Generic boundaries and evolution of characters in the Arctium group

Generic boundaries and evolution of characters in
the Arctium group: a nuclear and chloroplast
DNA analysis
A.
SUSANNA,
N.
GARCIA-JACAS,
R.
VILATERSANA
& T.
GARNATJE
RESUMEN
SUSANNA, A., N. GARCIA-JACAS. R. VILATERSANA
& T. GARNATJE (2003). Gencric boundaries
and evolUlion of characlers in (he Arclilllll group: a nuclear and chloroplasl DNA analysis.
Callee'. BOl. (Bllrce!OIU1) 26: 101-118.
Definir los límites genéricos en el grupo Arc/;/l1/I (Composi/(ll!. Car(/ul!al!-C{/l'dllilwe).
formado por los géneros ArctiulI1, COlIsillitl. Hypac(IIl/!liulI1 y Schlllal1wlIsell;a, ha resultado ser
una tarea muy complicada. Concretamente, la separación precisu de Are/il/m y COl/sillia es
muy difrcil de establecer. En consecuencia. hemos :lOaliz.ado las secuencias del DNA de dos
regiones. el gen cloropltistico marK y los espaciadores ITS 1 y 2 del ONA ribos6mico-nuc1cllT.
de una amplia representación de todos los géneros del grupo: en el caso de CO/l.~il1ia,
centrándonos en las especies más obviamente relacionadas con Arctilflll, ¡'Iemos cncontrtldo
una correlaci6n exacta entre Iilogenia molecular y dos caracteres fundamentales. el tipo de
polen y el número cromos6mico: todas las especies estudiadas que lienen el tipo de polen
Arctiastmll1 y el número x= 18, característicos de Arctilllll sensu stricto. forman un ciado
monofilético. hermano de ouo ciado monofilético formado por las especies de COlIsillia sensu
suicto. Sin embargo. el ciado "Arclioide" no se puede definir con caracteres morfol6gicos
macrosc6picos: el carácter que separa Arctil/lI1 y COIlSillia son las hojas espinoso·pinnatifidas
o pinnatisectas de COlIsiltia, que es adaptativo y tiene poca relevancia taxonómica. Según
nuestros resultados, las espinas habrían aparecido en dos linajes diferelllcs: en COl/sillia. por
una parte, y en H)'fJacanr/¡irllll y Sdll/rtlllral/sel1ia. que son espinosos y morfológicamente más
pr6ximos a COlIsinia, pero que están sin duda más relacionados con Arctil/lIl, inerme.
Evaluamos también las implicaciones de esta incongruencia entre morfologfa, por un lado, y
datos moleculares, palinol6gicos y cariológicos, por otro. Proponemos algunas soluciones, pero
ninguna es totalmeme satisfactoria: se necesitan más estudios que incluyan más especies de
COl/sillia subg. H)'fJacanrltotles.
Palabras clave: Compositae, Arclilllll, COl/sil/io, Hypaclllllhilllll, ScllIIrtllI/(lI/sCl1itl. cvolución,
Iilogenia molecular.
Abstraet
SUSANNA, A., N, OARCIA-JACAS, R. VILATERSANA & T. GARNATJE (2003), Generic boundaries
and evolution of chllracters in lhe Arc/il/III group: a nuclear and chloroplast DNA analysis,
Collec/, BOl. (BlIrcelolw) 26: 101-118.
Oeneric delineation within the Arctil/lI1 group (Colllposi/(/c, Ct/n/ueac-Caf(/l/il1ac), formed
by (he genertl Arctilll/l. COl/sin;a, HYI'{/cmrlhilllll and Sdrllllllhol/senia, has proven a cornplicated task. In panicular. the precise limits betwccn Arc/il/III and COlIsillia are vcry difficuh to
establish. Therefore, we have carried out a molecular survey of DNA sequences of two regions.
the chloroplast gene II1l1tK and the lluclear-ribosornal spacers ITS I and 2, of a represe mm ion
of alllhe genera of the group (in the case of COlIsillia, centred in the species more obviously
related to Arc/irllll). Our results show a precise eorrellllion belwecn molecular phylogcny and
Alfonso Susanna, Núria Gareia-Jacas, Roser Vilatersana & Teresa Garnalje. BOlanicallnslilulC of
Barcelona (C.S.I.C.-Ajuntamenl de Barcelona), Pg. del Migdia. s/n .• E-08038 Barcelona (Spain).
AUlhor for correspondence: A. Susanna (asusanna@ibb.csic.es)
102
COLLECTANEA BOTANICA (BARCELONA) 26. 2003
two very important characters. pollen type and chromosome numbers: all Ihe investignted
species wilh lhe ArctiaSlrllI11 pollen Iype and x= 18. characlerislics of Arrt;lIm sensu 5lneto.
forro a monophylclic c1ade. sister lO another monophylctic dade Cormed by allthe invesligalcd
species of COllsinill sensu slrictO. However. (he resulting ..Arc:tioid" c1ade canno! be defined on
macroscopic morphologic charnclers. because Ihe mnin Il1Iil for segregaling Arctilllll and
COI/$in;a. (he spiny pinn:l.lifid-pinnatisecl ¡caves of COIIsinia. is adllplnlive and of scaree
syslemalic relevaoce. In facl. OUT resulls suggeSI Ihll( spines have appeared al Icnsl in two
differenl lineages: (he genera
HypaClllII/¡ill/ll
and Schmalllallsenia. spiny and thus morphologi·
cally doser lO COl/si/lia. are unambiguously relllted to Ihe unarmed genus Arelil/III. A hypoth.
esis on lhe evolution of morphology, pollen and chromosome numbers in the group is
formul:ned. The systematic implications of this incongruence between molecular. pollen and
karyology. on the onc hand. and morphology, on the other h:md, are evaluated. Sorne possible
solutions are proposed. bul none of Ihem is tOlally satisfactory: more studies nre necessary wilh
Ihe inc1usion of ncw speeies of COl/sinia subgenus Hypaclll1/l1odes.
Key words: Composi((le, ArClil/lIl. COl/sillia, HYfJacal1/hil/lI1. Sc/¡malJrfIllSeI1Ü,. evolution,
molcculnr phylogeny.
[NTRODUCTION
The tribe Cardueae has always been a conflicting group, and both tribal and subtribal
delineaIion has been much disputed (CASSINI, 1819; BENTHAM, 1873; HOFFMANN. 1894;
WAGENITZ, 1976; OIITRICH, 1977; PETIT el al., 1996; PETIT, 1997; BREMER, 1994). However, all surveys on Ihe basis of ONA analyses support Ihe monophyly of Ihe lribe: cpONA
reslriclion sile dala (JANSEN el al., 1990, 1991), cpONA rbcL gene (KIM el al., 1992),
cpONA marK gene (GARCIA-JACAS el al., 2(02) and nrONA spacers ITS 1 and 2 (SUSANNA el
al., 1995; GARCIA-JACAS el al., 2(02). The subtriballimits are more complicaled lo establish,
but the tradilional e1assification in four subtribes (Carlinillae, Ecllillopsidillae, Carduillae
and Cemal/reillae) remains the only practical approaeh (SUSANNA & GARCIA-JACAS, in
press) and is generally accepted.
Having acknowledged the tribal and subtribal limits, the next step was lo e1arify the
delimitation of sorne of the major genera in the tri be. Now, among the most interesting and
vexing problems of genus rank that persist in the Cardueae is the marking of lhe generic
boundaries in the Arelillm group.
The Arclilllll group
The Arclium group eomprises the genera Are/il/m L., Cousit¡ia Cass.. Hypacamllium Juz.
and Scllmallrallsenia C, Winkl. (H.i,FFNER, 2000; SUSANNA & GARCIA-JACAS, in press).
HAFFNER (2000) adds (he genera Upskydla Juz. and TIarocarplls Rech. f., merged into
COllsinia by SUSANNA & GARCIA-JACAS (in press).
Three important eharacters never found in combination elsewhere in the tribe characterize basically theAre/illm group. First, the receptaele has strongly twisted scales. Second, the
achenes are always tigrine (with a panero of wavy fringes), very often winged, and without
a neclary. Third, the pappus is formed by free deciduous bristles (SUSANNA & GARCIAJACAS, 2003). 80th on morphological and molecular grounds, the genera Arclium and
COIIsit¡ill belong to the subtribe Carduill11e. More precisely, the Aretium group is part of a
large e1ade ineluding also the subtribe Celllaureillae and the genera Saussl/rea Oc. and
JI/rillea Cass. (GARCIA-JACAS el aL, 2002).
Are/il/m eomprises 11 species, according to the latest c1assification of the genus
(DUISTERMAAT, 1996), most of lhem wilh a subcosmopolitan distribution. DUISTERMAAT
A. SUSANNA,
N.
GARCIA-JACAS, R. VILATERSANA
& T. GARNATJE
103
(1996) recognized four sections: Arcti1l11l sect. Arctilllll (Wilh all lhe species classically
included in Arctium), and three more sections described as sections of the genus Co//si"ia
by TSCHERNEVA (I988b, 1988c): sections Lappaceu11l, Nllllarctilll1l and Pselldarctiu/1/.
The large genus COLlsinia is formed of ca. 600 species (MAIlIlERLEY, 1990) located in lhe
Iranian and Turkeslanian mountain regions, with an aSlonishing number of ende mies. Both
lhe limils and lhe sectional classification of COllsi/lia have widely changed. Proposals by
BUNGE (1865), BOIssIER (1875), KUNTZE (1891), WINKLER (1892) and BORNMÜLLER (1916)
were superseded by TSCHERNEVA (1962) in her trealmenl for lhe Flora of lhe USSR. La<er
on, lhis treatmenl was again deeply modified with the proposal of two new subgenera and
many new sections and series (TSCHERNEVA, 1988a). The latest revision classified COllsillia
in lhree subgenera and 50 seelions (TSCHERNEVA, 1988b, 1988e).
The rest of lhe genera of the ArClill1ll group recognized by SUSANNA &GARCIA·1ACAS (in
press) are very small: Sc!lmal/¡ausellia is monotypic, and Hypacallt/¡illlll has only three
species. 80th genera are narrow endemics of the mounlains of CenLral Asia (Tien-Shan and
Pamir).
The eXlremely conflicling relalionships belween the genera of lhe Arcti/lf11 group are
reflected in lhe many reclassifications suggested in Lhe group thal imply changes of genus
adscriplion. We shall shortly commenL on sorne especially il1uslrative examples.
The prolologue of lhe genus Hypacallfhilllll placed it near SC/¡lIIal/¡ausellia. Previously,
it had been described as COllsinia ec/¡il1opifolia Bomrn. and combined as SC/¡lIIalhallsellia
ec/¡illopifolia (Bornm.) Juz. According Lo TSCHERNEVA (1983), Hypacllllr/¡iu/1/ was also
connected to Cousinia subgenus Hypacallt/¡odes Tschemeva sect. Lacerae, and related to
genus Sc/¡malltausenia.
In its lurn, Scfmllllllll/lSellia was firsL described as COllsillia eriophora Regel & Schmalh.
As we have seen aboye, it was al so relaled lo Hypacall1/¡illlll, and il was even combined as
Arctiu11l eriopltorum (Regel & Schmalh.) Kuntze.
These entangling cases are illustralive enough, but Lhe besL example 01' lhe problems of
generic range in the group is the case of Arcri///1/ and COllsi"ia.
Relationships between the genera Arclium and COllsillia
A c10se conneclion between both genera has been signalled fram old (B01SS1ER. 1875:
KUNTZE, 1891). AH recenl studies have confirmed lhis conneclion: morphological surveys
by DITIRICH, 1977; DUISTERMAAT, 1996, 1997; PETIT el al., 1996; PETIT, 1997; HAFFNER,
2000; or molecular analy,es by HAcFNER & HELLWIG, 1999; GARCIA-JACAS el al., 2002, are all
coincident.
Species of Arctiu11l and Cousillia have moved belween genera very often: Boissier (1875)
combined Lappa (=Arctill1l1 !) a"'plissima Boiss. as COlIsi/lia alllplissillla; and KUNTZE
(1891) proposed lhe c1assification of alJ the species of Co//si/lia in lhe genus ArctiulII. in
view of the impossibility of establishing the precise limils between botll genera.
The main obstacle for Lhe precise delineation between Arcti//I/I and Cousinia is the
presence of a group of COllsillia species (the "Arctioid" group for DUISTERMAAT, 1996) wilh
many characters shared with Arc,illm: above al1, lhe presence of glochidiale spines in (he
appendages of lhe bracts and the big unarmcd lea ves. The most recent proposa! was Ihe
Solulion suggested by DUISTERMAAT (1996) and followed by SUSANNA & GARCIA-JACAS (in
press): Lo c1assify lhe more obviously "Arclioid" Cousi/lia species (5 species from differenl
sections) into Arctiu11l, while exporling TSCHERNEVA 's (1988 b, 1988 c) secLionai c1assification of Lhese species of COlIsillia lo ArctiulIl. Soon after, DUISTERMAAT (1997) suggesled a
rectification of her own views, and poinled out vaguely Ihat all Lhe species of COllsi/lia subg.
Cynaroides Tscherneva could be placed in ArctiuIIl.
COLLECTANEA BOTANICA (BARCELONA) 26. 2003
104
We shall shortly explain lhe maio traits thal have been used in the delimitation of the
genera of the Are/j//m group: morphology, karyoiogy, pollen and geography:
Morphological characters: leaves, bracls and heads
Arcril/m leaves are always unanned, often very big (lO 80 cm) and usally cardate, bOlh
characlers being shared by COl/sinia subg. Cy"aroides and Hypacanthodes. In contrast, COl/sinia
subg. COl/sinia leaves are small, usually lanceolate and with few exceptions very spiny.
Involucral bracts are a key character. In Arcriwll sensu stricto and part of Cousillia
subgenus Cynaroides (the "Arctioid" species of COlls;lIia). involucral bracls always end in
a recurved hook. Instead, in most of (he species of Cousillia, involucral bracts end in a spine
withoul hook.
OUler florets in ArctiulIl often have longer and brighter coloured anlher-tubes, mimicking
ouler radianl ligules. Instead, many COIlsillia species often have bright-coloured appendages in the innermost bracls, recalling those of Carlilla L.
As lO floral morphology, DUISTERMAAT (1996) concluded lhal the stigma of COllsinia
subg. COl/sil/ia was very different from lhe stigma of Arctium and the "Arctioid" species.
For DUl5TERMAAT (1996), COl/sillia subg. COllsinia lacked the hairy lhickening below the
branehes lhal is a key eharaeler for lhe lribe Cardueae. HiiFFNER (2000) noled lhal lhe
sweeping ring of hairs was also absent in Schmalhallse"ia.
Habil
According to DUISTERMAAT (1996), rnonocarpic habil is a key characler for Arelilllll.
Instead, COllsinia species are, with only a few exceptions, perennials. Sorne "Arctioid"
species of Co//sinia are rnonocarpic, bUl rnost of them are perennials (TSCHERNEVA, 1962).
Dispersal is also differenl. 80th genera share a [eature infrequent in lhe tri be: achenes are
dispersed logelher with the heads. In Arctillm and most of the "Arctioid" group, propagules
are usually the whole heads, dispersed by exozoochory by means of the hooks of the
involucral bracts. On the contrary, mosl species of COllsillia are turnble weeds, like rnany
plants fram the steppe: the whole plant is dispersed by the wind.
Chromosorne nurnbers
According to literature (revised in DUI5TERMAAT, 1996), ArcliulII always has x;;;; 18 and
211 ;;;; 36. The same number is shared by aH the studied species of COllsillia from Ihe
subgenera CYlloroides and Hypacalltliodes (TSCHERNEVA, 1985). The genus Sehmolho//sellio
also has 211 = 36 (SUSANNA el al., 2003). This high number, lhe highesl in all lhe Cardlleae,
suggesl that ArClil/m, CO/lsinio subg. CYllaroides and HypocomllOdes, and Schmalhallsellia,
eonslilUle old polyploid eomplexes (TSCHERNEVA, 1988 e).
COl/sil/io subgenus COl/sinia has 211;;;; 18?, 20? [doubtful to usl, 22, 24 and 26
(MOORE, 1973. ¡ 977; GOLDBLATT, 1981, 1988; GOLDBLATT & 10HNSON, 1990, 1991;
GHAFFARI & DJAVADI, 1998; GHAFFAR] el al., 2000; SUSANNA el al., 2003). This long and
complex disploid series is a proof that both groups have evolved in differenl ways.
Palien
SCHTEPA (1966) first noted thal so me species of Cousinia have the same pollen type as
Arctil/l1I. Later, she discovered that COIlSillio subgenera CYl/aroides and Hypaelllllhodes
ha ve pollen similar [O Arelil/m, while Cousillia subg. COllsinio has a different palien type
A.
SUSANNA,
N.
GARCIA-JACAS,
R.
VILATERSANA
& T. GARNATJE
105
1976). KUPRIANOVA & TSCHERNEVA (1982) conf¡rmed Ihe I\VO palien Iypes and
named them Arcl;aslrulII (cireumseribed to Arcl;//11/ and Co//sinia subg. CYllaroides and
Hypaconthodes) and Co//sinia (Iimited to COllS;Il;O subg. Co//sin;o). OUISTERMAr\T (1996)
arrived al the same eonelusions of SCHTEPA (1976) and KUPRIANOVA & TSCHERNEVA (1982).
Nevertheless, she did not favour the inelusion of alllhe speeies of lhe Co//sinia subgenera
wilh ArcliaSlrum pollen type in ArctiuIII, beeause palien types in the tribe are usually round
in more than one genus. Aceording lo our observations, SC/¡l1Ialllllllsenia has Arct;ostrum
poli en type, and the palien type of Hypacalll/¡;IlI/1 is unknown.
(SCHTEPA.
Geographic distribution
The genus ArClium has a peculiar dislribulion, with lwo groups wilh marked differences
in this respeee In lhe definition by DUISTERMAAT (1996), the species of ArCI;1l1ll sensu striclo
(sect. Arc!illm) are subeosmopolitan. The resl of the species (Le., alllhe species previously
e1assified in the genus COllsillia) have a cenlral Asian distribution, as lhe gcnus Co//sinia.
On the olher hand, distribulion of the genus COllsinia is eharaeleristie of lhe mounlain
flora of lhe Irano-Turanian Region, coincidenl wilh lhe "Orienlal·Turanian Flora Region"
defined by MEUSEL et al. (1965); see KNAPP (I987). There are two main cenlres of
speciatíon: lhe western region with 251 species, and lhe eaSlern region with 340 species
(TSCHERNEVA, 1974). Mosl of the speeies of Co//s;/Iia subg. CYlloroides and Hypacanthodes
grow only in lhe easlern regíon, as do the genera Schmalhallsellia and Hypacallt/¡;IlII1.
Only three out of se ven sections of subg. CYllaro;des are present in the western region
wilh only one species endemíc lO Iran (RECHINGER, 1972, 1979). The other species are
present in both regions. Regarding CO/lsi1l;o subgenus Hypacolllhodes, lhere are no
representatives in the western region.
Our objectives
The combined analysis of the nrONA region ITS and the chloroplast ONA gene marK
has pro ven an excellent tool for unravelling problems at this level in lhe Cardueae
(GARCIA-JACAS et aL, 2001, 2002). Thereafter, \Ve have carried out a molecular survey of
bolh regions in a sample of aH lhe genera of lhe ArctiulII group \Vith the following goals:
1) To verify lhe monophyly of the ArctiuIIl group
2) To establish the generic boundaries of Arcl;ulII and CO/lsi/l;a
3) To compare nuelear ribosomal and chloroplasl DNA phylogeny and lhe evolulion
of lhe palien types, basic chromosome numbers and lTlorphology in lhe group
4) To contribute with a preliminary study lO lhe infrageneric classificalion of
the genus COllS;ll;a.
MATEIlIALS ANO METHOOS
Plant material
Sampling was based on lhe seclional c!assificatioll of lhe genus CO/lsillio by
TSCHERNEVA (l988b, 1988c). We have inc1uded 27 taxa of the genus COl/sillia from 19
seclions, lhree species of the genus Arcliul1I, one species oC lhe genus Hypacalllllium
and the only species of the monospecific genus Sc!lmalllOlIsellio. The five outgroup
species were chosen in the genera Juri/leo and Saussureo according 10 previous
106
COLLECTANEA
BQTANICA (BARCELONA)
26. 2003
sequence analysis (SUSANNA et al., 1995; OARCIA-JACAS el al., 2002). Voueher data,
source and GenBank sequence accession numbers of the lTS and malK sequences for
(he 37 s[udied species are given in Table 1.
The analysis sequences used published sequences aJong with new sequences. ITS
sequences of Arel;"", Jappa L., Arctiu11I 111;''''5 Bernh., COllS;lI;a canescells DC.. C.
esfalldiarii Rech. f. & Aellen (both ITS and marK) and C. oflopordioides Ledeb. were from
a previous sludy (GARCIA-JACAS el al.. 2002). Sequence of COIIs;lliQ pterocaulos (C. A.
Mey.) Reeh. f. was from HilFFNER & HELLWIG (1999).
DNA Extraclion, Amplific31ion and Sequencing
TOlal geoomie DNA was extraeled following the CTAB melhod of DOYLE and DoyLE
(1987) as modified by SOLTIS et al. (1991) and CULLlNGS (1992) from siliea gel dried lea ves
collected in the field, or fresh ¡caves of plants cultivated in the Botanic Institute of
Barcelona. In sorne cases, herbarium material was used.
cpDNA malK gene strategies
Double-stranded DNAs of matK were amplified by PCR with tmK-71OF (JOHNSON &
SOLTIS, 1995) and AST-I R {OARCJA-JACAS el al., 2002} as PCR primers. Sometimes this
combinalion failed lo generate double-stranded products for some taxa and we used matKl848R (JOHNSON & SOLTIS, 1995) primer, as a subslitUle of primer AST-lR. 40 eycles of
amplification were carried OUl under lhe following conditions: 94°C for 1 minute 30
seconds, 480C for 2 minules and 720C for 3 minules, with an addilional eXlension step of 15
minutes at 72°C.
The double-stranded PCR produets were c1eaned using QlAquiek PCR Purifieation Kit
(Qiagen lne.) and sequeneed. Sequeneing primers tmK-7IOF, matK-1848R and AST-lR
were used. Oirecl sequencing of lhe amplified ONA segments was performed using a
BigDye Terminator Cycle Sequeneing v2.0 (PE Biosystems), following the protoeol recommended by the manufacturero The nucleotide sequencing was performed al the "Serveis
Cienlífieo-Téenics" of the University of Barcelona on an AB] PRlSM 3700 DNA Analyzed
(PE Biosystems). Nucleolide sequences of /l/a/K were edited using Chromas 1.56
(Teehnelysium Pty Ltd) and easily aligned by hand.
nrONA ITS region slralegies
Double-slranded DNA of lhe lTS region was amplified using the l7SE as forward primer
and the 26SE as reverse primer (SUN et al., 1994). The profile used for amplifiealion
included a warm start at 94°C for 2 minules, followed by 800C for 5 minutes, during which
the polymerase {Ecolaq, Eeogen S. R. L.} was added. 30 eyeles of amplifieation were
carried out under the following condilions: 94°C for 1 minute 30 seconds, 57°C for 2
minutes and 72°C for 3 minutes, with an additional extension slep of 15 minutes at 72°c'
The PCR produets were purified wilh the QlAquiek PCR Purification Kil (Qiagen lne.).
Both strands were sequenced with the sequencing primers 17SE as forward primer, and
26SE as reverse. Direct sequencing of lhe amplified DNA segments was performed as for
lhe matK region.
Phylogenetic analysis
DNA sequences were aligned visually by sequential pairwise comparison (SWOFFORD
& OLSEN, 1990). Data matrices are available on request from the correspondence author.
Parsimony analysis in volved heurislic searches conducted wilh PAUP version 4.0b4a
A. SUSANNA, N. GARCIA-JACAS, R. VILATERSANA & T. GARNATJE
107
TabJe 1.- Origin of the material s and herbaria where lhe vouchcrs are deposited.
Species
Arctil/III lappa L.
Arctil/lII lejosperllllll1l
Juzo & C. Serg.
ArctiulIl mjnlls Bernh.
Cousillia cllrysall1ha Kult.
COllsinia piprocepJwla Bunge
Voucher
Arctillltl
GARCIA JACAS el al. (2002)
Kazakhstan. Suscu/lla 2/54 el al. (BC)
GARCIA-JACAS el al. (2002)
COllsinia subg. COllsillia
COlIsinia secl. Alpillae
Kazakhstan, SlIsamw 2198 er al. (Be)
COl/sil/ia sect. Badgl1ysia
Iran, K. H. Rec/lillger 46730 (B)
COl/sillia riallshallica Kuh.
cOI/!.illia secl. Cardll/lCelllls
Kazakhslan, Sllsalllw 2191 el al. (BC)
Cousillia dis.fecla
COlIsinia sect. Chrysoptera
Kazakhslan, SusamUl 2137 el al. (BC)
Kar. & Kir.
COl/sinja coronala Franch.
COllsillia sect. CorollopllOra
Uzbekistan. SusaJllla 2039 er al. (BC)
COllsillia cOllgesta Bunge
COlIsillia secl. COlIsillia
Uzbek¡stan. SlIsalllUl 2059 el al. (BL)
Cousillia millkwitziae Bornm. Kazakhstan. Sl/:wmUl 2183 el al. (BC)
COl/sil/ia po/yceplwla Rupr.
Kazakhstan. SlIsamla 2161 el al. (BC)
COllsinia syrdariensis Kult.
Kazakhslan, SII.WiIllIa 2159 el al. (BC)
Accession
lIlatK
1TS
AF319048 AY013520
AF319102
AY373720
AY373687
AF319049 AY013521
AF319103
AY373725 AY373660
AY373692
AYJ737J6
AY373703
AY373743 AY373671
AY373710
AY373728
AY373695
AY373727 AY373662
AY373694
AY373726
AY373693
AY373735
AY373702
AY373738
AY373705
AY373741
AY373708
AY373óM
AY373661
AY373668
AY373669
COllsillia sect. CYllal'Oideae
COl/sillia canescens OC
GARCIA-JACAS et al. (2002)
COlIsillia ollopon/ioides
GARCIA-JACAS et al. (2002)
Ledeb.
COllsillia purpurea
Armenia. K. Ttmwllia/l (ERE)
C. A. Mey
COllsillia caespilosa
COl/si"ia sect. EriocolIsinia
Kazakhstan, SlIsamla 2170 et al. (BC)
C. Winkl.
AF319068
AF319122
AF319070
AF319124
AY373739 AY373663
AY373706
AY373724 AY373673
AY373691
COlIsinia eriobasis Bunge
COlIsinia sect. Lacflllosplwerae
Iran, K. H. Recllinger 47012 (B)
Cousillia aSlracallica
COlIsinia secl. Leiocaules
Kazakhstan, SlIscullla 2104 et al. (BC) AY373723 AY373670
(Spreng.) Tamamsch.
AY373729
AY373696
AY373690
COLLEcrANEA BOTAN1CA (BARCELONA) 26. 2003
108
COl/sinia sect. Microcarpae
KazakhSlan. Susam" 214U el al. (Be)
COlls;n;a aroe/moldea
Fisch. & C. A. Mey
COlisi"ia microcarpa Boiss.
Kazakhstan. SllsamlO 2160 el al. (Be)
COI/sil/la platylepis Schrenk
Kazakhstan.
SUSlIIlIlO
2J58
el
al. (BC)
COl/sil/¡a sewerzowii Regel
Kazakhslan,
SUSllIlIID
2178
el
al. (Be)
LOlIsillia eS{Ondiarii
UARCIA-JACAS el al. (2002)
AY3~~7n AYJ7jbOb
AY373689
AY373734 AY373667
AY373701
AY373737 AY373665
AY373704
AY373740 AY373674
AY373707
COlIsillia sect. Srenoceplwlae
Rech.
r.
& Aellen
COIIS;";O PfUOCOlllos
AF319vo9 AYOl3537
AF319123
COlIsinia sect. Xiphiolepides
HAFfNER & HEllW¡G (1999)
(C. A. Mey.) Rech. f.
Cousilllo Karoravlea
COllsinia subg. Cy"aroides
COl/sif/ia sect. Chrysis
KazakhSlan. SusamlO 2/62 el al. (BC)
Regel & Schmalh.
AY373732 AY373678
AY373699
COllsilli(1 secL ulppaceae
COl/sil/io lappacea Bunge
Kazakhstan, SI/samia 2J50 et al. (Be)
COl/sil/ia triflora Schrenk
Cousilllo secl Oligamha
Kazakhslan. SI/samia 2157 el al. (BC)
~'ollsillia
COlIsillia secl Peclillalae
Kazakf1stan. SIlSa1l1la 2200 el al. (He)
albeni
Regel & Schmalh.
AY373733 AY373677
AY373700
AY373744 AY373675
AY373711
AY~~~lll
AY:l7JbHU
AY373688
COlIsillia I/mbrosa Bunge
COllsillia sect. Pselltlarclilllll
Kazakhstan. SlIsl/I/IIa 2/00 elaf. (BC)
COl/sillia grtl1ltli/ofia Kult.
COllsitlia subg. Hypacatllhodes
COllsillia secl. Amberbopsis
Kazakhslan. SI/Sl/lllla 218/ el al. (Be)
AYJ7J745 AY37367b
AY373712
AYJ'J':~
AY37367Y
AY373697
Hypaca"thium
Hypacamll iwn ecJrinopijolium Kirguizistan. llji" (LE)
(Bomm.) Juz.
Schlllalhausellia llitlllla1lS
Schmall,allsellia
Kazakhstan. ::iusalllra 2088 el at. (BC)
(Regel) Petr.
JI/rillea albiClllllis Bunge
Outgroup
LJreece, SIISlIllIW 1957 el al. (I:R":)
JI/rillea ltlllipes Rupr.
Kazakhstan. SIISl/I/IIlI 2136 tN al. (BC)
Jurillltll robllsUl Schrenk
Kazakhstan. SUSlIllf1ll2103 el a/. (BC)
Smusllrell elega1rs Ledeb.
Kazakhslan. SI/salllla 2/79 el al. (BC)
Sallssllna maximolViczii
OfuRa BOlanieal Garden (Japan)
Herder
AY37374b
AY373713
AY373752 AY373681
AY373719
AY J w ",
AY373714
AY373748
AY373715
AY373749
AY373716
AY373750
AY373717
AY373751
AY373718
AY373"".
AY373686
AY373685
AY373683
AY373682
A.
SUSANNA.
N.
GARCIA-JACAS,
R.
VILATERSANA
& T.
GARNATJE
109
(SWOFFORD, 1999) using TBR branch swapping wilh character states specified as unordered
and unweighted. The indels were coded as fiflh base. AH most-parsimonious trces (MPTs)
were saved. To locate other potential islands of most-parsimonious trces (MADDISON,
1991), we performed 1000 replications with random taxon addition, also with TBR
branch swapping. Bootstrap analyses (BS) were performed (FELSENSTE1N, 1985) wilh
1000 replicates, and decay indices (DI) were calculated (BREMER, 1988; DONOGHUE el al.,
1992) to obtain estimates of support for each monophyletic group. AH the decay analyses
were conducted using the elade-constraint approach as discussed in MORGA N (1997).
Three parsimony analyses were performed, with three different data sets: the ITS dala, the
mQtK data and Ihe combined ITS and mal K data.
The nrDNA ITS and cpDNA matK data sets were tesled for congruence using the
partition homogeneity test (FARRIS et aL, 1995) as implemented in PAUP 4.0b4a, before
combining the data sets. The partition homogeneity test was conducted with 1000
replicates, heuristic search option with simple addilion sequence, TBR, and MULPARS.
RESULTS
Nuclear ribosomal DNA ITS
The ITS 1 and ITS 2 alignment of 37 laxa consisted of 484 positions and eontained 131
phylogenetically informative substitutions and 7 phylogenelically informative indels. Mean
pairwise distances (as calculated by PAUP) within ingroup varied from 0% (between
COlIsillia cOllgesla Bunge and C. caespirosa C. Winkl.; C. syrdariellsis Kult. and C.
polycephala Rupr.) to 11.8% (between C. kararavica Regel & Schmalh. and C. plerocalllos).
Mean pairwise distan ces between ingroup and outgroup varied rrom 10.9% (belween
Saussurea elegans Ledeb. and Hypacall1hiwll echinopifofium Juz.) to 16.8% (bclween
lurillea albicalllis Bunge and COlIsinia syrdariellsis).
The parsimony analysis yielded 91 MPTs of346 steps in one island. Thc strict conscnsus
of aH the trees is shown in Fig. 1; the consistency index (el) exeluding uninformative
eharaeters was 0.4780; the retenlion index (RI) was 0.7570; and the homoplasy index (HI)
was 0.5220.
The striet consensus of lhe 91 MPTs produeed from lhe ITS analysis (Fig. 1) SUPP0rl
monophyly of the Arclium group: Arclilllll, COlIsil/ia, Hypacanrhium and Schlllallltlusellia
(BS = 93%, DI = 7). Within this elade, there were t\Vo branches: a first Dne, which \Ve \ViII
name the "Arctioid" e1ade, comprised the genera Arctillm, Hypacal/lhiulII and
Schmalhallsellia and the representatives of COl/sillia subg. CYllaroides and HypacclIIlhodes
(SS 76%, DJ 2); und a second one formed by the represenlulives of COlIsinia
subgenus Cousinia (SS = 86%, DI = 2). Within lhe "Arclioid" elade, there were two
elades; one was formed by lhe genera Arclillm, Hypacal/lhillm and Schlllalhausellia and
COllsinia gral/difolia Kult. from subgenus Hypacalll/wdes (SS =94%, DI = 3); lhe other
e1ade was formed by the representatives of COlIsil/ia subgenus CYllaroides, with very
strong supporl (BS = 100%, DI = 11).
=
=
Chloroplast DNA malK
The mal K alignment of 27 taxa consisted of 985 posltlons and contained 21
phylogenetically informative substitutions, Mean pairwise distances (as calculaled by
PAUP) within ingroup varied from 0% (between Cousinia polycephala, C. syrdarienis and
C. millkwirziae Bomm.; C. cae~pi[osa and C. asrracanica (Spreng.) Tamamsch.; C. gral/di/ofia
110
COLLECfANEA BQTAN1CA (BARCELONA) 26. 2003
and C. karalavica) to 1.3% (between Schmal"allsell;a "idll/mls (Regel) Pelr. and COllsinia
es/ondiarjj). Mean pairwise distances between ingroup and outgroup varied from 0.3%
(between }Ilrillell robusta Schrenk and Cousillia caespitosa) to 1.3% (between Jur;nea
lanipes Rupr. and SchmaJhausenia nidulalls).
The parsimony analysis yielded 11431 MPTs of 25 steps in one island. The stricl consensus
of all the lrees is shown in Ag. 2; the consistency index excluding uninformative characters (Cl)
was 0.6333; lhe relention index (RI) was 0.8000; and lhe homoplasy index (HI) was 0.3667.
The striet consensus of lhe 11431 MPTs produced from (he marK analysis (Fig. 2)
supports monophyly of lhe Arctil/m group: Arclilllll. COlIsinia and Schlllalhausenia
(SS = 78%, DI = 2). Wilhin lhis group, lhe only well defined clade was formed by COl/si"ia
lappacea Bunge as sister (SS = 71 %, DI = 1) af a darle formed by (he genera ArcfiuIII,
Sc¡'ma'hausetl;a and lhe represenlatives of COllsinia subgenera Cynaroides and
Hypacolll/¡odes (SS = 92%, DI; 2). The reSl was unresolved as a polilomy.
Combine<! nrONA ITS and cpONA lI/olK
The P-value resulting fram the partition homogeneity test (P = 0.0570) ¡ndieates (hat data
partitions are random, and there is congruence between nrONA ITS and cpDNA IIIatK data
seIS, al a significance lhreeshold of P = 0.05 (FARRIS el al., 1995).
The combined ITS-matK alignment of 29 laxa consisted of 1469 positions wilh 149
phylogenetically informative substitutions and 6 phylogenetically informative ¡ndels. Mean
pairwise distances (as calculaled by PAUP) wilhin ingroup varied fram 0% (belween
Cousinia syrdariellsis and C. polycephala) la 3.9% (between COllsinia cOI/gesta and
ArCliulII lIIillllS). Mean pairwise distances beLween ingroup and oUlgroup varied from 4.1 %
(between Sallssllrea efegalls and COllsillia microcarpa Boiss.) to 6% (between Jurinea
afbicalllis and Sclullafhallse"ia n;dl/fans).
The parsimony analysis yielded 8 MPTs of 329 steps in one island. The strict consensus of
all the trees is shown in Fig. 3; lhe eonsistency index (el) exeluding uninforrnative charaeters
was 0.5650; Ihe relenlion index (RI) was 0.7895; and Ihe homoplasy index (HI) was 0.4350.
Like the ITS or malK alone, lhe striet consensus of the 8 MPTs obtained from the
combined analysis (Fig. 3) strongly supports (BS = 93%, 01 = 6) lhe monophyly of Ihe
Arctium group (Arctium, COlls;nia and SchmafhaIlSell;a). We have found again the two
clades of lhe ITS analysis. One clade, lhe "ArClioid" clade, was formed by Ihe genera
Aretillm, Schmafhausen;a and the representatives of COlls;nia subgenera CYllaro;des and
Hypacalll/¡odes, wilh strong support (SS = 97%, DI = 5). Wilhin lhe "Arclioid" clade, lhere
were two branehes, one with the genera Arct;llm and Schmafhallsellia, and COlIsin;a
gralldifolia from subgenus Hypacall'hodes (BS = 89%, DI = 3), and a second branch
formed by COIIsillia subgenus CYllaroides (BS = 100%, DI =12). The second clade, Ihe
COllsillia e1ade, was formed by all the representatives of Cousillia subgenus COllsi/lia
(BS:;;; 89%, DI:;;; 4). Relationships within the COlls;nia e1ade are unelear because of the
limited scope of our sampling.
D1SCUSSION
Monophyly of lhe Arctium group
The use of a combinalion of /1JalK and lTS data has proven again a good choice for our
study: the resulting tree from the analysis of the combined dala shows excel1enl resolulion
at all taxonomical levels (Fig. 3). The matK alone is not suitable for our purposes (Fig. 2)
A. SUSANNA. N. GARCIA-JACAS.
R.
VILATERSANA &
T.
GARNATJE
III
97 ,........ Aretíum lappa
94
3
Q)
. :' 76
"
2
Aretium lelospermum
5 87
~
2-
ICausínía grandífalia subg. Hyp.
-1
2"
1
72 100....2..f""P89
3
86
2
94....-
21-
G3
88 ,........
2,--
,.
o
I «...J
o'"
o~
Hyhaeanthium eehinopifoJium
Se malhausenia nidulans
Cousinia albertii
Cousinia karataviea
J,QQ.
- ,....:....r- Causinía
umbrasa [Al
11 L..;-r'i""t1 Causínía lappaeea [Al
1
Cousinia triflora
Cousinia araehnoidea
Cousinia astracaniea
,...:...
51
_ 1 74"'T'U4- Cousinia eaneseens
~
7
w
3 '-- Aretium minus
"
Ox
. ..:
f-
0>'"
O
¡)lo
«
.o§,
~
Micro x- 12
Leioc x= 12
Cynar x=
Causínía anapardíaídes Cynar x=
Causínía purpurea
Cynar x=
Causínía pteraeaulas
Xyph
Cousinia eaespitosa
Erioc x=
Cousinia eongesta
Caus x=
Causínía platylepis
Micro x=
Causínía ehrysantha
Alpin x=
Cousinia disseeta
Chrys
Cousinia minkwitziae
Caus x=
Causinía palyeephala
Cous x=
Cousinia syrdariensis
Cous x=
Cousinia coronata
Coron x=
Cousinia mieroearpa
Micro x=
Cousinia sewerzowii
Micro x=
Cousinia eriobasis
Lachn
Cousinia esfandiarii
Steno
Cousinia piptoeephala Badg x=
Cousinia tianshaniea
Card x-
12
12
12
11
13 w
11 ~ .~
...J '12 o!!l
12
12
12
13
13
11
-O>
C/J.o
:::> :>
OC/)
u
13
13
Jurinea albieauJis
100
100....- Jurínea lanipes
24 5 Jurínea robusta
(9
f-
:J
100....- Saussurea elegans
14
« o
-u
<: .
O
Saussurea maximowiezii
Fig. l. Strict consensus lree of the mosl parsimonious trees generated by Ihe ITS region
matrix. Subgenera: CYI/llr.= CYllaroides: Hyp.= Hypacall/hodes. Sections: Alpill= Alpb/(//!;
Badg= Badghysia; Card= Cardlwcelllls; Chrys= Chrysoplem; Coroll= Corollophora: COl/s=
COllsillia; CYllar= CYllaroideae; Erioe= Erioeollsi"ia; Ladll/= ulc/lIlosphael'ae: Leioe=
Leioeallles; Micro= Mieroearpae; Srello= Srel/oceplwlae; Xyp1J= Xiphiolepides. (A) = Arclioid
species. aUTG= Outgroup.
Q)'
.
.,
...)
•
Arctias/mm pollen type
~
W
COl/s;l/;a pollen type
112
COLLECTANEA BOTANICA (BARCELONA) 26. 2003
as il even fails in supporting monophyly of lhe group (SS; 78% and DI ; 2 only).
However, it is worlh remarking thallhe l1IatK analysis reflects lhe cul between Ihe "Arctioid"
and lhe Colts;llia e1ades. even with poor support (SS:;; 71%, DI:;; 1). This demonstrates
thar lhe separalion of both groups is very old, as could be expected from lhe different
pollen types and chromosome numbers.
As already suggested on lhe basis of morphoJogy, lhe Arctilllll group forro a strongly
supported monophylelie clade, bOlh in lhe ITS (SS; 93%, DI ; 7) and eombined
(SS; 93%, DI ; 6) analyses (Figs. 1,3 respeelively). This result eonfirms lhe importance
of achene characters in lhe Cardueae, as lhis group was defined mainly on Ihe basis of
shared achene apomorphies: wavy ar tigrille pattern in lhe pericarp and pappus selae
individually deciduous (SUSANNA & GARCIA-JACAS, in press).
Character evolution in the Areliu", group
Habil aud morphology
Our results suggest that the spiny habil is a derived character in the group. We can recall
lhal OARC1A-JACAS el al. (2002) plaeed lhe An:lilllll group in a clade fonned by lhe subtribe
Centallreinae and ils polential sisler groups, the genera Jurinea and Sallss/lrea. Wilhin Ihis
group, unarmed habit is overwhelmingly dominant: aH the species of Jurinea and Saussl/rea
are unarmed, and spines are a very rare fealure among the Centallre;nae: the only spiny genera
in lhe subtribe are those of the Carthamlls complex, which are a derived group (GARCIA-JACA5
el al., 200 1), a fact thal reinforces the hypothesis of the derived character of spilles. Anceslors
of the Arctillm group were probably unarmed plants lhat developed spines, like the genera of
lhe CartlulI1J11s complex, as a secondary adaptation lO the defence againsl herbivores. In lhe
case of COllsinia, spines have appeared twice in different lineages (Figs. 1,3): one in COllsin;a
subgenus COllsinia and a second one in Hypaeallthillm and Schmalhallse"ia.
Pollen and chromosomes
We shall discuss together both characters as they appear to be correlaled. As has been
repeatedly demonslraled in the e10sely related Cemallreinae, pollen and karyology are the
mosl reliable characters for phylogenetic reconstruction and usually show also a narrow
eorrelalion lo molecular phylogenies (SUSANNA el al., 1995; OARCtA-JACAS el al., 2ool). In
the Arcti",lI group. Ihe correlation is a1so very narrow and Ihey are the only ones that define
truly monophylelie groups (Figs. 1, 3).
Pollen evolulion also follows lhe panem suggested by WAGENITZ (1955) for lhe
Centaureinae, widely confirmed by molecular tools (GARCIA-JACA5 el al.. 2000, 2001).
Areliaslmm pollen type of lhe "Arctioid" e1ade (Arclillm, Sehmalhallsenia and COIIsinia
subg. Cynaroides and Hypaeanlhodes) is ancestral as compared lO the COllsi,¡ia pollen type:
Areliaslmm is spiny, very similar to Ihe anceslral Serrallda pollen Iype of the Celllallreinae,
while Cousinia pollen lype is smooth, much akin to Ihe Dealbata, Cymllls and Momana
derived pollen types of Ihe Celllallreillae (VILATER5ANA el al., 2001).
As lo chromosomal evolution, Ihe "Arctioid" e1ade is also ancestral to the Cousinia e1ade
in this character. It is generally accepted lhat higher chromosome numbers are ancestral as
compared lO lower ones, and this Irend has been demonSlrated in groups e10sely related to
the Arclillfll group like subtribe Cenlaureinae (GARCIA-JACA5 et al., 2001). In our case, the
base chromosome number x :;;; 18 of the "Arclioid" e1ade is ancestral as compared to lhe
• ; ll, 12 or 13 of lhe COIISillio clade (Figs. 1, 3).
A.
SUSANNA,
N.
GARCIA-JACAS.
R.
VILATERSANA
& T. GARNATJE
113
r - - !Aretium lappa
'"- ~rctium minus
'"- Cousinia triflora
92
2
71
1
64
1
78
2
Cousinia umbrosa
Cousinia karatavica
Cousinia grandifolia
Cousinia albertii
I-ehmalhausenia nidulans
w
O
«
...J
U
O
O
f-
U
((
«
Cousinia lappacea
Cousinia esfandiarii
Cousinia ehrysantha
Cousinia minkwitziae
Cousinia polyeephala
Cousinia syrdariensis
Cousinia coronata
Cousinia purpurea
Cousinia congesta
Cousinia
Cousinia
Cousinia
Cousinia
Cousinia
platylepis
arachnoidea
microcarpa
astracanica
tianshanica
Cousinia caespitosa
Cousinia sewerzowii
Jurinea albieaulis
Jurinea robusta
Jurinea lanipes
Fig. 2.- Strict consensus lree of (he mos! parsimonious lrees generaled by lhe
1II111K
matrix.
Generic implications
The delineation of Aretium and Cousinia
Our results pose a very inleresling but vexing problem on the generic delineation of
Aretillm and COlIsinia: how to conjugate conflicting morphological, on the one hand. and
molecular, pollen and karyological dala, on lhe olher hand.
Molecular analyses suggest a clear separation (and a very ancient one, as renected by the
matK data) between the "Arctioid" clade and COlIsinia sensu slriclo, strongly supported by
pollen and karyological dala (Figs. 1, 2. 3). The problem is, lhen, whal lO do wilh
Hypacamhium and Schmalhausenia. On strict molecular, pollen and karyological grounds,
114
COLLECTANEA BOTAN1CA (BARCELONA) 26. 2003
they belong without any doubt 10 {he "Arctioid" dade... whose homogeneity they destroy
on morphological grounds! As stated in the introduClion of this papero ¡he only useful
characlers for segregating ArClium and COltsinia are (he Icaves and, partly. (he appendages
of the bracls. Pollen types and karyology are excellent markers, hUl lhey are nol macroscopic
and henee of lillle praelieal utilily. Morphology of Ihe stigmas eould be useful (il should be
rully explored in COlls;llia s. sIr.), hUI il is very difficult lo observe too. OUT own observations
suggesl that there are sorne differences in length and distribution of (he collector hairs: lhey
are long and sparsely distributed along the whole lenglh of the stigma in COllsi/lia s. 8lT., and
short and concentrated on the basal thickening in Arctium. Schmalhausen;a is intermediate
belween bOlh models, but c10ser to Aretium. However, from our point of view, differences
are nol so substantive as c1aimed by DUlSTERMAAT (1996).
We have to resort to lhe leaves and, if we follow this morphological definition,
Hypaca1lth;1l111 and ScllIllalhal/sell;a (probably a single genus) wauld be c1assified in
COIIS;Il;a sensu stricto, conlradicting molecular, karyological and pollen evidence. There are
three possible solutions to this incongruence.
The first ane should be to keep present c1assification in four genera with a different
delimitation: Aret;ulII (ineluding COIls;llia subgenera Cynaro;des and Hypacallthodes),
COlls;1lia, Hypaca1lthillm and Schmalhal/sen;a, disregarding the fact of the polyphyly of the
resulting genus Aret;um (Figs. 1,3). This solution does nal seem adequate, if our goal is lO
achieve a natural c1assification.
A second altemative would be to elevate the subgenera of COlIs;n;a wilh Arct;aSTrlllll
palien type to the genus rank. The resulting Arct;um group would encompass six genera:
Arct;ut1l sensu str., COlls¡'lia s. str., Cy"aro;des, Hypaca1lth;um, Hypacallthodes and
Schmalhallsenia. The possible genus CY1laroides stands as a very well defined group on
molecular grounds (SS = 100%, DI = II in Ihe ITS lree, Fig. 1; SS = 100%, DI = 12 in Ihe
combined tree, Fig. 3), and our sampling is complete enough as to draw conelusions (five
oul of seven seelions of Ihe subgenus, according lO TSCHERNEVA'S (l988b, 1988e) c1assification). In contrast, our sampling of sect. Hypacallthodes is too short (one out of four
seclions) and the only included species forms a well·supported c1ade with Hypacalllh;lIm
and Sehllla/hal/senia (SS = 93%, DI = 5 in the ITS Iree, Fig. 1; SS = 90%, DI = 5 in Ihe
combined Iree [withoul Hypacallth;lIm], Fig. 3), a grouping lhal needs verificmion. However, we can anticipate that the problem of making up these six genera ¡s, again, morpho·
logical. The boundaries between Hypacalllh;u11l and Schmalhallsen;a, on the one hand, and
Cous;/I;a sensu str., on the other hand, are unc1ear: pollen type and karyology are the only
reliable characters thal separale lhem. The stigma characters could be of help, bUl further
studies are needed in COIIS;ll;a. Besides, the limits between Arct;u11I and a new genus
Cy"aroides would also be uncertain on slrict morphological grounds because subg. Cy"aroides
¡neludes the arctioid species of Cous;nia.
Finally, Ihere is Ihe solulion suggesled by KUNTZE (1891): lO lump logelher alllhe genera
described in lhe Arcliu11I group in a single genus Arctillm (as this genus would be
nomcnelaturally prioritary). We are conscious of the nomenclatural consequences of this
solution: sorne 420 species of Cousinia will have to be changed to Areti/l'" -KUNTZE (1891)
already proposed ca. 180 new combinations·. Nevertheless, if new studies in the group
(especially with the ¡nclusion of mOfe species of CO/ls;/lia subgenus Hypacallthodes)
confirm our results, it will probably be the only solution.
Molecular phyloge,ry o1ld sectional classifieat;orl o/ CousilJia
We are aware of the severe Iimitations of our sampling regarding COlIsill;a subg. Cousillia.
However, from our study emerges a general incongruence belween sectional c1assification
A.
SUSANNA.
N.
GARCIA-JACAS.
R.
VU..ATERSANA
& T.
Arctium /appa
Arctium minus
89
3
[Cousinia grandifolia Hyp
Schmalhausenia nidulans
Cousinia a/bert;;
Cyn
Cousinia karatavica Cyn
lOO
Cousinia umbrosa [A] Cyn
12
~ 1
...;;.,1
Cousinia lappacea [A] Cyn
1
Cousinía triflora
Cvn
Cousinia arachnoidea
77
Cousinia astracanica
2
Cousinia purpurea
Cousinia caespitosa
58 6
Cousinia congesta
1
Cousinia platylepis
Cousinia chrysantha
~
Cousinia minkwitziae
5 63
Cousinia po/ycephala
2
3
Cousinia syrdariensis
Cousinia tianshanica
Cousinia esfandiarii
Cousinia coronata
69
Cousinia microcarpa
1
Cousinia sewenowii
Jurínea a/bícaulis
100
27 ~ Jurínea lanípes
5
Jurínea robusta
Saussurea elegans
100
. ..
17
Saussurea maXlmOWICZII
-K
@
"
" >
"
97
5
115
GARNATJE
~
TC
w
I~
...J
üeo
o~
-"
Ox
f-
Ü
a:
«
-te
93
6
~
67
2
~
l2..
4
~
~JfC
w
O'"
:)~
üÑ
-«~
<:~
-~
"'"
;)x
O
Ü
,...
....
o.
::J
O
o::
C9
f::J
O
Fig. 3. Slrict consensus tree of the most parsimonious trees generated by Ihe combined IIIt1tKITS malrix. Subgenera: CYllar.= CYI/aroitles; Hyp.= HYPllclIIlIllOdes. (A] = Arclioid species.
\Q)'
"
':)
Arctiastrum palien type
~;
COl/sil/ia palien type
by TSCHERNEVA (1988b, 1988e), mapped in lhe ITS eonsensus tree (Fig. I" and molecular
phylogeny. By way of example. the fOUT Tepresenlatives of sect. Coltsitlia do nol fOTm a
monophylelic e1ade. Species of sect. COlls;nia with x = 12 are placed in the same elade.
whilst Colts;/I;a cOllges/a with x = 13 is placed in a different e1ade (Fig. 1). AnOlher case is
secl. Microcarpae. with four represenlatives that are placed in Ihree different eludes
116
COLLECfANEA BOTANICA (BARCELONA) 26. 2003
(Fig. 1). In bolh cases, the groups seem la be more correlated to chromosome numbers than
(O preseot c1assification. This result apeos an interesting way lO further sludies in the genus
Cousinia.
ACKNOWLEDGEMENTS
AUlhors th.nk A. 1. Iv.schenko, L. K.puslin., F. Kh.ss.nov .nd J. V.lles for their help
with field collections in Central Asia; 1. Valles again for his help with the Russian literature;
R. VOgl, Botanical Museum of Berlio, for authorising the use of herbarium material for OUT
molecular analyses; and the Ofuna Botanical Garden (Japan) for providing seeds of
Sallssurea maximowicl.ii Herder. This research was supported by (he Dirección General de
Enseñ.nz. Superior, Sp.in (projecl PB 97/1134); the Dirección General de Investig.ción,
Ministerio de Ciencia y Tecnologí., Sp.in (projects BOS200 1-304 I-C02-02); .nd the
Gener.lit.l de C.l.luny. (.juts • grups de recerca consolid.ts 1999SGROO332 .nd
200 1SGROO I25).
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