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1-1-1993 Phylogenetic Analysis of the Zingiberales Based on rbcL Sequences James F. Smith Boise State University

W. John Kress Smithsonian Institution

Elizabeth A. Zimmer Smithsonian Institution

This document was originally published by Biodiversity Heritage Library in Annals of the Missouri Botanical Garden. Copyright restrictions may apply. Image courtesy of Biodiversity Heritage Library. http://biodiversitylibrary.org/page/553107 PHYLOGENETIC ANALYSIS OF J(lfI/(>S F. Srnitli / ·H IV l ohn Krc:}s,3 and Z 3 THE ZING IBERALES BASED I:'/izabeth A. Zimmer • ON rbeL SEQUENCES'

AnSTRAcr

Morphological data have bet:n used previously to construct phylogenies of the eight families of the Zingiberales, one of the most wi dely accepted monophyletic groups of nowcring plants. To provide additional suppon for phylogenetic relationships within Ihe order, and placement of the order among monacots. we present a parsimony analysis of DNA sequences from the chloroplast-encoded gene, rbe L. for 21 species of Zingiberales and proposed relatives. Five analyses with equal, and differential weights were performed. All analyses resulted in the salllC most parsimonious tree for taxa within the Zingiberales and the irnmediate outgroup. The closest sister group to the Zingiberales based on these data is a clade containing Commelinaceae/ Haemodoraceae/ Pontederiaceae. The tree topology within the order based on r/)('l sequence data is different from previous morphological analyses. The order can be divided into two sister groups, one containing the Costaceae and Marantaceae, and the other, the remaining six families . All recognized families are monophyletic with the exception of the Musaceae, which is paraphyletic with the Cannaceae. With trees one and two steps longer than the most parsimonious trees, phylogenetic resolution is rapidly lost. suggesting that the phylogenetic utility of rbe l sequence data for the Zingiberales is limited to interordinal and intrafamilial relationships.

T he Zi ngi berales. a morphologicall y di stinctive classifica tion, including delim it ation and rank of oreler of monocots. arc one of the most widely ac­ these families, has been subject to many changes cepted monophyletic groups of pl ants (Bentham & (reviewed in Kress. 1990). Cladistic analyses of Il ooker. 1883: Petersen. 1889: Schumann. 1900, morphological charal:1ers ha ve greatly improved 1902. 1904: Hutchinson, 1934, 1959, 1973: Na­ the understanding of ph ylogenetit: relat ionships of kai, 1941: Tomlinson, 1962, 1969: Stebbin :-s, 197 k the families (Dahlgren & Rasmussen, 1983: Cronquist, 1978, 1981: Dahlgren & HasllIu ssen, Kress, 1990). Dahlgren & Rasmussen (1983) per­ 1983; Dahlgren et aI. , 198.'): Kress, 1990). Dahl­ fon ned the fir st cladisti c analysis of the Zi ngiberales gren el aL (1985) li sted six apomorphies for the using the eight families listed above and polarized Zi ngiberales: root hair cells short er than other epi­ <.: hara <.: ters using their Commeliniflorae. This anal­ dermal cells. sieve tube plastids containing starch, ysis resulted in a single tree (Fig. I) composed of presence of silica bodies, epigynous fl owers, lack three main clades that included the gi nger group of distincti ve apert ures on the pollen grains. ami (Z ingiberaceae/ Costaceae and rVl arantaceae/ Can­ the occurrence of arill ate seeds. In addition_ the naceac). the banana group (Musaceae/ Heliconi­ herbaceous arborescent stem, distichous phyllo ­ aceae). and the bird -of- paradi se group (Strclit zia­ taxy. large pet iolate leaves wi th blades possessing ceae i Lowiaecae). The relationships among the three transverse ve nation, conspicuolls colorfu l hral:1eate groups remained equi vocal. in fl orescences. and the substit ut ion of olle to five Kress (1990) re-a nalyzed th e data of Dahlgren staminodia for the fer til e stamens a re characters & Ra smussen ( 1983) and performed a separate easil y lI sed to ident ify members of the Zi ngiberales analys is that included ot her characters which we re (Kress, \990). rooted wit h th e Brorne li ales, T his second analysis As currently classified, the order consists of eight resulted in a different cladogram (Fig. 2) from that fami lies (K ress. 1990): Musaceae. Low iaceae. I-Iel­ of Dahlgren & Hastnlt ssen ( 1983). Although the iconi aceae. Strelitziaceac, Zi ngibcraceae, Costa­ ginger group relationship wa s retai ned in bot h. the ceae, Marantaceae, and Cannaceae. SuLordi nal fam il ies of the banana and bird-o f-paradise groups

, We thank YOllngbae Sli h for expert advice on cloning and sequencing. We acknowledge the Scholarly Studies Program of the Smithsonian Institution fo r providing fundi ng and a grant to WJK, the staff of the US Botall ical Carden for cure of li villg coll ectiuns, and G. Zurawski (DNAX) for providing primer sequences, , l aooratory of Molecular Systematics, Museum Support Center, Smithsonian Insti tution, Washington, D.C. 20560, U.S.A. ' Department of Bota ny N HB· 166, Smithsollian Illst itution, \Va shington, D. C. 20560, USA. , Current address: Department of Biology, Boise State University. Boise. Idaho 83725, U.s.A.

A NN. MISSO UIl' BOT. GAllI). 80: 620- 630. 1993. Volume 80. Number 3 Smith et al. 621 1993 Phylogenetic Analysis of Zingiberales

• • • • • • •v • •v v < • V • • • • <• c • • ,- • v • v • •w ~ •w - • < • •0 •, • w •v < -~ v• • < •c - - < • - • •c u• • 0 - , v 0• U ,• • , v • -- ~

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f lt :1 liE [. Cladogram of the Zill~ibl'ralcs frollt Dahlgren & Ra"lIIu!'o"clI ([ 983. their figu re 9 ). we re show n to he paraphylc tit' (K n·:-:--. 19 90). T hese I-I a rbo rne . 1977 ). T hol"ll(' ( 1992). !J(I\\t. \ t·r. 1'1','­

C1 na]vscs• have heen highv l•\' ill forlna tiq' ill t(,rIl lS of ognized thrce .'- eparalt" o rder" ill hi ... "'lIl" 'rord('r f.. tuilia l relationships. vel the ut il i!\' of m a ll \' o f the Cornrnclinhomology between famjli(' ~ ha ... posed di nicult Phil yd ra t:cae) d irectly frolll a liliid l i rlt'a ~ ( '. ~l']JaI problc ilis without deta iled de\'doprllL'lI tal a nalysi" nectaries. ve... :-cl ... prirnaril~ in 11)(' root .... and "'('\ n al for ma n)' of the structures (Ki rdlofL 1991 ). c he mical ciwra(·ter:' (e.g ._ I"Ill·lidonit' iwid) fOlllld The Il Ulllcro us aU lapo rnor phi t· ...; u r th e Z ingi bc r ­ ill the I.ingilwralcs s u pport tllt 'ir pla t"t"IIH"ll t \\ ilh a ales ha ve a l ~o Iliade diffie nll lilt' dt'l nminalio ll o f lili a lcall l i n ca~(' (T a khlajan. 1980). Ihe evolu lionary relalio nship o f tilt' o rder 10 o lhe r T he li se of molccular c har 1...1\ t' gCllerally been yscs has !tn'n hig hl y Stu·c(' ...... ful in plall t... (1'':IIIIU'r allied wilh the Bromeliales and or COlllmelinalcs e t a l. . 1988: Crawford. 19(0). pa r t il"lilarl ~ illla \ ­ ( Il u ic hinson . 19i3: Dahlgrell ('I al .. 1985: T horne. ollolllica ll y diflinllt group" ill Idlich it i ... hard 10 1992). T he p re:o;.e ll ce of s tarc h y (·nduspe nn. cpi. intc rp ret morphological Ir o illolog~ (c./-! .. Cha ... t· & I"lltie ular wax or the Sln'lil:.ifl type. L· \'-fluores· Palmer. 1989: Smilh. 1991). H l' t'l'll ll ~ . ..; t·q lll·rw(' ("(! lIt o rganic M'ids in th(' ("{,II \\all--. and Iwo or fou r da ta derived frolll the- chloropia ... t gt'lH' ,.Iw l .. "hich :-i ub:,id ia r y cell::. in the S\Ollia lal ("oili plex a re a ll ('nc()( I (· ~ the large- s ubu nit o r ri bulose "i ~ l' ll u :-o pll . 197 k T akhlajan_ 1980: usc rul for taxonolllit: iln t'"ligatiun:-o al alld abo\"{' C ronquist. 19i5. 1981) based on the sim ilarit ), o f Ihe gell eri(' 1('\"t·1. 1I 0\\e\l'r. Ihl' li mi t:- ,IIHI taxo­ in fl orescencc a nd flowe r sl n H·turt·" (primaril y the nomic range ro r wh ic h rlw l . ca ll ... dl'q lla h" ly re"ol \, (' large. conspiclious hrac ls a lld lJt'laloid pe r ia n th ph ylog(·nc li t.· relationsh ip :-. have 1I 0t I wt'li t'xamilll'd paris ). All hough these mo r phologit:al homologies widely. arc pote n tially eq u ivocal. thc I'rt.':-. t' IlCC o f :'l e"cra l T o com pa re phylogcIli6 ha:"'d 0 11 Illllrl'hological c he m ical cons titllents (myr icctin alld or que rcetin a nal yses ill the Zi ll gi beralt·:-'. we init ialt'd a dadi"lic glyt'osid('s ) a l ~ o has s ugges ted a (' 01111 11 011 a ncestor a nalysis or rlJf' 1. scquCIll'C' da ta. O ur go.. li ..; ill thi,. fo r Ihe Z ingibe rales a nd Br o ll lt' l ia l~ ' s (Willia ms & researc h wert': ( I ) 10 exalllitlf' II II' 1I1011uph yl) of 622 Annals of the Missouri Botanical Garden

a n a nal ysis. Species. collection loca lit ies, a nd voucher information a re listed ill Table I . Total genomic DNA was extrac ted fr om fr esh or frozen leaf tissue by a modified CTAB method (Smith e t al.. 1991 [ 1992]). An approximately 1.40 I bp segment of double,sl randed DN A con·

10> 1 11> 1 SO"'" 11 > 1 l b l 10 >0 . b. taining the seque nce for the rbf·L genc was 3m­ " ,.0 ,,>I 2< >(1 50> 1 plified via the Polyme rase Chain Reaction (Cetus ,, ~ Corpora t ion) (PC B). 'I'wo s yn t he t ic oligOlluc leot ides we re used as a mplifi cati on primers. T he 5' prilne r It> 1 11> 1 IT» is the Z- I rbr L prime r based on the fir st 30 bp of 10> 1 ",. the rbc L sequence of maize, a nd the 3' prime r is I I the corresponding Z· 13 75 R prime r, whi ch is a 26 , ~• b , bp prime r deri ved from position 1375 - 140 1 of t.,. I ,,>. the maize sequence (Zurawski , DNA X). Initia l a t· l b • te mpts we re made to a mplify DNA from zi ngiber­ " .lb . alean ta xa using the primers of Olmstead e t al. J . > L ( 1992); howeve r, an apparent substitution unique to the Zingi be rales in this region of the gene pre· ve nted a mplification with these prime rs. This sub· stitution is c urrently under investi gatio n (Smit h el ", a l. . unpublished ). " , .,. Sequences we re obta ined by cloning the PCH pnx luct into BlucScript S K + (Stratagene, Inc.) us· ing e ither Ihe /li" e /I or 1..' ('o N I' sit c. The liga tion was fa c il ita ted by fir st in cuba ting Ih e PC R produc ts wi th DNA pol yme rase to assure blunt c nds . The products of the ligation reactions were used to Ira nsforlll compe tc nt cclls of 1:',~(' lll'ri('i(l ("0/ i (X L· I h" , Bl ues; Stra tagenc. Inc .). Ac ti vel y growing liquid i< ,. I cultures of transformed bac te ri a we re inoculated lb. " " with the helpe r phage VCS·M 13 (Stra tagene. Inc.) I and sin gle.stranded DNA wa s harvcsted that con· FIt": lI Ut: 2. Most parsimonious tree of the Zingibcralcs tained thc inse rted rbcL gcnc. This sin gle.stranded from Kress (1990, his figure 7). DNA wa s thell sequenced using Sequenase ve rsion 2.0 (US Bioche micals). a lld fragnIC nts we re se p­ a ra ted on 4% polya c rylamide gels. Interna l se· the eight recognized families; (2) 10 construc t the quencing prime rs we rc de rived from sequences dis­ phylogenetic relationships of the families wi thin the tributed by C . Zurawski (DNAX ). order based on rbcL scquc lIl:c data: alld (3) tu Addit ional sequences for !.ilium . .\J agrlOfio. de te rmine the siste r group relationship of the Zi ll ­ I 'ello:: ;o, P lll'O, Ti (fond sio. 81 {'gole pi ,~. P Oll /(,d e· giberules. ria. I.och"ollll!f's , Trat/,·sf"(lfl lia. a lld Moranlo we re graciously provid ed by colleagues (see Chasc MATE IU ALS AND MET HODS cl al.. 1993). S pecies we fe selc<.: lcd in a ll a ll empt 10 rc prcs('l1t Sequences we rc rcad directl y from the autora· the mosl di vergent Incml:W fs of eac h fa mi ly. For diographs alld e nt e red int o a NEX US fil e. This fil e example. in the large family Zingiberaceae. the was read inlo PA UP versioll 3 .0 s (Swofford . 1991) species selected represent each of th e four tribe::;. for cladistic a nalysis. Characters we re direc tly Din'erellt genera we re represented wherever pos­ scored for each lIucleotide and not mod ifi ed ill a ll Y sible. depending on availabilit y and number of gen­ wa y. i\ lissing data or a mbiguous regions we re scored era in each fa mi ly. A mi ni mum of two taxa per as mi ssing. Init ial a nalyses II st·d II EU BISTIC farnil y was chosen to redu(:e potcntial 10ll g bra nch SEA HCI-I and STEPWISE ADD IT ION or 500 c ffects (Fclscnsteill . 1978). whic h result whe n iJ IlANDOM ADD IT ION SEQUENCE ,epl;calc" sin gle taxon with no d ose a ffinit ies is in cluded ill TRB hrallch swapping. saving ALL M IN IMAL Volume 80. Number 3 Smith et al. 623 1993 Phylogenetic Analysis of Zingiberales

TBF:ES. and COLLA PSINC ZE BO I.I-:'1CT II the relationships o f .'-ilf';,:o(('pi,\ a nd the BrollH'li­ I~ B ANC II ES. Tlil'St' opliOlI ~ were l:hO~(' 1 1 as ighting a n a l y~.;(':-.. Neig hhorhood tret's of I alld oplion. }\I ollly four nodc ,..; did Ihc di .., trihuliorr ,.. 2 SIPps longer wen' al~o examincd IIsing the .";<1 111(.' differ in Inore thall two charact!'r :

Several separa te l:l ll aly!'('s \\(' f f' perfurrlH'd w.illg the c ha racter r-.la te di ~ tr ihll i io ll of til(' 1II 0,., t pa r ,.. i­ III(' a ho ve optiolls 10 df'\ennillc if c hoice of oulgroup mOllious tr ('e ~ . 709 ( ..... 61 (' ; ) of the I" t 6 : ~ ,'ha rach-r had all effect 0 11 the arrangclllcllt of ta xa \\ithill s la\(' c ha ng('s were trall ,., il i o n ~ . a nd \.1') 1 ( - :{9' ; ) the Zingibe r a l ('~. The ~e allaly~e s u ~ed: (I) JlaK­ wen' tr,-IIL s v(' r:;; ions Crahle :q, I/olia as the ollt~ro llp. wi th all other 1lI0ntwots The s trict cOlIsen,., u" o f th,' 62 treel' of 1.1 () I included a s ingroup: (2) I.i fill'" a :- Ill(' outgrollp ste ps or f(' \\('r l o"e ~ lIlo,.. t of the re'olutio lL foulld \Ii th all olher 1lJ0llo("ob included a :; illgrollp. a nd in the mos t par ~ illl o ilioll" tr,·e .. (Fig,.. . 3. I ), \ "" ,m 'h \/o/!flolia excluded: (:q '/'ffld(> .~(,(III/i(/ l'IIII/I·(Ir·. for trees two Slt'ps long"r thall the 1110 :-. 1 l'a r"IIIIO­ rill I .a("hn(ll////(,.~ a:-; oll tg rollp . with Old) tile zi n­ nious Ire{'~ produced t . S · ~ 1 In'es of 1.1 () ;) ,.. tl'l' ~ j.!;illl'ralean taxa included a.-; ingroup. or fewer. i\ :-Iri{'\ (·o nH·n .. u :-. of the ...:e I.S It In-t''''

Character s tate c ha nge,.. \1('1'(' plotted onto tree..,; lost ncarly all re;;; olulioll \\itll the ('\.('('ption of u ..: ing the ACCTHr\\ option. B('('all '-:(' the DEL­ monophyly of the fiY(· major clade .. in till' ZIII/!i­ TBA'\ option can !'olllC'tiJrH'''' dra .. ti('all~ alter the be r a les. St reli tziaccae. Zi np:il)(' ra('('a ('. e a r 1I1 'H'eac. di.-.;trihution whell ('( l1lall y p a r ~ illlOl lioll " opt ion:- arc Lowiac('ae. alltl (.,, /,,/1/1'(1 \/f/f'(/fI/oc fl lof/. a :-. IH'1I availahle. c hara(' tcr s tate changcs abo wen' plolted as OUl~rollP clades (Fig . I). with this oplioll. ami tll(, two distrihutions l'OIll­ IHlred.

A ciacii1- tic analy,.. i.. of IIII' "[)(' 1. :-eqll(' I[(," d a la p rodul:('d a :,ingle mo ~ t par,.. illlolliou" 11'(', ' for till' famili c;;; of Ihe ZingilH' I' :Ii ('''' (Fig. : ~ ) th"tI differ,.,

St'qucm.: cs of I .:~ · ~ S hI' or ilion: \\t'rt' ulJtailll:d from a ny pr{'vious pllylogt·llt·tit ' allo3l ~,.. i ,., of Illor­ phological dala (Figs. I. ~) (Krc "." . 1990). Thl' fo r :U s pecies including I 0 ~(.'q u ('llt · t · :- gnwiollsly Lowiaccae a nd SlreliI7iact ·.w lincage i-- till' (J111 ~ provided by other \\orkC'r:- (:- e(' Ckl:-" I't al.. t 99:~) between-famil\' d ade !-ohared 1)\ hoth the IIIOll'l 'utar (Taillt.· I ), For tltt· : ~2 :- pe('ie,.. u:-ed ill the majority . - a nal ysis and Oll{, o f Ih£' ('Iad i... lic alla lv:-.(· ", of 11101' - o f a nalyses (exeludill~ \/a/!lIo/ia. Idlidl \\<\.-.; lI ~ e d - . 10 d etermine olltgrollp rclation...: II ip,.,). 16·1 I'0,.,ilioll:­ phologit':tI charactcrs ( Da h lgren 8." Ib ... IIIU .... t·lI. I 985)(Fig. I ). (-: ~ 1C; ) were va ria hit.· , or til(':-(' 16,I charaders. The ('onfli"ls bet\\{'('1L tilt' tn'(·,., Ila--{'d Oil mor· 18:~ (-39C-:- of Ilw va ria hle characters ) \\I'r(' au­ ta polllorphic . Therefore. 28 1 of the original 1.:i,J, 5 phological ,·haraclers alld molt'('ular d .J ta a n ' many. C One conspicuous cxample I't'rtain ~ to flora l mor­ lip (-2I { ) were ,..; hared Il y 1\\ 0 or more taxa, Of phology. In a ll traditional (' la.-.; :-; ifi l'ation:-. oftlH' ord"r Ihe · ~64 variable char a Cl1-r~. lOS (-2:~ (; ) \\'c re 1 (see Kr e:-; ~ . 1990. for :- lltl1llL '- lr ~) . the n 'dllt'lion ill fir ... 1 codon PO!'ilion...:. 92 ( -- 20 '( ) \\CIT ,..t:('olld po­ l'itioll"', a nd 267 (- 57('; ) \\erc third I'o;-, itioll'" (Ta. the number of pollen-hearillg ~ talllell :- fro m h or :) hit- 2). ( ~ l u sMaranta('('ue). ami the a !-o,.,o,·ial,·d ,\Ni\t.Y:-;ES modificalion of IhcsC' slalllt·II .-; illto petaluid ,.. Ialll;­ The cqually wei~ ht('d anal~,.,il' from thc HA N­ nodes. af(' (' h a r a (' t e r ~ that Il a\T ""ened a ,.. tll(' Ila , i... DOli ADDITIO, ~F.QL · I ·: :,\CI-: , carel. rc, uit ed ;n for dividing Ihc famili£', intn 1\10 ha ~ i (' g roll p ... . the t\\O 1II0 ~ t par ~ i mo ll iou;;; trce:, of 1.16.1 ,.. teps «(: 1 of ba na na grou p a nd Ih(' /! inp: t·r group. re" pCl'tl\ , · I ~. O, · ~I; HI of O. · ~9). T lw t wo trees differed o nl )' ill The homology of Ihest· "' I)('cialized flor"l ft ·"Iun·:- 624 Annals of the Missouri Botanical Garden

TABLE 1. Sources of rbel sequences (all material is deposited at US. SEL. or DUKE). Voucher and Genbonk information for sequences obtained from other la boratories is referenced in the appendix to this volume.

Species Voucher Source Cenbank #

Dromeliaceae Tilluflc/sia eii:lIbetlw(' D. Clark PU )'l1 dyckioides D. Clark Rnpateaceae S t cgo l ep i .~ allcllii D. Clark Commelinaceae

Traliescafllia SOCOfWS('(/lU/ ~:I a l udu Faden 76·98 Mexico L05463 'f'radescunlia sp. M. Duvall POll tederiaceae POfl tederitl D. Clark Haemodoraceae Lach !l(wt hes M. Chase Vel loziu ceae Vrllo:ia D. Clark Litia ceae I.i/ium superbllt1l M. Chase Typhaceae Ty pha {alifolia L Kress 90·3 170 Maryland, USA L05464 Typha lallfo/ia L. M. Duvall Cannaccae Canfla jndica L. Kress 80- t 124 Indonesia L05445 Cafllla lu erkheimi; Krauzlin Kress 76·653 Panama L05446 Costaceae COSIIIS barbatlls Suess. SEL 86·0550 Marie Sclby L05447 Bota nical Gardens, Sarasota. Florida Tapei flochilO$ (lflaf/(l s .~(I(' K. Schum. Kress 79· 1114 Duke Uni"ersit y, L05462 Durham. North Caroli na M OflOCOM IlS !HI/florus (Pocpp. ex Kress 79- 1112 Peru Petersen) Maas L05454 Heliconiaceae /-Ielicoflia la tispathll Bcnth. SEL 80·16 10 Marie Selby L0545 1 Botanical Gardens. Sarasota. Florida /-Ielicol/ia pah A. C. Smith Kress 79· \ 072 Fiji L05452 Lowia ceae OT('hit/afltha fimbria/a Holtlllm Kress & Beach Malaysia L05456 87·2 159 Orchit/afltha .~ illf1H''' si$ K. Larsen Kress 92·3468 Malaysia L05457 Marantaceae 111(lr(ll/t(l /(,IIl' (!/U'urli M. Dll\'a ll C(l/(lIII('(I 1t)('w'lI('ri 1\ lacIJridc SE L 85·31 US Botanic Garden, L05444 Washington. D.C. Marrllltoch/o(l p'lrpllr('(1 (Ridley) Kress 78 -894 Wilson Botanical L05453 Mil ne- Redhead Gardcll. ColOtn Rica Mllsa ccae EI/ .~I' /I' 1'I'II/riCOSIIIII (\Vdw.) s.n. Wilson I30 tanical L05448 Cheesman Garden. Costa n ica Volume 80, Number 3 Smith et at. 625 1993 Phylogenetic Analysis of Zingiberales

T,.' BI.E I. Continued.

Species Voucher Source Gellbank 1+

J/II .HI " (' /III/ ina/a Colla !>. n. US Bolanic Garden. 1.05455 Washington, D.C.

Strelitziaccac

PhpI/(1 ko .~ prrmll m gllia fle" s; s Kret-t- 86·2099 French Guiana 1.05·158 (L Riehl.) Miq. R (IN' lIfl la /1/a dagasc(l fi(' flsis K rc~~ 92·350 · ~ US Bot anic Garden, 1.05·159 1. F. Grne!. Wa"hinglon, D,C. Srreiir: ia flicola; Regel & Koch Krc",s 91·3 169 US Botanic Garden, 1.05 461 Washington. D.C. Zingibcraccac G/fI/,I1fI l'llr/isii I-I Olt tUlll Kr i '~ S &: Beach 1.(154·19 I3 j.2 16 1 l lS IlG 90·65:1 US Uotanic Garden. W a~hin~ton. D.C. SEL 83·203 Lyoll Arboretllin. 1.0.')4-60 Iionolulu. Hawa ii 7.ifll!.ilU'r p.(([lfIilU'lllfI Nororllm Kn.' t-~ 91·3266 L vol ! Arboretum . 1.0.')..\- 6S • Honolulu. Hawaii

i:-- s upported by ontogenetic s tudie:'> in the Zi ngi­ exalninatio n o f o lhe r molec ula r (' h a r aclt'r ~) IJefo re !'e ra les 1.1:-; weI! ( Kirc hoff. 1983. 1988. 199 1), Al­ concluding Iha l the mo rphological fca tll rl's arC' ho- though the four families o f the ha na lla g roup rna y 1l10piasl ic. be \'ar i o \l ~ l y rela ted . the ginger grollI'. de fill ed by T he rapid "d('('ay" of t he va rious ("!;Hh· ...; ill lhe th e~c hig hl y derived s taminal featu re". has a lwa ys mos t pars imoniou ,., tree found from Il l(' r/J(' 1. dala been eo n ~ idered mo nophyle tic ( Dahlg rcn & Has­ stlggests that III(' illterfamilial phylog('lwtic ~ig nal trlUssen. 1985: Kress. 1990; K irchoff. 1991 ). of Ihis plas tid W'lLe is low for the Zi ll ~iI)t'ral(' !'> (Fig, T he to pology ha sed on the Illol('("ubr d a ta places 4). The re are 62 d iffe rent tOJ> ologie !'> ,h,t! art' Olll.' the four families o f the ginger group inlo three ste p or fewer 10 llger than the mo:-; t i'ar !'> irnoniou:-;; :;e pn ra te IillCages ( Ma ra llt acC' ta nlCIIS is pie­ o f most of the families. Althoug h til(' I'o!'> ition of :; iolllorphi(') ill :-;t ('oll"lp:-c !'> in tllese t l' rohu !'> 1. III a criti(,a ll y (throug h irlC rea.'ied .'i ali l plill ~ of laxa, o r la rger a n a l ysi~ of the phylogeny of tht' IIIOliocols

T "I II~, 2, \ 'ariablc characters a<:cording to codon POSIIIOI!. Homoplasti c character !'t atcs a re b ,, ~ ~'(l 011 the ACCTRAN cha racter state distribution.. for the mosl parsimolliolls trees ( F i~ . 3), The fi rst ""Illes are tht' lIumber!' of character ~. the second are the pt'rcclllages of lolal states. Synapo!llorphil' character siaies includt' only non· homoplasti c character stales.

Posilion \' a riable Homoplastic SynapOlllorphic A ulaporrrorphic

First 105 - 22% 30 15% 23 - 28 0/( S2 _ 28 (1{ - _ 23 (T{ _ "'1(1' Second 92 - 20 (1 ~ 30 - 15% 19 · ~3 -' , Third -?6-, - SSC'; 139 - 70 'X 40 - 49f1 88 - , ~9 (' ( 1'01 .11 ·164 I 00(':' 199 100% 82 100('; 183 100"; 626 Annals of the Missouri Botanical Garden

46(31) Lilium 20(15) Costus 16(14 ) 15(3) I * 40(28) T a peinochilos co I 24(1) Monocostus (6) 25(4) Ca lathea 18(3) * ( 0) (6) Ma rantochloa MA 11 (0) Maranta 29( 8) Hedychium 10(8) 29( 8) Zingiber 11(0) 21(16) * Zl * 32(21) Globba 16(9) 18(0) (6) Riedelea * 20(10) HeHconia pa ka (4) HE 29(20) Heli coni a latispath a (3) 23( 0) Ravenala 12(8) 37(25) Strelitzia 9(7) ST 6(5) 51(30) Phenakosperm um (3) 29(21) 7(5) * 12(7) Orchidantha fimbri ata * 29(6) LO * Orchidantha siamensis 32(20) Ensete - MU (9) 15(0) Ca nna indica (2) CA * 5(4) 24(1) Ca nna tuerkheimii 15(1) Musa - MU 15(7) 15 8) Lachnanthes 23(2) 37(27) Pontederia 19(13) 28(8) Tradescantia soconuscana 48(3 1) 12(4) Tradescantia 6(4) Tillandsia 21(16) 1()(7) Puya 21(16) 13(7) 15(2) Typha 5(4) Typha 22(2) Slegolepis 44(31) Vellozia

FI(;[IIIE 3. St rict consensus of the two 111051 parsimonious trees from the equally weighted analysis. The Iwo trees differ ollly in the relationships of S/('go/f'pis T),pIUl / Brolilciiaccae. Numbers along branches indica te substitutions !lupponing thaI clade. numbers in l'arclllheses are the portion of substitutions thaI are homoplastic. Characler slat e change distributions are based on tree I of the IwO 111051 parsimonious trees (Ty pha as sister group to Bromcliaceae). and tht: ACCTRAN option. Asterisks denote clades ,hal are losl in the conscnslis of the 62 trees one step longer than the most parsimonious trees (see Fig. 4). Families of the Zingiberales are denoted as follows: CO = Costaceae. ~ I A = Marantaceae. ZI = Zingiberaceac, HE = I-I cliconiaceae. ST = Strditziaceac. LO = Lowiaceae. MU - Musaceae. CA = Carmaceae. Volume 80, Number 3 Smith et al. 627 1993 Phylogenetic Analysis of Zingiberales

C 06tUI T lpo:-inochiJ08 M0l'lOCD51us C.\athl!l M"<1ntochloa M ... "ta

l-looychium Zingibo.. G'''''''' R~eIN Hell.,.," ... pi"" Ilclioon" 'ali.path. RavMarantochloa Can"" luc,kh<.'i mii

1-I...Jychium M'N Zlngiber I c,_ B Riedelea Helicon;. paka -{ Heliconi. Iatispatluo Rov.,",t, St,ditzilo Ph enl kos J>ft"m u m

Ordlid.nth fimb ri.ti Co6luS Ordiidantha .i.mensi. rlpeinochilO5 EnSoCle MonO<'06lU5 Corona india C .la,hea Cann. tuerkheimii Marlntochlm A M,~ Mar,"1a Hedychiurn I Zingibet Globb. RieddCII 11"lieon" paka Heliconi. lalispalt.. R..i v,",u.l. I SI,elitzi. I l'heruk06permum

Orchid. nth. fim b r i,t~ Orchid.llth.. 51' mcn!;l5 EllSol'le unn. indica Canna lue. lche,mi, MuSIl c

FICIlIIE 4 . Strict consensus trees of the topologies based on A: two trec ~ of J, 163 "tep)'; each, 11: 62 tree" of I. 16--l steps each, and C: J ,541 trec~ of 1,165 steps each. Only taxa of lilt' Zingiberalcs arc illu;;tratcd ill thc~t' fi gures.

La sed on rhd _ sequc rH'C data (Du vall c t al., 1 99:~ ). T he Musa ce,w (I'araphylt'lic wilh 1he Call 1la ­ the sis te r group relatio ns hips or tht' Zingibc ralc s ccae) arc the onl y ralll il y ill lilt' /ill gilJcra l, ';; that a nd the Ilaclllodor a{'cac Ponll'deriaceae CO III ­ is nOl monophyl"lic ill til{' lIIo !'> 1 par!'> illlolliou:-- tn't'. lIIelinaceae is !i upportt'li. The monophyly of th e IId iI'O lli

TABLE 3. Type of character state changes based Oil sequence of the resulting protein. Howeve r, the tree I of the most parsimoniolls trees (Typha ami Bro­ hig h pe rcentage of homopla stic third position co­ IIlciiaceae liS sister groups) in the c(lua ll y weighted ana l }'s i ~ dOllS (139 of the 464 variable positions) .md the (Fig. 3 ). Transvcrsiolls arc imiica lcJ ill bold. prese nce of 3 - 4 diffe rent lIucleotides at 64 of these sites indica te that the substitution rate wi thin the A c G T Zingibe ra les Illa y be close to sa turation. thereby A - t03 27 6 110 reduc in g the int erpre table phylogene tic signal of C 137 43 3 the ,bel sequence da ta . G - 104 Our results suggest tha t although phylogene tic signal is present in the data. the re is a "window" in wh ic h ,bel seq uence da ta docs not strongly the consensus of trees O IlC step longer. These dis­ resolve phylogene tic rela tionships. For the Zin gi­ c re panc ies may be the result of li mited sa mpling. be rales this window is a t the between-falllily leve l. Sequences for addit ionai llelico"ia and Carlf/a spe­ At the ordinal and famil y le vels, the molecular data cies, as well as other represent ati ves of the Mu ­ a re much more robust in defining and resolving saccae. may stabilize these port ions of the trec. ph ylogenet ic rei a t ionships. The other six famil ies are we ll support ed as mono­ An explanation for this window may be related ph yle tic groups by the molecul a r data even in the to the age of the Zingibe rales and the tilLle since less parsimonious trees. di ve rgence of the fCretaceous. In cOntrast, ph yloge ne tic res­ more robust a nalysis and provide a better ove rall olut ion of lineages within the Asteridae sensu lato estima te of phylogeny for the Zillgiberalcs . In par­ is resolved adequa tel y with rbe l seque nce data ti c ular.the monophyly of these fa llLili es would prob­ (Olmstead ej al. . 1992). a bl y be more strongly supported by the addit io n Extant Zingibe rales possess numerous deri ved of taxa to the a nalysis. Illorphological features, a nd five of the e ight fam ­ Hegardless of the proble m of uneven ta xon sam­ il ies are known frolllthe fossil record (Kress. 1990). pl ing. the limita tions of the data se t are 1I10st a p­ Although most of the fossil ma te rial has been col­ pa rent whe n the distribution of charac ter sta te lec ted in Eocene deposit s, the old est spec ime ns are changes a re mapped onto the tree (Fig _ 3). Of the leaves of the Zingibe raceae from the la te Creta­ 28 1 phylogenetica ll y illfonnativc c haracte rs used ceous (Hi cke y & Peterso n. 1978). The common in the a nal ysis. the re a rc o lLl y 90 (32% ) synapo­ ancestor of the lineages leading to the banana group tlLorphic c ha racter state changes that a re not ho­ is the re fore hypothesized to ha ve di verged from moplasti c (based on Fig. 3). \Vithin the Zingibe r­ the re mainder of the Zi ngi berales by the la te Cre­ a les. the re is onl y one charac ter ~ t a t e c hange tha t taceous (Friedric h. 1987). suggesting that the lila ­ is synapomorphic betwecn fa milies and not ho­ jor lin eages within the order had rapidly diffe re n­ moplastic (Strelit zia ceae a nd Lowiaceae lineage). tiated by the earl y T e rtiary. These times of lineage In co ntrast. seven nonhollloplastic cha rac te r state splilling and di verge nce in the Zingibe ra les are c ha nges support the monorhyly of the orde r. a nd similar to those descri bed by Olmstead e t al. ( 1992) 29 non homoplastic characler state changes sup­ for the higher di cots. The la ck of ph ylogenetic port monorhyly of, or arc syna pomorphic wi thilL . resolution usi ng ,brl data in these unre lat ed fl ow . the families of the Zillgibe rales. likewise. the e ring plant taxa ma y be due to their common age mo nophyly of the olL tgroup clades (Haemodora ­ of origin and di ve rsifi ca tion. c eae/ Po ntede riaceae/ Comme lilla ceae a ll d Bro­ To further cla rify ph ylogenetic relationships meiiaceae/ T yphaceae/ Ra l)ateaceae) is supported within the Zingibera les, data frolll other sources by nonhomoplastic c harac tc r states (Fig. 3). will be necessa ry. The morphological da ta wi ll nec ­ The wea kness of the sig na l is a lso a ppare nt whe n essa rily be re-examined to accollJmodate new in­ codon posit ion is examined . Third positi on codons te rpretations of la o,nology a nd to include taxa such account for 58 % of the va riation in the data se t tha t the molecular and morphological da ta se ts will a nd 70% of the hmnoplasy Cr a ble 2). This is not he direc tl y cOllll-'a rable. Seque nce data from the a surprisin g result as the redundanc y of the gene tic nuclear e ncoded 18S a nd 26S ribosoma l genes are code permits a hi gher rat e of substitutio n a t third currently being collec ted for the Zingiberales (Kress position codons without ~ Ilt e rin g the amino ac id et a I. , unpublished results). which have bee n suc- Volume 80, Number 3 Smith et al. 629 1993 Phylogenetic Analysis of Zingiberales

ccssful in resoking some ancient ph ylogene tic events FIl ]EDJlICIl , W. 1987. The evolulion of the Zingiberab in other seed pla nts (Ha mby & Zimlner. 1992). during the Cretaceous and Tertiary. XI\' Interna­ Additional data may a lso be obt a ill ed by restricti on tional Botanical Congress . Berlin. Abstract rtulllbt' r 5-30-6,284_ sit e compa rison of the highly conserved inverted HAMil)'. R. K. & E. A. ZI\I\I EH. 1992. RibosOIlial RNA repeat regioll s of the chloroplast genome (Down ie as a Phylogenetic Tool in Plant Systematics. Pp. 50 &r Palmer. 1992: Sill ith et aL ullpubli:;hcd r<:.<; ul1 ,,) . 91 ill P. S. Soltis. D. E. Soltis & J. J. Doyle (editor!-o), Molecular Systeltlatics of Plants. Chapman & lI alL New York. LrTEnATl'llE C ITED HI CKEY. L J. & R. K. P ETEIN ):".. 1978. 7.ill,:iuerolms, BE:\TII,\.\I, C. & J. D. HOOK EH. 1883. Genera Plantar· a fossil gcnus of the ginger fami ly frotTI La te Crc· lIlll, \'01. 3. L. Reeve & Co .. 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