DIAGNOSTIC OPAL PHYTOLITHS FROM PODS OF SELECTED VARIETIES OF COMMON BEANS: (PHASEOLUS VULGARIS) Steven R. Bozarth Numerous distinctly hooked silicified hairs are produced in pods of common beans (Phaseolus vulgaris). Similar phytoliths were found in six dicot species native to the central Great Plains in an extensive reference collection of bath cultivated and wild plants, Statistical analysis demonstrates that many of the hooked hairs produced in ‘common beans are significantly wider near the tip than thase produced in the other species. Bean phytoliths were identified at a late prehistoric village in central Kansas based on this distinguishing characteristic. ind comunidad prehistdrica en uw estado de Kansas central (E.E.U.U). Growing plants absorb water containing dissolved silica. Microscopic silica bodies are produced by the partial or complete silicification of plant cells, cell walls, and intercellular spaces. Silica bodies with characteristic shapes are called opal phytoliths, which form in most plants and are produced in many shapes and sizes. Phytoliths are diagnostic when their shapes and/or sizes are specific to a particular taxon. Many phytolith types are resistant to weathering and are preserved in soil over long periods of time. ‘One aspect of phytolith research is the identification of cultigens in archaeological sites. The ability to identify cultigens is important because macrofossils, the parts traditionally used to identify cultigens, are not preserved in most sites unless accidentally carbonized. The goal of this study is to determine if beans can be identified in prehistoric villages in the central Great Plains based on phytolith analysis. Four species of beans (Phaseolus) have been identified based on macrofossil analysis of archae- ological sites in the New World: P. vulgaris, the common, haricot, navy, French, or snap bean; P. coccineus, the runner or scarlet runner bean; P. /unatus, the lima (large seeded) and Sieva (smalll- seeded) bean; and P. acutifolius war. latifolius, the Tepary bean (Evans 1976; Kaplan 1965). The common bean, P. vulgaris, was the only Phaseolus prehistorically cultivated in the central Great Plains; the earliest documented evidence dates to A.D. 1070 (Adair 1988). Phytolith identification of beans, as well as maize, squash, pumpkins, gourds, and sunflowers in prehistoric villages largely depends on the production of diagnostic phytoliths in their seed or seed- bearing structures. These are the parts of the cultigens that would have been taken to the villages (Bozarth 1986). There are no reports of plants native to North America producing hooked silicified hairs. This includes analysis of grass phytoliths by Bonnett (1972), Brown (1984), Suess (1966), and Twiss et al. (1969); analysis of arboreal phytoliths by Brydon et al. (1963), Geis (1973), Klein and Geis (1978), Norgren (1973), Rovner (1975), and Wilding and Drees (1971, 1973, 1974) mnalysis of dicotyledonous herbs by Rovner (1975) and Bozarth (1985). However, ‘Steven R. Bozarth, Department of Geography. University of Kansas, Lawrence, KS 66045 American Antiquity, $5(1), 1990, pp. 98-104, Copyright © 1990 by the Society for American ArchaeologyREPORTS. 99 METHODS This study is based IS GSS > cultigen section consists of phytoliths extracted from the following: seeds and pods of shicld-figured beans and Kentucky Wonder pole beans (P. vulgaris); seeds, husks, and cobs of seven varieties of com (Zea mays); seeds and disks of three varieties of sunflowers (Ielianthus annus var. macrocarpa), seeds, rinds, and peduncles of five varieties of squash and pumpkins (Cucurbita pepoand C. maxima), seeds, rinds, and peduncles of two varieties of bottle gourds (Lagenaria siceraria); and the leaves of tobacco (Nicotiana rustica and N. quadravalis). All of these cultigens are nonhybrid, traditional varieties. The noncultivated reference section consists of phytoliths from the complete aerial portion and roots of 22 species of Poaceae; representative aerial portions (leaves with petioles and stem sections from various parts of the plants plus part of the inflorescence when available) of four species from other monocot families and 62 species of herbaceous dicots; leaves from 19 species of arboreal dicots; and twigs with needles from 10 species of gymnosperms. Most of these reference species were selected for study because they are among the more common species native to the central Great Plain: These include all species of Aristolachaceae, Moraceae, and Urticaceae and at least one of the more common species of each genus in Boraginaceae and Leguminoseae native to the central Great Plains. A representative aerial Portion and pods of P. polystachios also were processed. P. polystachios is the only Phaseolus species native to the Great Plains, where it occurs only infrequently; it has not been reported in Kansas (Barkley 1977). Phytoliths were extracted from the reference plants with a procedure modified from Rovner (1971, 1972), using Schulze’s solution (three parts 70 percent nitric acid and one part saturated potassium chlorate) to oxidize the reference material. ANALYSIS All of the above reference samples were studied at 400 with a research-grade Zeiss microscope. Numerous hooked silicified hairs were found in pods of the two varieties of common beans (P. vulgaris), the aerial portion of wild beans (P. polystachios), bog hemp (Boehmeria cylindrica), Minois tickclover (Desmodium illinoense), pellitory (Parietaria pensylvanica), and pipe vine (Aristolochia tomentosa). Hooked hairs infrequently occurred in leaves of red mulberry (Morus rubra) and only very rarely in wild bean pod: from each of these reference materials, except for wild bean pods, which produced too few phytoliths (only two hooked silicified hairs were produced in two complete wild bean he F test (a comparison of variances) demonstrates that, at the .01 level of significance, the width of the parent populations of hooked hairs produced in the two varieties of common beans are not the same as the populations of those produced in the other species. The Archaeological Test To test the feasibility of using phytoliths to identify beans in prehistoric villages, phytoliths were isolated from six sediment samples collected at site 14MN328, a Great Bend aspect village located0 AMERUCAM ANTIQUITY {Wel 88, No, 1, 1990) c d Figure 1. Silicified lnrs from (¢) pod of shield-figured bean (P. vulgaris); (8) pod of Kentucky Wonder pole ‘bean (P. ralgaris); and (c) aerial portion of bog hemp (Bochmeria cylindrica), (d) Fossil siicfied hair from bean (Phaseolus vulgaris) isolated from trash-filled storage pit at site 14MIN328, a Great Bend village in central Kansas. Photomicrographs taken at 100%, scale is 20 microns.REPORTS: 101 Table 1. Statistics o Microns from Tip) of 30 Hooked Silicified Hairs from Reference Species. ‘Sample Standard Range Deviation(s) Phaseolus vulgaris 210 2.06 Kentucky Wonder pole beans pods Phaseolus vulgaris 2-10 243 Shield-Agured beans ods Phaseolus polystachios 2a 263 n Wild bean aerial portion Aristolochia tomentosa 4 283 8 Pipe vine aerial portion Bochmeria evlindrica 4 20 45 Bog hemp aerial portion Desmodiwm ilinoense 26 307 14 Illinois tickelower aerial portion Morus rubra 36 40 64 Red mulberry leaves: Parietaria pensytvanica 4 223 1” Pellitory acrial portion in central Kansas. Five of these samples were from four trash-filled storage pits and one was from a hearth. These features were in an area that had never been cultivated. This site was selected because charred beans have been recovered from other Great Bend sites in central Kansas (Adair 1988), suggesting that beans likely were grown by the inhabitants of 14MN328. Phytoliths were isolated from the sediment samples using a modified version of the heavy-liquid flotation and centrifugation procedure reported by Rovner (1971). Phytoliths from each of these samples were mounted on microscope slides in immersion oil under 24 x 40 mm cover glasses and sealed with finger nail polish. Fach slide was scanned thoroughly at 400. Three hook-shaped silicified hairs were found in the isolates from a sample collected in the bottom (120 cm below surface) of a trash-filled storage pit. Two of these phytoliths were six microns wide (Figure Id) and one 4 microns wide measured six microns from the tip of the silicified hair. To determine if these fossil phytoliths were produced in beans, their widths were compared to the expected width distribution of reference species at one, two, and three standard deviations (Table 2). One standard deviation (1s) describes the expected width of 68.3 percent of the phytoliths. Similarly, two standard deviations (2s) describes the expected width of 95.4 percent of the phytoliths, and three standard deviations (35) describes the expected width of 99.7 percent of the phytoliths. AL 2s, the expected distribution of Illinois tickclover and red mulberry (the reference species with the closest expected distribution to common beans) are .8-5.4 microns and 2.7-5.3 microns, re- spectively. This demonstrates that there is less than a 2.3 percent chance that the two fossil phytoliths with widths of six microns were produced in Illinois tickclover or red mulberry. There is virtually no chance that they were produced in any of the other non-Phaseolus reference materials. Therefore, based on the reference materials studied, these phytoliths are almost certainly from beans. The fossil hooked phytolith of four microns cannot be identified as bean based on its width, since the expected102 AMERICAN ANTIQUITY [ol 5, No. 1, 1990] Table 2, Expected Distribution of Width (6 Microns from Tip) of 30 Hooked Silicified Hairs at 1, 2, and 3 Standard Deviations (6) from Reference Species. Is 2s 38 Phaseolus vulgaris 3475 139.5 116 Kentucky Wonder pole beans pods Phaseolus vulgaris 227. 09.5 o-11.9 Shield-figured beans pods Phaseolus polvstachios 1933 0 1241 S48 Wild bean aerial portion Aristolochia tomentosa 19-32 13-38 oad Pipe vine aerial portion Boehmeria cylindrica 1524 11-29 0-33 Bog hemp aerial portion Desmodium illinoense 19-42 85.4 0-65 Ilinois tickclover aerial portion Morus rubra 3447 275.3 21S. Red mulberry leaves Parietaria pensylvanica 15-30 73.8 O45 Pellitory acrial portion distributions at 1s for Illinois tickclover and red mulberry are 1.9-4.2 microns and 3.44.7 microns, respectively. However, it was most likely produced in a bean pod since 14MN328 is a fairly flat upland site, a habitat unsuitable for either Hlinois tickclover or red mulberry (Barkley 1986). ‘The identification of these three hooked hairs as Phaseolus is supported by the absence of this type of phytolith in a control sample collected in the site at a depth that represented the ground surface at the time of occupation. The phytolith assemblage from the control sample is representative of the native vegetation growing in the study area when the site was occupied. The absence of hooked hairs in the other archaeological feature samples is further evidence that this type of phytolith was not produced in the native vegetation at the site. SUMMARY AND COMMENTS statistical comparison of these phytoliths demonstrates that many ‘Beans were identified at 14MN328, a Great Bend village in ‘central Kansas based on statistical analysis of the width six microns from the tip of fossil hooked hairs isolated from the sediment sample collected near the bottom of a trash-filled storage pit. This identification is supported by the habitat, distribution, and relative rarity of other species that produced this type of phytolith in addition to the lack of hooked hairs in a control sample collected at the site. This study of hooked silicified hairs should be useful in the study of prehistoric agriculture as it provides a new method for identifying beans in archaeological sites in the central Great Plains, Phytolith analysis may be the only way to identify beans in most archaeological sites since beanREPORTS +103 macrofossils, the traditional part used to identify beans, are not preserved unless carbonized acci- dentally. Analysis of appropriate reference collections from other areas will determine if this phytolith classification can be used to identify beans in archaeological sites outside the region. Acknowledgments, Lam grateful to Ralph Brooks for giving me permission to sample plants at the University of Kansas Herbarium and for verifying the species identification of plants that were collected in the field. 1 would like to thank William Lees, Kansas State Historical Society, for partially funding this study. I also am ¢grateful to Dan Zwiener, University of Wisconsin-Madison, and the Living History Farms, Des Moines, Iowa, for providing samples of the common beans analyzed in this study. Gloria Rojas, University of Kansas, kindly translated the abstract into Spanish. REFERENCES CITED Adair, M. J 1988 Prehistoric Agriculture in the Central Plains. Publications in Anthropology 16. University of Kansas, Lawrence, Barkley, T. M. (editor) 1977 Atlas of the Flora of the Great Plains. The Great Plains Flora Association, lowa State University Press, ‘Ames. 1986. Flora of the Great Plains. The Great Plains Flora Association, University Press of Kansas, Lawrence. Bonnett, 0. T. 1972 Silicified Cells of Grass: A Major Source of Plant Opal in Illinois Soils. Agricultural Experiment Station 2. University of Illinois, Urbana. 1985 Distinctive Phytoliths from Various Dicot Species. Paper presented at the 2nd Phytolith Conference, University of Minnesota, Duluth. Brown, D. A 1984. Prospects and Limits of a Phytolith Key for Grasses in the Central United States. Journal of Archae- ological Science 11:345-368 Brydon, J. E., W. G. Dore, and J. S, Clark 1963" Silicified Plant Asterosclereids Preserved in Soil. Proceedings of the Soil Science Society of America 27:476-477. Madison, Wisconsin. Evans, A. M. 1976 Beans: Phaseolus spp. (Leguminosae-Papilionae). In Evolution of Crop Plants, edited by N. W. Sim- monds, pp. 168-172, Longman, London Geis, 1.W, 1973 Biogenic Silica in Selected Species of Deciduous Angiosperms. Soil Science 116: Kaplan, L. 1965" Archeology and Domestication in American Phaseolus (Beans). Economic Botany 19:358-368 Klein, R.L., and J. W. Geis 1978 "Biogenic Silica in the Pinaceae. Soil Science 126(3):145-155, Norgren, J 1973 ' Distribution, Form and Significance of Plant Opal in Oregon Soils. Unpublished Ph.D. dissertation, Department of Soil Science, Oregon State University, Corvallis. Piperno, D. R. 1988 | Phytolith Analysis: An Archaeological and Geological Perspective. Academic Press, San Diego. Rovner, 1 1971 Potential of Opal Phytoliths for Use in Paleoecological Reconstruction. Quaternary Research 1:343~ 359. 1972 Note on a Safer Procedure for Opal Phytolith Extraction. Quaternary Research 2:591 1975_ Plant Opal Phytolith Analysis in Midwestern Archaeology. Michigan Academician 8:129-137 Suess, E. 1966 " Opal Phytoliths. Unpublished Master's thesis, Department of Geology and Geography, Kansas State ‘University, Manhattan, Twiss, P.C., E, Suess, and R. M. Smith 1969." Morphological Classification of Grass Phytoliths. Proceedings of the Soil Science Society of America 33:109-115. Madison, Wisconsin. Wilding, L. P., and L. R. Drees 1971 ' Biogenic Opal in Ohio Soils. Proceedings of the Soil Science Society of America 35:1004-1010. Madison, 13-119.104 AMERICAN ANTIQUITY {Vol $5, No. 1, 1990} 1973. Scanning Electron Microscopy of Opaque Opaline Forms Isolated from Forest Soils in Ohio. Pro- ceedings of the Soil Science Society of America 37:647-650. Madison, Wisconsin. 1974 Contributions of Forest Opal and Associated Crystalline Phases of Fine Silt and Clay Fractions of Soils. Clays and Clay Minerals 22:295-306. Received July 6, 1988; accepted September 13, 1988 IDENTIFICATION OF THE SPECIES OF ORIGIN OF RESIDUAL BLOOD ON LITHIC MATERIAL D. C. Hyland, J. M. Tersak, J. M. Adovasio, and M. I. Siegel The examination of stone-tool edges for blood-antigen residue is a relatively new technique in the archaeological analysis of lithic material. To date, a number of different methods have been employed t0 determine the species of origin of residual blood, such as Ouchterlony double-diffusion (Ouchterlony 1968) and radioimmunoassay (RIAs) (Lowenstein 1985, 1986). These techniques have been of limited use due to problemes of sensitivity, cost, ‘and applicability to archaeological fetd conditions. Enzyme immunoassays (EIAs) utilizing a nitrocellulose pro- tein-binding membrane have shown that it is possible 10 retain both specificity and sensitivity in a technique that can endure the rigors of archaeological field work and the vicissitudes of differential preservation (Tersak and Hyland 1988). However, due to cross-reactivity, species identification of blood residue using immunological techniques can be problematic when itis necessary to distinguish individual species within a single genus or family. Recent work has shown that careful refinement of antisera can eliminate cross-reactivity, thereby increasing specificity (Berkeley Antibody Company, personal communication 1987}. This refinement is accomplished by ‘absorbing any antibodies from the anuisera that may cross-react with antegenic sites held in common by closely related species. The ramifications of the successful implementation of this technique are discussed in terms of Paleoindian artifact function as well ax paleoeconomic, paleoenvironmentat, and pateadietary reconstruction ustag 1 case study from the Shoop site in central Pennsylvania. Elexamen del filo de implementos fiticos para la biisqueda de residuos cle antigenos sangutneos es una técnica relativamente nueva en el estudio arqueoldgico de material litico. Hasta esta fecha varios métodas se han empleado ‘para determinar las especies de origen de sangre residual. tales como doble-difusiin Ouchterlonea (Ouchterlony 1968) y radioimmunoassays (RIAs) (Lowenstein 1985, 1986). Estas técntcas han tenido uso limitado debido a problemas de sensibilidad, costo, y aplicabilidad bajo condiciones de terreno arqueolégico. La enzima immu- noassays (EAs), utilizando wna membrana nitrocelulosa que ata proteinas ha demostrado que es posible retener ‘especifcidad y sensibilidad en una técnica que puede aguantar tanto los rigores de trabajo arqueoldgico en terreno ‘como las visicitudes de diferentes grados de preservaci6n. (Tersak and Hyland 1988). Sin embargo, debido a la reactividad entrecruzada, la identificacton de especies atravez de residuos de sangre usando técnicas imunologicas puede ser problematica cuando se hace necesario identificar especies individuales dentro de wn genero o familia Recientemente se ha demostrado que el cuidadoso refinamiento de antisera puede eliminar reactividad entrecruzada ‘yaumentar especificidad. (Berkeley Antibody Company, comunicacidn personal 1987). Este refinamiento se logra ‘absorbiendo cualquier anticuerpo del antisera que pueden tener una reacciOn cruzada con sitios antigénicos que tienen en comin especies estrechamente relacionadas. Las ramificaciones del exitoso uso de esta técnica se discuten en relaci6n a la funcidn de artefactos paleoindios y también en términos de reconstruccién paleoeconémica, paleomedioambiental y paleodietética usando como ejemplo un estudio del sitio Shoop de Pennsylvania. Despite the fact that the theoretical strides American archaeology has taken in the past 40 years have received the bulk of the attention in histories of the field (¢.g., Willey and Sabloff 1980), it is D.C Hyland, IM. Tersak, J. M. Adovasio, and M. I. Siegel, Department of Anthropology, University of Pittsburgh, Pitsburgh, PA 15260 American Antiquity, $5(1), 190, pp, 104-112. Copyright © 1990 by the Society for American Archaeology