Molecular Phylogenetics and Taxonomy of the Genus Thysanocarpus (Brassicaceae)

Patrick J. Alexander , 1 , 4 Michael D. Windham , 2 Rajanikanth Govindarajulu , 1 Ihsan A. Al-Shehbaz , 3 and C. Donovan Bailey 1 1 BiologyProof Department, MSC 3AF, New Mexico State University,Only Las Cruces, New Mexico 88003, U. S. A. 2 Department of Biology, 139 Biological Sciences (Box 90338). Duke University, Durham, North Carolina 27708, U. S. A. 3 Department of Asian Botany, Missouri Botanical Garden, P.O. Box 299, St. Louis, Missouri 63166, U. S. A. 4 Author for correspondence ( [email protected] ); first and second authors contributed equally.

Communicating Editor: Daniel Potter

Abstract—The relationships and taxonomy of the genus Thysanocarpus (Brassicaceae) are reassessed based on molecular phylogenetic analyses of nuclear ribosomal (ITS) and chloroplast (trnL-F ) sequences and a critical re-examination of morphology and nomenclatural types. Based on these results, Thysanocarpus is well-supported as a member of tribe Thelypodieae, but no illuminating phylogenetic structure is found within the tribe. The independent origin of similar fruit morphology in Thysanocarpus and Athysanus is confirmed. Within Thysanocarpus , seven species are recognized: T. conchuliferus, T. curvipes, T. desertorum, T. erectus, T. laciniatus, T. radians, and T. rigidus comb. nov. Thysanocarpus laciniatus is found to have originated through hybridization. However, T. desertorum and T. rigidus, which have previously been included within T. laciniatus (as T. laciniatus var. hitchcockii and T. laciniatus var. rigidus, respectively), do not share that species’ hybrid origin and are distinct both phylogenetically and morphologically. Within T. curvipes, five subspecies are recognized: T. curvipes subsp. amplectens comb. nov., T. curvipes subsp. curvipes , T. curvipes subsp. elegans comb. nov., T. curvipes subsp. longistylus comb. nov., and T. curvipes subsp. eradiatus comb. Nov. Thysanocarpus curvipes subsp. elegans and T. curvipes subsp. longistylus form clades in ITS and/or trnL-F cladograms as well as showing morphological distinction. The remaining three subspecies are recognized based on a combination of morphology and geography. Keywords— Fruit morphology , hybrid speciation , nuclear ribosomal ITS , Thelypodieae, trnL-F .

Thysanocarpus Hook. is a genus of annuals in the fam- to Thysanocarpus (Bailey et al. 2006; Mummenhoff et al. 2005; ily Brassicaceae, found from extreme southwestern British Warwick et al. 2008 ). Columbia south through the western United States to north- The goals of the present study are threefold. First, a family- western Mexico. The genus is most diverse in California, wide phylogenetic analysis is presented to test the mono- where all but one of the species (T. erectus S. Wats., a Baja phyly of Thysanocarpus and establish its position within California endemic) are found. In the most recent synoptic Brassicaceae, including further confirmation of the separation treatment of the genus, Rollins (1993) recognized five species of Thysanocarpus and Athysanus . Second, infrageneric phylo- with no named infraspecific taxa. However, variation within genetic analyses within Thysanocarpus are used to refine our several of these taxa is substantial, and in the earlier treat- understanding of the number of taxa in the genus and their ment of California taxa, Munz (1959) recognized four variet- relationship with each other. Third, a revised taxonomy of the ies within T. curvipes Hook. and five within T. laciniatus Nutt. genus is presented based on both the results of infrageneric ex Torr. & A. Gray. Thysanocarpus is currently placed in the phylogenetic analyses and a re-examination of morphology. tribe Thelypodieae (formerly Schizopetaleae; see Beilstein et al. 2008 and Warwick et al. 2009 ) based on morphological and molecular phylogenetic investigations (Bailey et al. 2006; Materials and Methods Al-Shehbaz et al. 2006 ; Alexander et al. 2006 ). Taxon Sampling and DNA Isolation— Leaf material for phylogenetic Thysanocarpus is notable for its unusual fruits, which are analysis of Thysanocarpus was obtained from recent collections and from orbicular to obovate winged silicles, unilocular, one-seeded, herbarium specimens from the following institutions: CAS, DS, JEPS, MO, NMC, RSA, and UC (see Appendix 1). Specimens were chosen to and indehiscent. These fruits may be glabrous or pubescent include multiple samples of all taxa recognized in treatments of the genus with simple, blunt-tipped trichomes. The only other species by Rollins (1993) and Munz (1959) and to include as much of the mor- in the western United States with similar fruits is Athysanus phological variation within T. curvipes and T. laciniatus as possible. A total pusillus (Hook.) Greene, which differs from Thysanocarpus in of 55 specimens of Thysanocarpus were used, including 29 specimens of the following respects: 1) the silicles are pubescent with unci- T. curvipes; the geographic distribution of these samples is provided in Figs. 1 and 2 . DNA was isolated following the procedures of Alexander F1 F2 nate trichomes; 2) the silicles are borne in secund infructes- et al. (2007) , except that 10 m m Tris-HCl in 70% EtOH was used instead cences; and 3) the foliage is pubescent with branched of 70% EtOH when washing contaminants from the silica columns trichomes. Fruit similarities led A. pusillus to be named ini- at step 13. tially as a species of Thysanocarpus (Hooker 1836). However, a To assess the monophyly of Thysanocarpus and its placement within Brassicaceae, we selected 51 samples of the family from GenBank for careful reexamination of morphology (Al-Shehbaz et al. 2006) which both ITS and trnL-F spacer sequences were available. Sequences in conjunction with preliminary molecular phylogenetic anal- were chosen to sample broadly across the named tribes of Brassicaceae yses (Alexander et al. 2006; see also Results) suggests an affili- and include as many species of tribe Thelypodieae as possible (tribes fol- ation with members of Arabideae and no particularly close low Al-Shehbaz et al. 2006 , Al-Shehbaz and Warwick 2007 ; Warwick et al. relationship with Thysanocarpus . Outside of North America 2009 ) The complete list of accessions is provided in Appendix 1. Amplification and Sequencing—Two loci were amplified and there are several additional genera with fruits similar to those sequenced; trnL-F (primers F and C, Taberlet et al. 1991) from the chloro- of Thysanocarpus in the tribes Alysseae, Heliophileae, and plast genome and the nuclear ribosomal ITS (primers ITS5 and ITS4, White Isatideae. The similarity of Thysanocarpus to Tauscheria Fisch. et al. 1990). Amplification and sequencing followed previously developed (currently included in Isatis L.) in particular was noted by protocols ( Bailey and Doyle 1999 ; Bailey et al. 2002 ), and both loci were successfully amplified and sequenced for all samples except trnL-F for Hooker (1829), Torrey and Gray (1838–40), and Greene (1891). one accession of T. desertorum (32) and ITS for one T. curvipes (26) and one However, recent molecular phylogenetic investigations sug- T. erectus (38). Most sequences were generated by direct sequencing of PCR gest that these taxaProof with similar fruits are not closely related products.Only However, initial direct sequencing attempts for ITS products in 1 2 SYSTEMATIC BOTANY [Volume 35

Thelypodieae. The sequence data for T. laciniatus includes conflict between the two loci caused by hybridization (see Discussion). Consequently, application of cladistic methodology to a concatenated dataset including trnL-F and ITS sequences for T. laciniatus is inappropriate, because the resulting terminals would violate a fundamental assumption of cladistic analysis, that character state change occurs along a common bifurcat- ing evolutionary history ( Bremer and Wanntorp 1979 , Brower et al. 1996 , Proof CronquistOnly 1987 ). Matrix 1 was rooted with Aethionema grandiflorum Boiss. & Hohen., a member of the genus that is well-supported as sister to all other extant Brassicaceae ( Al-Shehbaz et al. 2006 ; Bailey et al. 2006 ). Matrices 2, 3, and 4 are used to investigate relationships within Thysanocarpus . Matrix 2 contains ITS sequences for all Thysanocarpus samples, including multiple sequences from accessions of T. laciniatus as described above. Matrix 3 contains trnL-F sequences for all Thysanocarpus samples. Matrix 4 is a concatenated data set containing both ITS and trnL- F samples for all Thysanocarpus except T. laciniatus. The single accession (26) of T. curvipes for which only a trnL-F sequence was available was also excluded from Matrix 4. Phylogenetic relationships among members of Thelypodieae are insufficiently resolved to identify the sister group of Thysanocarpus (Bailey et al. 2006; and see Results). To root our analyses, we chose Streptanthella longirostris (S. Wats.) Rydb., a member of Thelypodieae that is resolved in a large polytomy with Thysanocarpus . However, identical interspecific relationships were recovered in analyses including all sampled members of Thelypodieae (Matrix 1; Fig. 3 ; see Results) suggest- F3 ing that outgroup selection has not affected inferred relationships within Thysanocarpus . Matrices were aligned automatically using ClustalW ( Thompson et al. 1994) under default parameters. The resulting alignments were then checked and refined manually using Se-Al ( Rambaut 2002 ). Gap characters were included for each locus using the simple indel coding method of Simmons and Ochoterena (2000) as implemented in SeqState (Müller 2005). Aligned lengths and numbers of parsimony-informative nucleotide and gap characters for each matrix are indicated in Table 1. All matrices T1 are deposited in TreeBASE (study number S2546). For concatenated matrices including both loci (Matrix 1 and Matrix 4), incongruence length difference tests (Farris et al. 1995) as implemented in WINCLADA (Nixon 1999–2002) were conducted to determine if there is Fig. 1 . Map of the western U. S. A. and adjacent Mexico showing the significant incongruence between the loci. For each of these two matrices, geographic distribution of Thysanocarpus curvipes samples used in phylo- comparison of ITS and trnL-F found them not to be significantly incongru- genetic analyses. All maps were created in Quantum GIS (2008) . ent (Matrix 1, p = 0.0909; Matrix 4, p = 1.0000). Prior to parsimony analyses, uninformative characters were removed from the matrices. For each matrix, heuristic searches for most parsimoni- some T. laciniatus gave poor reads suggesting polymorphism among ITS ous trees were conducted using NONA (Goloboff 2000) spawned from copies in the genome. WINCLADA with 500 random addition sequence replicates with tree Polymorphism in ITS reads for accessions 41, 44, and 49 of bisection and reconnection (TBR), holding 20 trees per replicate, followed Thysanocarpus laciniatus was addressed by cloning of ITS PCR products by further TBR searching with a maximum of 10,001 trees. Clade support (using Promega TA cloning) and by a restriction digest approach. Cloning was estimated with 1,000 jackknife resampling replicates ( Farris et al. can isolate and amplify individual copies of ITS from within a single pool 1996 ) under default settings in TNT ( Goloboff et al. 2008 ) using the “new of PCR product, allowing multiple haplotype sequences to be recovered. technology search”, with parsimony ratchet (Nixon 1999), sectorial search, Cloning was followed by direct sequencing from 10 colonies for each tree-drifting, and tree-fusing (Goloboff 1999) techniques activated and a sample. Two divergent ITS sequence types were recovered for each of 36% removal probability. Additionally, maximum likelihood analyses these three samples. One sequence from each grouped with the “con- were conducted to test for sensitivity of the results to different methods of chuliferus clade” (see Results) and the other with T. curvipes . The single analysis. Maximum likelihood analyses were conducted with the online ITS sequences from other T. laciniatus samples similarly grouped either RAxML Black Box server ( Stamatakis et al. 2008 ), which uses the GTR + in the conchuliferus clade or with T. curvipes. This prompted attempts to Γ model when conducting a heuristic search and the GTR + CAT model isolate additional ITS sequence types from other samples of T. lacinia- when computing bootstraps. The matrices analyzed by maximum likeli- tus using a restriction enzyme approach ( Hughes et al. 2002 ). For each hood were identical to those included in the parsimony analyses described of these samples one ITS sequence type had already been recovered, so above except that gaps were excluded. We chose this approach because it restriction enzymes were chosen to selectively cut the known ITS type is not clear whether the models used by RAxML, or any other available from a sample of genomic DNA, allowing any divergent copies of ITS to models, are appropriate for modelling the evolution of gap characters. For be amplified and sequenced. For all T. laciniatus samples for which only the concatenated Matrices 1 and 4, model parameters were estimated for a sequence from the conchuliferus clade had been recovered, 1.1 μL of a 9:1 each locus separately. mixture of buffer NEB4 and Hpy188I (enzymes and buffers from New England BioLabs) was incubated with 9 μL of genomic DNA at 37°C for μ two hours. If only a T. curvipes sequence had been recovered, 2.1 L of a Results 10:10:1 mixture of buffer NEB3, bovine serum albumen, and CviQI was incubated with 8 μL of genomic DNA at 25°C for two hours. The restric- Monophyly and Phylogenetic Placement of Thysanocarpus— tion digest products were then used as a template for ITS amplification Figure 3 shows the cladogram resulting from phyloge- and sequencing. This resulted in the recovery of additional divergent ITS sequence types for samples 46 and 48. This restriction enzyme approach netic analysis of Matrix 1. Thysanocarpus is placed within failed to yield additional ITS sequence types for T. laciniatus accessions Thelypodieae, which is found to be monophyletic with strong 39, 40, 42, and 45. (87% jackknife) support. Resolution within Thelypodieae, Alignment, Gap-coding, and Phylogenetic Analysis— Four separate however, is limited, with Thysanocarpus placed in a large matrices were analyzed using parsimony. Matrix 1, intended to test the basal polytomy. Thysanocarpus is weakly supported (< 50%) monophyly and placement of Thysanocarpus , contains both ITS and trnL- F sequences and includes accessions of six species of Thysanocarpus (all as monophyletic; this low support is associated primarily except T. laciniatus ) withinProof a broad sampling across Brassicaceae and withOnly the placement of T. erectus , as the remaining species are 2010] ALEXANDER ET AL.: THYSANOCARPUS PHYLOGENETICS AND TAXONOMY 3 Proof Only

Fig. 2 . Geographic distribution of additional Thysanocarpus specimens used in phylogenetic analyses. A. Thysanocarpus conchuliferus, T. desertorum, T. erectus, and T. rigidus samples from southern California and Baja California in the U. S. A. and Mexico, respectively. B. Thysanocarpus laciniatus and T. radians samples from California and Nevada, U. S. A.

monophyletic with high (94%) support. Athysanus pusillus is the trnL-F analysis, (Fig. 5) but this grouping receives strong strongly supported (100%) as sister to Draba nemorosa L. of support (> 95%) in the ITS and combined analyses ( Figs. 4 , 6 ). Arabideae. A “northwestern Nevada clade”, including three specimens F4 Relationships Among Species of Thysanocarpus—Figures 4 , of T. curvipes subsp. curvipes from Humboldt and Washoe F5 F6 5 , and 6 show cladograms resulting from phylogenetic anal- Cos. also is apparent in all three analyses, receiving weak yses of Matrices 2, 3, and 4, respectively. All three analyses support (64%) in the ITS analysis and stronger support show T. erectus sister to the remainder of the genus, in agree- (> 85%) in the trnL-F and combined analyses. A third sub- ment with Fig. 3 . The other species of the genus are divided clade, comprising all sampled specimens of T. curvipes subsp. into two clades, one including all samples of T. conchulif- elegans, is strongly supported (> 95%) in the ITS and com- erus, T. desertorum , and T. rigidus (the “conchuliferus clade”) bined analysis (Figs. 4, 6 ) but these accessions are unresolved and the other including all accessions of T. curvipes and in the trnL-F analysis ( Fig. 5 ). Additional resolution within T. radians. The only hard incongruence among these analy- T. curvipes is found in ITS and combined analyses, but is only ses involves the placement of T. laciniatus. In the plastid data weakly supported (< 65%). set ( Fig. 5 ), all accessions of T. laciniatus are in the curvipes clade, either forming a clade sister to T. curvipes or, in the Discussion case of Accession 49, in a polytomy including all accessions of that species. In the nuclear ITS data set (Fig. 4), acces- Monophyly and Phylogenetic Placement of Thysanocarpus— sions of T. laciniatus fall in two distinct clades. “Laciniatus Thysanocarpus is placed in a large basal polytomy within clade 1” is strongly supported (99% jackknife) as a mem- Thelypodieae, consistent with earlier phylogenetic analysis ber of the conchuliferus clade whereas “laciniatus clade 2” is based solely on ITS sequence data (Bailey et al. 2006). The strongly supported (97%) as a member of the curvipes clade. tribe appears to be recalcitrant to phylogenetic analysis, as no ITS sequences belonging to both of these clades (denoted by published analysis has yielded significant resolution within suffixes “A” and “B” after accession numbers) were recov- it (Bailey et al. 2006; Beilstein et al. 2006; Beilstein et al. 2008). ered from five accessions (41, 44, 46, 48, and 49) of T. lacin- More precise knowledge of the close relatives of Thysanocarpus iatus, three of which are known polyploids (Windham et al. must therefore await further investigation. Support for mono- unpubl.). phyly of Thysanocarpus exclusive of T. erectus is high, but Phylogenetic Resolution within Thysanocarpus curvipes— T. erectus is only weakly united with the remainder of the Resolution within T. curvipes is low, however two subclades genus. This is not surprising given the poor resolution at the of T. curvipes are found in all three analyses. The only two genus level throughout Thelypodieae. Placement of Athysanus specimens of T. curvipes subsp. longistylus included in our in Arabideae is confirmed, consistent with earlier morpho- sample form a weaklyProof supported (64% jackknife) subclade in logicalOnly ( Al-Shehbaz et al. 2006 ) and phylogenetic ( Alexander 4 SYSTEMATIC BOTANY [Volume 35 Proof Only

Fig. 3 . Strict consensus of 10,001 most parsimonious trees (length = 1,740, CI = 0.37, RI = 0.48) from parsimony analysis of ITS and trnL-F sequences for broad sample of Brassicaceae including 6 species of Thysanocarpus . Numbers above branches indicate parsimony jackknife support, while numbers below branches indicate maximum likelihood bootstrap support. Support values below 50 are not shown, and a “–“ indicates that the node in question did not occur on the best tree from maximum likelihood analysis. et al. 2006) analyses. Similarity between fruits of the two gen- Relationships Among Species of Thysanocarpus—The era is thus due to convergence, in agreement with recent sug- Baja California endemic T. erectus forms the earliest diverg- gestions that fruit morphology is a poor guide to relationships ing branch in all analyses (Figs. 3–6) and is sister to the other within the family as a whole ( Beilstein et al. 2006 ; Beilstein species of Thysanocarpus. The remaining species are divided et al. 2008 ). Proof intoOnly two clades, one including all samples of T. conchuliferus , 2010] ALEXANDER ET AL.: THYSANOCARPUS PHYLOGENETICS AND TAXONOMY 5

Table 1. Aligned lengths, numbers of parsimony-informative charac- 46, and 48) contain one ITS sequence type derived from the ters, and numbers of parsimony-informative gap characters for each locus conchuliferus clade (“laciniatus clade 1”), and a second type in each matrix. derived from T. curvipes (“laciniatus clade 2”). Two accessions,

ITS trnL-F 47 and 49, show different patterns. Although one ITS sequence type from the conchuliferus clade and one from the curvipes parsimony- parsimony- parsimony- parsimony- aligned informative informative aligned informative informative clade were recovered from Accession 49, these sequence types length charactersProofgap characters length characters gap characters Only do not group with either laciniatus clade 1 or laciniatus clade 2. Matrix 1 600 211 29 1,284 133 40 The two ITS sequence types recovered from Accession 47 are Matrix 2 689 103 9 both derived from the conchuliferus clade, one within lacinia- Matrix 3 697 25 6 tus T. desertorum Matrix 4 689 96 9 697 23 5 clade 1 and the second associated with . These results provide strong evidence that nearly all individuals of T. laciniatus included in our sample represent hybrids between the conchuliferus clade and the curvipes clade, T. desertorum, and T. rigidus (the conchuliferus clade) and the and that T. laciniatus has arisen more than once, potentially as other including all accessions of T. curvipes and T. radians many as three times among the specimens examined in the (Fig. 6). Each member of the conchuliferus clade has, at one present study. Further, the hybridization event giving rise to time or another, been treated as a variety or synonym of the lineage represented by Accessions 41, 44, 46, and 48 may T. laciniatus (Munz 1959; Rollins 1993). Although T. conchuliferus have occurred after the more rapid coalescence of organel- was considered a distinct species in the most recent synopsis of lar loci but before the slower coalescence of nuclear loci that the genus ( Rollins 1993 ), T. desertorum (occasionally recognized results from the four-fold greater effective population size as T. laciniatus var. hitchcockii Munz) and T. rigidus (occasionally of the latter (Birky et al. 1983). This would explain the recip- recognized as T. laciniatus var. rigidus Munz) have rarely been rocal monophyly of T. curvipes and this lineage of T. lacinia- recognized except by the most avid splitters. However, the dis- tus in analysis of trnL-F sequences ( Fig. 5 ), but the absence covery of the hybrid origin of typical T. laciniatus (see below) of reciprocal monophyly between laciniatus clade 2 and requires a reassessment of past taxonomic treatments. If the T. curvipes in analysis of ITS sequences ( Fig. 4 ). However, the Santa Cruz Island endemic T. conchuliferus is to be maintained apparent lack of coalescence of ITS sequences for T. curvipes as a species, as has become customary in recent years ( Rollins could also be confounded by recombination or a lack of suffi- 1993 ), then the equally distinct T. desertorum and T. rigidus cient informative variation at this level of analysis. Additional must be afforded the same treatment. All three of these taxa chromosome counts and a more detailed examination of the are diploids (Windham et al. unpubl.) and subsuming them origins of T. laciniatus are ongoing to better understand both under T. laciniatus (a name that clearly applies to a hybrid) is the complex, multiple origins of T. laciniatus and cytological both unhelpful as a representation of the evolutionary relation- evolution in the genus as a whole. Similar complicated his- ships of these taxa and disallowed under Article H.5.1 of the tories have been documented in a variety of allopolyploid International Code of Botanical Nomenclature ( McNeill et al. taxa ( Werth et al. 1985 ; Wyatt et al. 1988 ; Soltis and Soltis 2006). Thysanocarpus conchuliferus, T. desertorum, and T. rigidus 1999; Rauscher et al. 2004), and a more complete exposi- are rather easily separated from T. laciniatus (and each other) tion of the evolution of T. laciniatus will be the subject of a and are treated below as distinct species. future paper. Hybrid Origin of Thysanocarpus laciniatus— A hybrid ori- Phylogenetic Resolution within Thysanocarpus curvipes— gin for T. laciniatus is indicated by two related lines of evi- There is strong support for monophyly of T. curvipes subsp. dence: 1) differing placement of some sequences from this elegans from ITS ( Fig. 4 ) but not from trnL-F ( Fig. 5 ). The lack of species in cladograms inferred from plastid and nuclear support for this group in trnL-F data apparently results from sequence data, and 2) polymorphism among copies of ribo- the lower variabiliy of trnL-F compared to ITS (see Table 1 ). somal DNA within individuals of T. laciniatus . Ten out of Assessment of herbarium specimens suggests that subsp. eleven samples of T. laciniatus exhibit incongruence between elegans introgresses regularly with other infraspecific taxa. analyses based on trnL-F and ITS sequences. In the trnL-F Although subsp. elegans is readily recognizable in its extreme cladogram (Fig. 5), all accessions of T. laciniatus are strongly form, a substantial number of herbarium specimens are mor- supported as members of the curvipes clade (93% jackknife phologically intermediate between this subspecies and subsp. support). This placement is in conflict with the ITS cladogram curvipes . It nonetheless is distinct at one of the two sampled ( Fig. 4 ), which indicates that all accessions but one (Accession loci, is generally recognizable morphologically, and is here 42) of T. laciniatus contain an ITS sequence type derived from given subspecific status. the conchuliferus clade, most of these forming laciniatus clade 1. The T. curvipes subsp. longistylus clade appears in all anal- For Accession 42, the only specimen lacking conflict between yses with strong support. Subspecies longistylus is poorly ITS and trn L-F, both loci indicate membership in the curvipes represented in the current sequence data, with only two spec- clade. Given the similarity between the sequences recovered imens. Although this taxon is here given subspecific status, from Accession 42 and those obtained from several putative more extensive sampling of individuals and loci would be allopolyploids (Accessions 41, 44, and 46), it is possible that required to establish with greater certainty whether this taxon this specimen lost the ITS sequence representing the conchu- is genetically distinct from other T. curvipes . liferus clade through concerted evolution (e.g., Wendel et al. The northwestern Nevada clade of T. curvipes provides the 1995 ; Rauscher et al. 2004 ). only strongly-supported phylogenetic resolution in the spe- Polymorphism among nuclear ribosomal ITS copies was cies that has no apparent correlation with morphology. These detected in six accessions of T. laciniatus ( Fig. 4 ), providing are typical members of subsp. curvipes , essentially indis- further evidence for a hybrid origin of this taxon. Cloning and tinguishable from plants occurring in northern California, restriction site analysesProof revealed that four accessions (41, 44, Idaho,Only Oregon, and Washington. None of the other members 6 SYSTEMATIC BOTANY [Volume 35 Proof Only

Fig. 4 . Strict consensus of 54 most parsimonious trees (length = 176, CI = 0.75, RI = 0.96) from parsimony analysis of ITS sequences of Thysanocarpus , rooted with Streptanthella longirostris. Numbers above branches indicate parsimony jackknife support, while numbers below branches indicate maximum likelihood bootstrap support. Support values below 50 are not shown, and a “–“ indicates that the node in question did not occur on the best tree from maximum likelihood analysis.Proof Accessions of T. laciniatus falling outside of laciniatus Only clade 1 and laciniatus clade 2 are marked with asterisks. 2010] ALEXANDER ET AL.: THYSANOCARPUS PHYLOGENETICS AND TAXONOMY 7 Proof Only

Fig. 5 . Single most parsimonious tree (length = 35, CI = 0.94, RI = 0.98) from parsimony analysis of trnL-F sequences of Thysanocarpus , rooted with Streptanthella longirostris. Numbers above branches indicate parsimony jackknife support, while numbers below branches indicate maximum likelihood bootstrap support. Support values below 50 are not shown, and a “–“ indicates that the node in question did not occur on the best tree from maximum likelihood analysis. An accession of T. laciniatus falling outside the main T. laciniatus clade is marked with an asterisk. Note also that incongruence between parsimony and maximum likelihood analyses regarding the monophyly of the two samples of T. desertorum apparently results from the inclusion of gap characters in the former analysis but not the latter. Gap characters provide two synapomorphis joining these two samples, but in the absence of gaps there are no synapomorphiesProof for T. desertorum . Only 8 SYSTEMATIC BOTANY [Volume 35 Proof Only

Fig. 6 . Strict consensus of 22 most parsimonious trees (length = 180, CI = 0.83, RI = 0.96) from parsimony analysis of combined ITS and trnL-F sequences of Thysanocarpus (excluding T. laciniatus ), rooted with Streptanthella longirostris . Numbers above branches indicate parsimony jackknife support, while numbers below branches indicate maximum likelihood bootstrap support. Support values below 50 are not shown, and a “–“ indicates that the node in question did not occur on the best tree from maximum likelihood analysis.

of subsp. curvipes (accessions 15, 16, 25 and 31) is resolved branched. Basal leaves shortly petiolate, simple, entire or den- with these plants in any analysis. The genetic distinctness of tate to pinnatifid or pinnatisect, often withered by anthesis or the northwestern Nevada clade most likely reflects histori- fruiting; cauline leaves sessile, auriculate or not, entire or den- cal isolation that has not been accompanied by any readily tate or pinnatifid to pinnatisect. Racemes several, many flow- apparent morphological divergence. ered, corymbose, ebracteate, usually elongated considerably in fruit; fruiting pedicels slender, often recurved. Sepals green- Taxonomic Treatment ish or purplish with pale margins, oblong to ovate, ascending, caducous, base of lateral pair not saccate; petals white to Thysanocarpus Hook., Fl. Bor.-Amer. 1: 69–70. 1829.—TYPE: purplish, 1–3 mm long, subequalling or longer than sepals, Thysanocarpus curvipes Hook. blade spatulate to oblong, claw undifferentiated from blade; Herbs, annual, glaucous, glabrous or pubescent. Trichomes stamens 6, slightly tetradynamous; filaments slightly dilated of the foliage absent or concentrated towards the base of the at base; anthers ovate, yellow or purple; lateral nectaries 1 on plant, simple, acicular,Proof 0.3–0.6 mm long. Stems erect, simple or eachOnly side of lateral stamens or semiannular, median nectaries 2010] ALEXANDER ET AL.: THYSANOCARPUS PHYLOGENETICS AND TAXONOMY 9

absent. Fruits indehiscent silicles 2.5–10 mm wide, orbicular ate, often perforate; septum obsolete; ovules and seeds 1 per to ovate or elliptic, strongly latiseptate, sessile, unsegmented, fruit; style short, 0.2–1.5(–1.8) mm long, usually persistent but usually pendulous; valves with a prominent midvein, gla- fragile and easily broken; stigma entire. Seeds aseriate, wing- brous or with blunt-tipped, simple trichomes; replum winged, less, elliptic to orbicular, flattened; seed coat not mucilaginous the wing flattened, Proofusually with radiating rays, entire or cren- whenOnly wetted; cotyledons accumbent. 1. Fertile anthers purple (flowers occasionally with whitish or yellowish infertile anthers) ...... 2 2. Middle cauline leaves lanceolate, widest near the base, bases auriculate-clasping, auriculae extending around the stem in at least some leaves; basal leaves subentire to sinuate-dentate ...... 2. Thysanocarpus curvipes 2. Middle cauline leaves linear, widest near the middle or equally wide throughout, bases cuneate or inconspicuously auriculate; auriculae, when present, never extending around the stem; basal leaves subentire to pinnatifid ...... 3 3. Pedicels straight or nearly so, horizontal to ascending; plants branching from the base; foliage purplish; fruits often purplish as well ...... 7. Thysanocarpus rigidus 3. Pedicels recurved in a continuous arc; plants usually not branching from the base, foliage greenish throughout or purplish in the basal rosette only; fruits greenish ...... 5. Thysanocarpus laciniatus 1. Fertile anthers yellow ...... 4 4. Fruits cymbiform, strongly incurved toward the flat side of the silicle, the wings radiate and divided into spatulate lobes or occasionally perforate ...... 1. Thysanocapus conchuliferus 4. Fruits flat or plano-convex, the wings radiate or eradiate, entire to shallowly incised, not incurved toward the flat side of the silicle ...... 5 5. Lower fruiting pedicels 7–18 mm long, the tips geniculately reflexed; fruits 7–10 mm wide, the wings with distinct rays ± 0.1 mm wide ...... 6. Thysanocarpus radians 5. Lower fruiting pedicels 2–6 mm long, recurved in a continuous arc or straight; fruits 2.5–5.5 mm wide, the wings with indistinct rays 0.2–0.5 mm wide or eradiate ...... 6 6. Fruiting pedicels erect to ascending proximally, often smoothly recurved distally; fruits pubescent with blunt hairs 0.2–0.3 mm long or occasionally glabrous; wings of the fruit eradiate and deeply emarginate at base ...... 4. Thysanocarpus erectus 6. Fruiting pedicels stiffly spreading, straight or nearly so; fruits usually pubescent with minute clavate hairs 0.05–0.1 mm long, occasionally glabrous; wings of the fruit radiate or not, broadly cuneate or truncate at base, never deeply emarginate ...... 3. Thysanocarpus desertorum

1. Thysanocarpus conchuliferus Greene, Bull. Torrey 2. Thysanocarpus curvipes Hook., Fl. Bor.-Amer. 1: 69–70. Bot. Club 13: 218. 1886. Thysanocarpus laciniatus Nutt. 1829.—TYPE : U. S. A.: Near the boundary between ex Torr. & A. Gray var. conchuliferus (Greene) Jeps. Man. Wasco Co., Oregon, and Klickitat Co., Washington: On Fl. Pl. Calif. 447. 1925.—TYPE : U. S. A. California: Santa moist ground, near the Great Falls of the Columbia, s. d., Barbara Co.: Island of Santa Cruz, July and August 1886, D. Douglas s. n. (holotype: K!; isotype: GH). E. L. Greene s. n. (holotype: NDG!). Thysanocarpus runcinatus G. Don, A General History of the Thysanocarpus conchuliferus Greene var. planiusculus B. L. Rob., Dichlamydeous Plants, 1: 196. 1831.—LECTOTYPE , here Syn. Fl. N. Amer. 1: 113. 1896.—TYPE : U. S. A. California: designated: Table 18 A, W. J. Hooker, Fl. Bor.-Amer. 1: Santa Barbara Co.: Santa Cruz Island, April 1888, T. S. 388. 1829. Although the specimen used in producing this Brandegee s. n. (holotype: GH; isotypes: GH, UC!). figure was not explicitly stated, it was likely drawn from Trichomes absent. Basal leaves oblanceolate to elliptic, the holotype of T. curvipes . pinnatifid or occasionally sinuate-dentate, 1–3(–5) cm long. Trichomes present or absent. Basal leaves oblanceolate Cauline leaves lanceolate to narrowly elliptic or nearly lin- to obovate, subentire to sinuate-dentate, 1–6(–13) cm long. ear, widest near base or middle, auriculate-clasping. Flowers Cauline leaves lanceolate, widest at base, auriculate-clasping. with yellow anthers. Lower fruiting pedicels 3.5–6.5 mm Flowers with purple anthers, or rarely some flowers with long, stiffly divaricate-ascending or slightly recurved. Fruits infertile, whitish anthers. Lower fruiting pedicels 3–7(–12) glabrous, cymbiform, the wings strongly incurved toward the mm, recurved in a continuous arc. Fruits with clavate hairs flat side of the silicle; wings divided into spatulate lobes 0.25– 0.2–0.4 mm long or glabrous, flat or plano-convex, obovate 0.4 mm wide at the narrowest, these occasionally joined dis- to nearly orbicular, 3–6(–9) mm wide; wing entire, crenate, F7 tally. Figure 7A . divided into narrow, spatulate lobes, or perforate, usually Distribution—United States, southern California, Santa with broad (0.2–0.5 mm) rays ( Fig. 9 ). F9 F8 Cruz Island. Figure 8 . Distribution—United States (Arizona, California, Colorado, Thysanocarpus conchuliferus is the most narrowly-distributed Idaho, Montana, Nevada, New Mexico, Oregon, Utah, and species in the genus, found only on Santa Cruz Island. The Washington), Mexico (northern Baja California and north- cymbiform fruits of the species are very distinctive, although ern Sonora), and Canada (southern Vancouver Island, British T. curvipes occasionally has silicles with incurved to invo- Columbia); Fig. 10 . F10 lute wings. In addition to characters used in the key, the two This is a variable species and many attempts have been are further distinguished by the much shorter, denser inflo- made to divide that variation into recognizable taxa. These rescences of T. conchuliferus (internodes in the inflorescence efforts have produced a total of 18 basionyms referrable to average less than 2 mm in T. conchuliferus and greater than plants here included within T. curvipes . Morphological charac- 3 mm in T. curvipes ). Although T. conchuliferus has usually ters used include pubescence of the stem, foliage, and silicle, been considered a variety of T. laciniatus in the literature, it length of the style, and size or ornamentation of the sili- has strongly auriculate cauline leaves, while T. laciniatus has cle wing. Of these characters, all but pubescence of the cuneate-based or minutelyProof auriculate leaves. silicleOnly show variation along geographic lines. Pubescent 10 SYSTEMATIC BOTANY [Volume 35 Proof Only

Fig. 7 . Fruit morphology of Thysanocarpus species except T. curvipes . A. T. conchuliferus , U. S. A.: California: Santa Barbara Co., 14 Apr 1985, S. A. Junak SC-1053 (RSA). B. T. desertorum, isotype, U. S. A.: California: Kern Co., 14 Apr 1905, A. A. Heller 9701 (POM). C. T. erectus, Mexico: Baja California: Mcpio. Ensenada, 4 Apr 1991, S. Boyd 5756 (RSA). D. T. laciniatus , U. S. A.: California: San Diego Co., 24 Apr 2006, M. D. Windham 3424 (NMC). E. T. radians , U. S. A.: California: Butte Co., 13 Apr 1986, B. Ertter 5884 (RSA). F. T. rigidus , holotype, U. S. A.: California: San Diego Co., 16 Mar 1925, P. A. Munz 9701 (POM).

silicles appear to be more common in the northern part of in some areas of southern California. Two subspecies, subsp. the range of T. curvipes but this feature is variable and, con- elegans and subsp. longistylus , occur only within the ranges trary to Piper’s (1906) suggestion that fruit pubescence is con- of other subspecies of T. curvipes . The latter two subspecies sistent within populations, we have seen a number of mixed receive some support in our molecular phylogenetic analyses, populations. while the previous three do not. Although the five subspecies Based on concordance between morphology and geog- recognized here are quite distinct in their extremes, there are a raphy, with insight added by phylogenetic analyses as dis- number of specimens of intermediate morphology, as is to be cussed above, we recognize five subspecies within T. curvipes . expected for infraspecific taxa. Southern California is particu- Three of these, subsp. amplectens , subsp. curvipes , and subsp. larly problematic, as all five subspecies overlap in this area eradiatus , are predominantlyProof allopatric, although overlapping andOnly all appear to interbreed to some extent. 2010] ALEXANDER ET AL.: THYSANOCARPUS PHYLOGENETICS AND TAXONOMY 11 Proof Only

Fig . 8. Distribution of other Thysanocarpus in the western U. S. A. and Baja California in adjacent Mexico. D istribution is mapped at the county/municipio level except for T. rigidus and T. erectus , for which all specimens seen are mapped. Proof Only 12 SYSTEMATIC BOTANY [Volume 35 Proof Only

Fig. 9 . Fruit morphology of Thysanocarpus curvipes subspecies. A. T. curvipes subsp. amplectens , Mexico: Sonora: Mcpio. Tubutama, 2 Apr 2005, T. R. Van Devender 2005–479 (NMC;). B. T. curvipes subsp. curvipes, U. S. A.: Nevada: Washoe Co., 12 May 1974, M. J. Williams 74–190–7 (RSA). C. T. curvipes subsp. elegans , U. S. A.: California: Butte Co., 8 May 2006, L. Ahart 12608 (RSA). D. T. curvipes subsp. eradiatus , U. S. A.: California: Imperial Co., 27 Feb 2005, M. D. Windham 3295 (NMC). E. T. curvipes subsp. longistylus , U. S. A.: California: Tulare Co., 12 May 1949, P. A. Munz 13178 (RSA).

1. Stems pubescent basally ...... 2 2. Fruits < 5.5 mm wide and 6 mm long, entire, incised, or perforate ...... 3 3. Styles ≤ 0.7 mm long ...... 2b. subsp. curvipes 3. Styles 0.9–1.8 mm long ...... 2e. subsp. longistylus 2. Fruits ≥ 5.5 mm wide and 6 mm long, perforate ...... 2c. subsp. elegans 1. Stems glabrous throughout ...... 4 4. Styles 0.9–1.8 mm long ...... 2e. subsp. longistylus 4. Styles ≤ 0.7 mm long ...... 5 5. Wings of the fruit radiate, entire to perforate ...... 2a. subsp. amplectens 5. Wings of theProof fruit eradiate, entire or sparingly incised ...... Only...... 2d. subsp. eradiatus 2010] ALEXANDER ET AL.: THYSANOCARPUS PHYLOGENETICS AND TAXONOMY 13 Proof Only

Fig. 10 . Map of western North America showing the distribution of the subspecies of Thysanocarpus curvipes. Distribution is mapped at the county/ municipio level except for Canada, which lacks equivalent political units. Proof Only 14 SYSTEMATIC BOTANY [Volume 35

2 a. Thysanocarpus curvipes Hook. subsp. amplectens (Greene) Trichomes present. Fruits 3–5.5 mm wide, the wings radi- P. J. Alexander & Windham, comb. nov. Thysanocarpus ate, entire, crenate, divided into spatulate lobes, or perforate, amplectens Greene, Pittonia 3: 87. 1896.—TYPE : U. S. A. the style 0.2–0.6(–7) mm long. Figure 9B . New Mexico: Grant: Duck Creek Hills, 16 Apr 1880, E. L. Distribution— United States (California, Idaho, southwest- Greene s. n. (holotype: NDG!). ern Montana, northern Nevada, Oregon, and Washington), Thysanocarpus affinisProof Greene, Pittonia 4: 311–312. 1901.—TYPE: MexicoOnly (northern Baja California and northern Sonora), U. S. A. California: Los Angeles Co.: Santa Catalina Island, and Canada (southern Vancouver Island, British Columbia). Avalon, March 1901, B. Trask s. n. (holotype: NDG!; iso- Figure 10 . types: NY!, US!). This is the most widespread subspecies of Thysanocarpus curvipes and further shows the greatest sympatry with other Thysanocarpus filipes Greene, Pittonia 4: 200–201. 1900.—TYPE : subspecies. The ranges of both subsp. elegans and subsp. U. S. A. Arizona: Greenlee Co.: Coronado Rim near Clifton, longistylus are included almost entirely within the distribu- 1899, A. Davidson s. n. (holotype: NDG!, isotypes: POM!). tion of subsp. curvipes. Stems of these three subspecies are Thysanocarpus ramosus Greene, Bull. Calif. Acad. Sci. 2: 390. pubescent basally (with rare exceptions in subsp. longistylus ), 1887.—TYPE: U. S. A. California: Santa Barbara Co.: Island distinguishing them from the two southeastern subspecies, of Santa Cruz, July and August 1886, E. L. Greene s. n. subsp. amplectens and subsp. eradiatus . Specimens interme- (holotype: NDG!). diate between subsp. curvipes and subsp. elegans are encoun- Trichomes absent. Fruits 3.5–5.5(–6.5) mm wide, the wings tered throughout areas of sympatry, and these often cannot be radiate, entire, crenate, divided into spatulate lobes, or perfo- assigned confidently to either subspecies. Plants morphologi- rate, the style 0.2–0.7 mm long. Figure 9A . cally intermediate between subsp. curvipes and subsp. longi- Distribution— United States (southern California, southern stylus are, however, rare in herbarium collections. Arizona, and southwestern New Mexico) and Mexico (north- Variation within populations of T. curvipes subsp. curvipes ern Sonora). Figure 10 . can be substantial. For instance, silicles may be either gla- This and Thysanocarpus curvipes subsp. eradiatus are glabrous or pubescent, and either perforate or imperforate, at brous-stemmed plants found in the southeastern part of the a single site. As here defined, subsp. curvipes includes: var. range of T. curvipes. The two intergrade where their ranges cognatus (plants with cuneate-based silicles), subsp. madocar- overlap in southern California and central Arizona but are pus (plants with glabrous silicles), var. involutus (plants with otherwise distinct. The names T. amplectens and T. filipes strongly involute wings on the silicles), T. hirtellus (plants denote plants with glabrous, imperforate-winged silicles and with particularly abundant pubescence), and T. pulchellus those with pubescent, perforate-winged silicles, respectively, (plants with petals longer than the sepals). None of these variants that do not appear to warrant recognition. characters, however, seem to provide a sound basis for distin- 2 b. Thysanocarpus curvipes Hook. subsp. curvipes guishing taxa. Thysanocarpus crenatus usually has been listed as a synonym of T. laciniatus (e.g., Rollins 1993 ), but the type Thysanocarpus crenatus Nutt. ex Torr. & A. Gray, Fl. N. Amer. 1: material at BM and NY has prominently auriculate leaves and 118. 1829.—TYPE: U. S. A. California: Santa Barbara Co.: pubescent stems, closely resembling plants here assigned to Santa Barbara, s. d., T. Nuttall s. n. (holotype: BM!, iso- T. curvipes subsp. curvipes . type: NY!). 2 c. Thysanocarpus curvipes Hook. subsp. elegans ( Fisch. Thysanocarpus pulchellus Fisch. & C. A. Mey., Index Seminum & C. A. Mey.) P. J. Alexander & Windham, comb. nov. [St. Petersburg], 2: 50–51. 1835; Thysanocarpus curvipes Thysanocarpus elegans Fisch. & C. A. Mey., Index Seminum Hook. var. pulchellus (Fisch. & C. A. Mey.) Greene, Fl. [St. Petersburg] 2: 51. 1835; Thysanocarpus curvipes Hook. Francisc. 276. 1891.—TYPE: U. S. A. California: Sonoma: var. elegans (Fisch. & C. A. Mey.) B. L. Rob., Syn. Fl. N. Amer. Circa coloniam ruthenorum Ross, cult. in horto bot. 1(1): 114. 1895.—TYPE : U. S. A. California: Sonoma: prope imp. Petropol. [cultivated at the St. Petersburg Botanical coloniam Ross, 1834, F. P. Wrangel s. n. (holotype: LE!). Garden from material collected near the Russian Fort Ross], 1835, F. P. Wrangel s. n. (holotype: LE!). Thysanocarpus deppii Nutt. ex Torr. & A. Gray, Fl. N. Amer. 1: 118. 1838.—TYPE: U. S. A. California: s. d., Hooker s. n., Thysanocarpus curvipes Hook. var. involutus Greene, Fl. (holotype: K!). Francisc. 276. 1891.—TYPE: U. S. A. California: Sonoma Co.: 10 Mar 891, Bioletti s. n. (holotype: NDG!). Thysanocarpus foliosus Heller, Muhlenbergia 2: 47–48. 1905.— TYPE: U. S. A. California: Kern Co.: On the side of a ravine Thysanocarpus hirtellus Greene, Pittonia 3: 86. 1896.—TYPE: back of Girard Station in the Tehachapi Mountains, 18 U. S. A. California: Napa: Wooded cañon tributary to Dry Apr 1905, A. A. Heller 7719 (holotype: CAS!; isotypes: AC, Creek, 12 May 1895, Greene s. n. (lectotype, here designated: BKL, DS, GH, NY!, UC!, US!). NDG 4137!; isolectotypes: NDG 4136!, NDG 4138!). Trichomes present. Fruits 5.5–7(–9) mm wide, the wings Thysanocarpus curvipes Hook. subsp. madocarpus Piper, Cont. radiate, perforate, the style 0.4–0.8(–1) mm long. Figure 9C . U.S. Natl. Herb. 11: 306. 1906. Thysanocarpus curvipes Distribution—United States (California) and Mexico Hook. f. madocarpus J. F. Macbr., Cont. Gray Herb. 49: (northern Baja California). Figure 10 . 48. 1917.—TYPE : U. S. A. Washington: Whitman Co.: As mentioned above, this subspecies appears to hybrid- Pullman, May 1897, A. D. E. Elmer 79 (holotype: US!, iso- ize with Thysanocarpus curvipes subsp. curvipes on a regular types: BKL, NY!, POM!, UC!). basis. Plants that may result from hybridization with subsp. Thysanocarpus curvipes Hook. var. cognatus Jepson, Fl. Calif. amplectens or subsp. eradiatus can be found in the southern 2: 100. 1936.—TYPE : U. S. A. California: Tuolumne: Pine part of the range from Kern County to Santa Barbara County Log, South Fork Stanislaus River, 7 May 1916, A. L. Grant in southern California. Plants in the type collection of T. folio- 702 (holotype: ProofJEPS!). susOnly resemble these putative hybrids and, although tentatively 2010] ALEXANDER ET AL.: THYSANOCARPUS PHYLOGENETICS AND TAXONOMY 15 included here, are not readily assignable to any one subspe- which both ITS and trnL-F sequences form monophyletic cies. Morphological intermediates involving subsp. elegans are clades, but our sampling for this subspecies is limited and responsible for most of the apparent intergradation among none of the potential hybrids with subsp. eradiatus were subspecies of T. curvipes . Our molecular phylogenetic anal- included. yses provide support for the recognition of this subspecies, 3. Thysanocarpus desertorum Heller, Muhlenbergia 2: 47. although it is resolvedProof as a clade only in the ITS data. Only1905.—TYPE: U. S. A. California: Kern Co.: on rocky hill- Among plants here referred to Thysanocarpus curvipes tops near Randsburg, 14 Apr 1905, A. A. Heller 7681 (holo- subsp. elegans, the name T. deppii refers to plants with pubes- type: CAS!; isotypes: AC, BKL, DS, GH, MSC, NY!, US!). cent silicles. As discussed above, silicle pubescence shows a high degree of local variation independent of other characters Thysanocarpus laciniatus Nutt. ex Torr. & A. Gray var. hitch- and does not appear to be useful in naming taxa. cockii Munz, Bull. S. Calif. Acad. Sci. 31: 62. 1932.—TYPE: 2 d. Thysanocarpus curvipes Hook. subsp. eradiatus (Jepson) U. S. A. California: Inyo Co.: Dante’s Point, Death Valley, P. J. Alexander & Windham, comb. nov. Thysanocarpus 6 Apr 1928, P. A. Munz & C. L. Hitchcock 11016 (holotype: curvipes Hook. var. eradiatus Jepson, Man. Fl. Pl. Calif. POM!). 447. 1925.—TYPE: U. S. A. California: Inyo: Hanaupah Trichomes absent or, rarely, plants sparsely hirsute at base. Canyon, Panamint Mountains, 15 May 1917, W. L. Jepson Foliage greenish to purplish throughout. Basal leaves oblan- 7040 (holotype: JEPS!). ceolate to elliptic, subentire, sinuate-dentate, or, rarely, pinnatifid, 1–4(–5) cm long. Cauline leaves narrowly elliptic, widest Thysanocarpus trichocarpus Rydb., Bull. Torrey Bot. Club 30: near the middle, base cuneate, with small, inconspicuous 253. 1903.—TYPE: U. S. A. Utah: Washington: Silver Reef, auricles that do not extend around the stem, or, rarely, with 4 May 1894, E. L. Greene 5163b (holotypes: US!, isotype: conspicuous auricles extending around the stem. Flowers NY!, UC!). with yellow anthers. Lower fruiting pedicels 1.5–4(–5.5) mm, Trichomes absent. Fruits 3–5(–6) mm wide, the wings era- stiffly spreading or recurved in a continuous arc. Fruits usu- diate, entire or sparingly incised, the style 0.2–0.7 mm long. ally pulverulent with minute (0.05–0.1 mm) clavate hairs, Figure 9D . sometimes glabrous, flat or plano-convex, obovate to nearly Distribution— United States (northwestern Arizona, south- orbicular, 2.5–4 mm wide; wing shallowly to deeply crenate, ern California, northwestern Colorado, southern Nevada, rarely entire, usually with ± indistinct rays 0.2–0.5 mm wide. and southwestern Utah). Figure 10 . Figure 7B . Specimens intermediate between this subspecies and subsp. Distribution— United States, southern California (Inyo, amplectens occur in southern California and central Arizona, Kern, Los Angeles, Riverside, San Bernardino, and Santa and apparent hybrids with subsp. longistylus are found in Barbara Counties). Figure 8 . southern California (Kern and San Bernardino Counties). Munz named T. laciniatus var. hitchcockii based on the Subspecies eradiatus is, however, uniform outside these areas. minutely pubescent silicles of the type. Later, Munz (1959) Although T. trichocarpus is the older name and would have listed T. desertorum as a “probable synonym”, although the priority at species rank, this taxon is usually referred to in the silicles of type material of T. desertorum are glabrous. Among literature as var. eradiatus , so the latter name is used here at the specimens currently included in T. desertorum, most have subspecies level. The types of var. eradiatus and T. trichocarpus pubescent silicles but a minority have glabrous silicles; varia- differ in the pubescence of the silicle (glabrous in the former, tion in this character is unsurprising given that no taxon in the pubescent in the latter) but are otherwise similar. genus has uniformly pubescent silicles (see also discussion under T. curvipes). Although anther color has not been used 2 e. Thysanocarpus curvipes Hook. subsp. longistylus ( Jepson) previously in Thysanocarpus taxonomy, the yellow anthers P. J. Alexander & Windham, comb. nov. Thysanocarpus of T. desertorum provide a means of distinguishing glabrous- curvipes Hook. var. longistylus Jepson, Man. Fl. Pl. Calif. fruited plants from otherwise similar taxa, notably T. lacinia- 447. 1925.—TYPE: U. S. A. California: Tulare: foothills, tus and T. rigidus . upper San Joaquin, May 1907, W. L. Jepson 13383 (holotype: JEPS!). 4. Thysanocarpus erectus S. Watson, Proc. Amer. Acad. Arts Trichomes present or, rarely, absent. Fruits 3–4.5(–5) mm 11: 124. 1876.—TYPE: MEXICO. Baja California: Mcpio. wide, the wings indistinctly radiate and entire to distinctly Ensenada: Guadalupe Island, 1875, E. Palmer 9 (holotype: radiate and divided into spatulate lobes, or perforate, the GH, isotype: NY!). style 0.9–1.5(–1.8) mm long. Figure 9E . Trichomes absent or, rarely, plants sparsely hirsute at base. Distribution— United States, California (Tuolumne Co. Basal leaves oblanceolate to obovate, sinuate-dentate, 2–5(–6) south to Kern Co.). Figure 10 . cm long. Cauline leaves lanceolate, widest at base, auriculate- Although this subspecies was named based on style length clasping, often toothed, sometimes entire. Flowers with yel- alone, it overlaps slightly with subsp. elegans in this character. low anthers. Lower fruiting pedicels 2.5–5 mm long, erect to Nonetheless, subsp. longistylus is readily separated from that ascending proximally but often reflexed distally. Fruits with subspecies by its smaller, imperforate fruits. Subspecies longi- clavate hairs 0.2–0.4 mm long or glabrous, flat or plano-con- stylus occurs almost entirely within the range of subsp. cur- vex, nearly orbicular, (3–)4–5.5 mm wide; deeply notched at vipes , as mentioned above, but plants intermediate between base and apex, otherwise entire or occasionally shallowly the two are rarely collected. However, narrow-winged, rela- incised. Figure 7C . tively long-styled plants of subsp. eradiatus occurring along Distribution—Mexico, Baja California (Mcpio. Ensenada). the southern periphery of the distribution of subsp. longi- Figure 8 . stylus may result from introgression between these two sub- Although named for the erect pedicels of the type, the spe- species. SubspeciesProof longistylus is the only subspecies for ciesOnly is variable in this regard. Pedicels are usually erect or 16 SYSTEMATIC BOTANY [Volume 35 strongly ascending at the base, but may be strongly recurved ±0.2 mm long, or glabrous, flat, orbicular, 7–10 mm wide; distally. The fruits are much like those of T. curvipes subsp. wing entire or with undulate margins, with dark, narrow eradiatus, except that they are deeply notched at both the base (±0.1 mm) rays, these extending most or all of the way to the and apex, and the wings are often purplish. margins. Figure 7E . 5. Thysanocarpus laciniatus Nutt. ex Torr. & A. Gray, Fl. Distribution—United States, California and southern N. Amer. 1: 118.Proof 1840.—TYPE: U. S. A. California: Santa Oregon.Only Figure 8 — Barbara Co.: Santa Barbara, s. d., T. Nuttall s. n. (holotype: Perhaps the most visually distinctive taxon in the genus BM!, isotypes: NY!, PH!). due to the large, entire-winged fruits with distinct, narrow rays, this species occurs primarily in seasonally moist, low- Thysanocarpus emarginatus Greene, Pittonia 3: 86–87. 1896. elevation sites. Thysanocarpus radians often occurs with or Thysanocarpus laciniatus Nutt. ex Torr. & A. Gray var. near T. curvipes subsp. elegans, and occasional plants approach emarginatus (Greene) Jepson, Man. Fl. Pl. Calif. 447. 1925. T. curvipes subsp. elegans morphologically; whether this can Thysanocarpus curvipes Nutt. ex Torr. & A. Gray var. emar- be attributed to hybridization is uncertain. ginatus (Greene) Jepson, Fl. Calif. 2: 100. 9136.—TYPE: U. S. A. California: Contra Costa Co.: Mount Diablo, 7. Thysanocarpus rigidus (Munz) P. J. Alexander & Windham, 20 Jun 1892, E. L. Greene s. n. (holotype: NDG!). comb. nov. Thysanocarpus laciniatus Nutt. ex Torr. & A. Gray var. rigidus Munz, Bull. S. Calif. Acad. Sci. 31: 62. Trichomes absent or, rarely, plants sparsely hirsute at base. 1932.—TYPE : U. S. A. California: San Diego Co.: Laguna Foliage usually greenish, sometimes purplish at base. Basal Camp, Laguna Mountains, 16 May 1925, P. A. Munz 9701 leaves oblanceolate to elliptic, often pinnatifid, sometimes sin- (holotype: POM!). uate-dentate or subentire, 2–6(–9) cm long. Cauline leaves linear to narrowly elliptic, widest near the middle or equally wide Trichomes absent. Foliage purplish throughout. Basal throughout, base cuneate or with small, inconspicuous auri- leaves oblanceolate to elliptic, subentire, sinuate-dentate, or cles that do not extend around the stem. Flowers with purple pinnatifid, 2–5 cm long. Cauline leaves linear to narrowly anthers, or rarely some flowers with infertile, whitish anthers. elliptic, widest near the middle or equally wide throughout, Lower fruiting pedicels 2.5–6(–8) mm long, recurved in a con- base cuneate or with small, inconspicuous auricles that do not tinuous arc. Fruits usually glabrous, sometimes with clavate extend around the stem. Flowers with purple anthers. Lower hairs 0.05–0.4 mm long, flat or plano-convex, obovate to nearly fruiting pedicels 1.5–6 mm long, straight and stiffly spread- orbicular, 3–5 mm wide; wing entire to deeply crenate, eradi- ing to ascending. Fruits glabrous, obovate to nearly orbicular, ate or with ± indistinct rays 0.2–0.5 mm wide. Figure 7D. 2.5–3.5 mm wide; wing entire to shallowly crenate, eradiate or Distribution—United States (southern California, west- with ± indistinct rays 0.2–0.5 mm wide. Figure 7F . ern Nye County in Nevada) and Mexico (northern Baja Distribution— United States, southern California (Riverside, California). Figure 8 . San Bernardino, and San Diego Counties) and Mexico (north- As discussed above, specimens assigned to T. laciniatus ern Baja California). Figure 8 . are allopolyploids combining genomes from the conchulif- Thysanocarpus rigidus is poorly represented in herbarium erus clade and the curvipes clade. Thysanocarpus laciniatus also collections, with only five specimens known. This species has had multipe independent hybrid origins which are not can be difficult to distinguish from T. laciniatus, which occa- given separate taxonomic recognition here pending further sionally has the branched, purplish appearance of T. rigidus . investigation. The stiff, straight, horizontal to slightly ascending pedicels of Distinguishing T. laciniatus and T. curvipes subsp. amplect- T. rigidus allow identification in these cases. This species ens can be difficult in the vicinity of mainland Los Angeles occurs in arid mountain ranges up to moderately high ele- County and the Channel Islands in California. The presence vations (2,150 m on New York Peak), and three of the five of auriculae on the cauline leaves appears to be a nearly con- specimens were collected in May, when little else is likely to tinuous rather than discrete character in plants from this area. be flowering. Poor sampling of these habitats at this time of Unfortunately, no other reliable morphological characters year may account for the paucity of specimens, but T. rigidus that distinguish the two taxa are known. It is likely that there may be of conservation concern and should be investigated is ongoing hybridization between the two taxa in this area, further. Additional Specimens— MEXICO. Baja California: Mcpio. Ensenada: and some specimens cannot confidently be assigned to either 50 miles SE of Tecate, 13 May 1925, P. A. Munz 9572 (POM!). T. laciniatus or T. curvipes subsp. amplectens . U. S. A. California: Los Angeles Co.: North of Claremont, 14 Mar 1923, 6. Thysanocarpus radians Benth., Pl. Hartw. 297. 1849.— P. A. Munz 6589 (POM!). California: Riverside Co.: SSW of Palm Desert along CA 74 near rim of Deep Canyon ca. 0.57 km SSW of the summit TYPE: U. S. A. California: Sacramento Co.: Sacramento, (4775) of Sugarloaf Mountain, 27 Feb 2005, M. D. Windham 3299 (DUKE!, s. d., Hartweg 1651 (holotype: K!, isotypes: GH, NY!). MO!, NMC!, UT!). California: San Bernardino Co.: New York Mts.; E slope Thysanocarpus radians Benth. var. montanus Jepson, Fl. W. Calif. of New York Peak, R. F. Thorne 44490 (UC!). 226. 1901.—TYPE : U. S. A. California: Napa: Plateau of Acknowledgments. We would like to thank the following institu- the Napa Mountains, north of Mt. George, 28 Apr 1893, tions for loans of specimens: California Academy of Sciences (CAS, DS), W. L. Jepson 30d (holotype: JEPS!). New York Botanical Garden (NY), Rancho Santa Ana Botanic Garden (POM, RSA), University of California, Berkeley (JEPS, UC), University Trichomes absent, or plants rarely sparsely hirsute at of Notre Dame (NDG), and Utah Museum of Natural History (UT). We base. Basal leaves oblanceolate, sinuate dentate to runcinate- would also like to thank the following institutions for label information pinnatifid, 1.5–4 cm long, usually glabrous. Cauline leaves and/or photographs of type specimens: The Academy of Natural Sciences, lance-ovate to lanceolate, widest at base, auriculate-clasping. Philadelphia (PH), Botanical Garden of the V. L. Komarov Institute of the Flowers with yellow anthers. Lower fruiting pedicels 7–18 Russian Academy of Sciences (LE), and Natural History Museum, London (BM). Thanks also to Dr. K. Gandhi for nomenclatural assistance, and to mm, weakly ascending, straight or nearly so but with genic- Dr. N. H. Holmgren and one anonymous reviewer for helpful comments. ulately reflexed tips.Proof Fruits usually with clavate trichomes ThisOnly research was supported by NSF DEB-0817033 & EF-0542228 (CDB). 2010] ALEXANDER ET AL.: THYSANOCARPUS PHYLOGENETICS AND TAXONOMY 17

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All Thysanocarpus sequences were generated for the Hooker, W. J. 1836 . Icones Plantarum . London : Longman, Rees, Orme, present study. Among other taxa, accessions from which sequences were Brown, Green, and Longman . generated for the present study are marked with an asterisk. Hughes, C. E., C. D. Bailey , and S. A. Harris . 2002 . Divergent and reticulate species relationships in Leucaena (Fabaceae) inferred from multiple Thelypodieae: Thysanocarpus— 1. Thysanocarpus conchuliferus, U. S. A.: data sources: insights into polyploid origins and nrDNA polymor- California: Santa Barbara Co.: Santa Cruz Island, rocky butte E of Lagunita phism . American Journal of Botany 89: 1057 – 1073 . Secas (originally M.R. Benedict, 28 May 1969; grown under lath at Santa McNeill, J., F. R. Barrie , H. M. Burdet , V. Demoulin , D. L. Hawksworth , Barbara Botanic Garden), R.B. Philbrick B69-130 (DS), GU246121, GU246184. K. Marhold , D. H. Nicolson , J. Prado , P. C. Silva , J. E. Skog , 2. Thysanocarpus conchuliferus, U. S. A.: California: Santa Barbara Co.: Santa J. H. Wiersema , and N. J. Turland . 2006 . International Code of Cruz Island, Twin Harbor, R. Hoffman 655 (RSA), GU246122, GU246185. Botanical Nomenclature (Vienna Code), adopted by the Seventeenth 3. Thysanocarpus conchuliferus, U. S. A.: California: Santa Barbara Co.: Santa International Botanical Congress Vienna, Austria, July 2005. Regnum Cruz Island, upper west fork of Orizaba Canyon (fork which intersects Vegetabile 146 . Ruggell, Lichtenstein : A. R. G. Gantner Verlag K. G . main ridge line due S of Frye’s Harbor), S. A. Junak 1053 (RSA), GU246123, Müller, K. 2005 . SeqState: primer design and sequence statistics for phylo- GU246186. 4. Thysanocarpus curvipes subsp. amplectens , Mexico: Sonora: genetic DNA datasets . Applied Bioinformatics 4: 65 – 69 . Mcpio. Tubutama: 4.5 km SE of Tubutama on road to Magdalena, T. R. Van Mummenhoff, K., I. A. Al-Shehbaz , F. T. Bakker , H. P. Linder , and Devender 2003-262 (NMC), GU246124, GU246187. 5 . Thysanocarpus curvipes A. Mülhausen . 2005 . Phylogeny, morphological evolution, and spe- subsp. amplectens , U. S. A.: Arizona: Cochise Co.: South-southwest of ciation of endemic Brassicaceae genera in the Cape flora of Southern Willcox along Stronghold Canyon East in the Dragoon Mountains ca. Africa . Annals of theProof Missouri Botanical Garden 92: 400 – 424 . 1.93Only km southeast of Barret Spring, M. D. Windham 2929 (NMC), GU246125, 18 SYSTEMATIC BOTANY [Volume 35

GU246188. 6. Thysanocarpus curvipes subsp. amplectens , U. S. A.: Arizona: Pinyon Hills, Highway 138 junction with Highway 18, S.Boyd 1540 (UC), Graham: Camp at confluence of Bonita Creek and Gila River, L.A. McGill ITS only: GU246151. 33. Thysanocarpus desertorum, U. S. A.: California: Los 1968 (CAS), GU246126, GU246189. 7 . Thysanocarpus curvipes subsp. Angeles Co.: Big Rock Creek ca. 0.25 miles south of Big Pines Highway, amplectens , U. S. A.: Arizona: Greenlee Co.: East of Clifton near Arizona between the stream and Big Rock Creek Road; Valyermo, T. S. Ross 6100 Highway 78 on slope overlooking Black Jack Canyon ca. 1.29 km north- (CAS), GU246152, GU246215. 34. Thysanocarpus desertorum , U. S. A.: east of Dunigan Spring, M. D. Windham 2921 (NMC), GU246127, GU246190. California: Riverside Co.: Joshua Tree National Monument, Squaw Tank, 8. Thysanocarpus curvipes, intermediate between subsps. amplectens and J. H. Thomas 18477 (DS), GU246153, GU246216. 35. Thysanocarpus erectus , eradiatus , U. S. A.: Arizona:Proof La Paz Co.: Highway 72 17.5 km east of Salome, Mexico:Only Baja California: Mcpio. Ensenada: 55 miles southeast of San Luis 12.5 km W of Maricopa Co. line, R. W. Spellenberg 13138 (NMC), GU246128, Gonzaga, J. Henrickson 19339 (RSA), GU246154, GU246217. 36. Thys- GU246191. 9. Thysanocarpus curvipes, intermediate between subsps. anocarpus erectus, Mexico: Baja California: Mcpio. Ensenada: 22 miles amplectens and eradiatus, U. S. A.: Arizona: La Paz Co.: East of Wenden on northeast of El Arco, on the road to San Francisquito, S. Boyd 5756 (RSA), lower slopes of the Harquahala Mountains ca. 2.81 km northwest of the GU246155, GU246218. 37. Thysanocarpus erectus, Mexico: Baja California: summit (5,681) of Harquahala Peak, M. D. Windham 3290A (NMC), Mcpio. Ensenada: East of La Ramona, S. Boyd 5381 (RSA), GU246156, GU246129, GU246192. 10 . Thysanocarpus curvipes subsp. eradiatus , U. S. A.: GU246219. 38. Thysanocarpus erectus , Mexico: Baja California: Mcpio. Arizona: Mohave Co.: South side of Virgin River Gorge on slopes above Ensenada: North of Loma Creston Prieto, S. Boyd 5441 (RSA), trnL-F only: Interstate 15 ca. 1.0 road miles east of the lower entrance to The Narrows, GU246220. 39. Thysanocarpus laciniatus , U. S. A.: California: Alameda Co.: M. D. Windham 97-052 (NMC), GU246130, GU246193. 11. Thysanocarpus Southeast of Livermore, along Mines Road (Arroyo Mocho) ca. 13.5 miles curvipes subsp. amplectens , U. S. A.: Arizona: Pima Co.: Rincon Mountains, south of the junction with Tesla Road, L. R. Heckard 2507 (JEPS), GU246157, Saguaro National Monument East, Tanque Verde Ridge Trail, C. E. Jenkins GU246221. 40. Thysanocarpus laciniatus, U. S. A.: California: Kern Co.: 1272 (MO),GU246131, GU246194. 12. Thysanocarpus curvipes subsp. Flying Dutchman Creek canyon on Havilah Mountain Breckenridge road, amplectens, U. S. A.: Arizona: Pima Co.: Lower Thomas Canyon, 4 miles J. T. Howell 47794 (CAS), GU246158, GU246222. 41. Thysanocarpus lacinia- southeast of Baboquivari Peak in the Baboquivari Mountains, P. J. tus , U. S. A.: California: Los Angeles Co.: North of Azusa along West Alexander 93 (NMC), GU246132, GU246195. 13. Thysanocarpus curvipes Fork of the San Gabriel River ca. 1.51 km north of Fern Springs, subsp. amplectens , U. S. A.: Arizona: Pima Co.: Coronado National Forest M. D. Windham 98-103 (NMC), 41A: GU246160, 41B: GU246159; GU246223. ca. 9.0 miles north of Tucson on the Catalina Highway, H. H. Schmidt 2562 42. Thysanocarpus laciniatus , U. S. A.: California: Los Angeles Co.: San (MO), GU246133, GU246196. 14 . Thysanocarpus curvipes subsp. amplectens , Francisquito Canyon, westerly draw draining from ridge which divides S. U. S. A.: Arizona: Santa Cruz Co.: North side of the Pajarito Mountains on Fr. Cyn. from Dry Canyon; ca. 630 m south of where Drinkwater the west side of Peña Blanca Lake, P. J. Alexander 89 (NMC), GU246134, Canyon joins S. Fr. Cyn., T. S. Ross 7472 (CAS), GU246161, GU246224. GU246197. 15. Thysanocarpus curvipes subsp. curvipes , U. S. A.: California: 43. Thysanocarpus laciniatus , U. S. A.: California: Los Angeles Co.: San Calaveras Co.: Highway 49 1.6 km north of the junction with Hwy. 120, Clemente Island, east side of the island near Malo Point, first canyon south north of Chinese Camp, south side of North Fork of the Calaveras River, of Bryce Canyon, S. Boyd 4301 (CAS), GU246162, GU246225. R. W. Spellenberg 13504 (NMC), GU246135, GU246198. 16. Thysanocarpus 44. Thysanocarpus laciniatus , U. S. A.: California: Orange Co.: Along tribu- curvipes subsp. curvipes , U. S. A.: California: Colusa Co.: Goat Mountain tary of Hot Spring Canyon in the Santa Ana Mountains ca. 2.91 km south- Road at Sullivan Creek, 1.2 miles southwest of Digger Pine Flat, south east of Crow Spring, M. D. Windham 98-095 (MO), 44A: GU246163, 44B: of Stonyford, V. H. Oswald 6064 (JEPS), GU246136, GU246199. 17. GU246164; GU246226. 45. Thysanocarpus laciniatus, U. S. A.: California: San Thysanocarpus curvipes subsp. eradiatus, U. S. A.: California: Imperial Co.: Bernardino Co.: North-northwest of Yucca Valley in Water Canyon along Southwest of Ocotillo in the Jacumba Mountains near Interstate 8 ca. Pioneertown Rd. ca. 1.5 road miles north-northwest of its junction 1.35 km east-northeast of Mountain Spring, M. D. Windham 3295 (NMC), with CA Highway 62, M. D. Windham 2957A (NMC), GU246165, GU246227. GU246137, GU246200. 18. Thysanocarpus curvipes subsp. elegans , U. S. A.: 46. Thysanocarpus laciniatus , U. S. A.: California: San Diego Co.: West of California: Mendocino Co.: 8 km south of Yorkville, G. L. Smith 6490 (CAS), Campo along old CA Highway 94 on slope above Campo Creek ca. 2.03 GU246138, GU246201. 19. Thysanocarpus curvipes subsp. elegans , U. S. A.: km north of VABM 3053 on the US-Mexico border, M. D. Windham 3424 California: San Diego Co.: Southeast of Julian along Sunrise Highway near (NMC), 46A: GU246166, 46B: GU246167; GU246228. 47. Thysanocarpus lac- Lake Cuyamaca ca. 2.80 miles east-southeast of the summit (5,593) of iniatus , U. S. A.: California: Santa Barbara Co.: Santa Cruz Island, mouth of North Peak, M. D. Windham 2948 (NMC), GU246139, GU246202. 20. Dick’s Harbor, S. A. Junak SC-1473 (UC), 47A: GU246169, 47B: GU246168; Thysanocarpus curvipes subsp. elegans, U. S. A.: California: Sutter Co.: South GU246229. 48. Thysanocarpus laciniatus , U. S. A.: California: Sutter Co.: side of South Butte, Sutter Buttes, L. Ahart 2726 (CAS), trnL-F only: South Butte, Sutter Buttes; along the edge of the road to the lower radio GU246140, GU246203. 21. Thysanocarpus curvipes subsp. elegans , U. S. A.: tower on the South Butte, L. Ahart 4637 (CAS), 48A: GU246170, 48B: California: Tehama Co.: BLM parcel along Highway 36, east edge of GU246171, GU246230. 49. Thysanocarpus laciniatus, U. S. A.: Nevada: Nye Paynes Creek near its mouth on the Sacramento River, Hog Lake Plateau, Co.: Northwest of Rhyolite, M. J. Williams 78-37-1 (CAS), 49A: GU246172, V. H. Oswald 7332.1 (JEPS), GU246141, GU246204. 22. Thysanocarpus curvi- 49B: GU246173, GU246231. 50. Thysanocarpus radians, U. S. A.: California: pes subsp. longistylus, U. S. A.: California: Tuolumne Co.: Along north Butte Co.: Circa 2 air miles north-northeast of Honcut between Oroville bank of North Fork of Tuolumne River, 2.9 miles from Long Barn, I. L. and Marysville, Peter Ahart Ranch, B. Ertter 5884 (UC), GU246174, Wiggins 21272 (DS), GU246142, GU246205. 23. Thysanocarpus curvipes GU246232. 51. Thysanocarpus radians , U. S. A.: California: Napa Co.: subsp. longistylus , U. S. A.: California: Tuolumne Co.: Carnegie Institute of 6 miles north of Monticello, C. Best 914 (CAS), GU246175, GU246233. Washington Field Station at Mather, F. Almeda 4365 (CAS), GU246143, 52. Thysanocarpus radians , U. S. A.: California: Sacramento Co.: Intersection GU246206. 24. Thysanocarpus curvipes subsp. curvipes , U. S. A.: Idaho: Ada of Bradshaw and Shelton Roads, ca. 3 miles northeast of Elk Grove, G. H. Co.: Freestone Creek, northeast of Boise, B. Ertter 3231 (CAS), GU246144, True 6516 (CAS), GU246176, GU246234. 53. Thysanocarpus radians , U. S. A.: GU246207. 25. Thysanocarpus curvipes subsp. amplectens , U. S. A.: New California: Tehama Co.: Northwest side of Sacramento Valley, ca. Mexico: Luna Co.: A small canyon on the east side of the narrow central 0.75 miles south of Gyle Road, V. H. Oswald 7240 (MO), GU246177, portion of the Florida Mountains, just south of Lobo Draw, P. J. Alexander GU246235. 54. Thysanocarpus rigidus , U. S. A.: California: Riverside Co.: 188 (NMC), GU246145, GU246208. 26. Thysanocarpus curvipes subsp. era- SSW of Palm Desert along CA 74 near rim of Deep Canyon ca..57 km SSW diatus, U. S. A.: Nevada: Clark Co.: West of Las Vegas along Pine Creek of the summit (4775) of Sugarloaf Mountain, M. D. Windham 3299 (NMC), Canyon in the Spring Mountains ca. 0.31 km north-northeast of Pine Creek GU246178, GU246236. 55. Thysanocarpus rigidus , U. S. A.: California: San Spring, M. D. Windham 98-118 (NMC), trnL-F only: GU246209. 27. Thys- Bernardino Co.: New York Mountains; east slope of New York Peak, anocarpus curvipes subsp. curvipes , U. S. A.: Nevada: Humboldt Co.: Calico R. F. Thorne 44490 (UC), GU246179, GU246237. Mountains, A. Tiehm 13112 (CAS), GU246146, GU246210. 28. Thysanocarpus Other Thelypodieae— 56. Caulanthus inflatus S. Watson, AF531617, curvipes subsp. curvipes, U. S. A.: Nevada: Washoe: Virginia Mountains, AY958534. 57. Caulanthus lasiophyllus Payson, AF346644, AY958535. Paiute Canyon at south end of the range, 0.4 road miles north of main foot- 58. Chilocardamum castellanosii (O. E. Schulz) Al-Shehbaz, AY958592, hill road, A. Tiehm 13778 (CAS), GU246147, GU246211. 29. Thysanocarpus AY958548. 59. Coelophragmus auriculatus O. E. Schulz, AF531598, AY958547. curvipes subsp. curvipes, U. S. A.: Nevada: Washoe: Steamboat Hot Springs, 60. Dryopetalon runcinatum A. Gray, AF531634, AY958536. 61. Hesperidanthus J. T. Howell 48663 (CAS), GU246148, GU246212. 30. Thysanocarpus curvipes argillaceus (S. L. Welsh & N. D. Atwood) Al-Shehbaz, AF531611, AY958542. subsp. eradiatus , U. S. A.: Utah: Washington Co.: Southeast slope of the 62. Hesperidanthus linearifolius (A. Gray) Rydb., AF531612, AY958540. Pine Valley Mountains above Leeds Creek ca. 2.78 km south of Italian 63. Mostacillastrum andinum (Phil.) Al-Shehbaz, AF531650, AY958546. Spring, M. D. Windham 97-056 (NMC), GU246149, GU246213. 31. Thys- 64. Mostacillastrum commune (Speg.) Al-Shehbaz, AF531599, AY958564. anocarpus curvipes subsp. curvipes, U. S. A.: Washington: Klickitat Co.: Near 65. Mostacillastrum gracile (Wedd.) Al-Shehbaz, AY958596, AY958554. Klickitat Co. landfill, R. R. Halse 3394 (UC), GU246150, GU246214. 66. Mostacillastrum leptocarpum (Hook. & Arn.) Al-Shehbaz, AF531632, 32. Thysanocarpus desertorumProof , U. S. A.: California: Los Angeles Co.: East of AY958556.Only 67. Mostacillastrum subscandens (Speg.) Al-Shehbaz, AF531631, 2010] ALEXANDER ET AL.: THYSANOCARPUS PHYLOGENETICS AND TAXONOMY 19

AY958567. 68. Neuontobotrys frutescens (Gillies ex Hook. & Arn.) Al-Shehbaz, AJ628339+AJ628340, DQ180301. 91.* Athysanus pusillus, U. S. A.: Arizona: AY958595, AY958551. 69. Polypsecadium effusum (O. E. Schulz) Al-Shehbaz, Gila Co.: Southwest of Globe along road to Pinal Recreation Area in the AY958594, AY958550. 70. Polypsecadium magellanicum (Pers.) Al-Shehbaz, Pinal Mountains, ca. 4.21 km north-northwest of the summit (7812) of AF531589, AY958558. 71. Romanschulzia arabiformis (DC.) Rollins, AF531635, Signal Peak, M. D. Windham 2937 (NMC), GU246183, GU246241. 92. Draba AY958538. 72. Romanschulzia costaricensis (Standl.) Rollins, AF531636, nemorosa L., AY047676, DQ467041. 93. Boechera stricta (Graham) Al-Shehbaz, AY958539. 73. Sibara laxa Greene, AF531637, AY958543. 74. Stanleya pin- AF183114, AF307554. 94. Brassica juncea (L.) Czern., DQ340651, EF426774. nata (Pursh) Britton, AF531620, AY122459. 75.* Streptanthella longirostris , 95. Crucihimalaya wallichii (Hook. f. & Thomson) Al-Shehbaz, O’Kane, & U. S. A.: Utah: WayneProof Co.: West of Teasdale at N base of Boulder Mtn. R. OnlyA. Price, AF137555, DQ310520. 96. Barbarea vulgaris W. T. Aiton, ca. 4.04 km SSW of the summit (7472) of Velvet Ridge, M. D. Windham AJ232915, AY122453. 97. Cochlearia pyrenaica DC., DQ780946+DQ780947, 97-058 (MO), GU246180, GU246238. 76. Streptanthus campestris S. Watson, DQ180269. 98.* Descurainia californica O. E. Schulz, U. S. A.: Utah: Beaver AF346646, AY958571. 77. Streptanthus cordatus Nutt. ex Torr. & A. Gray, Co.: Circa 13 miles east-northeast of Milford, Bailey’s Spring, S. L. Welsh AF531622, AY958572. 78. Streptanthus glandulosus Hook., AF346652, 13949 (MO), GU246181, GU246239. 99. Neotorularia korolkowii (Regel & AY958575. 79. Thelypodiopsis elegans Rydb., AF531638, AY958578. Schmalh.) Hedge & J. Léonard, AY353156, AY353230. 100. Sphaerocardamum 80. Thelypodiopsis shinnersii (M. C. Johnst.) Rollins, AF531640, AY958579. nesliiforme S. Schauer, AF055195, AF307529. 101. Heliophila coronopifolia L., 81. Thelypodiopsis vaseyi (S. Watson ex B. L. Rob.) Rollins, AF531641, DQ249846, DQ518369. 102. Hesperis matronalis L., AJ628313+AJ628314, AY958580. 82. Thelypodium flexuosum B. L. Rob., AF531624, AY958582. AY546166. 103. Iberis amara L., AJ440311, AY122455. 104. Isatis tinctoria L., 83. Thelypodium wrightii A. Gray, AF531615, AY958583. 84. Warea AF384104, DQ518370. 105. Lepidium virginicum L., AY662280, DQ821405. amplexifolia Nutt., AF531642, AY958584. 85. Warea cuneifolia Nutt., AF531643, 106. Noccaea fendleri (A. Gray) Holub, AY154819, AY154804. 107.* Lyrocarpa AY958585. 86. Weberbauera peruviana (DC.) Al-Shehbaz, AF531593, coulteri Hook. & Harv., U. S. A.: Arizona: Pima Co.: East of State Route 85 AY958562. 87. Werdermannia anethifolia I. M. Johnst., AF531645, AY958586. near the base of the Gunsight Hills ca. 7.53 km east-northeast of Armenta Other tribes, alphabetically by tribe— 88. Aethionema grandiflorum Well, M. D. Windham 98-038 (MO), GU246182, GU246240. 108. Sisymbrium Boiss. & Hohen., DQ249867, AP009367. 89. Lobularia libyca Webb & irio L., AF531558, AY958555. 109. Smelowskia calycina (Stephan ex Willd.) Berthel., DQ249847, DQ518372. 90. Matthiola incana (L.) W. T. Aiton, C. A. Mey., AY230640, DQ180295.

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