52 February 2003: 33–50
Rapini & al. Asclepiadeae classification
Asclepiadeae classification: evaluating the phylogenetic relationships of New
World Asclepiadoideae (Apocynaceae)
Alessandro Rapini1, 2, Mark W. Chase 2, David J. Goyder2 & Jayne Griffiths 2
1 Departamento
de Ciências Biológicas, Universidade Estadual de Feira de Santana, Av. Universitária s/n,
CEP 44031-460, Feira de Santana, Bahia, Brazil. rapinibot@yahoo.com.br
2 Jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3DS, U.K. m.chase@ rbgkew.
org.uk; d.goyder@rbgkew.org.uk
To provide an overview of New World Asclepiadoideae, we here evaluate Asclepiadeae classification by comparing the taxonom ic arrangement of subtribes with a topology obtained through analyses of two plastid DNA
regions (trnL intron and trnL-F intergenic spacer) for 111 species of Asclepiadoideae representing the major
lineages of the subfamily. Without Jobinia, Nephradenia and Barjonia, Asclepiadeae are not monophyletic.
The monotypic African genus Eustegia, with pendent pollinia, m ay represent the sister clade of Marsdenieae Ceropegieae, the group composed of plants with erect pollinia. Metastelmatinae including African plants are
also non-monophyletic, and the circumscription of the recently re-instated Cynanchinae should be studied further. Overall, Asclepiadeae are composed of three main clades. The Old World Astephaninae are the sister
group of the other Asclepiadeae, which are divided into the ACTG (Asclepiadinae, Cynanchinae, Tylophorinae
and Glossonematinae) and M OG (M etastelm atinae, Oxypetalinae and Gonolobinae) clades. According to this
study, the New World Asclepiadoideae fall into just four clades: (1) M arsdenia (Marsdenieae), (2) Asclepias
(Asclepiadinae, Asclepiadeae), (3) Cynanchum subgenus Mellichampia (Cynanchinae, Asclepiadeae), and (4)
MOG, the clade comprising the majority of New World Asclepiadoideae.
KEYWORDS: Apocynaceae, Asclepiadeae, Asclepiadoideae, classification, m olecular phylogeny, New World.
INTRODUCTION
Since the widespread acceptance of explicitly phylogenetic classification in botany in the 1990s,
Apocynaceae Juss. have been subject of several phylogenetic studies. The pioneering work within Asclepiadoideae R. Br. ex Burnett was carried out in Microloma
(Astephaninae Endl. ex Meisn.; Wanntorp, 1988).
Generic relationships within Astephaninae were also
investigated in morphological cladistic analyses (Liede,
1994, 1996a; Bruyns, 1999a) and recently re-investigated with molecular data (Liede, 2001). Other particular
groups, such as Sarcostemma s.l. (Liede, 1996b; Liede &
Täuber, 2000), Cynanchum (Liede, 1997a; Liede &
Kunze, 2002; Liede & Täuber, 2002), stapeliads (Bruyns,
2000), Brachystelma Sims s.l. (Meve & Liede, 2001),
Glossonematinae K. Schum. (Liede & al., 2002) and
Asclepiadinae Endl. ex Meisn. (Goyder & al., unpubl.),
have also been analysed phylogenetically (Table 1).
Broader cladistic studies including members of
Asclepiadoideae (Downie & Palmer, 1992; Chase & al.,
1993; Olmstead & al., 1993; Judd & al., 1994; Struwe &
al., 1994; Sennblad & Bremer, 1996, 2000, 2002;
Civeyrel & al., 1998; Fishbein, 2001; Potgieter & Albert,
2001) have been concerned with relationships at higher
taxonomic levels, providing little information within the
subfamily. Sampling has been so sparse (Table 1) compared to the number of Asclepiadoideae taxa that these
studies might be considered non-representative for statements regarding tribal circumscription (Endress &
Bruyns, 2000). The broadly accepted amalgamation of
Apocynaceae, Asclepiadaceae R. Br. and Periplocaceae
Schltr. has emerged as result of these cladistic analyses.
The taxonomic change of including Asclepiadaceae as
subfamily of Apocynaceae s.l., however, was due to the
adoption of a phylogenetic concept of classification,
which avoids the recognition of non-monophyletic
groups such as Apocynaceae s.str. (i.e., excluding members of Asclepiadaceae and/or Periplocaceae; Rapini,
2000). Within Asclepiadoideae, relationships are still far
from consensus (Endress & Bruyns, 2000; Endress &
Stevens, 2001).
Due mainly to the artificial circumscription of many
New World genera, which are still based on the variable
corona and misinterpretations of inflorescence characters
(Rapini, 2002), relationships among American groups
are poorly understood. New World Asclepiadoideae (Fig.
1) have not yet been the focus of any phylogenetic study,
and representation of these in analyses has been thin
(Table 1); even large genera, such as Oxypetalum
33
Rapini & al. Asclepiadeae classification
52 February 2003: 33–50
Table 1. Main phylogenetic studies including Asclepiadoideae taxa, showing number of terminals, their representation
of number of species and genera of Asclepiadoideae (Asclep.) and those in the New World (NW), as well as kind of data
used in the studies. * including trnT-L intergenic spacer.
Species
Terminals (Asclep.)
Genera
Species
Genera
(Asclep.) (NW Asclep.) (NW Asclep.)
Authors
Year
Wanntorp
Downie & Palmer
Chase & al.
Olmstead & al.
Judd & al.
Liede
Struwe & al.
1988
1992
1993
1993
1994
1994
1994
25
125
499
184
10
13
235
20
2
4
2
1
1
1
2
13
1
0
2
0
4
0
1
1
1
2
0
1
Liede
Liede
Sennblad & Bremer
Liede
Civeyrel & al.
Bruyns
Bruyns
Liede & Täuber
Sennblad & Bremer
Fishbein
Liede
Potgieter & Albert
1996a
1996b
1996
1997
1998
1999a
2000
2000
2000/2002
2001
2001
2001
15
22
29
117
46
16
35
25
79
40
43
154
31
7
116
10
25
15
29
43
50
14
12
7
4
10
15
32
14
15
21
28
36
0
22
1
1
3
0
0
12
6
13
9
9
0
4
1
1
3
0
0
5
6
7
8
7
Meve & Liede
Meve & Liede
Liede & al.
Sennblad & Bremer
Liede & Kunze
Liede & Kunze
Liede & Kunze
2001
2001
2002
2002
2002
2002
2002
24
22
29
18
91
88
64
24
21
29
1
85
86
64
13
12
22
1
11
11
4
0
0
8
0
8
8
1
0
0
8
0
2
2
1
2002
this study
108
114
107
111
31
68
36
65
13
26
Liede & Täuber
Rapini & al.
(around 130 species), have not been considered. The
study of Potgieter & Albert (2001), with 37 genera of
Asclepiadoideae, includes the greatest generic diversity
of the subfamily so far. However, that analysis has only
seven New World genera of Asclepiadoideae, and five
Asclepiadeae (R. Br.) Duby are not identified to species,
which in some cases (e.g., Cynanchum) may represent
completely unrelated taxa (Liede & Täuber, in press; see
below).
Since New World Asclepiadoideae are mainly composed of members of Asclepiadeae, the first step toward
understanding the relationships of American groups is a
good evaluation of the component subtribes. After Liede
& Albers’ (1994) tribal arrangement of Asclepiadaceae
(Asclepiadoideae, Secamonoideae Endl. and Periplocoideae R. Br. ex Endl.) genera, Liede (1997b) presented
34
Data
Morphology
Molecular (rbcL)
Molecular (rbcL)
Molecular (rbcL)
Morphology
Morphology
Morphology, anatomy,
chemistry, embryology
Morphology
Morphology
Molecular (rbcL)
Morphology
Molecular (matK)
Morphology
Morphology
Molecular (trnL-F*)
Molecular (rbcL)
Molecular (matK)
Molecular (trnL-F*)
Molecular (trnL-F),
fruit morphology
Molecular (trnL-F*)
Molecular (ITS)
Molecular (trnL-F*)
Molecular (rbcL + ndhF)
Molecular (trnL-F*)
Molecular (ITS)
Molecular (trnL-F*+ ITS),
anatomy
Molecular (trnL-F*)
Molecular (trnL-F)
a subtribal classification of Asclepiadeae. Despite that
little phylogenetic information was available at that time
and none was added in that paper, the subtribes were supposed to be defined by synapomorphies. An empirical
morphological evaluation of Liede’s classification
(Bruyns, 1999b) pointed out some inconsistencies of
subtribes. Liede (1999) replied to the criticism, suggesting the exceptions outlined might represent reversals or
parallelisms. Equally, in the absence of supporting evidence, one could argue that the similarities selected by
Liede (1997b) as synapomorphies are actually homoplasies themselves. Moreover, omission of some New
World genera (Rapini, 2002) has raised doubts concerning monophyly of the tribe itself, a point so far undisputed (Liede, 1999; Liede & al., 2002).
Since Liede (1997b) based the subtribes of
52 February 2003: 33–50
Rapini & al. Asclepiadeae classification
Fig. 1. Representative genera of Asclepiadeae. A, Ditassa cordeiroana (Brazil); B, Cynanchum roulinioides (Brazil); C,
Oxypetalum insigne (Brazil); D, Blepharodon nitidum (Brazil); E, Marsdenia suberosa (Brazil); F, Asclepias curassavica (Brazil). Photos by A. Rapini.
35
Rapini & al. Asclepiadeae classification
Aidomene parvula
Asclepias syriaca
Aspidoglossum biflorum
Aspidonepsis diploglossa
Calotropis procera
Cordylogyne globosa
Fanninia caloglossa
Glossostelma angolense
Gomphocarpus fruticosus
Kanahia laniflora
Lagarinthus tenuis
Mackenia
Margaretta rosea
Miraglossum pulchellum
Odontostelma welwitschii
Pachycarpus grandiflorus
Parapodium costatum
Pergularia tomentosa
Schizoglossum atropurpureum
Stathmostelma gigantiflorum
Stenostelma capense
Trachycalymma cristatum
Woodia verruculosa
Xysmalobium undulatum
Asclepiadinae
Ceropegia candelabrum
Stapelia hirsuta
CEROPEGIEAE
Adelostemma gracillianum
Ampelamus albidus
Biondia chinensis
Blepharodon lineare
Cyathostelma
Cynanchum acutum
Ditassa banksii
Folotsia sarcostemoides
Funastrum
Gonioanthela odorata
Graphistemma chinensis
Grisebachiella hieronymi
Hemipogon acerosus
Holostemma ada-kodien
Karimbolea verrucosa
Macroditassa adnata
Mahawoa montana
Merrillanthus hainanensis
Metaplexis stauntonii
Metastelma parviflorum
Nautonia nummularia
Orthosia congesta
Oxystelma esculentum
Platykeleba insignis
Pentacyphus boliviensis
Pentarrhinum insipidum
Pentastelma auritum
Peplonia nitida
Petalostelma martianum
Philibertia solanoides
Raphistemma pulchellum
Rhyssostelma nigricans
Sarcostemma viminale
Sichuania alterniloba
Tassadia obovata
Metastelmatinae
52 February 2003: 33–50
Astephanus triflorus
Blyttia arabica
Diplostigma canescens
Emicocarpus fissifolius
Eustegia minuta
Goydera somaliensis
Microloma tenuifolium
Oncinema roxburghii
Pentatropis microphylla
Pleurostelma grevei
Ryncharrhena atropurpurea
Schizostephanus alatus
Seshagiria sahyadrica
Tylophora flexuosa
Vincetoxicum hirundinaria
Glossonema boveanum
Odontanthera reniformis
Solenostemma arghel
Astephaninae
Fisheria scandens
Gonolobus macrophyllus
Gyrostelma oxypetaloides
Hypolobus infractus
Macroscepis obovata
Matelea palustris
Metalepsis cubensis
Metoxypetalum retusum
Pherotrichis balbisii
Schubertia multiflora
Stelmagonum hahnianum
Trichosacme lanata
Glossonematinae
Gonolobinae
Amblyopetalum coccineum
Amblystigma hypoleucum
Araujia sericifera
Diplolepis meziesii
Melinia candolleana
Mitostigma tomentosum
Morrenia odorata
Oxypetalum banksii
Schistogyne sylvestris
Stenomeria decalepis
Tweedia macrolepis
Widgrenia corymbosa
Oxypetalinae
Fockea edulis
FOCKEEAE
ASCLEPIADEAE
Secamone emetica
Periploca graeca
Periplocoideae
Asclepiadoideae
Barjonia racemosa
Cionura erecta
Dischidia nummularia
Dregea floribunda
Gymnema sylvestre
Hoya carnosa
Jobinia hernandifolia
Marsdenia tinctoria
Micholitzia obcordata
Nephradenia acerosa
Telosma odoratissima
MARSDENIEAE
Secamonoideae
Fig. 2. Tribal arrangement of Asclepiadoideae with emphasis on subtribal classification of genera of Asclepiadeae
sensu Liede (1997b; for complete generic list of other tribes, see Liede & Albers, 1994 and Liede’s homepage, www.unibayreuth.de/departments/planta2/triblist.html). Genera in bold are included in the analyses and when represented by
type species the binomial is in bold.
Asclepiadeae on supposed synapomorphies, they should,
by implication, represent monophyletic groups (i.e., the
hypothesis of phylogenetic patterns behind the classification). Our aim here is to evaluate the monophyly of
Asclepiadeae sensu Liede (1997b) and its subtribes, summarising the phylogenetic information available for
Marsdenieae Benth. and Asclepiadeae to establish an
overview of New World Asclepiadoideae as a basis for
further studies.
We analysed DNA sequences of two plastid regions,
the trnL intron and trnL-F intergenic spacer (hereafter
trnL-F). Previous phylogenetic studies (Table 1) have
often used trnL-F in molecular systematics of Asclepiadoideae, and to include a greater diversity of Asclepiadeae and Marsdenieae, the two tribes natively represented in New World, we selected representatives of most
genera with sequences published, also adding new
sequences (no published study has included all of these
data).
36
MATERIALS AND METHODS
We included sequences of 115 species in 69 genera
of Asclepiadoideae; of these, 65 species of 26 genera represent New World taxa. Fifty-four species were sequenced for this study and the remaining 61 species retrieved
from either GenBank (www.ncbi.nlm.nih.gov) or Liede’s
homepage (www.uni-bayreuth.de/departments/planta2/
wgl/fsigrid2.html; Appendix 2). We have thus included
the three genera of Glossonematinae (3), more than 50%
of genera of Asclepiadinae (13), Astephaninae (11) and
Metastelmatinae Endl. ex Meisn. (21), a third of genera
of Oxypetalinae Decne. (4; we considered Mitostigma
parviflorum Malme a member of Melinia and Tweedia
coerulea D.Don ex Sweet a member of Oxypetalum),
25% of genera of Gonolobinae (G. Don.) Liede (3; we
considered Metalepis albiflora Morillo a member of
Cynanchum) and Marsdenieae (11), two genera of
Ceropegieae Orb. (2) and Fockea edulis representing
Fockeeae Kunze, Meve & Liede. Apocynum L.
(Apocynoideae Burnett), Periploca L. (Periplocoideae),
and Secamone R. Br. (Secamonoideae) were used as out-
Rapini & al. Asclepiadeae classification
52 February 2003: 33–50
a
63
52
Stenostelma corniculatum
b
55
Gomphocarpus fruticosus
Asclepias curassavica
Stathmostelma gigantiflorum
Stenostelma corniculatum
Calotropis procera
Schizoglossum alpestre
55
Kanahia laniflora
Pachycarpus spurius
Stathmostelma gigantiflorum
62
Aspidoglossum ovalifolium
Schizoglossum alpestre
87
Pachycarpus spurius
Glossostelma spathulatum
Xysmalobium undulatum
Margaretta rosea
99
Aspidoglossum ovalifolium
Gymnema sylvestre
55
95
Asclepias curassavica
Glossostelma spathulatum
Xysmalobium undulatum
Kanahia laniflora
75
Calotropis procera
Pergularia daemia
Pergularia daemia
Dregea sinensis
65
65
Marsdenia suberosa
58
Micholitzia obcordata
51
Telosma cordata
Micholitzia obcordata
62
93
Hoya australis
100
100
Stapelia glanduliflora
Ceropegia saxatilis
81
Dregea sinensis
Dischidia bengalensis
Stapelia glanduliflora
Hoya australis
Dischidia bengalensis
96
Gymnema sylvestre
62
Margaretta rosea
Gomphocarpus fruticosus
62
Telosma cordata
Ceropegia saxatilis
Marsdenia suberosa
Fockea edulis
Fockea edulis
Fig. 3. Incongruent results between the two pieces of trnL-F from Gymnema sylvestre. Numbers above branches show
bootstrap percentages. a, position of G. sylvestre according to trnL intron. b, position of G. sylvestre according to trnLF intergenic spacer. Black bar: the Marsdenieae-Ceropegieae clade (with erect pollinia). White bar: Asclepiadeae
(Asclepiadinae, with pendent pollinia).
group (Civeyrel & al., 1998; Sennblad & Bremer, 2000;
Potgieter & Albert, 2001). Type species of each genus
were used when available (Fig. 2).
Gymnema sylvestre was not included in general
analyses because analyses of the general matrix raised
suspicions about G. sylvestre sequences. To verify a possible disagreement between the two regions of trnL-F,
we analysed each region independently. Fig. 3 shows
bootstrap (gap coding included, no region excluded)
trees of matrices (available upon request from MWC)
including G. sylvestre, representatives of Ceropegieae
and Asclepiadinae (Asclepiadeae); Fockea, sister group
of the rest of Asclepiadoideae (see below), was used as
outgroup (see Table 1 for accession numbers). According
to trnL intron, G. sylvestre is nested in Marsdenieae–
Ceropegieae clade (Fig. 3a), whereas using trnL-F intergenic spacer the species nests in Asclepiadinae
(Asclepiadeae) (Fig 3b). In each case the position of G.
sylvestre is supported by a high bootstrap percentage,
indicating that its combined trnL-F may represent a
hybrid sequence. Although trnL-F intergenic spacer represents less than 25% of the total sequence of these three
regions (trnT-L intergenic spacer, trnL intron and trnL-F
intergenic spacer), the use of this sequence of G. sylvestre
as an outgroup may disturb analyses and might, for
instance, explain the basal polytomy in Liede & Täuber
(2000).
DNA was extracted from either material dried in silica gel (Chase & Hills, 1991) or herbarium collections
(Savolainen & al., 1995, and references therein) using
2X CTAB (Doyle & Doyle, 1987). Total DNA was
cleaned with QIAquick columns (Qiagen, Inc.) using
manufacturer’s protocol for PCR products. Primers “c”
and “f” (forward and reverse, respectively) were used for
PCR of DNA in good condition; in cases where the DNA
was not intact, primers “c” and “d” as well as “e” and “f”
(Taberlet & al., 1991) were combined (forward and
reverse, respectively) to amplify trnL-F in two pieces.
For amplification, 3 min. at 94°C preceded 28–32 cycles
of denaturation at 94°C for 1 min., annealing at 48–50°C
for 1 min. and extension at 72°C for 2 min., which was
followed by more 2 min. of extension at 72°C. PCR
products were purified with Concert Rapid PCR
Purification System (GibcolBRL) and used as template
for cycle sequence reaction with the same primers used
for PCR. Sequences were processed on a 377 DNA
sequencer (ABI Prism) following manufacturer’s protocols (Applied Biosystems, Inc.; ABI).
Sequences were first edited in Sequence Navigator
(ABI) and the two complementary strands overlain in
AutoAssembler version 1.4 (ABI) for further edition.
Alignment was carried out manually following the
guidelines of Kelchner (2000), and we minimised the
number of gaps at the expense of substitutions. Twenty
37
Rapini & al. Asclepiadeae classification
parsimony-informative gaps longer than two base pairs
were coded according to the “simple indel coding” (presence/absence; Simmons & Ochoterena, 2000).
Ambiguous regions were excluded from the analyses
(matrix available upon request from MWC).
Bayesian analysis was conducted in MrBayes
(Huelsenbeck & Ronquist, 2001) using four Markov
chains simultaneously started from random trees. Two
evolutionary models were used: a simple two parameter
(transition-transversion) substitution model (nst = 2,
rate = equal and basefreq = equal) and the general timereversible model with gamma variation (nst = 6,
rate = gamma and basefreq = estimate). Five hundred
thousand cycles were performed with each model, sampling a tree at every 10 generations. Trees that preceded
the stabilisation of the likelihood value (the burn in) were
excluded, and the remaining trees were used to construct
a consensus in PAUP (version 4.0; Swofford, 2001).
Parsimony analyses were also carried out using
PAUP. One thousand replicates of random taxon addition
and tree-bisection-reconnection (TBR) branch swapping
were performed, saving no more than 10 shortest trees
per replicate to minimise swapping of suboptimal
islands. Trees in memory were then used in another
round of TBR swapping. To avoid an indefinite search, a
limit of 20,000 trees was set. This procedure was carried
out with (matrix 1) and without coding of the gaps
(matrix 2). Support for clades was obtained with 1,000
bootstrap replicates with simple addition and TBR swapping, saving no more than 10 trees to minimise swapping
on large islands.
RESULTS
The two models used for Bayesian analyses gave
nearly identical results; changes in topology were
restricted to clades with less than 50% probability, and
variation in probabilities of clades with high confidence
(above 94%) was up to 6%. We have opted for presentation of the consensus tree produced with the simplest
model. Stabilisation of likelihood values was reached at
about 100,000 replicates. Ten thousand trees were therefore discarded (the burn in) and the remaining 40,000
trees were used in estimating the probability of clades.
Matrix 1, with 179 potentially parsimony-informative
characters, generated most parsimonious trees of 594
steps, consistency index (CI) 0.72 and retention index
(RI) 0.86, whereas matrix 2, with 159 potentially parsimony-informative characters, generated shortest trees
with 563 steps, CI 0.72 and RI 0.85. The two matrixes
each reached the limit of 20,000 trees. The strict consensus tree generated by matrix 1 differed from that generated by matrix 2 but did not contradict it. The analysis of
matrix 1 gave less resolution within MOG (Metastelm38
52 February 2003: 33–50
atinae, Oxypetalinae and Gonolobinae) clade, but was
more informative concerning basal nodes, including the
dichotomy between ACTG [Asclepiadinae, Cynanchinae
K.Schum., Tylophorinae (K.Schum.) Liede and
Glossonematinae] and MOG clades as well as the
Glossonema–American Cynanchum clade. Gaps were
important in providing evidence for particular groups,
mainly in Asclepiadinae and the Glossonema–American
Cynanchum clades (Figs. 4 and 5).
In Asclepiadoideae (Figs. 4 and 5), Fockea edulis
comes out as sister to the rest of Asclepiadoideae.
Overall, the dichotomy between plants with erect
pollinia, namely the Marsdenieae–Ceropegieae clade
(100% posterior probability/88% bootstrap with matrix
1), and Asclepiadeae (99/61), with pendent pollinia, are
present. Eustegia minuta, a species with pendent pollinia,
however, appeared as sister to the plants with erect
pollinia, a relationship with high posterior probability
but weakly supported by the bootstrap (98/50). In
Asclepiadeae, Astephaninae sensu Liede (2001) received
100/100 and appeared as sister to other Asclepiadeae,
which are divided into two clades, ACTG (95/<50) and
MOG (100/69). Although this latter dichotomy is not
supported by the bootstrap, it is present in the strict consensus tree of matrix 1 and confirmed by Bayesian analysis.
In the ACTG clade (Figs. 4 and 5), Asclepiadinae
sensu Liede (1997b; 93/84) and Tylophorinae (100/83),
as well as the American Cynanchum (100/78) and the
clade composed of Glossonema, Odontanthera and
Pentarrhinum (100/87), groups already observed in
Liede & Täuber (2002), are confirmed. The clade composed of American Cynanchum, the Glossonema clade
and Metaplexis (67/86), even though with low probability according to Bayesian analysis, had a great improvement in support with the inclusion of gap information
(matrix 1). The clade comprising Sarcostemma and the
stem-succulent, cynanchoid genera Folotsia, Karimbolea
and Platykeleba (Liede & Meve, 2001) are supported
(100/68). However, Cynanchinae, as circumscribed by
Liede & Täuber (2002), are not supported, and the positions of Schizostephanus, Oxystelma and Solenostemma,
as well as the relationship among subtribes are not
resolved within the ACTG clade as a whole.
In MOG (Figs. 4 and 5), the positions of the South
American “Astephanus” geminiflorus and “Cynanchum”
nummulariifolium, as well as the monotypic Grisebachiella, are not resolved. Metastelma scoparium, with
a controversial placement (Rapini, 2002), is closely related to Orthosia urceolata (100/74). Jobinia is sister group
of the Orthosia urceolata–Metastelma scoparium clade
(100/71), and together they form a clade with
“Cynanchum” morrenioides and “Ditassa” subtrivialis
(87/<50). The main subtribal components are, however,
52 February 2003: 33–50
included in the MOG core group (100/<50), a clade present in both parsimony analyses. Gonolobinae (100/96)
are well supported, Oxypetalinae (95/<50), including
Philibertia, are also present in both parsimony analyses,
and Metastelmatinae (100/54) even though with high
posterior probability are not present in the strict consensus tree of matrix 1. The erect, usually caespitose species
of Ditassa (D. acerosa, D. decussata, D. ditassoides, D.
grazielae and D. magisteriana) appear together (100/80),
and D. cordeiroana is related to Petalostelma sarcostemma (93/65). Funastrum and Tassadia are not resolved in
MOG core, and, as for ACTG, relationships among the
component subtribes are unclear.
DISCUSSION
Fockeeae, including Cibirhiza Bruyns with two
species and Fockea with around five species, are sister to
other Asclepiadoideae, which agrees with previous phylogenetic studies (Civeyrel & al., 1998; Sennblad &
Bremer, 2000; Fishbein, 2001; Potgieter & Albert, 2001)
and also morphological evidence (Kunze, 1993; Kunze
& al., 1994). Asclepiadoideae with erect pollinia form a
monophyletic group. Due to the under-sampling of the
Marsdenieae–Ceropegieae clade, however, the monophyly of Marsdenieae cannot be properly evaluated in
this study (see Potgieter & Albert, 2001, for a broader
molecular analysis of this group, and Bruyns & Forster,
1991, and Swarupanandan & al., 1996, for discussions of
morphological characters).
The rare, monotypic Eustegia, with a three-seriate
corona and pendent pollinia (Bruyns, 1999a), has an
unexpected position as sister to the taxa with erect
pollinia (Fig. 4). Previously, pendent pollinia were treated as the ancestral state (Schumann, 1895). Currently,
erect pollinia have been considered the plesiomorphic
form and the pendent orientation an apomorphy of
Asclepiadeae (Wanntorp, 1988; Kunze, 1993; Endress &
Stevens, 2001). Few phylogenetic studies have been used
to clarify this point, and studies on Asclepiadeae using
members of Marsdenieae as outgroup (e.g., Liede, 2001)
are unable to conclude anything concerning orientation
of pollinia (contra Endress & Stevens, 2001). Some characters of Eustegia, e.g., the frail corpuscle with V-shaped
profile floor and elaborate corona (Bruyns, 1999a), may
represent plesiomorphic states shared with Fockeeae.
Others, such as absence of a narrow neck between ovary
and style head in mature flowers, are shared with taxa of
Ceropegieae (Bruyns, 1999a). Although Decaisne (1844)
included Eustegia and Fockea in the same “subtribe” (as
division of a tribe), a relationship between Eustegia and
taxa with erect pollinia has never been previously suggested due to the clearly pendent pollinia.
Rapini & al. Asclepiadeae classification
If Eustegia minuta is confirmed as sister to the
Marsdenieae–Ceropegieae clade, Asclepiadeae may then
be divided into three main clades. Astephaninae, with
100% support, are sister to the clade that comprises the
rest of the tribe, which is divided into the ACTG and
MOG clades. Even though not supported by bootstrap in
our analysis, the dichotomy between ACTG and MOG
was supported by Bayesian inference and also the bootstrap when trnT-L intergenic spacer has been included
(Liede, 2001; Liede & Täuber, 2002). MOG is restricted
to the New World, whereas ACTG, despite having two
New World groups, Asclepias and Cynanchum subgenus
Mellichampia (A.Gray ex S.Wats.) Woodson, is centred
in the Old World. Relationships among their components
are still unresolved, but this dichotomy is significant for
subtribal delimitation.
Comparing the topology obtained from molecular
data with that based on Asclepiadeae classification sensu
Liede (1997b), disagreements are evident (Fig. 4; Table
2), and several changes have been made mainly by that
author to improve the classification (Fig. 5). Astephaninae have been reduced to just three genera, Astephanus,
Microloma and Oncinema, a group well supported by
both morphological analysis (Liede, 1994, 1996a; but see
Bruyns, 1999a) and molecular data (Liede, 2001). The
remaining genera (Blyttia, Diplostigma, Goydera,
Pentatropis, Pleurostelma Baill., Ryncharrhena F.
Muell., Tylophora and Vincetoxicum) together with
Biondia form a well-supported group now considered as
Tylophorinae (Liede, 2001). The New World genera
Barjonia, Nephradenia and Jobinia, previously included
in Marsdeniinae K.Schum. (nom. superfl.) or Marsdenieae (e.g., Schumann, 1895, and Liede & Albers,
1994, respectively), have also been considered in
Asclepiadeae (Endress & Bruyns, 2000; see also Rapini,
2002, and references therein), which is here supported by
molecular data.
Asclepiadinae sensu Liede (1997b) are confirmed,
perhaps with Pergularia as sister to the rest (Fig. 5) and
Asclepias nested within it. Metastelmatinae sensu Liede
(1997b), on the other hand, are shown to be paraphyletic, and a division between members of New and Old
World was found (Liede & Täuber, 2000; Liede, 2001),
culminating in the re-instatement of Cynanchinae (Liede
& Täuber, in press). Our study does support the splitting
of Cynanchinae and Metastelmatinae, but the unresolved
position of Cynachum acutum (the type of Cynanchum)
in the ACTG clade prevents further consideration of their
circumscription (Fig. 5). American Cynanchum appear to
be more related to Metaplexis and the Glossonema clade
than to the other Cynanchum, forming a group characterised by terminals with long branches (Fig. 5).
Cynanchinae, as circumscribed by Liede & Täuber
(2002), were not detected, which may be caused by the
39
Rapini & al. Asclepiadeae classification
52 February 2003: 33–50
100
96/92
100
86/86
99
63/63
Gonolobinae
Metastelmatinae
Oxypetalinae
95
--/--
95
--/-100
--/--
100
--/55
100
98/99
97
80/63
98
88/63
100
*/61
100
80/73
100
69/66
100
--/ *
100
71/71
100
74/75
Schubertia grandiflora
Gonolobus rostratus
Gonolobus selloanus
Matelea pedalis
Schystogyne sylvestris
Oxypetalum banksii
Oxypetalum appendiculatum
Oxypetalum coeruleum
Oxypetalum capitatum
Oxypetalum sublanatum
Oxypetalum minarum
Oxypetalum insigne
Oxypetalum strictum
Araujia sericifera
Melinia parviflora
Melinia discolor
Melinia candolleana
Philibertia vailiae
Philibertia lysimachioides
Funastrum clausum
Funastrum odoratum
Tassadia berteriana
Tassadia obovata
Metastelma parviflorum
Petalostelma sarcostemma
Metastelma linearifolium
Metastelma schaffneri
Blepharodon nitidum
Blepharodon mucronatum
Blepharodon lineare
Nautonia nummularia
Hemipogon acerosus
Hemipogon luteus
Hemipogon carassensis
Ditassa cordeiroana
Ditassa hastata
Ditassa banksii
Ditassa acerosa
Ditassa decussata
Ditassa grazielae
Ditassa magisteriana
Ditassa ditassoides
Ditassa tomentosa
Gonioanthela hilariana
Nephradenia asparagoides
Nephradenia acerosa
Barjonia chloraeifolia
Jobinia lindbergii
Metastelma scoparium
Orthosia urceolata
Ditassa subtrivialis
Cynanchum morrenioides
Cynanchum nummulariifolium
Astephanus geminiflorus
Grisebachiella hieronymi
Fig. 4. Consensus tree (posterior probability >94%) of Asclepiadoideae resulting from 40,000 trees generated from
Bayesian analysis of trnL-F. The tribal (black bar, Marsdenieae; stippled bar, Ceropegieae; white bar, Asclepiadeae) and
subtribal classification of Asclepiadoideae with emphasis on Asclepiadeae subtribes sensu Liede (1997b; depicted in
Fig. 2) is mapped onto this topology. Numbers above branches indicate percentage of posterior probability for the
clade. Numbers bellow branches indicate bootstrap percentage >49% from analyses of matrix 1 (considering gap information) and matrix 2 (not considering gap information), respectively. * = clades not present in the strict consensus tree;
-- = clades <50% bootstrap present in the strict consensus tree. Upper portion of the consensus tree.
40
Rapini & al. Asclepiadeae classification
52 February 2003: 33–50
Asclepiadinae
Astephaninae
Glossonematinae
Kanahia laniflora
100
78/86
Calotropis procera
Stathmostelma gigantiflorum
Metastelmatinae
Stenostelma corniculatum
Schizoglossum alpestre
99
72/61
98
72/53
Pachycarpus spurius
99
68/63
Margaretta rosea
Gomphocarpus fruticosus
Aspidoglossum ovalifolium
100
94/78
Glossostelma spathulatum
Xysmalobium undulatum
98
66/64
100
87/85
Asclepias syriaca
Asclepias curassavica
Asclepias mellodora
Pergularia daemia
Solenostemma arghel
100
100/100
100
87/87
Glossonema boveanum
Odontanthera radians
Pentarrhinum insipidum
Metaplexis japonica
Oxystelma esculentum
100
--/ *
Karimbolea verrucosa
99
61/54
Folotsia grandiflora
100
68/69
Platykeleba insignis
Sarcostemma viminale
Schizostephanus alatus
Cynanchum acutum
Cynanchum obovatum
98
82/70
100
78/78
Cynanchum montevidense
Cynanchum albiflorum
Cynanchum laeve
Biondia henryi
100
88/85
100
74/70
Cynanchum roulinioides
100
99/99
100
83/79
Blyttia fruticulosa
100
64/65
Diplostigma canescens
Goydera somaliensis
Pentatropis nivalis
Vincetoxicum hirundinaria
Tylophora flexuosa
Microloma tenuifolium
100
100/100
Oncinema lineare
Astephanus triflorus
Eustegia minuta
Cionura erecta
Dregea sinensis
98
50/ *
98
82/72
100
71/70
99
65/66
Marsdenia suberosa
98
62/60
Marsdenia zehntneri
Marsdenia amorimii
Telosma cordata
100
88/75
Micholitzia obcordata
100
99/98
100
100/100
Hoya australis
Dischidia bengalensis
100
100/100
Stapelia leendertziae
Stapelia glanduliflora
Ceropegia saxatilis
Fockea edulis
Periploca graeca
Apocynum androsaemifolium
Secamone glaberrima
Fig. 4 (continued). Lower portion of Asclepiadoideae consensus tree.
41
Rapini & al. Asclepiadeae classification
52 February 2003: 33–50
Oxypetalum coeruleum
2
Melinia parviflora
1
Melinia discolor
1 1
Melinia candolleana
94
Philibertia vailiae
1
1
*
1
Philibertia lysimachioides
3
1
*1
*
Araujia sericifera
Schystogyne sylvestris
2
Oxypetalinae
Oxypetalum banksii
Oxypetalum appendiculatum
2
Oxypetalum sublanatum
Oxypetalum capitatum
1
2
Oxypetalum minarum
Oxypetalum insigne
1
Oxypetalum strictum
2
2
*3
1
Blepharodon nitidum
Blepharodon mucronatum
Metastelma parviflorum
1
Metastelma schaffneri
6
Metastelma linearifolium
Nephradenia acerosa
2
2
1
Core
Group
Nephradenia asparagoides
Gonioanthela hilariana
3
Ditassa hastata
1 Hemipogon acerosus
2
Petalostelma sarcostemma
1
93/65
2
*
3
2
78
2
1
*1
Ditassa cordeiroana
Nautonia nummularia
1
Hemipogon carassensis
Ditassa banksii
1
6
Ditassa acerosa
Ditassa grazielae
1
*
Metastelmatinae
Barjonia chloraeifolia
Hemipogon luteus
2
*
Ditassa magisteriana
Ditassa ditassoides
2
Ditassa decussata
Ditassa tomentosa
5
MOG
2
2
2
*
1
4
*
Funastrum odoratum
4
3
2
*
*
Tassadia berteriana
Cynanchum nummulariifolium
Grisebachiella hieronymi
3
Ditassa subtrivialis
7
2
Cynanchum morrenioides
4
2
*
Matelea pedalis
Tassadia obovata
Astephanus geminiflorus
7
1
*
Gonolobinae
1
4
64
Gonolobus rostratus
Gonolobus selloanus
2
*5
1
Schubertia grandiflora
2
3
5
1
Blepharodon lineare
Funastrum clausum
2
*
2
3
Jobinia lindbergii
Metastelma scoparium
Orthosia urceolata
5 changes
Fig. 5. Majority-rule consensus tree of 40,000 trees generated from Bayesian analysis of trnL-F, showing an updated
classification after phylogenetic studies based on molecular data: Asclepiadinae sensu Liede (1997b), Astephaninae
and Tylophorinae sensu Liede (2001), and Cynanchinae sensu Liede & Täuber (2002). Numbers of substitutions are
indicated above branches. Percentage of posterior probabilities of clades >50% and <95% are under branches, with
bootstrap percentage >50% from analysis of matrix 1 (containing gap information). * = clades >94% probability.
42
Rapini & al. Asclepiadeae classification
52 February 2003: 33–50
2
MOG
*
1 Karimbolea verrucosa
1
Folotsia grandiflora
Sarcostemma viminale
2
*
2
1
2
*
2
*
2
1
Cynanchinae
Platykeleba insignis
2
Kanahia laniflora
2
5
Calotropis procera
*
Stathmostelma gigantiflorum
2
Stenostelma corniculatum
2
Pachycarpus spurius
1
Xysmalobium undulatum
3
Schizoglossum alpestre
*
Glossostelma spathulatum
1
Margaretta rosea
1
Asclepiadinae
6
7
1
Gomphocarpus fruticosus
Aspidoglossum ovalifolium
Asclepias syriaca
3
1
Asclepias curassavica
2
2
*
Asclepias mellodora
* Pergularia
7
daemia
1 Schizostephanus alatus
Cynanchinae
1
Blyttia fruticulosa
2
Diplostigma canescens
* 1 1 Goydera somaliensis
*
Tylophorinae
1 Tylophora flexuosa
3
* 3 Vincetoxicum hirundinaria
Biondia henryi
6
Pentatropis nivalis
*
93/84
6
1
2
1
*
21
1
*
*
*
11
*
4
1
*
1 1
*
8
3
*
*
2
*
14
4
Fockea edulis
Cynanchum albiflorum
Cynanchum laeve
Cynanchinae
4
2
4
*
4
86/52
*
Pentarrhinum insipidum
4
7
Glossonema boveanum
9
*
1 Odontanthera radians
*
18
Metaplexis japonica
Solenostemma arghel
15
Oxystelma esculentum
Microloma tenuifolium
2 Astephanus triflorus
Astephaninae
2
Oncinema lineare
4
Cionura erecta
7
Telosma cordata
6
3
10
8
2
67/86
5
Cynanchum acutum
1 Cynanchum obovatum
57
6
Cynanchum roulinioides
2
12
Cynanchum montevidense
1 60
8
5
17
4
*
ACTG
Dregea sinensis
Marsdenia suberosa
Marsdenia amorimii
Marsdenia zehntneri
Marsdenieae
6
Micholitzia obcordata
Hoya australis
6
Dischidia bengalensis
2
Stapelia leendertziae
6
2
Stapelia glandulifora
24
2 *
Ceropegia saxatilis
*
Eustegia minuta
Eustegiinae
3
2
Ceropegieae
21
Periploca graeca
Apocynum androsaemifolium
Secamone glaberrima
12
5
5 changes
Fig. 5 (continued). Lower portion of Asclepiadoideae of the majority-rule consensus tree. ACTG-clade including
Asclepiadinae, Cynanchinae, Tylophorinae and Glossonematinae. MOG-clade including Metastelmatinae, Oxypetalinae
and Gonolobinae. Black bar, taxa with erect pollinia; white bar, taxa with pendent pollinia. New World clades are indicated by bold lines.
43
Rapini & al. Asclepiadeae classification
52 February 2003: 33–50
Table 2. Classification of genera treated in this study according to Schumann (1895), more recent general classifications (Fig. 2), and phylogenetic studies summarised in this paper (Figs. 4 and 5). Tribes are in all capital letters; groups
are recognised in Asclepiadeae, unless mentioned. ACTG –group comprises members of Asclepiadinae, Cynanchinae,
Tylophorinae, and Glossonematinae; MOG – group comprises members of Metastelmatinae, Oxypetalinae and
Gonolobinae (Fig. 5). * denotes circumscription that contradicts the main patterns of relationship obtained with trnL-F.
Liede & Albers, 1994;
Phylogenetics
Genus
Schumann, 1895
Liede, 1997b (Fig. 2)
(Figs. 4 and 5)
Araujia Brot.
Glossonematinae*
Oxypetalinae
Oxypetalinae
Asclepias L.
Asclepiadinae
Asclepiadinae
Asclepiadinae
Aspidoglossum E. Mey.
Asclepiadinae
Asclepiadinae
Asclepiadinae
Astephanus R. Br.
Astephaninae
Astephaninae
Astephaninae
Barjonia Decne.
TYLOPHOREAE (Marsdeniinae)*
MARSDENIEAE*
ASCLEPIADEAE (Metastelmatinae)
Biondia Schltr.
1905
Metastelmatinae*
Tylophorinae
Blepharodon Decne.
Asclepiadinae*
Metastelmatinae
Metastelmatinae
Blyttia Arn.
Cynanchinae*
Astephanineae*
Tylophorinae
Calotropis R. Br.
Asclepiadinae
Asclepiadinae
Asclepiadinae
Ceropegia L.
TYLOPHOREAE (Ceropegiinae)
CEROPEGIEAE
CEROPEGIEAE
Cionura Griseb.
TYLOPHOREAE (Marsdeniinae)
MARSDENIEAE
MARSDENIEAE
Cynanchum L.
Cynanchinae
Metastelmatinae*
Cynanchinae
Diplostigma K.Schum.
Astephaninae*
Astephaninae*
ACTG
Dischidia R. Br.
TYLOPHOREAE (Marsdeniinae)
MARSDENIEAE
MARSDENIEAE
Ditassa R. Br.
Asclepiadinae*
Metastelmatinae
Metastelmatinae
Dregea E. Mey.
TYLOPHOREAE (Marsdeniinae)
MARSDENIEAE
MARSDENIEAE
Eustegia R. Br.
Asclepiadinae*
Astephanineae*
Eustegiinae
Fockea Endl.
TYLOPHOREAE (Marsdeniiinae)*
FOCKEEAE
FOCKEEAE
Folotsia Costantin & Bois
1908
Metastelmatinae*
ACTG
Funastrum E. Fourn.
Asclepiadinae*
Metastelmatinae
MOG
Glossonema Decne.
Glossonematinae
Glossonematinae
ACTG
Glossostelma Schltr.
1895
Asclepiadinae
Asclepiadinae
Gomphocarpus R. Br.
Asclepiadinae
Asclepiadinae
Asclepiadinae
Gonioanthela Malme
1927
Metastelmatinae
Metastelmatinae
Gonolobus Michx.
GONOLOBEAE*
ASCLEPIADEAE (Gonolobinae)
ASCLEPIADEAE (Gonolobinae)
Goydera Liede
1993
Astephaninae*
ACTG
Grisebachiella Lorentz
?
Metastelmatinae*
MOG (but outside core group)
Hemipogon Decne.
Astephaninae*
Metastelmatinae
Metastelmatinae
Hoya R. Br.
TYLOPHOREAE (Marsdeniinae)
MARSDENIEAE
MARSDENIEAE
Jobinia E. Fourn.
TYLOPHOREAE (Marsdeniinae)*
MARSDENIEAE*
ASCLEPIADEAE (MOG)
Kanahia R. Br.
Asclepiadinae
Asclepiadinae
Asclepiadinae
Karimbolea Descoings
1960
Metastelmatinae*
ACTG
Margaretta Oliv.
Asclepiadinae
Asclepiadinae
Asclepiadinae
Marsdenia R. Br.
TYLOPHOREAE (Marsdeniinae)
MARSDENIEAE
MARSDENIEAE
Matelea Aubl.
GONOLOBEAE*
ASCLEPIADEAE (Gonolobinae)
ASCLEPIADEAE (Gonolobinae)
Melinia Decne.
Asclepiadinae*
Oxypetalinae
Oxypetalinae
Metaplexis R. Br.
Cynanchinae
Metastelmatinae*
ACTG
Metastelma R. Br.
Asclepiadinae*
Metastelmatinae
Metastelmatinae
Micholitzia N. E.Br.
1909
MARSDENIEAE
MARSDENIEAE
Microloma R. Br.
Astephaninae
Astephaninae
Astephaninae
Nautonia Decne.
Astephaninae*
Metastelmatinae
Metastelmatinae
Nephradenia Decne.
TYLOPHOREAE* (Marsdeniinae)
MARSDENIEAE*
ASCLEPIADEAE (Metastelmatinae)
OdontantheraWight
Glossonematinae
Glossonematinae
ACTG
Oncinema Arn.
Cynanchinae?*
Astephaninae
Astephaninae
Orthosia Decne.
Cynanchinae*
Metastelmatinae*
MOG (but outside core group)
Oxypetalum R. Br.
Oxypetalinae
Oxypetalinae
Oxypetalinae
Oxystelma R. Br.
Glossonematinae
Metastelmatinae*
ACTG
Pachycarpus E. Mey.
Asclepiadinae
Asclepiadinae
Asclepiadinae
44
52 February 2003: 33–50
Rapini & al. Asclepiadeae classification
Table 2. (continued).
Liede & Albers, 1994;
Phylogenetics
Genus
Schumann, 1895
Liede, 1997b (Fig. 2)
(Figs. 4 and 5)
Pentarrhynum E. Mey.
Asclepiadinae
Metastelmatinae*
ACTG
Pentatropis R. Br.
Cynanchinae
Astephaninae*
ACTG
Pergularia L.
TYLOPHOREAE* (Marsdeniinae) ASCLEPIADEAE (Asclepiadinae) ASCLEPIADEAE (Asclepiadinae)
Petalostelma E. Fourn. TYLOPHOREAE* (Marsdeniinae)ASCLEPIADEAE (Metastelmatinae) ASCLEPIADEAE (Metastelmatinae)
Philibertia Kunth.
Glossonematinae*
Metastelmatinae*
Oxypetalinae
Platykeleba N.E. Br.
1895
Metastelmatinae*
ACTG
Sarcostemma R. Br.
Cynanchinae
Metastelmatinae*
ACTG
Schistogyne Hook. & Arn.
Asclepiadinae*
Oxypetalinae
Oxypetalinae
SchizoglossumE.Mey.
Asclepiadinae
Asclepiadinae
ACTG
SchizostephanusHochst. ex Benth. Cynanchinae
Astephaninae*
ACTG
Schubertia Mart.
Glossonematinae*
Gonolobinae
Gonolobinae
Solenostemma Hayne
Glossonematinae
Glossonematinae
ACTG
Stapelia L.
TYLOPHOREAE (Ceropegiinae)
CEROPEGIEAE
CEROPEGIEAE
Stenostelma Schltr.
Asclepiadinae
Asclepiadinae
Asclepiadinae
Stathmostelma K. Schum.
Asclepiadinae
Asclepiadinae
Asclepiadinae
Tassadia Decne.
Asclepiadinae*
Metastelmatinae
MOG
Telosma Coville
1905
MARSDENIEAE
MARSDENIEAE
Tylophora R. Br.
TYLOPHOREAE (Marsdeniinae)* ASCLEPIADEAE (Astephaninae)*
ASCLEPIADEAE (ACTG)
Vincetoxicum Wolf.
Cynanchinae
Astephaninae*
ACTG
Xysmalobium R. Br.
Asclepiadinae
Asclepiadinae
Asclepiadinae
smaller sequence as well as the inclusion of a greater
diversity of ACTG taxa used in our study. According to
their study, Cynanchinae would comprise Cynanchum
and Sarcostemma, as well as the small genera
Schizostephanus, Glossonema, Metaplexis, Pentarrhinum, Odontanthera, Folotsia, Karimbolea and
Platykeleba. Cynanchum has had an unstable circumscription, and opinions on its delimitation have varied
drastically. Nevertheless, Woodson’s (1941) concept of
Cynanchum including many American genera of
Metastelmatinae as well as Liede’s (1997c) infrageneric
classification of the American Cynanchum may be discarded (see Liede & Täuber, 2002, for details on
Cynanchum classification). The re-classification of several species of American Cynanchum (e.g., C. morrenioides and C. nummulariifolium) that are clearly unrelated to the C. acutum clade is required; others (e.g., C.
montevidense and C. roulinioides; Fig. 5) might belong
to Cynanchum (Liede & Täuber, 2002). These conclusions, however, are derived from the MOG–ACTG
dichotomy (Fig. 5); the appropriate circumscription for
Cynanchum and even Cynanchinae among ACTG groups
is still unclear (Fig. 5).
MOG represents the most diverse New World group,
ranging from the showy, apocynoid-like flowers of
Schubertia to the tiny flowers of Tassadia and from the
minute sclerophyllous leaves of erect subshrubs of some
Ditassa to the large leaves of long vines of Gonolobus.
This huge morphological diversity, however, is not
reflected in molecular diversity of trnL-F, contrasting
with the morphologically less diverse American
Cynanchum clade, which is characterised by a much
greater level of molecular change. The three subtribes of
MOG (Metastelmatinae, Oxypetalinae and Gonolobinae)
can be recognised in the DNA tree, but the limited numbers of substitutions within the clades are insufficient to
delineate generic patterns. Metastelmatinae, even
reduced to American groups, are still unsatisfactory, and
the exclusion of Orthosia and Jobinia as well as the
transfer of Philibertia to Oxypetalinae should be considered. The position of the Orthosia-Jobinia clade and
other species outside the MOG core group (Fig. 3b) indicates the need for new subtribes to accommodate these
New World groups. Gonolobinae genera form a wellsupported group but of uncertain position in the MOG
clade. Their inclusion in Asclepiadeae is appropriate,
whereas their reinstatement at tribal level suggested by
Endress & Stevens (2001) would be hierarchically
incongruent.
CONCLUSIONS
These data, analysed by both Bayesian inference and
parsimony methods, detected major patterns of relationships in Asclepiadoideae. Excluding the lack of evidence
for Cynanchinae, the topology is congruent with those
presented in studies that also included the trnT-L intergenic spacer, and confidence in clades estimated by bootstrap and posterior probability provides an evaluation of
45
Rapini & al. Asclepiadeae classification
current knowledge of phylogenetic relationships in
Asclepiadeae. Since Eustegia might be sister to the
Marsdenieae–Ceropegieae clade and treated as the only
genus of the very reduced Eustegiinae Decne.,
Astephaninae may represent the sister group of the other
Asclepiadeae. The rest of the tribe can then be divided
into ACTG, including two American groups, and MOG,
restricted to the New World. In the ACTG clade,
Asclepiadinae and Tylophorinae are supported, but this is
not true for Cynanchinae and Glossonematinae.
Relationships among components of the ACTG clade are
not clear, and the unresolved position of the nomenclaturally critical species Cynanchum acutum in a weakly supported clade of Cynanchinae permits several alternative hypotheses. Taxonomic changes (e.g., Liede &
Meve, 2001) based exclusively on these molecular data,
therefore, are premature. Conclusions about Glossonematinae (Liede & al., 2002) are also tied to the resolution of ACTG components and the consequent definition
of Cynanchinae.
Although not linked to any published phylogenetic
analysis, Liede’s (1997b) arrangement of genera is closer to the one coming out of phylogenetic studies than is
Schumann’s (1895) classification (Table 2). In only five
years, however, several changes have been made, and
many others are still needed to achieve a classification
based on monophyletic groups. The phylogeny of New
World Asclepiadoideae has not yet been well explored,
and, until a better resolution of relationships indicates a
reasonable generic rearrangement, taxonomic revisions
of such groups are likely to produce ephemeral publications. Improvement in the understanding of Asclepiadoideae is evident, but it is too early to formalise taxonomic groups based on such weak patterns as those
observed in our analyses of trnL-F.
In this overview, four clades of New World
Asclepiadoideae were detected (Fig. 3). (1) The
American Marsdenia (Marsdenieae), comprising around
60 species, may form a monophyletic group, but analyses including a large diversity of the subtribe are needed
to confirm this. (2) American Cynanchum seems to be
restricted to the subgenus Mellichampia (Liede &
Täuber, 2002), which may include C. roulinioides and
the species classified in Metalepis Griseb., comprising a
group with around 20 species. However, according to the
molecular data presented here, this group seems to be
more closely related to the Glossonema clade and
Metaplexis than to other Cynanchum. (3) Asclepias with
around 130 species, most of which are distributed in
North and Central America, represents the only
American group of Asclepiadinae and is nested within
African genera. (4) MOG comprises members of the
exclusively American Metastelmatinae (including
species of “Cynanchum” and “Astephanus” that do not
46
52 February 2003: 33–50
belong to the ACTG clade), Oxypetalinae, and
Gonolobinae. With around 600 species, the MOG clade
includes the great majority of American
Asclepiadoideae, having their centre of diversity in
South America.
Based on these results, three main topics should be
explored for an improved understanding of New World
Asclepiadoideae: (1) the relationship between American
Marsdenia and the rest of Marsdenieae, especially the
Old World Marsdenia groups; (2) the position of
Cynanchum subgenus Mellichampia in the ACTG clade
for which the placement of C. acutum is crucial; and (3)
a massive study of relationships within the large and
morphologically diverse MOG clade.
ACKNOWLEDGEMENTS
This paper is part of the results of a postdoctoral fellowship
of the first author (AR) at Royal Botanic Gardens, Kew, funded by
the Mellon Foundation. Most of the material sequenced for this
study was collected during the Ph.D. of AR, funded by Fundação
de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and
Conselho de Aperfeiçoamento de Pessoal de Ensino Superior
(CAPES). Renato de Mello-Silva, Rafaela C. Forzza, Tatiana
Konno, and Maria Ana Farinaccio kindly provided material for
DNA extraction, and Sigrid Liede gave access to unpublished
manuscripts (and sequences used therein). We also thank Mary
Endress for reading and encouraging this publication and an
anonymous reviewer for criticisms and suggestions.
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Appendix 1. List of taxa with distributions and GenBank (www.ncbi.nlm.nih.gov) accession numbers. Voucher information is provided for samples sequenced in this study (AY163662–AY163725; herbarium acronyms according to
Holmgren & al., 1990). Sequences not yet available on GenBank were obtained from Liede’s homepage (www.unibayreuth.de/departments/planta2/wgl/fsigrid2.html). i. s. = intergenic spacer; NW = New World; OW = Old World; ? = cultivated source and/or without stated provenance.
Species
Distribution
Apocynum androsaemifolium L.
U.S.A.
Araujia sericifera Brot.
Australia (native to NW)
Asclepias curassavica L.
Brazil
A. mellodora A.St.-Hil.
Brazil
A. syriaca L.
Germany (native to NW)
Aspidoglossum ovalifolium (Schltr.) Kupicha
South Africa
Astephanus geminiflorus Decne.
Chile
A. triflorus R. Br.
South Africa
Barjonia chloraeifolia Decne.
Brazil
Biondia henryi (Warb. ex Schltr. & Diels) Tsiang & P.T. Li China
Blepharodon lineare (Decne.) Decne.
Argentina
B. mucronatum (Schltdl.) Decne.
Belize
B. nitidum (Vell.) J.F. Macbr.
Brazil
Blyttia fruticulosa (Decne.) D.V. Field
Kenya
Calotropis procera (Aiton) W.T. Aiton
Puerto Rico (native to OW)
Ceropegia saxatilis Jum. & H. Perrier
Madagascar
Cionura erecta Griseb.
Turkey
Cynanchum acutum L.
Egypt
C. albiflorum (Urban) Liede
Ecuador
C. laeve (Michx.) Pers.
U.S.A.
C. montevidense Spreng.
Argentina
C. morrenioides Goyder
Brazil
C. nummulariifolium Hook. & Arn.
Argentina
C. obovatum Choux.
Madagascar
C. roulinioides (E. Fourn.) Rapini
Brazil
Diplostigma canescens K. Schum.
Kenya
Dischidia bengalensis Colebr.
? (OW)
Ditassa acerosa Mart.
Brazil
D. banksii R.Br. ex Schult.
Brazil
D. cordeiroana Fontella
Brazil
D. decussata Mart.
Brazil
D. ditassoides (Silveira) Fontella
Brazil
D. grazielae Fontella & Marquete
Brazil
D. hastata Decne.
Brazil
D. magisteriana Rapini
Brazil
D. subtrivialis Griseb.
Bolivia
D. tomentosa (Vell.) Fontella
Brazil
48
trnL intron
trnL-F i. s.
AF214308
AY163662
AY163664
AY163665
AF214311
AY163666
AJ410182
AJ410188
AY163667
AJ410191
AY163668
AJ290840
AY163669
AJ410194
AF214324
AJ410041
AJ410173
AY163670
AJ428775
AJ428652
AJ290849
AJ428685
AJ290852
AJ428802
AY163672
AJ410200
AF214343
AY163673
AY163674
AY163675
AY163677
AY163678
AY163679
AY163680
AY163681
AJ428755
AY163682
AF214154
AY163663
AY163664
AY163665
AJ410180
AY163666
AJ410183
AJ410189
AY163667
AJ410192
AY163668
AJ290841
AY163669
AJ410195
AF214170
AJ410042
AJ410174
AY163671
AJ428776
AJ428653
AJ290850
AJ428684
AJ290851
AJ428803
AY163672
AJ410201
AF214189
AY163673
AY163674
AY163676
AY163677
AY163678
AY163679
AY163680
AY163681
AJ428756
AY163682
Voucher
Forster 7656 (K)
Rapini 933 (SPF)
Rapini 411 (SPF)
Balkwill 10847 (K)
Rapini 485 (SPF)
Forzza 2027 (SPF)
Rapini 938 (SPF)
Boulos s.n. (K)
Rapini 726 (SPF)
Rapini 383 (SPF)
Konno 754 (SPF)
Rapini 347 (SPF)
Rapini 556 (SPF)
Rapini 557 (SPF)
Rapini 379 (SPF)
Rapini 777 (SPF)
Rapini 597 (SPF)
Rapini 616 (SPF)
Rapini & al. Asclepiadeae classification
52 February 2003: 33–50
Appendix 1 (continued).
Species
Distribution
Dregea sinensis Hemsl.
? (OW)
Eustegia minuta (L.f.) N.E. Br.
South Africa
Fockea edulis K.Schum.
? (OW)
Folotsia grandiflora (Jum. & H. Perrier)
Madagascar
Jum. & H. Perrier
Funastrum clausum (Jacq.) Schltr.
Argentina
F. odoratum (Hemsl.) Schltr.
Mexico
Glossonema boveanum (Decne.) Decne.
Yemen
Glossostelma spathulatum (K. Schum.) Bullock
Zimbabwe
Gomphocarpus fruticosus (L.) W.T. Aiton
Egypt
Gonioanthela hilariana (E. Fourn.) Malme
Brazil
Gonolobus rostratus (Vahl) Schult.
Ghana? (native NW)
G. selloanus (E. Fourn.) Bacigalupo
Brazil
Goydera somaliensis Liede
Somalia
Grisebachiella hieronymi Lorentz
Argentina
Gymnema sylvestre (Retz.) R. Br. ex Schult.
Cameroon
Hemipogon acerosus Decne.
Brazil
H. carassensis (Malme) Rapini
Brazil
H. luteus E.Fourn.
Brazil
Hoya australis R. Br. ex J. Traill
? (OW)
Jobinia lindbergii E. Fourn.
Brazil
Kanahia laniflora (Forssk.) R. Br.
Tanzania
Karimbolea verrucosa Desc.
Madagascar
Margaretta rosea Oliv.
Tanzania
Marsdenia amorimii Morillo
Brazil
M. suberosa (E. Fourn.) Malme
Brazil
M. zehntneri Fontella
Brazil
Matelea pedalis (E. Fourn.) Fontella & E.A. Schwarz
Brazil
Melinia candolleana (Hook. & Arn.) Decne.
Argentina
M. discolor Schltr.
Argentina
M. parviflora (Malme) Krap. & Cáceres
Argentina
Metaplexis japonica Makino
Russia
Metastelma linearifoliumA. Rich
? (NW)
M. parviflorum (Sw.) R. Br. ex Schult.
Puerto Rico
M. schaffneri A. Gray
Mexico
M. scoparium (Nutt.) Vail
Brazil
Micholitzia obcordata N.E. Br.
Thailand
Microloma tenuifolium K. Schum.
South Africa
Nautonia nummularia Decne.
Argentina
Nephradenia acerosa Decne.
Brazil
N. asparagoides (Decne.) E. Fourn.
Brazil
Odontanthera radians (Forssk.) D.V. Field
Yemen
Oncinema lineare (L.f.) Bullock
South Africa
Orthosia urceolata E. Fourn.
Brazil
Oxypetalum appendiculatumMart.
Brazil
O. banksii R. Br. ex Schult.
Brazil
O. capitatum Mart.
Argentina
Oxypetalum coeruleum (D. Don ex Sweet) Decne.
? (NW)
O. insigne (Decne.) Malme
Brazil
O. minarum E. Fourn.
Brazil
O. strictum Mart.
Brazil
O. sublanatum Malme
Brazil
trnL intron
trnL-F i. s.
AF214345
AJ410206
AF214353
AJ290855
AF214191
AJ410207
AF214199
AJ290856
AY163683
AJ290873
AY163684
AY163686
AY163687
AY163688
AF214362
AY163689
AJ410209
AJ410212
AJ402137
AY163690
AY163692
AY163693
AF214367
AY163694
AY163695
AJ290880
AY163696
AF214377
AY163697
AY163698
AY163699
AJ410176
AY163700
AJ410224
AJ428811
AY163683
AJ290874
AY163685
AY163686
AY163687
AY163688
AF214208
AY163689
AJ410210
AJ410213
AJ402142
AY163691
AY163692
AY163693
AF214213
AY163694
AY163695
AJ290879
AY163696
AF214223
AY163697
AY163698
AY163699
AJ410177
AY163700
AJ410225
AJ428812
AY163701
AJ410215
AY163703
AF214381
AJ410221
AJ410227
AY163704
AY163706
AJ428814
AJ410230
AY163708
AY163709
AY163710
AY163711
AF214443
AY163712
AY163713
AY163714
AY163715
AY163702
AJ410216
AY163703
AF214227
AJ410222
AJ410228
AY163705
AY163707
AJ428815
AJ410231
AY163708
AY163709
AY163710
AY163711
AF214289
AY163712
AY163713
AY163714
AY163715
Voucher
Mello-Silva 1919 (SPF)
Rowaished 3014 (K)
Goyder 4108 (K)
Chase 9370 (K)
Rapini 710 (SPF)
Rapini 609 (SPF)
Forzza 599 (SPF)
Rapini 778 (SPF)
Forzza 610 (SPF)
Farinaccio 194 (SPF)
Goyder 3791 (K)
Goyder 3931 (K)
Rapini 384 (SPF)
Pirani 4399 (SPF)
Rapini 714 (SPF)
Mello-Silva 1887 (SPF)
Liede homepage
Axelrod 8328 (K)
Rapini 932 (SPF)
Philcox 3303 (K)
Irwin 13012 (K)
Rapini 934 (SPF)
Rapini 613 (SPF)
Rapini 911 (SPF)
Mello-Silva 1924 (SPF)
Rapini 716 (SPF)
Rapini 908 (SPF)
Rapini 811 (SPF)
Rapini 937 (SPF)
49
Rapini & al. Asclepiadeae classification
52 February 2003: 33–50
Appendix 1 (continued).
Species
Distribution
trnL intron
trnL-F i. s.
Oxystelma esculentum (L.f.) Sm.
Egypt
Pachycarpus spurius (N.E. Br.) Bullock
Tanzania
Pentarrhinum insipidum E.Mey
South Africa
Pentatropis nivalis (J.F. Gmel.) D.V. Field & J.R.I. Wood Kenya
Pergularia daemia (Forssk.) Chiov.
Tanzania
Periploca graeca L.
? (OW)
Petalostelma sarcostemma (Lillo) Liede & Meve
Argentina
Philibertia lysimachioides (Wedd.) T. Mey.
Argentina
P. vailiae (Rusby) Liede
Argentina
Platykeleba insignis N.E.Br.
Madagascar
Sarcostemma viminale (L.) R. Br.
Zimbabwe
Schistogyne sylvestris Hook. & Arn.
Argentina
Schizoglossum alpestre K. Schum.
Tanzania
Schizostephanus alatus Hochst. ex K. Schum.
Tanzania
Schubertia grandiflora Mart.
Argentina
Secamone glaberrima K. Schum.
Madagascar
Solenostemma arghel (Delile) Hayne
Egypt
Stapelia glanduliflora Masson
South Africa
S. leendertziae N.E. Br.
? (OW)
Stathmostelma gigantiflorum K.Schum.
Kenya
Stenostelma corniculatum (E. Mey.) Bullock
South Africa
Tassadia berteriana (Spreng.) W.D. Stevens
Bolivia
T. obovata Decne.
Costa Rica
Telosma cordata (Burm.f.) Merr.
? (OW)
Tylophora flexuosa R. Br.
Philippines
Vincetoxicum hirundinaria Medic.
Germany
Xysmalobium undulatum (L.) W.T. Aiton
South Africa
AJ290885
AY163716
AJ410233
AJ410239
AJ290892
AF102468
AJ 428787
AY163717
AJ290904
AJ290907
AJ290913
AJ410245
AY163718
AJ410248
AJ428826
AF214420
AY163719
AJ402128
AF214424
AY163721
AY163722
AJ428790
AY163723
AF214280
AJ290916
AJ410275
AY163725
AJ290887
AY163716
AJ410234
AJ410240
AJ290893
AF214244
AJ428788
AY163717
AJ290905
AJ290906
AJ290912
AJ10246
AY163718
AJ410249
AJ428827
AF214266
AY163720
AJ402151
AF214270
AY163721
AY163722
AJ428791
AY163724
AF102493
AJ290917
AJ410276
AY163725
50
Voucher
Goyder 3781 (K)
Mello-Silva 1886 (SPF)
Goyder 3892 (K)
Boulos19142 (K)
Harvey 64 (K)
Balkwill 10908 (K)
Angulo 300 (K)
Balkwill 10846 (K)