PHYLOGENY OF THE
HERBACEOUS TRIBE
SPERMACOCEAE (RUBIACEAE)
BASED ON PLASTID DNA DATA1
Inge Groeninckx,2 Steven Dessein,2,3
Helga Ochoterena,4 Claes Persson,5
Timothy J. Motley,6 Jesper Kårehed,7
Birgitta Bremer,7 Suzy Huysmans,2
and Erik Smets2,8
ABSTRACT
In its current circumscription, the herbaceous tribe Spermacoceae s.l. (Rubiaceae, Rubioideae) unites the former tribes
Spermacoceae s. str., Manettieae, and the Hedyotis–Oldenlandia group. Within Spermacoceae, and particularly within the
Hedyotis–Oldenlandia group, the generic delimitations are problematic. Up until now, molecular studies have focused on
specific taxonomic problems within the tribe. This study is the first to address phylogenetic relationships within Spermacoceae
from a tribal perspective. Sequences of three plastid markers (atpB-rbcL, rps16, and trnL-trnF) were analyzed separately as
well as combined using parsimony and Bayesian approaches. Our results support the expanded tribe Spermacoceae as
monophyletic. The former tribe Spermacoceae s. str. forms a monophyletic clade nested within the Hedyotis–Oldenlandia
group. Several genera formerly recognized within the Hedyotis–Oldenlandia group are supported as monophyletic (Amphiasma
Bremek., Arcytophyllum Willd. ex Schult. & Schult. f., Dentella J. R. Forst. & G. Forst., Kadua Cham. & Schltdl., and
Phylohydrax Puff), while others appear to be paraphyletic (e.g., Agathisanthemum Klotzsch), biphyletic (Kohautia Cham. &
Schltdl.), or polyphyletic (Hedyotis L. and Oldenlandia L. sensu Bremekamp). Morphological investigations of the taxa are
ongoing in order to find support for the many new clades and relationships detected. This study provides a phylogenetic
hypothesis with broad sampling across the major lineages of Spermacoceae that can be used to guide future species-level and
generic studies.
Key words: atpB-rbcL, Hedyotis–Oldenlandia group, Rubiaceae, molecular phylogeny, plastid DNA, rps16, Spermacoceae,
trnL-trnF.
The systematic relationships of the Rubiaceae
herbaceous representatives are still unclear at the
species and genus levels (Robbrecht & Manen,
2006). Even the higher-level classification in tribes
has been the subject of debate. In the last
comprehensive classification based on morphology
(Robbrecht, 1988, 1993), most herbaceous representatives were assigned to one of the following
tribes: Anthospermeae, Argostemmateae, Coccocypseleae, Hedyotideae, Knoxieae, Rubieae, Sipaneeae, Spermacoceae, and Theligoneae. Among
these, the Spermacoceae as traditionally delimited
(Hooker, 1873; Bremekamp, 1952, 1966; Verdcourt,
1958; Robbrecht, 1988, 1993), referred to in this
paper as Spermacoceae s. str., are characterized by
the presence of raphides, fimbriate stipules, uniovulate locules, seeds with an apparent adaxial
groove, and the frequent occurrence of pluriaperturate pollen grains. However, molecular data show
Spermacoceae s. str. to be deeply nested within the
Hedyotideae, making the latter tribe paraphyletic
(Bremer, 1996; Andersson & Rova, 1999; Bremer &
Manen, 2000; Dessein et al., 2005a). Therefore,
Bremer (1996) and later Bremer and Manen (2000)
proposed a wider definition for Spermacoceae, in
which the former tribes Spermacoceae s. str.,
Hedyotideae, Manettieae, Knoxieae, and Triainolepideae are merged.
1
We thank the organizers of the Third International Rubiaceae Conference and the editors of this volume for the invitation
to participate in the proceedings. We acknowledge the technical assistance of Anja Vandeperre from the Laboratory of Plant
Systematics, Katholieke Universiteit Leuven. Frank van Caekenberghe from the National Botanic Garden of Belgium is
thanked for providing lovely photographs of Spermacoceae. This research was supported by grants from the Fund for Scientific
Research–Flanders (FWO, G.0250.05 and G.0268.04). Inge Groeninckx holds a Ph.D. research grant from the Fund for
Scientific Research–Flanders.
2
Laboratory of Plant Systematics, Katholieke Universiteit Leuven, Kasteelpark Arenberg 31, P.O. Box 2437, BE-3001
Leuven, Belgium. Author for correspondence: inge.groeninckx@bio.kuleuven.be.
3
National Botanic Garden of Belgium, Domein van Bouchout, BE-1860 Meise, Belgium.
4
Instituto de Biologia, Universidad Nacional Autónoma de México, Apdo. Postal 70-367, 04510, Mexico.
5
Department of Plant and Environmental Sciences, Göteborg University, P.O. Box 461, SE-405 30 Göteborg, Sweden.
6
Department of Biological Sciences, Old Dominion University, 110 Mills Godwin Building, 45th Street, Norfolk, Virginia
23529-0266, U.S.A.
7
Bergius Foundation, Royal Swedish Academy of Sciences and Botany Department, Stockholm University, SE-106 91
Stockholm, Sweden.
8
National Herbarium of the Netherlands, Leiden University Branch, P.O. Box 9514, NL-2300 RA Leiden, The Netherlands.
doi: 10.3417/2006201
ANN. MISSOURI BOT. GARD. 96: 109–132. PUBLISHED ON 23 APRIL 2009.
110
Annals of the
Missouri Botanical Garden
Based on rps16 intron data, Andersson and Rova
(1999) also found that Hedyotideae is paraphyletic
relative to Spermacoceae s. str. They did not accept
the wide delimitation for Spermacoceae as proposed
by Bremer (1996), but suggested an emended tribe
Knoxieae that included a few genera of Hedyotideae
(i.e., Otiophora Zucc., Otomeria Benth., and Pentas
Benth.) as a more prudent taxonomic approach to
handle the information from molecular-based analyses. The latter view was followed by Dessein (2003),
who preferred to recognize an emended tribe Knoxieae
(including Knoxieae s. str., Triainolepideae, Otiophora, the Pentas group of Hedyotideae fide Dessein
et al. [2000], and Carphalea Juss.) as a sister group of
Spermacoceae (including Spermacoceae s. str., Manettieae, and most of Hedyotideae). Robbrecht and
Manen (2006), based on a supertree analysis of the
family, came to a similar conclusion and likewise
recognized Knoxieae s.l. and Spermacoceae s.l. The
monophyly of the former tribe has also been confirmed
by a subsequent molecular study by Kårehed and
Bremer (2007). In their taxonomic conspectus,
Robbrecht and Manen (2006) listed 33 genera of
Spermacoceae s.l. for which molecular sequence data
are available. Based on morphological data, we
recognize 31 of these 33 genera and consider that
the tribe should include 30 additional genera; these
are listed in Table 1. For each genus, the number of
species, the distribution, and the position in Robbrecht’s classification of 1988 are given.
Spermacoceae s.l. forms a primarily herbaceous
lineage that is generally characterized by fimbriate
stipules and 4-merous flowers. Floral characters
(Fig. 1), as well as seeds and fruits, are highly
variable. Morphologically, three main groups can be
identified within Spermacoceae s.l. The first, the
Hedyotis–Oldenlandia group, is characterized by
multiovulate locules and comprises the large genera
Hedyotis L. and Oldenlandia L. and their presumed
relatives. Most of these taxa were formerly placed in
the tribe Hedyotideae. The generic delimitations of
the Hedyotis–Oldenlandia group have been the
subject of controversy for many years. The main issue
is whether most species of the complex should be
lumped into Hedyotis (advocated by inter alia Merrill
& Metcalf, 1946; Wagner et al., 1989; Fosberg &
Sachet, 1991; Dutta & Deb, 2004) or whether many
small genera should be recognized in addition to a
narrow circumscription of Hedyotis and Oldenlandia
(supported for African taxa by Bremekamp, 1952; for
Neotropical taxa by Terrell et al., 1986; Terrell, 1991,
2001a, b, c; and for Asian taxa by Terrell & Robinson,
2003).
The second well-marked group within Spermacoceae s.l. is Spermacoceae s. str., which is character-
ized by uniovulate locules. According to Dessein
(2003), this group contains 19 genera of which
Spermacoce L. is by far the largest with an estimated
275 species. Within Spermacoceae s. str., controversy
has focused on the delimitation of its nominal genus
Spermacoce. The main question is whether Spermacoce
should be limited to species with the same type of fruit
dehiscence as S. tenuior L., the type species of the
genus. In this species, fruits open asymmetrically,
resulting in one closed and one open fruit part. If this
narrow delimitation for Spermacoce (referred to as
Spermacoce s. str.) is accepted, most other species in
the tribe Spermacoceae s. str. must be included in
Borreria G. Mey.
A third well-defined group within Spermacoceae
s.l. comprises only two American genera, Bouvardia
Salisb. and Manettia Mutis ex L. Bremekamp (1952)
considered Bouvardia closely related to Heterophyllaea Hook. f., Hindsia Benth. ex Lindl., and
Lecanosperma Rusby. Robbrecht (1988) placed these
genera together with inter alia Manettia in a group
with uncertain affinities, because their winged seeds
suggest a relation to Cinchoneae, while the presence
of raphides indicates a relation to Hedyotideae. In the
classification of Bremer and Manen (2000), only
Bouvardia and Manettia belong to Spermacoceae s.l.,
because Hindsia and Heterophyllaea (including Lecanosperma) are included in Coussareeae. Manettia is
similar to Bouvardia in many characters, but its
winding shoots and corneous endosperm separate it
from Bouvardia, which is erect and has fleshy
endosperm. These differences were the basis for
Bremekamp (1934) to place Manettia in its own tribe,
Manettieae.
Until now, molecular studies within Spermacoceae
s.l. have focused on particular taxonomic problems,
such as the circumscription and biogeography of
Arcytophyllum Willd. ex Schult. & Schult. f.
(Andersson et al., 2002), the generic status of
Houstonia L. (Church, 2003), the delimitation of
Pentanopsis Rendle, the affinities of Phylohydrax
Puff (Thulin & Bremer, 2004), and the taxonomic
position of Gomphocalyx Baker (Dessein et al.,
2005a). In the present paper, we aim to present a
phylogenetic hypothesis of Spermacoceae s.l. based
on the analysis of three plastid markers (atpB-rbcL,
rps16, and trnL-trnF) with the broadest sampling to
date. More specifically, we want to address the
following questions: (1) Is Spermacoceae s.l. as
circumscribed by Robbrecht and Manen (2006)
monophyletic? (2) What are the relationships among
members of Spermacoceae s. str. and genera of the
former tribes Hedyotideae and Manettieae? (3) What
are the major clades within the Hedyotis–Oldenlandia group?
Volume 96, Number 1
2009
Groeninckx et al.
Phylogeny of Spermacoceae
MATERIAL AND METHODS
1998). Sequences were initially aligned with ClustalX
(Thompson et al., 1997) applying the default parameters. Further adjustments of the preliminary aligned
data matrices were done manually with MacClade 4.04
(Maddison & Maddison, 2001). Parsimonious informative gaps were coded manually according to the
conservative simple indel coding method described
by Simmons and Ochoterena (2000).
PLANT MATERIAL AND SAMPLING
The aim was to obtain a broad sampling covering
most of the geographic and taxonomic diversity of
Spermacoceae and to enable identification of the
principal clades within the tribe. We included a total
of 128 species representing 32 of the 61 genera within
Spermacoceae. Three taxa belonging to the Knoxieae
(Batopedina pulvinellata Robbr., Carphalea madagascariensis Lam., and Pentanisia parviflora Stapf ex
Verdc.) were chosen as outgroup following Robbrecht
and Manen (2006) and Kårehed and Bremer (2007).
For rps16 and trnL-trnF, we used 40 and seven
previously published sequences, respectively (Andersson & Rova, 1999; Andersson et al., 2002; Dessein et
al., 2005a). Two hundred seventy-two sequences are
newly generated (100 atpB-rbcL sequences, 67 rps16
sequences, 105 trnL-trnF sequences) using dried silica
and herbarium material. Appendix 1 lists all taxa
included in this study with voucher information and
GenBank accession numbers.
DNA EXTRACTION, POLYMERASE CHAIN REACTION
AMPLIFICATION, AND SEQUENCING
DNA was extracted from silica-dried and herbarium
material using the CTAB method as described by
Janssens et al. (2006). Amplification of the atpB-rbcL
spacer was done with oligonucleotides two and five as
primers (Manen et al., 1994). Specific amplification
products could be obtained with a touchdown
polymerase chain reaction (PCR) with two cycles with
an annealing temperature of 53uC, then 12 cycles with
an annealing temperature of 52.5uC declining 0.5uC
every cycle, followed by 16 cycles with an annealing
temperature of 47uC. The rps16 intron was amplified
with the rps16F and rps16R2 primers described by
Oxelman et al. (1997). For the trnL-trnF intergenic
spacer, we used the primers e and f of Taberlet et al.
(1991). Both rps16 and trnL-trnF were amplified using
standard PCR techniques with an annealing temperature of 55uC. The PCR reaction mixture was cleaned
using a Nucleospin Extraction II Kit (Machery-Nagel,
Dren, Germany) according to the manufacturer’s
instructions. Sequencing was mostly done on an ABI
310 Genetic Analyzer (Applied Biosystems, Lennik,
Belgium). Some PCR products were sequenced by
Macrogen (Seoul, South Korea) sequencing facilities.
SEQUENCE ASSEMBLY, ALIGNMENT, AND GAP CODING
The assembling and editing of sequences were
conducted using the Staden Package (Staden et al.,
111
PHYLOGENETIC ANALYSES
Phylogenetic analyses were conducted using both
parsimony (MP) and Bayesian inference (BI). The
three plastid regions were first analyzed separately
and then combined.
Equally weighted MP analyses were performed
using Nona 2.0 (Goloboff, 1993) launched through
WinClada 1.00.08 (Nixon, 2002). Heuristic searches
for the shortest trees were performed using the
parsimony ratchet (Nixon, 1999). Ratchet runs of
200 iterations each, holding one tree per iteration and
randomly weighting 10% of the potentially informative characters, were carried out until the most
parsimonious trees (MPTs) were repeatedly found. A
strict consensus tree was calculated using the trees
obtained in the parsimony ratchet analyses. In order to
evaluate the relative support of the clades, jackknife
and bootstrap analyses were executed using 1000
replicates with 10 initial trees holding five trees per
random addition, doing tree bisection-reconnection
(TBR) to hold 1000 trees, and calculating a consensus
on each repetition. Frequency values were plotted
onto the consensus of the MPTs.
For the BI analyses, a substitution model was
selected for each DNA region with Modeltest 3.06
(Posada & Crandall, 1998) under the Akaike
Information Criterion (AIC). Modeltest selected the
GTR+I+G model of evolution for the atpB-rbcL spacer
and the GTR+G model for the two remaining markers.
Indels were not included in the BI analyses. In the
combined analysis, a mixed-model approach was used
(Ronquist & Huelsenbeck, 2003). The combined data
were partitioned and the same models of evolution
were used on the partitions as selected for the single
analyses. The BI analyses were conducted with
MrBayes 3b4 (Huelsenbeck & Ronquist, 2001). Four
Markov chains (one cold, three heated) starting with a
random tree were run simultaneously for one million
generations, sampling trees at every 100 generations.
The first 2500 sampled trees (25%) were regarded as
burn-in and discarded. PAUP* version 4b10 (Swofford, 2002) was used to calculate a 50% majority rule
tree and to report the posterior probabilities for each
clade. Only posterior probabilities above 0.95 are
considered (Suzuki et al., 2002).
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Annals of the
Missouri Botanical Garden
Table 1. List of genera associated with Spermacoceae s.l., their distribution, and species number following Govaerts et al.
(2006), except when stated otherwise. Genera in boldface were listed by Robbrecht and Manen (2006); other genera are here
based on morphological similarities. Synonymous taxa are as given by Robbrecht (1988), except when stated otherwise.
Genus
Robbrecht,
1988
Agathisanthemum Klotzsch
Amphiasma Bremek.
Anthospermopsis (K. Schum.) J. H. Kirkbr.
Arcytophyllum Willd. ex Schult. &
Schult. f.
Astiella Jovet
Bouvardia Salisb.
Bradea* Standl. ex Brade
Carterella Terrell
Conostomium (Stapf) Cufod.
Crusea Cham. & Schltdl.
Dentella J. R. Forst & G. Forst.
Diacrodon Sprague
Dibrachionostylus Bremek.
Denscantia E. L. Cabral & Bacigalupo
Diodella Small (1)
Diodia L. (1)
Dolichometra K. Schum.
Emmeorhiza Pohl ex Endl.
Ernodea Sw. (2)
Galianthe Griseb. (3)
Gomphocalyx Baker
Hedyotis L.
Hedythyrsus Bremek.
Houstonia L. (4)
Hydrophylax L. f.
Kadua Cham. & Schltdl. (incl. Gouldia
A. Gray and Wiegmannia Meyen) (5)
Kohautia Cham. & Schltdl. (6)
Lelya Bremek.
Leptomischus* Drake
Leptoscela Hook. f.
Lucya DC.
Manettia Mutis ex L.
Manostachya Bremek.
Micrasepalum Urb.
Mitracarpus Zucc. ex Schult.
& Schult. f.
Mitrasacmopsis Jovet
Neanotis W. H. Lewis
Neohymenopogon* Bennet
Nesohedyotis (Hook. f.) Bremek.
Nodocarpaea A. Gray
Oldenlandia L. (incl. Eionitis
Bremek., Exallage Bremek.,
and Thecorchus Bremek.)
Oldenlandiopsis Terrell & W. H. Lewis
Pentanopsis Rendle
Pentodon Hochst.
Phyllocrater Wernham
Phylohydrax Puff
Native distribution
Hed
Hed
Spe
Hed
tropical and S Africa, Comoros
tropical and S Africa
NE Brazil
Mexico to W South America
Hed
Cin/Hed
Hed
Hed
Hed
Spe
Hed
Spe
Hed
Spe
Spe
Spe
Hed
Spe
Spe
Spe
Spe
Hed
Hed
Hed
Spe
Spe
Madagascar
S U.S.A., Mexico to C America
SE Brazil
Mexico
Ethiopia to S Africa
Arizona, New Mexico, Mexico to C America
tropical and subtropical Asia to SW Pacific
Brazil
E Tropical Africa
E Brazil
S U.S.A. to S America
S U.S.A. to S America
Tanzania
S tropical America and Trinidad
Florida, Mexico to C America, Caribbean
S and C America
Madagascar
tropical and subtropical Asia to NW Pacific
C and E tropical Africa
N and C America
India, Sri Lanka, Thailand
Hawaiian Islands to S Pacific
Hed
Hed
Hed
Hed
Hed
Cin/Hed
Hed
Spe
Spe
Africa, Madagascar, and Asia
tropical Africa
Assam to China
NE Brazil
Caribbean
tropical America
C and E tropical Africa
Caribbean
tropical America, naturalized elsewhere
Hed
Hed
Cin/Hed
Hed
Spe
Hed
C and E tropical Africa and Madagascar
tropical and subtropical Asia
E Himalaya, Tibet, SC China, N Indo-China
St. Helena
Cuba
pantropical
Hed
Hed
Hed
tropical and subtropical America
Ethiopia to N Kenya
tropical and S Africa, Arabian Pen., W
Indian Ocean, naturalized in America
Borneo
coastal Tanzania to S Africa, Madagascar
Hed
Spe
No. of
species Sampled
4
7
1
17
yes
yes
no
yes
1
42
5
1
5
14
8
1
1
4
16
5
1
1
4
50
1
ca. 115
2
20
1
28
no
yes
no
no
yes
yes
yes
no
yes
no
yes
no
no
yes
yes
yes
yes
yes
yes
yes
no
yes
31
1
7
1
1
124
3
2
58
yes
yes
no
no
no
yes
yes
no
yes
1
33
3
1
1
ca. 240
yes
no
no
yes
no
yes
1
2
2
no
yes
yes
1
2
no
yes
Volume 96, Number 1
2009
Groeninckx et al.
Phylogeny of Spermacoceae
113
Table 1. Continued.
Genus
Robbrecht,
1988
No. of
species Sampled
Native distribution
Pleiocraterium Bremek.
Polyura* Hook. f.
Pseudonesohedyotis Tennant
Psyllocarpus Mart. & Zucc.
Richardia L.
Hed
Hed
Hed
Spe
Spe
Sacosperma* G. Taylor
Schwendenera K. Schum.
Spermacoce L. (incl. Borreria G. Mey.
and Hemidiodia K. Schum.) (7)
Staelia Cham. & Schltdl.
Stenaria (Raf.) Terrell
Stenotis Terrell
Stephanococcus Bremek.
Synaptantha Hook. f.
Tobagoa Urb.
Tortuella Urb.
Hed
Spe
Spe
tropical Asia
E Himalaya to Assam
Tanzania
Brazil
tropical and subtropical America,
naturalized elsewhere
W and C tropical Africa
Brazil
pantropical
Spe
Hed
Hed
Hed
Hed
Spe
Spe
Mexico and S tropical America
C and E U.S.A. to Mexico, Bahamas
Mexico (Baja California)
WC tropical Africa
Australia
Panama to Tobago
Île de la Tortue (Haiti)
Hed, Hedyotideae; Spe, Spermacoceae s. str.; Cin, Cinchoneae.
5 Bacigalupo & Cabral (1999); (2) 5 Negrón-Ortiz & Hickey (1996);
Terrell et al. (2005); (6) 5 Mantell (1985); (7) 5 Dessein (2003).
* Tentatively included.
(1)
RESULTS
Sequence data from the aligned atpB-rbcL, rps16,
and trnL-F regions were analyzed independently
and in a combined analysis (Table 2). Individual
plastid sequence analyses were topologically congruent. Therefore, only the results from the MP and
BI analysis of the combined matrix are presented
(Figs. 2–4). Compared to the topologies of the individual plastid sequence analyses, the combined
plastid trees show increased resolution and branch
support. The Bayesian tree is somewhat better
resolved than the consensus of the MP analysis,
but more resolved lineages have low posterior
probabilities.
Spermacoceae s.l., as delimited in the introduction
(Table 1), form a well-supported monophyletic group
(jackknife support [JS] 5 100, bootstrap support [BS]
5 100, posterior probability [PP] 5 1), as can be seen
in Figure 2. A highly supported pentamerous-flowered
clade including Dentella J. R. Forst. & G. Forst. and
Pentodon Hochst. (JS 5 100, BS 5 99, PP 5 1) is
resolved as sister to the rest of the tribe (Fig. 2). The
remaining ingroup taxa are part of a clade that lacks
significant jackknife and bootstrap support and has
only weak posterior probability (PP 5 0.84). Within
this clade, stars with Roman numerals I to III are
assigned to the three deeper internal nodes that have
reasonable support. These three clades are discussed
in the following paragraphs.
(3)
5 Cabral (1991);
(4)
4
1
1
9
16
no
no
no
no
yes
2
1
250–300
no
no
yes
14
5
7
1
2
1
1
no
yes
no
no
yes
no
no
5 Terrell (1996);
(5)
5
Clade I in Figure 2 (JS 5 88, BS 5 77, PP 5 1)
includes a Kohautia subg. Kohautia Verdc. clade
sister to a clade that includes Pentanopsis and allied
genera. This Pentanopsis clade (JS 5 95, BS 5 95, PP
5 1) is similar to that proposed by Thulin and Bremer
(2004). However, our larger sampling resulted in a
broader circumscription adding Gomphocalyx, Oldenlandia affinis (Roem. & Schult.) DC., O. herbacea
(L.) Roxb., and O. rosulata K. Schum. Our results
support the monophyly of both Amphiasma Bremek.
(JS 5 98, BS 5 98, PP 5 1) and Phylohydrax (JS 5
93, BS 5 95, PP 5 1).
In clade II (JS 5 88, BS 5 83, PP 5 1) of Figure 2,
all Asian and Micronesian Hedyotis species, except H.
tenelliflora Blume, are part of a strongly supported
Hedyotis s. str. clade (JS 5 100, BS 5 100, PP 5 1),
which is sister to a clade including Agathisanthemum
Klotzsch and its allies. This clade of Asian and
Micronesian Hedyotis species also includes H.
fruticosa L., the type species of the genus. Relationships within this Hedyotis s. str. clade remain mostly
unresolved. Within the Agathisanthemum clade,
Agathisanthemum is paraphyletic to Lelya osteocarpa
Bremek. (JS 5 100, BS 5 99, PP 5 1), both sister to a
lineage of African (Oldenlandia angolensis K. Schum.
and O. goreensis (DC.) Summerh.) and North American (O. uniflora L.) Oldenlandia species (JS 5 100,
BS 5 99, PP 5 1).
In the MP consensus, clade II is sister to clade III
(Figs. 2A, 3A). However, this sister relationship lacks
114
Annals of the
Missouri Botanical Garden
significant jackknife and bootstrap support and is not
recovered in the BI (Figs. 2B, 3B).
Within clade III (Figs. 3, 4), the earlier derived
clades lack significant support values in the MP
consensus (Figs. 3A, 4A) and are collapsed in the BI
(Figs. 3B, 4B). Therefore, relationships between the
different subclades of clade III should be interpreted
with caution. In the following paragraphs, these
subclades are discussed individually.
In Figure 3, the monospecific genus Dibrachionostylus Bremek. is sister to a clade of African
Oldenlandia species (O. echinulosa K. Schum., O.
geophila Bremek., and O. nervosa Hiern). However,
this sister relationship lacks significant jackknife and
bootstrap support (Fig. 3A) and is not supported by
the BI (Fig. 3B). The sister relationship of this clade
with respect to Mitrasacmopsis Jovet and its allies also
lacks support. Mitrasacmopsis, another monospecific
genus in the Hedyotis–Oldenlandia group, is nevertheless highly supported as sister to Hedythyrsus
Bremek. (JS 5 99, BS 5 97, PP 5 1), and both are
sister to O. fastigiata Bremek. (JS 5 99, BS 5 99, PP
5 1).
The genus Kadua Cham. & Schltdl. (including
Oldenlandia biflora L.) is resolved as monophyletic
with moderate jackknife and bootstrap support but
maximum Bayesian posterior probability (JS 5 87, BS
5 86, PP 5 1). The Hawaiian Kadua species are
unresolved with respect to the French Polynesian
species, K. rapensis F. Br. The genus Kadua shares a
most recent common ancestor with all sampled
Australian taxa (O. galioides (F. Muell.) F. Muell., O.
mitrasacmoides F. Muell., and Synaptantha tillaeacea
(F. Muell.) Hook. f.), the Austro-Asian species Hedyotis
tenelliflora, and the African species O. lancifolia
(Schumach.) DC. (JS 5 91, BS 5 86, PP 5 1).
The genus Arcytophyllum is strongly supported as
monophyletic by our analysis (JS 5 93, BS 5 92, PP
5 1). It is sister to a clade of North and Central
American species of Houstonia, Oldenlandia, and
Stenaria (Raf.) Terrell. The Houstonia species plus S.
nigricans (Lam.) Terrell form one clade, although
without significant support.
In Figure 4, Spermacoceae s. str. is nested within
the Hedyotis–Oldenlandia group. In the MP consensus
(Fig. 4A), it forms a monophyletic lineage (although
lacking significant jackknife support and bootstrap
support), while in the BI tree (Fig. 4B), Nesohedyotis
arborea (Roxb.) Bremek. is nested within the
Spermacoceae s. str. clade (although with low PP 5
0.77). In both MP and BI analysis, Spermacoceae s.
str. has uncertain relationships with respect to
Arcytophyllum serpyllaceum (Schltdl.) Terrell, Bouvardia, Manettia, Nesohedyotis (Hook. f.) Bremek.
(Fig. 4A), O. tenuis K. Schum., and O. salzmannii
(DC.) Benth. & Hook. f. ex B. D. Jacks. Sister to this
polytomy is a clade with species of Kohautia subg.
Pachystigma Bremek. and Oldenlandia species,
including the type species O. corymbosa L. (JS 5
99, BS 5 98, PP 5 1). Consequently, species of the
genus Kohautia Cham. & Schltdl. fall in two wellsupported, not closely related clades, which correspond to the two described subgenera: subgenus
Kohautia (JS 5 99, BS 5 99, PP 5 1) and subgenus
Pachystigma (JS 5 96, BS 5 96, PP 5 1).
DISCUSSION
Our analysis corroborates the monophyly of Spermacoceae s.l. (Table 1), a mainly herbaceous assemblage distributed pantropically, with only a few genera
penetrating into more temperate regions. The morphological variation is considerable, but the fimbriate
stipules and tetramerous flowers are shared by most
species. There are no clear morphological synapomorphies that separate Spermacoceae s.l. from its
sister tribe, the emended Knoxieae. The main
differences are listed in Table 3.
Our analyses show several major evolutionary
lineages within Spermacoceae s.l. and allow us to
evaluate the monophyly of a number of genera.
Several genera that have been recognized within the
Hedyotis–Oldenlandia group are supported here as
monophyletic (Amphiasma, Arcytophyllum, Dentella,
Kadua, and Phylohydrax), while others appear to be
paraphyletic (e.g., Agathisanthemum), biphyletic (Kohautia), or polyphyletic (Hedyotis and Oldenlandia
sensu Bremekamp). These groups are discussed in the
following paragraphs.
SPERMACOCEAE S. STR.
In our analyses, Spermacoceae s. str. is nested
within the Hedyotis–Oldenlandia group, which no
longer makes it possible to recognize this lineage at a
tribal level. Additionally, Spermacoceae s. str. as
delimited by Robbrecht (1988) is not corroborated as
monophyletic. Both MP and BI analyses show that it is
necessary to exclude Gomphocalyx and Phylohydrax
for Spermacoceae s. str. to be monophyletic, which is
in agreement with Thulin and Bremer (2004) and
Dessein et al. (2005a).
In the BI analyses, Nesohedyotis arborea, a species
previously included in Hedyotideae, is placed within
Spermacoceae s. str. as sister to Emmeorhiza
umbellata (Spreng.) K. Schum., but lacking significant
posterior probability (PP 5 0.67). This position of
Nesohedyotis within Spermacoceae s. str. was not
recovered in the MP analysis. Because no morphological characters can be found to support Nesohed-
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Figure 1. Floral diversity among species of Spermacoceae. —A. Kohautia microcala Bremek. —B. Hedythyrsus
spermacocinus (K. Schum.) Bremek. —C. Mitracarpus frigidus (Willd. ex Roem. & Schult.) K. Schum. —D. Spermacoce debilis
Benth. —E. Oldenlandia herbacea (L.) Roxb. —F. Gomphocalyx herniarioides Baker. —G. Manostachya ternifolia E. S.
Martins. —H. Oldenlandia lancifolia (Schumach.) DC. —I. Phylohydrax madagascariensis (Willd. ex Roem. & Schult.) Puff.
—J. Manettia luteorubra (Vell.) Benth. —K. Agathisanthemum globosum (Hochst. ex A. Rich.) Klotzsch. —L. Oldenlandia
goreensis (DC.) Summerh. —M. Kohautia coccinea Royle. —N. Oldenlandia biflora L. —O. Kadua acuminata Cham. &
Schltdl. —P. Oldenlandia robinsonii Pit.
yotis as part of Spermacoceae s. str., we suggest that
the difference between the MP and BI analysis could
be the result of data sampling artifacts (only rps16 was
sequenced for N. arborea), which probably affected
the BI more than the MP analysis.
With the deeper nodes unresolved or only weakly
supported, the relationships within Spermacoceae s.
str. remain unclear and should be the subject of
further phylogenetic studies including more taxa and/
or characters. Nevertheless, our analyses corroborate
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Table 2. Characteristics of each data matrix and the corresponding tree statistics.
atpB-rbcL
rps16
trnL-trnF
Combined
No. of taxa
No. of char.
No. of PI char.
No. of PI indels
No. of MPT
MPT length
CI
RI
100
105
107
128
1237
705
1053
2995
175
191
184
550
31
20
29
80
1949
1351
343
4782
399
525
423
1385
0.55
0.56
0.62
0.56
0.84
0.82
0.88
0.84
Char, characters; CI, consistency index (Kluge & Farris, 1969); MPT, most parsimonious tree(s); PI, potentially informative;
RI, retention index (Farris, 1989).
the monophyly of most of the commonly accepted
genera within Spermacoceae s. str., notably Crusea
Cham. & Schltdl., Mitracarpus Zucc. ex Schult. &
Schult. f., and Richardia L., although these were
sampled only with a few species. In contrast, the two
Galianthe Griseb. sampled species are paraphyletic
to Diodia spicata Miq., a species that was recently
excluded from Diodia s. str. and transferred to
Borreria. If the position of D. spicata is confirmed by
further phylogenetic studies, the generic circumscription of Galianthe should be widened to include
at least this species. Dessein (2003) already showed
that palynological data (7-zonocolporate pollen, long
ectocolpi, double reticulum) support a close relation
between D. spicata and Galianthe. Diodia L. as
traditionally delimited, including species referred to
Diodella Small by Bacigalupo and Cabral (1999), is
not supported as monophyletic. Also, Spermacoce
s.l., including Borreria, is not supported as monophyletic.
BOUVARDIA AND MANETTIA
Manettia is strongly supported as monophyletic (JS
5 100, BS 5 100, PP 5 1), whereas support for
Bouvardia is moderate (JS 5 85, BS 5 87, PP 5
0.99). In accordance with Andersson et al. (2002),
Arcytophyllum serpyllaceum is corroborated as sister
to Bouvardia. This strongly supported relationship (JS
5 99, BS 5 99, PP 5 1), in combination with the fact
that the remaining Arcytophyllum species are strongly
supported as a monophyletic and distinct lineage (see
below), suggests that at least A. serpyllaceum should
be included within Bouvardia. Although Bouvardia is
generally considered as a genus of shrubs only, it
comprises both subshrubs and perennial herbs
(Blackwell, 1968), which makes it possible to fit in
A. serpyllaceum. Arcytophyllum serpyllaceum is similar
to Bouvardia and different from other Arcytophyllum
species in many respects. First, the stipule margin of
A. serpyllaceum is not dentate or fimbriate, as in most
Arcytophyllum species (Mena, 1990), but consists of a
basal sheath and a trullate mucro as in most
Bouvardia species (Blackwell, 1968). Second, whereas the seeds of Arcytophyllum are more or less
cymbiform (Mena, 1990), those of A. serpyllaceum are
discoid with a central hilum as in Bouvardia
(Andersson et al., 2002). The major difference
between seeds of A. serpyllaceum and Bouvardia is
that Bouvardia seeds are winged, whereas those of A.
serpyllaceum are not.
ARCYTOPHYLLUM–HOUSTONIA CLADE
Previous studies based on plastid DNA sequences
have shown Arcytophyllum to be monophyletic and
closely related to the North American Houstonia
(Andersson & Rova, 1999; Andersson et al., 2002).
Our analyses support the monophyly of the Neotropical genus Arcytophyllum (JS 5 93, BS 5 92, PP 5 1)
only if A. serpyllaceum is excluded from the genus (see
above). Sister to Arcytophyllum is a group of North and
Central American species presently classified in the
genera Houstonia, Oldenlandia, and Stenaria. By
having its closest relatives in North America rather
than in South America, Arcytophyllum may be one of
the few examples within Rubiaceae that has reached
the Andes by a southern migration (Andersson et al.,
2002). From this perspective, Mesoamerican species
like O. microtheca (Cham. & Schltdl.) DC. may
represent remnants of stepping-stone populations.
The Arcytophyllum–Houstonia clade as defined by
our results is thus restricted to the New World. Seeds
of Arcytophyllum and Houstonia are generally more or
less cymbiform. Our results thus support Schumann’s
(1891) grouping of genera with cymbiform seeds. So
far, Neanotis W. H. Lewis has not been sequenced,
but if seed shape is indeed a good phylogenetic
marker, Neanotis could be the closest non-American
relative of the Arcytophyllum–Houstonia clade (Lewis,
1966).
There has been much discussion about the
recognition of Houstonia at the generic level. In a
recent molecular study based on ITS and trnL intron
data (Church, 2003), Houstonia appeared to be
paraphyletic with respect to the North American
genus Stenaria. Therefore, Church (2003) suggested
that Houstonia and Stenaria are better treated as a
single genus. As currently circumscribed (Terrell,
1996), the genus Houstonia is composed of 20 species
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Figure 2. —A. Part one of the strict consensus tree of the 4782 MPTs from the combined analysis including atpB-rbcL, rps16, and trnL-trnF sequences (L 5 1385, consistency index [CI] 5
0.56, retention index [RI] 5 0.84). Jackknife (left) and bootstrap (right) values (. 50) are indicated above branches. —B. Part one of the Bayesian tree based on combined atpB-rbcL, rps16, and
trnL-trnF data. Posterior probabilities are indicated above branches. The tribe Spermacoceae s.l. starts on Figure 2 (as indicated by an arrow) and continues over to Figures 3 and 4. Stars with
Roman numerals I to III are assigned to the three reasonably supported clades within Spermacoceae s.l.
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Figure 3. —A. Part two of the strict consensus tree of the 4782 MPTs from the combined analysis including atpB-rbcL, rps16, and trnL-trnF sequences (L 5 1385, CI 5 0.56, RI 5 0.84).
Jackknife (left) and bootstrap (right) values (. 50) are indicated above branches. —B. Part two of the Bayesian tree based on combined atpB-rbcL, rps16, and trnL-trnF data. Posterior
probabilities are indicated above branches. Roman numeral III is assigned to the deeper internal node with reasonable support. The tribe Spermacoceae s.l. starts on Figure 2 and continues over
to Figures 3 and 4. Stars with Roman numerals I to III are assigned to the three reasonably supported clades within Spermacoceae s.l.
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119
Figure 4. —A. Part three of the strict consensus tree of the 4782 MPTs from the combined analysis including atpB-rbcL, rps16, and trnL-trnF sequences (L 5 1385, CI 5 0.56, RI 5 0.84).
Jackknife (left) and bootstrap (right) values (. 50) are indicated above branches. —B. Part three of the Bayesian tree based on combined atpB-rbcL, rps16, and trnL-trnF data. Posterior probabilities are
indicated above branches. The tribe Spermacoceae s.l. starts on Figure 2 and continues over to Figures 3 and 4. Representatives of the former tribe Spermacoceae s. str. are shown in this portion of the
strict consensus tree and the Bayesian tree.
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Table 3. Major morphological differences between Knoxieae and Spermacoceae s.l.
Knoxieae s.l.
Spermacoceae s.l.
Merosity
Inflorescence
Calyx lobes
Pollen
often 5-merous or derived from the 5-merous state
terminal (including pseudoaxillary)
often 1 or more calyx lobes enlarged
bireticulum not yet reported
Exotesta
Distribution
ITW often slightly thickened
paleotropical, centered in Madagascar and
continental Africa
often 4-merous, rarely 5-merous
terminal or axillary
rarely enlarged calyx lobes
bireticulum common, often associated with
heterostyly
ITW without thickenings
pantropical, with a few taxa reaching outside
the tropics
ITW, inner tangential wall.
restricted to North America. The genus contains both
annual and perennial herbs with either heterostylous
or homostylous flowers, crateriform seeds, and
colporate pollen. Chromosome numbers are variable
among species of the genus with x 5 6, 7, 8, or 11.
Stenaria, a genus only recently described (Terrell,
2001a), includes five species previously included in
the North American Hedyotis. The genus contains only
perennial, heterostylous herbs. Due to our incomplete
sampling of these two genera, and given that
Houstonia forms a polytomy with Stenaria, our results
are not conclusive with respect to whether it is best to
recognize Stenaria or consider it part of a more
broadly delimited Houstonia. A more extensive
sampling should focus further on this question.
Sister to the Houstonia–Stenaria clade is Oldenlandia microtheca. The prevailing basic chromosome number in Oldenlandia is n 5 9, which occurs
in the type species O. corymbosa and in many of the
species native to North America, Asia, Africa, and
Australia (Lewis, 1965), but not in O. microtheca,
which is exceptional in having a chromosome number
n 5 11. The same chromosome number is found in
Oldenlandiopsis Terrell & W. H. Lewis (Terrell,
1991), not included in this study, and in some
Houstonia species (e.g., H. rubra Cav.). Until now,
Oldenlandia microtheca and Oldenlandiopsis were
never considered to be closely related to Houstonia
because of the lack of morphological similarities
(Lewis, 1965; Terrell, 1991).
Oldenlandiopsis contains only one species, O.
callitrichoides (Griseb.) Terrell & W. H. Lewis,
previously included in Oldenlandia. This smallleaved, small-flowered, creeping herb is native to
the West Indies and southern Mexico. Based on its
chromosome number and its distribution, a position of
Oldenlandiopsis in the Arcytophyllum–Houstonia
clade close to Oldenlandia microtheca seems quite
likely. However, seeds of Oldenlandiopsis are noncrateriform and pollen are 8-colporate with a
lalongate, slightly crassimarginate endoaperture (Terrell & Lewis, 1990). These types of seeds and pollen
are unusual within the Arcytophyllum–Houstonia
clade. Plurizonocolporate pollen grains are also
exceptional within the rest of the Hedyotis–Oldenlandia group, where the aperture number rarely
exceeds five. The Asian genus Neanotis (Lewis, 1966),
the Malagasy endemic Gomphocalyx (Dessein et al.,
2005a), the Afro-Madagascan Phylohydrax (Puff,
1986), and the West Indian monotypic genus Lucya
DC. (Terrell & Lewis, 1990) are notable exceptions
within the Hedyotis–Oldenlandia group in having
plurizonocolporate pollen grains. Both Gomphocalyx
and Phylohydrax belong to the Pentanopsis clade
(see below). With no molecular sequence data
available for Lucya, Neanotis, and Oldenlandiopsis,
it would be premature to hypothesize a close
relationship between any of these taxa and the
Arcytophyllum–Houstonia clade or the Pentanopsis
clade. Nevertheless, considering their distribution, the
Caribbean-Mexican genera Lucya and Oldenlandiopis
are more likely to fall in the Arcytophyllum–Houstonia
clade, whereas the Asian genus Neanotis is more
likely to have its closest relatives within the
Pentanopsis clade.
Two closely related genera from Baja California,
Stenotis Terrell (Terrell, 2001b) and Carterella Terrell
(Terrell, 1987), may also belong to the Arcytophyllum–
Houstonia clade. Like the Mesoamerican species
Oldenlandia microtheca, they may represent remnants
of stepping-stone populations. The monospecific
genus Carterella was described based on Bouvardia
alexanderae A. M. Carter. It resembles Bouvardia in
having unusually long corolla tubes, but differs from
Bouvardia in having wingless seeds and chromosome
number n 5 13. The genus Stenotis, on the other
hand, includes seven former Hedyotis species endemic
to the Baja California peninsula (Terrell, 2001b).
These heterostylous, annual or perennial herbs also
have chromosome number x 5 13. According to
Terrell (1987, 2001b), Carterella and Stenotis have
their closest relatives among the Baja California
species of Houstonia (H. mucronata group sensu
Terrell et al., 1986).
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KADUA
which includes the type species H. fruticosa (Sri
Lanka). Several authors already considered the genus
Hedyotis to be a distinct Asian taxon (Bremekamp,
1952; Hallé, 1966; Terrell, 1975, 1991; Andersson et
al., 1999). Hedyotis fruticosa and its Asian relatives
are not closely related to the American species of
Hedyotis (Houstonia lineage) or to the Polynesian
species (Kadua). The Asian and Micronesian Hedyotis
species (Hedyotis s. str.) differ from the American and
Polynesian ones in their combination of a robust
(sometimes shrubby) habit, small beaked and diplophragmous capsules, dorsiventrally compressed seeds
with the hilum on a conspicuous central ridge (Terrell
& Robinson, 2003), and a high chromosome number
(Kiehn, 1986). Our results clearly demonstrate that a
broad concept of Hedyotis, merging several genera
(Hedyotis s. str., Houstonia, Kadua, Kohautia, Oldenlandia, etc.), as was proposed by several researchers (Fosberg, 1943; Merrill & Metcalf, 1946; Rogers,
1987; Wagner et al., 1989; Fosberg & Sachet, 1991;
Dutta & Deb, 2004), is no longer supported. If this is
confirmed with further sampling, all North American
species now called Hedyotis would require new
combinations under other generic names.
Pleiocraterium Bremek. (not included in this study)
is probably related to the Hedyotis s. str. clade. The
genus was described by Bremekamp in 1939, including
four species distributed in India, Sri Lanka, and
Sumatra. The generic name refers to the numerous
cups that are formed by the connate leaf bases. The
type species of the genus, P. verticillare (Wall. ex Wight
& Arn.) Bremek., was previously included in Hedyotis.
However, the genus differs from other Hedyotis s. str.
species in having distinctly beaked capsules and
parallel-nerved, quaternate leaves. The internodes
remain very short, as a result of which the leaf whorls
are clustered in rosettes. It will be necessary to wait,
however, until molecular data of Pleiocraterium
become available before a close relation of the genus
to the Asian Hedyotis species is confirmed.
Our results support the resurrection of the genus
Kadua for the Polynesian Hedyotideae (Hawaiian
Islands and French Polynesia: Terrell et al., 2005).
This taxonomic change was previously suggested by
unpublished molecular data (Motley et al., 1998;
Motley, 2003) and by morphological studies of the
seed anatomy of the Hawaiian species (Terrell et al.,
2005). The genus Kadua was treated as a distinct
genus until Fosberg’s (1943) revision of the group. He
included the genus within a broadly delimited
Hedyotis, except for the fleshy-fruited species, which
he treated as Gouldia A. Gray (Fosberg, 1937). Kadua
species can, however, easily be distinguished from
other Hedyotis species by their salverform, fleshy
corollas with appendaged lobes, and by their either
tardy, often incomplete septicidal dehiscent capsules
or indehiscent drupaceous fruits (Terrell et al., 2005).
The genus Kadua currently comprises 28 species; all
are indigenous to the Pacific Islands with the majority
(21 species) occurring on the Hawaiian Islands
(Terrell et al., 2005). Seeds of these Hawaiian Kadua
species fall into four groups, described by Terrell et
al. (2005). Based on the chloroplast data alone, the
relationships within the genus Kadua remain mostly
unresolved. Only section Wiegmannia Meyen, W. L.
Wagner & Lorence (represented in our sampling by K.
cordata Cham. & Schltdl., K. degeneri (Fosberg) W. L.
Wagner & Lorence, K. elatior (H. Mann) W. L.
Wagner & Lorence, K. flynnii (W. L. Wagner &
Lorence) W. L. Wagner & Lorence, K. laxiflora H.
Mann, K. littoralis Hillebr., and K. parvula A. Gray)
and section Gouldiopsis (Fosberg) W. L. Wagner &
Lorence (represented in our sampling by Kadua
centranthoides Hook. & Arn. and K. foggiana
(Fosberg) W. L. Wagner & Lorence) were recovered.
A broader sampling including more Kadua species
and more molecular markers is needed to discuss
molecular evolution in the light of the seed morphological observations of Terrell et al. (2005).
Oldenlandia biflora is sister to the Kadua clade. Its
distribution from (sub)tropical Asia to the western
Pacific is consistent with the sister relationship to the
Polynesian Kadua clade. Our results show that O.
biflora can no longer be included within the genus
Oldenlandia, but it is necessary to await further
studies before transferring it to Kadua or describing a
new genus. So far, we have not found morphological
characters to support the transfer.
HEDYOTIS S. STR.
It seems appropriate to restrict the name Hedyotis to
the Asian and Micronesian species of the genus,
121
AGATHISANTHEMUM CLADE (CLADE II)
The African genus Agathisanthemum is not supported as monophyletic by our analyses. The monotypic African genus Lelya Bremek. is nested within
Agathisanthemum, making it paraphyletic as currently
circumscribed and suggesting that Lelya should be
reduced to Agathisanthemum. This proposal is
supported by several palynological characters. Scheltens (1998) showed that Agathisanthemum and Lelya
share the same pollen type, characterized by a distinct
endocolpus or endocingulum, a mesoporus surrounded
by a costa at the inside of the grain (described as a
compound ora by Lewis, 1965), and a microreticulate
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sexine with granules on the muri facing the lumina
(bireticulum).
A group of African Oldenlandia species is sister to
Agathisanthemum. Two of the three Oldenlandia
species, O. angolensis and O. goreensis, belong to
Oldenlandia subg. Anotidopsis (Hook. f.) K. Schum.
This subgenus, as described by Bremekamp (1952),
includes three other putative species of which only O.
cephalotes (Hochst.) Kuntze (not included in our
sampling) is currently recognized. Subgenus Anotidopsis is distributed in Asia, Australia, and Africa and is
characterized by distinctly beaked capsules. The New
World taxon O. uniflora is sister to O. angolensis and O.
goreensis. More detailed (molecular as well as morphological) studies within the Agathisanthemum clade are
needed to evaluate if the three Oldenlandia species, O.
angolensis, O. goreensis, and O. uniflora, or the entire
Oldenlandia subg. Anotidopsis, are to be transferred to
a new genus or if these species are better treated as
members of the genus Agathisanthemum.
The Asian Hedyotis species are sister to the
Agathisanthemum–Oldenlandia clade. This relationship is not unexpected as Bremekamp (1952) already
suggested a close relationship between Agathisanthemum and the Asian Hedyotis species (i.e., Hedyotis
sect. Diplophragma) based on a similar type of
dehiscence of the capsules.
Thulin & Bremer, 2004; Dessein et al., 2005a). The
close relationship between Gomphocalyx and Phylohydrax is supported by our results and by several
morphological characters (amphistomatic leaves, plurizonocolporate pollen, indehiscent fruits, and seeds
with a weak, pale exotesta) as shown by Dessein et al.
(2005a). Almost all taxa in the Hedyotis–Oldenlandia
group have multiovulate ovaries, and the number of
pollen apertures rarely exceeds five. The presence of
uniovulate ovaries and plurizonocolporate pollen were
the main reasons why Gomphocalyx and Phylohydrax
were previously included in Spermacoceae s. str.,
where it is more common than in the rest of the
Spermacoceae s.l. tribe, in which 3-colporate pollen
predominates (Dessein et al., 2002, 2005b; Dessein,
2003). As mentioned above, the Asian genus Neanotis
is a notable exception in having plurizonocolporate
pollen grains. The genus also shows a trend toward
reduction in the number of seeds per locule. In mature
fruits, only one or two seeds are present. However,
with no molecular sequence data available for the
genus it would be premature to hypothesize a close
relationship between Neanotis, Gomphocalyx, and
Phylohydrax. A few authors (Capuron, 1973; Piesschaert, 2001) also proposed a close relationship
between Gomphocalyx and Lathraeocarpa Bremek.,
another endemic to Madagascar. Although Lathraeocarpa is not a trailing herb like Gomphocalyx but a
(sub)shrub, the two taxa share a calyx with eight lobes,
uniovulate ovaries, and plurizonocolporate pollen. The
last two characters also support a close relationship
between Phylohydrax and Lathraeocarpa. However,
several morphological characters distinguish Lathraeocarpa from Gomphocalyx, some of which might
even point to an affinity with Triainolepis Hook. f.
First, the (sub)shrubby habit of Lathraeocarpa is much
more similar to the shrubby habit of Triainolepis than
to the herbaceous habit of Gomphocalyx. Second, the
pyrene of L. decaryi Bremek. is surrounded by eight
strands of thin-walled cells, a condition very similar to
that observed in some Triainolepis species (Bremekamp, 1957; Piesschaert, 2001). Likewise, Lathraeocarpa and Triainolepis have a plurilocular ovary and
fleshy fruits, whereas Gomphocalyx has a bilocular
ovary and dry fruits, which has prompted some authors
(Kårehed & Bremer, 2007) to tentatively include
Lathraeocarpa in the emended tribe Knoxieae rather
than in Spermacoceae s.l. However, we will have to
wait until molecular data become available to assess
the taxonomic position of Lathraeocarpa with more
certainty (Dessein et al., 2005a).
Species of Conostomium form a strongly supported
clade (JS 5 99, BS 5 99, PP 5 1) together with
Oldenlandia herbacea. The type of the genus Conostomium, C. natalense (Hochst.) Bremek., is unre-
PENTANOPSIS CLADE
Our sampling resulted in a broader concept of the
Pentanopsis clade than proposed by Thulin and
Bremer (2004). They included Amphiasma, Conostomium (Stapf) Cufod., Manostachya Bremek., Pentanopsis, and Phylohydrax.
Oldenlandia affinis was not included in the study of
Thulin and Bremer (2004), but it was shown to be
closely related to the African genus Amphiasma by
Andersson and Rova (1999) and Dessein et al.
(2005a). Amphiasma, O. affinis, and Pentanopsis share
sessile linear leaves, indistinctly beaked capsules,
non-mucilaginous seeds and nonpunctate testa cells
(Bremekamp, 1952). However, a detailed study is
needed to find more unambiguous morphological
characters to support a relation among the three taxa.
In the past, Gomphocalyx (a monospecific genus
endemic to Madagascar) and Phylohydrax (a genus
described in 1986 by Puff to accommodate the East
African and Madagascan Hydrophylax L. f. species)
were both included in Spermacoceae s. str. based on
their uniovulate ovaries and plurizonocolporate pollen
grains (Robbrecht, 1988). However, recent molecular
studies excluded both genera from Spermacoceae s.
str. and suggested that they are closely related to one
another and to the Pentanopsis clade (Dessein, 2003;
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Phylogeny of Spermacoceae
solved with respect to the other species of Conostomium and to O. herbacea. Both Conostomium and O.
herbacea have seeds with coarsely granulate testa
cells (Bremekamp, 1952; Dessein, 2003) and pollen
that is larger than that of most other genera within the
Hedyotis–Oldenlandia group (Bremekamp, 1952;
Scheltens, 1998). These characters, however, are
homoplasious because granulate testa cells and large
pollen grains also occur elsewhere in the Hedyotis–
Oldenlandia group. We observed granulate testa cells
in Kohautia subg. Pachystigma, O. corymbosa, and O.
nematocaulis Bremek., whereas large pollen grains are
characteristic of Amphiasma, Gomphocalyx, and
Phylohydrax. The most striking feature of Conostomium pollen, namely the short ectocolpi (Scheltens,
1998; Dessein et al., 2005a), is not found in O.
herbacea or in most other members of the Pentanopsis
clade, but it is reported for Gomphocalyx and
Phylohydrax (Dessein et al., 2005a).
The last additional species falling in the Pentanopsis clade is Oldenlandia rosulata, an African
species named after its basal rosulate leaves. The
relationship of O. rosulata to other members of the
Pentanopsis clade remains unclear.
Despite the strong support for the Pentanopsis clade
(JS 5 95, BS 5 95, PP 5 1) in our molecular analyses,
the group is not easily morphologically characterized.
The only unifying feature for the clade would be what
Thulin and Bremer (2004) called basal placentation.
Nevertheless, the placentation is not truly basal, but
rather axile with the placenta or ovule attached near the
base of the septum. Our observations show that this
kind of placentation is also found outside the
Pentanopsis clade. Moreover, the basal placentation
character state is only vaguely defined, and more
detailed placentation studies within Spermacoceae s.l.
are needed before further conclusions can be drawn
about the phylogenetic value of this character.
relationship of this monospecific genus to Hedythyrsus
and Oldenlandia fastigiata. Our own observations
have identified similar placentation types within these
taxa. Moreover, Hedythyrsus and Mitrasacmopsis have
the same type of capsule dehiscence (loculicidal
followed by septicidal dehiscence), seeds with testa
cells that show the same undulating radial walls, and
pollen with a double reticulum (Groeninckx, 2005).
The monospecific genus Dibrachionostylus is sister to
a clade of African Oldenlandia species. The genus was
separated from Oldenlandia largely on the basis of its
capsule dehiscence (both loculicidal and septicidal vs.
only loculicidal in Oldenlandia). Bremekamp (1952)
closely associated Dibrachionostylus with Agathisanthemum because of their similar fruit dehiscence. However,
Dibrachionostylus differs markedly from Agathisanthemum in the pollen aperture morphology (Lewis, 1965).
As mentioned above, Agathisanthemum has a distinct
ectocolpus, an endocolpus or endocingulum, and a
mesoporus surrounded by a costa at the inside of the
grain (Lewis, 1965). Pollen grains of Dibrachionostylus
are also 3-colporate but do not have a costa on the inside
(Lewis, 1965). The apertures of Dibrachionostylus
pollen are, therefore, more similar to those of
Amphiasma, Oldenlandia, and Pentodon (Lewis, 1965).
Nesohedyotis is another monospecific genus previously included in the Hedyotideae. Its only species, N.
arborea, shows a superficial resemblance to the East
African genus Hedythyrsus; specimens of both taxa turn
black when dried, and their leaf shape and inflorescence
structure are similar (Bremekamp, 1952). However, our
results show that Nesohedyotis is more closely related to
the former tribes Spermacoceae s. str. and Manettieae
than to members of the Hedyotis–Oldenlandia group.
Nesohedyotis has unisexual flowers, which are unusual
among Spermacoceae, and, in contrast to Hedythyrsus,
its fruits open by a single loculicidal split. Although it is
one of the more common endemic species on St. Helena,
its small population size and small geographical
distribution make Nesohedyotis Endangered (EN) according to IUCN Red List criteria (IUCN, 2001).
According to Verdcourt (1976), the monospecific
Tanzanian Pseudonesohedyotis Tennant, which is not
included in our sampling, is closely related to
Nesohedyotis and Hedythyrsus. Pseudonesohedyotis
has indeed the same leaf shape and inflorescence
structure as the latter two taxa. In habit and
distribution, however, it resembles Hedythyrsus more
than Nesohedyotis. Both Pseudonesohedyotis and
Hedythyrsus are (sub)shrubs, whereas Nesohedyotis is
a small tree. Moreover, Pseudonesohedyotis differs
from Nesohedyotis in having hermaphroditic flowers.
Again, it is necessary to wait until molecular data
become available to discuss the taxonomic position of
Pseudonesohedyotis with more confidence.
MONOSPECIFIC GENERA WITHIN THE HEDYOTIS–
OLDENLANDIA GROUP
Besides the genus Gomphocalyx of the Pentanopsis
clade, the Hedyotis–Oldenlandia group comprises
several other monospecific genera. These monospecific genera often have several peculiar characters,
making it very difficult to discuss their relationship
with other Spermacoceae.
In our sampling, for example, the Afro-Madagascan
genus Mitrasacmopsis has seeds with undulating
radial exotesta cell walls, distinctly stalked placentas
with ovules positioned on the periphery of the
placental tissue, pollen grains with a double reticulum, and fruits with a conspicuous beak (Groeninckx
et al., 2007). Our molecular results suggest a close
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Based on the presence of an apparently superior
ovary, Jovet (1941) originally placed Mitrasacmopsis
and Astiella Jovet, another monospecific genus of the
Hedyotis–Oldenlandia group endemic to Madagascar
(not included in this study), within Loganiaceae–
Spigelieae. Members of Rubiaceae are generally
characterized by the presence of an inferior ovary.
Groeninckx et al. (2007) demonstrated that flowers of
Mitrasacmopsis are initially epigynous with inferior
ovaries. Expansion of the upper part of the ovary in
fruiting stage results in a change in the ovary position
of Mitrasacmopsis from basically inferior to secondarily semi-inferior. The same kind of fruit development
also most likely occurs in Astiella. In her morphological study of the Rubioideae, Hayden (1968) stated
that some genera of Spermacoceae s. str. have semiinferior fruits. According to Robbrecht (1988), this
statement is based on the strong expansion of the top
of the nectary disc in the fruiting stage. However, we
have not observed semi-inferior ovaries within
Spermacoceae s. str. Nevertheless, within Spermacoceae s.l. several other taxa, apart from Mitrasacmopsis
and Astiella, are characterized by the presence of a
beak at fruit stage (Conostomium spp., Hedythyrsus
spp., Kohautia spp., Oldenlandia spp.). These beaks
are not remnants of the nectary disc and probably
originate in a similar way as in Mitrasacmopsis.
However, the ovaries of these species do not undergo a
remarkable reverse in shape in the fruiting stage as
observed in Mitrasacmopsis and Astiella. Based on
their fruit shape, Mitrasacmopsis and Astiella seem
closely related. However, Jovet (1941) also suggested
a close relationship between Astiella and the Asian
Anotis DC. species, presently classified in the genus
Neanotis (Lewis, 1966). Astiella differs from both
genera in having only two calyx lobes, a character that
so far has not been observed within the Hedyotis–
Oldenlandia group, and uniovulate locules. Molecular
sequence data of Astiella will allow us to discover the
taxonomic position of the genus in the future.
Other monospecific genera of the Hedyotis–Oldenlandia group are Carterella, Dolichometra K.
Schum., Lelya, Leptoscela Hook. f., Lucya, Phyllocrater Wernham, Polyura Hook. f., Stephanococcus
Bremek., and Oldenlandiopsis. The genera Carterella,
Lelya, Lucya, and Oldenlandiopsis were already
discussed in previous sections. To date, the taxonomic
position of most of these monospecific genera remains
controversial because molecular data are lacking.
Egypt, and throughout most of Africa south of the
Sahara (including Socotra, Cape Verde, and Madagascar). The genus can easily be distinguished from
other representatives of the Hedyotis–Oldenlandia
group by its unique flower morphology. The anthers
and stigma are always included, with the stigma held
well below the anthers or occasionally just touching
them. This monomorphic short-styled condition is,
with the exception of a few individuals of Conostomium, unique among the African members of the
former tribe Hedyotideae. For this reason, Kohautia
has always been considered a distinct genus (Bremekamp, 1952; Mantell, 1985). Our molecular results,
however, show that the two subgenera of Kohautia are
not sister clades. Subgenus Kohautia is sister to the
Pentanopsis clade, whereas subgenus Pachystigma is
sister to an Oldenlandia clade containing the type
species O. corymbosa.
Despite the unifying floral architecture, there are
numerous morphological differences between the two
subgenera (Lewis, 1965; Mantell, 1985). The number
of stigmatic lobes is the most striking diagnostic
character that allows identification of the subgenera
even in the field. Members of subgenus Kohautia have
styles with two thin filiform stigma lobes, whereas
Pachystigma is characterized by the presence of only
a single, ovoid to cylindrical stigma lobe. Seeds are
also different in the two subgenera: subgenus
Kohautia seeds are angular-conic to subconic in
shape with 5- or 6-angled testa cells, whereas in
subgenus Pachystigma the seeds are rounded with
wavy and punctate testa cells. Pollen of Kohautia can
also be divided into two easily recognizable groups
coinciding with the two subgenera (Lewis, 1965).
Other differences between the two subgenera are
found in floral architecture and chromosome number.
Based on these differences, Mantell (1985) hypothesized that the two subgenera may have diverged and
developed independently of one another fairly early
on and she even tentatively proposed the elevation of
the two subgenera to generic rank. At that time,
Mantell decided to maintain a widely defined genus
Kohautia, mainly for practical reasons. However, our
molecular data now clearly support the recognition of
two genera. Sampling within the genus still needs to
be improved before proposing new generic circumscriptions.
KOHAUTIA
Kohautia is a genus of 31 species (Mantell, 1985)
distributed from the Indian subcontinent through
Pakistan, Iran, the Arabian Peninsula, Sinai, eastern
OLDENLANDIA
Govaerts et al. (2006) currently accept 76 Oldenlandia species from Africa, 155 from Asia and
Australia, 23 from America, and eight from the Pacific
Islands. However, as documented in previous molecular studies (Bremer, 1996; Andersson & Rova, 1999;
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Phylogeny of Spermacoceae
125
Bremer & Manen, 2000), Oldenlandia is shown to be
polyphyletic.
Bremekamp (1952) divided the 61 species that he
recognized from Africa into 16 subgenera. Our results
do not support the majority of these subgenera. Only
the subgenus Hymenophyllum Bremek. (Oldenlandia
echinulosa and O. nervosa) and subgenus Anotidopsis
(O. angolensis and O. goreensis) are corroborated.
The type species, Oldenlandia corymbosa, is sister
to a clade with the African species O. capensis L. f., O.
robinsonii Pit., O. nematocaulis, O. taborensis Bremek., and O. wauensis Schweinf. ex Hiern. The last
species, O. wauensis, was segregated by Bremekamp
(1952) in a new genus Thecorchus Bremek., which he
proposed to be allied with Otomeria of the tribe
Knoxieae because of its distinctly elongated capsules
and equal number of tetramerous and pentamerous
flowers. However, Kårehed and Bremer (2007) showed
that Thecorchus is not related to Otomeria but is close
to Oldenlandia. Our results, which place Thecorchus
in a clade comprised of the type species of Oldenlandia, support the transfer of T. wauensis
(Schweinf. ex Hiern) Bremek. back into Oldenlandia.
The type species O. corymbosa and O. capensis belong
to Bremekamp’s (1952) subgenus Oldenlandia K.
Schum. Besides these two species, subgenus Oldenlandia also includes O. fastigiata and O. herbacea.
These species are apparently not related to O.
corymbosa and its allies. Oldenlandia fastigiata is
sister to Hedythyrsus and Mitrasacmopsis, whereas O.
herbacea in the Pentanopsis clade is sister to a
paraphyletic Conostomium. Bremekamp (1952) already pointed out that O. herbacea differs from the rest
of the subgenus by the coarsely granulated walls of the
testa cells, the rather large flowers, and the slender
corolla tube.
The Australian species of Oldenlandia, O. mitrasacmoides and O. galioides, sampled here belong to a
clade comprising the Australian Synaptantha tillaeacea, the Austro-Asian O. tenelliflora, the African
species O. lancifolia, and the Kadua species (including O. biflora). Oldenlandia mitrasacmoides is sister to
the rest of the clade. Synaptantha tillaeacea is sister
to a clade with Oldenlandia tenelliflora, O. galioides,
and O. lancifolia. Synaptantha Hook. f. may be
distinguished from the other genera in the clade by its
slightly connate corolla lobes, stamens with filaments
attached to both the corolla and the ovary, depressed
obconic or ovoid seeds, and half-inferior ovaries
(Halford, 1992). In his review of Australian Oldenlandia, Halford (1992) distinguished five groups
mostly based on seed morphology. Oldenlandia
galioides and O. tenelliflora are placed together in
his group one, which is characterized by obconic
seeds that are slightly laterally compressed and
obtriangular in outline. Oldenlandia mitrasacmoides
belongs to his group two, which is characterized by
scutelliform seeds that are oblong or broadly elliptic
in outline, with the hilum situated on a conspicuous
central ridge. The African species O. lancifolia has
seeds similar in shape to those of its sister O. galioides
(Dessein, 1998).
Not all American Oldenlandia species included in
our sampling are placed within the Arcytophyllum–
Houstonia clade (see discussion above). The remaining South American species of Oldenlandia, O.
salzmannii and O. tenuis, form a clade sister to the
former tribes Spermaococeae s. str. and Manettieae.
Terrell (1990) already reported that O. salzmannii is
clearly distinct from Houstonia and Oldenlandia. In
contrast to other Oldenlandia species, O. salzmannii
does not have the typical oldenlandioid seeds or base
number of chromosomes (n 5 15 instead of 9).
Moreover, it shares some unusual characters with
Oldenlandiopsis: stipules are minute, not more than
0.5 mm long (Oldenlandia stipules are often 2–3 mm
long); few stiff hairs occur on the leaves (Oldenlandia
species usually have smaller, softer hairs); and it has a
creeping habit (which is rare in Oldenlandia, the
usual habit being erect to spreading or prostrate). It
would be very informative to include Oldenlandiopsis
in future studies to investigate its relationship to
either O. microtheca (see discussion of the Arcytophyllum–Houstonia clade above) or O. salzmannii.
FUTURE RESEARCH PLANS AND CONCLUSIONS
Although our analyses found well-supported clades
within Spermacoceae s.l., many relationships within
and between these clades still remain unresolved.
Furthermore, many relationships detected here are
contradictory to previous taxonomic treatments and
await morphological backup. This study was a multipartner collaboration resulting in a framework for
future Spermacoceae research. Further studies will
focus on obtaining additional DNA markers (i.e.,
nuclear DNA data) to provide better resolution within
the tribe. Besides improving the character sampling,
we also need to balance the taxon sampling by
including more Asian and American taxa. In addition,
concerted studies will focus on the morphological
characterization of monophyletic groups within Spermacoceae. This requires a morphological investigation
across taxa to find character support for the many new
phylogenetic relationships detected.
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Posada, D. & K. A. Crandall. 1998. Modeltest: Testing the
model of DNA substitution. Bioinformatics 14: 817–818.
Puff, C. 1986. Phylohydrax (Rubiaceae–Spermacoceae)—A
new genus to accommodate the African and Madagascan
Hydrophylax species. Pl. Syst. Evol. 154: 343–366.
Robbrecht, E. 1988. Tropical woody Rubiaceae. Characteristic features and progressions. Contributions to a new
subfamilial classification. Opera Bot. Belg. 1: 1–271.
———. 1993. Supplement to the 1988 outline of the
classification of the Rubiaceae Index to genera. In E.
Robbrecht (editor), Advances in Rubiaceae Macrosystematics. Opera Bot. Belg. 6: 173–196.
——— & J. F. Manen. 2006. The major evolutionary lineages
of the coffee family (Rubiaceae, angiosperms). Combined
analysis (nDNA and cpDNA) to infer the position of
Coptosapelta and Luculia, and supertree construction based
on rbcL, rps16, trnL-trnF and atpB-rbcL data. A new
classification in two subfamilies, Cinchonoideae and
Rubioideae. Syst. Geogr. Pl. 76: 85–146.
Rogers, G. K. 1987. The genera of Cinchonoideae (Rubiaceae) in the southeastern United States. J. Arnold Arbor.
68: 137–183.
Ronquist, F. & J. P. Huelsenbeck. 2003. MRBAYES 3:
Bayesian phylogenetic inference under mixed models.
Bioinformatics 19: 1572–1574.
Scheltens, A. 1998. Pollenmorfologische studie van de
Afrikaanse Hedyotideae (Rubiaceae). Licentiate Thesis,
Katholieke Universiteit Leuven, Leuven, Belgium.
Schumann, K. 1891. Rubiaceae. In A. Engler & K. Prantl
(editors), Die natürlichen Pflanzenfamilien 4: 1–156.
Simmons, M. P. & H. Ochoterena. 2000. Gaps as characters
in sequence-based phylogenetic analyses. Syst. Biol. 49:
369–381.
Staden, R., K. Beal & J. Bonfield. 1998. The Staden Package.
Pp. 115–130 in S. Miseners & S. Krawetz (editors),
Computer Methods in Molecular Biology. The Humana
Press Inc., New York.
Suzuki, Y., G. V. Glazko & M. Nei. 2002. Over credibility of
molecular phylogenies obtained by Bayesian phylogenetics. Proc. Natl. Acad. Sci. U.S.A. 99: 16138–16143.
Swofford, D. 2002. PAUP*: Phylogenetic Analysis Using
Parsimony (* and Other Methods), Vers. 4. Sinauer
Associates, Sunderland, Massachusetts.
Taberlet, P., G. Gielly, G. Pautou & J. Bouvet. 1991.
Universal primers for amplification of three noncoding regions of chloroplast DNA. Pl. Mol. Biol. 17:
1105–1109.
Terrell, E. E. 1975. Relationships of Hedyotis fruticosa L. to
Houstonia L. and Oldenlandia L. Phytologia 31: 418–421.
———. 1987. Carterella (Rubiaceae), new genus from Baja
California, Mexico. Brittonia 39: 248–252.
———. 1990. Synopsis of Oldenlandia (Rubiaceae) in the
United States. Phytologia 68: 125–133.
———. 1991. Overview and annotated list of North
American species of Hedyotis, Houstonia, Oldenlandia
(Rubiaceae), and related genera. Phytologia 71: 212–243.
———. 1996. Revision of Houstonia (Rubiaceae–Hedyotideae). Syst. Bot. Monogr. 48: 1–118.
———. 2001a. Taxonomy of Stenaria (Rubiaceae; Hedyotideae), a new genus including Hedyotis nigricans. Sida 19:
591–614.
———. 2001b. Stenotis (Rubiaceae), a new segregate genus
from Baja California, Mexico. Sida 19: 899–911.
———. 2001c. Taxonomic review of Houstonia acerosa and
H. palmeri, with notes on Hedyotis and Oldenlandia
(Rubiaceae). Sida 19: 913–922.
——— & W. H. Lewis. 1990. Oldenlandiopsis (Rubiaceae),
a new genus from the Caribbean basin, based on
Oldenlandia callitrichoides Grisebach. Brittonia 42:
185–190.
——— & H. Robinson. 2003. Survey of Asian and Pacific
species of Hedyotis and Exallage (Rubiaceae) with
nomenclatural notes on Hedyotis types. Taxon 52:
775–782.
———, W. H. Lewis, H. Robinson & J. W. Nowicke. 1986.
Phylogenetic implications of diverse seed types, chromosome numbers, and pollen morphology in Houstonia
(Rubiaceae). Amer. J. Bot. 73: 103–115.
———, H. E. Robinson, W. L. Wagner & D. H. Lorence.
2005. Resurrection of genus Kadua for Hawaiian
Hedyotidinae (Rubiaceae), with emphasis on seed and
fruit characters and notes on South Pacific species. Syst.
Bot. 30: 818–833.
Thompson, J. D., T. J. Gibson, F. Plewniak, F. Jeanmougin &
D. G. Higgins. 1997. The ClustalX windows interface:
Flexible strategies for multiple sequence alignment aided
by quality analyses tools. Nucl. Acids Res. 25: 4876–
4882.
Thulin, M. & B. Bremer. 2004. Studies in the tribe
Spermacoceae (Rubiaceae–Rubioideae): The circumscriptions of Amphiasma and Pentanopsis and the affinities of
Phylohydrax. Pl. Syst. Evol. 247: 233–239.
Verdcourt, B. 1958. Remarks on the classification of
the Rubiaceae. Bull. Jard. Bot. État Bruxelles 28:
209–281.
———. 1976. Rubiaceae (part 1). Pp. 1–414 in R. M. Polhill
(editor), Flora of Tropical East Africa. Crown Agents for
Overseas Governments and Administrations, London.
Wagner, W. L., D. R. Herbst & S. H. Sohmer. 1989.
Contributions to the flora of Hawaii: 2. Begoniaceae:
Violaceae and the monocotyledons. Bishop Mus. Occas.
Pap. 29: 88–130.
Taxon
Conostomium (Stapf) Cufod.
C. natalense (Hochst.) Bremek.
C. quadrangulare (Rendle) Cufod.
C. zoutpansbergense (Bremek.) Bremek.
Crusea Cham. & Schltdl.
C. calocephala DC.
C. megalocarpa (A. Gray) S. Watson
Dentella J. R. Forst. & G. Forst.
D. dioeca Airy Shaw
atpB-rbcL
rps16 intron
trnL-trnF
Zambia, Dessein et al. 671 (BR)
Zambia, Dessein et al. 201 (BR)
EU542917*
EU542918*
EU543018*
EU543019*
EU543077*
EU543078*
Angola, Kers 3350 (S)
Zambia, Dessein et al. 1167 (BR)
EU542919*
EU542920*
AF002753(1)
EU543020*
EU543079*
EU543080*
Ecuador, Hekker & Hekking 10335 (GB)
Ecuador, Øllgaard et al. 58395 (NY)
unknown, Edwin et al. 3624 (S)
Costa Rica, Cronquist 8827 (NY)
Peru, Wurdack 1073 (NY)
Colombia, Andersson et al. 2195 (GB)
Venezuela, Pipoly et al. 6467 (GB)
Ecuador, Harling & Andersson 22232 (GB)
Mexico, Stafford et al. 203 (MO)
Colombia, Andersson et al. 2196 (GB)
Ecuador, Ståhl 4481 (GB)
–
–
–
–
–
EU542921*
–
EU542922*
–
–
EU542923*
AF333348(2)
AF333350(2)
AF333352(2)
AF333354(2)
AF333356(2)
AF002754(1)
AF333359(2)
AF333362(2)
AF333364(2)
AF002755(1)
AF333366(2)
AF333349(2)
AF333351(2)
AF333353(2)
AF333355(2)
AF333357(2)
EU543081*
–
AF333363(2)
–
AF333365(2)
EU543082*
cult., Forbes s.n. (S)
unknown, Van Caekenberghe 264 (cult.
at BR)
Mexico, Spencer et al. 363 (NY)
EU542925*
–
EU543022*
AF002758(1)
EU543084*
–
–
–
EU642537*
South Africa, Dahlstrand 1346 (GB)
South Africa, Bremer et al. 4341 (UPS)
Ethiopia, Puff & Kelbessa 821222 (UPS)
South Africa, Bremer et al. 4331 (UPS)
–
EU542927*
EU542928*
EU542929*
AF002760(1)
–
EU543024*
–
EU543085*
–
EU543086*
EU543087*
Guatemala, Gustafsson et al. 215 (GB)
Mexico, Pringle 3852 (S)
EU542930*
EU542931*
–
EU543025*
EU543088*
EU543089*
–
–
EU543090*
Voucher information
Australia, Harwood 1559 (BR)
Annals of the
Missouri Botanical Garden
Agathisanthemum Klotzsch
A. bojeri Klotzsch
A. globosum (Hochst. ex A. Rich.) Klotzsch
Amphiasma Bremek.
A. benguellense (Hiern) Bremek.
A. luzuloides (K. Schum.) Bremek.
Arcytophyllum Willd. ex Schult. & Schult. f.
A. aristatum Standl.
A. ciliolatum Standl.
A. ericoides (Willd. ex Roem. & Schult.) Standl.
A. lavarum K. Schum.
A. macbridei Standl.
A. muticum (Wedd.) Standl.
A. nitidum (Kunth) Schltdl.
A. rivetii Danguy & Cherm.
A. serpyllaceum (Schltdl.) Terrell
A. setosum (Ruiz & Pav.) Schltdl.
A. thymifolium (Ruiz & Pav.) Standl.
Bouvardia Salisb.
B. glaberrima Engelm.
B. ternifolia (Cav.) Schltdl.
128
Appendix 1. List of taxa used in the phylogenetic analyses with voucher information (geographic origin, collector, collector number, herbarium), accession numbers, and literature citations
from previously published sequences for the three plastid markers atpB-rbcL, rps16 intron, and trnL-trnF: (1) Andersson & Rova, 1999; (2) Andersson et al., 2002; (3) Dessein et al., 2005a. New
sequences are marked with an asterisk. Missing sequences are marked with a dash.
Taxon
rps16 intron
trnL-trnF
EU542932*
AF333370(2)
EU543091*
Kenya, Strid 2598 (GB)
EU542933*
AF002761(1)
–
Kenya, Luke 9029 (UPS)
French Guiana, Anderson et al. 2071 (GB)
French Guiana, Anderson et al. 1961 (GB)
EU542934*
–
EU542935*
EU543026*
AF002762(1)
EU543027*
EU543092*
–
EU543093*
Trinidad, Hummel s.n. (GB)
EU542936*
AY764289(3)
EU543094*
Cuba, Rova et al. 2286 (GB)
EU542937*
AF002763(1)
EU543095*
Argentina, Vanni & Radovancick 996 (GB)
Argentina, Schinini & Cristobal 9811 (GB)
EU542938*
EU542939*
AY764290(3)
EU543028*
EU543096*
EU543097*
–
AY764291(3)
–
Hong Kong, Shiu Ying Hu 10821 (S)
Sri Lanka, Larsson & Pyddoke 22 (S)
Caroline Islands, Fosberg 47697 (S)
Sri Lanka, Wambeek & Wanntorp 2996 (S)
Sri Lanka, Klackenberg 413 (S)
Sri Lanka, Fagerlind 3668 (S)
Malaysia, Sabah, Wallander 6 (GB)
Sri Lanka, Bremer et al. 163 (S)
Sri Lanka, Fagerlind 5082 (S)
South India, Klackenberg & Lundin 03 (S)
EU542941*
EU542942*
EU542943*
EU542944*
EU542945*
EU542946*
EU542947*
EU542948*
EU542949*
EU542950*
–
–
–
–
EU543029*
EU543030*
AF002767(1)
–
–
EU543031*
–
EU543098*
EU543099*
–
EU543100*
EU543101*
EU543102*
EU543103*
EU543104*
EU543105*
Zambia, Dessein et al. 1017 (BR)
EU542951*
EU543032*
EU543107*
French Guiana, Andersson et al. 2040 (GB)
EU542952*
–
EU543108*
U.S.A., Vincent & Lammers s.n. (GB)
U.S.A., Yatskievych 96-49 (MO)
U.S.A., Weigend 9963 (NY)
EU542953*
EU542954*
–
AF333379(2)
AF002766(1)
–
EU543109*
–
EU642536*
Voucher information
Madagascar, De Block et al. 569 (BR)
129
atpB-rbcL
Australia, Andersson 2262 (GB)
Groeninckx et al.
Phylogeny of Spermacoceae
D. repens (L.) J. R. Forst. & G. Forst.
Dibrachionostylus Bremek.
D. kaessneri (S. Moore) Bremek.
Diodia L. as traditionally delimited
D. aulacosperma K. Schum.
D. sarmentosa Sw.
D. spicata Miq.
Emmeorhiza Pohl ex Endl.
E. umbellata (Spreng.) K. Schum.
Ernodea Sw.
E. littoralis Sw.
Galianthe Griseb.
G. brasiliensis (Spreng.) E. L. Cabral & Bacigalupo
G. eupatorioides (Cham. & Schltdl.) E. L. Cabral
Gomphocalyx Baker
G. herniarioides Baker
Hedyotis L.
H. consanguinea Hance
H. fruticosa L.
H. korrorensis (Valeton) Hosok.
H. lawsoniae Wight
H. lessertiana Arn. var. lassertiana Thwaites
H. lessertiana var. marginata Thwaites & Trimen
H. macrostegia Stapf
H. quinquinervis Thwaites
H. rhinophylla Thwaites ex Trimen
H. swertioides Hook. f.
Hedythyrsus Bremek.
H. spermacocinus (K. Schum.) Bremek.
Hemidiodia K. Schum.
H. ocymifolia (Willd. ex Roem. & Schult.) K. Schum.
Houstonia L.
H. caerulea L.
H. longifolia Gaertn.
Volume 96, Number 1
2009
Appendix 1. Continued.
130
Appendix 1. Continued.
Taxon
Kadua Cham. & Schltdl.
K. acuminata Cham. & Schltdl.
K. affinis Cham. & Schltdl.
K. axillaris (Wawra) W. L. Wagner & Lorence
K.
K.
K.
K.
K.
K.
K.
atpB-rbcL
rps16 intron
trnL-trnF
U.S.A., Hawaii, cult. at BR
U.S.A., Hawaii, Motley 1733 (NY)
U.S.A., Hawaii, Harrison-Gagne s.n. (GB)
U.S.A., Hawaii, Maui, Motley 1724 (NY)
U.S.A., Hawaii, Skottsberg 6788 (S)
cult., Lorence 8021 (PTBG)
U.S.A., Hawaii, Motley 1703 (NY)
cult., Wood 5062 (PTGB)
U.S.A., Hawaii, Kauai, Wagner 6350 (BISH)
U.S.A., Hawaii, Oahu, Motley 1747 (NY)
U.S.A., Hawaii, Kauai, Perlman 15631 (BISH)
EU542955*
–
–
–
EU542956*
EU542957*
–
EU542958*
–
–
–
–
EU642523*
AF002765(1)
EU642524*
EU543033*
AF333376(2)
EU642525*
AF333371(2)
EU642526*
EU642527*
EU642528*
EU543110*
EU642538*
–
EU642535*
EU543111*
EU543112*
EU642539*
EU543113*
EU642540*
EU642541*
EU642542*
U.S.A., Hawaii, Sparre 27 (S)
U.S.A., Hawaii, Oahu, Motley 1677 (NY)
EU542959*
–
–
EU642529*
EU543114*
EU642543*
U.S.A., Hawaii, Molokai, Perlman 6647 (BISH)
U.S.A., Hawaii, Molokai, Kiehn & Luegmayr 920823 (WU)
cult., Perlman 12783 (GB)
Rapa Island, French Polynesia, Perlman 17953 (NY)
–
EU542960*
EU542961*
–
EU642530*
EU543034*
AF333375(2)
EU642531*
EU642544*
EU543115*
EU543116*
EU642545*
South Africa, Bremer et al. 4307 (UPS)
Zambia, Dessein et al. 432 (BR)
Zambia, Dessein et al. 751 (BR)
South Africa, Dessein et al. 469 (BR)
Zambia, Dessein et al. 1149 (BR)
Kenya, Luke 9035 (UPS)
Burkina Faso, Madsen 5940 (NY)
Zambia, Dessein et al. 470 (BR)
Madagascar, De Block et al. 539 (BR)
EU542962*
EU542963*
EU542964*
EU542965*
EU542966*
EU542967*
–
EU542968*
EU542969*
EU543035*
EU543036*
EU543037*
EU543038*
EU543039*
EU543040*
–
EU543041*
–
EU543117*
EU543118*
EU543119*
EU543120*
EU543121*
EU543122*
EU642546*
EU543123*
EU543124*
Tanzania, Gereau 2513 (BR)
EU542970*
–
EU543125*
French Guiana, Andersson et al. 1917 (GB)
Colombia, Andersson et al. 2128 (GB)
EU542971*
EU542972*
AF002768(1)
AF002769(1)
–
EU543126*
Annals of the
Missouri Botanical Garden
centranthoides Hook. & Arn.
cordata Cham. & Schltdl.
coriacea (J. E. Smith) W. L. Wagner & Lorence
degeneri (Fosberg) W. L. Wagner & Lorence
elatior (H. Mann) W. L. Wagner & Lorence
fluviatilis C. N. Forbes
flynnii (W. L. Wagner & Lorence) W. L. Wagner
& Lorence
K. foggiana (Fosberg) W. L. Wagner & Lorence
K. fosbergii (W. L. Wagner & D. R. Herbst) W. L.
Wagner & Lorence
K. laxiflora H. Mann
K. littoralis Hillebr.
K. parvula A. Gray
K. rapensis F. Br.
Kohautia Cham. & Schltdl.
K. amatymbica Eckl. & Zeyh.
K. caespitosa Schnizl.
K. coccinea Royle
K. cynanchica DC.
K. microcala Bremek.
K. obtusiloba (Hiern) Bremek.
K. senegalensis Cham. & Schltdl.
K. subverticillata (K. Schum.) D. Mantell
K. virgata (Willd.) Bremek.
Lelya Bremek.
L. osteocarpa Bremek.
Manettia Mutis ex L.
M. alba (Aubl.) Wernham
M. lygistum (L.) Sw.
Voucher information
Taxon
rps16 intron
trnL-trnF
Zambia, Dessein et al. 265 (BR)
EU542973*
EU543042*
EU543127*
French Guiana, Andersson et al. 1995 (GB)
Guiana, Jansen-Jacobs et al. 4785 (GB)
EU542974*
EU542975*
AF002770(1)
EU543044*
EU543128*
–
Zambia, Dessein et al. 1273 (BR)
EU542976*
EU543045*
EU543129*
–
AF003607(1)
–
EU542977*
EU542978*
EU542979*
EU542980*
EU542981*
EU542982*
EU542983*
EU542984*
EU542985*
EU542986*
EU542987*
EU542988*
EU542989*
EU542990*
EU542991*
EU542992*
EU542993*
EU542994*
–
–
–
EU542995*
EU542996*
EU542997*
EU542998*
EU543046*
EU543047*
–
EU543048*
EU543049*
EU543050*
EU543051*
–
EU543052*
EU543053*
EU543054*
EU543055*
EU543056*
EU543057*
EU543058*
EU543059*
–
EU543060*
AF333382(2)
EU543061*
EU543043*
AY764294(3)
–
EU543062*
AY764293(3)
EU543130*
EU543131*
EU543132*
EU543133*
EU543134*
EU543135*
EU543136*
EU543137*
EU543138*
EU543139*
EU543140*
EU543141*
EU543142*
EU543143*
EU543144*
EU543145*
EU543146*
–
–
EU543147*
–
EU543148*
EU543149*
EU543106*
–
cult., Chase 2915 (K)
Zambia, Dessein et al. 627 (BR)
Zambia, Dessein et al. 932 (BR)
Japan, Van Caekenberghe 63 (cult. at BR)
Zambia, Dessein et al. 843 (BR)
Tanzania, Kayombo et al. s.n. (BR)
Zambia, Dessein et al. 487 (BR)
Zambia, Dessein et al. 928 (BR)
Tanzania, Kayombo & Kahemela 1993 (BR)
Zambia, Dessein et al. 1019 (BR)
Australia, Harwood 1511 (BR)
Zambia, Dessein et al. 935 (BR)
Zambia, Dessein et al. 1286 (BR)
Zambia, Dessein et al. 442 (BR)
Zambia, Dessein et al. 463 (BR)
Zambia, Dessein et al. 1356 (BR)
Mexico, Frödeström & Hultén 681 (S)
Australia, Harwood 1516 (BR)
Zambia, Dessein et al. 924 (BR)
Gabon, Andersson & Nilsson 2326 (GB)
Zambia, Dessein et al. 346 (BR)
Zambia, Dessein et al. 1197 (BR)
Brazil, Harley 15514 (UPS)
Tanzania, Bidgood et al. 4015 (BR)
Japan, Van Caekenberghe 70 (cult. at BR)
Guyana, Jansen-Jacobs et al. 41 (UPS)
131
atpB-rbcL
Voucher information
Groeninckx et al.
Phylogeny of Spermacoceae
Manostachya Bremek.
M. ternifolia E. S. Martins
Mitracarpus Zucc. ex Schult. & Schult. f.
M. frigidus (Willd. ex Roem. & Schult.) K. Schum.
M. microspermus K. Schum.
Mitrasacmopsis Jovet
M. quadrivalvis Jovet
Nesohedyotis (Hook. f.) Bremek.
N. arborea (Roxb.) Bremek.
Oldenlandia L.
O. affinis (Roem. & Schult.) DC.
O. angolensis K. Schum.
O. biflora L.
O. capensis L. f. var. capensis
O. capensis var. pleiosepala Bremek.
O. corymbosa L.
O. echinulosa K. Schum.
O. echinulosa K. Schum. var. pellucida (Hiern) Verdc.
O. fastigiata Bremek.
O. galioides (F. Muell.) F. Muell.
O. geophila Bremek.
O. goreensis (DC.) Summerh.
O. herbacea (L.) Roxb. var. goetzei Bremek.
O. herbacea (L.) Roxb. var. herbacea
O. lancifolia (Schumach.) DC.
O. microtheca (Cham. & Schltdl.) DC.
O. mitrasacmoides F. Muell.
O. nematocaulis Bremek.
O. nervosa Hiern
O. robinsonii Pit.
O. rosulata K. Schum.
O. salzmannii (DC.) Benth. & Hook. f. ex B. D. Jacks.
O. taborensis Bremek.
O. tenelliflora (Blume) Kuntze
O. tenuis K. Schum.
Volume 96, Number 1
2009
Appendix 1. Continued.
132
Appendix 1. Continued.
Taxon
U.S.A., Godfrey 57268 (GB)
Ethiopia, Friis et al. 2560 (UPS)
Kenya, Luke & Luke 8362 (UPS)
atpB-rbcL
rps16 intron
trnL-trnF
EU542999*
EU543017*
EU543000*
(3)
AY764295
EU543076*
EU543063*
EU543150*
EU543168*
EU543151*
–
EU543065*
EU543153*
Zambia, Dessein et al. 598 (BR)
EU543002*
EU543066*
EU543154*
South Africa, Bremer 3783 (UPS)
Madagascar, De Block et al. 640 (BR)
EU543003*
EU543004*
EU543067*
AY764292(3)
–
EU543155*
Colombia, Andersson et al. 2073 (GB)
Australia, Egerod 85343 (GB)
EU543005*
EU543006*
AF003614(1)
EU543068*
EU543156*
EU543157*
French Guiana, Andersson 1908 (GB)
Colombia, Andersson et al. 2074 (GB)
Australia, Harwood 1148 (BR)
Madagascar, De Block et al. 794 (BR)
Kenya, Luke 9022 (UPS)
Sri Lanka, Wanntorp et al. 2667 (S)
Colombia, Andersson et al. 2078 (GB)
French Guiana, Andersson et al. 2016 (GB)
Gabon, Andersson & Nilsson 2296 (GB)
EU543007*
–
EU543008*
EU543010*
EU543009*
EU543011*
EU543012*
EU543013*
EU543014*
EU543069*
AF003619(1)
EU543070*
EU543072*
EU543071*
EU543073*
–
–
EU543074*
EU543158*
–
EU543159*
EU543161*
EU543160*
EU543162*
EU543163*
EU543164*
EU543165*
U.S.A., Yatskievych 96-92 (MO)
EU543015*
AF333373(2)
EU543166*
Australia, Lazarides & Palmer 272 (K)
EU543016*
EU543075*
EU543167*
Zambia, Dessein et al. 264 (BR)
EU542924*
EU543021*
EU543083*
Madagascar, De Block et al. 578 (BR)
EU542926*
EU543023*
–
Zambia, Dessein et al. 678 (BR)
EU543001*
EU543064*
EU543152*
Ethiopia, Gilbert et al. 7458 (UPS)
Annals of the
Missouri Botanical Garden
O. uniflora L.
O. wauensis Schweinf. ex Hiern
O. wiedemannii K. Schum.
Pentanopsis Rendle
P. fragrans Rendle
Pentodon Hochst.
P. pentandrus (K. Schum. & Thonn.) Vatke
Phylohydrax Puff
P. carnosa (Hochst.) Puff
P. madagascariensis (Willd. ex Roem. & Schult.) Puff
Richardia L.
R. scabra L.
R. stellaris (Cham. & Schltdl.) Steud.
Spermacoce L.
S. capitata Ruiz & Pav.
S. confusa Rendle ex Gillis
S. erosa Harwood
S. flagelliformis Poir.
S. filituba (K. Schum.) Verdc.
S. hispida L.
S. prostrata Aubl.
S. remota Lam.
S. ruelliae DC.
Stenaria (Raf.) Terrell
S. nigricans (Lam.) Terrell
Synaptantha Hook. f.
S. tillaeacea (F. Muell.) Hook. f.
OUTGROUP TAXA
Batopedina Verdc.
B. pulvinellata Robbr.
Carphalea Juss.
C. madagascariensis Lam.
Pentanisia Harv.
P. parviflora Stapf ex Verdc.
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