Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Taxonomists. All rights reserved. O 10.1600/036364413X670313 DOI © Botany Systematic ayeooial motn lns uha rosemary as such , important of ( contains and consists economically 2012) Sytsma and many Tribe Drew to Sytsma 2012). (according and genera Sytsma 67 (Walker and Drew the mesas. 2006; of on Menthinae found subtribe typically pool are vernal as which in known below), occurrence commonly (see common are their habitats from the mints, Lamiaceae of mesa family Members plant 1). the (Table of species recognized currently .T Howell, T. J. Pogogyne Hedeomoides aite aebe ecie nteps,alo hc etreat we which or of subspecies all past, several the species and in endangered described species been federally have additional as varieties two Two listed and 1). also species, are (Table endangered three these as of listed are these of nudiuscula P. pclpoes ay ettr,crlalnt n relative number 2012). and al. fertile et and length (Silveira calyces, corolla and vestiture, from exsertion and calyx shape lobe length process, calyx length, (including apical lobe stem calyx shape and including calyx width), and features, habit, morphological and diameter several Mexico Idaho; California, on of possibly Baja based species northern (and Different in 2). Oregon and (Fig. 2006), in Meinke see populations see however, 1993; some (Jokerst, the with fruit on the trichomes of Species on with Discussion). and annuals 1E) being (Fig. in style tribe the other in from distinguished genera are but 1A–E) (Fig. flowers bilabiate Pogogyne, cataria eaiet h nlrsec rcs(agradexserted and (larger bracts inflorescence subgen. the in to relative subgen. subgen. in in (four two versus fertile of number the in differ subgenus subgenus omrnsofficinalis Rosmarinus oyih 03b h mrcnSceyo ln Taxonomists Plant of Society American the by 2013 Copyright Pogogyne Pogogyne eainhp fti ru,DAsqec aafo the from data sequence DNA group, this of relationships ut hr rnhs u siaefrtema iegnetm ftese oei .–. ilo er g n o hto h rw node crown the members of the that that hypothesis for the and support ago results years These million ecosystems. 5.1–7.7 pool vernal is of node ages stem with the overlapping latter of of the time divergence ago, years mean million the 0.9–1.9 for is estimate Our branches. Subgenus short size. withinquite corolla groups methods. reduced separate reconstruction a phylogeny to Pogogyne Bayesian and used and number been parsimony stamen have both subgenus that in using characters that morphological analyzed indicates for and performed study outgroups were several reconstructions and state species Ancestral extant seven all from Abstract— Keywords— Pogogyne .,adsg ( sage and L.), scasfe nsbaiyNe subfamily in classified is a orspecies: four has iems ebr ftefml,hv zygomorphic, have family, the of members most like Hedeomoides Pogogyne e oes,19;Sler ta.21) Subgenus 2012). al. et Silveira 1993; Jokerst, see ; 21) 83:p.782–794 pp. 38(3): (2013), Pogogyne et.i ml eu fegt(ee extant) (seven eight of genus small a is Benth. sotndvddit w nrgnrcgroups, infrageneric two into divided often is aedsrt igorpi itiuin.The distributions. biogeographic discrete have .Ga.(e i.2– o itiuin. Three distributions.) for 2A–C Fig. (See Gray. A. .douglasii P. newn ai iesfcto nrsos oseilzto oaproial nnae habitat. inundated periodically a to specialization to response in diversification rapid a underwent Pogogyne , hlgntcSseaiso h eaMints: Mesa the of Systematics Phylogenetic eateto ilg,SnDeoSaeUiest,SnDeo aiona912 .S A. S. U. 92182, California Diego, San University, State Diego San Biology, of Department [subgen. .,mn ( mint L.), esssaladicue nsubgen. in included and small versus Hedeomoides Salvia .Ga Tbe1.Tetosubgenera two The 1). (Table Gray A. Lmaee sasalgnso nulpat,ocrigams nieyi enlpo aias oifrtephylogenetic the infer To habitats. pool vernal in entirely almost occurring plants, annual of genus small a is (Lamiaceae) Pogogyne et.(h yefrtegns,and genus), the for type (the Benth. .abramsii P. pce) ebr ftegenus the of Members species). Pogogyne iegne etee Menthinae, Mentheae, divergence, Eupogogyne Mentha Hedeomoides n xeto ftecorolla the of exsertion and ) cu otyi California, in mostly occur 1 eoda,tieMentheae, tribe petoideae, uhrfrcrepnec ([email protected]) correspondence for Author ihe .Sler n ihe .Simpson G. Michael and Silveira A. Michael .T Howell, T. J. pce) anp( catnip species), aebe diagnosed been have smnpyei ihgo upr n a edansdb w pmrhe:areduction a apomorphies: two by diagnosed be can and support good with monophyletic is .T oel,and Howell], T. J. omnctn dtr nraWeeks Andrea Editor: Communicating .clareana P. Pogogyne Pogogyne trnQ-rps16 Pogogyne Nepeta eu ld a eaieyln tmlnae u t rw ld has clade crown its but lineage, stem long relatively a has clade genus sprpyei,btwt orspot eea ldswti h genus the within clades Several support. poor with but paraphyletic, is 782 hools pcradbt T n T ula ein eeobtained were regions nuclear ITS and ETS both and spacer chloroplast ta.20) ntehsoia at enlposwr much were pools vernal Meester past, (De historical period the period dry In a long 2005). a survive al. by et to followed adaptations inundation, of specific possess wet habitats seasonally generally would as which all encompass such streams, intermittent environments, and meadows similar term, vernal and broader both A include pools to 2003). used sometimes (Zedler is coast wetland, region ephemeral pool west climate American vernal Mediterranean the a term within on the those include the restrict only during definitions to precipitation Some “clay of season. pan,” retention wet allowing surface, and “hard ground water- accumulation the a a below for just have (e.g. flow) a basalt typically substrate or and pools pan,” of Vernal period layer year. inundated a shallow impermeable an a in as period both defined dry to generally is subjected pool” depression “vernal A 1F). (Fig. process the 2006). in (Meinke still investigated are see new being but of suggested, possible 2012; been have Two recognized al. California history). Baja et been subgenus taxonomic of Silveira not full species 1993; for has (Jokerst 2010 classification Silveira treatments this recent but Subgenus in extinct. 1), presumed sage- (Table is among species rare, the Hedeomoides and “very side.” since, as eastern found notes the (1875), collection on Watson brush, by the listed cited was who species This Mexico. California, .tenuiflora P. species: four ssnnm of synonyms as ( has there. species This 2E). (Fig. notbeenfoundinMexicosinceandispresumedextirpated Mexico California, his- Baja distributions). single, Quintin, a San for plus ( California 2D–F collection northern torical Fig. and central see in collected 2012; al. serpylloides Pogogyne et Silveira 1993; section as to referred and genus .Ple 65, Palmer E. enlpool. vernal , Most ooyetenuiflora Pogogyne Hedeomoides Pogogyne a entetda genus as treated been has .Ga,and Gray, A. 85 H0019)fo udlp sad Baja Island, Guadalupe from 00001496) GH 1875, Pogogyne .floribunda P. Pogogyne .douglasii P. rut1361 Orcutt pce r on nvra olhabitats pool vernal in found are species as spelled (also sdcmne rmnmru vouchers numerous from documented is Hedeomoides 1 aias lnsadaiaso these of animals and Plants habitats. skonsll rmoecollection one from solely known is Sler ta.21;Tbe1.Sub- 1). Table 2012; al. et (Silveira (Lamiaceae) .zizyphoroides P. Hediomoides Jokerst, ooyetenuiflora Pogogyne 1Ar18,U 59)from 25599) UC 1886, Apr 21 , Hediomoides rmOeo n northern and Oregon from .serpylloides P. nJkrt19)has 1992) Jokerst in Hedeomoides eta (Jokerst Bentham Pogogyne nHwl 1931 Howell in a o been not has .This .Gray, A. Briquet Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. Subgenus endangered; threat); of immediacy are: sources listing environmental Primary ohcae en elspotd rwadStm (2012) with Sytsma results, and similar Drew obtained well-supported. being clades both of analysis, a in (2006), Sytsma speciesof and one included Mentheae, Walker tribe the of recently. study phylogenetic done been have n naieseis(eMetre l 05 oln and Holland 2005; al. et Meester pollution 1995). a (De as Keil species such at invasive impacts, vanish for other and (particularly to and destruction development) habitat continuing direct human are to due lands pace Zedler and rapid unique (Keeley today These only exist but still 1998). to Province, thought Floristic are 10% California about the in common more Subgenus OF SYSTEMATICS PHYLOGENETIC SIMPSON: AND SILVEIRA 2013] elsewhere; listing; ihnte,adt ne n vlaeapcso character of aspects evaluate and infer to and them, two the of within of monophyly subgenera the test accepted to used are the trees and phylogenetic markers of DNA ribosomal species nuclear trnQ-rps16 extant ITS and seven ETS the using of relationships netic investigating studies genus the phylogenetic within relationships published previously villosa two these of consisting and Jokerst two of clade Mentheae, the of of species analysis different detailed a and more included Drew a 2006). polytomy in Sytsma (2011), a and Sytsma (Walker in genera embedded other was twelve of genera two these containing lanceolata this placed .zizyphoroides P. .tenuiflora P. .serpylloides P. floribunda P. .nudiuscula P. .clareana P. abramsii P. † ihlvlpyoeei tde htincluded that studies phylogenetic High-level Table h rmr betv fti td st ne h phyloge- the infer to is study this of objective primary The Pogogyne .douglasii P. cnhmnh lanceolata Acanthomintha [ [ [ [ [ [ [ [ [ [ [ [ [ Hedeomoides .s. P. Hedeomoides s. Hedeomoides serpylloides Hedeoma Hedeomoides .parviflora P. .d. P. d. P. .d. P. d. P. d. P. .multiflora P. sn hools N aaaoe hr r no are There alone. data DNA chloroplast using , Fed .Cretyrcgie ugnr n pce of species and subgenera recognized Currently 1. subsp. var. subsp. var. subsp. var. Hedeomoides Pogogyne ooyedouglasii Pogogyne 4 urn(otta upr au 7;teclade the 67); = value support (bootstrap Curran eea nagrdSeisAt(EA itn;seCP 02.Symbols: 2012). CNPS see listing; (FESA) Act Species Endangered Federal = FE pcrrgo fclrpatDA h resultant The DNA. chloroplast of region spacer iie itiuin(ac List); (Watch distribution Limited = .T oel orSaos4 2 1973 22. 4: Seasons Four Howell, T. J. .T oel rc ai.Aa.Si 0 1.1931 119. 20: Sci. Acad. Calif. Proc. Howell, T. J. multiflora tricolor parviflora .lanceolata A. ( .Ga,Po.Ae.Aa.At 1 0.17 ( 1876 100. 11: Arts Acad. Amer. Proc. Gray, A. .floribunda P. et. ait e.Se.44 1834 414. Spec. Gen. Labiat. Benth., Sade)Jkrt ls 32:37 1992 347. 13(2): Aliso Jokerst, (Standley) Pogogyne .Ga,Bt aiona[..rwr :57 1876 597. 1: [W.H.Brewer] California Bot. Gray, A. eeal endangered federally = intermedia minor ramosa et. ait e.Se.44 1834] 414. Spec. Gen. Labiat. Benth., et. ait e.Se.44 1834] 414. Spec. Gen. Labiat. Benth., Tre)A ry rc mr cd rs7 8.1868 386. 7: Arts Acad. Amer. Proc. Gray, A. (Torrey) Acanthomintha (Benth.) A Gray) (A. rqe,Nt faz Ege rnl v I .25 1896] 295. A. III iv. Prantl] & [Engler Pflanz. Nat. Briquet, et. l at.30 1849 330. Hartw. Pl. Benth., Tre)Biut a.Pln.[nlr&Pat]4 b.3:25 1896] 295. 3a: Abt. 4, Prantl] & [Engler Pflanz. Nat. Briquet, (Torrey) [ ee,Grefoa2:26 1872] 226. 21: Gartenflora Regel, Eupogogyne .T oel rc ai.Aa.Si 0 1.1931] 116. 20: Sci. Acad. Calif. Proc. Howell, T. J. .2 .T oel rc ai.Aa.Si 0 1.1931] 117. 20: Sci. Acad. Calif. Proc. Howell, T. J. .T oel rc ai.Aa.Si 0 1.1931] 116. 20: Sci. Acad. Calif. Proc. Howell, T. J. rqe,Nt faz Ege rnl ,At a 0.1896] 304. 3a: Abt. 4, Prantl] & [Engler Pflanz. Nat. Briquet, .Ga,Po.Ae.Aa.At 1 0.1876 100. 11: Arts Acad. Amer. Proc. Gray, A. ory?,i ai.Ri.Rp :13 1856] 123. 4: Rep. Rail. Pacif. in Torrey(?), z. aryednee nClfri 2–0 curne threatened); occurrences (20–80% California in endangered Fairly = .THwl,Po.Clf cd c.2:15 1931] 125. 20: Sci. Acad. Calif. Proc. T.Howell, J. t. Acanthomintha rqe,Nt faz Ege rnl ,At a 9.1896] 295. 3a: Abt. 4, Prantl] & [Engler Pflanz. Nat. Briquet, pce ssse to sister as species sn T aaaoe rt clade a to or alone, data ITS using ] rqe,Nt faz Ege rnl ,At a 9.1896] 295. 3a: Abt. 4, Prantl] & [Engler Pflanz. Nat. Briquet, nteraayi.Terphylogeny Their analysis. their in ) Pogogyne and .T oel rc ai.Aa.Si 01518 1931] 20:105–128. Sci. Acad. Calif. Proc. Howell, T. J. a itrt ld composed clade a to sister was pce [ species Pogogyne Pogogyne .douglasii P. oadlavillosa Monardella † oass subgroupings assess to , Taxon yefrgenus. for Type = CNPS pce plus species .duttonii A. . ( .douglasii P. aionaNtv ln oit netr Listing; Inventory Society Plant Native California = .1 itrete oa to either sister eiul nagrdi aiona(vr8%o curne hetndhg ereand degree threatened/high occurrences of 80% (over California in endangered Seriously = Acanthomintha Monardella Pogogyne Bentham, (Abrams) RSMDEXTINCT PRESUMED Pogogyne Pogogyne .I this In ). ihsnnm nbakt n niomna itn hw fo NS2012). CNPS (from shown listing environmental and brackets in synonyms with , ntieMnha fDe n ysa(01 n utieMenthinae subtribe and (2011) are Sytsma studied (2012). and Sytsma taxa and All Drew Drew tree. of of the Mentheae root tribe to outgroups in distant most as included pce n ape of samples and species distant franciscana [ rmr .5lrvrepie,12 MgCl l 1.25 forward primer, l 1.25 reverse DNA, diluted l l l 1 1.25 polymerase, 25 TAQ primer, were l volumes 0.125 of reaction Technol- consisted All AE (Life and California). Cycler of Thermal Carlsbad, l, 2720 Corporation, 100 Biosystems was ogies was Applied of Amplification DNA. an instead of protocol in l, concentration out higher mini 50 carried a yield with plant to eluting order in DNeasy of buffer exception the the of with (Doyle California). followed protocol protocol Valencia, (Qiagen, CTAB manufacturer’s kits the mini plant of The DNeasy version or modified 1987) Doyle a and either using tissue leaf aaClfri,Mxc,wsicue.I re ots h monophyly the test to ilicifolia order A. In included. was of Mexico, California, of Baja species specimen new possible a one from addition, tentatively zizyphoroides were In which as 2006), past. (Meinke identified Oregon in the study conservation ous in described seven, of the infraspecies of each of of our individuals douglasii in P. of two used be least species could samples At that extant 34 1). data sequence these, (Appendix yielded analysis Of species) specimens. verified 16 herbarium (of vouchered from obtained 8 fttlrato) . gfe ufr n 435lH l 14.375 and buffer, Mg-free l 2.5 reaction), total of (8% ln pcfcpies T5 n T21,dsge yKnWurdack Ken by designed ITS241r, and ITS5a primers, specific plant vlto,boegahchsoy pce eiiain and delimitation, times. species diversification history, biogeographic evolution, .lanceolata M. xrcin,Apiiain n Sequencing— and Amplification, Extractions, ao Sampling— Taxon Pogogyne h T eino h ula iooa N a mlfe sn the using amplified was DNA ribosomal nuclear the of region ITS The Pogogyne ) Hedeoma A ry .Ga,and Gray, A. Gray) (A. Emr oes] n n pce n apeo h more the of sample and species one and Jokerst], (Elmer) eeicue,fo ein htcrepne oterange the to corresponded that regions from included, were he pce n ape of samples and species three , a nlddfo aho h he il ie rmaprevi- a from sites field three the of each from included was oepplto;seApni ;Fg ) lo n specimen one Also, 3). Fig. 1; Appendix see population; (one Pogogyne 1B .Gray, A. [ .nana H. ae hetnd rednee nClfri and California in endangered or threatened, Rare, = Pogogyne aeil n Methods and Materials CBR oa f5 ape eecletdi h il or field the in collected were samples 59 of total A CA Mentha aff. .macrantha M. Tre)Biut eeicue.Fnly two Finally, included. were Briquet] (Torrey) osdrdbtrejected; but Considered = CBR 4.2 B1C FE CE 1B.1 B1C FE CE 1B.1 B2CE 1B.2 NSC Fed CA CNPS CBR aionaEdnee pce c (CESA) Act Species Endangered California = eeicue Fg ) i pcmn of specimens Six 3). (Fig. included were serpylloides POGOGYNE Pogogyne ( .arvensis M. .obovata A. .Ga,and Gray, A. (labeled toppltos and populations) (two Acanthomintha .and L. 2 esn,treof three Jepson], .5lDT,2lDMSO l 2 DNTP, l 1.25 , N a xrce from extracted was DNA “P. .pulegium M. p” rmnorthern from sp.”) .villosa M. CE 2 0. [ .lanceolata, A. California = Monardella . were L.) P. subsp. 783 aff. Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. tprpey(arrow). periphery at B. calyces. clareana, and bracts within included 8 YTMTCBTN Vlm 38 [Volume BOTANY SYSTEMATIC 784 E. ooyenudiuscula, Pogogyne Fig. .Poorpsof Photographs 1. aigsltr lwr naiso nlrsec rcs D. bracts. inflorescence of axils in flowers solitary having hwn w o or etl tmn n uecn tl,telte hrceitco eu.F enlpo containing pool Vernal F. genus. of characteristic latter the style, pubescent and stamens fertile four) (of two showing Pogogyne pce n aia.A–B. habitat. and species ooyezizyphoroides Pogogyne nlrsec,as ihicue lwr.C–F. flowers. included with also inflorescence, Pogogyne ooyeabramsii, Pogogyne subgenus Hedeomoides. lowt oiayfoesi rc xl.Nt aye n bracts. and calyces Note axils. in flowers solitary with also A. ooyeserpylloides Pogogyne Pogogyne subgenus nlrsec,soigflowers showing inflorescence, Pogogyne. C. .abramsii P. Pogogyne Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. zizyphoroides F. B. nta etn tpo 4Cfr4mnts )3 ylso 4Cfr4 sec, 45 for 94˚C of cycles 35 2) an amplified minutes; 1) 4 was conditions: for region 94˚C (PCR) gene of reaction This step chain genus. heating polymerase initial the following within the utility using no or little of rmr okdesl o aaotieo h genus the of ITS outside widely-used taxa more for While easily 2006). worked Kress primers and (Prince Smithsonian the of OF SYSTEMATICS PHYLOGENETIC SIMPSON: AND SILVEIRA 2013] ooyezizyphoroides Pogogyne Fig. ooyeclareana Pogogyne .Dsrbto ag aso h ih pce of species eight the of maps range Distribution 2. fti study. this of ” .C. ag eiie ydse ieare line dashed by delimited Range . .douglasii P. –.Subgenus D–F. . Pogogyne Hedeomoides Pogogyne Pogogyne hywere they , .D. –.Subgenus A–C. . ooyefloribunda Pogogyne aao neti dniy(ene20) nldn “ including 2006), (Meinke identity uncertain of taxa adi n aks(98.Teohrpie sdwsESB 5´ a ETS-B, was used primer other The by (1998). developed 18S-E, Markos primer specific and for plant Baldwin the 72˚C using of amplified extension was an DNA 3) and minutes; 2006). 2 Kress for and (Prince 72˚C minutes and 6 sec, 45 for 58˚C h xenltasrbdsae ES eino h ula ribosomal nuclear the of region (ETS) spacer transcribed external The Pogogyne .E. ooyeserpylloides Pogogyne .A. ooyeabramsii Pogogyne POGOGYNE and , .tenuiflora P. .nudiuscula P. P aff. . serpylloides peue extinct). (presumed ,and ”and“ Pogogyne P aff. . 785 sp. Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. 8 YTMTCBTN Vlm 38 [Volume BOTANY SYSTEMATIC 786 Fig. .Seii oaiiso ape of samples of localities Specific 3. Pogogyne sdi hsaayi,wt ebru ceso ubr seApni )indicated. 1) Appendix (see numbers accession herbarium with analysis, this in used Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. ooyezizyphoroides Pogogyne zizyphoroides Pogogyne Pogogyne serpylloides Pogogyne nudiuscula Pogogyne nudiuscula Pogogyne floribunda Pogogyne floribunda Pogogyne douglasii Pogogyne douglasii Pogogyne douglasii Pogogyne douglasii Pogogyne douglasii Pogogyne clareana Pogogyne clareana Pogogyne Pogogyne Pogogyne Pogogyne abramsii Pogogyne villosa Monardella lanceolata Monardella pulegium Mentha arvensis Mentha nana Hedeoma obovata Acanthomintha 72˚C of extension an 3) and OF SYSTEMATICS minute; PHYLOGENETIC SIMPSON: primers 1 AND SILVEIRA 61˚C for minute, 1 72˚C for 94˚C and minutes. initial of 5 cycles an for minute, 35 1) 2) 1 minutes; conditions: 5 gene PCR for for This 94˚C following of 2002). the step Olmstead using heating and amplified (Beardsley was Phrymaceae region in used primer 2013] aae nPU*401 Sofr 02,wt ,0 admaddition random 1,000 with 2002), (Swofford each 4.0b10 on PAUP* performed was in search dataset parsimony heuristic A analyses. Bayesian data missing ambiguous, absent, as either scored were were they gaps. cells because or matrix matrix data data combined of the 23.6% in only, of ITS total A 1) 12775). 4.08 constructed: were MacClade matrices rps16 data in 3) Five only, manually ETS 2010). 2) performed al. et GenBank be to uploaded (Benson were sequences could All 2005). Maddison alignment and (Maddison sequences, final among similarity the a of Because Michigan). Arbor, Ann Codes, Diego San the at Facility. Biosystems) Core (Applied MicroChemical University sequencer State automated higher a 377 recover on ABI to cycle-sequenced order were an in products l PCR 50 Cleaned of product. instead of l concentration 30 with of done elution being final product the the the of products, exception the PCR kits with purify used to purification was kit protocol microcentrifuge PCR the using QIAquick In using California). Valencia, cleaned (Qiagen, used temperature were extension ones were a successful 2007). an to and temperatures al. 3) et up annealing and (Shaw ramping for published minutes; five previously 59˚C temperature 4 as The sec, annealing 62˚C for 12 of minutes. the 65˚C for for 5 simulate 56˚C and 95˚C sec, of to for sec, cycles 12 12 65˚C 30 for for 2) 53˚C of 62˚C minutes, minute, 5 sec, 1 for for 12 conditions: 80˚C 50˚C PCR of minute, following step 1 the heating using initial an amplified 1) was region spacer plast The hlgne eeifre rmec aastuigprioyand parsimony using set data each from inferred were Phylogenies (Gene 4.7 v. Sequencher using edited and aligned were Sequences electrophoresis gel agarose using visualized were products PCR All Table h ia,ainddt eesbitdt reAE(td number (study TreeBASE to submitted were data aligned final, The . trnQ-rps16 trnQ p S 723Esre 7.7 4 Exserted 679273 RSA sp. aff. aff. aff. .Caatr n hrce ttsue ncaatreouinsuyfrseieso l aaicue ntemlclranalysis. molecular the in included taxa all of specimens for study evolution character in used states character and Characters 2. zizyphoroides serpylloides serpylloides (UUG) trnQ-rps16 ao ceso ool eaielnt etl tmn nlrsec it (mm) width Inflorescence stamens Fertile length relative Corolla # Accession Taxon subsp. pcrrgo fclrpatDAwsapiidusing amplified was DNA chloroplast of region spacer and rps16 franciscana ny )ISadES n )IS T,and ETS, ITS, 5) and ETS, and ITS 4) only, 1 ie ySa ta.(07.Ti chloro- This (2007). al. et Shaw by cited x1, DU127Icue 16.4 17.0 17.4 10.2 8.5 13.8 11.8 12.1 8.4 9.1 2 2 11.8 26.4 2 22.0 14.7 2 18.3 2 18.2 4 6.6 4 6.0 4 Included 7.9 2 Included 2 11.3 Included 2 4 Included 4 4 Included Exserted 4 19277 Exserted SDSU 4 104333 Exserted JEPS 4 101601 Included JEPS 4 Included 4 19288 Included SDSU 2 119413 Exserted SBBG 19271 Exserted SDSU 19274 Exserted SDSU 19269 Exserted SDSU 551272 Exserted RSA 1609361 Exserted UC 19279 Exserted SDSU 19290 SDSU Exserted 19285 Included SDSU 19276 SDSU 19273 SDSU 695764 RSA 100839 JEPS 19284 SDSU 111196 SBBG 18438 SDSU DU147Icue 9.3 2 Included 18437 SDSU DU146Icue 8.8 2 Included 18436 SDSU DU120Esre 9.4 12.5 14.5 17.1 14.0 13.8 13.3 12.2 7.2 22.0 4 22.6 4 4 4 4 4 4 2 4 Exserted 4 Exserted 2 Exserted Exserted Exserted Exserted Exserted 19270 Exserted SDSU 19268 Exserted SDSU 17279 Exserted SDSU 16912 Exserted SDSU 16814 SDSU 108833 JEPS 1862058 UC 701934 RSA 593773 RSA 17310 SDSU 12198 SDSU trnQ- aabcueMsut osntspotlklho eosrcinfor reconstruction likelihood support not width does data. number inflorescence continuous Mesquite stamen the for fertile because used and data were was optimization length reconstructions parsimony relative 4.08 Only likelihood corolla data. using version and the optimizations for assumption Mesquite, parsimony implemented states in Both unordered 2010). performed the Maddison char- were All and data. analyses (Maddison continuous as evolution treated were acter and measurements mm study; 6.0–26.4 the was in from used included present ranged specimens width the stamens Inflorescence for corollas, stamens. fertile not four but protrude of bracts, versus not stamens number did two The was as corolla calyces. coded the and if the bracts “included” beyond well as beyond a protruded and length, corolla relative calyces the corolla and if For bracts “exserted” binary. as and coded discrete was as species num- the stamen coded in fertile were features and data diagnostic length ber corolla as relative used for been inflo- states have Character and group. which number, of stamen all fertile length, width, Charac-rescence corolla 2). relative (Table were analysis traced this ters in from used the obtained specimens were within of taxon observation each taxa personal for differentiate states Character to examined. were used genus 2) (Table characters Morphological in evolution and 1995) ETS, (ITS, al. data catenated et (Farris 2002). test (Swofford v4.0b10 (ILD) PAUP* difference in implemented retained. length as were incongruence dis- stationarity the were and reaching using ETS, 30% those ITS, the only first between and The Congruence and burn-in, generations. (Ronquist as 100 carded 3.1 10,000,000 every for Bayesian version performed sampling data. were 2004). generations, MrBayes analyses (Nylander Bayesian missing using All inference 2003). Bayesian as Huelsenbeck performed evolution for of were data treated model the the analyses determine fit were to best used Indels that was 2.3 branch 1985). version assess MrModeltest to replicates (Felsenstein replicate bootstrap per 500 additions support random of 10 total with A performed swapping. were branch TBR and replicates h retplg eutn rmteBysa nlsso h con- the of analysis Bayesian the from resulting topology tree The Pogogyne eas thdtebs ld upr sebelow). (see support clade best the had it because trnQ-rps16 POGOGYNE trnQ-rps16 a sdfrassigcharacter assessing for used was ) aast a assessed was sets data 787 Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. nnomtv,ad8 eebt aibeadparsimony parsimony and but variable variable both were were 75 81 constant, and uninformative, were sites 300 eswithin charac- ters informative variable parsimony and were variable (eight 35 both informative were parsimony constant, 32 were and sites uninformative, parsimony 778 but length), in bp (845 ohvral n asmn nomtv sxparsimony (six informative within parsimony characters were informative and 72 and variable uninformative, parsimony both but variable were chloroplast 48 the For obtained. were region sequences ETS unique and sequences, 28 ITS unique 21 haplotypes, DNA chloroplast 28 8 YTMTCBTN Vlm 38 [Volume BOTANY SYSTEMATIC 788 ih*crepn otoeta eeas on ntesrc osnu reo h ersi asmn search. distinguished parsimony Nodes heuristic analysis. the of parsimony tree heuristic consensus separate strict the from in obtained found 65% also were above that values those bootstrap to correspond to * correspond with below values and probabilities er o o-oignDA(a ta.20) h nlsswsrnfor run was analysis million The per 2006). al. 0.1–0.8% generations. et and million non- (Kay 2000) 10 for nrDNA Muse non-coding years 1987; for million al. years used, per et (Wolfe 0.1–0.3% was for cpDNA of 0.2% coding used model angiosperms of rate in we GTR+G mean rates a Because divergence A with used, RESULTS). zero. was clock (see the at strict a set MrModeltest rate, standardized respectively, from a were included, or determined dates excluded Tip as stem priors. the the with as at monophyletic divergence associated of as extant (and times set of 1.7.1 estimate node crown to version and 2012) and BEAST stem al. data software et The the complex. Drummond the into software; in input divergence were lineage of tree time clades the and visualize to tree, Bayesian noted. data were ranges concatenated biogeographic the with corresponding ranges on The study. superposed this in were included collections from and literature primary rmte3 ape o hc sfldt eederived, were data useful which for samples 34 the From Fig. analysis Bayesian concatenated the from generated was phylogram A the from determined was taxon each of distribution geographic The trnQ-rsS16 .Te rmBysa nlsso octntddt IS T,and ETS, (ITS, data concatenated of analysis Bayesian from Tree 4. trnQ-rps16 Pogogyne aaad04%frteISdt,teebsdo average on based these data, ITS the for 0.45% and data 90b nlnt) 7 ie eeconstant, were sites 870 length), in bp (990 .FrteESrgo 46b nlength), in bp (456 region ETS the For ). Results Pogogyne Pogogyne pce.All species. .FrteISregion ITS the For ). Pogogyne aawere taxa eecnaeae,ol hs aaaerpre ee(i.4). (Fig. here reported and are ETS, data ITS, these which only concatenated, in were tree, evidence total the for and ITS, ETS, 5) nodes); (4 tree rps16 ITS plus ETS 4) nodes); (3 1) sapseirpoaiiyo 0)wti the within 90%) of probability define posterior we a (which well-supported as were nodes of of numbers monophyletic ing analyses strongly-supported, Bayesian a partition. each Pogogyne resolved with data dataset analyses, separate Bayesian each a the in in used marker was evolution (ETS, data of of set and each for ITS, evolution of model appropriate the weak. generally was genus the within clades nomtv 1 asmn nomtv hrceswithin characters informative parsimony Pogogyne (12 informative 5 tp og asmn nlsso l aaesresolved were datasets that all of monophyletic trees analyses strongly-supported, parsimonious Parsimony a long. equally steps 32,075 and 453 ETS, used in that ITS, analysis containing resulted parsimony set The data long. the steps trees 312 parsimonious equally were 50,000 that also of sequences together maximum ETS the and resolved ITS using analyses data The three sets. the among of congruence dif- confirmed total length test (ILD) incongruence ference A The from search. long. resolved parsimony were steps heuristic long, steps the were 76 232 each were trees, that data that ETS 50,000 trees trees resolved resolved resulted 50,000 data ITS 50,000 the in of analysis parsimony of The long. steps maximum 134 the duced trnQ-rps16 roets uigAC eemndta T was G + GTR that determined AIC) (using MrModeltest asmn nlsso the of analyses Parsimony trnQ-rps16 re( oe) ie h uhgetrspotvalues support greater much the Given nodes). (9 tree o h re eie rmteedtst,tefollow- the datasets, these from derived trees the For . .W i o hc o eet nteESregion. ETS the in repeats for check not did We ). trnQ-rps16 .Vle bv rnhscrepn oBysa posterior Bayesian to correspond branches above Values ). re( oe) )ISte 1nd) )EStree ETS 3) node); (1 tree ITS 2) nodes); (0 tree vlae neednl) hsmodel This independently). evaluated trnQ-rps16 Pogogyne trnQ-rps16 hools aapro- data chloroplast u upr for support but , Pogogyne trnQ-rps16 sequences clade: trnQ- Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. 03 IVIAADSMSN HLGNTCSSEAISOF SYSTEMATICS PHYLOGENETIC SIMPSON: AND SILVEIRA 2013] nlrsec it esrmnswr iie nodsrt aeoisadmpe ntete.Dre hdsidct ie . wider indicate shades Darker tree. the on mapped and categories discrete into divided were measurements width inflorescence numb stamen Fertile B. (black). calyces and bracts within Subg. included corollas (black). inconspicuous two versus versus (white) (white) calyces and bracts from exserted corollas aeae T,ES and ETS, ITS, catenated egh hte xetdfo ricue ihnadjacent within included or from exserted corolla whether relative of length, analysis Character reconstruction. state tral Fig. con- the from tree inference Bayesian consensus The Fig. .Prioyotmzto fifoecnewdhcniuu aabsdo refo octntdBysa nlss otnosdt from data Continuous analysis. Bayesian concatenated from tree on based data continuous width inflorescence of optimization Parsimony 6. conspicuous length: corolla Relative A. analysis. Bayesian concatenated from tree on characters of optimization likelihood and Parsimony 5. trnQ-rps16 Hed Subgenus = . aa a sdi ances- in used was data, Hedeomoides . eie 0%poaiiyta h hnehpee nthe on analysis happened likelihood change included the The that to tree. probability exserted 100% the a from on derived once change occurred a optimiza- corollas that Parsimony 5A. determined Fig. in tion shown is calyces, and bracts POGOGYNE r four er: 789 Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. oto nthe four in from shown number, two, stamen is in to reduction four, anal- single likelihood a or and support optimization two ysis parsimony Both whether 5B. Fig. number, in stamen samples fertile all containing of node subgenus the with to used leading branch internal 38 [Volume BOTANY SYSTEMATIC 790 lrsi Province. Floristic nFg .Etmtso h ie fdvrec fndsfrom nodes seen of is divergence analysis are of the same times using analysis the tree the BEAST of the of Estimates inference phylogram 8. A Fig. Bayesian in 7. concatenated Fig. in the shown on ranges bins noted. 11 were into trends divided clear no and 6); continuous (Fig. were data optimi- the parsimony as lineages by zation, assessed the only was in width Inflorescence reductions independent as to well as 96.7%), hsi h is oeua nlsso h niegnsand genus using analysis entire The the work. genetic of future analysis for baseline molecular a creates first our the to is of This contributed relationships phylogenetic has the of here understanding presented study phylogenetic the for Ma) 1.0–7.1 the = for were Ma) CI data 0.4–1.4 95% ITS 0.1; the ± Pogogyne error from (standard estimates Ma divergence 5.1 of times the for ble Ma) 1.1–3.0 Ma) = 5.1–10.5 CI = 95% CI 95% the ±0.05; for error (standard (Ma) ago years Fig. uepstoso oecut,sae rfoitcregion floristic or state, county, some of Superpositions hlgntcAaye n Classification— and Analyses Phylogenetic cnhmnh ilicifolia Acanthomintha .Lclte fslc ldssproe nte rmcnaeae aeinaayi.GF ra ai lrsi rvne F California = CFP Province; Floristic Basin Great = GBFP analysis. Bayesian concatenated from tree on superposed clades select of Localities 7. Pogogyne tmnd n . sadr ro .2 5 I= CI 95% 0.02; ± error (standard 0.9 and node stem tmnd n . a(tnaderr±0.03; error (standard Ma 1.9 and node stem Hedeomoides Pogogyne Hedeomoides Discussion n to and rw oe(i.8). (Fig. node crown Pogogyne trnQ-rps16 ld of clade Fg A.Caatrtracing Character 5A). (Fig. eem nana Hedeoma rw oe Compara- node. crown Pogogyne aawr . million 7.7 were data vrl,the Overall, (likelihood Fg 5B). (Fig. Pogogyne . .zizyphoroides P. ape n one and samples h octntdIS T,and ETS, ITS, concatenated the reo h asmn nlss;teetresmlsand samples three these analysis); other consensus strict parsimony two the the the in found also of clade tree this 0.97, = also (PP clade was parsi- and heuristic 62) the = of Within tree BS search. consensus 1; mony strict = the (PP in support recovered moderate to Subgenus 4). high Fig. 100; = BS among Pogogyne relationships the resolve genera. to three needed these be genus will the mation to relatives close likely P .2 S=5) ihnti clade, this support Within strict 50). low the = to and BS Pogogyne moderate 0.82, tree had = Bayesian (PP tree, the parsimony both consensus in recovered The 4). heu- clade, (Fig. (BS) the bootstrap 70% of had tree that values nodes consensus nodes 10 strict 17 and search, the 0.90, parsimony in (PP) ristic found probability also posterior were a that with nodes 15 of upre ld P .9 S=7;Fg ) h nyother only The 4). Fig. within 73; = clade BS 0.99; well-supported = (PP clade supported Sytsma and these Drew of that All of markers. indicate those molecular different to analyses used similar who are (2012), results ferent grouping in Monardella differs parsimony the shown) However, (not 0.52). = analysis (PP analysis Bayesian the in eut rmtecnaeae aeinaayi indicate analysis Bayesian concatenated the from Results sasrnl-upre oohltcgop(P=1; = (PP group monophyletic strongly-supported a as omagopsse to sister group a form oehrwt Svleo 7.Teetodif- two These 57%. of value BS a with together .floribunda P. ape n n eaieynab sample nearby relatively one and samples .floribunda P. Hedeomoides Acanthomintha Acanthomintha Hedeomoides pcmn locmrs well- a comprise also specimens pcmnfr well-supported a form specimen trnQ-rps16 Monardella Hedeomoides ,thetwo Pogogyne, smnpyei with monophyletic is - and Monardella Acanthomintha Acanthomintha aasosatotal a shows data ihlwsupport low with P. u oeinfor- more but Monardella r h three the are aff. serpylloides - Pogogyne and and are Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. P .7 S=6) h he ape of samples three The support 69). strong = BS somewhat 0.97; with = (PP monophyletic is subgenus 0.51, Within samples) well-supported. 4). (PP are Fig. poor see clades conclu- is 50; branches not basal < is for BS subgenus support the that of given sive paraphyly the but analyses, pcmn(DU125 olce nLk onyada and County Lake in a collected new with possibly 19285) along clade, (SDSU this localities of (see specimen samples County The 1; 3). Obispo = Fig. Luis (PP at of support San specimen strong adjacent with one from support sister to lected is strong 78) and = 100) very = BS 4). 4). with BS 1; (Fig. monophyletic (Fig. = support (PP is support 50) samples) < strong (two (BS lacks low pattern to abramsii 0.86) having clade Pogogyne = this (PP below), see moderate California; County, of samples Diego two San including clade a of part oe,telte ersnigtetm fdvrec fextant of divergence of time the representing latter the nodes, upr nteBysa nlss(P=09,as on in found also strong 0.92, has = that (PP clade analysis Bayesian a the form in Mexico, support California, Baja ern found 4). also Fig. analysis; parsimony clade the this of consensus 0.84, strict the = in (PP clade well-supported ately e oaiya i.3.Teefu ape lsteohrsample other the plus samples of four These 3). Fig. at locality see of OF SYSTEMATICS PHYLOGENETIC SIMPSON: AND SILVEIRA 2013] Subgenus Fig. .serpylloides P. .serpylloides P. .Pyormfo octntdBysa nlss hw eo r ES siae ftetm foii fthe of origin of time the of estimates BEAST are below Shown analysis. Bayesian concatenated from Phylogram 8. Pogogyne Pogogyne n htof that and rmLk ony aiona(DU19288; (SDSU California County, Lake from sprpyei nalaaye,btthis but analyses, all in paraphyletic is ncnrs,i o oohltci our in monophyletic not is contrast, in , p RA697)cletdi north- in collected 679273) (RSA sp. P. ooyenudiuscula Pogogyne aff. .douglasii P. zizyphoroides Pogogyne .abramsii, P. SS 90)col- 19309) (SDSU ooyeclareana Pogogyne .nudiuscula P. omamoder- a form fo three (from nyafew a only .douglasii P. Pogogyne alfrom (all are taxa. o he diinlrdcin nsae ubri the in number stamen ilicifolia Acanthomintha in in evidence one reversals) reductions provides complex, additional 2–3 study This three with Menthinae. for (along subtribe number their in stamen of species in one reductions only which in ( (2012), Drew Sytsma by Lamiaceae and the and of two analysis branch phylogenetic to internal higher-level subgenus four same for apomorphies from the clear on are and happened corolla conspicuous stamens inconspicuous a fertile analysis. an from change to likelihood the corolla that and suggest optimization analyses based These stamens, parsimony fertile both four and on flowers exserted had clearly 50; < (BS value bootstrap 4). low Fig. a see but analysis), parsimony the olnto nsubgenus self- in to outcrossing from shift possible a of because correlated iseg19;TopadLog19) oeo h species the of None 1998). subgenus Leong within and and Thorp (Spencer spe- self-pollinating 1998; plant are Riesberg pool several (Schiller that vernal outcrossing noted other have of predominately cies studies However, be 2000). al. to et found been has in .douglasii P. hrce Evolution— Character euto nbt ool ieadsae ubrmybe may number stamen and size corolla both in Reduction Pogogyne ih4saes a nldd upre three supported included, was stamens) 4 with , aeol encnutdon conducted been only have Hedeomoides Pogogyne and POGOGYNE h omnacso of ancestor common The eem nana Hedeoma Hedeomoides n needn eutosin reductions independent and aebe tde ihregard with studied been have Hedeomoides olnto studies Pollination . Pogogyne . .abramsii, P. tmadcrown and stem Fg ) The 5). (Fig. Pogogyne Pogogyne which 791 Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. a encsayt eov hsissue. among this resolve relationships to necessary the be may of Monardella, resolution Acanthomintha, poor present relatively was or the the groups in lost two of later ancestor occurred the common taxa in the two in convergence the by in style either pubsescent a monophyletic, measures consistent (e.g. had width species inflorescence individual of subgenus Some of 6). members some (Fig. with with overlap but and widths, inflorescence variation moderate to small have 9 YTMTCBTN Vlm 38 [Volume BOTANY SYSTEMATIC 792 u hlgntcaayi.Creae ihtemonophyly the genus with the Correlated of an analysis. be phylogenetic to our there expect not would we difference.) although appreciable not width field, specimens, herbarium the (Inflorescence from in measured years. inflores- was study wider wetter this with for with inundation, correlated pool vernal al. cences precipitation of et of plastic amount length environmentally the Silveira and as be such 1993; well factors by to may Jokerst influenced and others feature (e.g. this with keys but combination 2012), in in used species still Inflo- separate 2012). is al. et width Silveira of (e.g. rescence authors subspecies later by the followed larly lumped who douglasii is inflorescence (1943), P. of inference Jepson there This variability with phylogeny. that the known agrees in with suggest correlates trend that analyses or width signal these no of but likely infraspecies 1931), different describe (Howell to used e.g. been samples, different douglasii among P. variability considerable had and in subgenus trends of obvious no Members showed Analysis feature. 6) species. (Fig. this data described continuous a the within of consistent always not to needed be genus. the will within mechanisms manipulations, exclu- reproductive transfer verify pollinator studies, pollen ratio, and reproductive pollen:ovule sion, a of Future with measures a coincide self-pollination. including with could along on number, this, stamen reliance visitors; inflorescence fertile insect corollas the in to of that reduction visible calyces possible less and them is bracts make it the within but inserted mechanism, pollination to xml fapbsetsyeaoggnr lsl related closely genera among style to pubescent a of example inlybe sdt characterize to in used been tionally very grasslands, pool. within vernal miniature a swales to consist similar can inundated species us this periodically of of of habitat one the by that observation confirms (Silveira) personal However, grasslands. addition, in of In habitat aquatic. the is Regardless, vernal comm.). or pers. (Elizabeth this pools through Painter, running However, vernal stream intermittent in pools. an found with vernal pools be to occasionally can opposed species as streams basis. dated genetic a in have occurred to presumed is Within that pool) vernal feature (mostly a wet a habitat, in reproduce and survive to ability te etrso hrce vlto a entdfrom noted be can evolution character of features Other were and varied data width inflorescence measured The ial,w oeta uecn tl,wihhstradi- has which style, pubescent a that note we Finally, cnhmnh lanceolata Acanthomintha Pogogyne .zizyphoroides, P. Pogogyne Fg ) nlrsec it safaueta has that feature a is width Inflorescence 6). (Fig. nhsteteto h ru,atetetsimi- treatment a group, the of treatment his in Because . .clareana P. Pogogyne cnhmnh lcflaA obovata ilicifolia-A. Acanthomintha lgteouinr hf a have may shift evolutionary slight a , ihamdrt it)wieothers while width) moderate a with samjreooia pmrh:the apomorphy: ecological major a is hc rw nproial inun- periodically in grows which , Pogogyne .serpylloides P. hsi h nyohrknown other only the is This . Pogogyne and .clareana P. Pogogyne, ssrnl-upre as strongly-supported is Pogogyne and Hedeomoides sctda occurring as cited is ihasalwidth, small a with Acanthomintha ute studies further sas found also is , ld.Given clade. .douglasii P. .clareana P. eddto tended Pogogyne and pcmn of specimens ercsuymyb eddt eeiemorphological redefine to needed be of boundaries may morpho- species in-depth study an so delimitations, metric not species do current that the discovered fit been have populations New features. ooti T aafrti ape(pedx1;tu,the thus, 1); (Appendix on sample based this for is data analysis ITS obtain to a o eoee ntecaewt a ig onyspec- with along County clade Diego sister San a with 679273) within clade but (RSA imens the in Mexico recovered California, axil not Baja was leaf northern per of in specimen flowers single pri- collected The of differing 2012). number al. relatives, et and close (Silveira pubescence be species in to proximate thought marily geographically been two long These have 7). (Fig. clade nudiuscula P. usOip ony pcmn of Specimens San adjacent County. of region Obispo nearby Luis a from collected 19309) (SDSU orsodnecnb aebtenvroscae and clades various of subgenus between some Within ranges. made but geographic no be performed, regions, can was correspondence biogeographic analysis delimiting biogeography detailed with difficulty and ocp eeue eg e eQerz19) hr would there 1998), species Queiroz phylogenetic De of see (e.g. several type used some of were If concept biology evaluated. reproduction be cannot the of Pogogyne studies no tually differ- the of describe species to used ent been (Cronquist traditionally has concept 1988) species 1978, morphological A analyses. these genus the within boundaries Pogogyne. species of elucidation for study ssrnl-upre smnpyei P .) hssingle this specimens 1.0); of = three (PS sample monophyletic of as consisting strongly-supported is group a of clade, this includes within as BS<50) identified 0.84, latter, of = This samples 7). all (PP Fig. clade (CFP; Province moderately-supported Floristic subgenus California the of in other clade all other (like found the of identified specimens members two the and of California, as specimens of (GBFP; GBFP clade, Province the all this in Floristic includes In 73) Basin 7). = Great BS Fig. the 0.99, = from (P specimens One well-supported well-supported. is that to clade moderately- each clades, correlated investigation. further needs as and taxon identified as formerly (represented specimen, Mexican hsnro ein sdsusderir h itrtaxon sister the earlier, discussed As of region. narrow this ahohrscoetrltv ihgo upr Fg ) this that 7); given (Fig. surprising support not good is with result relative closest other’s each 143 rmSnLi bsoCut Fg ) aty a Lastly, 7). genus (Fig. as County identified zizyphoroides tentatively Obispo Oregon, from Luis specimen single San from 119413) pcmn hni h other the is the than to specimens proximate geographically more much Biogeography— Subgenus pce Delimitation— Species .clareana P. .clareana P. .zizyphoroides P. P. Hedeomoides aff. pce,abooia pce ocp My,1963) (Mayr, concept species biological a species, oeifrne nseistpscnb aefrom made be can types species on inferences Some serpylloides .serpylloides P. l rmSnDeoCut,Clfri,fr a form California, County, Diego San from all , fMnee onywsa was County Monterey of SS 83) ssse oters ftesub- the of rest the to sister is 18438), (SDSU olce nMnee onywr on obe to found were County Monterey in collected Hedeomoides .douglasii P. .serpylloides P. P. Pogogyne P. ie u eaieysalsml size sample small relatively our Given aff. ld rmteCFP. the from clade p nti td) ol ersn new a represent could study), this in sp. n n of one and Pogogyne Pogogyne trnQ-rps16 r rmteGF fOeo.All Oregon. of GBFP the from are zizyphoroides sn n rmr morphological more or one using , Fg ) oee,w eeunable were we However, 7). (Fig. u nlsscntttsabaseline a constitutes analysis Our rmLk ony aionais California County, Lake from xiie w biogeographically two exhibited , .zizyphoroides P. ape sdi hsanalysis) this in used samples eas hr aebe vir- been have there Because . .serpylloides P. .serpylloides P. n T aaaoe This alone. data ETS and .clareana P. .floribunda P. Pogogyne, Fg ) Interestingly, 7). (Fig. ooyeabramsii Pogogyne .douglasii P. .clareana P. Pogogyne n specimens and , Hedeomoides .zizyphoroides P. ape(SBBG sample srsrce to restricted is SS 19288) (SDSU .nudiuscula P. ohsamples both r found are specimen n two and species P. and aff. are Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. hr r eysalbac egh ihnthe within lengths branch small very long calibra- are relatively there fossil the to some contrast using In tions. a Lamiaceae, only the the of analyzed from representatives who Ma) (2012), 4–7 Sytsma ca. single = and CI Drew (95% of ago analyses years million 6 imately ru Fg ) seilywe oprdtolimited). compared when especially and 8), (Fig. group naeaeo .–. ilo er g,atog with date although This ago, 8). (Fig. years the with intervals well million corresponds confidence 5.1–7.7 wide and of considerably Drew arisen average have also to respectively) an (see data, nrDNA ITS another and along chloroplast to The occurring taxon 2012). changes Sytsma one of number patristic from of total terms lineages the in relatives i.e. close its distance, from separated well is that illustrates 8) (Fig. set to interrela- needed their genus. be the and in may delimitations tionships species 2008) recon- understand Edwards tree whole better species (see perhaps of or methods use 2009), micro- struction potential al. markers, and et nuclear sequences, Edwards better genome copy in to (as single data needed as satellite are such of data, studies genetic evolution further the group, understand the of frame- evolutionary the work resolved and variation some showed speciation. incomplete of products the represent .clareana P. oietf eaiet ecie pce.Atog h taxa the difficult Although are species. that described Oregon to subgenus from of relative samples identify of to supported lineage existence be may incomplete the notion by This of clearly introgression. not genetic because are or sorting perhaps which of distinct, lineages the genetically rel clade, may evolved used resolve recently data a the fully that clear to is Hedeomoides it unable case, any be In analysis. our in but support. paraphyletic, strong with zizyphoroides monophyletic as recovered paraphyly observed sorting. lineage the plete us for morphological slight by explanations of undergone synonyms Other line- have of which as changes. assemblage of paraphyletic treated some a past, ages, represent may the 1), in (Table of described varieties been and subspecies, species, usqetydvre rmec te,a xml of explain possibly example could of an samples This some other, why speciation. each isolate from and peripheral isolated become diverged that lineages wide-ranging this subsequently separate By to the led 4). have (Fig. of could support closely populations good of more with are hypothesis, (plesiospecies species, samples other species some to that related paraphyletic given OF species. a 1995), SYSTEMATICS PHYLOGENETIC phylogenetic Olmstead as SIMPSON: AND a qualify SILVEIRA deemed could molecular support be sufficient has well-supported to clade other on no however, based apomorphies; species both phylogenetic for support be 2013] iegneTimes— Divergence study this in used markers molecular the Although oeo h niiulseiswithin species individual the of None .douglasii P. Monardella Pogogyne Acanthomintha hnt te pcmn of specimens other to than n/rta hscmlxo pce represents species of complex this that and/or , Hedeomoides a edtrie snihrmnpyei or monophyletic neither as determined be can nld yrdzto ihohrtx rincom- or taxa other with hybridization include atog h apesz o h atris latter the for size sample the (although pce ( species .serpylloides P. Pogogyne h hlga ftecmie data combined the of phylogram The .douglasii P. .clareana P. so pca oebcuetreo the of three because note special of is r eeal dniibe hycould they identifiable, generally are .douglasii P. Pogogyne Pogogyne sdfntvl non-monophyletic definitively is tmnd setmtd(from estimated is node stem Pogogyne tosiswti subgenus within ationships r oecoeyrltdto related closely more are wt ogse lineage) stem long a (with and tmnd g fapprox- of age node stem u tde numerous studied but ) .douglasii P. Pogogyne .douglasii P. s fadditional of Use . .nudiuscula P. ooyedouglasii Pogogyne Hedeomoides Pogogyne Acanthomintha tmlineage, stem h various The . .douglasii P. hthave that Pogogyne crown being were pmrhcpyilgclaatto ovra olor pool genus. vernal the of pool an members to in of conditions vernal evolution wetland” adaptation “temporary the a with physiological correlated to apomorphic likely adaptation was pool This vernal their habitat. that in following diversification hypothesize rapid ecosystems dates we relatively these many estimates, a Although for underwent rough 1987). present Harden quite before (see are years soils million esti- pool an 0.6–4 with vernal compares of This age ago. years mated aver- million an be 0.9–1.9 to of respectively) age data, nrDNA ITS and chloroplast 8). (Fig. time of period longer a for separated been dad,C . .S ud .M ot,adB ern.20.Using 2009. Herring. B. and Ionta, M. G. Judd, S. W. E., C. systematics molecular Edwards, of theory general and new a Is 2008. V. S. Edwards, Bayesian 2012. Rambaut. A. and Xie, D. Suchard, A. and M. J., biogeography, A. Drummond, Phylogenetics, 2012. Sytsma. J. K. and T. B. Drew, the Phrymaceae: Redefining 2002. Olmstead. G. R. and M. P. Beardsley, external the of utility Phylogenetic 1998. Markos. S. and G. B. Baldwin, Silveira M. to California awarded the scholarships and study. and University this this grants for State for improved Diego Society thank the Plant San greatly We Native thank thank work. comments we specimens molecular also whose Finally, of the paper. We reviewers, loan for for project. anonymous material UC-JEPS this remove three and to for SDSU, permission SD, work and SBBG, Scott field RSA, and and in at work Guilliams herbaria help Matt lab C. in for thank help We McMillan considerable paper. for this Hasenstab-Lehman of draft Kristen earlier an on ments nenlbac egh r osdrbylne nta genus that in in longer than considerably are lengths branch internal four rw .T n .J ysa 01 etn h oohl n placement and procedure monophyly the isolation Testing 2011. DNA Sytsma. J. rapid K. and A T. B. Drew, 1987. Doyle. L. J. and J. J. Doyle, species species, of concept lineage general The 1998. K. Queiroz, De Meutter, De Van F. Louette, G. Stoks, R. Declerck, S. L., Meester, De 1988. A. Cronquist, NS 02 aionaNtv ln oit netr frr and in 3–20 Pp. species? rare a is what of again, Once 1978. inventory A. Cronquist, Society Plant W. Native E. and California Ostell, 2012. J. CNPS. Lipman, J. D. Karsch-Mizrachi, I. A., D. Benson, aigo the of Dating Acknowledgments. ouaingntcdt sato oietf e species: new identify to tool a as data genetic population emerging? 1.7. BEAST the Evolution and and BEAUti with phylogenetics (Lamiaceae). Mentheae tribe Botany the of Journal in evolution staminal 1038–1049. 36: of placement and ETS of Evolution of congruence trees 18S-26SrDNA: of ITS (ETS) spacer transcribed of tissue. leaf fresh of 11–15. quantities small for Press. University Oxford speciation and in unification and 57–75 conceptual A Pp. speciation: recommendations. of terminological process the and criteria, pools and Ponds evolutionary and 2005. 715–725. ecology Brendonck. biology, L. conservation and biology. in systems Michels, model E. as Bie, De T. Garden. Botanic York New York: New Agriculture in nagrdpat oln dto,v–1) http://www.rareplants v8–01a). .cnps.org. edition, (online plants endangered http://www.ncbi.nlm.nih.gov/genbank. 10.1093/nar/gkq1079. GenBank. 2010. Sayers. Botany of Acanthomintha Lepechinia Pogogyne qai osrain aieadfehae ecosystems freshwater and Marine conservation: Aquatic 9 1093–1102. 89: 0 449–463. 10: Evolution d .A abre.Mncar lahl n Osmun. and Allanheld Montclair: Ramberger. A. J. ed. , Mimulus Calycadenia d.D .Hwr n .H elce.NwYork: New Berlocher. H. S. and Howard J. D. eds. , 9 1969–1973. 29: ntetieMnha (Lamiaceae). Mentheae tribe the in h vlto n lsiiaino lwrn plants flowering of classification and evolution The ugsigthat suggesting , Pogogyne 9 933–953. 99: pce eeicue norsuy and study, our in included were species 3 1–19. 63: etakC atGilasfrvlal com- valuable for Guilliams Matt C. thank We rb iuee and Mimuleae, tribe , ieaueCited Literature uli cd Research Acids Nucleic POGOGYNE (Compositae). rw oewsetmtd(from estimated was node crown Acanthomintha oeua hlgntc and Phylogenetics Molecular htceia Bulletin Phytochemical Phryma 9 aaaeisedoi: issue Database 39: nls om:Species forms: Endless ytmtcBotany Systematic oeua Biology Molecular . mrcnJournal American pce have species Biosystematics Pogogyne Conradina American d 2. Ed. . 793 19: 15: Delivered by Publishing Technology to: Michael Simpson IP: 146.244.235.95 on: Fri, 27 Sep 2013 18:36:45 Copyright (c) American Society for Plant Taxonomists. All rights reserved. oqit .adJ .Hesnek 03 rae :Bysa phylogenetic Bayesian 3: MrBayes 2003. Huelsenbeck. P. 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(Polemoniaceae). d .C ika.Bree:Uiest fClfri Press. California of University Berkeley: Hickman. C. J. ed. , ¨ llersjo h hlgntcsseaisof systematics phylogenetic The Pogogyne Pogogyne ooyefloribunda Pogogyne Evolution 0 623–630. 20: 4 747–759. 34: ¨ .G lg,adC ut 95 etn signifi- Testing 1995. Bult. C. and Kluge, G. A. , evr ntdSae elgclSurvey. Geological States United Denver: . Cladistics p 2 in 724 Pp. . p 1–1 in 416–419 Pp. . Pogogyne :36. 6: 4 275–288. 94: 9 783–791. 39: eiso olcrnsqecsi h western the in chronosequences soil of series A . 0 315–319. 10: 2 25–43. 42: rceig fteClfri Academy California the of Proceedings Lmaee,anwseisfrom species new a (Lamiaceae), clg,cnevto,admanage- and conservation, Ecology, clg,cnevto,admanage- and conservation, Ecology, h esnMna:Hge plants Higher Manual: Jepson The aionavegetation California Bioinformatics abig:BlnpPress. Belknap Cambridge: . ooyeabramsii Pogogyne lr fCalifornia of Flora A Wetlands Aliso clg,cnevto,and conservation, Ecology, Pogogyne Pogogyne 3 347–353. 13: eode. ed. ed., second . 9 1572–1574. 19: 0 386–396. 20: p 850–851 Pp. . Dubuque: . (Lamiaceae) (Lamiaceae) Pogogyne ,pr 2. part 3, ,ed. . akr .adK ysa 06 tmnleouini h genus the in evolution Staminal 2006. Sytsma. K. and J. Walker, KC591699; abramsii Jokerst. lanceolata KC591697. macrantha Monardella vary substitution nucleotide of Rates collection 1987. Sharp. P. and a Li, W. H., of K. Wolfe, List 2. contributions: Botanical VI. 1875. S. Watson, C967 C967 KC591700. KC591667, KC591637, L. 4–110 29) C969 C962 KC591692. KC591662, Curran. KC591629, 12198), Berkeley. California, San of = University SDSU = Garden; Santa UC Botanic Jepson Rancho University; Barbara State = = Santa Diego = JEPS RSA SBBG Berkeley); acronyms: Garden; California, Botanic Herbarium Ana of data. University sequence (at ITS GenBank respectively. Herbarium of and sequences, absence accession, trnQ-rps16 = herbarium and ITS, taxon), * ETS, given for a numbers of accession one than more (if wetlands”. “isolated of concept the and pools Vernal 2003. H. P. Zedler, C969 KC591702. KC591669, clareana HL-2753 83) C963 C962 KC591716. KC591682, KC591653, 18438), KC591723. zizyphoroides 10157 KC591721; Ahart KC591687, KC591658, 094,K513,K516,KC591695. KC591665, KC591632, 701934), KC591694. KC591664, obovata Acanthomintha 089,K514,K517,KC591703; KC591670, KC591640, 100839), C962 C962 KC591705; KC591672, KC591642, KC591704; douglasii 2159 York douglasii 10VI93B McMillan KC591708. KC591714. KC591717. KC591683, serpylloides 15VI05A KC591720. KC591686, serpylloides SLO-80 KC591712. KC591679, nudiuscula Pogogyne 2. KC591711; Gray KC591678, A. KC591648, 19269), (SDSU KC591710. KC591677, KC591647, floribunda Pogogyne 2. Jokerst KC591718; KC591684, KC591655, 551272), Appendix cnhmnh ilicifolia Acanthomintha hr 12913 Ahart .W ihm .Bue,W .Bl,adR rdf.Sacramento: Ornduff. R. and Belk, Society. conference F. Plant 1996 W. Native a Bauder, California from D. Witham, Proceedings W. ecosystems: C. pool vernal of ment Lmaee:mlclrpyoeei vdnefrmlil origins multiple lever. staminal for the evidence of phylogenetic molecular (Lamiaceae): rceig fteNtoa cdm fSine USA Sciences of DNAs. Academy National nuclear the and of Proceedings chloroplast, mitochondrial, plant among greatly Sciences them. with and Palmer, upon Edward Dr. notes by made his Island, Guadalupe from plants of Wetlands U 825) C967 C961 KC591690. KC591661, KC591627, 1862058), (UC ipo 2842 Simpson ipo 2866 Simpson SB 143,K515,K518,KC591719; KC591685, KC591656, 119413), (SBBG SS 83) C962 C961 KC591715; KC591681, KC591652, 18436), (SDSU eta 6, Bentham SB 116,K513,K516,KC591701; KC591668, KC591638, 111196), (SBBG .T oel1. Howell T. J. .T oel2. Howell T. J. eta 3. Bentham ooyefloribundaPogogyne ooyezizyphoroidesPogogyne RA656) C963 C963 KC591706; KC591673, KC591643, 695764), (RSA Pogogyne ivia20 Silveira ooyeabramsii Pogogyne .Gray. A. ooyedouglasii Pogogyne oadlavillosa Monardella Tre)A ry2. Gray A. (Torrey) Tre)A ry2. Gray A. (Torrey) ivia8 Silveira JP 033,K515,K518,KC591722; KC591688, KC591659, 104333), (JEPS .Txn olco,cleto ubr apenumber sample number, collection collector, Taxon, 1. 3 597–607. 23: eta 3. Bentham 1 112–121. 11: JP 083,K512,* KC591691. *, KC591628, 108833), (JEPS SS 98) C964 C964 KC591707; KC591674, KC591644, 19285), (SDSU ciln8VI93 McMillan aff. Pogogyne ar 295 Lauri Pogogyne ooyedouglasii Pogogyne Sade)Jkrt3. Jokerst (Standley) ooyeserpylloides Pogogyne SS 77) C965 ,KC591698. *, KC591635, 17279), (SDSU eem nana Hedeoma .Ga 3. Gray A. SS 97) C966 C966 KC591709; KC591676, KC591646, 19279), (SDSU SS 97) C960 C960 KC591713; KC591680, KC591650, 19271), (SDSU ivia18 Silveira SS 71) C960 C963 KC591693. KC591663, KC591630, 17310), (SDSU .Gray. A. zizyphoroides ivia4 Silveira naso Botany of Annals ivia10 Silveira Jepson. ciln9IV93B McMillan rceig fteAeia cdm fArts of Academy American the of Proceedings A ry .Gray. A. Gray) (A. ciln10VI93FMcMillan Sade)Jkrt1. Jokerst (Standley) aff. .T oel2. Howell T. J. SS 61) C963 ,KC591696. *, KC591633, 16814), (SDSU ene15VII05 Meinke p li 191 Elvin sp. ooyenudiuscula Pogogyne ooyeclareana Pogogyne eta 4. Bentham eta subsp. Bentham ar 273 Lauri SS 97) C961 KC591671, KC591641, 19273), (SDSU od8558 Boyd ooyezizyphoroides Pogogyne eta 1. Bentham serpylloides SS 96) C966 KC591666, KC591636, 19268), (SDSU ivia7 Silveira SS 97) C960 KC591689, KC591660, 19277), (SDSU SS 90) C965 KC591675, KC591645, 19309), (SDSU ooyedouglasii Pogogyne Tre)Briquet. (Torrey) Bentham. ooyedouglasii Pogogyne ooyefloribunda Pogogyne etaarvensis Mentha eta 1. Bentham 0:375–391. 100: SS 61) C964 *, KC591634, 16912), (SDSU sad5669 Oswald cnhmnh lanceolata Acanthomintha Tre)A ry1. Gray A. (Torrey) RA697) C961 *, KC591651, 679273), (RSA SS 98) KC591639, 19284), (SDSU SS 97) KC591649, 19274), (SDSU RA537) KC591631, 593773), (RSA alr17136 Taylor ivia9 Silveira Tre)A ry1. Gray A. (Torrey) SS 83) KC591654, 18437), (SDSU ivia5 Silveira SS 98) KC591657, 19288), (SDSU ene15VI05B Meinke atooe 5932Bartholomew ooyenudiuscula Pogogyne esr30IV94 Reiser .T oel1. Howell T. J. franciscana .Ga 1. Gray A. 4 9054–9058. 84: alr17212 Taylor etapulegium Mentha od11450 Boyd SS 19276), (SDSU SS 19270), (SDSU U 1609361), (UC Pogogyne JP 101601), (JEPS L. eta 5. Bentham eta 2. Bentham Monardella eta 2. Bentham owl AC Colwell (Standley) Pogogyne Pogogyne Pogogyne Pogogyne Pogogyne Pogogyne ivia6 Silveira (Elmer) Painter Painter (SDSU Meinke (SDSU Salvia (JEPS (RSA (RSA ,ed. aff.