Phylogeny of Chamaecrista ser.

Coriaceae (Leguminosae) Unveils a Lineage Recently Diversified

in Brazilian Campo Rupestre Vegetation
Author(s): Juliana Gastaldello Rando, Alexandre R. Zuntini, Adilva Souza Conceição, Cássio van
den Berg, José Rubens Pirani and Luciano Paganucci de Queiroz,
Source: International Journal of Plant Sciences, (-Not available-), p. 000
Published by: The University of Chicago Press
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Int. J. Plant Sci. 177(1):000–000. 2016. SYMPOSIUM PAPER
q 2015 by The University of Chicago. All rights reserved.
1058-5893/2016/17701-00XX$15.00 DOI: 10.1086/683846

PHYLOGENY OF CHAMAECRISTA SER. CORIACEAE (LEGUMINOSAE) UNVEILS A LINEAGE

RECENTLY DIVERSIFIED IN BRAZILIAN CAMPO RUPESTRE VEGETATION
Juliana Gastaldello Rando,1,* Alexandre R. Zuntini,* Adilva Souza Conceição,† Cássio van den Berg,‡
José Rubens Pirani,* and Luciano Paganucci de Queiroz‡
*Departamento de Botânica, Instituto de Biociências, Universidade de São Paulo, Rua do Matão 277, 05508-900, São Paulo, São Paulo, Brazil;
†Colegiado de Ciências, Universidade do Estado da Bahia, Campus VIII, Rua do Gangorra, 503, CHESF, Paulo Afonso, Bahia, 48.608-290,
Brazil; and ‡Laboratório de Sistemática Molecular de Plantas, Departamento de Ciências Biológicas, Universidade Estadual de Feira
de Santana, Km 03 BR 116, Campus, 44031-460, Feira de Santana, Bahia, Brazil

Editors: Gwilym P. Lewis and Patrick S. Herendeen

Premise of research. Chamaecrista sect. Chamaecrista ser. Coriaceae comprises 22 species, mostly distrib-
uted in the highlands of Brazil in Cerrado (Neotropical savanna) and Campo Rupestre (upland rocky field) veg-
etation. Phylogenetic studies for plant taxa of these rich and endemic Brazilian floras are still scarce. In this con-
text, the main objective of our study was to examine the phylogenetic history of a plant group diversified in the
mountainous areas of central Brazil.
Methodology. The phylogeny of Chamaecrista ser. Coriaceae was examined with combined data from
two nuclear regions (ETS [external transcribed spacer] and ITS), two plastid regions (trnD-T and trnL-F),
and morphological evidence from comprehensive sampling of the series. We used phylogenetic analyses to
evaluate morphological-character evolution to identify putative synapomorphies, and we used Bayesian esti-
mation to explore the timing of diversification of the group.
Pivotal results. The series, as currently circumscribed, appears as polyphyletic in all of the analyses, but
most of the species ascribed to it form a well-supported clade endemic to highland areas in the Brazilian Cer-
rado, referred to as the Coriaceae clade. The plastid regions provided high support for the monophyly of this
clade, with the nuclear regions providing better resolution within the clade. In addition to molecular evidence,
two putative morphological synapomorphies were found. The age-estimation analyses show a recent diversi-
fication in the group.
Conclusions. Molecular and morphological data indicated that a narrower definition of Chamaecrista ser.
Coriaceae as a monophyletic group is required. In addition to two clear deletions in the plastid sequences stud-
ied, the presence of a woody underground system and flowers with one stamen displaced to the same side as
the gynoecium appear as putative morphological synapomorphies. The low resolution found among species
within the clade may reflect a recent radiation that has occurred in the past 5 Myr—a common pattern in other
groups restricted to highland areas in South America.

Keywords: Brazil, Campo Rupestre vegetation, Cerrado, endemism, recent diversification, taxonomy.

Online enhancements: appendix tables.

Introduction vegetation of the Espinhaço Range in the states of Minas Gerais

Chamaecrista Moench is one of the richest genera of flower-
ing plants in the Brazilian savannas, with its greatest diversity
occurring in the Cerrado domain on the central Brazilian plateau
and in upland areas in Campo Rupestre (literally “rocky field”)
1
Author for correspondence. Present address: Programa de Pós-
Graduação em Ciências Ambientais, Universidade Federal do Oeste
da Bahia, Rua Professor José Seabra de Lemos 316, Barreiras, Bahia,
47.808-021, Brazil; e-mail: ju_rando@hotmail.com.
Manuscript received May 2015; revised manuscript received July 2015; elec-
tronically published December 1, 2015.
and Bahia (Irwin and Barneby 1982; Lewis 1987; Rando and
Pirani 2011; Souza and Bortoluzzi 2015). The genus comprises
more than 330 species distributed mainly in tropical America,
with few species in Africa, Asia, and Australia, occasionally ex-
tending to temperate areas (Irwin and Barneby 1982; Lewis
2005). In Brazil, 253 species of Chamaecrista occur, 204 of them
endemic, and many have narrow distribution ranges (Rando
and Pirani 2011; Souza and Bortoluzzi 2015).
Phylogenetic studies based on molecular markers support
the monophyly of the genus (Conceição et al. 2009). Irwin and
Barneby (1978, 1982) recognized six sections in the genus. Sec-
tion Chamaecrista is the second-largest section, including ca. 76

000

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000 INTERNATIONAL JOURNAL OF PLANT SCIENCES

species globally and ca. 52 species in the Americas, and is di-
vided into six series based on the position of the inflorescence,
the number of stamens, patterns of leaflet venation, and the pres-
ence of a xylopodium (Irwin and Barneby 1982). Chamaecrista
sect. Chamaecrista ser. Coriaceae (Benth.) H.S. Irwin and Bar-
neby, the focus of this study, embraces 22 species and six varie-
ties (Irwin and Barneby 1982; Rando et al. 2013) and is charac-
terized by a shrubby habit with twigs sprouting from a woody
rootstock (a xylopodium, according to Irwin and Barneby 1982;
fig. 1D), extrafloral nectaries on the petiole, thick-textured leaflets,

Fig. 1 A, Map of part of South America and the Antilles depicting the distribution of Chamaecrista ser. Coriaceae according to Irwin and
Barneby (1982). Darkening shades of brown indicate increasing elevations. The Espinhaço Range is delimited by the rhombic area. The following
Brazilian states are highlighted: Bahia (BA), Espírito Santo (ES), Goiás (GO), Minas Gerais (MG), Paraná (PR), Roraima (RR), and São Paulo
(SP). B, Typical landscape of Campo Rupestre vegetation, illustrating its predominantly herbaceous layer among rocky outcrops. C, Habit
of Chamaecrista rossicorum. D, E, Chamaecrista papillata after the passage of fire, showing dead, burned branches and new green sprouts
(D) and detail of a woody rootstock from which the plants resprout (E).

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 RANDO ET AL.—PHYLOGENY OF CHAMAECRISTA SER. CORIACEAE
and axillary peduncles. Although the series was supported as
monophyletic in a previous phylogenetic analysis (Conceição
et al. 2009), that study included only four species, and therefore
the monophyly remains to be confirmed with a more compre-
hensive sampling.
As currently circumscribed (Irwin and Barneby 1982; Rando
et al. 2013), 19 of the 22 species are restricted to the moun-
tains of the Espinhaço Range, occurring in areas of Campo
Rupestre vegetation. In addition, one coastal endemic species,
Chamaecrista caribaea (Northr.) Britton, is found in the Ba-
hamas (fig. 1A), one species is restricted to the state of Goiás, cen-
tral Brazil, and another is disjunctly distributed in elevated areas
of the tepuis in Roraima (northern Brazil) and Bahia (eastern
Brazil; Irwin and Barneby 1982; Rando and Pirani 2011).
Campo Rupestre (fig. 1B, 1C) is the dominant vegetation type
of the Espinhaço Range, harboring high diversity in several plant
groups and many endemic species (Giulietti et al. 1997; Rapini
et al. 2008; Bitencourt and Rapini 2013; Ribeiro et al. 2014).
Campo Rupestre areas are generally separated one from an-
other and have been treated as isolated islands surrounded by
either Caatinga (seasonally dry tropical forests and woodlands)
or Cerrado vegetation (Prance 1994; Pennington et al. 2000,
2006; Fiaschi and Pirani 2009). Phylogenetic studies of the plant
taxa occurring in this rich and endemic flora have been scarce,
but they suggest a recent radiation (Quaternary) of plant species
throughout the Cerrado and Campo Rupestre, with high species
turnover (Fiaschi and Pirani 2009; Queiroz et al. 2009; Simon
et al. 2009; Koenen et al. 2013). Similar morphological traits
occur convergently in several independent plant lineages in Cer-
rado and Campo Rupestre vegetations, suggesting that they are
adaptations that allow plant survival in dry habitats with fre-
quent fire regimes (Rizzini and Heringer 1961; Simon et al.
2009; Simon and Pennington 2012). Some of these traits are
typical of Chamaecrista ser. Coriaceae species, including leaf-
lets incrassate and indurate in texture and the presence of a
woody rootstock (fig. 1D, 1E), making Chamaecrista sect.
Coriaceae an interesting group to study the evolution of Cer-
rado and Campo Rupestre floras.
Taking into consideration the lack of a densely sampled phy-
logenetic evaluation of series Coriaceae, its strong ecological ties
with Campo Rupestre vegetation, and its wide geographical dis-
junction between central-eastern Brazil and the Caribbean, we
sought to develop a robust phylogenetic framework of the series
to (1) test the monophyly of series Coriaceae, (2) assess the phylo-
genetic position of the series within the genus, (3) characterize its
morphological evolution, and (4) explore the timing of diversifi-
cation of the series in Campo Rupestre vegetation.
Material and Methods
Taxon Sampling and Molecular and Morphological Data
Two separate analyses were performed, using two sampling
strategies. “Broad sampling” was designed to test the mono-
phyly of Chamaecrista ser. Coriaceae and the phylogenetic po-
sition within the genus, including representatives of all of the
sections and series recognized in Chamaecrista that were ex-
amined in a previous phylogenetic study of the genus (Con-
ceição et al. 2009). This set of analyses included 78 vouchered
sequences of the plastid trnL-F and internal transcribed spacer
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000
(ITS) regions, including 56 species (74 taxa) of Chamaecrista
(38 from GenBank and 36 new sequences) and four representatives
of the genera Cassia and Senna as outgroups (Bruneau et al.
2008). “Narrow sampling” was used to assess phylogenetic
relationships among species within Chamaecrista ser. Coria-
ceae. We included 46 accessions (21 species of Chamaecrista
ser. Coriaceae and 10 outgroup taxa) for the plastid trnL-F re-
gion, 49 accessions (21 species of Chamaecrista ser. Coriaceae
and 9 outgroup taxa) for the plastid trnD-T, 35 accessions (20
species of Chamaecrista ser. Coriaceae and 9 as outgroup) for
the nuclear ITS, and 39 accessions (20 species of Chamaecrista
ser. Coriaceae and 8 as outgroup) for the nuclear external tran-
scribed spacer (ETS). We included 10 species from all sections
of Chamaecrista as outgroups (app. A; table 1). In the combined
analysis, we included 46 accessions (21 species of Chamaecrista
ser. Coriaceae and 10 as outgroup). Chamaecrista multinervia
(Mart. ex Benth.) H.S. Irwin & Barneby was the only species of
Chamaecrista ser. Coriaceae not examined in this study.
Morphological characters were surveyed for all of the taxa
in the narrow-sampling study. They were coded from herbar-
ium sheets, liquid-preserved samples, and living plants in the
field. All nomenclatural types were analyzed. Only fully de-
veloped structures of mature plants (reproductive individuals)
were considered. Specimens surveyed were from the following
herbaria: ALCB, B, BHCB, DIAM, EAC, ESA, F, G, GH,
HRCB, HUEFS, HUFU, INPA, K, LE, M, MBM, MG, MIRR,
NY, OUPR, P, R, RB, SP, SPF, UB, UEC, UFG, and US (ac-
ronyms according to Thiers 2015). On the basis of our de-
tailed morphological investigations, 27 variable characters were
identified and coded as binary or as unordered multistate char-
acters. The morphological terminology and characters and char-
acter states used are detailed in appendix B. The morphological
data matrix is available in appendix C (apps. B, C available
online).
DNA Extraction, Amplification, and Sequencing
Total DNA was extracted, mostly from fresh or silica gel–
dried leaflets, with an adaptation of the 2# CTAB (cetyltrimeth-
ylammonium bromide) procedure of Doyle and Doyle (1987)
or an Invitek mini plant kit (Invitek, Berlin). Variable molecular
regions were chosen on the basis of a pilot study we performed
that sampled species from series Coriaceae and other sections
and series of Chamaecrista. The chloroplast DNA (cpDNA) rpl32-
trnL spacer and rps16 intron regions were tested and subse-
quently discarded because of their low species-level resolution.
The trnD-T, trnL-F, ETS, and ITS 5.8S regions appeared suffi-
ciently variable at the species level and were selected for this study.
Two universal primers (C and F) of Taberlet et al. (1991) were
used for the amplification and sequencing of trnL-F. The prim-
ers trnD (F) and trnT (R) were used for the amplification and se-
quencing of trnD-T (Shaw et al. 2005). Amplifications of both
plastid regions were performed with Toptaq Master Mix (Qia-
gen, Dusseldorf, Germany) scaled to 15 mL, containing 7.5 mL
premade mix, 0.2 mL of each primer (15 mM), 6.6 mL of H2O
(Qiagen), and 0.5 mL of DNA (20–40 ng). The program for
trnL-F consisted of 35 cycles of denaturation at 947C for 30 s,
annealing at 537C for 40 s, and extension at 727C for 40 s.
The program for trnD-T consisted of 36 cycles of denaturation
 Table 1
Summary of the Data for Chamaecrista
Combined Combined Combined
(trnL-F 1 ITS) trnD-T trnL-F ETS ITS Morphology (cpDNA1 nDNA) (molecular 1 morphology)
No. of samples of Chamaecrista
ser. Coriaceae (species number) 33 (20) 40 (21) 46 (21) 44 (20) 35 (20) 36 (21) 36 (21) 36 (21)
No. of samples of outgroups 45 9 10 11 9 10 10 10
Total samples 78 49 56 55 44 46 46 46
Alignment length (bp) or no.
charactersa 1971 1121 1056 519 914 27 3470 3470
No. of indels coded
(simple coding) 346 31 68 85 131 ... 315 315
Total of characters (bp 1
indels coded) 2317 1152 1124 604 1045 27 3785 3785 1 27
No. of variable characters 474 77 130 156 238 1 637 637
No. of PI characters 692 51 51 242 307 26 585 612
CI, RI .59, .82 .89, .91 .91, .87 .70, .79 .73, .70 .47, .71 .74, .70 .72, .70
Tree length 2902 152 403/208 771 2524/1040 112 2145 2290
Nucleotide-substitution model MIXED TIM11I TVM1G TIM21G TIM31G/ NA MIXED MIXED
JC1G/TIM21G
Support for Coriaceae s.s. clade:
Boostrap (%) 79 94 90 !50 51 100b 99 99

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Posterior probability .92 .99 .81 .0 .0 NA 1 1
Decay index NA NA NA NA NA 5b NA NA
Note. Data presented are morphological data, sequence characteristics of the trnD-T, trnL-F, ETS, and ITS markers, and models applied in the Bayesian analyses for each molecular par-
tition. CI p consistency index; cpDNA p chloroplast DNA; ETS p external transcribed spacer; ITS p internal transcribed spacer; NA p not applicable; nDNA p nuclear DNA; PI p
parsimony-informative; RI p retention index; s.s. p sensu stricto.

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a
Number of characters is reported in the “Morphology” column.
b
Support resulted from analyses using only one species as the outgroup (Chamaecrista flexuosa).
 RANDO ET AL.—PHYLOGENY OF CHAMAECRISTA SER. CORIACEAE
947C for 50 s, annealing at 547C for 50 s, and extension at 727C
for 40 s.
Primers 17SE and 26SE were used for ITS amplification and
ITS92 and ITS4 primers for sequencing (Desfeaux et al. 1996);
the primers ETS IGS-760 (Baldwin and Markos 1998) and ETS
dioc3 (Queiroz et al. 2015) were used for amplification and se-
quencing of ETSs. Amplifications of both nuclear regions were
performed with a 30-mL PCR mix containing 3.0 mL of sup-
plied buffer, 1.5 mL of MgCl2 (50 mM), 0.6 mL of dNTPs (deoxy-
nucleotide triphosphates; 10 mM, GE Healthcare Life Sci-
ences), 0.3 mL of each primer (15mM), 0.6 mL of bovine serum
albumin (Invitrogen), 6.0 of mL betaine, 1.0 mL DMSO (1%,
Merck), 0.15 mL Taq DNA polymerase (5U/mL, Phoneutria),
15.85 mL H2O (q.s.p.), and 1.0 mL of DNA (20–40 ng). The pro-
gram for ITSs consisted of 28 cycles of denaturation 947C for 45 s,
annealing at 527C for 60 s, and extension at 727C for 180 s. The
program for ETSs consisted of 35 cycles of denaturation 947C
for 60 s, annealing at 527C for 60 s, and extension at 727C for
120 s.
PCR products were purified by enzymatic reactions with Exo-
nuclease I and shrimp alkaline phosphatase (Affymetrix, USB
Products, Cleveland) or with polyethylene glycol 20%. The sam-
ples were sequenced in both directions with a Spectrumedix SCI
SCE9624 automated sequencer at the Universidade Estadual
de Feira de Santana. In some cases, the PCR products were se-
quenced by Macrogen (Seoul).
Alignment and Phylogenetic Analyses
Forward and reverse sequencing reads were assembled into
contigs and edited with Geneious, version 6.1.7 (Biomatters
2013). Sequences of all loci were aligned with the MUSCLE
algorithm (Edgar 2004a, 2004b), with manual edition to cor-
rect obvious alignment errors and to remove sections with du-
bious alignments. The gaps in the matrices were coded with
the simple coding option (Simmons and Ochoterena 2000) in
SeqState, version 1.4.1. (Müller 2005). Individual and com-
bined analyses were performed for each molecular marker
and for the morphological data set. Ten different data matrices
were prepared in the following combinations: broad sampling:
(1) ITS, (2) trnL-F, and (3) ITS 1 trnL-F; narrow sampling:
(4) ETS, (5) ITS, (6) trnD-T, (7) trnL-F, (8) morphology, (9) all
molecular markers, and (10) all molecular markers and mor-
phology.
Maximum parsimony (MP) analyses were performed with
PAUP*, version 4.0b10 (Swofford 2003). A heuristic search
was conducted with 1000 random taxon-addition replicates
and the tree-bisection-reconnection (TBR) algorithm, saving
15 trees per replicate to prevent extensive swapping on islands
with many trees. The resulting trees were then used as starting
trees for a second-round search using TBR branch swapping
with an upper limit of 10,000 trees. Nonparametric bootstrap
support (BS) was estimated with 10,000 pseudoreplicates and
the same parameters used in our MP analyses (Felsenstein
1985). Decay indices were calculated only for the morpholog-
ical data set, and branch-and-bound search was performed in
PAUP*, version 4.0b10 (Swofford 2003).
Bayesian analyses (BAs) were performed with MrBayes 3.1
(Ronquist and Huelsenbeck 2003). Nucleotide-substitution mod-
els were selected, on the basis of the Akaike information cri-
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000
terion (AIC) values, with JModeltest 2.1 (Guindon and Gascuel
2003; Darriba et al. 2012). The substitution models selected for
each molecular marker are listed in table 1. Indels and morpho-
logical characters were coded as the standard characters “var-
iable” and “all,” respectively. The data sets were analyzed in
two independent runs, each with four chains. The individual
analyses used 5,000,000 generations, and the combined analy-
ses used 10,000,000 generations. Convergence was assessed
with Tracer, version 1.6 (Drummond and Rambaut 2007).
The sampled trees placed in the burn-in stage were excluded
(1000 for each marker and 10,000 for combined analyses),
and the remaining trees were assumed to be representative of
the posterior probability (PP) distribution.
Morphological Evolution
Morphological-character evolution was studied on the basis
of the majority-rule consensus tree derived from the BA of all
combined data sets. Selected accessions were trimmed so that
only a single terminal represented each monophyletic species. We
examined ancestral-state reconstructions for the same 27 charac-
ters that were used in the phylogenetic reconstruction (apps. B, C).
Reconstructions were performed in Mesquite 2.74 (Maddison
and Maddison 2010) with parsimony and maximum likelihood
criteria. The former used unordered character states and the lat-
ter a single-rate transition model (Mk1).
Bayesian Divergence-Age Estimation
Bayesian divergence-time estimation was performed with
BEAST, version 1.8.0 (Drummond et al. 2012), on the broad-
sampling data set. We used this data set because it covers
representatives of all major lineages of Chamaecrista and other
Cassiinae outgroups, thus allowing us to time-calibrate the node
of the most recent common ancestor (MRCA) of (Cassia 1
Senna 1 Chamaecrista) at 53 5 1.567 Ma, following previous
age estimations (Bruneau et al. 2008). A second calibration point
was based on a fossil fruit from the southeastern United States
and Mexico, described as Senna (Herendeen et al. 1992), also
used by Bruneau et al. (2008) and assigned to the stem node of
Senna species included in the analysis (the MRCA of Senna alata
and Senna gardneri), using a lognormal prior with a mean value
of 45.0 Ma and standard deviation of 0.005. The analyses
were performed with a tree prior obtained before a phyloge-
netic BA was performed. The data set included four partitions,
the plastid trnL-F spacer and the nuclear ITS split into three
regions (ITS 1, ITS 2, and the intervening 5.8S). Model param-
eters and molecular clocks were unlinked across partitions.
Nucleotide-substitution models (table 1) were selected on the
basis of AIC values with JModeltest 2.1 (Guindon and Gascuel
2003; Darriba et al. 2012). The analysis was performed with an
uncorrelated-rates, relaxed-molecular-clock model. Indepen-
dent runs were performed with 50,000,000 generations each.
The convergence of individual runs was assessed with Tracer,
version 1.6 (Drummond and Rambaut 2007). All analyses were
combined in LogCombiner, version 1.8.0 (Drummond et al.
2012), and the burn-in was established for the first 10% of trees.
All trees were summarized in a maximum clade credibility tree
with TreeAnnotator, version 1.8.0 (Drummond et al. 2012).
000 INTERNATIONAL JOURNAL OF PLANT SCIENCES
Results
All data sets resolved Chamaecrista ser. Coriaceae as poly-
phyletic in both MP analysis and BA. Despite this, the major-
ity of the species (18 species) formed a clade, named here the
Coriaceae clade. In all of the analyses, C. caribaea, C. rorai-
mae, and C. venulosa, included in Chamaecrista ser. Coriaceae
by Irwin and Barneby (1982), were always excluded from the
set of 18 species that includes C. choriophylla (Vogel) H.S.Irwin
& Barneby (the type of the series). The plastid data sets gave
stronger support for the monophyly of the Coriaceae clade than
the nuclear data sets, although the nuclear regions provided
greater resolution within the clade. The tree obtained from
the morphological data had no resolution, but combining all
of the data sets (molecular 1 morphological) increased sup-
port and resolution within the Coriaceae clade. We present be-
low the results obtained from analyses of plastid regions, nu-
clear regions, and the combined data. In addition, we present
the results of the reconstruction of the morphological evolu-
tion and of the Bayesian divergence-age estimation. Details
of DNA regions and morphological data sets, as well as the
phylogenetic analyses and support, are provided in table 1.
Broad-Sampling Data Set
There was no apparent incongruence between two regions
(ITS and trnL-F), but the ITS region gave greater resolution.
MP analysis and BA for the combined nuclear ITS 1 plastid
trnL-F data set resulted in the same topologies. The three spe-
cies that emerged outside the Coriaceae clade appeared scat-
tered in the large clade B (fig. 2), which includes species from
sections Chamaecrista and Xerocalyx. The Coriaceae clade
was recovered with moderate support (BS p 79, PP p 0.92;
see clade D in fig. 2); it emerged as a sister clade to C. swainsoni
(ser. Flexuosae).
Narrow-Sampling Data Set
All of the molecular markers analyzed individually, or com-
bined in several ways, with MP and Bayesian inference also re-
solved Chamaecrista ser. Coriaceae as polyphyletic, with no
incongruence among the regions. The Coriaceae clade was re-
covered with moderate to high support in all analyses. Both
plastid markers presented deletions exclusive to the Coriaceae
clade. There was a deletion of eight base pairs (GAGGGATA)
in the trnD-T region in the aligned positions 883–891, and there
was a large deletion of ca. 260 bp (positions 611–1017) in the
trnL-F region that could be recognized in the electrophoresis of
the PCR products. Both ETS and ITS data concurred in indicat-
ing C. aristata as the first-diverging species and in recognizing
a subordinated clade E comprising species with bifoliolate
leaves (fig. 3). In addition, ETS data allowed the recognition of
the moderately supported clade F, comprising mostly species
with multifoliate leaves.
The Coriaceae clade (clade D) was highly supported both in
MP analysis (BS p 99) and BA (PP p 1; fig. 3). This concat-
enated data set provided support (BS p 100, PP p 1) for
Chamaecrista ser. Flexuosae as a sister clade to the Coriaceae
clade. The subordinated clades E and F were recovered in BA
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of this data set, as were two additional clades within clade F,
although both with low support.
Morphological Evolution and Bayesian
Divergence-Age Estimation
Character-evolution reconstructions under MP and maximum
likelihood criteria tracked the same pattern, and we opted to pre-
sent the parsimony reconstructions in our results. Two of the 27
selected characters were indicated as putative synapomorphies
of the Coriaceae clade: the presence of a woody rootstock (char-
acter 2) and one stamen displaced toward the gynoecium and op-
posite to the other stamens (character 26; see fig. 4). Transitions
in six characters were indicated as putative synapomorphies
of subordinated clades within the Coriaceae clade: decumbent
shrubs (character 1), forked branches in the median region or
close to the apex (6), stipules perpendicular to the branch (12)
in a subclade of clade H, exclusively bifoliolate leaves (17), pal-
mate leaflet venation with more than four main veins and no
apical anastomosis (18) in clade H, and a terminal frondose-
bracteate raceme (22) in clade E (fig. 4).
In the Bayesian divergence-age analysis, estimated parameters
had adequate effective sample size (1200) after four runs. The
mean age estimate for the MRCA of the Coriaceae clade was
4.9 Ma (95% credibility interval: 8.06–2.76, highest posterior
density). From this point, the major diversification within the
clade was estimated around 1.3–3.4 Ma (fig. 2).
Discussion
Phylogenetic Relationships of Chamaecrista
sect. Chamaecrista ser. Coriaceae
All molecular markers (trnD-T, trnL-F, ETS, ITS), when con-
sidered together with morphological data, provided strong evi-
dence that Chamaecrista sect. Chamaecrista ser. Coriaceae is
monophyletic when recircumscribed to exclude three species
ascribed to it by Irwin and Barneby (1982): C. caribaea, C. ro-
raimae, and C. venulosa. Previous evidence for the monophyly
of the series (Conceição et al. 2009) was based on sparse sam-
pling that included only four species of the series now confirmed
to be part of the Coriaceae clade. Our analyses indicated that
the Coriaceae clade is embedded in a clade that unites the sec-
tions Chamaecrista, Caliciopsis, and Xerocalyx previously
found by Conceição et al. (2009). Analyses of the concatenated
data also yielded results similar to those of Conceição et al.
(2009) in supporting Chamaecrista ser. Flexuosae as sister to
the Coriaceae clade, although only two of the five species of
Chamaecrista ser. Flexuosae have been surveyed.
Several deletions observed in cpDNA were found exclusively
in species of the Coriaceae clade and are likely molecular syn-
apomorphies of this clade. In addition, at least two putative mor-
phological synapomorphies were found for the Coriaceae clade.
The presence of a woody rootstock is recurrent in Chamaecrista,
mainly in species of Chamaecrista sect. Absus that occur in Cer-
rado and Campo Rupestre vegetation. A special underground
system like this probably evolved independently in distinct line-
ages of the genus during their specialization in fire-prone habitats
such as the Cerrado and Campo Rupestre (Irwin and Barneby
1978, 1982). However, within Chamaecrista sect. Chamaecrista
 Fig. 2 Maximum clade credibility tree for Chamaecrista from Bayesian analysis in BEAST. The highest posterior density age estimates for
main nodes are presented in bars, with a timescale on the X-axis. Legend on left depicts the posterior probability obtained for each node in the
Bayesian analysis. Numbers below some branches indicate bootstrap support (left, in percentage) and posterior probability (right, in decimal
form) for the clades that also emerged in phylogenetic analyses of broad sampling (ITS 1 trnL-F). Clades highlighted with boxed capital letters
are discussed in the text. The type species of Chamaecrista ser. Coriaceae (Chamaecrista choriophylla) is indicated in bold type. Infrageneric
groups as recognized by Irwin and Barneby (1982) are indicated. BA p Bahia; MG p Minas Gerais.

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 Fig. 3 Narrow sampling of Chamaecrista: majority-rule consensus tree derived from Bayesian analysis of the combined data set (external
transcribed spacer, ITS, trnd-T, trnl-F, and morphology). Numbers below the branches indicate bootstrap support (left, in percentage) values for
the clades that also emerged in the maximum parsimony analysis, along with posterior probability (right, in decimal form). Asterisks indicate
species traditionally recognized in Chamaecrista ser. Coriaceae. Clades marked by boxed capital letters are discussed in the text. BA p Bahia;
ES p Espírito Santo; MG p Minas Gerais; RR p Roraima.

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 RANDO ET AL.—PHYLOGENY OF CHAMAECRISTA SER. CORIACEAE 000

Fig. 4 Ancestral-state reconstruction of eight selected morphological characters in Chamaecrista. The vertical bars represent character
transitions according to parsimony reconstruction. Characters 2 and 26 represent putative synapomorphies of the Coriaceae clade. The others
six characters represent putative synapomorphies of subordinate clades within the Coriaceae clade. For more details concerning the characters
and the selected character states, see appendixes B and C, available online.

(composed of ca. 52 Brazilian species), the Coriaceae clade is the
only group that consistently produces a woody rootstock. These
woody rootstocks (Radford et al. 1976) are complex under-
ground structures whose morphological nature, anatomy, and
ontogeny are still poorly studied (Appezzato-da-Glória 2003),
and so we avoid here the application of the term “xylopodium”
as used by Irwin and Barneby (1982). Moreover, overall mor-
phological features described as woody rootstocks may not be
homologous in Chamaecrista; additional anatomical and devel-
opmental studies will be necessary to study the homology of
these woody underground root systems.
The unique floral structure, with one stamen displaced to the
same side as the gynoecium and away from the other stamens
(see fig. 5), is another possible synapomorphy of the Coria-
ceae clade. This floral pattern has not been observed in the re-
maining Brazilian species of Chamaecrista, and only the en-
demic Mexican C. chamaecristoides (Collad.) Greene shows a
similar configuration, although in a consistently heteromorphic
androecium (Arceo-Gomes et al. 2012).
The former circumscription of Chamaecrista ser. Coriaceae
(Irwin and Barneby 1982) included species with shrubby habits,
a woody rootstock, thick-textured leaflets, axillary peduncles,

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 Fig. 5 Flower morphology diversity in Chamaecrista. A, C. lineata: largest petal not curved, heteromorphic androecium (C. caribaea has similar
morphology). B, C. roraimae: no evident differentiated petal (five petals equal), homomorphic androecium. C, D, C. venulosa (C) and C. flexuosa
(D): slightly curved petal, homomorphic androecium. E, F, C. distichoclada (E) and C. papillata (F): strongly curved petal, homomorphic androe-
cium, and one stamen displaced to opposite side from the other stamens. G, H, Underground structure: G, C. caribaea var. lucayana; H, C. rorai-
mae. I, C. choriophylla. J, C. olesiphylla. K, C. rotundata var. grandistipula. Scale bars p 1 cm.

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 RANDO ET AL.—PHYLOGENY OF CHAMAECRISTA SER. CORIACEAE
and extrafloral nectaries. Fieldwork and herbarium studies have
revealed that even though characters such as a shrubby habit,
axillary peduncles, and extrafloral nectaries occur in all species
of the series, they are also present in species of other sections of
Chamaecrista. We noted, on the other hand, that leaflet tex-
ture is highly variable among the species of this series, and the
presence of a woody rootstock cannot be definitely ascribed
to any species without detailed field observations or access to
complete herbarium material. However, on the basis of field
observations of the root systems of the three species ascribed
to Chamaecrista ser. Coriaceae that were identified here as not
being related to the Coriaceae clade, we can confirm that they
do not have woody rootstocks (fig. 5), contrary to the reports
of Irwin and Barneby (1982).
In addition to the absence of a woody rootstock, the three
species traditionally placed in Chamaecrista ser. Coriaceae
that did not appear nested in the Coriaceae clade in our study
are easily distinguished from other Coriaceae species by a num-
ber of other characteristics. Chamaecrista caribaea was re-
solved as part of Chamaecrista ser. Chamaecrista, a group tra-
ditionally recognized by having peduncles adnate to the stems
(and thus appearing supra-axillary to the leaf axil). However,
we did not observe this trait in C. caribaea or in the sister and
sympatric C. lineata, suggesting that peduncle adnation to the
stem could be phylogenetically labile. Both C. caribaea and
C. lineata share a particular floral morphology, with a hetero-
morphic androecium and a labellum-like petal (fig. 5), and
also have penninerved leaflet venation, very similar to that of
C. pascuorum, C. repens, and C. nictitans (which appeared as
closely related in all of our analyses). Chamaecrista roraimae
displays a different floral morphology, with a homomorphic an-
droecium and no differentiated internal petal, and emerges as
closely related to C. caliciopsis (which traditionally constituted
the monospecific Chamaecrista sect. Caliciopsis). Chamaecrista
venulosa occurs sympatrically with species of the Coriaceae
clade in the Espinhaço Range. It has a homomorphic androe-
cium and a differentiated petal but does not exhibit a displaced
stamen (fig. 5). Chamaecrista venulosa is resolved as closely re-
lated to species of Chamaecrista ser. Prostratae (fig. 2).
Two major groups appeared within the Coriaceae clade, al-
though with low support: the “bifoliolate clade,” composed
of species with only one pair of leaflets, and the “multifolio-
late clade,” composed of species with two or more pairs of
leaflets (figs. 3, 5). This reduction in the number of leaflets
seems a putative synapomorphy for the bifoliolate clade. Other
morphological characteristics observed in this bifoliolate clade
include a decumbent habit (with a reversion to an erect habit
in C. lagotois) and a terminal frondose-bracteate inflorescence
characterized by several axillary flowers placed on the distal
portions of branches bearing reduced leaves. Two other groups
are found within the multifoliolate clade: clade G, grouping
plants conserving symplesiomorphic characters such as erect
stipules with truncate bases and palmate veined leaflets (in this
case, 3 or 4 veins with apical anastomosis), and clade H, group-
ing plants with synapomorphic characters such as stipules per-
pendicular to the axis and leaflets with a particular palmate
venation pattern. The relationships of C. aristata and C. anceps
to the remaining species of the Coriaceae clade are unclear. The
former could be the sister species to the rest of the clade, while
the position of C. anceps is uncertain.
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000
The position of the stipules (erect and adnate to the branches vs.
perpendicular to the branch) seems to be correlated with stipule
size. Large stipules are adnate to the branches only on the distal
portions of the branch, and subsequently they gradually change
to a completely perpendicular position (fig. 5) or are ultimately
caducous. In the perpendicular position, larger stipules cover-
ing apical portions of branches expose the extrafloral nectary
on petioles, and these are often associated with ants. These large
stipules are commonly observed in Cerrado species of several
plant families and are probably related to protection against
fire (Simon and Pennington 2012) and/or herbivory. Smaller
stipules that do not change their positions through time poten-
tially provide less protection against fire, while the nectaries
on young leaves are completely exposed throughout the leaf de-
velopment and thus more available to ants.
Diversification of the Coriaceae Clade
in Campo Rupestre Vegetation
The divergence-age analysis estimated a recent in situ diversi-
fication for the Coriaceae clade of 3.4–1.3 Ma in the Campo
Rupestre vegetation of the Espinhaço Range. As reported here
for the Coriaceae clade, phylogenetic studies in other Campo
Rupestre lineages have detected similar patterns, in which most
of the diversity arose by in situ diversification from a limited
number of radiations, notably in the Monticola clade of Cal-
liandra (Leguminosae; Souza et al. 2013), Paepalanthus (Erio-
claulaceae; Trovó et al. 2013), Minaria (Apocynaceae; Ribeiro
et al. 2014), and Hoffmannseggella (Orchidaceae; Antonelli
et al. 2010). In all cited groups, age estimation showed that di-
versification took place within the past 5 Myr.
Because of its fire-prone nature and open vegetation landscape,
the Campo Rupestre has usually been considered part of the
Cerrado, the largest area of savanna vegetation in South Amer-
ica (e.g., Simon et al. 2009; Trovó et al. 2013). The flora of the
Cerrado was assembled from numerous independent plant line-
ages from different biomes, mostly during the past 4–5 Myr (Si-
mon et al. 2009), in line with age estimates for Campo Rupestre
lineages, including the Coriaceae clade. However, in contrast to
Cerrado lineages, Campo Rupestre groups show higher levels of
endemism, tend to present phylogenies that are geographically
structured at finer scales (i.e., across several separate mountains;
Hughes et al. 2013; Trovó et al. 2013), and have higher species
turnover, inferred from both higher speciation and higher extinc-
tion rates (Koenen et al. 2013). This indicates that Cerrado and
Campo Rupestre floras may have been subjected to distinct bio-
geographical processes despite their overall landscape similarity
and interdigitated geography. The dissected nature and small
areas where the Campo Rupestre occur could act as biogeo-
graphical islands, as demonstrated by population-level studies
showing fine-scale genetic disjunctions caused by microhabitat
heterogeneity (e.g., Borba et al. 2001; Jesus et al. 2001; Lambert
et al. 2006; Pereira et al. 2007; Ribeiro et al. 2008).
The presence of a woody underground system was demon-
strated as a morphological synapomorphy of the Coriaceae
clade. Woody rootstocks (including xylopodia) are regarded
as an adaptation to regular fire regimes, allowing the plant to
resprout after burning of the aerial parts (Gottsberger and
Silberbauer-Gottsberger 2006; Simon and Pennington 2012).
Thus, it is possible that this structure could represent a key in-
000 INTERNATIONAL JOURNAL OF PLANT SCIENCES

novation in the Coriaceae clade allowing its diversification in Acknowledgments

the Campo Rupestre environment.
In conclusion, both molecular and morphological data indi-
cated that a new, slightly narrower definition of Chamaecrista
ser. Coriaceae is required for it to be recognized as a monophyletic
group, excluding C. caribaea, C. roraimae, and C. venulosa. In
this new circumscription, the series Coriaceae comprises 19
shrubby species with woody rootstocks and an androecium with
one stamen displaced to the same side as the gynoecium (and
away from the remaining stamens); all species are restricted to
highland areas of central-eastern Brazil, mainly to Campo Ru-
pestre vegetation. Even though the phylogeny presented here
provides the evidence for a new circumscription of the group,
further work is necessary to improve resolution of species re-
lationships and to achieve higher support for the groups within
the Coriaceae clade.
We would like to thank the herbaria cited for granting ac-
cess to their collections and Ana Maria Goulart de Azevedo
Tozzi, Anselmo Nogueira, Luiz Martins Fonseca, Marcelo F. Si-
mon, Vidal Mansano, and Vinícius Castro Souza for the helpful
comments they offered on the manuscript. We also would like
to thank the anonymous reviewers, Gwilym Lewis, and Patrick
Herendeen for the revisions offered. This article is part of the
PhD thesis of J. G. Rando at the Universidade de São Paulo. Fi-
nancial support was provided to J. G. Rando by FAPESP (process
2009/16434–4 and 2013/09374–0), and to L. P. de Queiroz and
J. R. Pirani by CNPq. The molecular steps of this research were
also funded by SISBIOTA (CNPq process 563084/2010–3 and
FAPESB PES0053/2011), CAPES (AUXPE-PROEX 640/2011),
and PRONEX (CNPQ/FAPESB grant PNX0014/2009).

Appendix A

Voucher Information and GenBank Accession Numbers of the Sequences Included in This Study

Taxon, (accession number for duplicates), origin; voucher, herbarium, and GenBank accession numbers for ITS, ETS, trnD-T,
and trnL-F. Sequences not generated in this study are marked with an asterisk.
Cassia grandis L.f., Brazil; Queiroz 2878, HUEFS, —, FJ009820,* —, FJ009875.* Cassia javanica L., Brazil; Queiroz 11039,
HUEFS, —, FJ009821,* —, FJ009876.* Chamaecrista absus (L.) H.S.Irwin & Barneby var. absus, Brazil; Conceição 1056,
HUEFS, —, FJ009832,* —, FJ009886.* Chamaecrista amorimii Barneby, Brazil; Conceição 795, HUEFS, KP967071,
FJ009823,* KP966952, FJ009878.* Chamaecrista anamariae Conc., L.P. Queiroz & G.P. Lewis, Brazil; Conceição 787,
HUEFS, —, FJ009826,* —, FJ009881.* Chamaecrista anceps (Benth.) H.S.Irwin & Barneby, Brazil; Cota 410, DIAM, —,
—, KP966953, KP966898. Chamaecrista aristata (Benth.) H.S.Irwin & Barneby, Brazil; Rando 976, SPF, KP967070,
KP967072, KP966954, KP966899. Chamaecrista belemii (H.S.Irwin & Barneby) H.S.Irwin & Barneby var. belemii, Brazil;
Queiroz 9151, HUEFS, —, DQ787389,* —, FJ009880.* Chamaecrista blanchetii (Benth.) Conc., L.P. Queiroz & G.P. Lewis,
Brazil; Andrade 607, HUEFS, —, FJ009846,* —, FJ009900.* Chamaecrista botryoides Conc., L.P. Queiroz & G.P. Lewis,
Brazil; Conceição 541, HUEFS, —, FJ009836,* —, FJ009890.* Chamaecrista brachystachya (Benth.) Conc., L.P. Queiroz &
G.P. Lewis, Brazil; Conceição 713, HUEFS, KP967069, FJ009847,* KP966955, FJ009901.* Chamaecrista burchellii (Benth.)
H.S.Irwin & Barneby, Brazil; Rando 1092, HUEFS, SPF, KP967068, KP967073, KP966956, KP966900. Chamaecrista
calycioides (DC. ex Collad.) Greene, Brazil; Queiroz 11, HUEFS, —, FJ009863,* —, FJ009917.* Chamaecrista campestris
H.S.Irwin & Barneby, Brazil; Queiroz 10440, HUEFS, —, FJ009829,* —, FJ009883.* Chamaecrista cardiostegia H.S.Irwin
& Barneby, Brazil; Rando 1125, SPF, KP967067, KP967074, KP966957, KP966901. Chamaecrista caribaea var. lucayana
(Britton) H.S.Irwin & Barneby, Bahamas; Rando 963, NY, SPF, KP967066, KP967075, KP966958, KP966902. Chamaecrista
cathartica (Mart.) H.S.Irwin & Barneby, Brazil; Conceição 789, HUEFS, KP967065, FJ009841,* —, FJ009895.* Chamaecrista
choriophylla (Vogel) H.S.Irwin & Barneby, Brazil; Rando 1034, HUEFS, KP967063, KP967076, KP966959, KP966904. C.
choriophylla (Vogel) H.S.Irwin & Barneby, (1), Brazil; Rando 907, SPF, KP967064, —, —, KP966903. Chamaecrista
cinerascens (Vogel) H.S.Irwin & Barneby, Brazil; Rando 661, SPF, KP967062, KP967077, KP966960, KP966905. Chamaecrista
confertiformis (H.S.Irwin & Barneby) Conc., L.P. Queiroz & G.P. Lewis, Brazil; Costa 132, HUEFS, —, FJ009848,* —,
FJ009902.* Chamaecrista coriacea (Bong. ex Benth.) H.S.Irwin & Barneby, Brazil; Conceição 869, HUEFS, —, FJ009843,*
—, FJ009897.* Chamaecrista cytisoides (DC. ex Collad.) H.S.Irwin & Barneby, Brazil; Conceição 870, HUEFS, —,
FJ009844,* —, FJ009898.* Chamaecrista dalbergiifolia (Benth.) H.S.Irwin & Barneby, Brazil; Queiroz 10318, HUEFS, —,
FJ009837,* —, FJ009891.* Chamaecrista decora (H.S.Irwin & Barneby) Conc., L.P. Queiroz & G.P. Lewis, Brazil; Conceição
810, HUEFS, —, FJ009849,* —, FJ009903.* Chamaecrista depauperata Conc., L.P. Queiroz & G.P. Lewis, Brazil; Conceição
863, HUEFS, —, FJ009850,* —, FJ009904.* Chamaecrista desvauxii var. desvauxii (Collad.) Killip, Brazil; Queiroz 10453,
HUEFS, —, FJ009864,* —, FJ009918.* C. desvauxii var. langsdorffii (Kunth ex Vogel) Britton ex Pittier, Brazil; Conceição
674, HUEFS, —, FJ009866,* —, FJ009920.* C. desvauxii var. latistipula (Benth.) G.P.Lewis, Brazil; Conceição 912, HUEFS,
KP967061, FJ009867,* KP967008, FJ009921.* C. desvauxii var. mollissima (Benth.) H.S.Irwin & Barneby, Brazil; Santos
356, HUEFS, —, FJ009865,* —, FJ009919.* Chamaecrista diphylla (L.) Greene, Brazil; Queiroz 10269, HUEFS, —,
FJ009868,* —, FJ009922.* Chamaecrista distichoclada (Benth.) H.S.Irwin & Barneby, Brazil; Rando 1230, HUEFS, SPF,
KP967060, KP967078, KP966961, KP966906. Chamaecrista flexuosa (L.) Greene, Brazil; Barbosa 1145, HUEFS, KP967059,
—, KP966962, —. C. flexuosa (L.) Greene, Brazil; Giulietti 2344, HUEFS, —, FJ009858,* —, FJ009912.* Chamaecrista
glaucofilix (H.S.Irwin & Barneby) H.S.Irwin & Barneby, Brazil; Conceição 861, HUEFS, —, FJ009834,* —, FJ009888.*
Chamaecrista hispidula (Vahl) H.S.Irwin & Barneby, Brazil; Conceição 914, HUEFS, —, FJ009833,* —, FJ009887.*

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 RANDO ET AL.—PHYLOGENY OF CHAMAECRISTA SER. CORIACEAE 000

Chamaecrista jacobinea (Benth.) H.S.Irwin & Barneby, Brazil; Andrade 610, HUEFS, —, FJ009827,* —, FJ009882.*
Chamaecrista lagotois H.S.Irwin & Barneby, Brazil; Rando 1029, HUEFS, KP967058, KP967079, KP966963, KP966907. C.
lagotois H.S.Irwin & Barneby, (1), Brazil; Rando 1133, SPF, KP967057, KP967080, —, KP966908. Chamaecrista latifolia
(Benth.) Rando, Brazil; Rando 1024, HUEFS, KP967056, KP967081, KP966964, KP966909. C. latifolia (Benth.) Rando, (1),
Brazil; Rando 1127, SPF, KP967055, KP967082, KP966965, KP966910. Chamaecrista lineata (Swartz.) Greene var. lineata,
Bahamas; Rando 958, SPF, KP967085, —, KP966967, KP966912. C. lineata (Swartz.) Greene var. lineata, (1), Bahamas; Rando
954, SPF, KP967084, —, —, —. Chamaecrista mucronata (Spreng.) H.S.Irwin & Barneby, (MG), Brazil; Rando 879, SPF,
KP967050, KP967086, KP966968, KP966913. C. mucronata (Spreng.) H.S.Irwin & Barneby, (ES), Brazil; Siqueira 825,
HUEFS, KP967049, —, KP966969, KP966914. Chamaecrista nictitans subsp. brachypoda (Benth.) H.S.Irwin & Barneby, Brazil;
Queiroz 10335, HUEFS, —, FJ009855,* —, FJ009909.* C. nictitans var. disadena (Steud.) H.S.Irwin & Barneby, Brazil;
Conceição 790, HUEFS, —, FJ009852,* —, FJ009906.* C. nictitans subsp. patellaria var. ramosa (Vogel) H.S.Irwin & Barneby,
Brazil; Queiroz 10406, HUEFS, —, FJ009853,* —, FJ009907.* Chamaecrista olesiphylla (Vogel) H.S.Irwin & Barneby, Brazil;
Rando 1147, SPF, KP967047, KP967089, KP966972, KP966916. C. olesiphylla (Vogel) H.S.Irwin & Barneby, (1), Brazil;
Rando 632, SPF, KP967046, KP967088, KP966971, KP966915. C. olesiphylla (Vogel) H.S.Irwin & Barneby, (2), Brazil; van
den Berg 1350, HUEFS, KP967045, KP967091, KP966974, KP966918. Chamaecrista onusta H.S.Irwin & Barneby, Brazil;
Conceição 800, HUEFS, —, FJ009824,* —, FJ009879.* Chamaecrista papillata H.S.Irwin & Barneby, (MG), Brazil; Rando
1011, HUEFS, KP967043, KP967093, KP966976, KP966920. C. papillata H.S.Irwin & Barneby, (BA 1), Brazil; Borba
1922, HUEFS, KP967044, KP967095, KP966978, —. C. papillata H.S.Irwin & Barneby, (BA 2), Brazil; Rando 1109, HUEFS,
SPF, KP967042, KP967094, KP966977, KP966921. Chamaecrista pascuorum (Benth.) H.S.Irwin & Barneby, Brazil; Queiroz
9169, HUEFS, —, FJ009851,* KP966979, FJ009905.* Chamaecrista philippi (H.S.Irwin & Barneby) H.S.Irwin & Barneby,
Brazil; Giulietti 2245, HUEFS, —, FJ009838,* —, FJ009892.* Chamaecrista pilosa (L.) Greene, Brazil; Queiroz 10221, HUEFS,
—, FJ009856,* —, FJ009910.* Chamaecrista potentilla (Benth.) H.S.Irwin & Barneby, Brazil; Rando 814, SPF, KP967039, —,
—, KP966922. C. potentilla (Benth.) H.S.Irwin & Barneby, Brazil; Queiroz 7606, HUEFS, KP967040, KP967097, —,
FJ009913,* C. potentilla (Benth.) H.S.Irwin & Barneby, (1), Brazil; Rando 1139, SPF, KP967038, KP967096, KP966980,
KP966923. C. potentilla (Benth.) H.S.Irwin & Barneby, (2), Brazil; Rando 1245, SPF, KP967037, —, KP966981, KP966924.
Chamaecrista repens (Vogel) H.S.Irwin & Barneby, Brazil; Giulietti 2325, HUEFS, —, KP967098, KP966982, KP966925. C.
repens (Vogel) H.S.Irwin & Barneby, Brazil; Carvalho 48, HUEFS, KP967036, —, —, KP966926. Chamaecrista roraimae
(Benth.) Gleason, (RR), Brazil; Rando 1154, SPF, KP967033, KP967099, KP966983, KP966927. C. roraimae (Benth.) Gleason,
(BA), Brazil; Harley 55036, HUEFS, KP967034, —, KP966984, KP966928. C. roraimae (Benth.) Gleason, (BA 1), Brazil; Harley
54793, HUEFS, KP967035, —, KP966985, —. Chamaecrista rossicorum (H.S.Irwin & Barneby) Rando, Brazil; Conceição 539,
HUEFS, KP967032, KP967102, KP966989, KP966932. C. rossicorum (H.S.Irwin & Barneby) Rando, (1), Brazil; Rando 1045,
HUEFS, KP967029, KP967101, KP966988, KP966931. C. rossicorum (H.S.Irwin & Barneby) Rando, (2), Brazil; Rando 1007,
HUEFS, —, KP967100, KP966987, KP966930. Chamaecrista rotundata var. grandistipula (Vogel) H.S.Irwin & Barneby, Brazil;
Rando 1240, SPF, KP967025, KP967106, KP966994, KP966937. C. rotundata var. grandistipula (Vogel) H.S.Irwin & Barneby,
(1), Brazil; Rando 994, HUEFS, KP967027, KP967105, KP966992, KP966935. C. rotundata var. grandistipula (Vogel)
H.S.Irwin & Barneby, (2), Brazil; Rando 925, K, SPF, KP967028, KP967104, KP966991, KP966934. C. rotundata (Vo-
gel) H.S.Irwin & Barneby var. rotundata, Brazil; Rando 1144, SPF, KP967022, KP967103, KP966990, KP966933. C. rotundata
var. interstes H.S.Irwin & Barneby, Brazil; Rando 1145, SPF, KP967024, KP967107, KP966995, KP966938. C. rotundata var.
interstes H.S.Irwin & Barneby, (1), Brazil; Rando 1236, SPF, KP967023, KP967108, KP966996, KP966939. C. rotundata
var. rotundata (Vogel) H.S.Irwin & Barneby, (2), Brazil; van den Berg 1333, HUEFS, KP967021, —, —, KP966941.
Chamaecrista rotundifolia var. grandiflora (Pers.) Greene, Brazil; Costa 128, HUEFS, —, FJ009857,* —, FJ009911.*
Chamaecrista rupestrium H.S.Irwin & Barneby, Brazil; Santos 390, HUEFS, —, FJ009835,* —, FJ009889.* Chamaecrista
setosa var. dentosa (Benth.) H.S.Irwin & Barneby, Brazil; Queiroz 10460, HUEFS, —, FJ009842,* KP966998, FJ009896.*
Chamaecrista simplifacta H.S.Irwin & Barneby, Brazil; Rando 802, SPF, KP967019, KP967109, KP966999, KP966942. C.
simplifacta H.S.Irwin & Barneby, (1), Brazil; Rando 1148, SPF, KP967018, KP967110, KP967000, KP966943. Chamaecrista
speciosa Conc., L.P.Queiroz & G.P.Lewis, Brazil; Conceição 546, HUEFS, —, FJ009839,* —, FJ009893.* Chamaecrista
supplex (Mart. ex Benth.) Britton & Rose, Brazil; Queiroz 10217, HUEFS, —, FJ009869,* —, FJ009923.* Chamaecrista
swainsoni (Benth.) H.S.Irwin & Barneby, Brazil; Conceição 822, HUEFS, KP967017, —, —, —. C. swainsoni (Benth.)
H.S.Irwin & Barneby, (1), Brazil; Queiroz 12314, HUEFS, KP967016, KP967111, KP967001, KP966944. Chamaecrista
tragacanthoides var. rasa H.S.Irwin & Barneby, Brazil; Rando 969, SPF, KP967015, KP967113, KP967003, KP966946. C.
tragacanthoides var. rasa H.S.Irwin & Barneby, (1), Brazil; Rando 1005, SPF, KP967014, KP967087, KP966970,
KP966947. C. tragacanthoides (Benth.) H.S.Irwin & Barneby var. tragacanthoides, Brazil; Pirani 6334, SPF, KP967013,
KP967114, KP967004, KP966948. Chamaecrista ulmea H.S.Irwin & Barneby, Brazil; Santos 650, SPF, KP967012,
KP967115, KP967005, KP966949. Chamaecrista unijuga (Benth.) Conc., L.P.Queiroz & G.P.Lewis, Brazil; Conceição 694,
HUEFS, —, FJ009845,* —, FJ009899.* Chamaecrista urophyllidia (H.S.Irwin & Barneby) H.S.Irwin & Barneby, Brazil; Harley
54656, HUEFS, —, FJ009840,* —, FJ009894.* Chamaecrista venulosa (Benth.) H.S.Irwin & Barneby, (MG 1), Brazil; Rando
1015, HUEFS, KP967011, KP967116, KP967006, KP966950. C. venulosa (Benth.) H.S.Irwin & Barneby, (MG 2), Brazil; Snak
671, HUEFS, KP967009, —, —, KP966951. C. venulosa (Benth.) H.S.Irwin & Barneby, (BA), Brazil; Rando 1256, HUEFS,
KP967010, —, KP967007, —. Senna alata (L.) Roxb., Brazil; HUEFS, —, HQ33041,* —, —. Senna gardneri (Benth.) H.S.Irwin
& Barneby, Brazil; Queiroz 7860, HUEFS, —, FJ009822,* —, FJ009877.*

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000 INTERNATIONAL JOURNAL OF PLANT SCIENCES
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