©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at Septoria epambrosiae sp. nov. on Ambrosia artemisiifolia (common ragweed) David F. Farr & Lisa A. Castlebury Systematic Botany and Mycology Laboratory, USDA, Agricultural Research Service, Beltsville, Maryland 20705-2350 Farr, D. F. & L. A. Castelbury (2001). Septoria epambrosiae sp. nov. on Ambrosia artemisiifolia (common ragweed). - Sydowia 53(1): 81-92. Recently an unidentified Septoria was reported to be pathogenic on Ambrosia artemisiifolia L. (common ragweed) in Hungary and may have potential as a biocontrol agent for this noxious weed. After reviewing the two previously known species of Septoria on Ambrosia (Asteraceae), the Septoria from Hungary is described as a new species, Septoria epambrosiae. A synopsis of the three species of Septoria on Ambrosia is provided. Based on a molecular analyses of the nSSU and complete ITS region of the rDNA, the taxonomic relationships of S. epambrosiae are determined. Septoria epambrosiae clusters with members of the Dothideales with the teleomorph most likely a species of Mycosphaerella. Keywords: Ascomycetes, molecular taxonomy, biocontrol, taxonomy. Ragweed {Ambrosia artemisiifolia L., Asteraceae) is a noxious weed common throughout temperate regions that produces quantities of pollen to which many humans have an allergic reaction, often referred to as hay fever. A species of Septoria Sacc. in Hungary was discovered and tested as a potential biocontrol agent of ragweed (Bohär & Schwarczinger, 1999). Ragweed plants grown in the greenhouse were inoculated with a conidial suspension of this fungus. After 3-4 weeks the infected leaves became completely necrotic and occasionally the entire plants died (Bohär & Schwarczinger, 1999). The possibility of developing this fungus as a biocontrol agent necessitated an accurate identification. The coelomycetous genus Septoria includes about 1,000 described species for which no monographic account exists (Rossman & al., 1987), thus identification is difficult. Hostspecificity in Septoria has been assumed and the few recent studies suggest that this assumption is justified, at least, at the level of host genus (Farr, 1991, 1992; Goodwin & Zismann, 1999). Two species of Septoria have been described from Ambrosia, differing primarily in conidial characteristics; none of these previously known species, however, agree with the potential biocontrol fungus of ragweed. The fungus studied by Bohär & Schwarczinger (1999) and three 81 ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at specimens misidentified as Septoria bacilligera Winter on Ambrosia in BPI are described herein as a new species of Septoria. The taxonomic relationships and the potential teleomorph of this new asexual species were determined by an analysis of sequences of the nSSU and complete ITS region of the rDNA. Materials and methods For microscopic examination material was rehydrated and mounted in 3% KOH. Conidiomata were sectioned at about 10 (.im thick using a freezing microtome. Sections were mounted in lactic acid with cotton blue. Observations of microscopic features were made using a Zeiss Axioplan 2 microscope with both bright-field and fluorescence illumination. Calcofluor was used as the fluorescent dye. Measurements were taken using a digital camera and ImagePro software (Media Cybernetics, Silver Spring, MD). Scatterplots of conidial dimensions were constructed using Systat 7.0 for Windows. The Gaussian bivariate ellipses are graphed for the samples in each plot. The resulting ellipse is centered on the sample means of the x and y variables. The unbiased sample standard deviations of x and y determine its major axes and its orientation is determined by the sample covariance between x and y. A probability of 0.6827 was used to determine the size of the ellipses. Cultures were isolated from nature on Difco Potato Dextrose Agar (PDA). To produce and observe pycnidia in culture the fungus was grown on autoclaved 20-40 x 1-2 mm stems of Medicago (alfalfa) placed on distilled water agar in petri dishes. All isolates were maintained on Difco Corn Meal Agar slants with an alfalfa stick at 4 C and in water cultures (Burdsall & Dorworth, 1994). For the growth studies, cultures were grown on Difco Czapek Solution Agar (CZ), Difco Corn Meal Agar (CM) and PDA at 25 C in the dark. Color names and numbers were determined using Kornerup & Wanscher (1978). DNA was extracted from lyophilized mycelium using the Qiagen Plant DNeasy kit (Qiagen Inc., Chatsworth, CA). The internal transcribed spacer (ITS) regions 1 and 2, including the 5.8S rDNA, were amplified in 50 f.iL reactions on a GeneAmp 9700 thermal cycler (Perkin Elmer/ABI, Foster City, CA) under the standard reaction conditions with 15 ng of genomic DNA, 2.5 units Amplitaq Gold IM (PE Biosystems, Foster City, CA), 25 pmoles each of primers ITS5 and ITS4 (White & al., 1990) and the supplied PCR buffer. The thermal cycler program was as follows: 10 min at 95 C followed by 35 cycles of 30 s at 94 C, 30 s at 55 C, 1 min at 72 C, with a final extension period of 10 min at 72 C. The nuclear small subunit ribosomal (nSSU) RNA genes were amplified under the same 82 ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at conditions, using primers NS1 and NS8 (White & al., 1990). Following amplification, the PCR products were purified with QIAquick (Qiagen Inc., Chatsworth, CA) columns according to the manufacturer's instructions. Purified PCR products were sequenced on an ABI 310 automated DNA sequencer using the BigDye Terminator kit (PE Biosystems) according to the manufacturer's instructions. Sequencing primers were as follows: ITS5 and ITS4 for the ITS region and NS1, NS3, NS4, NS5, NS6, and NS8 for the nSSU genes. The ITS sequence has been deposited at GenBank with the number AF 279582 and the SSU sequence with the number AF 279583. The resulting sequences were edited, joined, and aligned using the Sequencher software program (Gene Codes Corp, Ann Arbor, MI). Additional sequences of Septoria, Mycosphaerella, Capnodium Mont., Communitispora Ramaley and Dothidea Fr. were obtained from GenBank. The sources of these sequences along with the original isolate number followed by the GenBank accession number are: Crous & al. (1999), M. africana Crous & M. J. Wingf. (STE-U 794, AF173314), M. ellipsoidea Crous & M. J. Wingf. (STE-U 1225, AF173303), M. ellipsoidea (STE-U 1224, AF173302), M. fragariae (Tul.) Lindau (STE-U 656, AF173312), M. juvenis Crous & M. J. Wingf. (STE-U 1005, AF173299), M. keniensis Crous & T. Coutinho (STE-U 1084, AF173300), M. marksii Carnegie & Keane (STE-U 935, AF173316), M. molleriana (Thüm.) Lindau (STE-U 784, AF173313), M. molleriana (STE-U 1214, AF173301), M. parkii Crous & M. J. Wingf. (STE-U 353, AF173311), M. suttoniae Crous & al. (STE-U 1346, AF173306), M. tasmaniensis Crous & M. J. Wingf. (STE-U 1457, AF173307); Goodwin & Zismann (Unpublished), M. citri Whiteside (Fellsmere, AF181703), M. citri (GS8, AF181704), M. fijiensis M. Morelet (ATCC 22116, AF181705), M. graminicola (Fuckel) Schroet. (IPO323, AF181692), M. graminicola (Tl, AF181693), M. graminicola (T48, AF181694), M. musicolaJ. L. Mulder (ATCC 22115, AF181706), S. passerinii Sacc. (P70, AF181700), S. passerinii (P78, AF181699), S. passerinii (P83, AF181698), S. passerinii (ATCC 26516, AF181697), S. passerinii (ATCC 26515, AF181696), S. passerinii (ATCC 22585, AF181695), S. passerinii (P71, AF181701); Hoog & al. (Unpublished), Capnodium coffeae Pat. (CBS 147.52, AJ244239) C. salicinum Mont. (CBS 131.34, AJ244240), Comminuntispora agavaciensis Ramaley (AJ244250, CBS 619.95); Jacobs & Rehner (1998), D. hippophaeos (Pass.) Fuckel (CBS 186.58, AF027763), D. insculpta Wallr. (CBS 189.58, AF027764); Schenck & Frey (not published), M. dearnessii Barr (MDUS1, AF211196), M. dearnessii (MD038, AF211195), M. dearnessii (MD002, AF211194), M. pini Rostr. (MP002, AF211197), M. tassiana (De Not.) Johanson (CBS 111.82, AF 238469); Ueng & Subramanian (Unpublished), M. graminicola (Unknown, U77363). 83 ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at Alignments were manually adjusted using MacClade 3.0 (Maddison & Maddison, 1992) and GeneDoc 2.5 (Nicholas & Nicholas, 1997). Maximum parsimony trees were inferred using the heuristic search (simple addition sequence) and branch swapping (tree bisection-reconnection) options of PAUP 4.0b4a (Swofford, 1999). All molecular characters were unordered and given equal weight during analysis. Relative support for the branches was estimated with 100 bootstrap replications (Felsenstein, 1985). Description Septoria epambrosiae D. F. Farr, sp. nov. - Figs. 1-7. Pycnidia globosa, fusca, 48-104 |.im diametro, ostiolata; cellulis conidiogenis pyriformibus, hyalinis, 3-5x7-17 \xm, supera elongatio cellulae parietibus irregulariter incrassatis; conidiogenesis holoblastica, sympodialis; conidia hyalina, recta vel parum arcuata, interdum flexuosa, 17-41 x 1.0-2.3 |.im, 0-4 septata, basi truncata, gradatim dimidiata versus rotundatum apicem. H o l o t y p e . - HUNGARY. Pest County, parasitic on leaves of Ambrosia artemisiifolia, 1997, Bohär G. & I. Schwarczinger, BPI 747030, ex-type culture ATCC MYA-970. L e s i o n s 0.5-5 mm in longest dimension, angular to irregular, white with a brown margin or more uniformly brown without a distinct margin, with 1 to several, scattered, immersed pycnidia. P y c n i d i a 48-104 (am diameter, globose, brown-black, ostiolate, wall of one to two layers of pale brown cells, outer layer darker than inner layer, cell walls of outer layer slightly thickened (Fig. 2). C o n i d i o g e n e s i s holoblastic, sympodial. - C o n i d i o g e n o u s cells lining inner wall, 3-5 x 7-17 |.im, pyriform, hyaline, upper portion covered with irregular wall thickenings, number of scars 2-6 per cell (Figs. 3, 4). - C o n i d i a 17-41 urn (x = 27.4, SD = 2.6, number = 130) x 1.0-2.3 (x = 1.7, SD = 0.15), 0-4 (x = 2.1, SD = 0.83) septate, straight to slighty falcate, occasionally flexuous, base truncate, gradually tapering to a rounded apex, hyaline (Figs. 6, 7). P y c n i d i a of ex-type culture forming on alfalfa have one or more beaks up to 110 f.im long x 10 j.im wide, occasionally branching (Fig. 5). - P y c n i d i a l w a l l s thicker than on leaves, with 3-6 layers of dark brown cells; cell walls somewhat thickened; innermost 1 or 2 layers hyaline (Fig. 1). - Conidia longer, more flexuous than on leaves, 21-46 (im (x = 32.0, SD = 8.7, number = 49) x 1.1-1.8 urn (x 1.4, SD = 0.16), 0-5 septate (x = 2.4, SD = 0.81). C u l t u r a l c h a r a c t e r i s t i c s of ex-type culture, growth after 6 weeks: PDA: - Colony 44 mm diam, brown (6E5) to black with white cottony mycelium around plug, margin uneven to feathery, 84 ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at 1 Figs. 1-4. Septoria epambrosiae. - 1. Longitudinal section of conidiomata on alfalfa stem in cultures. - 2. Longitudinal section of conidiomata on Ambrosia leaves. - 3-4. Conidiogenous cells viewed with fluorescence illumination. - Scale bars: 1-2 = 20 (.im; 2-3 = 10 (im. 85 ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at Figs. 5-9. - 5-7. Septoria epambrosiae. - 5. Conidiomata on alfalfa twigs in culture. - 6. Conidia viewed with bright-field illumination. - 7. Conidia viewed with fluorescence illumination. 8-9. - Septoria ambrosiicola. - 8. Conidiogenous cells viewed with fluorescence illumination. - 9. Conidia viewed with bright-field illumination. - Scale bars: 5 = 20 (.im; 6-9 = 10 urn. pycnidia common. CM: - Colony 30 mm diam, pale brown (5D5) with no surface mycelium, margin even, sparse pycnidia around plug. CZ: Colony 33 mm diam, dark brown (6F6) to black, dark brown felty mycelium around plug, margin uneven, scattered pycnidia. H a b i t a t . - Parasitic on leaves of Ambrosia (Asteraceae). Known hosts. - Ambrosia trifida L. and A. artemisiifolia L. Known d i s t r i b u t i o n . - Europe (Hungary) and North America (Canada, United States-Wisconsin). A d d i t i o n a l s p e c i m e n s e x a m i n e d . - CANADA: Ontario, London, 1910 Sep 19, on A. trifida leaves, J. Dearness, as Septoria bacilligera, BPI 377153; Ontario, London, 1923 Sep, on A. trifida, J. Dearness, as Septoria bacilligera, BPI 377156. - UNITED STATES: Wisconsin, Jefferson Co., Milford, 1948 May 15, on A. trifida leaves, H. C. Greene H. C, as Septoria bacilligera, BPI 377155. 86 ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at Discussion The pycnidial genus Septoria, based on the type species S. cytisi Desm., has conidiogenous cells that are holoblastic, determinate or indeterminate, with a limited number of sympodial proliferations and each locus with a broad, flat, unthickened scar (Sutton, 1980). The conidia are characterized as hyaline, multiseptate and filiform. Septoria epambrosiae agrees with this generic description. Two species of Septoria were previously described on Ambrosia; the fungus on Ambrosia from Hungary, however, is determined to be different from these species. The principal characteristics of the three species of Septoria on Ambrosia are summarized in Tab. 1. Only the original description of Septoria ambrosiae Hemmi & Naito in Naito (1940) was available while two specimens of S. ambrosiicola Speg. (1910) were examined {S. ambrosiicola - UNITED STATES: Kansas, Rooks Co., 1895 Sep 27, on A. trifida, Elam Bartholomew, misidentified as Septoria bacilligera, BPI 377161; Texas, Austin, 1916 Jun 17, B. C. Tharp, on A. aptera, BPI 376564). Conidia of Septoria ambrosiae have a greater number of septa than those of S. epambrosiae and S. ambrosiicola Speg. While Spegazzini (1910), in the original description, and later Tharp (1917) described the conidia of S. ambrosiicola as being non-septate, conidia from the Tharp specimen at BPI were 0-5 septate and agreed in all other aspects with the description of S. ambrosiicola (Figs. 8, 9). Conidia of S. ambrosiae are wider than those of S1. ambrosiicola and S. epambrosiae. Conidia of S. ambrosiicola are longer and somewhat wider than those of S. epambrosiae as shown by scatterplots of conidial length and width measurements (Fig. 10). In addition, conidia of S. epambrosiae are straight to slightly falcate while those of S. ambrosiae and S. ambrosiicola are flexuous or undulate. A striking feature of S. epambrosiae and S. ambrosiicola is the numerous knobby protuberances on the conidiogenous cells; these represent the scars left when the conidia are discharged (Figs. 3, 4, 8). Sympodial conidiogenesis has been reported from other Septoria sp. (Sutton & Pascoe, 1987; Verkley & Priest, 2000) but is uncommon in the genus (Sutton, 1980; Farr, 1991; Farr, 1992). The number of scars per conidiogenous cell varied from 2-6 or more in specimens of both S. epambrosiae and S1. ambrosiicola suggesting that this is not a character useful for defining species in Septoria. A GenBank BLAST search (Altschul & al., 1990) of the small SSU rDNA of S1. epambrosiae revealed similarities to Coccodinium bartschii, Dothidea insculpta and D. hippophaeos. A cladogram based on the nSSU rDNA shows S. epambrosiae grouping with the Dothideales in a clade with the taxa mentioned above (DePriest, 87 ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at 00 oo Tab. 1. - Comparison of species of Septoria on Ambrosia. Name Host Lesion I'vcnidia Conidia Septoria epambrosiae Farr Ambrosia trifida and A. artemisiifolia Angular to irregular, white with brown margin or entirely brown Brown-black, globose, 48-104 urn diam 17-41 (x = 27.4) x 1.0-2.3 (x = 1.7) urn, 0-4- (x = 2.1) septate, straight to slightly falcate Septoria ambrosiae Hemmi & Naito Ambrosia artemisiifolia Oblong to irregular, sometimes confluent, at length occupying a large part of the leaf, black brown Brown, globose, 95.65- 40-64 x 2-2.6 urn, 3-7-septate, attenuate-acutate, filiform, straight 121.70 x 86.95-121.70 (.im diam. membranous, to flexuous epiphyllus Septoria ambrosiicola Speg. Ambrosia aptera, A. trifida, originally described on A. scabrac White above, tan below, tanbrown margin. 0.5-2 mm. angular to suborbieular Brown-black, globose, 57-123 (.im diam 21-62 (x = 44) x 1.7-2.6 (x = 2) pm, 0-5-(x = 2.3) septate, subarcuate to flexuous ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at 70 60 E 50 —i c 40 <D 30 20 - 10 1.5 2.0 Width in [jm 2.5 3.0 Fig. 10. - Scatterplot of the conidial length and width of Septoria epambrosiae (x = BPI 377153, A = BPI 377156, + = BPI 377155, < = BPI 747030 and Septoria ambrosiicola (V = BPI 377161, O = BPI 376564). pers. comm.). A GenBank BLAST search of the ITS sequence placed S. epambrosiae, among species of Mycosphaerella (Dothideales). A cladogram of ITS sequences of species of Mycosphaerella and related species in the Dothideales was constructed to investigate the relationships of S. epambrosiae (in Fig. 11). If known, the anamorph for the species of Mycosphaerella is listed in the cladogram. Based primarily on sequences obtained and analyzed by Crous & al. (1999, 2000), S. epambrosiae clusters with species of Mycosphaerella having Pseudocercospora Speg. and Mycovellosiella Rangel anamorphs. Thus, the teleomorph of S1. epambrosiae, if found, would most likely belong to Mycosphaerella. A number of Septoria species have been reported to have Mycosphaerella teleomorphs (Rossman & al., 1987; Farr & al., 1989; Brzustowski, 2000). Septoria tritici Rob., the anamorph of M. graminicola, and S. passerini, cluster within the main Mycosphaerella clade but are not particularly close to S. epambrosiae. Within Mycosphaerella, the known anamorphs are classified in both hyphomycetous genera such as Cercospora Fresen. and ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at S. passerinii AF181701 S. passerinii AF181700 S. passerinii AF181699 S. passerinii AF181698 S. passerinii AF181697 S. passerinii AF181695 L S. passerinii AF181696 100 M. graminicola AF181694 M. graminicola AF181693— Soptoria 55 M. graminicola AF181692 M. graminicola U77363 • M. fragariae AF173312— Ramularia • M. citri AF181704- Stenella I M. citri AF181703 87|M. africana AF173314 96fl M- keniensis AF173300 M. pini AF211197- Dothistroma M. dearnessii AF211196 100 97 M. dearnessii AF211195— Lecanosticta M. dearnessii AF211194 55 100 i - M. ellipsoidea AF173303- Uwebraunia L- M. ellipsoidea AF173302 100 i — — M. musicola AF181706— Pseudocercospora •M. fijiensis AF81705 83 57 100 S. epambrosia AF279582 —— M. tasmaniensis AF173307— Mycovellosiella • M. marksii AF173316- Stenella •M. parkii AF173311- Stenella M. juvenis AF173299— Uwebraunia M. molleriana AF173313- Colletogloeum 100 62 70 M. molleriana AF173301 100 56 M. suttonlae AF173306— Phaeophleospora Ca. coffeae AJ244239 • Ca. salicinum AJ244240-Fumagospora • Co. agavaciensis AJ244250 • M. tassiana MTA238469- Cladosporium - D. hippophaeos AF027763 - D. insculpta AF02*764~ Scirrhla ^^— 10 changes Fig. 11. - One of six most parsimonious trees from analysis of the complete ITS region of rDNA. Bootstrap values greater than 50% are listed above the branches. Length = 860 steps, CI = 0.492, RI = 0.714, RCI = 0.351, alignment = 568 total characters, 93 ambiguously aligned positions excluded. 199 parsimony informative characters analyzed. 90 ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at Passalora Fr. and coelomycetous genera, such as Colletogloeum Petr. and Phaeophleospora Rangel. References Altschul, S. E, W. Gish, W. Miller, E. W., Myers & D. J. Lipman (1990). Basic local alignment search tool. - J. Mol. Biol. 215: 403-410. Bohär, Gy., & I. Schwarczinger (1999). First report of a Septoria sp. on common ragweed (Ambrosia artemisiifolia) in Europe. - PL Dis. 83: 696. Burdsall Jr., H. H. &, E. B. Dorworth EB. 199(1994). Preserving cultures of wooddecaying Basidiomycotina using sterile distilled water in cryovials. Mycologia 86: 275-280. Brzustowski, J. (2000). Query the Anamorph/Holomorph Connections Database. HYPERLINK „http://www.biology.ualberta.ca/jbrzusto/anateleo/anatel.html" (May 2000). Crous, P. W., A. Aptroot, J.-C. Kang, U. Braun & M. J. Wingfield (2000). The genus Mycosphaerella and its anamorphs. - Stud. Mycol. 45: 107-121. , L. Hong, M. J. Wingfield & B. D. Wingfield. (1999). Uwebraunia and Dissoconium, two similar anamorph genera with distinct teleomorph affinity. - Sydowia 51: 155-166. Farr, D. F. (1991). Septoria species on Cornus. - Mycologia 83: 611-623. , (1992). Species of Septoria on the Fabaceae, subfamily Faboideae, tribe Genisteae. - Sydowia 44: 13-31. , G. F. Bills, G. P. Chamuris & A. Y. Rossman (1989). Fungi on Plants and Plant Products in the United States. - American Phytopathological Society, St. Paul, Minnesota. 1252 pp. Felsenstein J. (1985). Confidence limits on phylogenies: an approach using the bootstrap. - Evolution 6: 227-242. Goodwin, S. B. & V. L. Zismann. (1999). Septoria passerinii is closely related to the wheat pathogen Mycosphaerella graminicola. - In: van Ginkel, M., A. McNab & J. Krupinsky, (eds.). Septoria and Stagonospora diseases of cereals: a compilation of global research. CIMMYT, Mexico, D.E: 34-36. Jacobs, K. A. & S. A. Rehner (1998). Comparison of cultural and morphological and ITS sequences in Botryosphaeria and related taxa. - Mycologia 90: 601-610. Kornerup A. & J. H. Wanscher (1978). Methuen handbook of colour. - Methuen & Co., London. 243 pp. Maddison W. P. & D. R. Maddison (1992). MacClade, version 3.0. - Sinauer Associates, Inc., Sunderland, Massachusetts. Naito, N. (1940). Studies on Septorioses of plants. VII New or noteworthy species of Septoria found in Japan. - Mem. Coll. Agr, Kyoto Imp. Univ. 47: 31-43. Nicholas, K. B. & H. B. Nicholas, Jr. (1997). GeneDoc: a tool for editing and annotating multiple sequence alignments. - Distributed by the authors. Rossman, A. Y, M. E. Palm, & L. J. Spielman ((1987)). A Literature Guide for the Identification of Plant Pathogenic Fungi. - American Phytopathological Society, St. Paul, MN. 252 pp. Spegazzini, C. (1910). Mycetes argentinenses (series V). - An. Mus. Nac. Buenos Aires 20: 329-467. Sutton, B. C. (1980). The Coelomycetes. - Commonwealth Mycological Institute, Kew, England. 696 p. & I. G. Pascoe (1987). Septoria species on Acacia. - Trans. Brit. Mycol. Soc. 89: 521-532. 91 ©Verlag Ferdinand Berger & Söhne Ges.m.b.H., Horn, Austria, download unter www.biologiezentrum.at Swofford D. (1999). PAUP*, version 4.04b. - Sinauer Associates, Sunderlund, Massachusetts. Tharp, B. C. (1917). Texas parasitic fungi. - Mycologia 9: 105-124. Verkley, G. J. M. & M. J. Priest (2000). Septoria and similar coelomycetous anamorphs of Mycosphaerella. - Stud. Mycol. 45: 123-128. White, T. J., T. Bruns, S. Lee & J. W. Taylor (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. - In: M. A. Innis, D. H. Gelfand, J. J. Sninsky & T. J. White (eds.) PCR protocols - a guide to methods and applications. - Academic Press, San Diego, California: 315^322. (Manuscript accepted 23"' September 2000) 92 ZOBODAT - www.zobodat.at Zoologisch-Botanische Datenbank/Zoological-Botanical Database Digitale Literatur/Digital Literature Zeitschrift/Journal: Sydowia Jahr/Year: 2001 Band/Volume: 53 Autor(en)/Author(s): Farr David F., Castlebury Lisa A. Artikel/Article: Septoria epambrosiae sp. nov. on Ambrosia artemisiifolia (common ragweed). 81-92