Relationships among the perennial, endemic umbellifers of western NA

1. Relationships among the perennial, endemic umbellifers of western NA Stephen R. Downie Feng-Jie Sun Ronald L. Hartman

2. Meet Ph.D. Student Feng-Jie Sun!A phylogenetic study of Cymopterus and related genera

3. Meet NA Umbellifer Cymopterus (the spring-parsleys) • Perennial, taprooted herbs • Low-growing, acaulescent • Ternately-cleft, compound or dissected leaves • Spreading compound umbel • Yellow or white flowers, opening very early in spring Cymopterus purpureus • Mericarps conspicuously winged • (kyma=wave; pteron=wing) Cymopterus montanus (RM Herbarium Specimen)

4. BONAP (TAMU) Number of species per state: 1-4 5-8 9-12 13-15 16-18 Cymopterus (35 spp.) is distributed primarily throughout the Intermountain and Montane regions of western NA

5. Probable relationship of Cymopterus and its allies A monograph of Cymopterus including a critical study of related genera (Mathias 1930) UC TV

6. Aletes (6) Glehnia (1) Harbouria (1) Lomatium (78) Musineon (4) Neoparrya (1) Oreonana (3) Oreoxis (4) Orogenia (2) Podistera (4) Pseudocymopterus (2) Pteryxia (4) Shoshonea (1) Tauschia (10) Cymopterus is putatively allied to other indigenous, perennial umbellifers of western NA (number of species according to Kartesz, 1994)

7. Dorsal ribs of the fruit (or at least one of them) generally more or less conspicuously winged, rarely all wingless …………... Cymopterus Dorsal ribs of the fruit wingless or nearly so ………….. Lomatium Cronquist (1997), treatment of Apiaceae for Intermountain Flora

8. lateral wings dorsal ribs Cymopterus lapidosus Several species of Cymopterus lack dorsal wings and their fruits show similarities to those of typical Lomatium C. corrugatus C. deserticola C. douglassii C. lapidosus C. megacephalus C. newberryi C. ripleyi C. williamsii Cymopterus williamsii Hartman & Constance (1985) “the distinction between Cymopterus and Lomatium is subject to failure” “Both genera are of considerable size, and our taxonomic understanding would not be improved by combining them into one.” Cronquist, 1997, Intermountain Flora

9. Pseudocymopterus, “one of the most complex situations in the family” (Mathias 1930) Short, stiff pubescence at top of peduncle Dorsal wings range from well-developed to low ridges, or are completely absent Plants show similarities to Cymopterus & Lomatium Cronquist, 1997

10. Figs. 1-14 Fig. 15 Fig. 16 Fig. 17 Fig. 18 Fig. 19 Fig. 20 Fig. 21 Figs. 22-23 C. fendleri C. cinerarius C. globosus C. megacephalus C. deserticola C. coulteri C. corrugatus C. acaulis C. newberryi Median cross-sections of mature mericarps of Cymopterus (Mathias, 1930)

11. Median cross-sections of mature mericarps (Mathias, 1930) Pseudocymopterus P. montanus P. davidsonii P. anisatus P. hendersonii P. humboldtensis Pteryxia P. terebinthina var. californica var. foeniculacea var. albiflora var. calcarea P. petraea Aulospermum A. panamintense A. duchesnense A. minimus A. rosei A. aboriginum Aletes humilis

12. Taxonomic Confusion Surrounds Cymopterus! Coulter & Rose (1900) and Mathias (1930) recognized many small, generically distinct elements (e.g., Aulospermum, Phellopterus, Pseudocymopterus, Rhysopterus) UCTV Jones (1908) recognized multiple sections and subgroups within a highly variable and expanded genus Cymopterus Mathias & Constance (1944-1945) submerged Aulospermum, Phellopterus and most Pseudocymopterus into Cymopterus (32 spp.), while maintaining Rhysopterus as distinct The Regents of the University of California

13. Contemporary Treatments Recognize 35-45 Species in Cymopterus Cronquist (1961, 1997) included Oreoxis, Pseudocymopterus and Pteryxia within a broadly circumscribed Cymopterus (with ca. 45 species) “The taxonomic definition of Cymopterus is a vexed question.” Hunt Institute Kartesz (1994), whose checklist of NA Apiaceae was influenced by Lincoln Constance, maintained these three genera as distinct, and recognized 35 species in Cymopterus The Regents of the University of California

14. Relationships of Lomatium among other genera of Apiaceae (Plant Syst. Evol., Gilmartin & Simmons, 1987) Phylogram derived from MP analysis of 10 morphological characters (data set reanalyzed to include Lomatium alliance and 3 outgroup taxa) • Attempted to discern sister taxon to Lomatium • 88 NA genera clustered into 7 phenetic alliances based on development of a stylopodium, presence of a carpophore, and direction of fruit flattening • Monophyly of all genera assumed

15. Objectives • To evaluate the monophyly of Cymopterus • To determine the phylogenetic relationships of the elements that currently comprise Cymopterus with other perennial, endemic apioid umbellifers of western NA • To evaluate the monophyly of these perennial, endemic umbellifers and to ascertain its sister group

16. Relationships among NA Apioideae MP Strict Consensus Tree Showing Tribes and Major Clades • 78 genera (333 spp.) of subfamily Apioideae native or naturalized in NA (north of Mexico; Kartesz, 1994) • All but 3 genera included in molecular analyses to date • Approximately 30 NA apioid genera in Angelica and Arracacia clades

17. Materials and Methods 159 Accessions Examined, including all species and infraspecific taxa of Aletes, Cymopterus, Glehnia, Musineon, Oreonana, Oreoxis, Orogenia, Podistera, Polytaenia, Pseudocymopterus, and Pteryxia. Also, Lomatium (30 accessions and Tauschia (5 accessions), and the monotypic Harbouria, Neoparrya, and Shoshonea. As putative outgroups, we examined Angelica, 3 representatives of the meso-American Arracacia clade, and several members of the perennial, circumboreal genera Seseli, Selinum, and Peucedanum. The trees were rooted with Aethusa. • Phylogenetic Analyses: • Maximum Likelihood (PAUP*) • Maximum Parsimony (PAUP*) • Bayesian Inference (MrBayes)

18. Combined (ITS-1 & ITS-2) 427-441 458 184 168 (37%) 106 37 16 12.1% Sequence Characteristic Length variation (bp) No. of aligned positions No. of positions constant No. of positions informative No. of positions autapomorphic No. of alignment gaps No. of gaps parsimony informative Maximum sequence divergence ITS-1 206-218 225 91 89 45 21 11 12.5% ITS-2 220-226 233 93 79 61 16 5 13.1% Multiple alignment of 159 ITS-1 and ITS-2 DNA sequences

19. Basal Relationships: Monophyly of Perennial, Endemic NA Apioideae MP Strict Consensus Maximum Likelihood

20. Glehnia, “a single species complex occurring on both shores of the northern Pacific” Peucedanum japonicum Angelica hendersonii Charles Webber CA Academy Sciences Br. Alfred Brousseau Saint Mary’s College Glehnia littoralis subsp. leiocarpa subsp. littoralis subsp. leiocarpa Br. Alfred Brousseau St. Mary’s College Mathias, 1928 Hiroe, 1962)

21. M. Schlessman & F. Barrie • Floral protogyny • Nectary morphology • Biogeography • Habitat • Flowering time • Variation in patterns of sex expression ) Evidence for monophyly Overlapping patterns of similar fruit morphology Low growing herbaceous perennials Perennials Obsolete stylopodium (except in Podistera) Largely western NA endemics (north of Mexico) Podistera C. multinervatus (Cronquist 1997) Montane or alpine Xerophytic or semixerophytic habit (occur in dry, sandy or alkaline habitats) ) Pteryxia terebinthina (CalPhotos) Cymopterus globosus (CalPhotos)

22. Cymopterus, Lomatium, and Pteryxia are each highly polyphyletic ML analysis of 135 accessions (18 genera) of NA Apioideae Cymopterus n=41 Lomatium n=30 Pteryxia n=10

23. Maximum Parsimony, Maximum Likelihood, & Bayesian Trees all reveal similar major clades, but resolution within and among them is poor 37 taxa in Clade: Cymopterus (13) Lomatium (16) Oreonana (3) Pteryxia (3) Orogenia (2) Cymopterus and Lomatium are highly polyphyletic, with their species inextricably linked with those of Aletes, Musineon, Oreonana, Oreoxis, Orogenia, Podistera, Pseudocymopterus, Pteryxia, and Tauschia ML analysis 135 accessions (18 genera)

24. Many western NA apioid genera are not monophyletic Additional steps required to force monophyly No. constrained accessions Genus Aletes Cymopterus Lomatium Musineon Oreonana Oreoxis Orogenia Podistera Pseudocymopterus Pteryxia Tauschia 9 41 30 4 3 4 2 5 2 10 5 17 35 45 7 3 15 1 7 4 14 19 (unconstrained MP analysis resulted in minimal length trees of 881 steps)

25. ML analysis 135 accessions (18 genera) Thaspium, Zizia and Polytaenia are each monophyletic and unite as a clade Zizia aurea Zizia Polytaenia BONAP (TAMU) Distribution Maps Thaspium

26. ML analysis 135 accessions (18 genera) Cymopterus sect. Phellopterus Nutt. ex Torr. & A. Gray The only species group of Cymopterus that is supported, in part, by molecular evidence Cymopterus bulbosus Large, showy bractlets that are basally connate and prominently veined Cymopterus constancei Showy bractlets Ronald Hartman Brittonia 52: 137. 2000

27. Do additional data from the chloroplast genome provide increased resolution of relationships? 27 species (12 genera) examined, including: Cymopterus (6 spp.) Lomatium (8 spp.) Angelica to root the tree • Loci Examined: • ITS • rps16 intron • trnF-trnL-trnT Maximum Parsimony (PAUP*)

28. Multiple alignment of 27 DNA sequences from three loci (separately & combined) ITS 437-439 444 7 321 42 74 6 3 6.7% >20,000 196 0.4955 0.5912 rps16 911-976 997 20 902 34 41 13 4 3.2% 6 94 0.6604 0.8583 trnF-L-T 1693-1816 1884 46 1721 42 75 27 7 1.7% 220 145 0.6769 0.8727 Combined 3096-3229 3325 73 2944 118 190 46 14 2.6% 16 478 0.4926 0.6783 Length variation No. of aligned positions No. of positions eliminated No. of positions constant No. of positions informative No. of positions autapomorphic No. of alignment gaps No. of gaps parsimony informative Maximum sequence divergence No. of minimal length trees Length of shortest trees Consistency Index Retention Index

29. Strict consensus trees resulting from MP analysis of ITS and rps16 intron sequence data ITS rps16 intron

30. Combined Molecular trnF/L/T Strict consensus trees resulting from MP analysis of trnF/L/T and combined molecular data

31. Summary • The perennial, endemic apioid umbellifers of NA (north of Mexico) constitute a weakly supported monophyletic group • Sister to this clade are several groups of taxa, including the genus Angelica and the meso-American Arracacia clade • Many western NA genera are not monophyletic, and the two largest genera (Cymopterus & Lomatium) are grossly polyphyletic • Elements of Cymopterus are inextricably linked with Aletes, Lomatium, Musineon, Oreonana, Oreoxis, Orogenia, Podistera, Pseudocymopterus, Pteryxia, and Tauschia

32. Future Plans • Greatest resolution of relationships was obtained by including all molecular data in a simultaneous analysis. Additional molecular data will be procured • Cladistic analysis of morphological data will be done in an effort to uncover synapomorphies useful for generic determination. The traditional emphasis placed on fruit characters has led to highly artificial assemblages of species • If future studies support the conclusions presented herein, and if further resolution of relationships can be achieved, radical changes to the prevailing classification will be required

33. Acknowledgements • NSF Grants 9407712 & 0089452 • John R. Laughnan Travel Award • Herbert H. Ross Memorial Fund • Francis M. and Harlie M. Clark Research Support Grants • Program in Ecology & Evolutionary Biology Travel Grants and Summer Research Fellowship • UIUC Graduate College Dissertation Travel Grant • UIUC Graduate College Dissertation Completion Fellowship • Herbarium curators for access to specimens Deborah S. Katz-Downie

34. Ron Hartman Feng-Jie Sun