PHYTOLOGIA An international journal to expedite plant systematic phytogeographical and ecological publication www.phytologia.org Vol. 91, No. 1, pp 1-188 April 2009 PHYTOLOGIA (ISSN 00319430) Phytologia, a journal for rapid publication in plant systematics, phytogeography and vegetation ecology, is published three times a year. Managing Editor Executive Editor micro-Molecular, Phytochemical General systematics and Multivariate Systematics and evolution Robert P. Adams Billie L. Turner Baylor University University of Texas at Austin Baylor-Gruver Lab Austin, TX 78705 Gruver, TX 79040 billie@uts.cc.utexas.edu Robert_ Adams@baylor.edu Associate Editors Nomenclature General Systematics Guy Nesom A. Michael Powell 2925 Hartwood Drive Dept. of Biology Fort Worth, TX 76109 Sul Ross State University www.guynesom.com Alpine, TX, 79832 ampowell@sulross.edu Macro-Molecular and Ethnobotany Phylogenetic Systematics Martin Terry Andrea E. Schwarzbach Dept. of Biology Univ. of Texas at Brownsville Sul Ross State University Dept. of Biol. Sciences Alpine, TX, 79832 Brownsville, TX, 78520 mterry@sulross.edu andrea.schwarzbach@utb.edu Secretary-Treasurer - Subscriptions Robert P. Adams Phytologia, Box 727, Gruver, TX 79040 Robert Adams@baylor.edu Copyright 2008 Phytologia. Texensis Publishing, Gruver, TX Phytologia (April 2009) 91(1) l Phytologia Contents M. Woods and J. Key. The genus Rhynchosia (Fabaceae) in Alabama.. D. S. Seigler and J. E. Ebinger. New combinations in the genus J Bemecmia (Pabaceac: (Mimosoideae)s.i10. «ils dees iis otled Sad deade Xs 26 R. P. Adams. The leaf essential oil of J. maritima R. P. Adams compared with J. horizontalis, J. scopulorum and J. virginiana oils...31 R. P. Adams. Geographic variation and systematics of Juniperus phoenicea from Madeira and the Canary Islands: Analyses of leaf NAS oc dp at crate tid Mercier Oa aces nine nae aac pee 40 J. E. Averett. Schraderanthus, A new genus of Solanaceae............. 54 G. L. Nesom. Point of view: Authors' initials in scientific names with RaDESEE URGHORICIOS ( 5, cic bicd th. Saei seine edb ace BEM cae Sta ie Sark ae A tg 62 G. L. Nesom. Maytenus phyllanthoides (Celastraceae) in Louisiana, ft MAKEMOtes On taxonomy Of fhe SPECIES. 3/2. :...038.0..skeease~ ees scneoeewe! 64 J. R. Singhurst, D. J. Rosen and W. C. Holmes. Two new additions to MRMRGEEE II SOA Of U ORAS 56 5 5st cian deneacbae acs sane sbaanwtneenteucees 69 « J. R. Singhurst, K. M. Fleming, R. Loper, and W. C. Holmes. Romulea rosa inidacéae)Adventive in Texas.cnei eee. -.k. eS A 13: 4 Cover Photos: Morus murrayana by Dayle E. Saar and Euthamia caroliniana by Jason Singhurst. ee 0" NICAI tAR 2 Phytologia (April 2009) 91(1) B. L. Turner. Taxonomy of /Jva angustifolia and I. asperifolia - (ASTErACERE) i550. sas adnrne somceco eemeat aae tamer tne tna uot ke iaieaiecabe saa eee 1G. B. L. Turner. A new species of Trixis (Asteraceae: Mutisieae) from i JAaliSCOMMIENICO’ oo. posisciassotycan cucee ee RS eee nee caja see a oe ee 84 B. L. Turner. Carminatia papagayana (Asteraceae: Eupatorieae), A new Species from western Guerrero; MIEXICO) a sce 5 es dt 4 mm. dia.), or if small, with sharply serrate leaves; leaves and inflorescences usually with some hairs; lateral veins protruding above lower leaf surface. 5.Leaves, or at least some, with acute apices; blades elliptic, unlobed or with 2-4 lateral lobes. 6.Fruits medium (4-7 mm. dia.), usually in several-flowered corymbs; lower surface of leaves with small sunken pockets in axils of major veins; thorns few. Small to mid-sized tree. Floodplains, riverbottom forests, swamps. Panhandle and north Florida, south to north peninsula (Marion County); frequent. Early spring. [Crataegus paludosa Sarg.] GREEN HAW. Crataegus viridis L. 6.Fruits large (6-15 mm. dia.), solitary or usually so (if several together, usually not of the same corymb). 7.Lower surface of leaves with pockets lacking in vein axils; petioles distinct, the blade cuneate at base but not extended downward; leaves usually with 2-4 shallow sharply-serrate lobes; thorns few. Small to mid-size tree. Open woodlands, moist hammocks and slopes. Panhandle, locally eastward (to Alachua, Columbia, Levy counties); frequent. Spring. [Crataegus opima Beadle; Crataegus pulcherrima Ashe; Crataegus rober Beadle] SMOOTH HAW. Crataegus flava Ait. 7.Lower surface of leaves with prominent hair-filled sunken pockets in axils of major lateral veins (domatia); leaves unlobed or deeply but irregularly cut or lobed; petioles indistinct, changing gradually into cuneate-based blade. 8.Leaves 3-5 cm. long, serrate or crenate toward apex, glossy above, nearly glabrous below; thorns few. Shrub or small tree. Pond margins, creek banks. Mid- panhandle (Gadsden County), east to northeast Florida (Nassau County), south along east coast (to Volusia County); frequent. Early spring. [Crataegus luculenta Sarg.; Crataegus maloides Sarg.] Phytologia (April 2009) 91(1) MAY HAW. Crataegus aestivalis (Walt.) Torr. & Gray 8.Leaves 5-7 cm. long, entire or finely serrate-crenate toward apex, dull green above, densely rufous-tomentose below when young, persisting along veins when full- grown. 9.Leaves elliptic to broad-elliptic; lateral veins 5-9; pedicels glabrous; fruits 12-15 mm. dia. Small tree. Stream and river banks. Western panhandle (n.e. Escambia County: Century); rare. Spring. APPLE HAW. Crataegus opaca Hook. & Arn. 9.Leaves broad-elliptic to obovate; lateral veins 3-5; pedicels rufous-tomentose; fruits 10-12 mm. dia. Small tree. River bottoms. Central panhandle (Jackson, Gadsden counties); rare. [Crataegus aestivalis, misapplied] RUFOUS MAY HAW. Crataegus rufula Sarg. 5.Leaves with rounded apices, or a few acute; blades oblanceolate to spatulate, without lobes (or some leaves irregularly lobed on vigorous shoots of C. michauxii); fruits large (6-12 mm. dia.). 10.Leaves glossy green above, without glands on petioles or margins, without pockets (domatia) in vein axils; thorns usually very prominent (3-4 cm. long). Small tree. Floodplain forests, moist hammocks. North Florida (excl. w. panhandle), south into north peninsula (to Clay, Levy counties); frequent. Spring. [Crataegus pyracanthoides Beadle] COCKSPUR HAW. Crataegus crus-galli L. 10.Leaves uniformly light green above and below; petioles and margins with dark button-like glands; lower leaf surface with proximal vein axils sunken and hairy, forming small pockets (domatia); thorns numerous but small (1-2 cm. long). Spring. An apomictic complex, of many named forms, only two of which are here given recognition. SUMMER HAW, YELLOW HAW. Crataegus michauxii Pers. a.Leaves broadly spatulate to obovate; branches variously spreading, not weeping. Shrub or small to mid-sized Phytologia (April 2009) 91(1) 25 tree. Dry to moist woodlands, fencerows, open pastures. North Florida, south to mid-peninsula (Highlands County); common. [Crataegus audens Beadle; Crataegus egregia Beadle; Crataegus flava, misapplied; Crataegus floridana Sarg.; Crataegus galbana Beadle; Crataegus leonensis Palmer; Crataegus lepida Beadle; Crataegus ravenelii Sarg.; Crataegus visenda Beadle] SUMMER HAW (typical). var. michauxii a.Leaves narrowly spatulate (except on vigorous shoots, when sometimes broader); branches and_ branchlets prominently weeping (pendent). Small tree. Dry sandhills. Western half of panhandle (Escambia to Calhoun counties); infrequent and local. [Crataegus lacrimata Small] WEEPING HAW. _. var. lacrimata (Small) D. B. Ward *This paper is a continuation of a series begun in 1977. The "amplified key" format employed here is designed to present in compact form the basic morphological framework of a conventional dichotomous key, as well as data on habitat, range, and frequency. Amplified keys are being prepared for all genera of the Florida vascular flora; the present series is restricted to genera where a new combination is required or a special situation merits extended discussion. 26 Phytologia (April 2009) 91(1) NEW COMBINATIONS IN THE GENUS SENEGALIA (FABACEAE: MIMOSOIDEAE) David S. Seigler Department of Plant Biology, University of Illinois, Urbana, Illinois 61801, U.S.A. email: seigler@life.uiuc.edu John E. Ebinger Emeritus Professor of Botany, Eastern Illinois University, Charleston, Illinois 61920, U.S.A. email: jeebinger@eiu.edu ABSTRACT Morphological and genetic differences separating the subgenera of Acacia s./. and molecular evidence that the genus Acacia s.l. is polyphyletic necessitate transfer of the following taxa from Acacia subgenus Aculeiferum Vassal to Senegalia, resulting in 10 new combinations in the genus Senegalia: S. grandisiliqua (Benth.) Seigler & Ebinger, S. guarensis (L. Cardenas & F. Garcia) Seigler & Ebinger, S. laeta (R. Br. ex Benth.) Seigler & Ebinger, S. Jowei (L. Rico) Seigler & Ebinger, S. polyacantha (Willd.) Seigler & Ebinger, S. riograndensis (Atahuachi & L. Rico) Seigler & Ebinger, and S. skleroxyla (Tussac) Seigler & Ebinger. Two new combination and new status changes are necessary: S. latifoliola (Kuntze) Seigler & Ebinger, comb. et stat. nov. and S. rhytidocarpa (L. Rico) Seigler & Ebinger, comb. et stat. nov. A lectotypification is made for Acacia grandisiliqua Benth. One new name was required: S. stenocarpa Seigler & Ebinger, nom. nov. Phytologia 91(1): 26-30,( April, 2009). KEY WORDS: Acacia sensu lato, Fabaceae, Mimosoideae, Senegalia. Morphological and _ genetic differences separating the subgenera of Acacia s./. and molecular evidence that the genus Acacia s.l. is polyphyletic necessitate recognition of segregate genera and Phytologia (April 2009) 91(1) 27 transfer of many Acacia species to these genera. A large number of species of Acacia subgenus Aculeiferum Vassal must be referred to Senegalia Rafinesque. Taxonomic Changes For the following 10 taxa, this results in new combinations, changes in status and a new name in the genus Senegalia: 1. SENEGALIA GRANDISILIQUA (Benth.) Seigler & Ebinger, comb. nov. Basionym: Acacia grandisiliqua Benth. London J. Bot. 1: 518. 1842. Bentham equated this taxon with Mimosa grandisiliqua Vell., Fl. flumin. 11: (t. 37). 1790 [1831]. nom. nud. and made a nom. nov. based on the type specimens below. — TYPE: BRAZIL. BAHIA: C. F. P. von Martius 1098 [lectotype, designated here: K (photo F)]; [paratypes: Lushnath 145 (K, MO); C. F. P. von Martius 1104 (K?)]. 2. SENEGALIA GUARENSIS (L. Cardenas & F. Garcia) Seigler & Ebinger, comb. nov. Basionym: Acacia guarensis L. Cardenas & F. Garcia, Ernstia 10: 146. 2000.— TYPE: VENEZUELA: Territorio Federal Amazonas. Cuenca del Rio Manapiare, selva alta, a media hora de camino desde el poblado de Guara en direccion SE, en la pica hacia el Cafio Garrafon, alt. 140 m, 5° 15’ N, 66° 03’ W, 28 Jan 1977, O. Huber 429/3435 (holotype: VEN 307571). 3. SENEGALIA LAETA (R. Br. ex Benth.) Seigler & Ebinger, comb. nov. Basionym: Acacia laeta R. Br. ex Benth., London J. Bot. 1: 508. 1842.— TYPE: ETHIOPIA: prope montes Tarnta, H. Salt 82 (holotype: BM; isotypes: MO, NY). 4. SENEGALIA LATIFOLIOLA (Kuntze) Seigler & Ebinger, comb. et stat. nov. Basionym: Acacia riparia Kunth var. latifoliola Kuntze, Revis. gen. pl. 3(2): 47. 1898.- TYPE: BRAZIL. MATO GROSSO: O. Kuntze s.n. (holotype: NY).\ 28 Phytologia (April 2009) 91(1) 5. SENEGALIA LOWE I (L. Rico) Seigler & Ebinger, comb. nov. Basionym: Acacia lowei L. Rico, Amer. Sp. Acacia 114. 2007. — TYPE: Probably from Brazil, but based on a specimen cultivated in Madeira; [lectotype, Rico-Arce (2007): original drawing for plate 3366 (M. Young, K-library)]. 6. SENEGALIA POLYACANTHA (Willd.) Seigler & Ebinger, comb. nov. Basionym: Acacia polyacantha Willd., Sp. pl. 4: 1079. 1806.— TYPE: No type cited, habitat in India orientali, Roxburgh s.n. [holotype: B-Willd.; isotype: (fragment K)]. 7. SENEGALIA RHYTIDOCARPA (L. Rico) Seigler & Ebinger comb. et stat. nov. Basionym: Acacia polyphylla DC. var. rhytidocarpa L. Rico, Anales Jard. Bot. Madrid 63: 28. fig. 1. 2006. — TYPE: BOLIVIA. BENI: Provincia Ballivian, km 35 carretera Yucumo-Rurrenabaque, Colegio Técnico Agropecuario, rio Colorado, 67° 05’ W, 14° 50’ S, D. N. Smith et al. 13586 (holotype: LPB; isotypes: G, K, MO). 8. SENEGALIA RIOGRANDENSIS (Atahuachi & L. Rico) Seigler & Ebinger, comb. nov. Basionym: Acacia riograndensis Atahuachi & L. Rico. Kew Bull. 62: 605. 2007.— TYPE: BOLIVIA. COCHABAMBA: Provincia Campero, Pasorapa, en la bajada de Buena Vista hacia el Rio Grande, 27 Dec 2004, /. R. I. Wood, M. Atahuachi & M. Mercado 21251 (holotype: BOLV; isotypes: K, LPB). 9. SENEGALIA SKLEROXYLA (Tussac) Seigler & Ebinger, comb. nov. Basionym: Acacia skleroxyla Tussac, Fl. Antill. 1: 146. (pl. 21). 1808 [1808-1813].— TYPE: Antilles. Santo Domingo [holotype: pl. 21 from Tussac (1808); isotype: K (Rico-Arce 2007)]. Phytologia (April 2009) 91(1) 29 10. SENEGALIA STENOCARPA Seigler & Ebinger, nom. nov. Basionym: Acacia stenocarpa Malme, Ark. Bot. 23A(13): 46. 1931. nom. illeg. non Richard (1847).— TYPE: BRAZIL. MATO GROSSO: Corumba, in silva satis clara regionis calcariae, 19 Dec 1902, G. O. A. Malme 2731 [lectotype, Seigler et al. (2006): S) [paratype: Malme 2731a(S)]. ACKNOWLEDGEMENTS The authors wish to thank several colleagues for advice concerning questions of nomenclature and general taxonomic advice. Among these are: Fred Barrie, J. Lee Crane, K. N. Gandhi, Bruce R. Maslin, Gordon Tucker, Paul Wilson, and James Zarucchi. The views and conclusions (and errors) in this manuscript are our own and do not necessarily reflect any of their judgments. We wish to acknowledge support by the National Science Foundation (NSF DEB 04-15803) and by the American Philosophical Society (1992). LITERATURE CITED Bentham, G. 1842. Notes on Mimoseae, with a synopsis of species, London Journal of Botany 1: 318-392. 494-528. Bentham, G. 1875. Revision of the suborder Mimoseae. Transactions of the Linnaean Society of London 30: 335-664. Colla, A. 1834. Herbarium Pedemontanum. 2: Torino. Ex Typis regiis. pp. 1-557. Richard, A. 1847 [1848]. Tentamen Florae Abyssinicae 1: Paris. 472 pp. Rico-Arce, L. 2007. American Species of Acacia. A Checklist and Synopsis of American species of Acacia (Leguminosae: Mimosoideae), CONABIO, Mexico City. 207 pp. 30 Phytologia (April 2009) 91(1) Seigler, D. S., J. E. Ebinger, and J. T. Miller. 2006. The genus Senegalia from the New World. Phytologia 87: 38-93. Tussac, F. R. de. 1808. Flores des Antilles, ou Histoire Generale. Vol. 1. Apud Autorem et F. Schoell, Paris. 1-198 pp. Phytologia (April 2009) 91(1) 3] THE LEAF ESSENTIAL OIL OF JUNIPERUS MARITIMA R. P. ADAMS COMPARED WITH J. HORIZONTALIS, J. SCOPULORUM AND J. VIRGINIANA OILS Robert P. Adams Biology Department, Baylor University, Box 727, Gruver, TX, 79040 Robert_ Adams@baylor.edu ABSTRACT This is first report on the composition of the leaf essential oil of J. maritima R. P. Adams, a new juniper species from the Pacific northwest USA. The volatile leaf oil of J. maritima is dominated by elemicin (20.2%), sabinene (20.0%), limonene (11.7%) and 8-a- acetoxyelemol (6.1%) with moderate amounts of safrole (3.8%), pregeierene B (3.1%) and terpinen-4-ol (1.8%). The leaf oils of J. horizontalis, J. scopulorum and J. virginiana var. virginiana were re- analyzed and compared with the oil of J. maritima. Each of the four species has a distinct oil composition reflecting their specific status. Phytologia 91(1):31-39, (April, 2009). KEY WORDS: Juniperus maritima, J. horizontalis, J. scopulorum, J. virginiana var. virginiana, Cupressaceae, essential oil composition, elemicin, sabinene, limonene, 8-a-acetoxyelemol. Adams (1983) examined geographic variation in leaf terpenoids throughout the range of Juniperus scopulorum Sarg. and found that plants from the Puget Sound area of northwestern North America showed considerable differences in their leaf terpenoids compared with typical J. scopulorum plants in the Rocky Mountains. Recent DNA sequencing (Schwarzbach et al, in prep.) found that the Puget Sound plants were more related to J. virginiana L. than J. scopulorum. As part of a continuing study of the genus Juniperus (Adams, 2004), additional plants were collected from the Puget Sound area and SNPs (Single Nucleotide Polymorphisms) were examined (Adams, 2007). That study (Adams, 2007) revealed (Fig. 1) that the junipers in the Puget Sound area also differed in their nrDNA SNPs. Based on a combination of SNPs, terpenoids, morphology and ecology, 32 Phytologia (April 2009) 91(1) Adams (2007) recognized the junipers of Puget Sound as a new species, J. maritima R. P. Adams, the seaside juniper. Although Adams (1983) reported on multivariate differences in the oils from Puget Sound plants (now J. maritima), no information was published on the oil composition. Because J. maritima is closely related to J. horizontalis, J. scopulorum and J. virginiana, these leaf oil compositions are included in this report. The leaf oil compositions of J. horizontalis, J. scopulorum and J. virginiana have been recently published (Adams, 2000). PCO 18 nrDNA SNPs J. scopulorum J. virginiana J. maritima 3 (6%) Figure 1. Principal coordinate Ordination (PCO) based on 18 SNPs. Note that J. maritima is genetically differentiated from J. scopulorum and J. virginiana. Modified from Adams (2007). Phytologia (April 2009) 91(1) 33 The purpose of this report is to present analysis of the leaf essential oil of J. maritima and compare the oil with the leaf oils of the most closely species: J. horizontalis, J. scopulorum, and J. virginiana. MATERIAL AND METHODS Plant material - Specimens used in this study : J. maritima: Brentwood Bay, Vancouver Isl., BC, Adams 11056-58, Cowichan Bay, Vancouver Isl., BC, Adams //06/-63, Yellow Point, Vancouver Isl., BC, Adams /1064, Lesqueti Isl., BC, Adams //065-66, Friday Harbor, San Juan Isl., WA, Adams /1067-68, Whidbey Isl., Cranberry L., WA, Adams 1/1075, Fidalgo Isl., Washington State Park, WA, Adams //076, Skagit Isl., WA, Adams /1077-78; J. horizontalis: Saskatchewan River bank, Saskatoon, Saskatchewan, Adams 1651-1660; J. scopulorum: w bank of Animas River, Durango, CO, Adams 2010-2024; and J. virginiana, 16 km e of Dulles Airport, Washington, DC, Adams 2409- 2423. Voucher specimens are deposited at the Herbarium, Baylor University (BAYLU). Isolation of Oils - Fresh leaves (200 g) were steam distilled for 2 h using a circulatory Clevenger-type apparatus (Adams, 1991). The oil samples were concentrated (ether trap removed) with nitrogen and the samples stored at -20°C until analyzed. The extracted leaves were oven dried (100°C, 48 h) for determination of oil yields. Chemical Analyses - Oils from 10-15 trees of each of the taxa were analyzed and average values are reported. The oils were analyzed on a HP5971 MSD mass spectrometer, scan time 1/ sec., directly coupled to a HP 5890 gas chromatograph, using a J & W DB-5, 0.26 mm x 30 m, 0.25 micron coating thickness, fused silica capillary column (see 5 for operating details). Identifications were made by library searches of our volatile oil library (Adams, 2006), using the HP Chemstation library search routines, coupled with retention time data of authentic reference compounds. Quantitation was by FID on an HP 5890 gas chromatograph using a J & W DB-5, 0.26 mm x 30 m, 0.25 micron coating thickness, fused silica capillary column using the HP Chemstation software. 34 Phytologia (April 2009) 91(1) Data Analysis - Terpenoids (as per cent total oil) were coded and compared among the species by the Gower metric (1971). Principal coordinate analysis was performed by factoring the associational matrix using the formulation of Gower (1966) and Veldman (1967). RESULTS AND DISCUSSION The volatile leaf oil of J. maritima is dominated (table 1) by elemicin (20.2%), sabinene (20.0%), limonene (11.7%) and 8-a- acetoxyelemol (6.1%) with moderate amounts of safrole (3.8%), pregeijerene B (3.1%) and terpinen-4-ol (1.8%). Several components are found only in J. maritima: isoamyl isovalerate (t), naphthalene (0.5), (2E,4Z)-decadienal (t), a-cubebene (t), a-humulene (t), B- bisabolene (0.3), zonarene (t), C;;O0H (AI 1586) (0.5) and cedrol (0.1). Most of these unique components are in trace amounts (less than 0.05%), and might be present in the other three juniper species in this study. It is interesting that cedrol is present as it is rare in the leaf oils of Juniperus in the western hemisphere (Adams, 2004). It is unusual that the leaf oil of J. maritima contains such large quantities of non- terpenoid (phenolic) compounds (elemicin, safrole). The overall pattern of variation was determined by computing similarity measures among the taxa and subjecting the associational matrix to principal coordinates analysis (PCO). Figure 2 shows the PCO ordination based on the terpenoids. Each of the sticks represents 10-15 individuals: J. horizontalis (10); J. maritima (15); J. scopulorum (15) and J. virginiana (15). From this analysis, each oil appears distinct. However, the oil of J. maritima appears most similar to the oil of J. virginiana. It is interesting that J. maritima is separated by 4 SNPs from J. virginiana and 5 SNPs from J. scopulorum (Fig. 1) similar to the pattern of leaf oils (Fig. 2). Phytologia (April 2009) 91(1) 35 PCO Terpenoids J. virginiana J. scopulorum J. maritima J. horizontalis Figure 2. Principal coordinate ordination (PCO) utilizing terpenoids. ACKNOWLEDGMENTS Thanks for field assistance from: Steve Erickson, Whidbey and Skagit Island, WA; Eugene Kozloff, San Juan Island; Peter Dederich, NPS, English Camp, San Juan Island, WA; Ted Smith, Deception Pass State Park, Whidbey Island, WA; Richard Hebda, Adolf Ceska, and John Pinder-Moss, Royal British Columbia Museum (V); Lance Goldy, Yellow Point Resort, BC; and Drew Chapman and Wade Calder, Lesqueti Island Ecological Reserve, BC. Thanks to Tonya Yanke for lab assistance. 36 Phytologia (April 2009) 91(1) LITERATURE CITED Adams, R. P.. 1983. Infraspecific terpenoid variation in Juniperus scopulorum: Evidence for Pleistocene refugia and recolonization in western North America. Taxon 32: 30-46. Adams, R. P. 2004. Junipers of the world: The genus Juniperus. Trafford Publ., Vancouver, B.C., Canada. Adams, R. P. 2007. Juniperus maritima, the seaside juniper, a new species from Puget Sound, North America. Phytologia 89: 263-283. Adams, R. P. 2000. Systematics of smooth leaf margin Juniperus of the western hemisphere based on leaf essential oils and RAPD DNA fingerprinting. Biochem. Syst. Ecol. 28: 149-162. Adams, R. P. 1991. Cedar wood oil - analysis and properties. In Modern Methods of Plant Analysis: Oils and Waxes. H. F. Linskins and J. F. Jackson, Eds., pp. 159 - 173, Springler-Verlag, Berlin, Germany. Adams, R. P. 2006. Identification of Essential Oils Components by Gas Chromatography/ Mass Spectrometry, 4th Ed. Allured Publ. Corp., Carol Stream, IL. Gower, J. C. 1971. A general coefficient of similarity and some of its properties. Biometrics 27: 857-874. Gower, J. C. 1966. Some distance properties of latent root and vector methods used in multivariate analysis. Biometrika 53: 326-338. Veldman, D. J. 1967. Fortran programming for the behavioral sciences. Holt, Rinehart and Winston Publ., NY. Phytologia (April 2009) 91(1) a Table I. Compositions of the leaf oils of J. maritima, (marit), J. virginiana (virg), J. scopulorum (scop) and J. horizontalis (horiz). AI Compound marit virg scop horiz 921 tricyclene . t t - 924 a-thujene 0.6 0.2 i 0.6 932 a-pinene 0.6 1.4 4.7 Li 945 a-fenchene - - t t 946 camphene - 0.1 0.1 t 969 sabinene 20.0 6.7 46.3 3712 974 -pinene t t 0.2 0.4 988 myrcene 0.9 0.7 13 2.8 1001 6-2-carene - t - 0.1 1002 a -phellandrene - . 0.1 t 1008 6-3-carene . 0.1 0.1 0.4 1014 a -terpinene 0.7 0.3 a 0.6 1020 p-cymene t 0.1 0.5 0.4 1024 limonene 11.7 193 5.4 329 1025 B-phellandrene - - 1.0 t 1044 (E)-B-ocimene - t 0.1 OZ 1054 y-terpinene 13 0.4 1.9 V2 1065 cis-sabinene hydrate 0.5 0.3 1.4 1.3 1086 terpinolene 0.7 0.5 0.8 0.7 1187 2-nonanone - - 0.2 - 1095 linalool 0.1 4.0 0.3 0.3 1098 trans-sabinene hydrate 0.2 - 1.0 0.5 1100 n-nonanal 0.1 0.2 - t 1102 isoamyl-isovalerate t - - - 1112 trans-thujone (= B-thujone) = - - 0.1 0.1 1118 cis-p-menth-2-en-1-ol 0.3 0.4 0.4 0.4 1136 trans-p-menth-2-en-1-ol t - 0.2 0.2 1141 camphor - 4.0 0.2 0.2 1145 camphene hydrate - 0.2 0.1 ~ 1148 citronellal : t - . 1165 borneol 0.1 0.7 . - 1167 umbellulone - - . t 1066 coahuilensol - 0.6 - - 1174 terpinen-4-ol 1.8 1.4 5.8 3.9 38 1178 1189 1186 1195 1195 1207 1219 1223 1249 1253 1255 1257 1274 1285 1287 1292 1314 1513 1322 1345 1350 1356 1379 1387 1403 1417 1442 1448 1452 1461 1468 1475 1478 1480 1493 1493 1500 1505 naphthalene p-cymen-8-ol - a-terpineol methyl chavicol cis-piperitol - trans-piperitol . coahuilensol, methyl ether - citronellol piperitone - trans-sabinyl hydrate acetate - (4Z)-decenol t methyl! citronellate 0.2 pregeyerene B 31 safrole 3.8 bornyl acetate t (2E,4Z)-decadienal decadienol isomer* (2E,4E)-decadienal t methyl] geranate t a-cubebene t citronellyl acetate - eugenol geranyl acetate - B-cubebene t methyl eugenol - (E)-caryophyllene 6,9-guaiadiene - cis-muurola-3,5-diene - a-humulene t cis-cadina-1(6),4-diene - pinchotene acetate - trans-cadina-1(6),4-diene 0.5 y-muurolene - germacrene D - trans-muurola-4(14), 5-diene 1.0 epi-cubebol 0.5 a-muurolene 0.4 B-bisabolene 0.3 Phytologia (April 2009) 91(1) = t = ioe 08 0.2 t = = : 0.1 t t 0.1 0.1 0.4 : 2 2A 08 : 0.3 2 2 = t ws O39 Oi 0.2 0.1 t : 5.) e610 : 10.0 t t 4.0 0.7 0.5 : 0.1 ' 0.2 : : t = ~ 0.1 : : t = = t = = 52. “Ga : t 0.1 2 : 0.2 0.2 : : 0.1 : : 0.1 0.1 5 - : 2 t : 0.1 0.3 t : 0.1 : 2 0.2 i 0.1 0.8 Oi 300 1.0 Phytologia (April 2009) 91(1) [513 1513 1522 1528 1537 1549 1555 1359 1574 1586 1600 1607 1627 1630 1638 1638 1644 1649 1652 1653 1670 1685 1739 1792 1887 2055 2056 2298 y-cadinene cubebol 6-cadinene zonarene a-cadinene elemol elemicin germacrene B germacrene D-4-ol C,5OH, 43,207,161,222 cedrol B-oplopenone 1-epi-cubenol y-eudesmol epi-a-cadinol epi-a-muurolol a-muurolol B-eudesmol a-eudesmol a-cadinol bulnesol germacra-4(15),5,10(14)- trien-1-al oplopanone 8-a-acetoxyelemol oplopanonyl acetate abietatriene manool 4-epi-abietal ao AI = Arithmetic Index on DB-5 column. Values less than 0.05% are denoted as traces (t). Unidentified components less than 0.5% are not reported. Those compounds that appear to distinguish taxa are in boldface. 40 Phytologia (April 2009) 91(1) GEOGRAPHIC VARIATION AND SYSTEMATICS OF JUNIPERUS PHOENICEA L. FROM MADEIRA AND THE CANARY ISLANDS: ANALYSES OF LEAF VOLATILE OILS Robert P. Adams Biology Department, Baylor University, Box 727, Gruver, TX, 79040 Beatriz Rumeu and Manuel Nogales Island Ecology and Evolution Research Group (IPNA-CSIC), 38206 La Laguna, Tenerife, Canary Islands, Spain and Susana S. Fontinha Parque Natural da Madeira, CEM-UMa, Caminho do Meio 9064-512, Funchal, Madeira, Portugal ABSTRACT All of the oils of J. phoenicea from the Canary Islands and Madeira were very similar. The volatile leaf oils were dominated by a- pinene (57.3 - 76%) as was the oil from Morocco (65.4%). This is higher than in J. p. var. phoenicea, Spain (41.2%) or var. turbinata, Spain, (25.8%). The Madeira and Canary Island oils had moderate amounts of B-phellandrene (0.5 - 8.0%), myrcene (2.3 - 3.3%), a- terpinyl acetate (trace - 5.0%), (E)-caryophyllene (0.4 - 1.4%), and trans-totarol (0.1 - 2.1%). There is some differentiation in the oils from Madeira and the Canary Islands from populations in Spain and Morocco, but not enough to justify the recognition of J. p. subsp. canariensis at this time. Phytologia 91(1):40-53 (April, 2009). KEY WORDS: Juniperus phoenicea, Cupressaceae, Madeira Island, Canary Islands, leaf essential oils, a-pinene, myrcene, B-phellandrene. Juniperus phoenicea L. of the Mediterranean has red seed cones (berries) and is the only serrate leaf margined juniper in section Sabina in the eastern hemisphere (Adams, 2008). Gaussen (1968) discussed several other infraspecific taxa: var. canariensis (Guyot & Mathou) Rivas-Martinez et al., of the Canary Islands, var. /ycia (L.) Gaussen, Phytologia (April 2009) 91(1) 4] France littoral zone, var. mollis M & W, Morocco, and _ var. megalocarpa Maire, dunes near Mogador, Morocco. Adams et al. (1996) examined leaf terpenoids of J. phoenicea var. phoenicea, Greece and Spain, J. p. var. turbinata (Guss.) Parl. (=var. oophora Kunze), Tarifa Sand Dunes, Spain and J. p. subsp. eu-mediterranea, west of Setubal, Portugal. Adams et al. (1996) concluded that J. p. var. turbinata is conspecific with J. p. subsp. eu-mediterranea. There are a number of older literature reports on analyses of the leaf volatile oil of J. phoenicea and these are reviewed in Adams et al. (1996). The Adams et al. (1996) study was followed up using RAPDs (Adams et al., 2002). Figure 1 shows the PCO based on 119 RAPD bands. Note that eu-mediterranea and v. turbinata form a cluster (lower left). However, the plants from Tenerife, Canary Islands (cf. v. canariensis, fig. 1) cluster closely with plants from Nea Epidavios, Greece! This study confirmed the previous terpene analyses (Adams, et al., 1996) that subsp. ew-mediterranea and v. turbinata are conspecific. The plants from Corsica Island and Delphi Greece formed a separate group. PCO 3 (14%) 119 RAPD bands fea = v. canariensis, Canary Islands © = v. eu-mediterranea, Portugal & =v. phoenicea, El Penon, Spain > = v. turbinata, Tarifa, Spain * = Corsica Island +x = Delphi, Greece w= Nea Epidavios, Greece 2 (14%) 1 (27%) Figure 1. PCO based on 119 RAPD bands ordinating various taxa of J. Phoenicea. 42 Phytologia (April 2009) 91(1) Most recently, Adams et al. (2006) analyzed RAPDs from J. phoenicea from sand and rock areas in Morocco and compared these populations with plants from Tenerife, Canary Islands and var. turbinata, Tarifa sand dunes, Spain. PCO ordination (fig. 2) shows that 41% of the variance in the RAPDs was due to the differences between var. phoenicea (Spain) and the Morocco, Tenerife and var. turbinata populations. The Tenerife population accounted for about 14% of the variance (fig. 2). Although, the Canary Island plants are loosely associated with var. turbinata, they generally have large, round berries (seed cones), not turbinate shaped. 3(14%) PCO 111 RAPD bands J. p. var. phoenicea Spain, 720m J. p. var. turbinata Tarifa sand, 30 m 2(20%) Morocco, 940 m, rock 1(41%) Canary Islands, 150 m, lava Figure 2. PCO ordination of J. phoenicea populations based on 111 RAPD bands. The purpose of this study was to report on the volatile leaf oil compositions of populations of J. phoenicea from several islands in the Canary archipelago and Madeira, and to contrast these oils with J. p. var. phoenicea (Iberian Peninsula, Spain) and var. turbinata (Tarifa Phytologia (April 2009) 91(1) 43 sand dunes, Iberian Peninsula, Spain) oils. The distribution of J. phoenicea in Madeira and the Canary Islands is shown in figure 3. Porto Santo ° Island Oy, Madeira Island J. phoenicea Lanzarote O La vg Tenerife Palma af L Fuerteventura La El Hierro Gomera Gran Canaria Sahara Canary Islands Figure 3. Distribution of J. phoenicea in Madeira and Canary Islands. MATERIALS AND METHODS Plant material - J. phoenicea Madeira Island: 32° 48.822'N, 16° 52.627'W, ca 100 m, R. P. Adams 11502, 11503, cultivated at Botanic Garden in Funchal, ex Porto de la Cruz, 32° 39.08'N, 16° 47.14'W, ca 100 m, R. P. Adams 11503; Canary Islands: Tenerife, volcanic rock, ca. 150 m, R. P. Adams 8147-8149, La Palma Island, Santa Lucia, loose volcanic pumice, 28° 44.250'N, 17° 44.198'W, 283 m, R. P. Adams 11514-11516, La Gomera Island, volcanic rock, 28° 11.358'N, 17° 12.320'W, 370 m, R. P. Adams 11528-115230; Spain, limestone soil, 25 km e. Guadahortuna, 720 m, El Penon, R. P. Adams, 7077-7079; Morocco, red clay, 20 km sse Marrakech, 31° 21.033'N, 07° 45.893'W, 940 m, R. P. Adams 9408-9410; Spain, J. phoenicea var. turbinata: 44 Phytologia (April 2009) 91(1) Tarifa sand dunes, 15 km w. of Tarifa, 30 m, 36° 04.996'N, 5° 42.104' W, R. P. Adams, 7202-7204. Voucher specimens are deposited at the Herbarium, Baylor University (BAYLU). Isolation of Oils - Fresh leaves (200 g) were steam distilled for 2 h using a circulatory Clevenger-type apparatus (Adams, 1991). The oil samples were concentrated (ether trap removed) with nitrogen and the samples stored at -20°C until analyzed. The extracted leaves were oven dried (100°C, 48 h) for determination of oil yields. Chemical Analyses - Oils from 10-15 trees of each of the taxa were analyzed and average values are reported. The oils were analyzed on a HP5971 MSD mass spectrometer, scan time 1/ sec., directly coupled to a HP 5890 gas chromatograph, using a J & W DB-5, 0.26 mm x 30 m, 0.25 micron coating thickness, fused silica capillary column (see 5 for operating details). Identifications were made by library searches of our volatile oil library (Adams, 2006), using the HP Chemstation library search routines, coupled with retention time data of authentic reference compounds. Quantitation was by FID on an HP 5890 gas chromatograph using a J & W DB-5, 0.26 mm x 30 m, 0.25 micron coating thickness, fused silica capillary column using the HP Chemstation software. Data Analysis - Terpenoids (as per cent total oil) were coded and compared among the species by the Gower metric (1971). Principal coordinate analysis was performed by factoring the associational matrix using the formulation of Gower (1966) and Veldman (1967). RESULTS AND DISCUSSION All of the oils from the Canary Islands and Madeira were very similar (table 1). The volatile leaf oils were dominated by a-pinene (57.3 - 76%) as was the oil from Morocco (65.4%). o-pinene was higher in concentration in than in J. p. var. phoenicea, Spain (41.2%) or var. turbinata, Spain, (25.8%). The Madeira and Canary Island oils had moderate amounts of B-phellandrene (0.5 - 8.0%), myrcene (2.3 - 3.3%), a-terpinyl acetate (trace - 5.0%), (E)-caryophyllene (0.4 - 1.4%), and trans-totarol (0.1 - 2.1%). Phytologia (April 2009) 91(1) 45 The oil from Morocco was the only oil with camphor (1.3%, table 1). The oil of J. p. var. phoenicea, Spain, contained a large concentration of manoy] oxide (22.0%). The oil of J. p. var. turbinata, Spain, contained large amounts of fB-phellandrene (31.5%) and a- terpinyl acetate (13.1%) along with the smallest amount of a-pinene (25.8%). Only two compounds seem to separate the oils of Madeira and Canary Islands from continental oils: (E)-2-hexenal and (Z)-3-hexenol (table 1). However, these very volatile components are easily lost during transport and distillation, so the lack of these compounds in the oils from Morocco and Spain (table 1) may not be so significant. The J. phoenicea oil from Madeira shows differentiation from the Canary Islands in having higher concentrations of B-phellandrene (8.0%), linalool (1.0%), a-terpinyl acetate (5.0%) and a-eudesmol (0.9%, vs. absent in the Canary Island oils, table 1). In general, these compounds point to a similarity to the oil of J. p. var. phoenicea from Spain. To better understand the similarities among the oils, similarity measures were computed and the matrix of associations was factored. Eigenroots were extracted and accounted for 31.08, 19.50, 18.77, and 13.0% of the variance among the seven samples. The eigenroots appeared to asymptote after the fourth eigenroot, implying that five groups may be present. Principal Coordinate Ordination (PCO) of the samples (Fig. 3) shows that the oils from the Canary Islands (La Gomera, La Palma and Tenerife) are very similar (0.77 - 0.84). The next most similar oil is from Madeira (0.73 to La Palma). The Canary Islands oils are then linked to Morocco (0.70). Juniperus phoenicea var. turbinata (Tarifa sand dunes, Spain) are the least similar and link to Madeira (0.60) just smaller than the link of J. p. var. phoenicea, Spain to Madeira (0.64). There is certainly considerable variation in the volatile leaf oil compositions from various populations of J. phoenicea from the populations sampled in this study. It is not clear if there is sufficient differentiation in the Canary Islands to support the recognition of J. phoenicea subsp. canariensis at this time. 46 Phytologia (April 2009) 91(1) Geographic variation among the samples was further analyzed by plotting a minimum spanning network onto a geographic map. The 2(19%) PCO 50 terpenoids Morocco ‘La Palma var. phoenicea so he B4-[] o>. Tenerife - ; = Madeira * th La Gomera .-* '60 @» Var. turbinata Spain Figure 4. PCO ordination based on 50 terpenoids with the minimum spanning network super-imposed. samples from the Canary Islands are, geographically, the nearest neighbors and their oils high similarities reflect the co-differentiation and genetic isolation of the Canary Islands from Africa and Madeira (ig, 5): However, the linkage of the Canary Islands populations to Madeira is larger than its linkage to Africa (fig. 5). This may reflect more gene flow from north - south bird migrations (and seed cone dispersal) than from the east-west bird migrations to Morocco. Phytologia (April 2009) 91(1) 47 Alternatively, the linkage to Madeira may reflect co-evolution in similar climates of the Canary Islands and Madeira. J. phoenicea Minimum Spanning Network 50 terpenoids Madeira Lanzarote ~ o/ p Fuerteventura oO on oO) El Hierro Gomera Gran Canaria Western Canary Islands Sahara Figure 5. Minimum spanning network based on 50 terpenoids. ACKNOWLEDGMENTS Thanks to whose helped us collect in the islands (Félix M. Medina- La Palma, Angel Fernandez - La Gomera, Paulo Moniz - Madeira Island). This research was supported in part with funds from Baylor University and the project 80/2005 from the Organismo Autonomo de Parques Nacionales, Ministerio de Medio Ambiente of Spain. 48 Phytologia (April 2009) 91(1) LITERATURE CITED Adams R. P. 1975. Statistical character weighting and similarity stability. Brittonia 27: 305-316. Adams, R. P. 1991. Cedar wood oil - analysis and properties. In Modern Methods of Plant Analysis: Oils and Waxes. H. F. Linskins and J. F. Jackson, Eds., pp. 159 - 173, Springler-Verlag, Berlin, Germany. Adams, R. P. 2006. Identification of Essential Oils Components by Gas Chromatography/ Mass Spectrometry, 4th Ed. Allured Publ. Corp., Carol Stream, IL. Adams, R. P., 2008. Junipers of the world: The genus Juniperus. 2nd Ed., Trafford Publ., Vancouver, B.C., Canada. Adams, R. P., A. F. Barrero and A. Lara. 1996. Comparisons of the leaf essential oils of Juniperus phoenicea L., J. phoenicea subsp. eu-mediterranea Lebr. & Thiv. and J/. phoenicea var. turbinata (Guss.) Parl. J. Essent. Oil Res. 8: 367-371. Adams, R. P., S. Nguyen and N. Achak. 2006. Geographic variation in Juniperus phoenicea (Cupressaceae) from the Canary Islands, Morocco and Spain based on RAPDs analysis. Phytologia 88(3): 270-278. Adams, R. P., R. N. Pandey, Serge Rezzi and Joseph Casanova. 2002. Geographic variation in the Random Amplified Polymorphic DNAs (RAPDs) of Juniperus phoenicea, J. p. var. canariensis, J. p. subsp. eu-mediterranea, and J. p. var. turbinata. Biochem. Syst. Ecol. 30: 223-229. Gaussen, H. 1968. Les Cupressacees Fasc. X in Les Gymnospermes, Actuelles et Fossiles. Lab. Forest, Univ. Toulouse, France. Gower J. C., 1971. A general coefficient of similarity and some of its properties. Biometrics 27, 857-874. Gower J. C., 1966. Some distance properties of latent root and vector methods used in multivariate analysis. Biometrika 53, 326-338. Veldman D. J., 1967. Fortran programming for the behavioral sciences. 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LO v0 0°22 LO LO . cl 80 Gb : LO . LO £0 £0 Z0 £0 c0 v0 c0 LO LO c0 LO c0 €0 v0 LO LO LO LO 92 c0 Oc v0 v0 €0 LO c0 v0 c0 } ejyeuiginy uledS Odd0I0/y eJAWOS e] Cujeg e] ajoue] elope Z0 joulBhns9aj-sues} Le ,|012}0}-SUBL} y'0 |eyaiqe-ida-p v0 susipeyaige €0 susijejaige - joouew ; apixo |Aouew-¢ 1 -Ida : x29pixo |Aouew SUSIP-G} ‘(p1)g-esewidooesepues - .lOSewej-(49‘ZZ) Ol lounqoAys 9°0 [>| -Udtd} -(pL)OL‘S‘(SL)p-essews9b6 - josaujng - ,JOUIped-D 60 ,[OWsapne-p Z0 jousapne-g - jouaqno €0 jOjounnWw-p-Ida Z0 |Oulpeo-p-Ida - jowsapne-A v0 .022'70Z'191'6LL Eb ‘HOD punodwioy LEE? VLETS 8622 2802 GS0Z2 9502 6002 8L6L 8961 SLLZL 889L S89L OL9L ZS9L cS9OL 6v9l SV9L 8E9l 8e9l O€9L Gc9OL 54 Phytologia (April 2009) 91(1) SCHRADERANTHUS, A NEW GENUS OF SOLANACEAE John E. Averett Department of Biology, P.O. Box 8042 Georgia Southern University, Statesboro, GA 30460 U.S.A. averett@georgiasouthern.edu ABSTRACT Saracha viscosa Schrader, of Mexico and Central America, has been positioned in a variety of genera since it was first described, most notably Athenaea, Jaltomata and Leucophysalis. Arguments are presented for the exclusion of the taxon from those genera and tis recognition as a new genus, Schraderanthus. Phytologia 91(1):3-17 (April, 2009). KEY WORDS; Solanaceae, Athenaea, Chamaesaracha, Jaltomata, Leucophysalis, Physalis, Saracha, Witheringia, Mexico. A Mexican and Central American species that in most recent literature has been treated as Athenaea viscosa or Leucophysalis viscosa has been placed in seven different genera since its description as Saracha viscosa in 1832. It also was treated as a species of Physalis, requiring a new specific epithet because of the earlier name, P. viscosa L. Saracha viscosa was described from plants grown from seeds collected in Mexico but the species extends into Guatemala. Current data and generic concepts provide strong support for the recognition of this problematic species as a new monotypic genus. While the single species concerned is reasonably well-known, the following synopsis is provided. Schraderanthus Averett, Gen. nov. Herbae vel frutices usque ad 1-2.5 m altae; inflorescentiae fasciculatae axillares; calyx accrescens fructificans campanulatus, basi rotundatus, lobis 5 aequalibus, ad apiceum acutis, in fructu maturo campanulato- retrorsis, baccam rubro-aurantiaca, 10-15 mm diametro; semina ca 50- 75. Phytologia (April 2009) 91(1) 55 Erect herbs or soft-wooded shrubs 1.0-2.5 m tall, viscid, glandular-pubescent; inflorescences fasciculate with 6-8 (-10) flowers per axil, corolla 5-lobed with greenish maculations in the throat, ca 4 cm in diameter, rotate; anthers bluish (drying to a yellow-green), the filaments inserted at the base; flowering calyx accrescent, exceeding the length of the corolla, deeply lobed to ca % the length of the calyx, lobes acute; fruiting calyx broadly campanulate, deeply lobed, exceeding the berry but becoming reflexed at maturity, exposing the berry; berry bright red to orange-red, seeds ca 50-75, reniform, brown, ca 2 mm long, the testa rugose-reticulate. The most distinctive aspects of this novel genus are the 6-8 flowers in fascicles, as opposed to 1-2 per axil in some related genera and the bright red to orange-red berries surrounded by an accrescent calyx which initially loosely envelops the fruit and then opens into a broad campanulate to reflexed structure beneath the berry. These are features unknown in any of the genera to which the entity has been previously aligned. The nature of the calyx, in particular, is not always captured on dried specimens (including the type), and is not illustrated by Hunziker (2001), but is well illustrated in the Flora de Veracruz (Nee, 1986). The genus is named for Heinrich Schrader who first described the species concerned. Type species: Schraderanthus viscosus (Schrad.) Averett, Comb. nov. Basionym: Saracha viscosa Schrader, Index Seminum [G6ettingen] 5. 1832. TYPE: Cult., Hort. Géettingen, Schrader s.n. (MO, not seen;. phototype seen on TROPICOS). Synonymy: Physalis schraderiana Bernh., Linnaea 13: 361. 1839. Witheringia viscosa (Schrad.) Miers, Ann. Mag. Nat. Hist., ser. 2, 11(62): 92. 1853 Athenaea viscosa (Schrad.) Fernald, Proc. Amer. Acad. Arts 35: 567. 1900. 56 Phytologia (April 2009) 91(1) Jaltomata viscosa (Schrad.) D'Arcy & Til. Davis, Ann. Missouri Bot. Gard. 63: 363. 1976[1977]. Leucophysalis viscosa (Schrad.) Hunz., Kurtziana 21: 283. 1991. Chamaesaracha viscosa (Schrad.) Hunz., Lorentzia 8: 8. 1995. Complete descriptions of the species are provided by D’Arcy (1976) as Jaltomata viscosa, and as Leucophysalis viscosa (Hunziker, 2001). The species is also described as Athenaea viscosa in the Flora of Guatemala (Gentry and Standley, 1974) and in Spanish in the Flora de Veracruz (Nee, 1986). It is a very distinct species and unlikely to be confused with anything else except, perhaps, Brachistus nelsonii (Fernald) D’Arcy, J. Gentry & Averett and further description does not seem required. Schraderanthus viscosus occurs in the Mexican states of Chiapas, Oaxaca and Veracruz and extends into Guatemala. The location in Veracruz is from cited material (Nee, 1986); the later notes that the species as rare and little-known in Veracruz. ADDITIONAL SPECIMENS: GUATEMALA: Baja Verapaz: Union Barrios, west of km 154 on the Coban road, 12 Apr 1975. Lundell & Contreras 19171 (LL); MEXICO: Chiapas, Mpio. La Independencia, logging road from Las Margaritas to Campo Alegre, 2300 m, 18 Feb 1973, Breedlove 33631 (NY, TEX); Oaxaca, Distr. Mixe, 2 km N de San Miguel Metepec, 8 Apr 1984, Torres & Martinez 4967 (LL); Oaxaca, ca 6 km S of Totontepec, 18 Feb 1992, Panero & Campos 2761 (TEX); Distr. Ixtlan, Mpio. Santiago Comaltepec, Soyalapan, 100 m, 17°45’N, 96°30’W [imprecise], 16 May 1988, E. Lopez G. 120 (NY); Distr. Ixtlan, Sierra de Juarez, camino de Calpulalpan a Llano Verde, 12 km al NO de Calpulalpan, 2500 m, 29 May 1983, Lorence & Cedillo 4195 (NY); Sierra Mazateca, Mpio. Mazatlan Villa de Flores, San Pedro de los Encinos, 18°04’05.3”W, 96°52°41.9”"W, 2325 m, 23 Apr 2002, X. Munn-Estrada & Mendoza 2263 (NY); 50 km S de Valle Nacional, sobre la carretera a Oaxaca, 2250 m, 28 Jun 1975, Rzedowski 33382 (NY); Distr. Mixe, 5.2 km NE de la desviacion a Zacatepec, 2380 m, 23 Apr 1983, Torres & Cedillo 2680 (NY); Distr. Mixe, 7 km NE de la desviaciodn a Zacatepec, 2380 m, 23 Apr 1983, Torres & Cedillo 2697 (NY); Distr. Villa Alta, 11.7 km N de Maravillas a 39.7 km al N de Zoogocho, 2020 m, 15 May 1983, Torres et al. 2938 (NY); Distr. Mixe, 20 km N de Yacochi, camino a San Andrés Yaa, 2290 m, 8 Aug 1985, Torres et al. 7108 (NY). Veracruz, Mpio. Phytologia (April 2009) 91(1) 57 Atzalan, La Calavera, Carretera Altotonga-Tlapacoyan, Chdzarp & Dorantes 94 (ENCB). U.S.A. Massachusetts: “Hort. Cantab.” [Botanical Garden at Cambridge, Harvard University], 1849, 4. Gray s.n. (NY). Missouri: St. Louis Co., St. Louis, cult. by W. G. D’Arcy for the Second International Solanaceae Conference (“780425-2”), 6 Aug. 1982, M. Nee 25507 (NY); Switzerland. HBBasil [Botanical Garden at Basil], 29 Jun 1863, s.c. s.n. (NY).; Australia. South Australia: Southern Lofty Region, Tusmore (suburb of Adelaide), pot grown, provenance unknown, | May 1991, D. E. Symon s.n. (NY). DISCUSSION Saracha was once broadly construed to include a group of herbs widely distributed throughout Mexico, Central and South America that are now generally referred to Jaltomata (Gentry, 1973). Saracha is now recognized as a small genus of two species occurring at high elevations in South America with broadly or narrowly campanulate corollas and non or minimally accrescent calyces; it is part of a complex of several woody, mostly Andean genera, including Iochroma, Dunalia and Acnistus (Smith & Baum, 2006). D’Arcy and Davis (D’Arcy, 1976), following Gentry’s restoration of the genus Jaltomata and recognizing the species as separate from Saracha, transferred the species to the former as Jaltomata viscosa. He acknowledged, however, that its placement within Jaltomata was problematic. D’Arcy provided an excellent account of its history, with an expanded description of the taxon. Between the original description of Saracha viscosa and its transfer to Jaltomata, the species, besides to Physalis, had been assigned to Witheringia and Athenaea which, as constituted today, differ in a number of floral features and are clearly separate. Subsequent to the work of D’Arcy and Davis, Mione (pers. comm.) grew and became familiar with the species in question. Mione noted that the taxon has red fruit, which are absent in all Mexican Jaltomata; because of this, and characters of the fruiting calyx, he rejected placement of the species in Ja/tomata. Molecular studies (Mione et al., 1994) provide strong additional support for exclusion of 58 Phytologia (April 2009) 91(1) S. viscosus from that genus. Jaltomata possesses umbellate inflorescences which are not present in Schraderanthus, and the accrescent calyx is spreading in fruit. Hunziker (1991) transferred Jaltomata_ viscosa _ to Leucophysalis and subsequently (1995) to Chamaesaracha. In his Genera Solanacerum (2001), Hunziker returns the species to Leucophysalis, where he notes that the systematic position of the species has been in dispute and assigned to six different genera, mostly without explanation, which, it seems, would include his own transfers. Hunziker also notes the peculiar disjunctions within Leucophysalis. Leucophysalis grandiflora, the generiotype, has the northernmost distribution of any North American Solanaceae, while L. viscosa is neotropical. The characters that distinguish Schraderanthus from Jaltomata also distinguish it from Leucophysalis and Chamaesaracha. Excepting two doubtful species of sect. Capsicophysalis of the latter, red fruit are absent in Chamaesaracha and Leucophysalis. The flowers, with the broken green maculations at the base of the petals and distinctly lobed margins, also differ. Further, the wide disjunction between L. grandiflora and Schraderanthus and suggests a different origin. Whitson and Manos (2005) conducted a two-gene analysis from selected physaloid species that showed a close relationship between Leucophysalis grandiflora and L. nana, and a distant relationship between both species and Schraderanthus viscosus. A phylogenetic analysis of morphological characters by Axelius (1996) provided some evidence for a closer relationship between L. grandiflora and S. viscosus, but additional analyses of fruit and calyx characters of the fruit would be useful. Unfortunately, there are few phylogenetic analyses that include S. viscosus and the two species of Leucophysalis. In short, Chamaesaracha, including the type, C. coronopus, is Clearly a distinct assemblage. Gentry (1974) treated S. viscosus as Athenaea viscosa and noted that it to be morphologically similar to a group of Physalis species that have a 5-lobed corolla and mostly several flowers from Phytologia (April 2009) 91(1) 59 individual axils, but lacking the inflated fruiting calyx. A similar observation was made by Nee (1986) and Mione et al. (1994). Schraderanthus may be most closely related to Brachistus, Darcyanthus, and one or two species Hunziker (2001) have been treated as Chamaesaracha sect. Capsicophysalis,; all have red or orange-red fruit and share other characters (Table 1). However, lacking any clear evidence that any are congeneric, it seems most appropriate to recognize Schraderanthus as and distinct genus. Table | summarizes the salient characters of Schraderanthus and closely related genera. ACKNOWLEDGEMENTS I am grateful to Drs. Robert Hattaway, Thomas Mione, and Michael Nee for critical reviews of the manuscript. Dr. Mione also shared photographs and alcohol-preserved material of Schraderanthus which were especially helpful in understanding the floral and fruiting characters. Specimens were borrowed from the University of Texas Plant Resources Center (LL, TEX). Dr. Nee provided localities and other label data for the specimens at New York Botanical Garden (NY). Phytologia (April 2009) 91(1) 60 usai6 ‘Aujaq ou} uaa ‘Auiaq 3u} usaJ6 ‘Aaq Buisojous Ajjemed BHulsojous Jo Ajjemed pou ‘Ayunyeuw au} Bulsojoue pue 0} pue Burssaidde pue Bulssaidde ye Auiaq ay} Japun pessaidde ‘jusosas00V/ Ajasojo ‘jusoSaID0V Ajasojo ‘jJuasoses00y + = Bulxayjas ‘Juadsas00y xAjeo Buljini4 Ausaq Aysaly ‘pay Auiaq Ap ‘uaaig Assaq Aysal ‘uaaig Ausaq Aysal ‘pay wn pepyed-c peped jou ‘peqo|-g aijua Ajweou suibsew payed 0} paqo|-s ‘gyejnuedwed Ayyybijs ‘ayeyoy ‘paqo] aj}}!| ‘aJe}ON += ‘Paxayas pue 9}ejOY B||O10D sjixe siixe sjixe sjixe woud ‘ajejnoiose WO} SIOMO|} Z JO | WO} SIBMO|J Z JO | WwOd) ‘ajejnoiose4 |sdUadSaJ0}U| quay quay jejuudsed/jenuue $90} ||eWS40 jeluugied Bulpeaids = jejuuased Hulpeoids ‘snosoeqiay sqniys }oeJ9 Jo Bulpusosy JO jenuue ‘yoo19 Jo Apoom ‘jas —— . Accessed August 2008. Wunderlin, R.P. and B.F. Hansen. 2008. Atlas of Florida Vascular Plants [S.M. Landry and K.N. Campbell (application development), Florida Center for Community Design and Research.] Institute for Systematic Botany, Univ. of South Florida, Tampa. . Accessed August 2008. Phytologia (April 2009) 91(1) 69 TWO ADDITIONS TO THE VASCULAR FLORA OF TEXAS Jason R. Singhurst Wildlife Diversity Program Texas Parks and Wildlife Department, Austin, Texas 78704 U.S.A. jason.singhurst@tpwd.state.tx David J. Rosen S. M. Tracy Herbarium Department of Rangeland Ecology & Management Texas A&M University College Station, Texas 77843-2126, U.S.A. Walter C. Holmes Department of Biology Baylor University Waco, Texas 76798-7388 U.S.A. Walter_holmes@baylor.edu ABSTRACT Euthamia caroliniana is reported as new to Texas, while Rhynchospora chapmanii is reported as new to Texas and the West Gulf Coastal Plain. Phytologia 91(1):69-72 (April, 2009). KEY WORDS: Asteraceae, Cyperaceae, Euthamia, Rhynchospora, Louisiana, Texas, West Gulf Coastal Plain. The following species are reported as new to Texas. Euthamia caroliniana (L.) Greene ex Porter & Britton (Asteraceae). Euthamia caroliniana (Haines 2006) is distributed from Nova Scotia and Maine, south to Florida, and west to Michigan, Illinois, and Louisiana. Until now, it has not been reported in Texas (Correll & Johnston 1970, Turner et al. 2003, Haines 2006). The species was 70 Phytologia (April 2009) 91(1) collected at Candy Abshier Wildlife Management Area adjacent to Smith Point. This location is approximately 180 km west of the nearest known occurrence of £. caroliniana in Jefferson Davis Parish, Louisiana, where it is treated under the synonym Euthamia tenuifolia (Pursh.) Nuttall (Thomas and Allen 1996). Euthamia_ caroliniana occurred infrequently in a wet sandy coastal prairie and adjoining salt pan (slicks) in small patches that typically consisted of 5--10 plants. Several larger patches with 50-- 100 plants were also present. The site was dominated by Bigelowia nuttallii, Borrichia frutescens, Boltonia_ diffusa, Dichanthelium acuminatum, Eupatorium glaucescens, Fimbristylis castanea, Fuirena breviseta, Hypericum drummondii, Iva angustifolia, Liatris acidota, Lythrum alatum, Morella cerifera, Rhynchospora plumosa, Scleria georgiana, Schizachyrium tenerum, Solidago tortifolia, and Xyris stricta. Voucher specimen: TEXAS. Chambers Co.: Candy Abshier Wildlife Management Area, Smith Point at the southwestern tip of FM 562 at Galveston Bay, 4 Nov 2007, Singhurst 15467 (BAYLU). Euthamia is a small genus of five species, closely allied to, but distinct from Solidago (Haines 2006). Other species of the genus known to occur in the state include E. gymnospermoides Greene and E. leptocephala (Torrey & A. Gray) Greene (Turner et al. 2003, Haines 2006). A third species, E. graminifolia (L.) Nuttall is cited as occurring in Texas by Turner et al. (2006), but is excluded from the state by Haines (2006). A key to species and further information is available in Haines (2006). Rhynchospora chapmanii M.A. Curtis (Cyperaceae) In Godfrey and Wooten (1979) and Kral (2000) Rhynchospora chapmanii is considered to be endemic to the United States where it occurs from North Carolina, south to Florida, and west to extreme eastern Louisiana. There are, however, unpublished reports in the TROPICOS Database of the species occurring in Belize and Nicaragua (MOBOT 2008). Until the present paper, R. chapmanii had only been Phytologia (April 2009) 91(1) 71 documented in the Southern Atlantic and Eastern Gulf Coastal Plains. The record cited below constitutes the first known report of the species not only in Texas, but in the West Gulf Coastal Plain as defined by Bailey et al. 1994. This distribution pattern is common to a number of other Eastern Coastal Plain species, such as Agrimonia_incisa, Platanthera chapmanii, and Xyris smalliana, all present as disjuncts in southeast Texas, but with the major part of their distributions being from southeast Louisiana and eastward. Rhynchospora chapmanii was collected in a shallow wetland in a sandy coastal prairie in Candy Abshier Wildlife Management Area adjacent to Smith Point. The site was dominated by Anthaenantia rufa, Panicum rigidulum, Rhynchospora spp., Fuirena sp., and Eleocharis sp. This location is about 450 km west of the nearest known occurrence of R. chapmanii in St. Tammany Parish, Louisiana (Thomas and Allen 1993). TEXAS. Chambers Co.: Smith Point, Candy Abshier Wildlife Management Area, Take IH 10 E to SH 61 at Hankamer, S on SH 61 for four miles to FM 562, turn onto FM 562 and continue S then W for 22 miles on FM 562 to Smith Point, 01 Nov 2007, Rosen 4660 (BRIT, TEX, VDB. ACKNOWLEDGEMENTS We are grateful to Billie (B.L.) Turner and Tom Wendt for their assistance at the University of Texas Herbarium (TEX). We would also like to thank Robert Kral for his confirmation of Rhynchospora chapmanii. LITERATURE CITED Bailey, R.G., P.E. Avers, T. King, and W.H. Mc Nab. 1994. Ecoregions and subregions of the United States (map). Washington, D.C., U.S. Department of Agriculture-Forest Service, scale 1:7,500,000. Correll, D. S. and M. C. Johnston. 1970. Manual of the vascular plants of Texas. Texas Research Foundation, Renner. 72 Phytologia (April 2009) 91(1) Haines, A. 2006. Euthamia. In: Flora of North America Editorial Committee, eds. Flora of North America 20. Oxford Univ. Press, New York, pp. 97—100. Godfrey, R.K. and J.W. Wooten. 1979. Aquatic and wetland plants of the southeastern United States. Monocotyledons. Univ. of Georgia Press, Athens. Kral, R. 2000. Rhynchospora. In: Flora of North America Editorial Committee, eds. 1 Flora of North America 23. Oxford Univ. Press, New York, pp. 200-239. Thomas, R, D. and C. M. Allen. 1993. Atlas of the vascular flora of Louisiana. Vol. 1. Ferns & Fem Allies, Conifers, & Monocotyledons, Louisiana Department of Wildlife & Fisheries, Baton Rouge. Thomas, R, D. and C. M. Allen. 1996. Atlas of the vascular flora of Louisiana. Vol. II: Dicotyledons. Acanthaceae - Euphorbiaceae. Louisiana Department of Wildlife & Fisheries, Baton Rouge. MOBOT 2008. The TROPICOS Database (hitp://mobot.mobot.org/W3T/Search/vast.html, 14 Feb 2008) Missouri Botanical Garden, St. Louis, Missouri. Turner, B. L., H. Nichols, G. Denny, and O. Doron. 2003. Atlas of the vascular plants of Texas. Vols. 1 and 2. Sida Bot. Misc. 24. Botanical Research Institute of Texas, Fort Worth. Phytologia (April 2009) 91(1) 73 ROMULEA ROSEA (IRIDACEAE): ADVENTIVE IN TEXAS Jason R. Singhurst Wildlife Diversity Program Texas Parks and Wildlife Department, Austin, Texas 78704 U.S.A. jason.singhurst@tpwd.state.tx Kay M. Fleming 809 E. Clinton Ave. Athens, Texas 75751 U.S.A. Ruth Loper 13456 CR 2235 Whitehouse, Texas 75791 U.S.A. Virginia Privett 14839 CR3520 Brownsboro, Texas 75756 U.S.A. and Walter C. Holmes Department of Biology Baylor University Waco, Texas 76798-7388 U.S.A. walter_holmes@baylor.edu ABSTRACT Romulea rosea is reported as adventive in Texas. Phytologia 91(1):73-75 (April, 2009). KEY WORDS: Iridaceae, Romulea, Texas, invasive plant. Romulea is a genus of about 90 species that is distributed in Africa, southern Europe, and the Middle East (Goldblatt 2002). One 74 Phytologia (April 2009) 91(1) species, R. rosea, the sand crocus, a native of South Africa, is naturalized in Europe, the British Isles, Australia, New Zealand and in several coastal counties of California (The Nature Conservancy 2005). Based upon the specimens cited below, this species may now be reported as adventive in Texas. Romulea rosea (L.) Eckl. (Iridaceae). Voucher specimens: TEXAS. Henderson Co.: West side of County Road 3520, ca. 200 yards south of the junction of County Road 3520 and FM 2339, 24 Mar 2005, Fleming 1211 (BAYLU); West side of County Road 3520, ca. 200 yards south of the junction of County Road 3520 and FM 2339, 25 Mar 2005, Fleming 1212 (BAYLU). In 2005, the population consisted of approximately twenty individuals scattered over about one-half hectare under closed, undisturbed forest canopy dominated by Pinus taeda, Liquidambar styraciflua, Quercus spp., and Carya sp. In March 2008, the population was estimated to be “at least 100 plants over a larger area” (Fleming, pers. obs.). Five varieties of Romulea rosea are recognized by de Vos (1972). The specimens reported here correspond to variety R. australis (Ewart) M. P. de Vos. Goldblatt (2002) mentions that this variety has become a common weed of lawns, pathways, and roadsides in Australia, where it is a pest. He further mentions that, to date, this situation does not seem to be the case for the parts of California where the species occurs. While at present there is no evidence of the species becoming a pest in Texas, its invasive nature in Australia justifies that its presence in the state be closely monitored. Romulea rosea exhibits a pattern of spread that is similar to several other species reported as adventive in Texas. These species were initially reported as adventive to California (or the west coast), then later discovered in eastern Texas. However, the source of the plants adventive in Texas is unknown. Plants with this pattern of distribution include Bellardia trixago (L.) All. (Scrophulariaceae) (Lipscomb & Ajilvsgi 1982, Do et al. 1996), Centaurium muhlenbergii (Griseb.) Piper (Gentianaceae) (Holmes & Wivagg 1996), and Parentucellia viscosa (L.) Caruel (Scrophulariaceae) where it appears Phytologia (April 2009) 91(1) ff) to have been a “late” entry into the Manual of the Vascular Plants of Texas (Correll & Johnston 1970). Inclusion in the manual was based upon a 1969 collection (Correll 37239, TEX) of the species in Jasper Co. Tracing the application of the name in the state was complicated by the name not being included in the index of the manual. ACKNOWLEDGEMENTS We are grateful too Dr. Lynn Sherrod at University of Texas at Tyler for his assistance with determining the species of Romulea. LITERATURE CITED De Vos, M. P. 1972. The genus Romulea in South Africa. J. S. African Bot., suppl. 9. Correll, D. S. and M. C. Johnston. 1970. Manual of the Vascular Plants of Texas. Texas Research Foundation, Renner. Do, L. H., W. C. Holmes, and J. R. Singhurst. 1996. New County Records for Bellardia trixago (Scrophulariaceae) in Texas. Sida 17: 291-294. Goldblatt, P. 2002. Romulea. In: Flora of North America Editorial Committee, eds., Flora of North America North of Mexico 26. Magnoliophyta: Liliidae: Liliales and Orchidales. Oxford Univ. Press, New York and Oxford. p. 407 [illustration p. 406]. Holmes, W. C. and D. E. Wivagg. 1996. Identification and Distribution of Centaurium muhlenbergii and C. pulchellum (Gentianaceae) in Louisiana, Mississippi, and Texas. Phytologia 80: 23-29 Lipscomb, B. and G. Ajilvsgi. 1982. Bellardia trixago (L.) All. (Scrophulariaceae) adventive in Texas. Sida 9:370-374. The Nature Conservancy of Texas. 2005. Global Invasive Species Team. http://tncweeds.ucdavis.edu/alert/alrtromu.html, 20 March 2008. 76 Phytologia (April 2009) 91(1) TAXONOMY OF IVA ANGUSTIFOLIA AND I. ASPERIFOLIA (ASTERACEAE) B. L. Turner Plant Resources Center The University of Texas at Austin Austin, Texas 78712 billie@uts.cc.utexas.edu ABSTRACT Iva angustifolia and I. asperifolia are treated as belonging to a single species, /. asperifolia. The species is treated as having three varieties: var. angustifolia, widespread in the south-central U.S.A.; var. latior, largely confined to the beaches of southern Texas; and var. asperifolia, confined to the coastlines of central Mexico, with an introduced outlier in Wakulla Co., Florida. Keys to the taxa are provided, along with maps showing their distribution. Phytologia 91(1):76-83 (April, 2009). KEY WORDS: Asteraceae, [va angustifolia, I. asperifolia, I. texensis, Texas, Mexico, Florida Strother (2006), in his treatment of /va for the Flora of North America, provided a taxonomic account of /. angustifolia Nutt. ex DC. In this he did not provide for infraspecific categories, nor did he discuss its relationship to its closest congener, /. asperifolia, a taxon from dune sands along the Gulf Coast of Mexico. He did place in synonymy under his broad interpretation of /. angustifolia the names I. a. var. latior Shinners and J. texensis R.C. Jackson, both typified by material from sandy or clay saline dunes of southernmost coastal Texas. Jackson (1960) proposed the name /va texensis (typified by Jackson 2505 from saline soils ca. 8 mi S of Falfurrias, Brooks Co., Texas). Oddly, other than the type, he listed numerous additional collections, these introduced with the statement that “A number of specimens from the coastal regions of Texas are tentatively assigned to Phytologia (April 2009) 91(1) 7 I. texensis. These plants have a woody stem, but in involucral length and plant height are not always in agreement with the type.” Shinners (1964) inexplicably described again Jackson’s /va texensis with his proposal of /. angustifolia var. latior, the latter also typified by specimens from Brooks Co., Texas. Indeed, Shinners did not mention /. fexensis in his account of the proposed variety, although he did cite Jackson’s revision of /va in which the name was first proposed. Strother’s account of J/va angustifolia for the U.S.A. is confounded by the earlier name, /. asperifolia Lessing, this proposed in 1830. Strother did comment upon the latter, noting in the introduction to his treatment, that “Records of /va asperifolia Lessing from Florida are evidently based on specimens that are treated as members of /. angustifolia.” A similar sentiment was expressed by Wunderlin (1998) who thought the name /. asperifolia, was “misapplied” to plants from Wakulla Co. Florida, that he took to be /. angustifolia. This, in spite of the fact that Jackson (1960) stated, “The specimens of /. asperifolia from Florida may represent an introduction [of 4. asperifolia] from Mexico.” Plants grown from seeds by Jackson in his greenhouse (the seeds provided by Dr. R. K. Godfrey from Wakulla Co.) reportedly “gave plants with procumbent terms (sic).” Jackson further noted that the Mexican plants, which he observed along the beaches of Veracruz, Mexico, were similarly decumbent perennials that root at the nodes. One must assume that Jackson knew what he was talking about, hence the comments of Clewell (1985), no doubt bowing to the judgment of Jackson to the effect that the Wakulla populations, which he took to be I. asperifolia, are “decumbent perennials rooting at the nodes;” Clewell further noted that the plants occur in tidal marshland and were “Probably introduced from Mexico.” I have also examined collections from the Florida populations concerned and they appear to be closer in habit to /. asperifolia than to /. angustifolia, as noted by Jackson. Miao, Turner and Mabry (1995) examined chloroplast DNA variation in all species of /va, except for the mostly Cuban, /. cheiranthifolia. Their study included all four species of sect. Linearibractea R.C. Jackson, which included /. angustifolia, I. asperifolia, I. texensis, and I. microcephala. Indeed, in their sequence divergence study the four species of sect. Linearibractea formed a tight 78 Phytologia (April 2009) 91(1) cluster, the most distinct being /. microcephala, which was found to be the most divergent member of the tetrad. The remaining three taxa were found to have few differences among them. In particular, the two herbaceous taxa, /. angustifolia and I. asperifolia, were found to differ by only one restriction site mutation. Unfortunately, DNA data for J. asperifolia was obtained from the Wakulla Co. population, which Jackson accepted as an introduction from the Mexican beaches. The taxonomic status of the latter population is controversial, as noted in the above. Considering the Pleistocene history of the southeastern U.S.A., it is possible that the Wakulla population is but an isolated relic of a once more widespread, highly variable, var. angustifolia, perhaps deserving of its own formal varietal name. Additional study of the /. asperifolia complex is clearly needed, especially from Tamaulipas where the two taxa presumably intergrade. With the above as an introduction, I hasten to add that I also (Turner et al. 2003) recognized Texas populations of the J. asperifolia complex as J. angustifolia, largely following the work of Jackson. Unlike Strother, I did recognize the more robust, seemingly perennial, var. latior of Shinners, with the assumption that it intergraded northwards along the shores of Texas into the typical var. angustifolia, as also noted by Jackson. I also believe that the var. /atior intergrades southwards along the Gulf beaches of Mexico into var. asperifolia. This suggested by a single intermediate, cited below. I provide herein a synopsis of the /. asperifolia complex as currently understood, along with a nomenclature that appears to fit the biology. IVA ASPERIFOLIA Lessing, Linnaea 5: 151. 1830. TYPE: MEXICO. VERACRUZ: Mpio. Veracruz, “In pascuis pr. Vera-Cruz.” Jul. Scheide 332 (holotype B; fragment and drawing, GH). Jackson (1960) provided a detailed description of the taxon, positioning it in his sect. Linearbractea. Phytologia (April 2009) 91(1) 79 Key to infraspecific taxa: 1. Plants annuals mostly 0.75 m high or less; south-central U.S.A. Ole as Pexts Ark: s Lai )ic. oi) aad id ne caWedenee tenses var. angustifolia 1. Plants perennial herbs to 1.5 m high; Gulf Coastal dunes, southernmost Texas, northern Mexico and panhandle Florida...... (2) 2. Stems decumbent, rooting at the nodes; leaves mostly oblong- lanceolate; involucres without markedly spreading, broad-based hairs; coastal Tamaulipas and Veracruz, Mexico...... var. asperifolia 2. Stems stiffly erect, only rarely rooting at the nodes; leaves mostly lanceolate; involucres with markedly broad-based spreading hairs, rarely not; northern coastal Tamaulipas, Mexico, and southern Texas WRB SCG Saatste devin cinder abs cctlin uote Ue tas cadebcneaeaeeandethos var. latior IVA ASPERIFOLIA var. ANGUSTIFOLIA (Nutt. ex DC.) B.L Turner, stat. & comb. nov. Fig. | Based upon /va angustifolia Nutt. ex DC., Prodr. 5: 529. 1836. TYPE: U.S.A. ARKANSAS: without locality, Nuttall s.n. (holotype: G-DC; isotype NY) Jackson (1960) provided an adequate account of this taxon, noting in his key the principal differences that distinguish it from var. latior, the latter being larger, mostly perennial, plants having somewhat larger involucres, the latter well endowed with spreading hairs. In my opinion, var. angustifolia grades into var. /atior in regions of near contact, as noted below, hence their treatment as but populational variants (or variety) of a widespread /va asperifolia. Varity angustifolia is highly variable and, other than habit, is best recognized by a suite of characters, most having to do with the involucre, as noted below and by Jackson (1960). IVA ASPERIFOLIA Lessing var. ASPERIFOLIA Fig. 1, 2 As noted in Figures 1 and 2 (based upon specimens at MEXU, TEX), this taxon is restricted to the Gulf Coastal area of northern Tamaulipas and Veracruz, Mexico, with an isolated population in 80 Phytologia (April 2009) 91(1) Wakulla Co., Florida. In Tamaulipas, Mexico, it seemingly grades into the var. /atior, to judge from the following collection: Mpio. Altamira, Dunas 856 (MEXU, TEX). The taxonomic status of the Florida populations of var. asperifolia is in doubt. According to label data on Godfrey 70060 (TEX), the Wakulla population occurs “On a large flat area into which spoils from dredging of the St. Marks River had been placed; very abundant; St. Marks.” The DNA voucher of Miao et al. (1995) was obtained “In limestone piles of spoil, St. Marks, just west of end of county Road 363 and north of St. Marks River.” Both of the aforementioned collections are seemingly perennial. The Godfrey collection has a capitulescence much resembling var. angustifolia; the Miao voucher (Garland 751, TEX) has an atypical capitulescence. Based on the DNA data from the latter voucher and the comments of Jackson (1960), it would appear that the Wakulla population is closest to var. asperifolia. Alternatively, it is remotely possible that the Wakulla populations represent a localized yet undescribed variety of J. asperifolia, as noted above. IVA ASPERIFOLIA var. LATIOR (Shinners) B.L. Turner, comb. nov. Fig.1 Based upon /va angustifolia var. latior Shinners, Sida 1: 378. 1964. TYPE: U.S.A. TEXAS: Brooks Co., “south of Falfurrias.” 15 Sep 1942, Lundell & Lundell 11947 (holotype SMU; isotypes LL!) Iva texensis R.C. Jackson, Univ. Kansas Sci. Bull. 41: 807. 1960. TYPE: U.S.A. TEXAS: Brooks Co., 8.1 mi S of Falfurrias along U.S. highway 281, 24 Aug 1957, Jackson 2505 (holotype KANU; isotype SMU) Jackson’s Iva texensis was published prior to Shinner’s var. latior; unfortunately nomenclatural priority is predicated upon rank; were the present taxon recognized as a species, its correct name would be 1. texensis! Variety /atior is a perennial herb to 1.5 m high, largely confined to the Gulf Coastal region of Texas, mostly occurring in sandy Phytologia (April 2009) 91(1) 81 or clay dunes along the beach front; numerous collections are in the herbarium at TEX (35 from along the coastal areas of Cameron, Kenedy and Nueces counties). The earliest collections of var. /atior were made by Robert Runyon in Cameron County between the years 1924-1945 (TEX, LL). He noted that the taxon was sufficiently well known as to have received the local common name “Pelocote.” And that the taxon occurs “only along sea shore as colonies on sand dunes;” and describes its habit as an “erect branched herb,” or “fruticose herb.” Nevertheless, it also occurs sporadically inland into the Tamaulipan Biotic Province of southern Texas, as indicated in Fig. 1. Occasional intergrades between var. latior and var. angustifolia in this area are discerned (e.g., Carr 22574, Refugio Co., TEX). The more strictly coastal populations of var. /atior pass into var. angustifolia near Galveston, Texas, the latter becoming strictly taprooted annual herbs mostly 0.5 m high or less, having smaller involucres with mostly shorter appressed hairs. As suggested by its dune-site proclivities in Kenedy and Cameron counties, it is almost certain that southwards var. /atior grades into var. asperifolia, as suggested by the intermediate specimen cited above. Even among the Texas coastal populations, some degree of past introgression is likely, to judge from the variation in leaf shape and pubescence noted among the Cameron county populations. ACKNOWLEDGEMENTS I am grateful to my colleagues, Guy Nesom and John Strother for reviewing the paper and providing helpful comments. Thanks also to MEXU for the loan of specimens, these critical in the construction of distribution maps for /va asperifolia var. asperifolia. LITERATURE CITED Clewell, A.F. 1985. Guide to the Vascular Plants of the Florida Panhandle. Florida State Univ. Press, Tallahassee Jackson, R.C. 1960. A revision of the genus /va L. Univ. Kansas Sci. Bull. 41: 793-875. 82 Phytologia (April 2009) 91(1) Miao, B., B.L. Turner and T.J. Mabry. 1995. Molecular phylogeny of Iva (Asteraceae, Heliantheae) based on chloroplast DNA restriction site variation. Pl. Syst. Evol. 195: 1-12. Strother, J.L. 2006. Iva, in Fl. N. Amer. (N. of Mexico.) 21: 25-28. Turer, B.L., H. Nichols, G. Denny and O. Doron. 2003. Atlas of the Vascular Plants of Texas 1: 1-648. Wunderlin, R.P. 1998. Guide to the Vascular Plants of Florida. Univ. Press, Florida asperifolia ® var. angustifolia © var. latior * var. asperifolia Fig.1. Distribution by county (and parish) of /va asperifolia in the U.S:A: Phytologia (April 2009) 91(1) 83 asperifolia o var. asperifolia © intermediate e var. latior Fig. 2. Distribution of /va asperifolia in Mexico. 84 Phytologia (April 2009) 91(1) A NEW SPECIES OF TRIXTIS (ASTERACEAE: MUTISIEAE) FROM JALISCO, MEXICO B. L. Turner Plant Resources Center The University of Texas at Austin Austin, Texas 78712 billie@uts.cc.utexas.edu Phytologia 91(1):84-87, (April, 2009) Preparation of a treatment for the Mutisieae of Mexico has occasioned description of the following novelty: TRIXIS JALISCANA B.L. Turner, sp. nov. Fig. | Trixi haenkei Schultz-Bip. similes sed foliis juvenilibus subtus dense villosis (vs moderate vel sparsim villosis), setis pappi fulvis (vs albis), et capitulescentia congestae (vs expansae). TYPE: MEXICO. JALISCO: Mpio. Mascota, 1 km E of Mascota, “dry, shrubby slopes,” 1230 m, 1 Mar 1970, W.R. Anderson & C. Anderson 5905 (Holotype: LL). Shrub 1-3 m high, “branching from the base and from lax branches.” Stems wingless, densely soft-pubescent. Leaves lanceolate, 5-15 cm long, 1-4 cm wide; petioles 0-10 mm long. Capitulescence a terminal, tightly congested, corymbose panicle, the ultimate peduncles 0-1 mm long. Accessory bracts 2-4, lanceolate, mostly half as long or less than the involucre. Involucral bracts 8, ca 12 mm long, their apices abruptly acute. Receptacle ca 2 mm wide, pubescent with hairs up to 2 mm long. Florets 10-20 per head; corollas yellow, the upper lip 3-4 mm long, the tube 8-10 mm long. Achenes pubescent throughout, 6-9 mm long; pappus of numerous tawny bristles 8-10 mm long. ADDITIONAL SPECIMENS EXAMINED: MEXICO. JALISCO: Mpio. Chapala, “Barranca proxima a Ajijic,” 1650 m, 12 Nov 1978, Luna 9370 (TEX). Mpio. Jocotepec, “Ladera sur al norte de la barranca del Huazoyo.” 7 Mar 1993 Machuca N. 6923 (TEX). Mpio. Mixtlan, between Ameca and Atenguillo, 5100-5500 ft., 4 Mar 1987, Phytologia (April 2009) 91(1) 85 Daniel & Bartholomew 4830 (TEX). Mpio. San Sabastian, N of Mascota on road to San Sebastian, 1450-1730 m, 1-3 Mar 1970, Anderson & Anderson 5951 (TEX). In her seminal treatment of Mexican 7rixis, Anderson (1972) clearly called to the fore the present novelty. She noted that the type of T. haenkei was probably collected by Seemann “in the mountains along the Sinaloa-Durango border, east of Mazatlan.” At the time of her study Anderson knew of only two collections from the area of the type locality concerned, the type and Anderson & Anderson 6177. She further noted that the latter collection differed from “the type as well as from the Jalisco collections” in having a white pappus, and inflorescence branches which bear small corymbs that are more than twice as long as those in specimens from Jalisco. I have examined the Anderson & Anderson collection and one additional collection from the area of the type locality: Rito Vega 3624 (TEX), this from Sinaloa between km 240 and 256 along the Ojo de Agua to la Guayamera highway. Both of these collections are very similar, and the characters called to the fore by Anderson hold for both. I have not examined the holotype itself, which is reportedly at Kew, nor did Anderson. She did, however, examine a photo of the holotype, as have I. From the photo | found it difficult to judge the characters called to the fore by Anderson. I did, however, borrow what I take to be an isotype from GH and found this to be closely akin to the other two collections from the Sinaloa- Durango border. In short, all material from along the Sinaloa-Durango border is clearly the same and is not to be confused with the presently described 7. jaliscana, T. haenkei occurring in tropical deciduous forests (600-800 m) “just below the oak zone,” and T. jaliscana occurring in pine-oak forests at much higher elevations (1200-2000 m). Further, among the Jalisco collections I detected no intermediates between the two regional populations concerned, which leads me to believe that the taxa are sufficiently distinct, both morphologically and ecologically, for specific recognition (as opposed to varietal). 1. Young leaves moderately to sparsely densely villous beneath; pappus bristles white; capitulescence open; deciduous forests below the oak LEE EEG RE NE Rs a T. haenkei 1. Young leaves densely villous beneath; pappus bristles tawny; capitulescence tightly congested; pine-oak forests..........T. jaliscana 86 TAT OOO E Fig. 1. Holotype of Trixis jaliscana. Phytologia (April 2009) 91(1) LUNDELL | | HERBARIUM | Ei Sins Thee Vienionrsity: oh Teaane AUSTIN, TEMAS pom erneraattmrnrtenericg : ; Cites 4h The Univonvety tl Tena tincnaetiaen ORNS Trine jabincams WL. Turner Gut, BL. Tiemer Bet PLANTS OF JALISCO, MEKICO atl esamentiontarnare iki ken CT AAR NBD, TRG ame KERR ROSIE Ramet BNE HA ie Um Ie: Triste beenket Uchaeliip, in Gemma Ory, ahrubhy wlopes atone 1 kw & or Mansotay eler. 129 9, Amadt ahah to 2 me tall, berenond ng trom the bane and fem tke Ya teraeches ¢ athitatant, © ANCE aE & Renee MeCN we, 9G 4 neces Phytologia (April 2009) 91 (1) 87 ACKNOWLEDGEMENTS I am indebted to Guy Nesom for the Latin diagnosis, and for improving the manuscript itself. I am also grateful to GH for loan of type material. LITERATURE CITED Anderson, C.E. 1972. A monograph of the Mexican and Central American species of Trixis (Compositae). Mem. New York Bot. Gard. 22: 1-68. McVaugh, R. 1984. Trixis, in Fl. Novo-Galiciana 12: 953-963. TRIXIS o haenkei @ jaliscana Fig. 2. Distribution of Trixis haenkei (circles) and T. jaliscana (dots). 88 Phytologia (April 2009) 91(1) CARMINATIA PAPAGAYANA (ASTERACEAE: EUPATORIEAE), A NEW SPECIES FROM WESTERN GUERRERO, MEXICO B. L. Turner Plant Resources Center The University of Texas at Austin Austin, Texas 78712 billie@uts.cc.utexas.edu Phytologia 91(1):88-91 (April, 2009) Routine identification of Mexican Asteraceae has revealed the following novelty: CARMINATIA PAPAGAYANA B.L. Turner, sp. nov. Fig. | Carminatia recondita McVaugh similis sed differt capitulescentia stricta laxa corymboidei capitulis in pedunculis ultimis 1-4 cm longis portatis (vs. capitulis congestis in pedunculis 1-3 mm longis). Annual herbs 20-70 cm high. Stems (lower) ca 3 mm across, pubescent with crinkly hairs. Leaves (lower) 4-5 cm long, 2-3 cm wide; petioles 2-3 cm long, glabrous or nearly so; blades broadly ovate to deltoid, glabrous above and below , the margins somewhat or weakly dentate. Capitulescence a terminal array of stiffly erect peduncles, the ultimate peduncles 1-4 cm long, pubescent like the stems. Heads cylindric, containing ca 15 florets, ca 14 mm high, 4-5 mm across. Involucres ca 12 mm high, glabrous; outer bracts 4-8, 1-6 mm long. Receptacle ca 1 mm across, glabrous. Corollas white, glabrous, cylindrical, 7-8 mm long, ca 0.8 mm wide. Achenes 4-5 ribbed, minutely pubescent with very short, broad-based hairs; pappus of ca 15 white, plumose, bristles ca 7 mm long; chromosome number, 2n = 20. TYPE: MEXICO. GUERRERO: Mpio. Tierro Colorado, "Acapulco. Autopista del Sol Mexico-Acapulco, zona rocosa a unos metros del puente sobre el Rio Papagayo. ..Selva baja caducifolia. Phytologia (April 2009) 91(1) 89 Occasional 280 m," 17 08 02.9 N 99 33 24.2 W, 9 Oct 1995, Jose L. Panero 6193 [with C. C. Clevinger] (Holotype: TEX). As indicated in Fig. 1, the present novelty differs from all previously described species in having its heads arranged in stiffly erect peduncles 1-3 cm long. It also appears to be restricted to deciduous forests along the lower slopes of the Sierra Madre of western Guerrero. Yet other species of the genus in Guerrero occur at higher, more eastern locales, as noted in Fig. 2, the latter based upon specimens at LL, TEX and those mapped in Turner (1997). Strother and Panero (2001) report a chromosome number of 2n = 20 for the type (identified as C. recondita McVaugh), this consistent with previous reports for the genus. The taxon is named for the Papagayo River, from whence the type and only known collection. ACKNOWLEDGEMENTS I am grateful to Guy Nesom for the Latin diagnosis and helpful comments on the manuscript itself. LITERATURE CITED Strother, J.L. and J.L. Panero. 2001. Chromosome studies: Mexican Compositae. Amer. J. Bot. 88: 499-502. Turner, B.L. 1997. Carminatia, in Phytologia Memoirs 11, The Comps of Mexico 1: 84-86. 90 Phytologia (April 2009) 91(1) UNIVERSITY OF EK AS HERG ANION ees : iin oi BIBLE ANE! palin BAH Wane BENS Re me ML, oa Va j SUS nnn scene ME PLANTAS DE MEXICO mnt Carminatia revomsdhite MeVaugety 4 +n ek tnid Setiradcns. Chr demew a venchewe 2 2M x has pra 4 Ard panewie witha 93 Bane Yonaneen i his vee Stather + Panere | i007} : ee aa vi tore:ne fe satowcstlins {aaron 28 W O949 hai 8. Parnes aes PRAT TIS Weta C. Clewin PRS ES GCS MR HS ONTO RAST C8 TRAN ERR AReOM ert Fig. 1. Carminatia papagayana, holotype. Phytologia (April 2009) 91(1) CARMINATIA A alvarezii * papagayana * recondita © tenuiflora Map 1 Fig. 2. Distribution of Carminatia spp. in Mexico. 9] 92 Phytologia (April 2009) 91(1) DELWIENSIA, A NEW GENUS OF ASTERACEAE W. A. Weber, F.L.S. & R. C. Wittmann University of Colorado Museum of Natural History UCB 265, Boulder, Colorado 80309 bill.weber@colorado.edu KEY WORDS: Asteraceae, Artemisia pattersonii, Delwiensia pattersonii. Phytologia 91(1):92-94 (April, 2009). The genus Delwiensia is proposed to accommodate Artemisia pattersonii A. Gray on cytological and morphological grounds. Delwiensia W. A. Weber & R. C. Wittmann, gen. nov., Asteraceae. Type, Artemisia pattersonii A. Gray, Syn. Fl. N. Amer. ed. 2, 1(2):453.1886. The genus name honors Delbert Wiens, contemporary American plant taxonomist, specialist in the Viscaceae, plant reproduction concerning embryonic abortion, and pollination by small mammals. Herba aspectu speciei Artemisiae in subgenere Absinthio, numero chromosomatico n = 7, habitu inflorescentiae determinato. Herb with the appearance of a species of Artemisia in the subgenus Absinthium, with the chromosome number n = 7, and with the inflorescence determinate. Delwiensia pattersonii (A. Gray) W. A. Weber & R. C. Wittmann, comb. nov. Basionym: Artemisia pattersonii A. Gray. Delwiensia is a monotypic genus endemic to the alpine tundra of Colorado, Wyoming, and New Mexico. It is superficially similar to Artemisia scopulorum but is amply distinct morphologically, its most obvious characters being the smaller number of heads in determinate rather than indeterminate arrays. Wiens and Richter (1966) point out that the root systems of the two species differ, A. pattersonii reproducing vegetatively from branched rhizomes and A. scopulorum having an unbranched caudex. Furthermore, A. pattersonii is unique in Artemisia in having a chromosome number of n = 7. The known base Phytologia (April 2009) 91(1) 93 numbers in Artemisia are x = 8 and predominantly x = 9. The details of the karyotype did not support the notion that the two species are related. In Sphaeromeria (see discussion below) the chromosome number is x = 9. It is of course possible that De/wiensia pattersonii might be found in the Asiatic flora and thus would be considered a Tertiary relic (Weber 2002), but cytological and morphological information need to be obtained for the numerous Asiatic species of Artemisia-like genera and species. Rydberg (1929) had the following pungent remarks about the inflorescences (from Scylla or Charybdis). This lumping, advocated by Hall and Clements, has also. been practiced by them. They include Sphaeromeria Nutt., Vesicarpa and Chamartemisia in Tanacetum. The only characters that Hall and Clements have left to separate Tanacetum and Artemisia are “Inflorescence cymose, the cyme occasionally reduced to a single head” in Tanacetum and “Inflorescence racemose-paniculate” in Artemisia. The inflorescence, strictly speaking, is neither cymose nor racemose-paniculate in either, for both have the flowers in heads. In my own treatise of the group I have used the word corymbiform instead of cymose, which I think is much better. Whether the heads are arranged cymosely or racemosely is hard to tell. If cymosely, the terminal and central head should be best and first developed. In such a case Artemisia pattersonii should be included in Tanacetum. The heads of that species have exactly the same arrangement as in Chamartemisia compacta or Sphaeromeria simplex, that is, usually one or two heads, the terminal one the larger. If the heads are congregated into a spherical cluster as in Artemisia glomerata and A. globularia and Sphaeromeria capitata (according to Hall and Clements a Tanacetum), it would be hard for anyone to tell whether the heads are arranged cymosely or racemosely. In Vesicarpa potentilloides (also a Tanacetum) [Sphaeromeria potentilloides (A. Gray) Heller] I cannot tell if the inflorescence is racemose 94 Phytologia (April 2009) 91(1) or cymose and I have dabbled in taxonomy for 50 years. Shultz (2006) distinguishes A. scopulorum (heads in spiciform arrays, and corolla lobes hairy) from A. pattersonii (heads being borne singly or 2-5, spreading to nodding, pedunculate, in paniculiform or racemiform arrays, and corollas mostly glabrous). The inflorescence can be better described as being determinate versus indeterminate. Rydberg’s explanation is to this point. ACKNOWLEDGEMENTS We thank Patricia Eckel for refining our Latin description. LITERATURE CITED Rydberg, Per Axel. 1929. Scylla or Charybdis. Proc. International Congress Plant Sciences 2:1539-1551. Shultz, Leila M. 2006. Artemisia, in Flora of North America, Vol. 19. Flora of North America Editorial Committee. Oxford Univ. Press. Weber, William A. 2003. The Middle Asiatic element in the Southern Rocky Mountain flora of the western United States: A critical biogeographical review. J. Biogeography 30:649-685. Wiens, Delbert, & Judith A. Richter, 1966. Artemisia pattersonii: a 14- chromosome species of Alpine Sage. Amer J. Bot. 55:981— 986. Phytologia (April 2009) 91 (1) 95 ZAMIA FLORIDANA (ZAMIACEAE), THE CORRECT NAME OF THE FLORIDA CYCAD Daniel B. Ward Department of Botany, University of Florida Gainesville, Florida 32611, U.S.A. ABSTRACT The view is accepted that the Zamia native to the West Indies consists of several species, one of which is native also to Florida. The earliest available binomial for the Florida taxon is Zamia floridana A. DeCandolle (1868). An earlier binomial, Zamia integrifolia Linnaeus filius in Aiton (1789), by citation in synonymy of the prior Zamia pumila Linnaeus (1763), was superfluous when published and is thus illegitimate. Phytologia 91(1):95-104 (April, 2009). KEY WORDS: Zamia floridana, Zamiaceae, nomenclature. INTRODUCTION The West Indian complex of cycads in the genus Zamia (Zamiaceae) has been treated by Eckenwalder (1980) as composed of a single species with populations that vary in leaflet width and vein number but are not appropriately divided into more than a single species, Z. pumila L. (1763). This interpretation has been accepted by some authors (e.g. Wunderlin, 1998; Wunderlin & Hansen, 2000), and Z. pumila is frequently used in Florida horticulture. A subsequent review of the West Indian cycads by Stevenson (1987a; 1987b; 1991), which incorporated leaflet shape and denticulation and cone shape and color, was able to distinguish 6 species within that area, one of which (his Zamia integrifolia) ranges to Florida. Landry (1993) in the influential Flora North America followed Stevenson in recognizing the Florida plant as specifically distinct from 96 Phytologia (April 2009) 91(1) the all-inclusive Z. pumila of Eckenwalder; Landry too employed Z. integrifolia. The present author has long been of the opinion that Zamia integrifolia L. f. in Ait. (1789) was nomenclaturally superfluous when published, in that Linnaeus filius (in Aiton) had erred (by modern rules) by citing in his synonymy a pre-existing name, Zamia pumila L. (1763). In this belief, the present author (1968; 1979; Burch et al., 1988) has consistently used a later available name, Z. floridana A. DC. (1868). In need of an infraspecific name for a non-typical Florida population, he made the combination, Z. floridana var. umbrosa (Small) D. B. Ward (2001). The authors of a recent, highly acclaimed systematics text (Judd et al., 1999: 151) have accepted this judgment, also using Z. floridana. DISCUSSION A circumstance has now arisen that compels presentation of a full defense of Zamia floridana. The recent proposal by a colleague to use this name in a floristic survey has by editorial review triggered an intense reconsideration of its nomenclatural underpinnings. To satisfy all parties that this name is correct calls for a full discussion of the background facts and provisions of the International Code of Botanical Nomenclature (McNeill et al., 2006) that justify this conclusion. The facts of publication seem not in dispute. In 1763 Linnaeus published the name Zamia pumila. He accompanied the name with a 7-word Latin phrase: Spadix more fructus Cupressi divisus in floscules ("Infructescence [=cone] larger than [that of] Cupressus, divided into florets [=?microsporophylls]"). He stated its source: Habitat in America meridionali. He then listed four earlier authors (P. Miller, J. Commelin ("Commelijn"), L. Plukenet, and C. J. Trew), with the phrase-names used by each. Two centuries later the illustration of one of these cited authors, that of Commelin (1697), was designated by Eckenwalder (1980: 715) as the lectotype for Z. pumila. Phytologia (April 2009) 91(1) 97 Linnaeus filius' treatment of Zamia was wholly independent of that of his father. He is known to have worked in London with William Aiton (Stafleu, 1971) and to have assisted in the writing of Aiton's Hortus Kewensis (1789); each of the five descriptions of Zamia in that publication was credited to him. In this endeavor he had access to living plants (he noted Z. integrifolia to have been introduced from "East Florida" by John Ellis in 1768). His description of Z. integrifolia (foliolis subintegerrimis obtusiusculis muticis rectis nitidis, stipite inermi) was original and appropriate to Florida plants. He cited only one reference, the Z. pumila of his father, but for this he stated "exclusis synonymis." This two-word Latin phrase is the genesis of all later nomenclatural uncertainty regarding the Florida Zamia. The third name involved here is Zamia floridana A. DC. (1868). Its author reported it from "E. Florida" and its label data (Eckenwalder, 1980) further narrowed its source to "Fort Brooke," a Seminole War army encampment at the head of Tampa Bay on the west coast of peninsular Florida, as collected by "Hulse." (In the 18th and early 19th century, all of peninsular Florida was in the political district of "East Florida." Gilbert White Hulse, a correspondent of John Torrey in New York, was a physician known to have been stationed at Fort Brooke.) The legitimacy of Z. floridana has not been challenged; but it rises from synonymy only in the event of the illegitimacy of the prior Z. integrifolia. On the surface, Linnaeus filius' inclusion of an available name would appear to trigger citation of I.C.B.N., Art. 52.1, which states that if an old name cited in synonymy could have been used for the new taxon, the new name is superfluous and illegitimate. Were Zamia integrifolia illegitimate, the later Z. floridana would succeed. Correspondents (largely via e-mail), however, have raised a number of questions and arguments directed toward invalidation of the apparent 1.C.B.N. citation, in part by invocation of the related Art. 52.2, thus retaining Z. integrifolia as legitimate. These communications have been widely circulated within the taxonomic/nomenclatural community, and have come to form a "gray literature" suggesting the validity of 98 Phytologia (April 2009) 91(1) Zamia integrifolia is established. It is these questions and arguments that must here be detailed and refuted. Since the written (e-mail) statements of correspondents had not been polished for publication, and at times perhaps exhibit whimsy over precision, their specific authorship is withheld. Unattributed statements, of course, are atypical within scientific discourse. To mitigate the conflicting goals of confidentiality and verifiability, a full copy of communications has been provided the editor. Where possible, statements of correspondents are cited exactly, as indicated by enclosure within quote marks. The following eight arguments well summarize the range of views expressed by the correspondents. The associated responses attempt to relate these remarks with relevant provisions of the I.C.B.N. Argument #1. That Linnaeus filius "meant to say just the opposite, 1.e. ‘excluding Z. pumila L. except the synonyms.' Perhaps a Latin scholar could refute my supposition that 'exclusis synonymis' can be read as 'including only the synonyms."" Response. No deep schooling in Latin is needed to know that "exclusis" cannot be interpreted to mean "including only." The logic and motive of Linnaeus filius in excluding his father's references is apparent, in that some addressed quite different plants (one became Zamia furfuracea L. f. in Ait.) and in any event were surely inferior to the far greater wealth of materials (living and dried) available to him in London. Argument #2. That Eckenwalder's designation of the 1697 Commelin illustration as the lectotype of Zamia pumila, which Linnaeus filius had excluded from his treatment of Z. integrifolia, removes the critical element -- the type -- from the citation, thereby expunging any prior superfluity. Response. Eckenwalder's lectotypification is irrelevant in determination of superfluity. If Zamia integrifolia were illegitimate prior to Eckenwalder's designation, barring certain circumstances it must remain so in perpetuity. Article 6.4 provides that: "A name which Phytologia (April 2009) 91(1) 99 according to this Code was illegitimate when published cannot become legitimate later" (unless conserved or sanctioned). Argument #3. That Linnaeus filius had in effect created a nomen nudum by exclusion of his father's cited references. [First correspondent]: "When L. f. excluded all the synonyms of Z. pumila, he automatically excluded all the type elements that would otherwise cause the superfluity.". [Second correspondent]: "Since Aiton clearly excludes the type of Zamia pumila of Linnaeus by excluding all the synonymy of Z. pumila, he created a new and valid name, Zamia integrifolia Aiton." Response. This argument is in reference to Art. 52.2, the companion of Art. 52.1, where conditions are set under which citation of an old name in synonymy will cause the new name to become superfluous. Citation of the name itself is specifically stated to be sufficient to cause superfluity, "unless the type is at the same time excluded either explicitly or by implication." But with Zamia pumila no type existed at publication. Nor was the legitimacy of that name impaired by its absence. Even without the synonyms, the citation of Z. pumila still encompassed a full circumscription: the name, the source, and the seven-word diagnosis. At that time, 1789, no "type element" was essential for valid publication; designation of a type was not required until 1958 (Art. 37.1). An abundance of early names, some by Linnaeus, many by Rafinesque and Thomas Walter among others, are based solely (if insecurely) on a name and its diagnosis. Though there is a seductive logic in equating the references cited by Linnaeus -- from which a later type-equivalent may be chosen -- with a type itself, the provisions of Art. 52.2, read critically, do not support the argument. Argument #4. That a party other than the original author has the power to cause the type of a name used in synonymy to be excluded. That is, the requirements of the I.C.B.N. for a superfluity- 100 Phytologia (April 2009) 91(1) causing synonym to be intentionally included are not met if a second party can cause the exclusion. Response. This argument, also in reference to the companion Art. 52.2, though not expressly stated by any correspondent, is implicit if the act of lectotypification can assign the type to a component of the original material of Zamia pumila that Linnaeus filius had excluded. Yet Art. 52.2 clearly indicates otherwise. Though the language is passive -- "unless the type is at the same time excluded either explicitly or by implication" -- there is no provision for parties other than the original author to cause such exclusion. Nor, of course, can a later party act "at the same time" as the original author. Argument #5. [First correspondent]: That "Z. integrifolia is not illegitimate because it did not include ALL the elements that might become the lectotype. In short, it was not superfluous at birth." [Second correspondent]: "Since Aiton's reference to Z. pumila excludes all the synonyms (and their type materials), Z. integrifolia may be treated as legitimate." Response. Though differently worded, this argument is a variant of Argument #3. Again, there is no requirement before 1958 that elements suitable for lectotypification be present. Had Linnaeus (1763) published Zamia pumila as he did but without inclusion of any references, the name would still be legitimate. The removal of his references by Linnaeus filius (1789) creates no reduced state of legitimacy. It is obvious that the failure of an author to designate a specimen that can serve as its type, or citation in his original materials of other publications in which such specimens may be referred, creates a significant uncertainty in assignment of the name to a definite taxon. The I.C.B.N. addresses this deficiency, by the process of neotypification (Art. 9.6). Where no specimen or suitable reference exists, the rules permit a specimen never seen by the author to be selected as a neotype. By this action a legitimate name that lacks clear meaning can be linked with a specimen and thereby be made precise. Phytologia (April 2009) 91(1) 101 Argument #6. "If the [lectotypic] element (Commelijn's t. 58 in this case) were to be included in any taxon published between 1762 [sic?] and 1980 (Z. integrifolia in this case), such an inclusion does not cause illegitimacy (Art. 52, Note 2). ...L. f. did not include this element for Z. integrifolia; therefore, the question of illegitimacy never arises." Response. The thrust of this argument is not entirely clear. The claim appears to be that by exclusion of the synonym the basis for the name was also excluded. This view was supported by reference to a rarely cited provision of the I.C.B.N., Art. 52.2, Note 2: "The inclusion, in a new taxon, of an element that was subsequently designated as the type of a name which, so typified, ought to have been adopted...does not in itself make the name of the new taxon illegitimate." The cited reference is irrelevant. A note as employed by the I.C.B.N. does not create a rule or restriction; it merely clarifies the meaning of the relevant Article. Plain reading of Note 2 creates no new content; it says merely that a special stated circumstance does not make the name illegitimate, though the implication is left that other circumstances may still do so. Argument #7. That the absence of known type material can be interpreted to mean there never was such material, in which event Zamia pumila would indeed be based on the cited references. "If there were evidence from the protologue of Z. pumila that there must have been original material, additional to that represented by the synonyms, then even if this material is no longer extant, I would agree that this situation would not meet the exclusionary requirements of Art. 52.2, and Z. integrifolia would be illegitimate. But...this has not been demonstrated." Response. This argument is the most interesting and potentially destabilizing of all offered. Whether or not Linnaeus had seen living or dried materials of the West Indian cycad is not known. He did not include the plant in his earlier (1737) treatment of plants he had studied at Hartecamp, Holland (in which his solitary cycad, later named Cycas circinalis, was placed between the palms Corypha and Phoenix). And following his death, no specimen was present in his herbarium (LINN). 102 Phytologia (April 2009) 91(1) Linnaeus, however, did not employ a single word from the phrase-names which he cited; his brief diagnosis was fully original. Nor was his epithet, pumila, of prior use. And none would claim that he saw nothing at Hartecamp other than those entities he knew well enough to describe at that time. Even his herbarium, between his death in 1778 and its arrival into the hands of Sir James Smith in 1784, suffered losses of many damaged sheets (Stafleu, 1971: 113). It thus cannot be ruled out that he may have been guided in whole or in part by materials no longer extant. Further, even if one were to assume the circumscription of Zamia pumila had been entirely fabricated, the I.C.B.N. does not provide for a distinction in treatment between such a baseless, illusory name and one whose type material had been lost. Nor does the I.C.B.N. require that evidence be provided that there had once been original material. Again, the logic is seductive that such a difference must call for different treatment. But in a real-world analysis it is impossible to document this distinction, and instability would be the only product of any effort to do so. Argument #8. That it is best to retain Zamia integrifolia because that name has been employed by some of the correspondents in the past. "Z. integrifolia was accepted in Flora of North America vol. 2 (1993: 348). ...[f the name is illegitimate, it needs to be conserved with a different type, for stability." Response. This proposal, aside from its implied lack of confidence by the correspondent, must be left to the judgment of other parties. CONCLUSION No arguments have been put forward in support of Zamia integrifolia that are firmly based on specific language of the I.C.B.N. None, it would appear, can stand in contravention to the clear language of Arts. 52.1 and 52.2, that an author's name is to be rejected if it was nomenclaturally superfluous when published, and that superfluity is caused by citation in synonymy of an earlier available name whose type Phytologia (April 2009) 91(1) 103 was not excluded by the author. Zamia integrifolia L. f. in Ait. must be interpreted under modern rules as a name that was illegitimate when published and is unavailable for use either in Florida or in the West Indies. But a cautionary note stands before unequivocal acceptance of Zamia floridana A. DC. as a replacement name for the Florida cycad. DeCandolle's name is preceded by a series of other binomials (Eckenwalder, 1980). Though none before Z. floridana is based on Florida materials, the taxon also occurs widely in the Bahamas and West Indies (Stevenson, 1987a). Should further investigation firmly assign one of these earlier names to Bahamian or West Indian materials of the Florida taxon, the Florida cycad may again require nomenclatural attention. ACKNOWLEDGMENTS This essay is a consequence of the outpouring of thoughts and judgments, both in support and in opposition, of James E. Eckenwalder, Kanchi Gandhi, Bruce F. Hansen, Walter S. Judd, John McNeill, Guy Nesom, Dan H. Nicolson, Bart M. Schulzman, Dennis W. Stevenson, Billie Turner, and Richard P. Wunderlin, to whom I am indebted. The e-mail file alone, transmitted through the courtesy of my colleague, W. S. Judd, though somewhat inflated by repetitions and occasional irrelevancies, has reached 18 pages of very small type. LITERATURE CITED Aiton, W. 1789. Hortus Kewensis 3: 478. London. Burch, D., D. B. Ward and D. W. Hall. 1988. Zamia, in: Checklist of the Woody Cultivated Plants of Florida. Institute of Food and Agricultural Sciences, Univ. of Florida, Gainesville. Publ. SP-33. 80 pp. Commelin, J. 1697. Horti Medici Amstelodamensis, pars prima. t. 58. Amsterdam. DeCandolle, A. 1868. Prodromus systematis naturalis regni vegetabilis 16(2): 544. Paris. 104 Phytologia (April 2009) 91(1) Eckenwalder, J. E. 1980. Taxonomy of the West Indian cycads. J. Arnold Arbor. 61: 701-722. McNeill, J., F. R. Barrie, H. M. Burdet, V. Demoulin, D. L. Hawksworth, K. Marhold, D. H. Nicolson, J. Prado, P. C. Silva, J. E. Skog, N. J. Turland and J. Wiersema, eds. 2006. The International Code of Botanical Nomenclature (Vienna Code), adopted by the 17th International Botanical Congress, Vienna, Austria, July 2005. Regnum Veg. 146: 1-568. Judd, W. S., C. S. Campbell, E. A. Kellogg and P. F. Stevens. 1999. Plant Systematics. Sinauer Assoc., Sunderland, Mass. 464 pp. Landry, G. P. 1993. Zamia, in: Flora N. Amer. 2: 347-349. Linnaeus, C. 1737. Hortus Cliffortianus, 482. Amsterdam. Linnaeus, C. 1763. Species Plantarum, ed. 2. 2: 1659. Stockholm. Stafleu, F. A. 1971. Linnaeus and the Linnaeans. Utrecht, Netherlands. Stevenson, D. W. 1987a. Again the West Indian zamias. Fairchild Trop. Gard. Bull. 42: 23-27. Stevenson, D. W. 1987b. Comments on character distribution, taxonomy, and nomenclature of the genus Zamia in the West Indies and Mexico. Encephalartos 9: 3-7. Stevenson, D. W. 1991. The Zamiaceae in the southeastern United States. J. Arnold Arbor. suppl. ser. 1: 367-384. Ward, D. B. 1968. Zamia, in: Checklist of the vascular flora of Florida, Part I. Fla. Agric. Exp. Sta. Bull. 726 (tech.). 72 pp. Ward, D. B., ed. 1979. Zamia, in: Rare and Endangered Biota of Florida, vol. 5: Plants. Univ. Presses of Florida, Gainesville. 175 Pp. Ward, D. B. 2001. New combinations in the Florida flora. Novon 11: 360-365. Wunderlin, R. P. 1998. Zamia, in: Guide to the vascular plants of Florida. Univ. Presses of Florida, Gainesville. 806 pp. Wunderlin, R. P. and B. F. Hansen. 2000. Zamia, in: Flora of Florida, vol. 1: Pteridophytes and Gymnosperms. Univ. Press of Florida, Gainesville. 365 pp. Phytologia (April 2009) 91(1) 105 MORUS MURRAYANA (MORACEAE): A NEW MULBERRY FROM EASTERN NORTH AMERICA Stephanie J. Galla, Brittney L. Viers, Paul E. Gradie, and Dayle E. Saar Department of Biological Sciences, Murray State University, Murray, KY 42071 USA dayle.saar@murraystate.edu ABSTRACT Mulberry trees (Morus: Moraceae) growing in relatively undisturbed, open woodland areas of western Kentucky exhibit exceptionally large leaves (blades often >15 cm long). Fruit size is also longer than reported for other species, and leaf vein patterns are unique. Field observations, combined with the use of herbarium specimens and molecular data warrant the establishment of a new species designation, Morus murrayana D.E. Saar and S.J. Galla (Murray State’s Mulberry). Phytologia 91(1):105-116 (April, 2009). KEY WORDS: Morus, Moraceae, mulberry, murrayana, Kentucky, internal transcribed spacer, ITS Mulberry trees (Morus L.: Moraceae) in western Kentucky and the surrounding states, identified as M. rubra L., were observed with exceptionally large leaves. In a search of plant keys (Jones, 2005; Mohlenbrock, 2002; Wunderlin, 1997; Swink & Wilhelm, 1994; Gleason & Cronquist, 1991; Radford et al., 1968; Steyermark, 1963; Britton & Brown, 1913) and detailed, authoritative books (Kurz, 2003; Dirr, 1998; Elias, 1987), only Wunderlin (1997) reported leaves of M. rubra over 15 cm in length. He listed the usual size as 7.5-18 cm but occasionally to 36 cm. Wunderlin assumed, based on his examination of herbarium specimens, that the large leaves were due to shade/sun forms or were perhaps associated with other growing conditions (pers. comm. to DES). It should be noted that leaf size is only mentioned in Wunderlin’s (1997) species description for M. rubra and he did not include this feature in the diagnostic characters given in the keys for 106 Phytologia (April 2009) 91(1) species identification. For the current study, the authors had the advantage of first-hand field observations of these trees growing in their natural habitat. Additional field data and DNA analysis demonstrate that this is a separate species from M. rubra and all others previously recognized. MORUS MURRAYANA D.E. Saar and S.J. Galla, sp. nov. Fig. 1. Arboles ad 20 m alto; folia alternatum, unifolius-quinquelobus, lamina ad 38 cm longus, serrulatus; fructus ad 4 cm longus, nigellus purpureus. TYPE: USA. KENTUCKY: Calloway Co. Frequent in open mesic woodlands dominated by Quercus spp. and Carya spp. along both sides of Watersport Rd. between gate to Racer Point and boat landing on Kentucky Lake, near Hancock Biological Station, Murray State University, ca. 25 km NW of Murray, KY (36° 43.87’ N; 088° 07.35’ W), 13 May 2006, Dayle E. Saar 3606 (Holotype: MUR; isotypes, BEREA, BRIT, EKU, F, MO, NCU, NY, TENN, US). Trees to 20 m tall with a single trunk, open crown vase-shaped to rounded. Sap milky. Bark on saplings smooth, medium brown with tan lenticels, becoming grayish-brown with very thin, long and narrow scaly plates. Winter buds with pseudoterminal present, dull brown, scales glabrous with minutely ciliated margins. Leaves alternate, simple; stipules light brown, membranous to 1.1 cm long and early deciduous; petioles 2.5-6 cm long; blades to 38 cm long, widest at or below the middle, unlobed to as many as five lobes, caudate at tip and oblique at base, serrate but occasionally double serrate, scabrous above and softly pubescent on veins and lamina below, pubescence on larger veins generally restricted to sides of veins, versus the dorsal peak (Fig. 1B); basal lateral veins larger than other laterals but smaller than the midvein, veins branching from two large lower laterals (tertiary) and other laterals from midvein (secondary) curve towards the tip as they approach the blade margin without entering a tooth and only the finest veins end in a tooth (Fig. 2C). Inflorescences of unisexual flowers axillary on short peduncles; individuals varying from monoecious to polygamodioecious to dioecious. Fruit multiple, blackish purple with dark reddish-purple juice at maturity, to 4 cm long and 1.5 cm wide but often thinner, with much variation in size on a single individual. The Phytologia (April 2009) 91(1) 107 Figure 1. Morus murrayana with mature fruit showing variation in leaf morphology (leaves not at maximum size when fruit begins to mature); A. abaxial surface; B. adaxial surface. Scale bar = 5.0 cm. 108 Phytologia (April 2009) 91(1) specific epithet, murrayana, was chosen to honor Murray State University, Murray, KY (Murray State’s Mulberry). Figure 2. Leaves of A. Morus rubra; B. M. alba; and C. M. murrayana showing differences in veining patterns. Leaves not drawn to scale with each other. See text for further descriptions. GENERAL MORPHOLOGY AND NATURAL HISTORY Many individuals of Morus murrayana were observed growing in wet-mesic to dry-mesic open woodlands, in partly shaded to sunny locations; the few individuals located in heavy shade were growing poorly. They are virtually absent where Red Maple (Acer rubrum L.) has come in under overstories dominated by oaks (Quercus L. spp.) and hickories (Carya Nutt. spp.) and completely filled-in the canopy. MM. murrayana occurs in natural as well as mildly disturbed localities, but generally is not in high disturbance places such as fence rows. Individuals observed for this study ranged in age from saplings to large trees; none were stump-sprouts. In other words, no correlation Phytologia (April 2009) 91(1) 109 has been detected between the large leaves and the trees’ age, condition, and/or habitat including soil type. The morphological extremes in blade shape can be observed on the same branch next to each other. However, almost all trees produce either >80% lobed leaves or are >80% unlobed; few individuals display anything that approaches equal proportions of lobed and unlobed leaves. In fall, the leaves turn medium yellow, the smaller internal leaves falling first. Trees produce either predominately staminate or carpellate inflorescences, but the presence of some staminate inflorescences on carpellate trees and vise-versa is common, especially on large saplings and older individuals. Both staminate and carpellate inflorescences may occur on the same large branch, usually separated on different twigs. This is in contrast with M. rubra, which is monoecious (Elias, 1987; Jones, 2005). Fruit ripens in western Kentucky during June and early July. Morus murrayana has been sighted by one or more of the authors in Kentucky, Tennessee, Missouri, and Illinois. Herbarium specimens were also used to locate additional occurrences. Specimens of M. murrayana were readily recognizable among the accessions based on overall leaf size, caudate blade tips, and the venation pattern. Specimens confirm that this species has existed or currently exists in the four states personally documented by the authors, plus an additional six states that include Indiana, Mississippi, Louisiana, Virginia, North Carolina, and Alabama. The oldest specimen of M. murrayana (identified as M. rubra) was collected in the southern Illinois city of Carbondale in 1870. Other historical specimens date back to 1889, 1919, and 1937, in addition to those collected more recently. None of the label descriptions indicate a disturbed habitat, although habitat details were omitted on many labels. Most large-leafed specimens were identified as M. rubra, presumably due to the similar pattern of leaf pubescence. However, these trees were clearly problematic to taxonomists. Some specimens were identified as M. alba L., a few were listed as M. rubra x M. alba, and some had notations of “Morus sp.” or “could not be determined.” Many had annotations different than the original label identification (MZ. alba to M. rubra and vise-versa). 110 Phytologia (April 2009) 91(1) OTHER SPECIES OF MORUS IN NORTH AMERICA Morus consists of about 12 species (Mabberley, 1997), only two of which (M. rubra and M. microphylla Buckley) are native to the United States, Canada, and Mexico; other species are native to Asia with some ranges extending west into Europe. M. microphylla is a small shrub or small tree with leaves to 5 cm in length. M. murrayana is most often confused with M. rubra. It can be distinguished from M. rubra based on leaf vein pattern (Fig. 2), leaves longer than 15 cm with caudate tips (vs. leaves <15 cm with cuspidate to broadly acute or acuminate tips), and fruits longer than 3 cm (vs. <3 cm). Another species, Morus alba, is naturalized from Asia and is widespread in many areas of North America, as a result of escapes from cultivation as a street planting and from an unsuccessful attempt in the 1830s to establish a silk industry in the United States (Federico, 1997), as the leaves are used to feed larvae of the silkworm, Bombyx mori L. M. alba has leaves that are shiny above and pubescence below is either absent or scattered in vein axils or sparse along the larger veins. Morus nigra L. and M. alba var. multicaulis (Perr.) Loudon (syn. M. multicaulis Perr.) also have been documented in the US (Wunderlin, 1997; Jones, 2005). Unlike M. murrayana, M. nigra has dark brown bark and elliptic fruit to 2.5 cm long and wide (H. Sang, 2003). Further, Wunderlin (1997) reports that M. nigra, occasionally cultivated in North America, is not known to naturalize. M. alba var. multicaulis has leaves to 30 cm (L. Sang, 2003). However, the blades are thick and wrinkled, which does not describe MZ. murrayana, and the pattern of restricted pubescence below and glabrous above is consistent with M. alba (not M. murrayana). Also, live specimens of M. murrayana show no tendency for multiple trunks. Plants of Morus rubra and M. alba are known to hybridize and produce intermediate pubescence patterns on leaves, but they did not produce large leaves and fruit (Burgess et al., 2005). MM. murryana was not observed with M. rubra, but in the few instances where it occurred in the vicinity of M. alba, intermediates were not present. Phytologia (April 2009) 91(1) 111 MOLECULAR ANALYSIS AND COMPARISONS Materials and Methods: The entire herbarium collection of Morus at the Missouri Botanical Garden (MO) was inspected for similar leaf and fruit characteristics by DES. Sequences for the internal transcribed spacer region (ITS) of nuclear ribosomal DNA (nrDNA) are available in GenBank (www.ncbi.nlm.nih.gov) for the species of Morus native to the Eastern Hemisphere. For this study, DNA was extracted from two individuals of M. rubra and three of M. murrayana using Quagen DNeasy kits. Amplification was carried out following the protocol detailed in Saar et al. (2003). DNA was sequenced in the DNA Core Facility at Northern Illinois University, DeKalb, Illinois, on a Beckman-Coulter capillary sequencer. All sequences of M. rubra and M. murrayana were aligned with Clustal X software (Thompson et al., 2003). There were differences between the sequences of M. rubra and M. murrayana, but there was no intraspecific variation. Therefore, only one sequence of M. murrayana was necessary for comparison to existing GenBank accessions using a BLAST search (Altschul et al., 1990). The closest matches from GenBank, together with the new sequences of M. rubra and M. murrayana, were also aligned in Clustal X. Results: There were no herbarium specimens at MO with similar morphological characteristics from any continent, except those identified as Morus rubra. The BLAST search indicates the closest species to M. murrayana are M. macroura Miq., M. lhou Koidz., M. bombycis Koidz., M. cathayan Hemsl., M. atropurpurea Roxb., M. alba, M. nigra, M. australis Poir., and M_ multicaulis. Nucleotide polymorphisms are summarized in Fig. 3. DISCUSSION From the list of species whose sequences most closely match M. murrayana (Fig. 3), Morus atropurpurea, M. bombycis, M. lhou, Phytologia (April 2009) 91(1) 112 ‘soloods 104}0 9UO puk DU_ADAINUU SI 9Seq oNbiuN 4 ‘ouDADLINU PF SI dSeq onbIUN , :Sd}0N “$79 OSE Je SUIBOQ BUDS S97 ‘Zp OSRq Je SUISOq Z SLI ‘17 98eq Je suIdoq DUDS Gg'¢ ‘EG OSeq Je SUISOQ | SI] ‘WUoOWUSI]e sy) UT “sUOISsoDNR YURGUOH WOY sidyjO ‘Apnys sty} Woy vunddsiniu Jy pure vAqn4 PW JO sodUsnbog “YURGUdH Jo YoIkOS [SVT B UO poseq ‘punADLUNUL SNAOP OJ SIQUINbIS yS9SO]I WoO soseq pousIye 179 Wo swsiydiowAjod opyoojonu o[8uls jo Arewmwng *¢ sandy : HK KKK KKK KKK KERR KKK KKH KKK HK > S8ION 7--990028323395-333.1093533905063 ee0656359---65 (euexkerrnu STSSO9rd OLL--999999--OWOOLO-W9090 (eerndindo7z3e GPUISPEAY DLL-99-9099--OWO9LO-wo09 (ezqnt 9TSSo9rd DLL9DN999999--DWODLO-wOd9 (STTNeOTIT AW €007TPONY DLL-9999999--DWODL9-wo09 (eznorOeW O007PONW DLL-0809999--DVWODLO-W999 (ez6Tu ZOOZPONW ODLL-90999999--OWOOLO-W909 (eqTe 8661 PONY DLL-9999099--DWOOLO-W909 (NOYT 666 T PONY ODLL-0999099--dvwo0ee-VO09 (sTTezqsne POOCPONY DLL-9999999--OWOOLO-W999 (euehey eo TOOZ PONY OLL-9999999--DWOOLO-W999 (sTOAquoq 9007PONY GLODEESTETBITOEPCICESGO LOG HEC IOG8LOSHECBLIO9 1 (setoeds) ‘on yuequey! TOOOO66LPEG6LLZTOSZE6EZTTIES88888LLLLLLLLOSEEZO 99999SSSSSDPPPPPEETZZZI10000000000000000000 -(*up Sutpesez) zequmy eseg Phytologia (April 2009) 91(1) 113 and M. multicaulis are synonyms or varieties of M. alba (Ghafoor, 1985; Shu, 2003; Index Kewensis). All species from this list can be separated from M. murrayana based on morphology. M. macroura has yellowish-white fruit when mature, 6-12 cm long (Ghafoor, 1985; N. Sang, 2003). (M. murrayana fruits are blackish-purple and < 4.0 cm.) The Trade Winds Fruit Company (www.tradewindsfruit.com) reports that M. macroura is only hardy to about 18-25°F (varies by individual). Western Kentucky and southern Illinois are in USDA zone 6 (average low of -10-0°F). M. cathayana has leaves 8-20 cm long, but they are thick and papery and winter buds are white pubescent (S. Sang, 2003), again, unlike M. murrayana. Morphologic distinctions from M. alba and M. nigra have been discussed previously. The fruits of M. australis are <2.5 cm long and plants are shrubs or small trees (Shu, 2003; J. Sang, 2003) with twisted branches (Dirr, 1998). In addition to M. alba, ten other species from Asia are described in Flora of China (Shu, 2003). All species and varieties have leaves < 15 cm except for M. nigra with 6-12(-20) and M. cathayana with 8-20 cm leaves (both discussed previously). Four species are included in the Flora of Pakistan. One of these species, M. serrata Roxb. (syn. M. alba var. serrata (Roxb.) Bureau), which is confined to the Indo-Pakistan subcontinent, has leaves 5-15 cm long. However, this species has longer stipules (1.5-2.5 cm long) and smaller fruits (0.8-2.5 cm long) (Ghafoor, 1985) than does M. murrayana. The DNA sequence data separate M. murrayana from all others in the GenBank database. Sequences of M. rubra and the Asian species are similar, whereas M. murrayana has a notable 13-base insertion plus five 1-2 base indels and 16 single base substitutions (Fig. 3). It is clearly the most distinctive sequence of the entire group. Although it is somewhat unusual to describe a new, wide- ranging, tree-sized species from the US, it is not without precedent. Due in part to the fact that there are so few species of Morus found in North America, very few characters are required to separate them. For example, the most common sympatric species ranges include M. rubra and M. alba, which can be separated based on leaf pubescence, thereby eliminating the need to elaborate with additional descriptions. With so few diagnostic characters utilized, M. murrayana falls within the 114 Phytologia (April 2009) 91(1) parameters of virtually all plant keys for this genus in North America and falls under M. rubra. Thus, skepticism may be minimal because it is seemingly a good “fit” with the key. We have not observed M. rubra growing with M. murrayana, which could have facilitated direct comparisons. Further, with only two native species, Morus is not a particularly attractive candidate for taxonomic study in North America. The reputation of our native M. rubra may suffer due to a close resemblance with its weedy, non-native congener, M. alba. Further studies are underway to learn more about the natural history of this species, and to produce a more detailed and extensive distribution map. Key to Native and Introduced Species in North America: 1. Leaves 2-5 cm in length, strongly bicolored (dull dark green above, pale green below); shrubs or small straggly trees to 7 m; trees of the American SW and N Mexico. . . . M. microphylla 1. Leaves 3.8-14 cm long or longer, not strongly bicolored; trees. . . 2 2. Leaves glabrous above and often lustrous, glabrous below or pubescence restricted to scattered hairs in vein axils or scattered along larger veins; mature fruit white through pink to'blackish purple. 5) swcy,t. eee. 1 SAM oro M. alba 2. Leaves scabrous on upper surface and soft pubescent below. . 3 3. Leaves with cordate bases; fruits elliptic to long ovoid, < 2.5 cm long, maturing from red to black; landscape plant... . . M. nigra 3. Leaf bases variously oblique to slightly cordate; fruits cylindrical and >2.5 cm long; mature fruits blackish purple... . . 4 4. Leaves to 15 cm long but often <10 cm, acute to acuminate at tip, lateral veins (secondary) above lowest lateral fairly straight and ending in a tooth; mature fruit to 3 cm long... . . M. rubra 4. Leaves to 38 cm long, outer three leaves on branchlets almost always > 15 cm, caudate, lateral veins curve before reaching margins, only tiniest veins end in a tooth; mature fruit to 4 cm long and 1.5 cm wide but often thinner, with much size variation on a single individual. . M. murrayana Phytologia (April 2009) 91(1) 115 ACKNOWLEDGEMENTS The authors thank Andrew M. Saar for the two technical drawing figures and helpful observations on leaf venation patterns, the Kentucky Academy of Science for funds to SJG and DES, and the faculty Committee on Institutional Studies and Research at Murray State University for funds provided to DES. W. Scott Grayburn sequenced the DNA at NIU. Michael O. Saar assisted DES with the inspection of specimens at MO. Two reviewers provided very helpful comments on an earlier version of the manuscript. In addition to the Murray State Univ. Herbarium (MUR), we are grateful for specimens borrowed from the following herbaria: Missouri State Univ. (SMS), Northern Kentucky Univ. (KNK), Southeast Missouri State Univ. (SEMO), Southern Illinois Univ. (SIU), and the Univ. of Missouri, Columbia (UMO). LITERATURE CITED Altschul S.F., W. Gish, and W. Miller. 1990. Basic local alignment tool. J. Mol. Biol. 215: 403-410. Britton, N.L. and A. Brown. 1913. An illustrated flora of the northern United States and Canada, ed. 2. Dover reprint (1970), Dover Publications, Inc., New York, NY. Burgess, K.S., M. Morgan, L. Deverno, and C. Husband. 2005. Asymmetrical introgression between two Morus species (M. alba, M. rubra) that differ in abundance. Mol. Ecol. 14: 3471-3483. Dirr, M.A. 1998. Manual of woody landscape plants, ed. 5. Stipes Publishing L.L.C., Champaign, IL. Elias, T.S. 1987. The complete trees of North America field guide and natural history, Gramercy Publishing Co., New York, NY. Federico, G. 1997. An Economic History of the Silk Industry, 1830- 1930, (Cambridge Studies in Modern Economic History) Cambridge University Press, New York, NY. Ghfoor, A. 1985. Moraceae. Flora of Pakistan, vol. 171, Missouri Botanical Garden Press, St. Louis, MO. Gleason, H.A. and A. Cronquist. 1991. Manual of vascular plants of northeastern United States and adjacent Canada, ed. 2. The New York Botanical Garden, Bronx, NY. 116 Phytologia (April 2009) 91(1) Jones, R.L. 2005. Plant Life of Kentucky. University Press of Kentucky, Lexington, KY. Kurz, D. 2003. Trees of Missouri, C. Overby, ed. Missouri Department of Conservation, Jefferson City, MO. Mabberley, D.J. 1997. The plant-book, ed. 2. Cambridge University Press, New York, NY. Mohlenbrock, R.H. 2002. Vascular flora of Illinois. Southern Illinois University Press, Carbondale, IL. Radford, A.E., H.E. Ahles, and C.R. Bell. 1968. Manual of the Vascular Flora of the Carolinas. University of North Carolina Press, Chapel Hill, NC. Saar, D.E., N.O. Polans, and P.D. Sorensen. 2003. A phylogenetic analysis of the genus Dahlia (Asteraceae) based on internal and external transcribed spacer regions of nuclear ribosomal DNA. Syst. Bot. 28: 627-639. Sang, H. 2003. Morus nigra. Flora of China, vol. 5, Missouri Botanical Garden Press, St. Louis, MO. Sang, J. 2003. Morus australis. Flora of China, vol. 5, Missouri Botanical Garden Press, St. Louis, MO. Sang, L. 2003. Morus alba. Flora of China, vol. 5, Missouri Botanical Garden Press, St. Louis, MO. Sang, N. 2003. Morus macroura. Flora of China, vol. 5, Missouri Botanical Garden Press, St. Louis, MO. Sang, S. 2003. Morus cathayana. Flora of China, vol. 5, Missouri Botanical Garden Press, St. Louis, MO. Shu, S. 2003. Morus. Flora of China, vol. 5, Missouri Botanical Garden Press, St. Louis, MO. Steyermark, J.A. 1963. Flora of Missouri. Iowa State University Press, Ames, IA. Swink, F. and G. Wilhelm. 1994. Plants of the Chicago region, ed. 4. Indiana Academy of Science, Indianapolis, IN. Thompson, J.D., T.J. Gibson, F. Plewniak, F. Jeanmougin, and D.G. Higgins. 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25: 4876-4882. Wunderlin, R.P. 1997. Morus [Moraceae] in Flora of North America north of Mexico, vol. 3, pp. 390-392, Flora of North America Editorial Committee, eds. Oxford University Press, New York, NY. Phytologia (April 2009) 91(1) 117 ANALYSES AND TAXONOMIC UTILITY OF THE CEDARWOOD OILS OF THE SERRATE LEAF JUNIPERS OF THE WESTERN HEMISPHERE Robert P. Adams Biology Department, Baylor University, Box 727, Gruver, TX, 79040 Robert_ Adams@baylor.edu ABSTRACT Analyses of the wood oils of the serrate leaf margined Juniperus of the western hemisphere (21 species, 3 varieties and 1 form) are presented. All taxa have considerable amounts of cedrol, widdrol, cis-thujopsene, a-cedrene and f-cedrene. In general, there was little correlation between cedarwood oil compositions and phylogeny in this section of Juniperus. Phytologia 91(1):117-139 (April, 2009). KEY WORDS: Juniperus, Cupressaceae, wood oils, taxonomy, serrate leaf, cedrol, widdrol, cis-thujopsene, a-cedrene and B-cedrene. The serrate leaf margined junipers of the western hemisphere appear to represent a natural sub-group of Juniperus (Adams, 2008). A phylogenetic tree (Fig. 1) shows the relationships among these junipers based on nrDNA and trnC-trnD sequences. These junipers evolved at the margins of deserts in the southwestern US and Mexico. The southwestern US - Mexico, the northern-Mediterranean, and central- Asia - western China are the three centers of biodiversity of Juniperus. The serrate leaf junipers appear to be the most recent species of the genus (Adams, 2008). The group is composed of: J. angosturana R. P. Adams, J. arizonica (R. P. Adams) R. P. Adams, J. ashei Buchholz, J. a. var. ovata R. P. Adams, J. californica Carriere, J. coahuilensis (Martinez) Gaussen ex R. P. Adams, J. comitana Martinez, J. compacta (Mart.) R. P. Adams, J. deppeana Steudel var. deppeana, J. d. forma elongata R. P. Adams, J. d. forma sperryi (Correll) R. P. Adams, J. d. forma zacatacensis (Mart.) R. P. Adams, J. d. var. gamboana (Mart.) R. P. Adams, J. d. var. patoniana (Martinez) Zanoni, J. d. var. robusta Martinez, J. durangensis Martinez, J. flaccida Schlecht., J. grandis R. 118 Phytologia (April 2009) 91(1) P. Adams, J. jaliscana Martinez, J. martinezii Perez de la Rosa, J. monosperma (Engelm.) Sarg., J. monticola Martinez forma monticola, J. m. forma orizabensis Martinez, J. occidentalis Hook., J. osteosperma (Torr.) Little, J. pinchotii Sudworth, J. poblana (Martinez) R. P. Adams, J. saltillensis M. T. Hall, and J. standleyi Steyermark. serrate Juniperus w. hemisphere californica Baysian tree nrDNA + tmC-tmnD monosperma angosturana coahuilensis pinchoti arizonica occidentalis grandis osteosperma saltillensis compacta deppeana d. var. patoniana d. f. zacatacensis d. var. robusta d. vat. gamboana ashei comitana poblana Standley jaliscana monticola flaccida durangensis martinezii Figure 1. Phylogenetic tree of the serrate junipers (from Adams, 2008). Although the leaf essential oils of Juniperus have been extensively utilized for taxonomic purposes (Adams, 1991a; Adams, 2008), the wood oils have not received much taxonomic attention. Adams (1987, Adams, 1991b) examined the wood oils of junipers from the United States as potential sources of cedarwood oil and reviewed the literature on early analyses of Juniperus wood oils. Commercial cedarwood oils have been obtained from 3 genera of Cupressaceae: Juniperus (Texas, Virginia and African oils); Cupressus (China) and Cedrus (Morocco, India) according to Bauer and Garbe (1985). However, Texas (Juniperus ashei Buch.), Virginia (J. virginiana L.) and Chinese (putatively, Cupressus funebris Endl.) cedarwood oils account for almost all the cedarwood oil commercially produced today (Lawrence, 2003). The heartwood oils of the Cupressaceae are well known for having the same components across the family (i.e., evolutionally conserved), so the occurrence of similar oils in different genera is not surprising. It is probably due to the conservation of the principal commercially important components (cedrol, widdrol, cis-thujopsene, a-cedrene and Phytologia (April 2009) 91(1) 119 B-cedrene) that the wood oil compositions have not been utilized for taxonomic purposes. A second reason that wood oils have not been widely utilized is the difficultly in taking samples. Leaf sampling does not harm a tree. But to obtain a wood sample requires cutting down the tree, cutting off a limb, or taking a coring sample. Taking tree cores is the least destructive, but presents a problem if steam distillation is utilized to obtain the wood oil, as a core sample consists of only a few grams of wood and the oil can easily be lost on the walls of the steam distillation apparatus. Comparison of steam distillation versus solvent extraction using wood from the same tree (J. ashei) is shown in Table 1. Notice that the highest yield was obtained from steam distilled wood shavings and that 24 h of pentane extraction of wood chips removed only about one- half of the oil obtained by steam distillation of wood shavings. Using finely ground wood, resulted in about the same yield as using wood chips. Table 1. Comparison using J. ashei_wood for yields (oven dry wt basis) and the concentrations of key components of oils obtained by steam distillation (24 h), vs. various wood chip sizes and extractions with pentane . ee a ae steam Pentane extractions distilled | woodchips wood from’ ground wood variable shavings’ ext'd24h’ _re-ext'd +72h’ ext'd 24h* per cent yield 3.7% 1.6% 1.6% 1.6% a-cedrene 7.0 2.4 4.2 1.8 B-cedrene iy Int 2.0 1.0 cis-thujopsene 14.6 5.6 10.8 Did cedrol 48.1 64.3 66.1 47.2 cis-thujopsenic acid 0.6 10.4 4.8 LI ‘shavings obtained from original wood, then steam distilled 24 h. *wood cut into 25 mm x 3 mm x 3mm wood chips, pentane extracted by shaking, 24 h. (# 9696) *wood from Ist 24 h extraction (*), then the extracted wood chips were ground in a coffee mill and a second, 72h, pentane (# 9697) performed. “original sample wood ground in coffee mill, then pentane extracted pentane for 24 h (#9700). 120 Phytologia (April 2009) 91(1) None of the pentane extractions gave exactly the same results as the steam distillation (Table 1). However, steam distillation can result in decomposition (Adams, 1991b), whereas solvent extraction is a very gentle method. This is shown in the marked increase of cis-thujopsenic acid in the solvent extracts (Table 1). Free acids may be dissolved in the steam condensate and return to the boiling chamber or they may decompose during distillation (Adams, 1991b). If all the wood samples are extracted in the same manner (ex. 24 h, shaking in pentane, cut to uniform sizes), solvent extraction should produce a reasonable snapshot of the profile of the wood oils. The purpose of this paper was to present analyses of the wood oils of all the serrate junipers of the western hemisphere and evaluate these data for use as taxonomic characters. MATERIAL AND METHODS Samples used in the study: J. angosturana, 10.5 km e of Villa Juarez (road from Cerritos to Rio Verde), thence s 1.3 km, San Luis Potosi, Mexico, Lab # 9743 Adams 8714, J. arizonica, Rock Hound St. Park, Luna Co., NM, Lab # 9725, Adams 7637, J. ashei, 1.6 km e of Llano R., on I10, east of Junction, Kimble Co., TX, Lab #9721., Adams 5010, J. californica, 13 km n of 140 on road to Kelso, San Bernardino Co., CA, Lab # 9750, Adams 5071, J. coahuilensis, 32 km n of Alpine, TX, Jeff Davis Co., Lab # 9723, Adams 4994, J. comitana, 14 km s of Comitan and thence 14 km e on rd to Montebello, Chiapas, MX, Lab # 9737, Adams 6862, J. compacta, near the summit of Cerro Potosi, Nuevo Leon, MX, Lab # 9742, Adams 6898, J. deppeana var. deppeana, 32 km nw of Ft. Davis on Tex 118, Jeff Davis Co., Lab # 9744, Adams 4983, J. d. var. gamboana, 17 km n of Comitan on Mex. 190, Chiapas, MX, Lab # 9735, Adams 6864, J. d. var. patoniana, km 152 on Mex. 40, 52 km w of El Salto, Durango, MX, Lab # 9738, Adams 6838, J. d. var. robusta, west of Creel, Chihuahua, MX, Lab # 9728, Adams 6826, J. d. forma zacatacensis, 18 km w of Sombrerete, between km 178 & 179 on Mex. 45, Zacatecas, MX, Lab # 9740, Adams 6840, J. durangensis, nw side of Mex. 40, km 152, 52 km w of El Salto, Durango, MX, Lab # 9749, Adams 6832, J. flaccida, 20-25 km e of San Roberto Jct., on Mex. 60, Nuevo Leon, MX, Lab # 9745 Phytologia (April 2009) 91 (1) 121 Adams 6892, J. grandis, at Sonora Bridge Campground Rd., 2 km w of Jct of CA 108 and US 285 on CA 108, Mono Co., CA, Lab # 9734, Adams 5061, J. jaliscana, 19 km e of Mex. 200, on road to Cuale, Jalisco, MX, Lab # 9739, Adams 6846, J. martinezii, on La Quebrada Ranch, 40 km n of Lago de Moreno off Mex. 85 to Amarillo, thence 10 km e on dirt rd to La Quebrada Ranch, Jalisco, MX, Lab # 9727, Adams 8709, J. monosperma, 1.6 km w of Santa Rosa, on 140, Guadalupe Co., NM, Lab # 9748, Adams 5028, J. monticola forma monticola, 1 km n of jct of Mex. 105 and El Chico Natl. Park, on road to El Chico Natl. Park (8 km ne of Pachuca), Hidalgo, MX, Lab # 9747, Adams 6874, J. occidentalis, 58 km w of Juntura on US 20, Malheur Co., OR, Lab # 9724, Adams 5085, J. osteosperma, 25 km e of Monticello, on US 666, San Juan Co., UT, Lab # 9741, Adams 5053, J. pinchotii Sudw., 10 km w of Sheffield, on 110, Pecos Co., TX, Lab # 9722, Adams 5004, J. poblana, at KM 62 on Mex. 190, 62 km s of Oaxaca, Oaxaca, MX, Lab # 9729 Adams 6871, J. saltillensis, 14 kme of San Roberto Jct., on Mex. 60, Nuevo Leon, MX, Lab # 9726, Adams 6887 and J. standleyi, 24 km nw of Huehuetango on road to San Juan Ixcoy (s of El Oro), Guatemala, Lab # 9746, Adams 6852. Vouchers are in the herbarium, Baylor University (BAYLU). Wood samples were radially cut in 1 cm segments using a band saw. The radial sections were then cut linearly into 2 x 5 mm (x 1 cm) pieces. The wood (25 g) was placed in a 125 mL screw cap bottle to which 50 mL of pentane was added. The bottles were shaken for 24 h on a rotary shaker. The pentane extract was filtered and the pentane evaporated by use of nitrogen. The extracted wood was oven dried 48 h, 100 °C for use in the oven dry weight calculations. Percent yields were determined on an oven dry weight basis as: 100 x oil wt./(oil wt. + oven dry wood wt.). All oil samples (including commercial cedarwood oils) were dissolved in diethyl ether (10% oil solution) and stored at - 20°C until analyzed. The extracts were analyzed on a HP5971 MSD mass spectrometer operated in the EI mode, scan time Isec., acquisition mass range: 41-500, directly coupled to a HP 5890 gas chromatograph, using a J & W DB-5, 0.26 mm x 30 m, 0.25 micron coating thickness, fused silica capillary column, 0.2 wL injected of a 10 % solution in 122 Phytologia (April 2009) 91(1) diethyl ether, and split 1/10, injector: 220 °C, transfer and MSD: 240 °C, column temperature linearly programmed: 60° - 246 °C/ 3 °C min. Identifications were made by library searches of our volatile oil library (10), using the HP Chemstation library search routines, coupled with retention time data of authentic reference compounds. Quantitation was by use of the HP Chemstation software. Normally one would report the data as FID values, but considering the difficultly of the peaks overlapping such that quantitation involved using single ion chromatograms to estimated the concentrations (eg. cedrol/ widdrol, etc.), it was not practical to quantitate the components by GC-FID. RESULTS AND DISCUSSION Tables 2 and 3 show the complete analyses of the cedarwood oils of all 25 taxa of serrate leaf margined junipers of the western hemisphere. Due to the use of a liquid extraction, considerable amounts of cis-thujopsenic acid (and other acids) were present in the extracts. It appears much of the cis-thujopsenic acid is degraded or left in the water condensate during steam distillation (Table 1). Although most taxa have considerable amounts of cedrol, widdrol, cis- thujopsene, a-cedrene and f-cedrene, there are many un-identified sesquiterpenoids. However, it is surprising to find such a large number of un-identified compounds. Often, an un-identified compound was present in only one or twice taxa. It appears that there may be considerable synthesis of non-specific products in the wood. Keeling and Bohlmann (2006) discuss the defense nature of terpenoids and note that maintaining a diverse array of chemicals may be effective as a plant defense. The major components useful in commercial cedarwood oils are cedrol, widdrol, cis-thujopsene, a-cedrene and f-cedrene. Table 4 shows the compositions of these five constituents in the 25 taxa of this study. Notice that even in phylogenetically similar taxa (Figure 1, arizonica, occidentalis, grandis and osteosperma), there is considerable variation in the amounts of these components. In fact, it seems that there is as much variation among these presumably closely related species as among other more distantly related species (Table 4). Phytologia (April 2009) 91(1) 123 The sum of cedrol, widdrol, cis-thujopsene, a-cedrene and f- cedrene gives one some indication of the oils’ utility as cedarwood oil. The sums range from 22% of the total oil (/. jaliscana, Table 4) to 70.4% (J. angosturana, Table 4). The species with low sums generally have considerable amounts of unknown compounds. The percent yields ranged from 0.04% (J. deppeana var. patoniana) to 3.4% (J. standleyi). As a reference, the source of Texas cedarwood oil, J. ashei, had a 1.7% oil yield. The product of the sum of the key components x % yield (S x %, Table 4) is an index to the relative commercial potential of a species. This index varied from 2.0 (J. deppeana var. patoniana) up to 169.4 for J. angosturana with J. ashei having an index value of 104.9. Although J. angosturana (169.4), J. d. var. gamboana (128.6) and J. standleyi (143.1), all from Mexico, scored higher than J. ashei from the United States (mostly Texas), they are generally not found in large enough populations to sustain continued harvest of trees for cedarwood oil. Juniperus arizonica, J. californica, J. grandis, J. monosperma, J. occidentalis and J. pinchotii are weedy, widespread junipers of the western US but these are either lacking a high concentration of the key compounds or their percent yields are low and do not appear suitable for commercial cedarwood oil production. In conclusion, the cedarwood oils in this section of Juniperus do not seem to be useful to taxonomy at the specific level. It would be interesting to examine geographic variation within a species to determine if the wood oils might be useful for populational studies. ACKNOWLEDGEMENTS Thanks to Tom Zanoni and Glenn Harris for assistance on the field trips in Mexico and Guatemala. This research supported in part by funds from Baylor University. 124 Phytologia (April 2009) 91(1) LITERATURE CITED Adams, R. P. 1987. Investigation of Juniperus species of the United States for new sources of cedar wood oil. Econ. Bot. 41: 48-54. Adams, R. P. 1991a. Analysis of Juniper and other forest tree oils. pp. 131-157. In: Modern Methods of Plant Analysis, New Series: Oil and Waxes. H.-F. Linskens and J. F. Jackson, eds. Springler- Verlag, Berlin. Adams, R. P. 1991b. Cedarwood oil - Analysis and properties. pp. 159- 173. In: Modern Methods of Plant Analysis, New Series: Oil and Waxes. H.-F. Linskens and J. F. Jackson, eds. Springler- Verlag, Berlin. Bauer, K. and D. Garbe. 1985. Common fragrance and flavor materials. CVH Verlagsgesellschaft, Weinheim, Germany. Keeling, C. and J. Bohlmann. 2006. Genes, enzymes and chemicals of terpenoid diversity in the constitutive and induced defence of conifers against insects and pathogens. New Phytologist 170: 657-675. Lawrence, B. M. 2003. Essential Oils 1995-2000. 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FOO, 0 c0 ues } ide yuIOD oYse Wiesp golp JOUISNLIOJ-sueI} ]01e}0}-SsuRy jouuewuidoseiepues ploe s1louvsape}90 ploe 19/0 PET EOT/TOT “Ep Suadsoynbsas PET SOL 6P1 ‘6S [ediayinbsas OST ISI EPSET plow” ynbsoas Phytologia (April 2009) 91(1) 139 Table 4. Comparison of the major oil components among taxa. ACDR = a-cedrene, BCDR = -cedrene, THJP = cis-thujopsene, WDRL = widdrol, CDRL = cedrol, CTJA = cis-thujopsenic acid. %yld = % yield on oven dry wt basis (oil/(extracted wood, dried 48h, 100°C). ACDR_ BCDR_THJP WDRLCDRL Sum %yld Sx % J. californica 1.2 DG 22s py AMD, MAT GSA: O.2 5.0 J. monosperma 0.7 0.5 SO ATA GTA eS less 'G:3 9.4 J. angosturana 1.4 O7 17S t 510° 70.6 24 169.4 J. coahuilensis tt 0.5 18.4 - 7.4 | Se. Bib 59.8 J. pinchotii 3.6 16 120. 42. 386 600 0.2 12.0 J. arizonica 1.9 0.6 . (145 o 76 B.L a277. 0.6 16.6 J. occidentalis 0.9 0.8 6.4 43.3 514 0.5 25.7 t J. grandis 1.4 0.9" 154 - ALS Spo, 0.2 11.9 J. osteosperma 1.3 O97. 13.7 t 36.3 ° 520 O35 15.6 J. saltillensis {2 0:7 4 4)°930 t 46.9 57.8 0.1 5.8 J. compacta 0.4 0.4 25 - co: Fo alae pes lam 513 J. deppeana 7.6 2.0 9.1 12.0. 25.49" (564° “OS 16.9 patoniana 5 0.8 7.22 100 30.4 49.9 0.04 2.0 zacatacensis 4.1 14 WalOsr, AGIOS HES es3 ce NOs 16.6 robusta 3.9 0.9 G3 15-6151 200287) 7 20.1 gamboana 2.5 VO. 8 ON 55.2 ~C43 2:0, 128.6 J. ashei 0.6 04) 1200 120) “36:7 S67. 1.7 1048 J. comitana 0.5 0.2 2.9 - 43.2 468 1.0 46.8 J. poblana a 0.9 G6.~ 206) 21-0 9352.4 1-8 94.3 J. standleyi 2.6 2 “IOS. 252° 2:8 "a2. ssa Fast J. jaliscana 2.6 Ot 1G 160 ri 220. aA 30s J. monticola 1.9 OSS 14.3)". 70. VORA 345 O07 24.2 J. flaccida 3.5 1.0 4.8. 32.0 15:9" 57.2 09 51.5 J. durangensis re US) 73” S50. 219 30% "02 7.4 J. martinezii 2.0 OS M68) Sr S40. 474 te “Sse 140 Phytologia (April 2009) 91(1) RELATIONSHIPS AND NOMENCLATURAL STATUS OF THE NOOTKA CYPRESS (CALLITROPSIS NOOTKATENSIS, CUPRESSACEAE) Zsolt Debreczy International Dendrological Research Institute P.O. Box 812910, Wellesley, MA 02482 dendro@t-online.hu, dzsgy-2@rcen.com Kathy Musial Huntington Botanical Gardens 1151 Oxford Rd., San Marino, CA 91108 Robert A. Price Biological Consulting P.O. Box 448, Alameda, CA 94501 Istvan Racz Hungarian Natural History Museum H-1087 Budapest, K6nyves K. Krt.40 ABSTRACT While working on the manuscript of Conifers Around the World (in press), the authors encountered classification and nomenclature questions surrounding the Nootka Cypress, originally described as Cupressus nootkatensis D. Don, 1824. The combination Callitropsis nootkatensis was later implicitly suggested for this taxon by Oersted as the sole species in his new genus Callitropsis, but was not published in accordance with the current International Code of Botanical Nomenclature. The combination first appears in the literature in Florin (1944), with the name attributed to Oersted, and was validated by Little (2006), who treated the species as the type of a broader genus including the New World lineage of Cupressus. The taxon has long been treated as a species of Chamaecyparis, but this placement is supported by only a limited number of non-unique morphological characters and is not supported by more recent molecular comparisons. Based on recent DNA sequence comparisons, the distinctive Nootka Cypress can appropriately be treated in a monotypic Callitropsis, in a Phytologia (April 2009) 91(1) 14] ditypic genus with the Vietnamese Yellow Cypress (originally published as Xanthocyparis vietnamensis), or in a larger generic clade with the New World Cupressus. In the following paper we discuss its complex nomenclatural and taxonomic history and morphological distinctness. Phytologia 91(1):140-159 (April, 2009). KEY WORDS: Callitropsis, Callitropsis nootkatensis, Cupressus, Xanthocyparis, Nootka Cypress, Vietnamese Yellow Cypress. BACKGROUND In 2005, while working on the manuscript of Conifers Around the World (Debreczy and Racz, in press), the authors came across classification and nomenclature questions surrounding the North American taxon well-known by its common names Nootka Cypress, Alaska Cedar, Yellow Cedar, and Alaska Yellow Cedar (Little, 1980; Rehder, 1940). In this paper we refer to it as Nootka Cypress, following its original scientific name. It was first published as Cupressus nootkatensis D. Don in Lambert, Descr. Pinus 2: 18, 1824, and has subsequently been placed in three other genera: Chamaecyparis (1841), Callitropsis (1864), and most recently, Xanthocyparis (2002). Due to its combination of vegetative and reproductive characters sharing some features with both Cupressus and Chamaecyparis, its taxonomic position has long been debated, and its nomenclature has also been subject to confusion. NOMENCLATURE OF NOOTKA CYPRESS Danish botanist Anders Sandoe Orsted (=Oersted; F ig. 1), ina detailed, richly illustrated 1864 publication, considered the cone structure of Nootka Cypress distinct enough from Chamaecyparis for the taxon to be placed in its own genus, which he named Callitropsis. Though Oersted gave a Latin diagnosis for the genus Callitropsis, and assigned only Chamaecyparis nootkatensis (“nutkaensis”) to it, he did not directly write out the new combination Callitropsis nootkatensis in accordance with the International Code of Botanical Nomenclature (ICBN) rules for pre-1953 publication of botanical names (McNeill et 142 Phytologia (April 2009) 91(1) al., 2006). Therefore the combination was not validly published and has been dealt with in a variety of ways by subsequent authors. The genus name Callitropsis Oersted and the combination Callitropsis nootkatensis were noted by (Carl) Rudolf Florin (1944) when he published the genus name Neocallitropsis as an avowed substitute for the later homonym Callitropsis Compton (Compton, 1922, p. 432). Though the name Callitropsis nootkatensis was attributed to Oersted, it was apparently first written out by Florin (1944), though not as a “comb. nov.” in accordance with the ICBN. Little (2006) cited Florin as the author of the combination (see below), apparently validating the name (Gandhi, pers. comm.). The name Callitropsis nootkatensis Oersted was also cited by Erdtman and Norin (1966) in a footnote in relation to its chemical distinctness from Chamaecyparis, but not in a nomenclatural context. The name Callitropsis nootkatensis then faded into obscurity, and the species was widely treated as Chamaecyparis nootkatensis (D. Don) Spach until evidence was obtained from tropolone and biflavonoid chemistry (Erdtman and Norin, 1966; Gadek and Quinn, 1985) and from phylogenetic analyses of morphology and DNA sequence data (Gadek et al., 2000; Farjon et al., 2002; Little et al., 2004; Xiang and Li, 2005; Little, 2006) that the species was misplaced in Chamaecyparis. Even though Florin's substitution of Neocallitropsis for Callitropsis Compton was widely accepted, the basis of that change, Oersted's name Callitropsis, was infrequently used in the literature until Little et al. (2004). When a new cupressoid conifer was discovered in karst areas of northern Vietnam in 1999, it was soon described as the new genus Xanthocyparis Farjon & H. T. Nguyén and species Xanthocyparis vietnamensis Farjon & H. T. Nguyén (Farjon et al., 2002). These authors found X. vietnamensis (Vietnamese Yellow Cypress) to be so similar in cone morphology to Nootka Cypress that they included the latter in the new genus and renamed it Xanthocyparis nootkatensis (D. Don) Farjon & Harder. However, Little et al. (2004) pointed out that if treating the two species as members of the same genus, the name Xanthocyparis was invalid since Callitropsis Oersted had priority. Phytologia (April 2009) 91 (1) 143 In February 2006 a proposal to conserve the name Xanthocyparis against Callitropsis Oersted was published (Mill and Farjon, 2006). Mill and Farjon, while also pointing out that Oersted did not make the new combination in the current manner, acknowledged that Callitropsis Oersted was validly published and that it should have been adopted for the new Vietnamese conifer and Nootka Cypress, making their publication of the name Xanthocyparis illegitimate according to ICBN Art. 52.1 (McNeill et al., 2006). Thus the present authors believe that the generic name Callitropsis Oersted should be given continued priority over Xanthocyparis when the two species are placed in the same genus, that Callitropsis Oersted is the correct generic name for the Nootka Cypress when the genus is treated as monotypic, and a monotypic Xanthocyparis is valid as its type is X. vietnamensis, not Nootka Cypress. Although in 2007 the Nomenclature Committee for Vascular Plants of the International Association for Plant Taxonomy (IAPT) voted to recommend conservation of Xanthocyparis over Callitropsis when the two species are placed in the same genus (Brummitt, 2007), the present authors believe that use of the much earlier generic name Callitropsis will cause no undue problems and that the customary rule of priority should be applied. Thus, this issue should be revisited before being voted on by the broader membership of the IAPT at the Eighteenth International Botanical Congress in 2011. In October 2006, in a paper emphasizing phylogenetic analyses of nuclear and chloroplast DNA as well as morphological data, Little (2006) retained the generic name Callitropsis, but applied it to a broader lineage including C. nootkatensis, Xanthocyparis vietnamensis, and the New World lineage of Cupressus, a taxonomic judgment that we discuss under “Generic Classification” below. In 2004 Little et al. cited the combination as “C[allitropsis]. nootkatensis (D. Don) Oerst. Apparently recognizing the problems with the publication of the combination, Little subsequently (2006) attributed Callitropsis nootkatensis to Florin. At the top of his Taxonomic Treatment, Little appears to cite the type species of Callitropsis Oersted as “Callitropsis nootkatensis (D. Don in Lambert) Florin, Regnum Veg. 100: 266. 1979.” Regnum Vegetabile 100 is the Index Nominum Genericorum (Plantarum), in which the type of the genus name is listed as Cupressus nootkatensis D. Don. The name Callitropsis nootkatensis does not 144 Phytologia (April 2009) 91(1) appear there (Farr et al, 1979). Later in his list of combinations recognized in the expanded genus Callitropsis, Little more appropriately cites the species as “Callitropsis nootkatensis (D. Don in Lambert) Florin, Palaeontographica, Abt. B, Palaéophytol. 85:590. 1944”, Florin (1944), as previously noted, did write out the combination Callitropsis nootkatensis as a name from Oersted, but did not formally propose it as a new combination, instead later referring to the taxon as Chamaecyparis nootkatensis (p. 606, l.c.). One can understandably regard Callitropsis nootkatensis (D. Don) Oersted or Callitropsis nootkatensis (D. Don) Florin as implicit combinations that were validly published under pre-1953 rules of the ICBN (vide Articles 33.2 and 33.3, McNeill, 2006). However, the combination did not appear in the International Plant Names Index (IPNI) as of 31 December 2008. While preparing this manuscript, a query from author Musial to a colleague to clarify a discrepancy in the D. Don citation eventually led to Dr. Kanchi Gandhi of the Gray Herbarium, Harvard University (also an editor for IPNI). An unexpected outcome of the correspondence on 16 January 2009 was that on 17 January 2009 “Callitropsis nootkatensis Oerst. nom. inval.” and “Callitropsis nootkatensis Oerst. ex Florin” were posted to IPNI. Further queries by Musial led Gandhi to maintain that the validity of the Florin (1944) publication was questionable and that Little (2006) might have inadvertently validated the name (Gandhi, pers. comm.). The complexity of the issue led Gandhi to consult with other IPNI editors and experts (see acknowledgements), and as of 26 January 2009, Callitropsis nootkatensis Oerst. ex Florin was also declared nom. inval. and the name validated as Callitropsis nootkatensis (D. Don) Florin ex D. P. Little, Syst. Bot. 31(3): 474, 2006; basionym Cupressus nootkatensis D. Don in Lambert, Descr. Pinus 2: 18, 1824 (IPNI, 2009). Little had formally recognized the taxon and cited its basionym in accordance with ICBN Art. 33.4, 34.1, and 46.4 (McNeill, 2006). Gandhi (pers. comm.) mentioned two alternative citations: Callitropsis nootkatensis (D. Don) Oersted ex D. P. Little or Callitropsis nootkatensis (D. Don) D. P. Little. Present authors preference is for Oersted to be credited, and on 27 January 2009 Gandhi agreed and amended the IPNI record to Callitropsis nootkatensis (D. Don) Oersted ex D. P. Little (IPNI, 2009). Phytologia (April 2009) 91(1) 145 GENERIC CLASSIFICATION OF THE NOOTKA CYPRESS Generic delimitation in the Cupressaceae sensu stricto has been more subjective than in most families of conifers due to the limited number of reproductive and vegetative characters distinguishing the approximately twenty currently recognized genera (Farjon, 2005). DNA sequence comparisons have provided independent appraisals of the relationships among these genera, and support a natural grouping including the cypresses (Cupressus sensu lato), junipers, Nootka Cypress, and Vietnamese Yellow Cypress (Gadek et al., 2000; Little et al., 2004; Xiang and Li, 2005; Little, 2006). Within this lineage, ITS sequence comparisons suggest that Nootka Cypress is the closest relative of Vietnamese Yellow Cypress (Little et al., 2004; Xiang and Li, 2005). This is consistent with the similarities in seed cone and pollen cone morphology noted by Farjon et al. (2002), but the support for a ditypic lineage comprising these two species is not strong in the other phylogenetic analyses presented by Little (2006). Thus using a total-evidence approach one can either recognize both as monotypic genera, likely with a long separate evolutionary history but with limited morphological differentiation, or treat them together as a ditypic genus. There is strong support from several lines of DNA sequence data for the inclusion of these two species in a broader phylogenetic group also including an additional well-supported lineage, the New World species of Cupressus (Little et al., 2004; Xiang and Li, 2005; Little, 2006). This has been a surprise to morphological systematists, since the New World Cupressus species are characterized by large, many-seeded, serotinous (with few exceptions) seed-cones that are retained for long periods on the shoots, and thus appear more similar to the Old World species of Cupressus than to the Nootka Cypress and Vietnamese Yellow Cypress. The molecular groupings are consistent, however, with the fact that the Nootka Cypress is crossable with several species of New World Cupressus (Jackson and Dallimore, 1926; Mitchell, 1970). The widely grown Leyland Cypress (Chamaecyparis nootkatensis x Cupressus macrocarpa) is apparently at least sometimes fertile (Jackson and Dallimore, 1926), which is highly unusual for conifers if these groups are regarded as separate genera, and also suggests that they are closely related. Thus, Little (2006) has treated the Nootka Cypress, Vietnamese Yellow Cypress, and New World 146 Phytologia (April 2009) 91(1) Cupressus in a single genus as species of Callitropsis, a rather unorthodox approach given the differences in morphology between these taxa, but fully consistent with the molecular phylogenetic groupings of the taxa without requiring a new generic name for the New World cypresses. From a macro-morphological aspect Nootka Cypress stands out from both Cupressus and Chamaecyparis and it has distinct differences from Xanthocyparis (see “Morphological Distinctness” below). Nootka Cypress can be considered as a “chamaecyparoid” cypress that, like Chamaecyparis, currently occurs in cold-temperate climates; in the case of Nootka Cypress, specifically in cool-wet boreal forests reaching as far north as 60°N. In submediterranean climates (home to regionally adjacent “true cypresses”) it is restricted to cool north slopes and high elevations where it even occurs as a groundcover shrub (Griffin & Critchfield, 1976). Occurring from extreme northwestern California to Alaska, Nootka Cypress is one of the most northern-ranging members of the Cupressaceae. The species traditionally placed in Cupressus (Old World as well as New World lineages) often occur in zonal to extrazonal mediterranean or submediterranean climates well reflected in their mostly small to medium size and upright raceme-type branchlet system. The subtropical or summer-rain tropical taxa of the genus that are adapted to humid climates and face strong competition from broad-leaved angiosperm trees are large trees with often pendulous fern-like sprays or filiferous foliage, resulting in a relatively large assimilation surface and a rain (or snow) shedding foliage system (e.g. Cupressus cashmeriana, C. funebris, C. lusitanica). MORPHOLOGICAL DISTINCTNESS OF NOOTKA CYPRESS VERSUS CHAMAECYPARIS, CUPRESSUS, AND XANTHOCYPARIS Compared with Chamaecyparis (Fig. 2). Nootka Cypress is similar to species of Chamaecyparis in having flattened branchlets, conduplicate lateral scale-leaves, and small globose cones with few basally developing seeds (2-4 per cone-scale). Nootka Cypress differs significantly from all Chamaecyparis species in its wood and leaf chemistry (Erdtman and Norin, 1966; Gadek and Quinn, 1985) and is Phytologia (April 2009) 91(1) 147 placed outside of Chamaecyparis in a separate lineage with the chemically more similar Cupressus and Juniperus in DNA sequence comparisons (Xiang and Li, 2005; Little, 2006). Morphologically, Nootka Cypress differs from Chamaecyparis in its uniform amphistomatic adult scale-leaves without obvious white stomatal patches on the down-facing sides of the branchlets, the whorl-like arrangement of its 2(—3) pairs of cone-scales, without rudimentary sterile terminal scale pairs but with a free cone-axis tip (columella), and cones maturing in (1—)2 years. Chamaecyparis has strongly dimorphic facial and lateral scale-leaves, stomata arranged in (pruinose) patches on the down-facing side of the branchlets, cones with clearly decussately developing (5)6—12 scales (3-6 pairs), with the 2-4 terminal scales sterile and connate to form a column (Jagel and Stiitzel, 2001). In its overall morphology, Nootka Cypress appears to be more of a “chamaecyparoid” (mesomorphic) Cupressus-relative than a cupressoid Chamaecyparis. The cones of Callitropsis nootkatensis (a, fig. 2, top) have a conspicuous resin-filled conical extension (columella) beyond the base of the terminal cone-scales, a feature otherwise only typical of the Australasian genus Callitris and relatives from Cupressaceae subfamily Callitroideae (inset: h, fig. 2, Callitris rhomboide, 1: vasculature of Callitris preissii). In X. vietnamensis (b, fig. 2, top) the columella is rudimentary (only a slightly raised area that can barely be considered column-like). The other genera have a longer cone-axis (relative to their cone sizes) associated with a usually larger number of cone-scales with terminal cone-scales fertile (Cupressus), or a few pairs form a sterile apical column. Compared with Cupressus (Fig. 2). Nootka Cypress is similar to the New World Cupressus species in having more or less globose seed-cones often maturing in 2 years and adult foliage that is uniform with amphistomatic scale-leaves. It differs in having relatively small seed-cones (ca. 1 cm vs. 14 cm) that open in 1-2 years rather than often being retained for long periods on the tree and opening in response to fire. The cone-scales in Nootka Cypress are basifixed and not heavily thickened, while they are medifixed (peltate) and often much thickened and woody in Cupressus. Seeds are relatively few per cone-scale (2-4), flattened, and broadly winged, versus many per cone- scale (5-20), typically lenticular or faceted, and narrowly winged in Cupressus. Pollen cones have only 2(-3) large pollen sacs per 148 Phytologia (April 2009) 91(1) sporophyll, vs. 3-6 (up to 10 in C. macrocarpa) smaller pollen sacs in Cupressus. DNA sequence studies (Xiang and Li, 2005; Little, 2006) strongly indicate that the Nootka Cypress is a close relative of the New World cypress lineage but there is no evidence that it or the related Vietnamese Yellow Cypress is derived from within the New World cypress lineage. Instead they are positioned as the closest outside relatives, as suggested by the morphological differences. Compared with Xanthocyparis vietnamensis (Figs. 3-4). The Nootka Cypress is most similar to the Vietnamese Yellow Cypress in having small more or less globose seed-cones (ca. 1 cm) with 2(—3) pairs of cone-scales on a short axis (thus appearing in whorls) with relatively few seeds per cone-scale (generally 2-4). The seeds of both are flattened and have two thin lateral wings. The pollen cones have 2(— 3) relatively large pollen sacs per sporophyll. None of these shared characters are unique in the Cupressaceae and thus they provide only limited support for a distinct phylogenetic lineage consisting of these two species (Little, 2006). The two species differ in several morphological characters with uncertain phylogenetic importance such as scale-leaf, cone, and seed properties but differ most prominently in that leaves of both the needle-like juvenile form and scale-like adult form are commonly found on adult trees of the Vietnamese Yellow Cypress, and this is not the case in the Nootka Cypress or New World cypresses. In Nootka Cypress the columella terminating the cone-axis is usually evident, while it is very reduced or rudimentary in Vietnamese Yellow Cypress (Figs. 3-4). The seeds are smooth in Nootka Cypress but are conspicuously “warty” from tiny resin-blisters in Vietnamese Yellow Cypress DNA studies of the ITS region tend to support a close sister- group relationship between these two species (Little et al., 2004; Xiang and Li, 2005), but other DNA sequence comparisons place them near one another in an unresolved trichotomy with the New World cypress lineage. The lineage including Callitropsis nootkatensis has an extensive fossil record in western North America dating back to ca. 50 MYA in the Eocene Epoch (Edwards, 1983, 1984). The chemistry of the Vietnamese Yellow Cypress is apparently not yet studied, but based on our current knowledge this would be unlikely to resolve generic relationships in the group. Phytologia (April 2009) 91(1) 149 At this time we choose to maintain Callitropsis nootkatensis and Xanthocyparis vietnamensis as members of closely related monotypic genera, which given their substantial geographic separation have probably had long evolutionary histories. Further study may provide new morphological or molecular characters that are uniquely shared by these two species or these two plus the New World cypresses, which would more strongly support a broader genus Callitropsis. SUMMARY The tortuous nomenclatural history of Callitropsis nootkatensis has apparently been resolved and the combination Callitropsis nootkatensis 1s now considered validly published and should be attributed to (D. Don) Oersted ex D. P. Little. Xanthocyparis is a valid name without need of conservation as long as the genus is kept monotypic with X. vietnamensis as its sole species. The morphological similarities between these two species, primarily in seed- and pollen-cone structure, are not unique within the family and thus may constitute only equivocal evidence in support of a separate generic lineage. Thus from a classification standpoint, the Nootka Cypress and Vietnamese Yellow Cypress are probably best considered members of closely related monotypic genera (Callitropsis nootkatensis and Xanthocyparis vietnamensis respectively) until stronger support of their phylogenetic relationship is available. ACKNOWLEDGEMENTS Authors are grateful to Judy Warnement, Director of the Harvard University Botany Libraries, for clarifying the D. Don citation in Lambert’s Description of the Genus Pinus, and to Kanchi Gandhi of the Gray Herbarium, Harvard University, and his colleagues at IPNI and other experts for resolution of the Callitropsis nootkatensis nomenclature (Werner Greuter and Eckhard Von Raab-Straube, Botanischer Garten und _ Botanisches Museum Berlin-Dahlem, Zentraleinrichtung der Freien Universitat Berlin; John McNeill, Royal Botanic Garden Edinburgh; John H. Wiersema, Systematic Botany and Mycology Laboratory, USDA/ARS, Beltsville, Maryland; John L. Strother, University of California Herbarium, Berkeley). Thanks also go to Duong Duc Huyen (Department of Botany, National Herbarium, 150 Phytologia (April 2009) 91(1) Institute of Ecology and Biological Resources of the National Center of Natural Sciences and Technology of Vietnam) for his assistance in the fieldwork to document Xanthocyparis vietnamensis, and the Botanical Library, University of Copenhagen, Denmark, for supplying Oersted’s 1864 paper and biographical information on him. Thanks to Emese Barezi and Fanni Vamos for illustrations in Figure 3. We are also grateful to Steve Edwards and Guy Nesom for reviewing the manuscript and James P. Folsom for helpful comments. LITERATURE CITED Brummitt, R. K. 2007. Report of the Nomenclatural Committee for Vascular Plants: 59. Taxon 56(4): 1289-1296. Compton, R. H. 1922. A systematic account of the plants collected in New Caledonia and the Isle of Pines. J. Linn. Soc. Bot. 45: 421- 434 + pl. Debreczy, Z., and I. Racz. 2009 (in press). Conifers Around the World. Budapest: Dendropress. Edwards, S. W. 1983. Cenozoic history of Alaskan and Port Orford Chamaecyparis cedars. Ph.D. diss., Univ. of California, Berkeley, ZI App) Edwards, S. W. 1984. New light on Alaskan and Port Orford Chamaecyparis Cedars. Four Seasons 7(2): 4-15. Erdtman, H., and T. Norin. 1966. The chemistry of the Order Cupressales. Fortschritte der Chemie Organischer Naturstoffe 24: 206-287. Farjon, A., T. H. Nguyén, D. K. Harder, K. L. Phan, and L. Averyanov. 2002. A new genus and species in Cupressaceae (Coniferales) from Northern Vietnam, Xanthocyparis vietnamensis. Novon 12: 179- 189. Farjon, A. 2005. A Monograph of Cupressaceae and Sciadopitys. London: Royal Botanic Gardens, Kew. Farr, E. R., J. A. Leissink, and F. A. Stafleu, eds. 1979. Index Nominum Genericorum (Plantarum), vol. Il. Regnum Vegetabile 100. Utrecht: Bohn, Scheltema & Holkema. Florin, R. 1938-1945. Die Koniferen des Oberkarbons und des unteren Perms. Palaeontographica 85B: 1-729. [The portion of this multiyear publication that discusses C. nootkatensis is from 1944.] Phytologia (April 2009) 91(1) 15] Gadek, P. A., and C. J. Quinn. 1985. Biflavonoids of the subfamily Cupressoideae, Cupressaceae. Phytochemistry 24: 267-272. Gadek, P. A., D. L. Alpers, M. M. Heslewood, and C. J. Quinn. 2000. Relationships within Cupressaceae sensu lato: a combined morphological and molecular approach. Am. J. Bot. 87(7): 1044— 1057. Griffin, J. R. and W. B. Critchfield. 1976. The Distribution of Forest Trees in California. Res. Pap. PSW 82/1972 (reprinted with Supplement, 1976). Berkeley: U.S.D.A. Pacific Southwest Forest and Range Experiment Station. IPNI. 2009. International Plant Names Index (list of date and place of publication of plant names). Published on the Internet http://www.ipni.org/ [accessed January 16, 2009, January 26, 2009, January 27, 2009]. Jackson, A. B., and W. Dallimore. 1926. A new hybrid conifer. Bui. Misc. Inform. 3: 113-115. Jagel, A. and T. Stiitzel. 2001. Zur Abgrenzung von Chamaecyparis Spach und Cupressus L. (Cupressaceae) und die systematische Stellung von Cupressus nootkatensis D.Don [=Chamaecyparis nootkatensis (D.Don) Spach]. Feddes Repert. 112(3-4): 179-229. Little, D. P., A. E. Schwarzbach, R. P. Adams, and C.-F. Hsieh. 2004. The circumscription and phylogenetic relationships of Callitropsis and the newly described genus Xanthocyparis (Cupressaceae). Am. J. Bot. 91(11): 1872-1881 Little, D. P. 2006. Evolution and circumscription of the true cypresses (Cupressaceae: Cupressus). Syst. Bot. 31(3): 461-480. Little, E. L. 1980. The Audubon Society Field Guide to North American Trees, Western Region. New York: Alfred Knopf. McNeill, J. et al. 2006. International Code of Botanical Nomenclature (Vienna Code). Regnum Vegetabile 146. Ruggell: A.R.G. Gantner. Mill, R. R. and A. Farjon. 2006. Proposal to Conserve the Name Xanthocyparis Against Callitropsis Oerst. (Cupressaceae). Taxon 55(1): 229-231. Mitchell, A. F. 1970. A note on two hybrid cypresses. Jour. Roy. Hort. Soc. 95: 453-454. Oersted, A. S. 1864. Bidrag til Naaletraeernes Morphologi, Videnskabelige Meddelelser fra Dansk Naturhistorisk Forening I Kjobenhavn, Ser. 2, 6: 1-36 (Contributions to the Morphology of 152 Phytologia (April 2009) 91(1) Conifers, Contributions to the Natural History Society of Copenhagen). Rehder, A. 1940. Manual of Cultivated Trees and Shrubs, 2nd ed. New York: MacMillan. Xiang, Q., and J. Li. 2005. Derivation of Xanthocyparis and Juniperus from within Cupressus: Evidence from Sequences of nrDNA Internal Transcribed Spacer Region. Harvard Pap. Bot. 9(2): 375— 382. Phytologia (April 2009) 91(1) 153 Figure 1: Anders Sandoe Orsted (1816-1872), Danish botanist, mycologist, zoologist, and marine biologist. In his long-overlooked study of the differences in cone morphology of Chamaecyparis nootkatensis and other cypresses he describes a new genus for Nootka Cypress, Callitropsis. Photograph by Johannes Peterson, Courtesy of the Botanical Library, University of Copenhagen, Denmark. 154 Phytologia (April 2009) 91(1) Figs. 2-4 Comparison of Callitropsis, Xanthocyparis, Chamaecyparis and the New and Old World Cypresses. abbreviations: ad=adaxial; ab=abaxial; sd=seed; co/=columella; trp=terminal resin pit; axrp=axillary resin pit; stcs=sterile terminal cone-scale. Figure 2. Longitudinal sections of cones and dissected cone vasculature in Callitris, Callitropsis, Xanthocyparis, Chamaecyparis, and representative New World (NW) and Old World (OW) Cupressus species. (a) Callitropsis nootkatensis, (b) Xanthocyparis vietnamensis, (c) Cupressus macnabiana (NW), (d) Cupressus macrocarpa (NW), (e) Cupressus sempervirens (OW), (f) Chamaecyparis lawsoniana, (g) Chamaecyparis obtusa var. formosana, (h) Callitris rhomboidea, note that the columella is multi-parted for this species, (i) Callitris preissii. Phytologia (April 2009) 91(1) 155 Figure 2. See caption on facing page. 156 Phytologia (April 2009) 91(1) Figure 3: Selected morphological structures of A: Callitropsis nootkatensis and B: Xanthocyparis vietnamensis. (a) Spray of mature sun branchlets with detail of individual branchlet; (b) detail of shade branchlet; (c) detail of leafy shoot, a leaf, and leaf surfaces from juvenile plant; (d) leafy shoot and detail of leaf from persistent juvenile foliage on mature tree of X. vietnamensis (not present in C. nootkatensis); (e) conelet; (f) two perpendicularly oriented views of the mature and unopened seed-cone; (g) longitudinal section of seed-cone, showing columella in Callitropsis and elevated area as rudimentary columella in Xanthocyparis and seeds; (h) seeds in lateral and facial view showing warty resin-glands on the surface of X. vietnamensis only. 137 Phytologia (April 2009) 91 (1) Figure 3. See caption on facing page. 158 Phytologia (April 2009) 91(1) Fig. 4: Selected morphological features of A: Callitropsis nootkatensis and B: Xanthocyparis vietnamensis in photographs. A: (a) juvenile branchlets from young plant; (b) semijuvenile foliage of young plant; (c) shade and (d) sun branchlets of adult plant; the same cone from (e) lateral and (f) axial views and (g) longitudinal section with columella (arrow). B: (a) juvenile branchlets from young plant; (b) juvenile-type foliage from an adult plant; (c) shade and (d) sun branchlets of adult plant; the same cone from (e) lateral and (f) axial views and (g) longitudinal section with columella area with barely visible rudimentary columella thinly filled with resin (arrow). 159 Phytologia (April 2009) 91(1) Figure 4. See caption on facing page. 160 Phytologia (April 2009) 91(1) A NEW GENUS, HESPEROCYPARIS, FOR THE CYPRESSES OF THE WESTERN HEMISPHERE (CUPRESSACEAE) Robert P. Adams Biology Department, Baylor University, Box 727, Gruver, TX, 79040 Robert_ Adams@baylor.edu Jim A. Bartel U.S. Fish and Wildlife Service, Carlsbad Fish and Wildlife Office 6010 Hidden Valley Road, Suite 101 Carlsbad, CA 92011-4213 Robert A. Price Biological Consulting P.O. Box 448, Alameda, CA 94501 ABSTRACT Phylogenetic comparisons of three nuclear DNA gene regions (nrDNA(ITS), 4-coumarate: CoA ligase, abscisic acid-insensitive 3) and a chloroplast region (petN-psbM) show that the Western Hemisphere cypresses (Hesperocyparis) is a well-supported clade quite separated from the Eastern Hemisphere cypresses (Cupressus). Based on these new data and previous data, a new genus, Hesperocyparis, 1s erected for the Western Hemisphere species previously placed in the genus Cupressus (sensu lato). Hesperocyparis is most closely related to the northwestern North American Callitropsis nootkatensis and the southeast Asian Xanthocyparis vietnamensis. Morphological characters distinguishing Hesperocyparis from CC. nootkatensis and X. vietnamensis, and from the Eastern Hemisphere cypresses (Cupressus), are presented. Phytologia 91(1):160-185 (April, 2009). KEY WORDS: Cupressus, Callitropsis, Chamaecyparis, Hesperocyparis, Xanthocyparis, Juniperus, ntDNA(ITS), 4-coumarate: CoA Ligase, Abscisic acid-insensitive 3, petN, psbM, sequences, taxonomy. Phytologia (April 2009) 91(1) 16] Attempts to identify and delimit coniferous genera have been “based on limited sets of usually selective characters which were perceived to be informative about evolution and/or phylogeny of the group ... under study a priori” (Farjon, 2005). Despite the numerous taxonomic works addressing the Cupressaceae (sensu lato), which were described in detail by Farjon (2005), the modern concept of Cupressus has remained largely unchanged for more than a century. However, the discovery of a new conifer species on karst limestone in northern Vietnam (Averyanov et al., 2002; Farjon et al., 2002) has led to both excitement and taxonomic difficulties. Farjon et al. (2002) recognized the taxon as a new species and genus, Xanthocyparis vietnamensis Farjon & T. H. Nguyén, based on dimorphic leaves; small ovulate cones with 2 or 3 pairs of opposite decussate cone scales; 2 years for seed cone maturation; flattened, winged seeds; and juvenile, transition, and adult leaves found on the same tree. Farjon et al. (2002) concluded that Chamaecyparis nootkatensis (D. Don) Spach was congeneric with X. vietnamensis and included the former species in the new genus (overlooking the earlier generic name Callitropsis Oersted) and made the new combination Xanthocyparis nootkatensis (D. Don) Farjon & D. K. Harder. Chamaecyparis nootkatensis has had a variable taxonomic history, having been classified as Chamaecyparis, Cupressus, Callitropsis and Xanthocyparis (see Little et al., 2004, and Debreczy et al., 2009 for discussion). Little et al. (2004), using nrDNA(ITS) internal transcribed spacer (ITS) sequence data, found that Xanthocyparis vietnamensis and X. nootkatensis form a clade sister to the Western Hemisphere cypresses and that the Eastern Hemisphere cypresses and Juniperus constitute distinct clades outside this group. Little et al. (2004) reported that Chamaecyparis nootkatensis had been previously described as Callitropsis nootkatensis (D. Don) Oersted in 1865. Little renamed _X. vietnamensis as Callitropsis vietnamensis (Farjon and Nguyén) D. P. Little. Though Silba (2005) did not address the molecular phylogeny results, he did assert in response to Little et al. (2004) that the splitting of western and Eastern Hemisphere cypresses was “based on superficial data with inaccurate and incomplete field observations.” More recently, Mill and Farjon (2006) made a proposal to conserve 162 Phytologia (April 2009) 91(1) Xanthocyparis against Callitropsis. The Nomenclature Committee for Vascular Plants voted 14-4 to recommend that the proposal be adopted at the next International Association of Plant Taxonomists congress in 2012 (Brummitt, 2007). So, the matter currently remains unsettled (see Debreczy et al., 2009, for discussion). Xiang and Li (2005) reexamined Xanthocyparis, Chamaecyparis, Cupressus and Juniperus using nrDNA(ITS) sequences. Though the authors concluded that “it seems appropriate” to place Xanthocyparis vietnamensis and X. nootkatensis in Cupressus (sensu lato), they also noted that “Assuming the ITS tree reflects species relationships, we need a new genus name for the New World species of Cupressus if Xanthocyparis 1s recognized.” While Xiang and Li (2005) submerged X. vietnamensis in Cupressus, the combination Cupressus vietnamensis was made previously by Silba (2005) and remade later by Rushforth (2007). Prior to the recent spate of publications described above, taxonomic work on Cupressus largely was focused on the specific and infraspecific level with considerable disagreement as to the number of distinct species to recognize in the genus (see Wolf, 1948; Little, 1970; Farjon, 2005). The classical monograph by Camus (1914) treated the known species of Cupressus on a worldwide basis and also included the distinct but related Chamaecyparis as a subgenus of Cupressus. The most thorough morphological treatment of the Western Hemisphere species of Cupressus is the revision of Wolf (1948), who only included the New World species in his study on the grounds that the Eurasian and African species were not readily accessible for detailed population- based field studies. He also stated that “none of [the Old World species] appears closely related to our New World species.” Silba (1983), after reportedly raising seedlings of the 25 taxa he delimited, noted consistent differences in cotyledon number and shape in all Western versus Eastern Hemisphere cypresses (cotyledons 3-4 and acute, versus 2 and obtuse), though he did not think these characters, alone, warranted dividing Cupressus into subsections. However, Silba (1994, 1998) later arranged Cupressus into two subgenera and seven sections, and designated Cupressus lusitanica as the type for his new subgenus, /ndoamericana, which included Western Phytologia (April 2009) 91(1) 163 Hemisphere taxa of Cupressus with some additional Asian species. The name of this subgenus, /mdoamericana, reflects the generally discredited belief, which Silba (2006) continues to support, that C. lusitanica (which occurs in the wild in Mexico and central America) actually originated in Goa, India, from where seed was purportedly collected and introduced into Portugal (Farjon 1993). Apart from this problem, Silba’s (1994, 1998) treatment would place some species in multiple sections (Little, 2006). Little (2006) expanded the scope and depth of his previous work (Little, et al., 2004) and analyzed cpDNA (matkK, rbcL, and trnL) plus two nuclear gene regions: nrDNA(ITS), and NEEDLY for all 16 species of the Western Hemisphere and 12 species of the Eastern Hemisphere (Cupressus, sensu lato). The portions of his trees relating to Xanthocyparis, Cupressus, and Juniperus are depicted in figure 1. imatK, roel, tnt '! nrDNA (ITS) 99 Juniperus 7 Eastern Hemisphere cypresses Eastern Hemisphere cypresses : Juniperus X. vietnamensis C. nootkatensis X. vietnamensis C. nootkatensis i 90 Western Hemisphere piginicie "Combined ‘Molecular 4 + “Morphology _ , X. vietnamensis + 700 Western Hemisphere CYPIESSOS: C. nootkatensis i __3°X. vietnamensis Western Hemisphere cypresses || --C. nootkatensis 700 Juniperus Figure 1. Summary of partial trees extracted from data of Little (2006). Numbers below the branches are strict consensus jackknife frequencies above 50%. Dashed lines in the Combined Molecular + Morphology tree for C. nootkatensis and X. vietnamensis indicate branches that are collapsed in the strict consensus. 164 Phytologia (April 2009) 91(1) Little (2006) obtained strong support for the Western Hemisphere cypresses as a monophyletic clade in the analyses of each of his molecular data sets. In addition, there was strong support (Fig. 1) that the closest relatives of Western Hemisphere cypresses are Xanthocyparis vietnamensis and Callitropsis nootkatensis. The Western and Eastern Hemisphere cypresses, and Juniperus are each well resolved as distinct clades in each of his molecular analyses, while X. vietnamensis and C. nootkatensis form an unresolved trichotomy with the Western Hemisphere cypresses in the cpDNA and NEEDLY analyses, and are only moderately well supported (61%) as a 2-species clade in the ITS analysis (Fig. 1). A tree based on morphological data failed to separate eastern from Western Hemisphere cypresses, but a combined analysis of the morphological and molecular data sets did strongly separate these geographic groups (Little, 2006, Fig. 1 above). The results of the molecular and combined analyses also provided strong evidence that C. /usitanica is nested well within the Western Hemisphere group, whereas C. torulosa is definitely placed within the Eastern Hemisphere lineage contrary to the subgeneric classification of Silba (1994). In addition to the previously mentioned response to Little et al. (2004) by Mill and Farjon (2006), Farjon (2007) in a letter to a Taxon took issue with Little’s 2006 paper because Farjon maintained that the only significant morphological difference between the Western and Eastern Hemisphere cypresses, cotyledon number, did not hold for two Asian species, C. chengiana and torulosa. Farjon (2007) concluded that there “are no morphological or anatomical differences that justify this generic separation.” While acknowledging that “No single characteristic can be used diagnostically,” Little (2006) stated that a suite or “series of vegetative characteristics possibly associated with adaption to arid environments (e.g., monomorphic leaves, penultimate and ultimate segments arranged on two planes) unite the New World Cupressus species to the exclusion of Old World Cupressus, Juniperus, and Callitropsis [sensu stricto].” Little (2006) decided to include the Western Hemisphere cypresses (Cupressus) in Callitropsis and published 17 new names from North America. Little (2006) reasoned that giving a new genus name to the Western Hemisphere cypresses (as suggested by Xiang and Phytologia (April 2009) 91(1) 165 Li, 2005) would be “consistent with some but not all resolutions of the polytomy between Callitropsis [i.e., X. vietnamensis and Ch. nootkatensis| and the New World Cupressus species.” However, only the ITS data (Fig. 1) present X. vietnamensis - C. nootkatensis as a clade and Little’s results did not provide any strong evidence against placing these two taxa in a separate genus (as suggested by Farjon et al., 2002 and Little et al., 2004) or as monotypic genera as suggested by Debreczy et al. (2009). In an effort to add additional molecular data to the taxonomic questions, we have sequenced two nuclear genes (4-coumarate: CoA ligase, 4CL and abscisic acid-insensitive 3, ABI3, as well as complete nrDNA(ITS) sequences for additional taxa and a cpDNA region, petN- psbM. The 4-coumarate: CoA Ligase (4CL) gene family is important in phenylpropanoid synthesis leading to lignin, as well as flavonoids, and other pigments as well as phenolic compounds in essential oils such as safrole, eugenol, etc. (Hamberger and Hahlbrock, 2004; Cukovic et al., 2001) Recently, Peng and Wang (2008) utilized 4CL sequences to study Thuja species and Thujopsis dolabrata. In Thujopsis dolabrata they found the 4CL gene to be composed of 4 exons and 3 introns. Intron 2 was reported as 640 bp (EU183423). Aligning the GenBank sequences for Thuja plicata (EU183418, EU183417) and Thujopsis dolabrata (EU183423) enabled us to design primers to span intron 2, and resulted in 746 - 823 bp of sequence data. Lazarova, Zeng and Kermode (2001) reported on the occurrence of an abscisic acid-insensitive 3 (ABI3) gene homologue from Chamaecyparis nootkatensis (CnABI3). The ABI3 gene is composed of six exons and five introns, with the intron sizes of 105, 113, 110, approx. 1000, and 142 bp. Primers were designed in exon 4 and exon 5 to amplify intron 4 (see Materials and Methods below) and resulted in 1020 - 1108 bp of sequence data. The cp region trnC-trnD has been used in phylogenetic studies in Juniperus (Adams, 2007; Adams et al., 2007). The partial sequence utilized in this study is the petN - psbM region (included in the trnC- tmD region). This region is much easier to amplify and resulted in 166 Phytologia (April 2009) 91(1) approximately 807-854 bp compared to 1400 - 1500 for the full trnC- trnD region. The purpose of the present study is to bring additional molecular data to bear on the question of the taxonomic status of Xanthocyparis, versus the Eastern and Western Hemisphere cypresses. MATERIALS AND METHODS Specimens used in this study: Species Voucher Source GenBank X. vietnamensis nrDNA(ITS) Little et al. 2004 Vietnam AY 380877 ACE Rushforth 7745 Vietnam FJ744493 CnABI3 Rushforth 7745 Vietnam FJ56803 petN-psbM — Rushforth 7745 Vietnam FJ46729 C. nootkatensis nrDNA(ITS) Little et al. 2004 AK, USA AY380858 4CL Adams 9086 WA, USA FJ744494 CnABI3 Adams 9086 WA, USA FJ56803 petN-psbM Adams 9086 WA, USA FJ46730 C. atlantica nrDNA(ITS) | Little et al. 2004 Morocco AY 988367 4CL Adams 8429 Morocco FJ744495 CnABI3 Adams 8429 Morocco FJ56805 petN-psbM Adams 8429 Morocco FJ46731 C. dupreziana, nrDNA(ITS) Little et al. 2004 Algeria ex Hillier Gard. AY988375 4CL Adams 8432 Algeria ex Hillier Gard. FJ744496 CnABI3 Adams 8432 Algeria ex Hillier Gard. FJ56806 petN-psbM Adams 8432 Algeria ex Hillier Gard. FJ46733 C. sempervirens, nrDNA(ITS) Adams 8434 Elburz Mts., Iran FJ705221 4CL Adams 8434 Elburz Mts., Iran FJ744497 CnABI3 Adams 8434 Elburz Mts., Iran FJ56807 petN-psbM Adams 8434 Elburz Mts., Iran FJ46732 Phytologia (April 2009) 91(1) H. abramsiana 167 nrDNA(ITS) Adams 9354 CA, USA FJ705220 4CL Adams 9354 CA, USA FJ744498 CnABI3 Adams 9354 CA, USA FJ56808 petN-psbM Adams 9354 CA, USA FJ46737 H. bakeri nrDNA(ITS) Little et al. 2004 CA, USA AY988369 4CL Adams 9362 CA, USA FJ744499 CnABI3 Adams 9362 CA, USA FJ56809 petN-psbM Adams 9362 CA, USA FJ46739 H. pygmaea ntDNA(ITS) Adams 9357 CA, USA FJ705219 4CL Adams 9357 CA, USA FJ744500 CnABI3 Adams 9357 CA, USA FJ56810 petN-psbM Adams 9357 CA, USA FJ46738 J. monticola nrDNA(ITS) Adams 6876 HID, MX FJ705218 4CL Adams 6876 HID, MX FJ744501 CnABI3 Adams 6876 HID, MX FJ56811 petN-psbM Adams 6876 HID, MX FJ46736 J. saltillensis nrDNA(ITS) Adams 6886 NL, MX FJ705217 4CL Adams 6886 NL, MX FJ744502 CnABI3 Adams 6886 NL, MX FJ56812 petN-psbM Adams 6886 NL, MX FJ46735 J. virginiana nrDNA(ITS) Adams 6753 TX, USA EF608980 4CL Adams 6753 TX, USA FJ744503 CnABI3 Adams 6753 TX, USA FJ56813 petN-psbM Adams 6753 TX, USA FJ46734 Thujopsis dolabrata nrDNA(ITS) Peng and Wang Jiangxi, China EU183443 4CL Peng and Wang Jiangxi, China EU183423 CnABI3 Adams 9502 Japan ex Arn. Arb. FJ56814 petN-psbM Adams 9502 Japan ex Arn. Arb. FJ46727 168 Phytologia (April 2009) 91(1) Thuja plicata nrDNA(ITS) Adams 9277 Vancouver Isl., BC AY380852 4CL Peng and Wang Kew Bot. G.ex USA? EU183417 CnABI3 Adams 10311 Queen Charlotte Isl., BC FJ56815 petN-psbM Adams 10311 Queen Charlotte Isl., BC FJ46728 Specimens only used for size determination of 4CL: C. arizonica, Adams 9378, Pima Co., AZ; C. benthamii, Adams 8710, Pachuca, MX; C. forbesii, Adams 9370, San Diego Co., CA; C. glabra Adams 9389, Gila Co., AZ; C. goveniana, Adams 11544, Monterey Co., CA; C. guadalupensis, Adams 8417, Guadalupe Isl., MX, ex Berkeley Bot. Garden; C. /usitanica, Adams 7071, cultivated, Bussaco, Portugal; C. macnabiana, Adams 9359, Napa Co., CA; C. macrocarpa, Adams 11459, Crocker Grove, CA; C. montana, Adams 9660, Baja, MX; C. nevadensis, Adams 9367, Kern Co., CA; C. sargentii, Adams 9348, San Luis Obispo Co., CA; C. stephensonii, Adams 9376, San Diego Co., CA. Voucher specimens for Adams collections are deposited at BAYLU. Bartel specimens are held in his personal herbarium. One gram (fresh weight) of the foliage was placed in 20 g of activated silica gel and transported to the lab, thence stored at -20° C until the DNA was extracted. DNA was extracted from juniper leaves by use of a Qiagen mini-plant kit as per manufacturer's instructions. Amplification and sequencing ITS (nrDNA), 4CL and tmC-trnD amplifications were performed in 30 pl reactions using 6 ng of genomic DNA, 1.5 units Epi-Centre Fail-Safe Taq polymerase, 15 ul 2x buffer E or K (final concentration: 50 mM KCl, 50 mM Tris-HCl (pH 8.3), 200 uM each dNTP, plus Epi-Centre proprietary enhancers with 1.5 - 3.5 mM MgCl, according to the buffer used) 1.8 uM each primer. Gene __ Primers 2x buffer annealing program size bp nrITS ITSA/ITSB K 50°C (94-50x30) 1077-1105 4CL 4CL49F/4CL814R G 55°C (94-55x30) 746-823 CnABI3, CnABII1F/357R D S55C = (94-55-x30) 1020-1108 petN _petN5F/psbM111R_E 50°C __(94-50x30) 807-854 Primers (5'-3'): ITS: ITSA = GGA AGG AGA AGT CGT AAC AAG G; Phytologia (April 2009) 91(1) 169 Mss = CTT TTC CTC CGC ITA TTG ATA TG, ITSA and ITSB primers from Blattner (1999). 4CL:4CL49F AAAGAGCTCATCAAATACAA 4CL814R GAAGAGCTTCCAGCTCAG 4CL primers are from conserved sequences in exon 2 and exon 3 of Thuja plicata (EU183418, EU183417) and Thujopsis dolabrata (EU 1834232) and span intron 2. CnABI3: CnABI11F AACAATAAGAGCAGGATGTA CnABI357R CCAGTTTTGGTATCAGAGTA Addition internal primers utilized: CnABhint533R CAATATTATCACGCATTTG CnABhint541R CACAGGAGCAATATTATCAC CnABhint741R TTACTTGAAACAATCTATTTATGT CnABI3 primers are from sequences in exon 4 and exon 5 of Chamaecyparis nootkatensis (AJ131113) and span intron 4. petN - psbM: petNSF: AAC GAA GCG AAA ATC AAT CA psbM111R: AAA GAG AGG GAT TCG TAT GGA petN and psbM primers were based on conserved sequences from Juniperus species. The following PCR conditions were used: MJ Research Programmable Thermal Cycler, 30 cycles, 94°C (1 min.), 50°C or 57°C (2 min.), 72°C (2 min.), with a final step of 72°C (5 min.). The PCR reaction was subjected to purification by agarose gel electrophoresis (1.5% agarose, 70 v, 55 min.). In each case, the band was excised and purified using a Qiagen QIAquick gel extraction kit. The gel purified DNA band with the appropriate primer was sent to McLab Inc. for sequencing. Sequences for both strands were edited and a consensus sequence was produced using Chromas, version 2.31 (Technelysium Piy . Ltd.). Alignments were made using MAFFT (http://align.bmr.kyushu-u.ac.jp/mafft/) and then manually corrected and then re-analyzed using NJ with 1000 bootstrap replications (http://align.bmr.kyushu-u.ac.jp/mafft/). 170 Phytologia (April 2009) 91(1) We included Thuja and Thujopsis as outgroup taxa in the analyses following the phylogenies of Gadek et al. (2000) and Little et al. (2004). RESULTS AND DISCUSSION The overall sequencing efforts are shown in table 1. The number of informative sites and the percent yield varied from largest in nrDNA(ITS) to smallest in petN-psbM. Clearly, nrDNA(ITS) yielded both the most informative sites and the greatest yield for the effort. The single (or low) copy nuclear genes yielded lots of information, being single genes, are difficult to amplify in amounts for preparative yields. The cp DNA (petN-psbM) is multiple copy and very easy to amplify, but the number and yield of informative sites 1s somewhat smaller. Table 1. Summary of sequencing results. # of variable and # of informative sites are within the in-group (excluding T. dolabrata and Th. plicata). % yield of informative sites (% yield) = 100 x # informative / minimum range observed. gene range, bp _# variable _# informative % yield nrDNA(UITS) 1077-1105 198 158 14.7% 4CL 746-823 124 79 10.6 CnABI3 1020-1108 137 83 8.1 petN-psbM 807-854 84 57 Dell Sequencing the nrDNA (ITS region) resulted in 1077 to 1105 bp of sequence data. The ITS tree (Fig. 2) is similar to that of Little (2006, Fig. 1, upper right, above), in that the cypresses from the Eastern and Western Hemispheres are 100% supported as distinct clades. There is some support (75%) for the clade of C. nootkatensis - X. vietnamensis as reported by Little (2006, 61%, Fig. 1 above). The C. nootkatensis - X. vietnamensis, clade is allied with the Western Hemisphere cypresses using the ITS data (Fig. 2). Sequencing of the 4-coumarate: CoA ligase intron 2 (4CL) region resulted 746 - 823 bp of sequence data. Examination of the NJ tree reveals four groups (fig. 3) as found with the nrDNA(ITS) data (Fig. 2). The 4CL tree shows a weak association (34%) between C. Phytologia (April 2009) 91(1) 171 pygmaea nrDNA(ITS) 158 inf. nuc. abramsiana 1077-1105 bp NJ phylogram bakeri C. nootkatensis X. vietnamensis J. saltillensis 100 J. monosperma J. virginiana C. dupreziana 92 C. atlantica C. sempervirens T. dolabrata Th. plicata Figure 2. NJ phylogram based on nrDNA(ITS). Numbers below branches are bootstrap probabilities (1000 reps). Eastern Hemisphere cypresses are in the cross-hatched box and Western Hemisphere cypresses are in the shaded box. L72 Phytologia (April 2009) 91(1) C. atlantica 4CL 4-coumarate: CoA Ligase 43 79 inf. nuc. C. sempervirens 746-823 bp C. dupreziana NJ phylogram C. nootkatensis X. vietnamensis J. saltillensis 98 J. monticola J. virginiana pygmaea abramsiana bakeri T. dolabrata Th. plicata Figure 3. NJ phylogram utilizing sequences from intron 2 of 4- coumarate: CoA ligase (4CL). Phytologia (April 2009) 91(1) i723 nootkatensis-X. vietnamensis and the Eastern Hemisphere cypresses, but again provides substantial support (99%) for the Western Hemisphere cypress clade. The three Western Hemisphere cypresses (C. abramsiana, C. bakeri and C. pygmaea) each had a unique 46 bp insert giving them a 4CL length of 817 bp, in contrast, all other taxa that had shorter sequences. A survey of all the other Western Hemisphere cypresses (C. arizonica, C. benthamii, C. forbesii, C. glabra, C. goveniana, C. guadalupensis, C. lusitanica, C. macnabiana, C. macrocarpa, C. montana, C. nevadensis, C. sargentii, C. stephensonii) revealed that the length is nearly constant at 817bp, indicating that all these taxa share the 46bp insert. Sequencing of the CnABI3 intron 4 region revealed several large indels in this data set. The NJ phylogram based on CnABI3 sequence data again shows (Fig. 4) the separate clades of the Eastern and Western Hemisphere cypresses. However, C. nootkatensis and X. vietnamensis do not form a clade but are well supported as species. The CnABI3 gene sequence supports the contention of Debreczy et al. (2009) that C. nootkatensis and X. vietnamensis are monotypic genera, since the former forms a strongly supported clade with the Western Hemisphere cypresses (99%). It is interesting to note that X. vietnamensis, C. atlantica, C. dupreziana and C. sempervirens all share a unique 47 bp deletion. Sequencing petN-psbM of cpDNA resulted lengths ranged from 807 to 854 bp, except for 7. dolabrata that had only 511 bp. The NJ phylogram (Fig. 5) again shows strong support for separate clades for the Eastern and Western Hemisphere cypresses. Overall, the tree is similar to the cpDNA tree of Little (2006) based on combined sequences from matK, rbcL and trnL (Fig. 1, upper left), but the greater amount of sequence data in Little’s tree provides very strong support for the monophyly of the Western Hemisphere cypress lineage (100%), while C. nootkatensis and X. vietnamensis are not resolved from the Western Hemisphere cypresses in our analysis (Fig. 5). However, again, the Eastern and Western Hemisphere cypresses are in well- supported clades (Fig. 5). 174 ABI3 abscisic acid-insensitive 3 83 inf. nuc. 1020-1108 bp NJ phylogram Phytologia (April 2009) 91(1) pygmaea bakeri abramsiana C. nootkatensis X. vietnamensis J. saltillensis J. monticola J. virginiana C. atlantica 99 C. sempervirens C. dupreziana T. dolabrata Th. plicata Figure 4. NJ phylogram based on ABI3 intron 4 sequences. Phytologia (April 2009) 91(1) 175 petN-psbM abramsiana 57 inf. nuc. 807-854 bp PYSBCS NJ phylogram bakeri C. nootkatensis X. vietnamensis C. atlantica C. dupreziana C. sempervirens J. saltillensis J. monticola J. virginiana T. dolabrata Th. plicata Figure 5. NJ phylogram based on petN - psbM sequences of cp DNA. 176 Phytologia (April 2009) 91(1) An analysis based on combined sequences (nrDNA(ITS), 4CL, CnABI3, petN-psbM) yielding 377 phylogenetically informative nucleotides. The NJ phylogram (Fig. 6) has 100% support for the 7. dolabrata - Th. plicata, Western Hemisphere cypresses, Eastern Hemisphere cypresses, and Juniperus clades. It also provides 100% support for the grouping of C. nootkatensis and X. vietnamensis with the Western Hemisphere cypresses, and is consistent with the proposal by Debreczy et al. (2009) to treat these taxa as monotypic genera. The separation of C. nootkatensis and X. vietnamensis is consistent with several morphological characters distinguishing the two: both needle-like juvenile leaves and scale-like adult leaves occur on the mature plant in only the latter, the seed coat has minute warty resin pustules in only the latter, and there is a short but quite distinct resin-filled columella at the center of the mature and open seed cone in only the former (Debreczy et al., 2009). Callitropsis nootkatensis and similar extinct forms also have a substantial fossil record dating back to at least 50 MYA in western North America (Edwards, 1983), which may serve to provide a minimum time depth for the split between this group and the Western Hemisphere cypress lineage. Our results are consistent with those of Little (2006) in providing further support for a distinct lineage of Western Hemisphere cypresses quite separate from the Eastern Hemisphere cypresses and most closely related to C. nootkatensis and X. vietnamensis. The Western Hemisphere cypresses differ significantly in cone morphology from the latter two species, notably in having cones with many more seeds (typically 5-20 per cone scale and 60-150 per cone versus 2-4 per cone scale and < 15 per cone), woodier and larger peltate cone scales, and cotyledons, with few exceptions, 3-5 in number versus 2. Thus, rather than following Little (2006), who included the Western Hemisphere cypresses plus C. nootkatensis and X. vietnamensis in an expanded genus Callitropsis, we recognize a new genus including only the Western Hemisphere cypress lineage. The traditional approach of including the Eastern Hemisphere and Western Hemisphere cypress lineages in a genus to the exclusion of Phytologia (April 2009) 91(1) 177 ar Combined Molecular ike ints 377 inf. nuc. 3652-3883 bp pygmaea NJ phylogram baker C. nootkatensis X. vietnamensis J. monticola J. saltillensis J. virginiana C. atlantica C. semipervirens C. dupreziana T. dolabrata Th. plicata Figure 6. NJ phylogram based on combined sequences (nrDNA(ITS), 4CL, CnABI3, petN-psbM). 178 Phytologia (April 2009) 91(1) Callitropsis, Xanthocyparis and Juniperus would run contrary to the very likely phylogenetic relationships in the group supported by multiple lines of molecular phylogenetic data, which indicate that C. nootkatensis and X. vietnamensis rather than the Eastern Hemisphere cypresses are the closest relatives of the Western Hemisphere cypresses (Figs. 1, 6). We believe that the possible alternative approach of including the entire clade of cypresses, junipers, Xanthocyparis and Callitropsis in a single genus would be unduly disruptive to the nomenclature of horticulturally important taxa, particularly if Cupressus is given nomenclatural priority over Juniperus (which would require 67 new combinations at the species level; Adams, 2008), and would also tend to obscure rather than elucidate the morphological groupings and major evolutionary lineages in the group. We provisionally recognize 16 species as distinct for purposes of providing new species combinations, following the monographic treatment of Wolf (1948) and the phylogenetic results of Little (2006). The new genus is cryptic in its macromorphology, being similar to Cupressus stricto sensu in its general appearance and cone morphology, but is very distinct in molecular phylogenetic analyses from multiple genes and two genomes. In morphology, it is most evidently distinguished from the majority of species of Cupressus stricto sensu in its greater number of cotyledons (3-5), and is distinguished from any taxa of Eastern Hemisphere cypresses that may have parallelisms for this character (C. torulosa of Asia has 3-5 cotyledons according to Camus, 1914) by a combination of branchlet characters as described below. Cupressus torulosa and all other native Eurasian and African species of the genus are unequivocally placed in the Eastern Hemisphere clade in the molecular phylogenetic analyses of Little (2006). Recognition of new genera as new sources of phylogenetic information emerge to support them as _ distinct evolutionary units has a long tradition, as witness the segregation of multiple genera of Cupressaceae with similar cone morphology from the classical genus Libocedrus. These segregate genera are now widely recognized (Farjon, 1998, 2005) and are well supported by recent molecular studies (Gadek et al., 2000). Phytologia (April 2009) 91(1) 179 TAXONOMIC TREATMENT Hesperocyparis Bartel & R. A. Price, gen. nov.— TYPE: Hesperocyparis macrocarpa (Hartw. ex Gordon) Bartel. Differt a Callitropse and Xanthocypare cotyledonibus 3-—S (vs. 2), squamis strobilis paribus 3—6 (vs. 2—3) peltatis non dense incrassatis (vs. basifixis non dense incrassatis), et seminibus per strobilum generaliter 60-150 (vs. paucioribus quam 15). Differt a Cupresso cotyledonibus 3—5 (vs. plerumque 2), testa generaliter glauco (vs. non glauco), ordinibus ultimis duobus segmentis caulinis in fasciculis 3- dimensionalibus, segmentis ultimis caulibus in sectione transversali non complanatis, et foliis monomorphis segmentorum caulinorum ultimorum (vs. ordinibus ultimis duobus segmentis caulinis in asperginibus 2-dimensionalibus aut segmentis caulinis ultimis in sectione transversali complanatis et foliis dimorphis segmentorum caulinorum ultimorum). Plantae Hemisphaerii Occidentalis. Hesperocyparis differs from Callitropsis and Xanthocyparis in its cotyledons 3-5 (vs 2), seed cone scales in 3-6 pairs (vs 2-3 pairs), peltate and heavily thickened (vs basifixed and not heavily thickened), and seeds per cone generally 60-150 (vs < 15). Hesperocyparis differs from Cupressus in its cotyledons 3-5 (vs usually 2), seed coat generally + glaucous (vs not glaucous), usually ultimate 2 orders of branch segments in 3-dimensional clusters, ultimate branch segments not flattened in cross section, and ultimate branch segments leaves monomorphic (vs usually ultimate 2 orders of branch segments in 2- dimensional sprays, or ultimate branch segments flattened in cross section and ultimate branch segments leaves dimorphic). Plants of the Western Hemisphere. Shrub or tree to (<1-)4-35(-40) m, multi- to generally single- trunked, monoecious, evergreen. Bark on trunk fibrous or leathery and smooth, exfoliating in fibrous strips or irregular-shaped plates, gray to brown to cherry-brown. Branch segments (stems and overlapping leaves) terete to quadrangular, ultimate and penultimate branch segments generally in 3-dimensional clusters or rarely in 2-dimensional flattened sprays. Leaves of juvenile plants awl- to needle-like, 180 Phytologia (April 2009) 91(1) decussate or in whorls of 3; of adult plants decussate, scale-like, appressed, overlapping, generally monomorphic, minutely denticulate or rarely entire, often with a dorsal resin gland, leaves on vigorously growing shoots more elongate and acute-tipped. Pollen cones, terminal on separate ultimate branch segments, sub-spheric to elliptic-ovoid to cylindrical, terete to quadrangular, 2.0-6.5 mm long, 1.3-3.0 mm wide, yellow-green; microsporophylls decussate in 3-10 pairs, 3-6(10) sporangia in an irregular row per microsporophyll. Seed cones 10-50 mm long, more or less woody, nearly spheric to widely cylindric, maturing in the second year, generally remaining closed at maturity and opening after many years or in response to fire, abscising after opening or after many additional years; scales decussate in (2-)3-6 pairs, thickened, peltate, abutting, shield- or wedge-shaped, boss generally >1 mm (especially prior to maturity), pointed, base level with or rising from edge. Seeds many per scale (generally 5-20) per cone, flattened, ovate to lenticular, irregularly faceted due to close packing; seed wings, 2, membranous, narrow, seed body light tan to red brown to brown to dark brown to black, generally glaucous, generally warty with minute resin pustules in the seed coat; cotyledons (2-)3-5(-6), linear, slightly ridged, bluntly pointed at apex. Chromosome number, 2n = 22(23,24). Hesperocyparis abramsiana (C. B. Wolf) Bartel, comb. nov. Cupressus abramsiana C. B. Wolf, Aliso 1: 215. 1948. Cupressus goveniana Gordon var. abramsiana (C. B. Wolf) Little, Phytologia 20: 435. 1970. Cupressus goveniana Gordon subsp. abramsiana (C. B. Wolf) A. E. Murray, Kalmia 12: 19. 1982. Callitropsis abramsiana (C. B. Wolf) D. P. Little, Syst. Bot. 31: 473. 2006. Hesperocyparis arizonica (Greene) Bartel, comb. nov. Cupressus arizonica Greene, Bull. Torrey Bot. Club 9: 64. 1882. Cupressus benthamii Endl. var. arizonica (Greene) Mast., J. Linn. Soc. Bot. 31: 340. 1896. Callitropsis arizonica (Greene) D. P. Little, Syst. Bot. 31: 473. 2006. Hesperocyparis bakeri (Jeps.) Bartel, comb. noy. Cupressus bakeri Jeps., Fl. Calif. 1: 61. 1909. Cupressus macnabiana A. Murray bis var. bakeri (Jeps.) Jeps., Man. Fl. Pl. Calif. 58. Phytologia (April 2009) 91(1) 181 1923. Callitropsis bakeri (Jeps.) D. P. Little, Syst. Bot. 31: 473. 2006. Hesperocyparis benthamii (Endl.) Bartel, comb. nov. Cupressus benthamii Endl., Syn. Conif. 59. 1847. Cupressus lusitanica Mill. var. benthamii (Endl.) Carriere, Traité Gén. Conif., ed. 2, 155. 1867. Cupressus lusitanica Mill. subsp. benthamii (Endl.) Franco, Agros (Lisbon) 28: 24. 1945. Callitropsis benthamii (Endl.) D. P. Little, Syst. Bot. 31: 473. 2006. Hesperocyparis forbesii (Jeps.) Bartel, comb. nov. Cupressus forbesii Jeps., Madrofio 1: 75. 1922. Cupressus guadalupensis S. Watson var. forbesii (Jeps.) Little, Phytologia 20: 435. 1970. Cupressus guadalupensis S. Watson subsp. forbesii (Jeps.) R. M. Beauch., Aliso 9: 191. 1978. Callitropsis forbesii (Jeps.) D. P. Little, Syst. Bot. 31: 473. 2006. Hesperocyparis glabra (Sudw.) Bartel, comb. nov. Cupressus glabra Sudw., Amer. Forest. 16: 88. 1910. Cupressus arizonica Greene var. glabra (Sudw.) Little, Madrofio 18: 162. 1966. Cupressus arizonica Greene subsp. glabra (Sudw.) A. E. Murray, Kalmia 12: 19. 1982. Callitropsis glabra (Sudw.) D. P. Little, Syst. Bot. 31: 473. 2006. Hesperocyparis goveniana (Gordon) Bartel, comb. nov. Cupressus goveniana Gordon, J. Hort. Soc. London 4: 295. 1849. Callitropsis goveniana (Gordon) D. P. Little, Syst. Bot. 31: 473. 2006. Hesperocyparis guadalupensis (S. Watson) Bartel, comb. nov. Cupressus guadalupensis S. Watson, Proc. Amer. Acad. Arts 14: 300. 1879. Cupressus macrocarpa_ Hartw. var. guadalupensis (S. Watson) Mast., Gard. Chron., Ser. 3 18: 62. 1895. Callitropsis guadalupensis (S. Watson) D. P. Little, Syst. Bot. 31: 473. 2006. Hesperocyparis lusitanica (Mill.) Bartel, comb. nov. Cupressus lusitanica Muill., Gard. Dict., ed. 8. Cupressus no. 3. 1768. 182 Phytologia (April 2009) 91(1) Callitropsis lusitanica (Mill.) D. P. Little, Syst. Bot. 31: 474. 2006. Hesperocyparis macnabiana (A. Murray bis) Bartel, comb. nov. Cupressus macnabiana A. Murray bis, Edinburgh New Philos. J. ser. 2, 1: 293. 1855. Callitropsis macnabiana (A. Murray bis) D. P. Little, Syst. Bot. 31: 474. 2006. Hesperocyparis macrocarpa (Hartw. ex Gordon) Bartel, comb. nov. Cupressus macrocarpa Hartw. ex Gordon, J. Hort. Soc. London 2: 187. 1847. Callitropsis macrocarpa (Hartw.) D. P. Little, Syst. Bot. 31: 474. 2006. Hesperocyparis montana (Wiggins) Bartel, comb. nov. Cupressus montana Wiggins, Contr. Dudley Herb. 1: 161. 1933. Cupressus arizonica Greene var. montana (Wiggins) Little, Madrofio 18: 163. 1966. Cupressus arizonica Greene subsp. montana (Wiggins) A. E. Murray, Kalmia 15: 11. 1985. Callitropsis montana (Wiggins) D. P. Little, Syst. Bot. 31: 474. 2006. Hesperocyparis nevadensis (Abrams) Bartel, comb. nov. Cupressus nevadensis Abrams, Torreya 19: 92. 1919. Cupressus macnabiana A. Murray bis var. nevadensis Abrams (Abrams), Ill. Fl. Pacific States 1: 73. 1923. Cupressus arizonica Greene var. nevadensis (Abrams) Little, Madrofio 18: 164. 1966. Cupressus arizonica Greene subsp. nevadensis (Abrams) A. E. Murray, Kalmia 12: 19. 1982. Callitropsis nevadensis (Abrams) D. P. Little, Syst. Bot. 31: 474. 2006. Hesperocyparis pygmaea (Lemmon) Bartel, comb. nov. Cupressus goveniana Gordon var. “pigmaea” Lemmon, Handb. W. Amer. Cone-bearers. ed. 3, 77. 1895. Cupressus pygmaea (Lemmon) Sarg., Bot. Gaz. (Crawfordsville) 31: 239. 1901. Cupressus goveniana Gordon subsp. pygmaea (Lemmon) A. Camus, [Les Cyprés] Encycl. Econ. Sylvicult. 2: 50. 1914. Callitropsis pigmaea (Lemmon) D. P. Little, Syst. Bot. 31: 474. 2006. Phytologia (April 2009) 91(1) 183 Hesperocyparis sargentii (Jeps.) Bartel, comb. nov. Cupressus sargentii Jeps., Fl. Calif. 1: 61. 1909. Cupressus goveniana Gordon var. sargentii (Jeps.) A. Henry, in Elwes & A. Henry, Trees Great Britain 1173. 1910. Callitropsis sargentii (Jeps.) D. P. Little, Syst. Bot. 31: 474. 2006. Hesperocyparis stephensonii (Jeps.) Bartel, comb. nov. Cupressus stephensonii C. B. Wolf, Aliso 1: 125. 1948. Cupressus arizonica Greene var. stephensonii (C. B. Wolf) Little, Madronio 18: 164. 1966. Cupressus arizonica Greene subsp. stephensonii (C. B. Wolf) A. E. Murray, Kalmia 12: 19. 1982. Callitropsis stephensonii (C. B. Wolf) D. P. Little, Syst. Bot. 31: 474. 2006. ACKNOWLEDGEMENTS This research supported in part with funds from U. S. Fish and Wildlife Service, Grant 814307J011. The findings and conclusions in this article are those of the authors and do not necessarily represent the views of the U.S. Fish and Wildlife Service. Thanks to Tonya Yanke for lab assistance. Thanks to Andrea Schwarzbach and Billie Turner for manuscript reviews. LITERATURE CITED Adams, R. P. 2007. Juniperus maritima, the seaside juniper, a new species from Puget Sound, North America. Phytologia 89: 263- 283. Adams, R. P. 2008. The junipers of the world: The genus Juniperus, 2nd Ed., Trafford Publ., Victoria, BC. Adams, R. P., A. E. Schwarzbach, J. A. Morris and S. Gonzalez Elizondo. 2007. Juniperus compacta, a new species from Mexico (Cupressaceae). Phytologia 89: 361-369. Averyonov, L. V., T. H. Nguyén, D. K. Harder and K. L. Phan. 2002. The history of discovery and natural habitats of Xanthocyparis vietnamensis (Cupressaceae). Turczaninowia 5: 31-39. Brummitt, R. K. 2007. Report on nomenclature committee for vascular plants: Taxon 59: 1289-1296. Camus, A. 1914. Les Cyprés. Encyclopédie économique de Sylviculture 2 Paul Lechevalier, Paris. 184 Phytologia (April 2009) 91(1) Cukovic, D., J. Ehlting, J. A. VanZiffle and C. J. Douglas. 2001. Structure and evolution of 4-coumarate: Coenzyme A Ligase (4CL) gene families. Biol. Chem. 382: 645-654. Debreczy, Z., K. Musial, R. A. Price and I. Racz. 2009. Relationships and nomenclatural status of the Nootka cypress (Callitropsis nootkatensis, Cupressaceae) Phytologia 91(1):140-159 Edwards, S. W. 1983. Cenozoic history of Alaskan and Port Orford Chamaecyparis cedars. Ph.D. Dissertation, University of California, Berkeley. Farjon, A. 1993. Nomenclature of the Mexican cypress or “cedar of Goa,” Cupressus lusitanica Mill. (Cupressaceae). Taxon 42: 81- 84. Farjon, A., 1998. World checklist and bibliography of conifers. Royal Bot. Gard., Kew, London. Farjon, A. 2005. A monograph of Cupressaceae and Sciadopitys. London: Royal Botanic Gardens, Kew. Farjon, A. 2007. In defence of a conifer taxonomy which recognises evolution. Taxon 56: 639-641. Farjon, A., T. H. Nguyén, D. K. Harder, K. L. Phan and L. Averyanov. 2002. A new genus and species in Cupressaceae (Coniferales) from northern Vietnam, Xanthocyparis vietnamensis. Novon 12: 179- 189. Gadek, P. A., D. L. Alpers, M. M. Heslewood and C. J. Quinn. 2000. Relationships within Cupressaceae (sensu lato): a combined morphological and molecular approach. Am. J. Bot. 87: 1044- 1057. Hamberger, B. and K. Hahlbrock. 2004. The 4-coumarate: CoA ligase family in Arabidopsis thaliana comprises one rare, sinapate- activating and three commonly occurring isoenzymes. PNAS 101: 2209-2214. Lazarova, G., Y. Zeng and A. R. Kermode. 2001. Cloning and expression of an ABSCISIC ACID-INSENSITIVE 3(abi3) gene homologue from yellow-cedar (Chamaecyparis nootkatensis). J. Exp. Bot. 53: 1219-1221. Little, E. L., 1970. Names of new world cypresses (Cupressus) Phytologia 20 :429-445. Little, D. P. 2006. Evolution and circumscription of the true cypresses (Cupressaceae: Cupressus) Syst. Bot. 31: 461-480. Phytologia (April 2009) 91(1) 185 Little, D. P., A. E. Schwarzbach, R. P. Adams and C-F. Hsieh. 2004. The circumscription and phylogenetic relationships of Callitropsis and the newly described genus Xanthocyparis (Cupressaceae). Amer. J. Bot. 91: 1872-1881. Mill, R. R. and A. Farjon. 2006. Proposal to conserve the name Xanthocyparis against Callitropsis Oerst. (Cupressaceae). Taxon 50° 229-231. Peng, D. and X-Q. Wang. 2008. Reticulate evolution in Thuja inferred from multiple gene sequences: Implications for the study of biogeographical disjunction between eastern Asia and North America. Molec. Phylogenetics Evol. 47: 1190-1202. Rushforth, K. 2007. Notes on the Cupressaceae in Vietnam. J. Biol. (Vietnam) 29(3): 32-35. Silba, J. 1983. Addendum to revised generic concepts of Cupressus L. (Cupressaceae). Phytologia 52: 349-361. Silba, J. 1994. The trans—Pacific relationship of Cupressus in India and North America. J. Int. Conifer Preserv. Soc. 1(1): 1-28. Silba, J. 1998. A monograph of the genus Cupressus L. J. Int. Conifer Preserv. Soc. 5(2): 1-98. Silba, J. 2005. A monograph of the genus Cupressus L. in the twenty- first century. J. Int. Conifer Preserv. Soc. 12(2): 31-103. Silba, J. 2006. The chronological history and taxonomic variation of the genus Cupressus (Cupressaceae) in India. Acta Bot. Yunnanica 28(5): 469-470. Wolf, C. B. 1948. Taxonomic and distributional studies of the New World cypresses. Aliso 1: 1-250. Xiang, Q. and J. Li. 2005. Derivation of Xanthocyparis and Juniperus from within Cupressus: evidence from sequences of nrDNA internal transcribed spacer region. Harvard Pap. Bot. 9: 375-382 186 Phytologia (April 2009) 91(1) NEW NAMES IN CHAMAESARACHA (SOLANACEAE) James Henrickson Plant Resources Center University of Texas, Austin 78712 ABSTRACT Two new names are proposed for Chamaesaracha: C. arida Henrickson, [C. coronopus auct. non (Moric. ex Dunal) A. Gray] and C. texensis Henrickson [C. conoides auct. non (Moric. ex Dunal) Britt.] Chamaesaracha villosa Rydb. is used for the taxon inclusive of C. crenata Rydb. Phytologia 91(1):186-188 (April, 2009). KEY WORDS: Chamaesaracha arida, C. texensis, Solanaceae. A new name is needed for a distinct taxon that was included within Chamaesaracha coronopus (Moric. ex Dunal) A. Gray by Averett (1973). CHAMAESARACHA ARIDA Henrickson, sp. nov. TYPE: U.S.A. New Mexico: Santa Fe Co., ca. 19 mi. s. of Santa Fe, N.M. on Hwy. 85, 15 Jul 1968, J.-E. Averett & A.S. Tomb 339 (holotype TEX!). Folia caules pedicellique subglabri pilis dispersis basibus latis distaliter furcatis vel ramosis 0.1-0.3 mm longis, segmentis pilorum superiorum brevis latis obtusisque, raro pilis gradatim angustatis simplicibus vel furcatis ad 1.5 mm longis, foliis linearis vel lineari-lanceolatis vel lineari oblanceolatis marginibus undulatis et dentatis vel pinnati-lobatis. Leaves, stems, pedicels subglabrous, with scattered, broad-based, distally forked to branched hairs 0.1-0.3 mm long, the upper hair branches short, blunt, rarely with tapering, simple or distally branched Phytologia (April 2009) 91(1) 187 hairs; leaves linear to linear-lanceolate, linear-oblanceolate, the margins undulate, toothed or pinnately lobed. Study of lectotype and syntype specimens of C. coniodes (Moric. ex Dunal) Britt. at Geneva (G), show that the taxon is characterized by a dense low vestiture of forked-branched hairs. Averett’s (1973) and Rydberg’s (1896) usage of the name for a stipitate glandular species is incorrect and a new name is provided herein for the stiptiate glandular species. CHAMAESARACHA TEXENSIS Henrickson, sp. nov. [Chamaesachara coniodes auct. non (Moric. ex Dunal) Britt.]. Type: U.S.A. Texas: Kinney Co., open rocky soil near the Nueces River, Hy. 334, 17 Apr 1957, D.S. Correll 15965 with R.C. Rollins & K. Chambers (holotype LL!). A Chamaesaracha sordida foliorum juniorum irregulatim dentato- laceratis vel pinnatifidis lobatis integris vel dentatis (non dentatis obtusis vel non profundis paucisque) et tetraploideis (n=24) non diploideis (n=12) dignoscenda. From Chamaesaracha sordida distinguished by the young leaves being irregularly toothed-lacerate to pinnatifid, with entire to toothed lobes (not entire to bluntly or shallowly few toothed) and tetraploid (n=24) (not diploid n=12). I herein combine C. villosa Rydb. with C. crenata Rydb., and use the name C. villosa Rydb. for the inclusive species. Both were published on the same page in Mem. Torr. Bot. Club. 4:368. 1896. ACKNOWLEDGEMENTS Latin was provided by Paul Fryxell. The manuscript was reviewed by T. Wendt and B. Simpson of TEX-LL. 188 Phytologia (April 2009) 91(1) LITERATURE CITED Averett, J. E. 1973. Biosystematic study of Chamaesaracha (Solanaceae). Rhodora 75: 325-365. Rydberg, P. A. 1896. North American species of Physalis and related genera. Mem. Torrey Bot Club 4; 297-374. LOL HA PHYTOLOGIA is published three times a year. Annual domestic individual subscription (3 issues): $30.00. Annual domestic institutional subscription (3 issues): $90.00. Foreign individual subscription $50, Foreign institutional subscriptions $110, send subscription checks to Robert P. Adams, Mgr. Ed., P. O. Box 727, Gruver, TX 79040, USA. Please make checks payable to Phytologia. 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