8.5-93 Nocomis effusus, 78 Northern red oak, 22, 134 NOTE, 51 NOTES, 154-158 Notopthalamus viridescens, 29-30 Notropis telescopus, 78 N. maculatus, 79 Notuws exilis, 79 Oak, chestnut, 22, 1 19 northern red, 134 white, 134 Oak-hickory forest, 22 Oenothera linifolia, 108 Ohio lamprev, 77 OKONNY, ESUGHANI. 61 Oleaceae, 110 Onagraceae, 96, 108 Ophioglossaceae, 46 Orchidaceae, 48, 98, 100, 108 Orconectes australis australis, 112 O. australis packardi, 108-115 O. cristavarius, 108-115 O. inermis inermis, 112 O. propinquus, 110 OTIENO, TOM, 63 Outstanding Academy service award, 56-57 Outstanding college/university sci- ence teacher award, 55 Outstanding secondary school sci- ence teacher award, 55—56 Owsley Fork Reservoir, 118 Palms, 47 PANEMANGALORE, MYNA. 61 Papaver dubium, 108 Papaveraceae, 46, 108 Parity-violating energy difference. 5-11 PATTERSON, MATTHEW A., 59, 60 Pawpaw, 22 PCR-based Fl Hybrid Screen, 104- 107 Pepper sweet, 59 Peromyscus leucopus, 62-63 Pest control, phytochemicals for, 59 PEYTON, DAVID K.. 104 Phoxinus oreas, 79, 82 Physics and Astronomy, 62 Physiology and Biochemistry, 62-63 Phytochemicals, for pest control. 59 Pinophyta, 1 16, 120 r s, U9 P. echinata, 23 P. virginiana, 23 Pinus-Liriodendron-Quercus com- plex. 110 Plant-based laboratory project. 63 Platanthera integrilabia, 10S Plethodon cinereus, 29-30 P. glutinosus, 27, 29-3] P. richmondi, 29-30 Plethodontidae, 29 Poa bulbosa, 100, 156 ssp. vivipara, 156 Poaceae. 47. 97 Podarcis muralis, 1—4 P. muralis muralis, 1 P. pityusensis vedrae, 2 P. sieula, 1^1 Polygonaceae, 104 Polygonum densiflowm, 103. 113 Polypodiophvta, 116, 120 Poplar, tulip! 134 Populus deltoides. 135 Potamogeton, 47 Potomogetonaceae, 47 PRATER, CALLIE A., 33 Primulaceae, 108 Procambarus clarldi, 113 Procyon lotor, 33 Pueraria montana, 27 Purple loosestrife, 27 Pyrethrins, on pepper fruits, 59 on tomato fruits, 59 Quercus, 119 Q. alba, 22, 134 Q. coccinea. 22 Q. prinus, 22 Q. wbra, 22. 134 Q. velutina, 22 Rabbits, 15 Raccoon, 33 Raccoon roundworm, 33 Raccoons, 15 Rana catesbeiana, 29-30 R. clamitans, 27, 29, 31 Ranidae, 29 Ranunculaceae, 46, 96. 97. LOO Ranunculus ficaria, 100 R. parviflows, 96 R. pusillus, 96 R. sceleratus, 96 Rat snake, black. 29 RAUBENHE1MER. DIANNE, 63 Ravine salamander. 29 Red Lick Reservoir, lis Red maple, 22 Red oak. northern, 134 Redbacked salamander, 29 Redbellv dace, mountain, 79 162 il of the Kentucky Academy oi Science 65(2) Redcedar, eastern, 137 Redside dace, 78 Reptilia, I A, 29 Rhamnaceae, 1(1(1 Rhamnus firangula, Km Rhododendron, 23 Rhododendron maximum, 23 Rhynochospora, 47 RIVERS, RORERT C, 61 Robin, American, 31 Rosaceae, 104, 110 Rosyside dace, 78 Roundworm, raccoon, 33 Rubiaceae, 100 Salamander, dusky, 29 Jefferson, 30 ravine, 29 redbacked, 29 slimy, 29 southern two-lined, 29 Salamaders, 29 lungless, 29 Salamandridae, 29 Salicaceae, 51 Salicornia depressa, 46 S. europes, 46 Salix exigua, 51 Salsola collina, 46 Sandbar willow, 51 Saxifragaceae, 98 Scaphirhynchus platorynchus, 78, 91 Scarlet oak, 22, 119 Sceloporus undulatus, 3 Schizachyrium scoparium, 137 Schoenoplectus, 47 S. tabemaemontani, 47 School lunch, nutritional benefit, 61-62 School lunch plate waste, 61 Science Education, 63 Science instruction, enhancing in middle schools, 63 Scirpus, 47 S. fluviarilis, 100 S. validus, 47 Scrophulariaeeae, 96 Seiurus aurocapillus, 24 Serpentes, 29 Setophaga ruticilla, 24 Sewage sludge, in land farming, 60 Shagbark hickory, 134 SHAW, CHARLES P., 33 Sherardia arvensis, 100 SHIRER, JOHN G., 62 Shiner, taillight, 79 whitetail, 78 Shortleaf pine, 23 Shovelnose sturgeon, 78 Shrimp, 114 Sida hermaphrodita, 100 Side-oats grama, 137 Silcnr ovata, 100 Silver lampre) 77 Silver maple, 135 Skink, broad-headed, 30 Slender madtom, 79 Slimy salamander, 29 Smilacina, 48 Snake, black rat, 29 brown, 29 eastern garter, 29 Snakes, 27, 29 SNYDER, JOHN C, 60 Soil water, herbicide movement in, 59 Solanacedae, 102 Solonaeeae, 59 Sorgastrum nutans, 137 SOTO, MARGARET M„ 63 Southern bog lemming, 34 Southern two-lined salamander, 29 Spicebush, 22 Spider mite, two-spotted, 60 Spinacia oleracea, 46 Spiranthes lucida, 100 Spring cavefish, 80 Squamata, 29 Squash, 17 Squirrels, 15 Stellaria aquatica, 100 Stink grass, 156 Storaria dekai, 29 Stuekenia, 47 S. filiformis, 47 S. peetinatus, 47 Sturgeon, lake, 77 shovelnose, 78 Stygobitic crayfishes, growth of, 108-115 Sugar maple, 134 Sunfishes, 104-107 SURMONT, ALBERT, JR., 76 Sweet pepper, 59 Synaptomys cooperi, 34 Taillight shiner, 79 Tanacetum einerarilolium, 59 TEMPLETON, SUSAN B., 60, 61 Terrapene Carolina, 27, 29-30 Terrestrial vegetation, of Boone County, Kentucky, 132-139 Testudines, 29 Tetranychus urticae, 60 Thamnophis sirtalis, 29-30 THIERET, JOHN W„ 51 THOMPSON, MATTHEW, 63 THOMPSON, RALPH L., 94, 116 Thrush, wood, 31 Tilia americana, 134 Toads, 29 Tomato, 59 Tradescantia zebrina, 17 True frogs, 29 Tulip poplar, 134 Turdus migratorius, 31 Turtle, eastern box, 29 Turtles. 27. 29 Turtles. 29 box. 29 water, 29 Two-spotted spider mite, 60 Typhaceae. 47 Upper Silver (.'reek Reservoir, 117 Urtiea japonica, 67 VALENTINE, ANGELA M., 33 Vallisneria americana, 100 Vascular flora, in Jackson County, 107-131 in Madison County, 107-131 in the Berea College Forest, 107- 131 Vascular plants of Kentucky. 94-103 Veronica polita, 96 VINCENT, MICHAEL A., 67 Vireo olivaceus, 24 V. solitarius, 24 Virginia pine, 23 VIRK, MANINDER, 62 Vole, meadow, 34 Walnut, charred, 17 Wandering jew, 47 Water turtles, 29 White ash, 134 White oak, 22, 119, 134 WHITE, BRENT C, 55 White-footed mice, health of, 62 anthropogenic disturbance on, 62-63 Whitetail shiner, 78 WHITSON, MAGGIE, 54 Willow, sandbar, 51 Wilsonia citrina, 24 Wood thrush, 31 Woodrat, Allegheny, distribution of, 33-38 liabitat use of, 33-38 in a mixed-mesophytjc forest, 33- 38 WORKMAN, JEFFREY W, 33 YACEK, HENRY F JR., 21 Yellow-poplar, 22 Y'ODER, G, 94 Zebrina pendula, 47 Zinc, in plasma of rats, 61-62 Zingiberidae, 47 Zoology laboratory activities, in fresh- man biology majors course, 63 ZOURARAKIS, DEMETRIO P.. 60 Guidelines for Contributors lo the Journal 1. GENERAL Mil he con- A. Original research/review papers in science wi sidered for publication in JKAS; at least the first author must be a member of the Academy. Announcements, news, and notes will be included as received 13. Acceptance ol papers for publication in [KAS depends on merit as evaluated by each ol two or more review- ers. C. Papers (in triplicate) may be submitted at any time to the editor. John W. Thieret Department of Biological Sciences Northern Kentucky University Highland Heights. KY 41099 Phone: (859) 572-6390; Fax: (859) 635-3490 E-mail: thieretj@exchange.nku.edu List in the cover letter your telephone/FAX numbers, your E-mail address, and the names, addresses, and telephone numbers of two persons who are potential reviewers. D. Format/style of papers must conform to these guide- lines and also to practices in recent issues of JKAS. which are, in effect, a style manual. E. Papers should be submitted in hard copy. Do not sta- ple pages together. F. Indent the first line of each paragraph (but not the first line of entries in the Literature Cited). 2. FORMAT A. Papers should be in 12-point type on white paper 8.5 X 11 inches, with margins at least 1 inch all around. Double-space throughout the paper (i.e., one full line of space between each two lines of text, literature cit- ed, or tabular data). Do not justify right margins. B. Except for scientific names of genera and of infrage- neric taxa. which should be tvped in italics, die same type (roman) should be used throughout (i.e., one type size only; bold only for paper title). C. Sequence of sections in papers should, where appro- priate, be as follows: title of paper, name/address of author(s), abstract, bodv of paper, footnotes, table cap- tions, figure captions (all the preceding on consecu- tively numbered pages), tables, and figures. D. The running head (top right) should give nanie(s) of audior(s), a short version of paper title, and page num- ber of total. E. The first page should include the running head and. centered near the top of the sheet, the paper's title and the name and address of author(s). These should be followed immediately by the abstract. (The first page should look as much as possible like the first page of articles in JKAS.) F. The abstract, not to exceed 200 words, should be con- cise, descriptive, and complete in itsell without refer- ence to the paper. (;. The body ol the paper should, where appropriate in elude the following sections: Introduction Materials and Methods, Results, Discussion, Summary, Acknowl- edgments, and Literature Cited. I I. No mure than three levels of headings should be used: level 1. in capitals, centered; level -. in capitals/low- ercase, flush left; level ■'!. in italics, a paragraph indenl with initial capital onlv i except proper nouns and ad- jectives), and followed b) a period, the text then start- ing alter one blank space. I Personal communications (avoid il possible should be indicated in the text as follows: (name, affiliation, pers. eomni.. elate), e.g.. (O.T. Mark. Wainw right College pers. comm.. 5 Jim 1995). 3. STYLE A. In text, spell out one-digit numbers unless they arc- used with units of measure (four oranges, 4 cm) and use numerals for larger numbers; do not begin am sentence with a numeral. B. Use no footnotes except those for title page and tables. Footnotes, identified by consecutive superscript num- bers, should be entered on a separate sheet. C. Measurements should be in metric and Celsius units Define lesser-known symbols and give the meaning of acronyms at first use. Express time of das in the 24- hour system. Dates should be written day, month (ab- breviated to three letters), year without internal punc- tuation. Units with multiple components should ha\e individual components separated bv a virgule (e.g., g/ m2 or g/m2/yr). D. Names of authors of binomials may be included but onlv at the first mention of die binomial. Cultivar names are not italicized but are enclosed in single quotes. E. Useful guides for contributors to JKAS are die follow- ing: Scientific style and format: the CBE manual for authors editors, and publishers, 6th ed., Cambridge University Press, 1994: The Chicago manual of style, 14th ed.. University of Chicago Press, 1993; The ACS style guide, American Chemical Society, Washington, DC, 1986; and All' sli/le manual. American Institute of Physics, New York, 1990. 4. IN-TEXT CITATION OF LITERATURE A. Cite publications in the text b\ author(s) and date — e.g., (Readley 1994); multiple citations should l>c in alphabetical order and separated by semi-colons — e.g., (Ashley 1987; Brown 1994; Foster 1975': multiple ci- tations of works by one author(s) should be in chro- nological order — e.g.. (|oucs 197S. 1KIS31: publications by one author(s) in the same vear should be distin- guished In a. b, c. etc.— e.g.. (Smith 1994a. 1994b . For in-text references to works with one or two authors use names of both authors — e.g., [ones and Williams 1991); for works with three or more authors use name 163 US4 il the (Centuck) A.cadem\ <>l Science 65 J of the first author followed In et al. — e.g., (Lee el tl 19S5). B. Do not include an) reference unless it lias been pub- lished or accepted for publication ("in press" sei I" low). 5. LITERATURE CITED A. List all authors of each entry. Do not abbreviate jour- nal titles; abbreviations for these will be supplied by the editor. B. The first line of each reference should be txped (lush left; the remaining lines should be indented. C. Examples of common types of references are given below. JOURNAL ARTICLE Lacki. M.J. 1994. Metal concentrations in guano from a gray bat summer roost. Transactions of the Kentucky Academy of Science 55:124-126. BOOK Ware, M., and R.VV. Tare. 1991. Plains life and love. Pi- oneer Press, Crete, WY. PART OF A BOOK Kohn. J.R. 1993. Pinaceae. Pages 32-50 in J.F. Nadel (ed). Flora of the Black Mountains. University of Northwestern South Dakota Press, Utopia, SD. WORK IN PRESS Groves, S.J., I.V. Woodland, and G.H. Tobosa. n.d. De- serts of Trans-Pecos Texas. 2nd ed. Ocotillo Press, Yucca City, TX. 6. ILLUSTRATIONS FIGURES (LINE DRAWINGS, MAPS, GRAPHS, PHO- TOGRAPHS) Figures must be camera-ready, glossy, black-and-white prints of high quality or laser prints of presentation qual- ity. These should be designed to use available space ef- fectively: a full page or part of one, or a full column or part of one. They should be mounted on heavy white board and covered with a protective sheet of paper; pho- tographs to be grouped as a plate should have no space between them. Dimensions of plates must observe page proportions of the journal. Each illustration in a plate may be numbered as a separate figure or the entire plate may be treated as one figure. Include scale bars where appro- priate, l ottering should be large enough i" be legible after reduction; us,' lowercase letters for sections ol a figure. Figure captions should be self-explanator) without refer- ence to the texl and should be entered '>u a page separate from the text. Number figures in Arabic numerals. Statis- tics presented in figures should be explained in die caption (e.g., means are presented ± SE, n = 71. TABLES Each table and its caption must be double-spaced, num- bered in Arabic numerals, and set on a sheet separate from the text. The caption should begin with a title relat- ing the table to the paper of which it is a part; it should be informative of the table's contents. Statistics presented in the table should be explained in the caption (e.g., means are presented ± SE, n = 7). Table should be sub- mitted in hard copy only; they need not be included on a disk. 7. ETHICAL TREATMENT OF ANIMALS AS RE- SEARCH SUBJECTS If vertebrate or invertebrate animals are involved in a re- search project, the author(s) should follow those guide- lines for ethical treatment of animals appropriate for the subjects, e.g., for mammals or for amphibians and reptiles. Papers submitted to JKAS will be rejected if their content \iolates either die letter or the spirit of the guidelines. 8. PROOFS Authors are responsible for correcting proofs. Alterations on proofs are expensive; costs will be assessed to authors. Proofs must be returned to the editor within 3 days after the author receives diem; delay in return may result in delay of publication. 9. REPRINTS Forms for ordering reprints will be sent to the autiior when die proofs are sent. They are to be returned direcdy to Allen Press, not to the editor. 10. PAGE CHARGES Pages charges are assessed to audiors of papers published in Journal of the Kentucky Academy of Science. 11. ABSTRACTS FOR ANNUAL MEETINGS Instructions on style of abstract preparation for papers presented at annual meetings may be obtained from the editor. Copies will be available also at each annual meet- ing of die Academy. NEWS Kentucky Heritage Land Conservation Fund The mission of the Kentucky Heritage Land Conservation Fund (KHLCF) is to award funding to purchase and preserve selected natural areas in the Commonwealth; to protect rare and endangered species and migratory birds; to save threatened areas of natural importance; and to provide natural areas for public use, outdoor recreation, and education. Established by the 1994 Kentucky Legislature, KHLCF is administered by a 12-member board appointed by the governor. The board can award grants to acquire and protect areas of natural significance to local governments, state colleges/univer- sities, and specified state agencies. Special consideration will be given to the funding of agencies working together to meet the listed goals. All acquisition applications, along with comprehensive management plans, must be submitted to and approved by the board. The year 2003 was another eventful year for the KHLCF board. During 2003 it received and reviewed a large number of applications and approved 14 projects in 13 counties. In 2003 local government projects were approved in Calloway, Clark. Green, Harrison, Livingston, Logan, and Oldham counties; state agency projects were approved in Franklin, Garrard, Hardin, Harlan, Larue, and Letcher counties. The board also approved the Pine Mountain Trail State Park and provided initial acquisition funds. The trail will traverse Bell, Harlan, Letcher, and Pike counties. Since the first awards were made in October 1995 the board has approved 116 projects in 56 Kentucky counties. Almost 20,000 acres have been purchased. The fund is supported by the state portion of the unmined minerals tax, environmental fines, the $10 additional fee to purchase a Kentucky nature license plate, and interest on the fund's assets. For more information, contact the Department of Natural Resources, Commissioner's Office, 663 Teton Trail, Frankfort, KY 40601. The phone number is (502) 564-2184; the e-mail address is www.heritageland.ky.gov. AMNH LIBRARY 00167231 Journal Ql of Science.. American Museum of Natural History CONTENTS Received on: 02-08-05 ARTICLES Spread of Fatoua villosa (Mulberry Weed; Moraceae) in North America. Michael A. Vincent 67 Distributional Records of Selected Kentucky Fishes. Michael C. Compton, David J. Eisenhour, Ronald R. Cicerello, Lewis E. Kornman, Albert Surmont Jr., and Ellis L. Laudermilk 76 Developing a Fast LDA Calculation to Model Group HI Nitride Crystals. G. Vbder 85 Noteworthy Vascular Plants From Kentucky: A State Record, Range Extensions and Various Species of Interest. J. Richard Abbott, Ralph L. Thompson, and Rudy A. Gelis 94 A PCR-based Fj Hybrid Screen using the Beta-actin Genes from the Sunfishes Lepomis cyanellus and L. macrochirus (Centrarchidae) David K. Peyton 104 Growth of Stygobitic (Orconectes australis packardt) and Epigean (Orconectes cristavarius) Crayfishes Maintained in Laboratory Conditions. Ann-Simone Cooper and Robin L. Cooper 108 Vascular Flora of Five Reservoirs in the Berea College Forest, Madison and Jackson Counties, Kentucky. Ralph L. Thompson and Chris A. Fleming 116 Natural Terrestrial Vegetation of Boone County, Kentucky: Classification, Ordination, and Description. William S. Bryant, Shannon L. Galbraith, and Michael E. Held 132 Christopher Columbus Graham: Kentucky Man of Science 140 NOTES Notes on Root Suckers and Leaves of Kentucky Coffeetree (Gymnocladus dioicus; Fabaceae) in Kentucky. George F. Buddell II 154 Vegetative Proliferation in Eragrostis minor (Little Lovegrass; Poaceae). Michael A. Vincent 156 News Inside back cover List of Recent Reviewers 75 Index to Volume 65 158 CONTENTS AMNH LIBRARY 100 67231 Journal oj of Science.. American Museum of Natural History Received on: 02-08-05 ARTICLES Spread of Fatoua villosa (Mulberry Weed; Moraceae) in North America. Michael A. Vincent 67 Distributional Records of Selected Kentucky Fishes. Michael C. Compton, David J. Eisenhoui; Ronald R. Cicerello, Lewis E, Kornman, Albert Surmont Jr., and Ellis L. Laudermilk 76 Developing a Fast LDA Calculation to Model Group HI Nitride Crystals. G. Vbder 85 Noteworthy Vascular Plants From Kentucky: A State Record, Range Extensions and Various Species of Interest. J. Richard Abbott, Ralph L. Thompson, and Rudy A. Gelis 94 A PCR-based Fj Hybrid Screen using the Beta-actin Genes from the Sunfishes Lepomis cyanellus and L. macrochirus (Centrarchidae) David K. Peyton 104 Growth of Stygobitic (Orconectes australis packardf) and Epigean (Orconectes cristavarius) Crayfishes Maintained in Laboratory Conditions. Ann-Simone Cooper and Robin L. Cooper 108 Vascular Flora of Five Reservoirs in the Berea College Forest, Madison and Jackson Counties, Kentucky. Ralph L. Thompson and Chris A. Fleming 116 Natural Terrestrial Vegetation of Boone County, Kentucky: Classification, Ordination, and Description. William S. Bryant, Shannon L. Galbraith, and Michael E. Held 132 Christopher Columbus Graham: Kentucky Man of Science 140 NOTES Notes on Root Suckers and Leaves of Kentucky Coffeetree (Gymnocladus dioicus; Fabaceae) in Kentucky. George F. Buddell II 154 Vegetative Proliferation in Eragrostis minor (Little Lovegrass; Poaceae). Michael A. Vincent 156 News Inside back cover List of Recent Reviewers 75 Index to Volume 65 158 I H I I I v . ■ ■ I