Tracing the impact of the Andean uplift on Neotropical plant evolution

Tracing the impact of the Andean uplift on Neotropical plant evolution
Tracing the impact of the Andean uplift on
Neotropical plant evolution
Alexandre Antonellia,1,2, Johan A. A. Nylanderb, Claes Perssona, and Isabel Sanmartı́nc,2
aDepartment of Plant and Environmental Sciences, University of Gothenburg, Box 461, 405 30 Gothenburg, Sweden; bDepartment of Botany, Stockholm
University, 106591 Stockholm, Sweden; and cDepartment of Biodiversity and Conservation, Real Jardı́n Botánico, Consejo Superior de Investigaciones
Cientificas, Plaza de Murillo 2, 28014 Madrid, Spain
Recent phylogenetic studies have revealed the major role played
by the uplift of the Andes in the extraordinary diversification of the
Neotropical flora. These studies, however, have typically considered the Andean uplift as a single, time-limited event fostering the
evolution of highland elements. This contrasts with geological
reconstructions indicating that the uplift occurred in discrete periods from west to east and that it affected different regions at
different times. We introduce an approach for integrating Andean
tectonics with biogeographic reconstructions of Neotropical
plants, using the coffee family (Rubiaceae) as a model group. The
distribution of this family spans highland and montane habitats as
well as tropical lowlands of Central and South America, thus
offering a unique opportunity to study the influence of the Andean
uplift on the entire Neotropical flora. Our results suggest that the
Rubiaceae originated in the Paleotropics and used the boreotropical connection to reach South America. The biogeographic patterns found corroborate the existence of a long-lasting dispersal
barrier between the Northern and Central Andes, the ‘‘Western
Andean Portal.’’ The uplift of the Eastern Cordillera ended this
barrier, allowing dispersal of boreotropical lineages to the South,
but gave rise to a huge wetland system (‘‘Lake Pebas’’) in western
Amazonia that prevented in situ speciation and floristic dispersal
between the Andes and Amazonia for at least 6 million years. Here,
we provide evidence of these events in plants.
biogeography 兩 Neotropical biodiversity 兩 Rubiaceae
T
he uplift of the tropical Andes in the Neogene had a
profound impact on the history of the South American
continent. It changed the course of the Amazon system from
flowing northwestwards to the modern system that flows to the
Atlantic side (1, 2) and affected the climate of the region by
forming the only barrier to atmospheric circulation in the
Southern Hemisphere (3). Recent phylogenetic studies have
shown that the Andean orogeny had also a major role in the
evolution of the Neotropical flora. The Neotropics hold the
highest plant species diversity in the world (4). This richness has
traditionally been explained in terms of environmental factors
(5), but lately, more integrative explanations have been advanced
that emphasize the role of historical and evolutionary factors in
the shaping of Neotropical diversity (6, 7). The ‘‘tropical conservatism hypothesis,’’ for example, argues that there are more
plant species in the Neotropics simply because more lineages
originated and diversified there, owing to the long-term climatic
stability of the region and the tendency of species to retain their
climatic niches over evolutionary time (7, 8). It is now also clear
that part of this richness has been gained by the migration of
lineages from other biogeographic regions (6). For instance,
pantropically distributed plant families such as Malpighiaceae,
Fabaceae, and Annonaceae (6, 9, 10, 11) originated at temperate
latitudes as part of the former ‘‘boreotropical flora’’ (12–14) and
subsequently entered the Neotropics via the mountain ranges of
Central America and the newly formed Northern Andes. One
point in common to these hypotheses is the key role that the
formation of the tropical Andes would have played in the
www.pnas.org兾cgi兾doi兾10.1073兾pnas.0811421106
historical diversification of the Neotropical flora (15). Recent
phylogenetic studies have shown that the Andean uplift acted
both as a dispersal route for boreotropical lineages (16, 17) and
as a driver in promoting rapid diversification, via allopatric
speciation and ecological displacement, in highland (16–19) and
montane (11) habitats.
Fewer studies, however, have documented the impact of the
Andean uplift on the lowland Amazonian flora. Clearly, the
uplift must have affected these taxa by forming a new biotic
barrier and profoundly changing the hydrology and climate of
the region (20). Furthermore, previous biogeographic studies on
Andean radiations have typically considered the Andean orogeny as a single, time-limited event, usually in connection with the
final (Miocene to Pleistocene) uplift of the Andes (11, 19). This
contrasts with geological reconstructions indicating that the
uplift took place in discrete periods, progressing from south to
north and from west to east and affecting different regions at
different times (2, 3, 21, 22). Episodic marine incursions, related
to global sea level rises during the extensional tectonic phases
that followed periods of major uplift, had a dramatic impact in
the drainage patterns of the region, as evidenced by paleogeographic and paleontological evidence (1, 2, 23–28). These marine
incursions have been discussed in relation to their role as a
pathway in the evolutionary transition from marine to freshwater
habitats of Neotropical fishes (24, 29), but they could also have
acted as barriers to dispersal or as vicariance events fragmenting
the ranges of terrestrial animals and plants. It seems surprising
that, despite increasingly detailed geological reconstructions (2,
24, 26–28), thus far no study has attempted to document the
effect of these events on the evolution of the Neotropical flora.
Generally, detailed reconstructions have been hampered by the
lack of resolution in many Andean species-rich clades (19).
Current biogeographic methods require well-resolved phylogenies, and uncertainty in phylogenetic relationships makes it
difficult to reconstruct the specific sequence of geological vicariance and speciation events.
Here, we use an integration of phylogenetic, biogeographic,
and molecular dating methods to reconstruct the evolutionary
history of tribes Cinchoneae and Isertieae, which together form
one of the major clades of Neotropical Rubiaceae. The distriAuthor contributions: A.A. and I.S. designed research; A.A. and C.P. performed research;
J.A.A.N. and I.S. contributed new reagents/analytic tools; A.A., J.A.A.N., and I.S. analyzed
data; and A.A. and I.S. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission. B.H.T. is a guest editor invited by the Editorial
Board.
Freely available online through the PNAS open access option.
Data deposition: The sequences reported in this paper has been deposited in the GenBank
database (accession nos. DQ448595–DQ448612).
1Present address: Institute of Systematic Botany, University of Zurich, Zollikerstrasse 107, CH
8008, Zurich, Switzerland.
2To
whom correspondence may be addressed. E-mail: [email protected] or
[email protected]
This article contains supporting information online at www.pnas.org/cgi/content/full/
0811421106/DCSupplemental.
PNAS Early Edition 兩 1 of 6
EVOLUTION
Edited by Bruce H. Tiffney, University of California, and accepted by the Editorial Board April 13, 2009 (received for review November 11, 2008)
bution of this clade spans highland and montane habitats (the
Andes, the Guiana, and the Brazilian Shields), as well as lowland
tropical forests (the Amazonia and Chocó). It thus offers a
unique opportunity to disentangle the evolutionary processes
underlying botanical evolution in the region. Our results reveal
an extraordinary level of congruence between the evolution of
the Neotropical Rubiaceae and the progressive west-to-east
Andean uplift, which brought about a series of marine incursions
and lacustrine systems that blocked the dispersal of plants and
shaped the distribution of the modern flora.
Study Group
The coffee family (Rubiaceae) is the fourth largest family of
flowering plants, with some 13,100 species in 611 genera and 3
subfamilies (30, 31). Although cosmopolitan in distribution, its
highest diversity is distinctly confined to the tropics. Subfamily
Rubioideae is pantropically distributed and comprises some
highly diverse groups in the Neotropics (e.g., Palicoureeae and
Spermacocae), but it is otherwise concentrated to the Old World
where it probably originated (32, 33). Subfamily Ixoroideae
shows a similar pattern, because it comprises a species-rich
Neotropical clade (the ‘‘Condaminae–Calycophylleae’’ alliance)
but is otherwise concentrated in the Paleotropics. Contrastingly,
except for tribe Naucleeae, the large subfamily Cinchonoideae is
predominantly Neotropical. In tropical South America, Cinchonoideae is represented by sister tribes Cinchoneae and Isertieae,
which have been shown to build a strongly supported clade (34)
and is sometimes treated as a single tribe (30). Comprising some
130 species of small trees and shrubs divided into 11 genera [see
supporting information (SI) Table S1], the Cinchoneae and
Isertieae are important ecological components of a wide array of
habitats. Some species are also economically important as a
source of quinine. The distribution of Isertieae is concentrated
in the lowlands of the Amazon basin and eastern Guiana,
whereas Cinchoneae species are mainly confined to the highland
and montane habitats of the Northern and Central Andes,
reaching up to 3,300 m (Figs. S1 and S2).
Results and Discussion
Gentry (35), following Raven and Axelrod (36), listed the
Neotropical Rubiaceae as a Gondwana-derived group, evolving
in isolation since the separation of South America from Africa.
This hypothesized origin predicts that (i) the group has a minimal
age of 100 Ma (37), and (ii) Old and New World lineages are
reciprocally monophyletic (6). In contrast, the competing hypothesis of boreotropical origin predicts that (i) South American
groups are derived from northern relatives with Old World taxa
as sister groups, (ii) the divergence between Old and New World
groups occurred between 40 and 50 Ma, i.e., around the Eocene
climatic optimum, which favored the exchange of tropical floristic elements between these land masses (38), and (iii) Early
Tertiary fossils have been found in North America, Europe, or
Asia (6).
Our biogeographic reconstruction (Fig. 1 I and II) corroborates a boreotropical origin in all 3 predictions: (i) our phylogeny
(Fig. S3) shows the Neotropical sister tribes Cinchoneae and
Isertieae nested within a clade of mainly Central American and
Antillean tribes, together sister to the essentially African tribe
Naucleeae and the Paleotropical subfamily Ixoroideae; (ii) our
divergence time estimates place the most recent common ancestor (MRCA) of the Rubiaceae in the Early Paleocene (66.1
Ma, 63.0–68.8; see Table S2), whereas the minimum age of
Cinchonoideae is estimated as only 51.3 (47.8–54.6) Ma, well
after the last known island chain between Africa and South
America possibly existed [Late Cretaceous, ⬇88 Ma (37)]; and
finally, (iii) several Cephalanthus fossils indicate the presence of
the tribe Naucleeae in Europe from the Late Eocene (39) (see
SI Text). Until the Late Eocene or Early Oligocene, a continuous
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belt of boreotropical vegetation covered much of southern North
America, southern Eurasia, and northwestern Africa (40). At
that time, plant migration through direct land connection or
across limited water gaps (6) could have been possible through
the North Atlantic ‘‘Thulean’’ land bridge (13, 41) or through the
Early–Mid Tertiary ‘‘Beringian land bridge’’ (42). Although
climates during the Early Eocene were warmer than today (38),
Beringia was in a considerably higher paleolatitude than the
Thulean bridge. Dispersal of boreotropical elements is therefore
considered more likely across the North Atlantic during this
period (13, 42), which is also supported by the fact that the oldest
Cephalanthus fossils have been found in Europe (39). Together,
these lines of evidence strongly suggest that the Rubiaceae used
the corridors provided by boreotropical vegetation and the
North Atlantic land bridge as a pathway to reach North America
in the Late Paleocene/Early Eocene (Fig. 2I).
From North America, dispersals to South America (Fig. 2II)
may have been facilitated by the proto-Greater Antilles in the
Early Eocene and later by the Greater Antilles and the Avies
Ridge around the Eocene/Oligocene boundary [GAARlandia,
33–35 Ma (43)]. Our divergence time estimates suggest that the
ancestors of Cinchonoideae arrived in northwestern South
America around the Early/Middle Eocene (49.2 Ma, 44.9–53.1;
node 26 in Fig. S4). At that time, sea levels some 50 m above
today’s (44) created a marine incursion from the Caribbean that
limited land dispersal eastwards (1, 24) and another incursion
from the Pacific that blocked dispersal to the south (24, 45, 46)
(Fig. 2III). Lowland areas were covered by closed-canopy tropical rainforests (47). Most of the Andes had not yet been formed,
except for some low mountains in the regions now corresponding
to the Central and Southern Andes (3, 21). Interestingly, the
MRCA of tribes Isertieae and Cinchoneae is reconstructed as
being lowland-adapted (see Fig. S5). But starting in the Middle
Eocene, the Andean orogeny went through a major phase of
mountain building, sometimes referred to as the Incaic II (21,
48). This phase was longitudinally widespread, and in the
northern region it caused uplift of the Central Cordillera (21,
49). The newly formed montane habitats must have acted as an
ecological barrier to lowland taxa, and this seems to explain the
geographic disjunction (Andes vs. Amazonia) between the MRCAs of tribes Cinchoneae and Isertieae (Fig. 1III and Fig. S5).
In the case of Isertieae, their MRCA is most likely reconstructed as being confined to lowland Amazonia (Fig. 2III),
where it first radiated in the Late Oligocene, giving rise to the
genera Kerianthera and Isertia (Fig. 1III and Fig. S5). Diversification in Isertieae occurred mainly in the Middle and Late
Miocene, which is strikingly coincident with the uplift of the
Eastern Cordillera in the Northern Andes [Fig. 1IV; (2, 21)]. For
tribe Cinchoneae, ecological adaptation to higher altitudes
seems to have been the key to its diversification, with most
speciation events confined to montane habitats in the Northern
and Central Andes (Fig. 1 III and IV and Fig. S5).
Western Andean Portal. Most optimizations indicate that the
MRCA of Cinchoneae was confined to the Northern Andes,
presumably in the new habitats created by the Central Cordillera
(Fig. 1III). From the Eocene to the Middle Miocene, some
studies have proposed that marine incursions from the Pacific
invaded a lowland corridor between the Northern and Central
Andes at the latitude of southern Ecuador/northern Peru [⬇3–
5°S (1, 2, 45, 46, 50)], termed the ‘‘Western Andean Portal’’
(WAP, Fig. 2III) or ‘‘Guayaquil Gap’’ (23). Indication for these
incursions comes from fossil occurrences of marine organisms
and palynomorphs (23, 46) and paleosedimentary evidence (1, 2,
50). The WAP is suggested to have been uplifted, and marine
incursions ended, in connection with the uplift of the Eastern
Cordilleras of the Central and Northern Andes from the Middle
Miocene onwards (13–11 Ma) (2, 21, 50).
Antonelli et al.
EVOLUTION
Fig. 1. Combined chronogram and biogeographic analysis of Neotropical Rubiaceae. The tree is the 50% majority-rule consensus (with compatible groups
added) from the Bayesian analysis, with branches proportional to absolute ages (in millions of years) calculated from mean branch lengths of 6,000 Bayesian trees.
Green bars indicate 95% confidence intervals of node ages estimated from 1,000 trees randomly sampled from the Bayesian stationary distribution. Node charts
show the relative probabilities of alternative ancestral distributions obtained by integrating dispersal-vicariance analysis (DIVA) optimizations over the 1,000
Bayesian trees; the first 4 areas with highest probability are colored according to their relative probability in the following order: white ⬎ red ⬎ blue ⬎ gray;
any remaining areas (usually frequencies ⬍0.01) are collectively given with black color. Stars indicate calibration points. Red arrows indicate clades with a
posterior probability ⬍0.90. Present ranges for each species are given after the species name. Brackets identify subfamilies and tribes: CHI, Chiococceae; CIN,
Cinchoneae; GUE, Guettardeae; HAM, Hamelieae; HIL, Hillieae; ISE, Isertieae; NAU, Naucleeae; RON, Rondeletieae. Shaded boxes indicate approximate periods
of Andean uplift phases. The biogeographic interpretation of events I–V is summarized in Fig. 2. (Inset) Areas used in the biogeographic analysis. A, Central
America, B, West Indies; C, Northern Andes; D, Central Andes; E, Chocó; F, Amazonia; G, The Guiana Shield; H, Southeastern South America; I, Temperate North
America; J, Africa; K, Australasia. Topographic map from the National Geophysical Data Center (www.ngdc.noaa.gov).
Antonelli et al.
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Fig. 2. Spatiotemporal evolution of the Neotropical Rubiaceae. (I) Paleocene: Rubiaceae ancestors use the boreotropical route to reach North America from
the Paleotropics. (II) Early Eocene: Dispersal into South America, presumably facilitated by occasional island chains. (III) Late Eocene: North Andean and
Amazonian lineages become isolated by marine incursions such as the Western Andean Portal (WAP). (IV) Middle Miocene: The gradual uplift of the Eastern
Cordillera creates a huge watershed, Lake Pebas (LP). It also closes the WAP, enabling dispersal of plant lineages from the Northern to the Central Andes. (V)
The Pebas system drains, promoting land dispersal of several lineages and rapid speciation of terrestrial plants in western Amazonia. Area codings as in Fig. 1.
(Maps I–II are based on C. R. Scotese’s PALEOMAP project (www.scotese.com); maps III–V modified from refs. 2 and 28).
Our biogeographic reconstruction of subfamily Cinchonoideae (Fig. 1III) corroborates the existence of a dispersal
barrier between the Northern and the Central Andes coincident
with the WAP, and provides indication for the persistence of this
barrier until the Middle Miocene. Throughout the Oligocene
and Early Miocene, all ancestral area reconstructions in tribe
Cinchoneae occur—partly or exclusively—in the Northern
Andes (Fig. 1III). The uplift of the WAP is then observed as at
least 5 independent migrations from the Northern to the Central
Andes within the genera Cinchona and Ladenbergia (Fig. 1IV).
All 5 events are dated as occurring around the Middle/Late
Miocene (12–10 Ma, Fig. S4 and Table S2), almost simultaneously with the suggested end of marine incursions in the WAP
(1, 2, 21, 50).
Several questions remain unanswered concerning the geographic extent and duration of marine settings in the WAP (24,
28). What is clear, however, is that it constituted an important
and long-lasting biogeographic barrier, as evidenced by the fact
that many montane taxa exhibit endemism centers in either side
of the WAP (see SI Text for an expanded discussion), suggesting
that species in those groups were not able to cross the WAP
during their time of radiation. In plants, this pattern has been
recognized in many families, such as Campanulaceae, Calceolariaceae, Tropaeolaceae, Loasaceae, Passifloraceae, Alstroemeriaceae, and Grossulariaceae (see SI Text for references). In
birds, this area has long been recognized as a turnover point
between the Northern and Central Andean biogeographic regions of endemism (51).
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Lake Pebas. Until the end of the Oligocene (⬇24 Ma), a fluvial
system referred to as the paleo-Orinoco dominated the drainage
of northwestern Amazonia and the foreland Andean basins
toward Lake Maracaibo, on the Caribbean coast. Then, in the
Early Miocene (⬇23 Ma) geotectonic changes in the Amazon
Basin associated with the ongoing uplift of the Eastern Cordillera in the Central Andes (28) caused western Amazonia to
gradually become submerged, from south to north and from west
to east. This process created a huge (⬎1 million km2) system of
long-lived lakes and wetlands from at least 17 to 11 Ma, known
as ‘‘Lake Pebas’’ or the ‘‘Pebas Sea’’ (24–28). Whether it was a
purely fluvio-lacustrine (fresh water) system or whether marine
settings were also present is still a matter of debate (24, 27, 28).
However, most authors agree that western Amazonia was completely flooded by some kind of wetland system from at least the
Middle to the Late Miocene and that this system was connected
to the Caribbean marine incursion in the north (Fig. 2IV). From
the Late Miocene onwards (11–7 Ma), there was a new period
of rapid mountain uplift, affecting mainly the Eastern Andean
Cordilleras [sometimes termed Quechua phases II and III (21,
49)]. This presumably caused the western margin of the Guiana
Shield to emerge, which closed the Caribbean connection of the
paleo-Orinoco, shifted the drainage of the Amazon Basin eastwards, and lead to the demise of Lake Pebas (27, 28) (Fig. 2V).
Aquatic conditions, however, seem to have persisted in westerncentral Amazonia until at least 7 Ma, when the modern Amazon
system came into place (28).
Previous studies have provided indications to the potential
role played by Lake Pebas as a pathway for marine organisms to
Antonelli et al.
Materials and Methods
Phylogenetic analyses were performed under parsimony and Bayesian methods as implemented in PAUP* (53) and MrBayes3 (54). We used representative
genera of 4 families of Gentianales as outgroup, based on evidence that the
Rubiaceae are the sister group to the rest of Gentianales (e.g., ref. 55). The
ingroup included representatives of all Rubiaceae subfamilies, with focus on
subfamily Cinchonoideae in which all tribes were represented (Fig. 1). The
final dataset comprised 62 species and 5,894 characters, derived from matK,
rbcL, ITS1–5.8S–ITS2, trnL-F, and rps16 (Table S3). Thus, our dataset comprised
some 3–3.5 times more characters than similar studies (e.g., refs. 9 and 19),
which resulted in a robust phylogeny where 80% of all tree nodes were
strongly supported (Bayesian posterior probability values, pp ⱖ0.95 or
jackknife ⱖ85%). Dataset and trees are available from TreeBase (www.
treebase.org) accession nos. S2334 and M4437. Evolutionary rates were estimated with the Penalized Likelihood algorithm implemented in r8s (56).
Accurate fossil calibration has been suggested as a key factor in age estimation
(57). Here, absolute ages were estimated by using fossil fruits and seeds of
Cephalanthus, a genus with an exceptionally rich and reliable fossil record
from the Late Eocene onwards (Fig. S4). Additionally, a maximum age constraint of 78 Ma was independently enforced on the basal node of the tree
1. Hoorn C (1993) Marine incursions and the influence of Andean tectonics on the
Miocene depositional history of northwestern Amazonia: Results of a palynostratigraphic study. Palaeogeogr Palaeocl 105:267–309.
2. Hoorn C, Guerrero J, Sarmiento GA, Lorente MA (1995) Andean tectonics as a cause
of changing drainage patterns in Miocene northern South America. Geology
23:237–240.
3. Gregory-Wodzicki KM (2000) Uplift history of the central and northern Andes: A
review. Geol Soc Am Bull 112:1091–1105.
4. Myers N, Mittermeier RA, Mittermeier CG, da Fonseca GAB, Kent J (2000) Biodiversity
hotspots for conservation priorities. Nature 403:853– 858.
5. Kreft H, Jetz W (2007) Global patterns and determinants of vascular plant diversity.
Proc Natl Acad Sci USA 104:5925–5930.
6. Pennington RT, Dick CW (2004) The role of immigrants in the assembly of the South
American rainforest tree flora. Philos Trans R Soc London Ser B 359:1611–1622.
7. Donoghue MJ (2008) A phylogenetic perspective on the distribution of plant diversity.
Proc Natl Acad Sci USA 105:11549 –11555.
Antonelli et al.
based on the crown age of Gentianales (58). Finally, to incorporate topological
and branch length uncertainty in our age estimates, 1,000 trees randomly
sampled from the Bayesian stationary distribution were independently dated
and results summarized to obtain median values and 95% credibility intervals
of node ages (Fig. S4 and Table S2). See SI Text for a detailed description of the
phylogenetic and dating methods used.
Eleven areas (Fig. 1 Inset) were defined for the biogeographic analysis
based on the extant distribution patterns of Rubiaceae and on geological
history (3, 21). Whenever possible, we tried to maximize congruence with
other biogeographic studies in South America (59, 60). Dispersal-vicariance
analysis [DIVA (61, 62)] was used to infer ancestral distributions and historical
events involved in the biogeographic history of Rubiaceae. To overcome the
uncertainty associated with phylogenetic estimation, we use an approach that
averages DIVA biogeographic and temporal reconstructions over a Bayesian
sample of highly probable trees (in this case n ⫽ 1,000), generating credibility
support values for alternative phylogenetic relationships (63). Integrating
over the posterior distribution of trees often reveals preference for a single or
more restricted set of solutions, thus reducing the uncertainty in DIVA optimizations (63).
Incomplete taxon sampling may cause problems for historical inference
methods because it reduces the accuracy of ancestral state reconstructions
(see ref. 64 for a different view). This is particularly problematic in a large
family such as Rubiaceae, spanning nearly all continents. To overcome this
problem, we used encompassing areas outside the Neotropics, where most of
Ixoroideae (as well as the first diverging clade in Cinchonoideae, the tribe
Naucleeae) are found. We also estimated the geographic bias in our sample
for the Neotropical tribes Cinchoneae and Isertieae, the main focus of this
article. Results showed that our taxon sampling is fairly representative of the
actual distribution of the species within each genus and tribe (see Table S4, Fig.
S6, and SI Text for a detailed discussion of distribution patterns in these tribes).
In addition, to test the sensitivity of our DIVA reconstructions to missing taxa,
we performed a series of heuristic simulations in which we added hypothetical
taxa from underrepresented areas to key nodes in the phylogeny of these 2
tribes. DIVA ancestral reconstructions proved to be very stable to the addition
of missing taxa, at least for the nodes involved in the WAP and Lake Pebas
scenario (Fig. S7). We also examined the effect of missing taxa on divergence
time estimations by randomly deleting taxa from the original sample and
calculating divergence times on these reduced datasets. As Linder et al. (65)
suggested, PL proved to be largely insensitive to taxon sampling, and for most
nodes the estimated ages were within the 95% confidence interval from the
complete dataset (Fig. S8; see SI Text for more details on simulations). This
gives us confidence that the biogeographic scenario presented here (Fig. 2)
would not be significantly altered by the addition of missing taxa.
ACKNOWLEDGMENTS. We are indebted to F. Wesselingh, C. Hoorn, R. Eriksson, E. M. Friis, P. Endress, S. Schultka, B. Bremer, C. Rydin, E. Kowalski, T.
Sempere, L. Kinoshita, K. Yamamoto, T. Eriksson, M. Sanderson, K. Suguio, R.
Bemerguy, M. Pirie, O. Seberg, J. Ohlson, B. Oxelman, and N. Wikström for
invaluable advice and to V. Aldén for technical assistance. This manuscript has
been greatly improved thanks to the suggestions of 2 anonymous reviewers
and the editor. We wish to dedicate this study to the memory of Lennart
Andersson, who initiated the project and participated actively in it until his
death in January 2005. This work was supported by grants from the Swedish
Research Council (to C. P.), from the Royal Swedish Academy of Sciences, the
Royal Society of Arts and Sciences in Gothenburg, Kungliga och Hvitfeldtska
Stiftelsen, Carl Tryggers Stiftelse, Helge Ax:son Johnsons Stiftelse, and University of Gothenburg (to A.A.), and from the Program ‘‘Ramon y Cajal’’ of the
Spanish Ministry of Education and Science and the ‘‘Biogeography Working
Group’’ supported by NESCent (National Science Foundation Grant EF0423641) (to I.S.).
8. Wiens JJ, Donoghue MJ (2004) Historical biogeography, ecology, and species richness.
Trends Ecol Evol 19:639 – 644.
9. Davis CC, Bell CD, Mathews S, Donoghue MJ (2002) Laurasian migration explains
Gondwanan disjunctions: Evidence from Malpighiaceae. Proc Natl Acad Sci USA
99:6833– 6837.
10. Richardson JE, Chatrou LW, Mols JB, Erkens RHJ, Pirie MD (2004) Historical biogeography of two cosmopolitan families of flowering plants: Annonaceae and Rhamnaceae. Philos Trans R Soc London Ser B 359:1495–1508.
11. Pirie MD, Chatrou LW, Mols JB, Erkens RHJ, Oosterhof J (2006) Andean-centred genera
in the short-branch clade of Annonaceae: Testing biogeographical hypothesis using
phylogeny reconstruction and molecular dating. J Biogeogr 33:31– 46.
12. Tiffney BH (1985a) Perspectives on the origin of the floristic similarity between eastern
Asia and eastern North America. J Arnold Arbor 66:73–94.
13. Tiffney BH (1985b) The Eocene North Atlantic land bridge: Its importance in Tertiary
and modern phytogeography of the Northern Hemisphere. J Arnold Arbor 66:243–
273.
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EVOLUTION
disperse into fresh water biotopes, based on fossil occurrences of
mollusks and DNA phylogenies of Neotropical fishes (23, 24, 26).
On the other hand, if this wetland system was as large and
interconnected as suggested, it should also have acted as a biotic
barrier to the dispersal of terrestrial organisms between the
Andes and the eastern Amazonian and Guiana regions.
Indeed, the existence of a dispersal barrier between the Andes
and lowland Amazonia during the Middle Miocene is corroborated by our biogeographic reconstructions (Fig. 1IV). Until the
Early/Middle Miocene boundary, several ancestors in tribe
Cinchoneae are inferred to have been widespread in both of
those areas. But after that—coinciding with the time Lake Pebas
is proposed to have existed—all ancestral lineages in Cinchoneae
and Isertieae appear to have occupied either the Andes or
Amazonia, but not both. Most endemic species of Isertieae are
currently found in Guiana and eastern Amazonian lowlands (Fig.
S1 A), whereas western-central Amazonian distributions are
mainly represented by widespread species, suggesting that these
distributions are the result of recent range expansion, probably
after the drying of Lake Pebas. Similarly, several ancestral nodes
in Remijia are reconstructed as occurring on both sides of the
barrier (Fig. 1V), but these nodes are dated after the Miocene/
Pliocene boundary (⬇5.3 Ma, Fig. S4) and thus postdate the
closing of the Pebas wetland system. The final reestablishment
of land connections between the Andes and Amazonia is evidenced by at least 7 independent colonization events in Cinchoneae and Isertieae from the Late Miocene onwards (Fig. 1V).
Eventually, the emergence of new lands in Central America after
the closing of the Panama Isthmus (3.5 Ma) (21, 52) provided
suitable areas for northwards dispersal of South American
lineages. Species such as Joosia umbellifera and Isertia haenkeana,
which are now widespread in Central America, probably
dispersed soon after the establishment of that land connection
(Fig. 2V).
14. Wolfe JA (1975) Some aspects of plant geography of the Northern Hemisphere during
the Late Cretaceous and Tertiary. Ann Mo Bot Gard 62:264 –279.
15. Moore BR, Donoghue MJ (2007) Correlates of diversification in the plant clade Dipsacales: Geographic movements and evolutionary innovations. Am Nat 170:S28 –S55.
16. Bell CD, Donoghue MJ (2005) Phylogeny and biogeography of Valerianaceae (Dipsacales) with special reference to the South American valerians. Organisms Divers Evol
5:147–159.
17. Winkworth RC, Donoghue MJ (2005) Viburnum phylogeny based on combined molecular data: Implications for taxonomy and biogeography. Am J Bot 92:653– 666.
18. von Hagen KB, Kadereit JW (2003) The diversification of Halenia (Gentianaceae):
Ecological opportunity versus key innovation. Evolution (Lawrence, Kans) 57:2507–
2518.
19. Hughes C, Eastwood R (2006) Island radiation on a continental scale: Exceptional rates
of plant diversification after uplift of the Andes. Proc Natl Acad Sci USA 103:10334 –
10339.
20. Brumfield RT, Edwards SV (2007) Evolution into and out of the Andes: A Bayesian
analysis of historical diversification in Thamnophilus ant shrikes. Evolution (Lawrence,
Kans) 61:346 –367.
21. Taylor DW (1991) Paleobiogeographic relationships of Andean angiosperms of Cretaceous to Pliocene age. Palaeogeogr Palaeocl Palaeoecol 88:69 – 84.
22. Garzione CN, et al. (2008) Rise of the Andes. Science 320:1304 –1307.
23. Nuttal CP (1990) A review of the Tertiary non-marine molluscan faunas of the Pebasian
and other inland basins of north-western South America. Bull Brit Mus Nat Hist Geol
45:165–371.
24. Lundberg JG, et al. (1998) in Phylogeny and Classification of Neotropical Fishes, eds
Malabarba LR, Reis RE, Vari RP, Lucena ZM, Lucena CAS (Edipucrs, Porto Alegre, Brazil),
pp 13– 48.
25. Wesselingh FP, et al. (2002) Lake Pebas: A palaeoecological reconstruction of a Miocene, long-lived lake complex in western Amazonia. Cainozoic Res 1:35– 81.
26. Wesselingh FP (2006) Miocene long-lived lake Pebas as a stage of mollusc radiations,
with implications for landscape evolution in western Amazonia. Scripta Geol 133:1–17.
27. Wesselingh FP, Salo JA (2006) A Miocene perspective on the evolution of the Amazonian biota. Scripta Geol 133:439 – 458.
28. Wesselingh FP, et al. (2010) in Amazonia: Landscape and species evolution, eds Hoorn
C, Wesselingh FP (Blackwell Publishing, Oxford), in press.
29. Lovejoy NR, Albert JS, Crampton WGR (2006) Miocene marine incursions and marine/
freshwater transitions: Evidence from Neotropical fishes. J S Am Earth Sci 21:5–13.
30. Robbrecht E, Manen JF (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.
31. Govaerts R, Frodin DG, Ruhsam M, Bridson DM, Davis AP (2007) World Checklist and
Bibliography of Rubiaceae (The Trustees of the Royal Botanic Gardens, Kew, UK).
32. Manen JF, Dessein S, De Block P, Robbrecht E (2006) Towards an evolutionary scenario
for the coffee family (Rubiaceae, angiosperms): Further insights from a supertree.
Scripta Bot Belg 40:47.
33. Sonké B, Dessein S, Taedoumg H, Groeninckx I, Robbrecht E (2008) A new species of
Colletoecema (Rubiaceae) from southern Cameroon with a discussion of relationships
among basal Rubioideae. Blumea 53:533–547.
34. Andersson L, Antonelli A (2005) Phylogeny of the tribe Cinchoneae (Rubiaceae), its
position in Cinchonoideae, and description of a new genus, Ciliosemina. Taxon 54:17–
28.
35. Gentry AH (1982) Neotropical floristic diversity: Phytogeographical connections between Central and South America, Pleistocene climatic fluctuations, or an accident of
the Andean orogeny? Ann Mo Bot Gard 69:557–593.
36. Raven PH, Axelrod DI (1974) Angiosperm biogeography and past continental movements. Ann Mo Bot Gard 61:539 – 673.
37. Morley RJ (2003) Interplate dispersal routes for megathermal angiosperms. Perspec
Plant Ecol Evol Syst 6:5–20.
6 of 6 兩 www.pnas.org兾cgi兾doi兾10.1073兾pnas.0811421106
38. Zachos J, Pagani M, Sloan L, Thomas E, Billups K (2001) Trends, rhythms, and aberrations in global climate 65 Ma to the present. Science 292:686 – 693.
39. Mai DH, Walther H (1985) The Upper Eocene floras of the Weisselster basin and
adjacent areas. (in German) Abh Staat Mus Miner Geol Dresden 33:1–260.
40. Lavin M, Luckow M (1993) Origins and relationships of tropical North America in the
context of the boreotropics hypothesis. Am J Bot 80:1–14.
41. McKenna MC (1983) in Structure and Development of the Greenland-Scotland Bridge:
New Concepts and Methods, eds Bott MHP, Saxov S, Talwani M, Thiede J (Plenum, New
York), pp 351–395.
42. Sanmartı́n I, Enghoff H, Ronquist F (2001) Patterns of animal dispersal, vicariance and
diversification in the Holarctic. Biol J Linnean Soc 73:345–390.
43. Iturralde-Vinent MA, MacPhee RDE (1999) Paleogeography of the Caribbean region:
Implications for Cenozoic biogeography. Bull Am Mus Nat Hist 238:1–95.
44. Miller KG, et al. (2005) The phanerozoic record of global sea-level change. Science
310:1293–1298.
45. Steinmann M, Hungerbühler D, Seward D, Winkler W (1999) Neogene tectonic evolution and exhumation of the southern Ecuadorian Andes; a combined stratigraphy and
fission-track approach. Tectonophysics 307:255–276.
46. Santos C, Jaramillo C, Bayona G, Rueda M, Torres V (2008) Late Eocene marine incursion
in north-western South America. Palaeogeogr Palaeocl 264:140 –146.
47. Morley RJ (2000) Origin and Evolution of Tropical Rain Forests (Wiley, Chichester, UK).
48. Noble DC, McKee EH, Mourier T, Mégard F (1990) Cenozoic stratigraphy, magmatic
activity, compressive deformation, and uplift in northern Peru. Bull Geol Soc Am
102:1105–1113.
49. Irving EM (1975) Structural evolution for the northernmost Andes, Colombia. USGS
846:1– 47.
50. Hungerbühler D, et al. (2002) Neogene stratigraphy and Andean geodynamics of
southern Ecuador. Earth Sci Rev 57:75–124.
51. Stotz DF, Fitzpatrick JW, Parker TA, Moskovits DK (1996) Neotropical Birds: Ecology and
Conservation (University of Chicago Press, Chicago).
52. Briggs JC (1994) The genesis of Central America: Biology versus geophysics. Glob Ecol
Biogeogr 4:169 –172.
53. Swofford DL (2002) PAUP*. Phylogenetic Analysis Using Parsimony (*and other
methods) (Sinauer, Sunderland, MA), Ver 4.0b10.
54. Ronquist F, Huelsenbeck JP (2003) MrBayes3: Bayesian phylogenetic inference under
mixed models. Bioinformatics 19:1572–1574.
55. Albach DC, Soltis PS, Soltis DE, Olmstead RG (2001) Phylogenetic analysis of asterids
based on sequences of four genes. Ann Mo Bot Gard 88:163–212.
56. Sanderson MJ (2002) Estimating absolute rates of molecular evolution and divergence
times: A penalized likelihood approach. Mol Biol Evol 19:101–109.
57. Sauquet H, et al. (2009) Contrasted patterns of hyperdiversification in Mediterranean
hotspots. Proc Natl Acad Sci USA 106:221–225.
58. Bremer K, Friis EM, Bremer B (2004) Molecular phylogenetic dating of asterid flowering
plants shows early Cretaceous diversification. Syst Bot 53:496 –505.
59. Cracraft J (1988) Deep-history biogeography, retrieving the historical pattern of
evolving continental biotas. Syst Zool 37:221–236.
60. Posadas PE, Estévez JM, Morrone JJ (1997) Distributional patterns and endemism areas
of vascular plants in the Andean subregion. Fontqueria 48:1–9.
61. Ronquist F (1996) DIVA version 1.1. Computer program and manual available from
Uppsala University (www.ebc.uu.se/systzoo/research/diva/diva.html).
62. Ronquist F (1997) Dispersal-vicariance analysis: A new approach to the quantification
of historical biogeography. Syst Biol 46:195–203.
63. Nylander JAA, Olsson U, Alström P, Sanmartı́n I (2008) Accounting for phylogenetic
uncertainty in biogeography: A Bayesian approach to dispersal-vicariance analysis of
the Thrushes (Aves: Turdus). Syst Biol 57:257–268.
64. Li G, Steel M, Zhang L (2008) More taxa are not necessarily better for the reconstruction
of ancestral character states. Syst Biol 57:647– 653.
65. Linder HP, Hardy CR, Rutschmann F (2005) Taxon sampling effects in molecular clock
dating: An example from the African Restionaceae. Mol Phylogen Evol 35:569 –582.
Antonelli et al.
Supporting Information
Antonelli et al. 10.1073/pnas.0811421106
SI Text
Molecular dating. Divergence times were estimated on the tree
Expanded Materials and Methods. Phylogenetic analyses. We recon-
topology with the highest posterior probability from the Bayesian analysis, but using mean branch lengths calculated from 1,000
post-burn-in trees from the Bayesian sample (see below). A
likelihood ratio test (13) strongly rejected the hypothesis of a
molecular clock for the tree (P ⬍ 0.0001). The clockindependent algorithm Penalized Likelihood (14) implemented
in the software r8s v. 1.70 (15) was then used for estimating
divergence times in the tree.
Fossils have been assigned to some 10 genera of extant
Rubiaceae in our phylogeny (ref. 17; the Paleobiology Database,
http://paleodb.org). However, in the majority of cases, a careful
evaluation of the original fossil descriptions made evident that
almost none of these could be deemed fully reliable in terms of
taxonomic placement and/or geologic age. Approximately more
than half of these records are fragmentary traces of leaves, and
the rest are pollen grains examined with light microscopy.
Considering the great convergence of leaf and pollen forms
among angiosperms, which render even fresh material very
difficult to identify in the absence of various other vegetative and
floral parts, we have therefore opted for not using any of these
2 types of fossils as calibration points.
An important exception among Rubiaceae fossils exists in
Cephalanthus. The oldest fossil of the genus is Cephalanthus
kireevskianus from the Late Eocene of Germany (18), which
appears to be both correctly identified (E. M. Friis, personal
communication) and dated (S. Schultka, personal communication). Additional findings indicate that this species was common
and widespread from the Late Eocene onwards in Europe (19)
and in Western Siberia, where the species was originally described (20, 21). Another fossil species of the genus, C. pusillus,
was described from the Middle Miocene of Denmark (22). Fossil
fruits of Cephalanthus possess several morphological features
that make them taxonomically recognizable by means of overall
similarity. These include fruit type, a schizocarp; mericarp
obovoid, slightly dorsoventrally flattened; ventral face flat with
a shallow median furrow; dorsal face convex, apically truncate;
placentation apical; seed strophiolate, apotropous pendulous,
obovoid, slightly dorsoventrally flattened; strophiole apical, sickle-shaped; hilum dorsal, marked by a narrow slit (ref. 22; Fig.
S4A Inset).
The continuous fossil records in every single geologic epoch
from the Late Eocene to the Pliocene in almost 20 fossil sites
(Fig. S4 B and C), combined with the reliability of their
taxonomic placement thanks to a high quality of preservation,
make Cephalanthus an ideal calibration point for the Rubiaceae
phylogeny. The oldest fossil finding in Cephalanthus was therefore used to place a minimum age constraint of 33.9 Ma on the
first split in tribe Naucleeae, separating Cephalanthus and Hallea
⫹ Nauclea. The stratigraphical age of this fossil was converted
to an absolute age by using the ending point of the geological
epoch to which it was assigned, using the time scale of Gradstein
et al. (23). In addition, a maximum age constraint of 78 Ma was
independently enforced for the basal node in the tree based on
the crown age of Gentianales, as estimated from a well-sampled
analysis of the asterids calibrated with multiple fossils (16).
Although our approach of using a single fossil calibration may
seem too conservative (it could be that some other Rubiaceae
fossils are correctly identified and dated) most fossils described
in the literature (including the Posoqueria and Cosmibuena
fossils cited in ref. 17) are too young to produce any influence
in the molecular dating results obtained by using the Cephalan-
structed phylogenetic relationships using DNA sequences from
5 different markers: the matK and rbcL genes, the ITS1-5.8S–
ITS2 and trnL–F regions, and the rps16 intron. Most of these
sequences were previously obtained by using the methodology
published in Andersson and Antonelli (1), except for 6 additional species of Isertia (I. haenkeana, I. hypoleuca, I. parviflora,
I. pittieri, I. rosea, and I. spiciformis) that were newly sequenced
for this study to increase taxon sampling within tribe Isertieae.
Accession numbers and source of all sequences included in this
study are listed in Table S3. Following Andersson and Antonelli
(1), we used representative genera of 4 families of Gentianales
as outgroup for the analysis, based on evidence that the Rubiaceae are the sister group to the rest of Gentianales (e.g., ref. 2).
The dataset also included representatives from the 2 other
Rubiaceae subfamilies Rubioideae and Ixoroideae, as well as
genera representing all tribes in subfamily Cinchonoideae (Fig.
1). In addition, an extra outgroup species (Nicotiana tabacum)
was included in the phylogenetic analysis to determine the
position of the root within the basal branch, as required by the
molecular dating method described below, but this taxon was
subsequently excluded from the results. The complete dataset
was realigned by using MAFFT v. 5.64 (3) and adjusted manually. Gaps were coded as present/absent (0/1) following the
principles outlined by Antonelli (4) and included in the phylogenetic analysis. The final aligned data matrix comprised 5,894
characters, of which 1,600 derived from matK, 1,398 from rbcL,
1,254 from trnL-F, 908 from rps16, 676 from the ITS region, and
58 from gap codings. Of 2,153 variable characters, 1,165 were
parsimony informative.
Parsimony jackknife support values (5) were estimated in
PAUP v. 4.0b10 (6) by running 10,000 replicates with 37%
deletion, 10 random addition sequence replicates, using TBR
branch swapping and saving up to 30 trees per replicate. Following recent works (7, 8), the Akaike Information Criterion,
implemented in MrModelTest v. 2.2 (9), was used to choose the
optimal model of sequence evolution for each DNA marker.
Bayesian analyses were subsequently implemented in MrBayes v.
3.1.2 (10) using the GTR⫹G model for matK and rps16, and the
GTR⫹I⫹G model for ITS, rbcL, and trnL-F. Gap-codings were
analyzed as a separate partition under the Restriction Site
(Binary) Model. Two simultaneous analyses with 8 MetropolisCoupled MCMC chains each were run for 2.5 million generations, sampling every 500th generation. Each analysis was started
from different, randomly sampled topologies and let to run until
the average standard deviation of split frequencies became
⬍0.01, indicating convergence of trees and model parameters
across the runs (11). The burn-in value was set in all cases to
1,000,000 generations after visual inspection of the split (clade)
frequencies using the software AWTY (12). Final results were
based on the pooled values from the 2 independent analyses. The
parsimony jackknife consensus tree was virtually congruent with
the Bayesian tree (Fig. S3), and both were very similar to the tree
of Andersson and Antonelli (1). Nearly 80% of all tree nodes
were strongly supported (Bayesian posterior probability values,
pp ⱖ95% or jackknife ⱖ85%). Kerianthera was placed with
strong support as the sister to Isertia, and within the latter there
were 3 strongly supported clades of sister species (Fig. S3). The
Bayesian and jackknife trees, as well as the aligned dataset used
for the analyses, are available from TreeBase (www.treebase.
org), accession numbers S2334 and M4437.
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
1 of 20
thus and root calibrations (Fig. S4). The only potential exceptions
are the following 2 fossils, which may have been correctly dated, but
that we do not consider reliable for the reasons specified:
1) Assigned name: Remijia tenuiflorafolia
Type of fossil: Leaf traces
Minimum age: 47.46
Country: Argentina
Primary reference: ref. 24.
Comments: In our judgment, the leaf characters originally
used for placing these fossils in Remijia (size 9.5 ⫻ 2.25 cm,
secondary veins 10–13, camptodromous venation, equally acute
apex and base), as well as the illustrations provided, indicate that
these fossil leaves could equally well belong to several other
species of Rubiaceae (e.g., Agouticarpa curviflora, Kutchubaea
surinamensis, or Alibertia bertierifolia, in tribe Gardenieae) or
even to other plant families. Moreover, this taxon was described
from southwestern Argentina, which is some 3,000 km south of
the southernmost border of the present-day range of the genus
(Fig. S1D).
2) Assigned name: Psychotria eogenica
Type of fossil: Seeds
Minimum age: 28.4
Country: Peru
Primary reference: ref. 25.
Comments: As traditionally circumscribed, Psychotria is one of
the most species-rich genera among angiosperms, at most comprising some 1,950 species (26, 27). Phylogenetic analyses have
shown, however, that ‘‘Psychotria is broadly paraphyletic and
defined by lack of characters used to define other genera in the
tribe’’ (26). It is thus not surprising that the placement of this
fossil in Psychotria was originally done ‘‘with some hesitation for
the reason that in all seeds of that genus that I have seen they
are smaller than the fossils and the ribbing is more continuous
and is also more pronounced, as is also true in the case of related
genera such as Phialanthus Grisebach and Ixora Linné’’ (25).
All other fossils listed in the Paleobiology Database and the
literature for the genera sampled in this study would have been
far too recent to reach the lower bound of the age intervals
calculated using the 2 calibration points we rely on (i.e., Cephalanthus and the root). In sum, we argue that it is a better
approach to base our dating analysis on a single (but by all means
reliable) fossil, than using several more or less dubious records.
Finally, to account for topological and branch length uncertainty in our age estimates, 1,000 randomly chosen trees from the
Bayesian stationary sample were independently dated and results summarized to obtain the median value and 95% credibility
intervals of node ages (Fig. S4 and Table S2), by using the
softwares TreeAnnotator and FigTree v. 1.4 (28).
Biogeographic Analysis. Distribution and altitudinal data. Except for
Remijia, all larger genera of Cinchoneae and Isertieae have been
the subject of recent taxonomic revisions (29–32), and fairly
detailed distribution data are available for all major taxa (29–
39). Distribution data on the remaining Neotropical Rubiaceae
and the paleotropical genera were obtained from Andersson
(40), Bridson and Verdcourt (41), Mabberley (42), Puff (43), and
Smith (44). In addition, we searched for recent collections
retrieved from various herbaria at the Global Biodiversity Information Facility (www.gbif.org), but such datapoints were only
used when they were deemed reliable (e.g., identifications by
experts).
Operational areas used in the biogeographic analysis. The delimitation
of areas for the biogeographic analysis was based on the extant
distribution patterns of Rubiaceae taxa (i.e., congruent distributional ranges shared by 2 or more species) and on geological
history, i.e., areas historically isolated from one another by
dispersal barriers (45, 46). In South America, we also tried to
maximize congruence with other biogeographic studies in the
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
region by selecting similarly defined areas (47–50). However, our
altitudinal boundaries were usually lower than it is commonly
adopted, reflecting ecological constraints in our taxon distributions (many Rubiaceae are premontane species). In all, 11 areas
were defined:
A: Central America. From southern Mexico (Veracruz, Oaxaca,
Tabasco, Campeche, Yucatán and Quintana Roo) south to
Panama. Although this region has had a complex geologic
history and its land and island connections to South America are
still prone to discussion, there has been a long-term isolation
from South America until the uplift of the Panama Isthmus at 3.5
Ma (45, 51).
B: West Indies. Excluding Trinidad and Tobago, which are
geologically and biologically more closely related to South
America than to the other Caribbean islands. Although the
Greater and the Lesser Antilles have different geologic histories,
they have been long isolated by water from other American
landmasses.
C: Northern Andes (10° N–5° S). From Venezuela and Colombia
south to northernmost Peru (Piura, Cajamarca, and Amazonas),
from elevations ⬎500 m. This area is roughly the same as the
Páramo recognized in other biogeographic studies (e.g., refs.
48–50), except that occurrences in this area are arbitrarily coded
beginning at altitudes somewhat inferior to the ones generally
adopted (due to the distributional patterns shown by our study
taxa). Our delimitation of the Northern Andes is slightly different from that of Taylor (45) ranging from 10°N to 3°S. The
reason for this is that we wanted the boundary between the
Northern and the Central Andes to coincide with the Western
Andean Portal (Guayaquil Gap), following Hoorn (52) and
Hurgerbühler et al. (53). This region seems to be a major
biogeographic barrier for many groups of Neotropical plants (see
ref. 54). Also, many Cinchoneae species endemic to area C
extend their range down to 5–6°S.
D: Central Andes (5°S–18°S). From Peru (San Martin and La
Libertad) southwards to the Tropic of Capricorn, from elevations higher than 500 m. Similarly to the operational area defined
for the Northern Andes, this area approximately corresponds to
the Puna or Altiplano commonly recognized (e.g., refs. 48–50)
but is used here as if occurring from a lower altitudinal limit.
E: The Chocó area. Comprises areas west of the Andes and
below 500-m altitude in Colombia (Chocó, El Valle, Cauca, and
Nariño), Ecuador, and Peru (Tumbes, Piura). This area is usually
recognized by bird biogeographers as a center of endemism (e.g.,
refs. 47 and 55).
F: Amazonia. Comprises the lowland (⬍500 m) vegetation in
Colombia, Ecuador, Peru, Bolivia, Brazil, Venezuela, Guyana,
Suriname, and French Guiana and includes the islands immediately off the South American coast.
G: The Guiana Shield. Includes the elevated (ⱖ500 m) areas in
northeastern South America, comprising parts of Venezuela,
Guyana, Suriname, French Guiana, and Brazil. It corresponds to
the Guianan Bedrock region.
H: Southeastern South America. Mostly comprised of the Brazilian Shield, but also including the lowlands in eastern Brazil
and the Rio Paraná drainage. The area corresponds to the
pre-Cambrian Brazilian Bedrock formation.
I: Temperate North America. From the Tropic of Cancer northwards. This area is ecologically separated from Central America
by the current occurrence of arid and semiarid habitats in
northern Mexico.
J: Africa. Separated from South America at about 110–100 Ma
(56, 57).
K: Australasia. Includes also Madagascar and the Seychelles,
which were separated from Africa together with India already at
⬇121 Ma (58).
Dispersal-vicariance analysis. Dispersal-vicariance analysis (DIVA,
refs. 59 and 60) was used to infer ancestral distributions at
2 of 20
internal nodes in the phylogeny of Rubiaceae and to identify the
biogeographic events involved in the history of the group. DIVA
has the advantage over more cladistically oriented methods that
it makes no a priori assumptions about the shape or existence of
general biogeographic patterns, making it very useful in regions
where area relationships have varied greatly through time such
as the Neotropics (47). However, like most biogeographic inference methods (61, 62), DIVA requires completely resolved, fully
bifurcated trees. This can be a problem because unresolved trees
are a common feature of Andean radiations, presumably due to
rapid diversification rates. To account for phylogenetic uncertainty in our biogeographic reconstructions, we used here a
method that averages DIVA reconstructions over a Bayesian
sample of trees (in this case n ⫽ 1,000) reflecting credibility
values on each clade (63). Moreover, it has been observed that
integrating over the posterior distribution of trees often reveals
preference for a single or more restricted set of solutions, thus
reducing the uncertainty in DIVA optimizations (63). DIVA
analyses were run unconstrained, i.e., with no constraint in the
maximum number of areas applied to ancestral area optimizations (59).
Altitudinal optimization. To infer ancestral altitudinal ranges, Maximum Parsimony (Fitch) optimizations were performed in the
software Mesquite v. 2.0.1 (64). Independent optimizations were
run on 5,000 Bayesian trees from a stationary tree sample and
plotted on the 50% majority-rule consensus tree (compatible
groups added) of the Bayesian analysis.
Expanded Results and Discussion. Diversity patterns in Neotropical
Rubiaceae show that although it is cosmopolitan in distribution,
the highest diversity in family Rubiaceae is distinctly confined to
the tropics. Subfamily Rubioideae is pantropically distributed
and comprises some highly diverse groups in the Neotropics
(e.g., Palicoureeae and Spermacoceae) but is otherwise concentrated to the Old World where it probably originated (65, 66).
The majority of species in the subfamily Ixoroideae (except for
the ‘‘Condamineeae–Calycophylleae’’ alliance) are also found in
the Paleotropics, whereas the subfamily Cinchonoideae, with the
exception of tribe Naucleeae, is predominantly Neotropical.
There are several tribes (Hamelieae, Hillieae, Chiococceae,
Rondeletieae, and Guettardeae) distributed in Central America
and the West Indies, where they inhabit wet lowlands and cloud
forests, with some species extending their range to southern
North America, and others (such as in the genus Hillia) confined
to South America.
Cinchonoideae is represented in tropical South America by
the sister tribes Cinchoneae and Isertieae (1). A list of all
currently recognized genera is shown in Table S1 and their
species listed in Table S4. Figs. S1–S2 show detailed distribution
maps for the main genera in Cinchoneae and Isertieae still not
depicted by earlier studies (drawn by Lennart Andersson).
The tribe Isertieae is more or less continuously distributed
from Guatemala to central Bolivia and the Amazon mouth (Fig.
1 A). The greatest species diversity (3–4 species per grid square)
is found in eastern Guiana and adjacent parts of the Amazon
basin, between 68°W and 74°W, but there is not a very pronounced center of diversity (the highest number of species per
grid square is only 5). The range west of 72°W is occupied by I.
haenkeana, I. rosea, and species of the subgenus Cassupa, which
do not occur east of 66°W. The high species richness in the
central Amazon basin is mainly due to the overlap of the ranges
of widespread species, whereas the species richness in eastern
Guiana and easternmost Amazonia depends mainly on the
presence of narrowly restricted endemics.
The distribution range of Cinchoneae (Fig. S1B) comprises
most of the Neotropical region. Within this range, two distinct
centers of diversity can be found, one along the Andes and
another in central Amazonia. The Andean center is formed
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
mainly by species of Cinchona, Joosia, and Ladenbergia, whereas
the eastern Amazonian center comprises a large number of
endemic Remijia species. The genera Ciliosemina, Cinchonopsis,
Maguireocharis, and Pimentelia have too few species to clearly
show any center of diversity. Cinchoneae is also present in the
Brazilian Shield through two species of Ladenbergia (L. chapadensis and L. cujabensis). There is one species of Remijia (R.
ferruginea) isolated on the southeastern fringe of the Brazilian
Shield as well as scattered occurrences of Ladenbergia hexandra
in the Atlantic coast of southeastern Brazil.
The distribution range of Ladenbergia (Fig. S1C) covers most
of the range of Cinchoneae, but species diversity is highest in the
Andes. Most species have relatively restricted ranges (31), with
only a few species extending further than 10° in direction north
to south and no species further than a few degrees west to east
except for L. oblongifolia. The majority of Andean species occur
at relatively low altitudes, mainly ⬍1,500 m (31). The wide range
of Ladenbergia outside the Andean area is entirely attributable
to a small cluster of species, which appeared to be closely related
with each other according to morphology (31). The only exceptions are two species that are not related to these: Ladenbergia
oblongifolia, which has its main range in the Andes, and also has
a disjunct occurrence in the Serra da Neblina, and L. hexandra,
which is endemic to the mountains of southeastern Brazil.
The distribution range of Cinchona (Fig. S2 A) is Andean,
except for C. pubescens, which extends its range into southern
Central America and the coastal mountains of Venezuela. Like
Ladenbergia, Cinchona has a predominantly premontane to
montane range. All records outside this range are probably
introductions and, occasionally, subsequent naturalization (32).
The distribution range of Ciliosemina (Fig. S2B) falls largely
within the Northern Andes, but reaches marginal parts of the
Amazon basin.
Maguireocharis neblinae is a montane species growing at
considerable altitude in the Serra da Neblina (Fig. S2A). In the
absence of sequence data, and because of its unrevealing morphology, its relationships are obscure (1).
The monotypic genus Pimentelia (Fig. S2B) occurs on the
eastern slopes of the Andes in southern Peru and northern
Bolivia, whereas Stilpnophyllum (Fig. S2D) occurs in the Eastern
Cordillera of northern Peru and southern Ecuador, and in the
Central Cordillera of northern Colombia. Pimentelia could not
be sequenced, but morphology indicates that it is the sister group
of Stilpnophyllum (1). These 6 related species are all confined to
the Andes. Stilpnophyllum appears to have a wide distribution
gap between southern Ecuador and Colombia. This is likely to
be real, because montane forests of eastern Ecuador are fairly
well explored. Montane forests of northern and central Peru,
however, are poorly explored, so the number of localities shown
in this region is probably an underestimate.
The range of Joosia (Fig. S2C) extends over most of the
tropical Andean chain, and it is almost fully confined to it. One
species, J. umbellifera, extends throughout the range of the genus
and is the only species occurring north of 1°N. Except for the
single locality of J. multiflora in southernmost Peru, J. umbellifera
is also the only species occurring south of 11°S. The largest
species diversity of Joosia occurs in Ecuador and northern Peru,
although species richness is not great anywhere. The slight drop
seen in northernmost Peru is almost certainly an artifact caused
by poor exploration of this region. Most species occur in the
premontane to lower montane life zones, but 3 have only been
found in the lowlands, and only 1 in the montane zone (30).
The distribution range of Cinchonopsis amazonica (Fig. S1E),
the only species of the genus, comprises most of the western half
of the Amazon Basin, and adjacent parts of the Orinoco drainage. Scattered information on herbarium labels suggests that it
grows mainly in upland forest on white sand, which also agrees
with the single field observation made by one of us (A.A.).
3 of 20
The range of Remijia (Fig. S1D) comprises mainly the central
parts of the Amazon basin (between ⬇55°W and 74°W) and
adjacent parts of the Guiana Shield. In addition, there are 2
apparently disjunct areas, 1 in the western Amazonia and 1 in the
Serra do Espinhaço in southeastern Brazil. Each of these
exclaves has but a single endemic species (R. chelomaphylla and
R. ferruginea, respectively). Remijia has a very pronounced center
of diversity in central Amazonia, mainly in the Rio Negro, Rio
Branco, and Rio Madeira basins, but extending into the Guiana
Highlands. This seems to be related to an ecological preference
of Remijia species for savanna and Amazonian caatinga, biomes
that are particularly common in these regions (e.g., ref. 67).
However, this is not the only explanation, because some central
Amazonian species seem to occur in the rain forest. The
phylogeny of Andersson and Antonelli (1) showed 2 sister
groups, R. chelomaphylla–R. macrocnemia and R. pacimonica–R.
ulei. The first 2 species occur only west of the diversity center, R.
chelomaphylla in the western exclave and R. macrocnemia in a
relatively small range west and northwest of the Trapecio
Amazónico, where it is the only species. The other 2 species
occur within the region of high species richness and are morphologically very similar. The apparent east–west sister group
relationship may be an artifact, however, because many Remijia
species have not yet been sampled.
Effects of Taxon Sampling on Biogeographic Reconstruction and Age
Estimates. Biogeographic reconstruction. Barber and Bellwood (68)
showed that biogeographical optimization methods such as
dispersal-vicariance analysis could be highly sensitive to incomplete taxon sampling, especially if the missing taxa occupy a basal
position in the phylogeny and/or occur in areas that are underrepresented in the analysis.
Although taxon sampling may be a problem in a large family
such as Rubiaceae, spanning nearly all continents, we have tried
to overcome this problem by using very encompassing areas
outside the Neotropics, where most of the Rubioideae and
Ixoroideae are found, as well as the first diverging clade in
Cinchonoideae, the tribe Naucleeae. Although the small tribes
Hamelieae, Hillieae, and Chiococceae are poorly represented in
number of taxa, the distribution of the species included here are
representative of their tribes, which are all predominantly distributed in Central America and the Caribbean region. A
possible source of ‘‘noise’’ in the optimizations may be caused by
Guettarda speciosa, an Australasian species that alone represents
the tribe Guettardeae. Except for that species, the tribe has a
Neotropical distribution (mainly in Central America and the
West Indies; ref. 69). To estimate the effect of this bias in
ancestral area optimizations, we carried out a new analysis
coding the Guettarda lineage as restricted to Central America,
the West Indies, and both. Except for the ancestor of the clade
comprising Guettarda and Rogiera, the new coding did not affect
any other ancestral area optimizations.
For tribes Cinchoneae and Isertieae, which are the focus of
this article and upon which our main biogeographic conclusions
depend, the key areas are the Northern and Central Andes (areas
C and D in Fig. 1 Inset). In special, the presence of Central
Andean species in a basal position within these tribes could
potentially alter the conclusions drawn based on the empirical
sequences analyzed.
To test the sensitivity of DIVA reconstructions to incomplete
taxon sampling, we first estimated the geographic bias in our
sample for each genus included in tribes Isertieae and Cinchoneae. We compiled distribution data for all species, genera, and
tribes (Table S4) and compared the representation of each area
in the complete inventory against that of our molecular sample.
Fig. S6 (left column) shows that the geographical distribution of
species in our sample is fairly representative of the actual
distribution of the specie. For tribes, area C is slightly underAntonelli et al. www.pnas.org/cgi/content/short/0811421106
represented in Cinchoneae and area D in Isertieae. In contrast,
area F (Amazonia) is fairly represented in both tribes. For
genera, the geographical bias is largest in Joosia, Stilpnophyllum,
and Cinchona, underrepresented in area D, and Remijia, which
is missing all 8 species endemic to the Guiana Shield and the
Brazilian Planalto (areas G and H). Some of these genera are
also poorly represented in botanical collections: Most species of
Stilpnophyllum, and many of Joosia, are known from the type
specimen only (Fig. S6, right column). The paucity of specimens
in botanical collections directly reflects the difficulty to find
these species in the field, as evidenced by numerous botanical
expeditions undertaken from our research institute in the last
decades, as well as by the experience of many colleagues working
locally. Although this paucity may sometimes be attributed to
poor botanical exploration of some inaccessible regions, most
species seem to be naturally rare. In addition, many species are
only minimally distinct, which calls into question the species
concept applied within each genus and suggests that some of the
rarest species could in fact be variations of a more common
species, or hybrids (e.g., ref. 32).
Based on this survey (Fig. S6), we then followed a similar
approach to Barber and Bellwood (68). We performed a series
of heuristic analyses in which a hypothetical missing taxon was
added, one at a time, at certain points in the phylogeny. Instead
of random addition, we kept the basic backbone of the phylogeny
(all major nodes for Cinchonoideae genera) as we did for the
temporal simulations (see below). The phylogenetic position of
the missing taxa within each genus was chosen following morphological cladograms whenever available (e.g., for genera
Joosia, Ladenbergia). When no morphological phylogeny was
available (e.g., genus Remijia) or did not include any additional
species (genus Isertia), the missing taxon was added, alternatively, to the most basal or the most distal node within the genus.
The distribution of the missing (hypothetical) taxon was determined following the geographical bias estimated for each genus
(see Fig. S6).
Fig. S7 shows the results of these simulations. Somewhat
surprisingly, DIVA ancestral area reconstructions proved to be
very stable to the addition of missing taxa. Adding hypothetical
taxa from underrepresented areas to the phylogeny did not alter
the original ancestral area reconstructions (with no taxon addition) for the key nodes in the Isertieae/Cinchoneae phylogeny.
The only exception was node 51: The addition of a taxon from
areas G or H to the base of the Remijia clade (J6, Fig. S7A )
resulted in the ancestor of Remijia, Ciliosemina, and Ladenbergia
being originally distributed in areas G and/or H (J6, Fig. S7B).
However, this has no consequences for our biogeographic scenario, because ancestral area reconstructions for the rest of the
nodes remained unaltered (Fig. S7B). That is, Remijia would
have reached the Guiana Shield and Southeastern South America (areas G and H, respectively) earlier than we postulate in our
scenario, but biogeographical inferences concerning the Western Andean Portal and Lakes Pebas would not change. Moreover, these changes in the ancestral distribution of Remijia would
only occur if the missing taxon really is sister to the rest of the
genus: A distal position among the remaining ⬇40 species would
instead produce the same outcome as in our original reconstruction (J7, Fig. S7).
The only scenario that could significantly alter our hypotheses
is if the 2 species from area D (Central Andes) in genus Joosia
(J. multiflora and J. dichotoma) and the 2 species from area D in
genus Isertia (I. krausei and I. reticulata) came to occupy the 2
most basal positions within the phylogeny of each of these genera
(simulations not shown). This would result in area D being
inferred as the ancestral area of Joosia and of Isertia, and area
CD as the ancestral area of tribe Cinchoneae. In other words,
tribe Cinchoneae would have been originally present in the
North and Central Andes instead of dispersing to the Central
4 of 20
Andes from the north after the uplift of the WAP as suggested
by our original reconstruction. However, the probability that the
2 Central Andean species of Joosia would occupy the 2 most
basal positions in the phylogeny of the genus is low. In Andersson’s (30) morphological phylogeny, these 2 species are not
phylogenetically close: Joosia multiflora occupies the most basal
position in the cladogram, whereas J. dichotoma is ‘‘firmly nested
within the genus’’ (p. 28 in ref. 30). Both Joosia multiflora and J.
dichotoma are known from the type collection only, and we have
been unable to find them in the field for sequencing. In the case
of Isertia, Andersson’s (70) morphological phylogeny of Isertieae
did not include any Central Andean species. In the only published revision of the genus, Boom (29) included Isertia krausei
and I. reticulata within section Cassupa, which is separated from
section Isertia on the basis of the presence of fruits with fleshy
endocarp. However, our molecular phylogeny does not support
Boom’s infrageneric classification: Isertia pittieri and I. laevis
from section Cassupa appear nested within a clade composed
exclusively by species from section Isertia. Moreover, Isertia
krausei is known from the type collection only, whereas I.
reticulata is ‘‘a seldom collected and imperfectly known species’’
that may occur also in Colombia (29).
Out of the eight species of Cinchona endemic to the Central
Andean region, only one (C. calisaya) was possible to include in
our analysis. Our molecular phylogeny places Cinchona calisaya
as the sister group to the rest of the genus, and DIVA infers the
Northern and Central Andes (area CD) as the geographic origin
of Cinchona (node 46). Even if the other 7 Central Andean
species in the genus come out as a basal grade in the phylogeny—an unlikely inference according to Andersson’s (32) morphological cladogram, which places all Central Andean species
in different clades—and D is inferred as the ancestral area of
Cinchona (with later dispersal to C), this would not affect the
ancestral reconstructions for the backbone nodes in Cinchoneae
(nodes 44, 45, and 51), which still is inferred as having originated
in the Northern Andes, area C.
All things considered, we believe that the possibility of missing
taxa from area D to have a significant influence in our scenario
is low and that the DIVA results presented here are stable to
incomplete taxon sampling.
Age estimates. Linder et al. (71) demonstrated that limited taxon
sampling may influence age estimates, generally resulting in
younger ages for nodes than those obtained with a denser
sample. To investigate these effects in the Rubiaceae, we followed a similar approach to Linder et al. (71): first, we identified
14 ‘‘core’’ nodes across the phylogeny corresponding to all
genera in tribes Cinchoneae and Isertieae, main clades in
Cinchonoideae including the Cephalanthus calibration point,
subfamilies Rubioideae and Ixoroideae, and outgroup (nodes 2,
7, 13, 22, 25, 27, 32, 33, 41, 43, 46, 52, 55, and 56 in Fig. S4);
second, we generated reduced matrices comprising 30%, 40%,
60%, and 80% of the original dataset by randomly deleting
terminal taxa, while still keeping at least 2 species from each core
node in order to keep the backbone of relationships in the
phylogeny (the basic structure of the phylogeny); third, we
perfomed Bayesian phylogenetic analyses and computed meanlength consensus phylograms for each of the reduced matrices,
using the same settings as for the original dataset; and last, we
estimated divergence times for each of those phylograms using
the same methodology as previouly described.
The results of these analyses are summarized in Fig. S8.
Although there is a slight tendency to underestimation of ages
when taxon sampling decreases (Fig. S8A), for all core nodes in
the phylogeny the ages obtained for the reduced matrices fall
within the 95% confidence intervals estimated from complete
dataset (Fig. S8B). Similarly, even a 2-fold increase in the
number of taxa (trend lines in Fig. S8A) is not expected to result
in node ages outside the confidence intervals of particular nodes.
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
The only exceptions are nodes 20, 31, and 33 in the dataset
comprising only 30% of the original taxa: their ages are underestimated by 1.3, 0.7, and 0.53 Ma from the original confidence
intervals, respectively. It seems that 30% represents a threshold
level for those nodes, after which ages stabilize.
These results indicate that the addition of more taxa in
Rubiaceae is not expected to significantly influence the empirical results presented, nor their temporal interpretation. Moreover, they support the finding by Linder et al. (71) that Penalized
Likelihood is largely insensitive to taxon sampling, as compared
with other dating methods such as nonparametric rate smoothing
and Bayesian dating.
Generality of the Biogeographic Patterns Found. Western Andean
Portal (WAP). Although it has never been properly analyzed, it is
particularly noteworthy that several studies have demonstrated
a biogeographic disjunction between the Northern and Central
Andes, roughly corresponding to the latitude used in this study
for separating these 2 operational areas (⬇5°S). However, the
dispersal barrier between these regions has received many
different denominations in the literature, which seem to refer to
approximately the same area: Western Portal, Andean Gate,
Marañón Portal, Guayaquil Gap, Huancabamba Depression,
Huancabamba Deflexion, Northern Peruvian Low, and Pirua
Divide.
In plants, the pattern has been demonstrated in many families,
such as Campanulaceae (72), Calceolariaceae (73), Tropaeolaceae (74), Loasaceae, Passifloraceae, Grossulariaceae (54, 75),
and Alstroemeriaceae (76). In animals, Vuilleumier (77) showed
such a disjunction for populations of the bird superspecies
Asthenes flammulata (Furnariidae), Nores (78) defined the region north of this barrier (northwestern Andes) as a general area
of bird endemism, and Cortés-Ortiz (79) defined this barrier as
the southern limit of the monkey Alouatta palliata. Moreover,
this region (the Rı́o Marañon in Peru) has long been recognized
as the turnover point between the Northern and Central Andean
regions of bird endemism (80), further emphasizing the role of
the WAP as a major biogeographic barrier. We have not
attempted to compile an exhaustive account of taxa whose
distribution is affected by the barrier, but anticipate that many
other groups may show similar patterns.
Although our results demonstrate how and when the WAP
influenced the distribution of a modern group of plants, they
cannot provide evidence to its exact nature, geographic extension, and duration. The few paleontological studies in the area
strongly support the existence a marine incursion that reached
the western part of present-day Amazonia at least during the
Late Eocene (81, 82) and Middle Miocene (53), but how long
marine settings dominated the region before and after these
epochs is a matter of debate. Moreover, contrary to several
earlier studies (e.g., ref. 83), new marine geophysical data suggest
that global sea levels may have been relatively stable from the
Early Eocene onwards (ref. 84 and references therein). Therefore it seems plausible that a marine incursion dominated the
WAP during most of the Eocene–Middle Miocene, ending with
the uplift of the Eastern Cordillera.
The geographic extent of the WAP has also been under
debate. From the Pacific coast, marine settings have been
demonstrated to reach as far east as ⬇75° W in the Late Eocene
(82) and ⬇79° W in the Middle Miocene (53). An aquatic
connection between the Pacific and Amazonia has long been
hypothesized (e.g., 53, 85, 86), but no conclusive evidence seems
yet to support such a long connection (F. P. Wesselingh and
C. Hoorn, personal communication). However, there is some
evidence that intermittent connections existed between Lake
Pebas in western Amazonia and the Pacific through the Ecuadorean Andes during the Middle Miocene (87). Even if the WAP
would not have been filled by sea water during its entire existence
5 of 20
and even if marine incursions did not reach as far east as
Amazonia, it could still have acted as a dispersal barrier to
montane organisms: either by being lowland corridors or by
imposing ecological constraints (e.g., very dry climates were
present in southern Ecuadorean Andes in the Miocene; ref. 87).
We make here a plea for more studies that address these
important issues.
Lake Pebas. The biogeographic scenario proposed here implies
that all plant species adapted to dry land conditions (terra firme)
in present-day western Amazonia have gained their current
distribution after Lake Pebas drained (i.e., in the last ⬇11 Ma;
ref. 88). Similarly, in situ speciation in those terrestrial groups
could not have taken place before that event.
These results may seem controversial given the fact that
western Amazonia is characterized today by an outstanding
number of endemic plant species, of which many belong to
long-lived woody families not expected to speciate fast. In a
recent study using large molecular chronograms from 5 angio-
sperm families, Smith and Donoghue (89) showed that there is
a strong correlation between growth mode and speciation rates:
Herbs generally diversify much more rapidly than shrubs and
trees. However, the few molecular dating studies that have dealt
with a representative number of Amazonian trees have presented very contrasting results. Two of the largest genera of
Neotropical trees have been inferred to have diversified recently:
Inga (Fabaceae, 300 spp), which is estimated to have started its
diversification either 9.8 Ma (based on cpDNA) or 1.6 Ma (based
on nrDNA; ref. 90); and Guatteria (Annonaceae, 265 spp), where
the crown age for all Neotropical taxa (including 2 large Amazonian subclades) is estimated at 7.4 ⫾ 1.4 Ma (91). Among
animals, a recent study (92) suggested that the diversification of
Amazonian poison frogs (Dendrobatidae) also occurred recently, with radiations in the last 10 Ma. These results fit well into
the paleogeographic scenario for Amazonia proposed here and
suggest that Amazonian organisms may have evolved at an
unusually fast rate after the end of Lake Pebas.
1. Andersson L, Antonelli A (2005) Phylogeny of the tribe Cinchoneae (Rubiaceae), its
position in Cinchonoideae, and description of a new genus, Ciliosemina. Taxon 54:17–
28.
2. Albach DC, Soltis PS, Soltis DE, Olmstead RG (2001) Phylogenetic analysis of asterids
based on sequences of four genes. Ann Mo Bot Gard 88:163–212.
3. Katoh K, Kuma K-i, Toh H, Miyata T (2005) MAFFT version 5: Improvement in accuracy
of multiple sequence alignment. Nucleic Acids Res 33:511–518.
4. Antonelli A (2008) Higher level phylogeny and evolutionary trends in Campanulaceae
subfam. Lobelioideae: Molecular signal overshadows morphology. Mol Phylogen Evol
46:1–18.
5. Farris JS, Albert VA, Källersjö M, Lipscomb D, Kluge AG (1996) Parsimony jackknifing
outperforms neighbor-joining. Cladistics 12:99 –124.
6. Swofford DL (2002) PAUP*. Phylogenetic Analysis Using Parsimony (*and other
methods) (Sinauer, Sunderland, MA), Ver 4.0b10.
7. Pol D (2004) Empirical problems of the hierarchical likelihood ratio test for model
selection. Syst Biol 53:949 –962.
8. Posada D, Buckley T (2004) Model selection and model averaging in phylogenetics:
Advantages of Akaike information criterion and Bayesian approaches over likelihood
ratio tests. Syst Biol 53:793– 808.
9. Nylander JAA (2004) MrModeltest (Evolutionary Biology Centre, Uppsala Univ, Uppsala, Sweden), Version 2.
10. Ronquist F, Huelsenbeck JP (2003) MrBayes3: Bayesian phylogenetic inference under
mixed models. Bioinformatics 19:1572–1574.
11. Ronquist F, Huelsenbeck JP, van der Mark P (2006) MrBayes 3.1 Manual. Available
online at http://mrbayes.csit.fsu.edu/manual.php (Florida State Univ, Talahassee, FL).
12. Nylander JAA, Wilgenbusch JC, Warren DL, Swofford DL (2008) AWTY: Are we there
yet? A system for graphical exploration of MCMC convergence in Bayesian phylogenetics. Bioinformatics 24:581–583. Available online at http://ceb.scs.fsu.edu/awty.
13. Felsenstein J (1981) Evolutionary trees from DNA sequences: A maximum likelihood
approach. J Mol Evol 17:368 –376.
14. Sanderson MJ (2002) Estimating absolute rates of molecular evolution and divergence
times: A penalized likelihood approach. Mol Biol Evol 19:101–109.
15. Sanderson MJ (2002b) r8s (Univ of California, Davis, CA) Version 1.70,
http://loco.biosci.arizona.edu/r8s/.
16. Bremer K, Friis EM, Bremer B (2004) Molecular phylogenetic dating of asterid flowering
plants shows early Cretaceous diversification. Syst Bot 53:496 –505.
17. Collinson ME, Boulter MC, Holmes PL (1993) Magnoliophyta (‘Angiospermae’). The
Fossil Record 2, ed Benton MJ (Chapman & Hall, London).
18. Mai DH, Walther H (1985) The upper Eocene floras of the Weisselster basin and
adjacent areas. (in German) Abh Staat Mus Miner Geol Dresden 33:1–260.
19. Raniecka-Bobrowska J (1959) The Tertiary spermatophyte flora from Konin. (in Polish)
Biul Inst Geol 130:159 –252.
20. Dorofeev PI (1960) New data about Tertiary floras of Kirrenskiy ravine on the Ob River.
(in Russian) Dokl Akad Nauk SSSR 133:211–213.
21. Dorofeev PI (1963) Tertiary Floras of Western Siberia. (in Russian) (Izd Akad Nauk SSSR,
Moscow).
22. Friis EM (1985) Angiosperm fruits and seeds from the Middle Miocene of Jutland,
Denmark. Biol Skr 24:1–165.
23. Gradstein FM, et al. (2004) A Geologic Time Scale 2004 (Cambridge Univ Press,
Cambridge, UK).
24. Berry EW (1938) Tertiary flora from the Rio Pichileufu, Argentina. Geol Soc Am Spec
Paper 12:1–149.
25. Berry EW (1929) Early Tertiary fruits and seeds from Belén Peru. Johns Hopkins Univ
Studies Geol 10:139 –172.
26. Nepokroeff M, Bremer B, Sytsma KJ (1999) Reorganization of the genus Psychotria and
tribe Psychotrieae (Rubiaceae) inferred from ITS and rbcL sequence data. Syst Bot
24:5–27.
27. Hodkinson TR, Parnell JAN (2006) in Introduction to the Systematics of Species Rich
Groups. Reconstructing the Tree of Life: Taxonomy and Systematics of Species Rich
Taxa (CRC, Boca Raton, FL), pp 3–20.
28. Drummond AJ, Rambaut A (2007) BEAST: Bayesian evolutionary analysis by sampling
trees. BMC Evol Biol 7:214.
29. Boom BM (1985) A revision of Isertia (Isertieae: Rubiaceae). Brittonia 36:425– 454.
30. Andersson L (1997a) A new revision of Joosia (Rubiaceae–Cinchoneae). Brittonia
49:24 – 44.
31. Andersson L (1997b) Synopsis of the genus Ladenbergia (Rubiaceae). Nord J Bot
17:255–299.
32. Andersson L (1998) A revision of the genus Cinchona (Rubiaceae–Cinchoneae). Mem
NY Bot Gard 80:1–75.
33. Steyermark J (1972) Rubiaceae. The Botany of the Guayana Highland. Mem NY Bot
Gard 23:227– 832.
34. Steyermark J (1974) Rubiaceae in Flora de Venezuela, eds Lasser T, Steyermark JA
(Instituto Botánico, Caracas), Vol 9, pp 1–3.
35. Andersson L (1994) Tribe 1. Cinchoneae in Flora of Ecuador, eds Harling G, Andersson
L (Univ of Gothenburg, Gothenburg, Sweden), Vol 50, pp 3– 82.
36. Andersson L (1999) Isertia in Flora of Ecuador, eds Harling G, Andersson L (Univ of
Gothenburg, Gothenburg, Sweden), Vol 62, pp57– 65.
37. Elias TS (1976) A Monograph of the Genus Hamelia (Rubiaceae). Mem NY Bot Gard
26(4):81–144.
38. Kirkbride JH, Jr (1985) Manipulus rubiacearum IV. Kerianthera (Rubiaceae), a new
genus from Amazonian Brazil. Brittonia 37:109 –116.
39. Taylor CM, et al. (2004) Poaceae–Rubiaceae in Flora of the Venezuelan Guayana, eds
Steyermark JA, et al. (Mo Bot Gard Press, St. Louis), Vol. 8.
40. Andersson L (1992) A provisional checklist of Neotropical Rubiaceae. Scripta Bot Belg
1:1–199.
41. Bridson D, Verdcourt B (1988) Rubiaceae (Part 2) in Flora of Tropical East Africa, ed
Polhill RM (Rotterdam, AA Balkema), pp 415–747.
42. Mabberley DJ (2002) The Plant Book (Cambridge Univ Press, Cambridge, UK).
43. Puff C (1991) The genus Paederia L.—A multidisciplinary study. Opera Bot Belg
3:6 –375.
44. Smith AC (1988) Rubiaceae. Flora Vitiensis Nova 4:143–376.
45. Taylor DW (1991) Paleobiogeographic relationships of Andean angiosperms of Cretaceous to Pliocene age. Palaeogeogr Palaeoclimatol 88:69 – 84.
46. Gregory-Wodzicki KM (2000) Uplift history of the central and northern Andes: A
review. Geol Soc Am Bull 112:1091–1105.
47. Cracraft J (1988) Deep-history biogeography, retrieving the historical pattern of
evolving continental biotas. Syst Zool 37:221–236.
48. Morrone JJ (1994) Distributional patterns of species of Rhytirrinini (Coleoptera:Curculionidae) and the historical relationships of the Andean provinces. Glob Ecol Biogeogr Lett 4:188 –194.
49. Posadas PE, Estévez JM, Morrone JJ (1997) Distributional patterns and endemism areas
of vascular plants in the Andean subregion. Fontqueria 48:1–9.
50. Katina L, Morrone JJ, Crisci JV (1999) Track analysis reveals the composite nature of the
Andean biota. Aust J Bot 47:111–130.
51. Briggs JC (1994) The genesis of Central America: Biology versus geophysics. Glob Ecol
Biogeogr 4:169 –172.
52. Hoorn C (1993) Marine incursions and the influence of Andean tectonics on the
Miocene depositional history of northwestern Amazonia: Results of a palynostratigraphic study. Palaeogeogr Palaeoclimatol 105:267–309.
53. Hungerbühler D, et al. (2002) Neogene stratigraphy and Andean geodynamics of
southern Ecuador. Earth Sci Rev 57:75–124.
54. Weigend M (2004) Additional observations on the biogeography of the AmotapeHuancabamba zone in Northern Peru: Defining the South-Eastern limits. Rev Peru Biol
11:127–134.
55. Brumfield RT, Capparella AP (1996) Historical diversification of birds in Northwestern
South America: A molecular perspective on the role of vicariant events. Evolution
(Lawrence, Kans) 50:1607–1624.
56. Scotese CR (2001) Atlas of Earth History. (Paleomap Project, Arlington, TX). Vol 1,
Paleogeography.
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
6 of 20
57. Morley RJ (2003) Interplate dispersal routes for megathermal angiosperms. Perspec Pl
Ecol Evol Syst 6:5–20.
58. Sanmartı́n I (2005) A paleogeographic history of the Southern Hemisphere (Uppsala
Univ, Uppsala, Sweden) (www.ebc.uu.se).
59. Ronquist F (1996) DIVA (Uppsala Univ, Uppsala, Sweden), Version 1.1 (www.ebc.uu.se/
systzoo/research/diva/diva.html).
60. Ronquist F (1997) Dispersal-vicariance analysis: A new approach to the quantification
of historical biogeography. Syst Biol 46:195–203.
61. Sanmartı́n I, Ronquist F (2004) Southern Hemisphere biogeography inferred by eventbased models: plant versus animal patterns. Syst Biol 53:216 –243.
62. Ree RH, Moore BR, Webb CO, Donoghue MJ (2005) A likelihood framework for
inferring the evolution of geographic range on phylogenetic trees. Evolution (Lawrence, Kans) 59:2299 –2231.
63. Nylander JAA, Olsson U, Alström P, Sanmartı́n I (2008) Accounting for phylogenetic
uncertainty in biogeography: A Bayesian approach to dispersal-vicariance analysis of
the Thrushes (Aves: Turdus). Syst Biol 57:257–268.
64. Maddison WP, Maddison DR (2007) Mesquite: A modular system for evolutionary
analysis (Mesquite Software, Austin, TX), Version 2.01 (mesquiteproject.org).
65. Manen JF, Dessein S, De Block P, Robbrecht E (2006) Towards an evolutionary scenario
for the coffee family (Rubiaceae, angiosperms): Further insights from a supertree.
Third International Rubiaceae Conference, published abstract. Scripta Bot Belg 40:47.
66. Sónke B, Dessein S, Taedoumg H, Groeninckx I, Robbrecht E (2008) A new species of
Colletoecema (Rubiaceae) from southern Cameroon with a discussion of relationships
among basal Rubioideae. Blumea 53:533–547.
67. Hueck K, Seibert P (1972) Vegetation Map of South America (Gustav Fischer, Stuttgart,
Germany) (in German).
68. Barber PH, Bellwood DR (2005) Biodiversity hotspots: Evolutionary origins of biodiversity in wrasses (Halichoeres: Labridae) in the Indo-Pacific and new world tropics. Mol
Phylogen Evol 35:235–253.
69. Achille F, Motley TJ, Lowry PP, II, Jérémie J (2006) Polyphyly in Guettarda L. (Rubiaceae,
Guettardeae) based on nrDNA ITS sequence data. Ann Mo Bot Gard 93:103–121.
70. Andersson L (1996) Circumscription of the tribe Isertieae (Rubiaceae). Opera Bot Belg
7:139 –164.
71. Linder HP, Hardy CR, Rutschmann F (2005) Taxon sampling effects in molecular clock
dating: An example from the African Restionaceae. Mol Phylogen Evol 35:569 –582.
72. Ayers T (2000) Biogeography of Lysipomia: An illustration of species richness adjacent
to the Huancabamba Depression. Arnoldoa 6:13–27.
73. Molau U (1988) Scrophulariaceae I. Calceolarieae. Fl Neotropica 47:1–326.
74. Sparre B, Andersson L (1991) A taxonomic revision of the Tropaeolaceae. Opera Bot
108:1–139.
75. Weigend M (2002) Observations on the Biogeography of the Amotape-Huancabamba
Zone in Northern Peru. Bot Review 68(1):38 –54.
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
76. Hofreiter A, Rodrı́guez R (2006) The Alstroemeriaceae in Peru and neighbouring areas.
Rev Peru Biol 13(1):5– 69.
77. Vuilleumier F (1968) Population structure of the Asthenes flammulata superspecies
(Aves: Furnariidae). Breviora 297:1–21.
78. Nores M (2004) The implications of late Tertiary and Quaternary sea level events for
avian distribution patterns in the lowlands of northern South America. Glob Ecol
Biogeogr 13:149 –161.
79. Cortés-Ortiz L, et al. (2003) Molecular systematics of the Neotropical monkey genus
Alouatta. Mol Phylogen Evol 26:64 – 81.
80. Stotz DF, Fitzpatrick JW, Parker TA, Moskovits DK (1996) Neotropical Birds: Ecology and
Conservation (Univ of Chicago Press, Chicago).
81. Lundberg JG, et al. (1998) in Phylogeny and Classification of Neotropical Fishes, eds
Malabarba LR, Reis RE, Vari RP, Lucena ZM, Lucena CAS (Edipucrs, Porto Alegre, Brazil),
pp 13– 48.
82. Santos C, Jaramillo C, Bayona G, Rueda M, Torres V (2008) Late Eocene marine incursion
in north-western South America. Palaeogeogr Palaeoclimatol 264:140 –146.
83. Haq BU, Hardenbol J, Vail PR (1987) Chronology of fluctuating sea levels since the
Triassic. Science 235:1156 –1167.
84. Müller RD, Sdrolias M, Gaina C, Steinberger B, Heine C (2008) Long-term sea-level
fluctuations driven by ocean basin dynamics. Science 319:1357–1362.
85. Katzer F (1903) Grundzüge der Geologie des Unteren Amazonasgebietes. (Max Weg,
Leipzig, Germany).
86. Nuttal CP (1990) A review of the Tertiary non-marine molluscan faunas of the Pebasian
and other inland basins of north-western South America. Bull Brit Mus Nat Hist Geol
45:165–371.
87. Steinmann M, Hungerbühler D, Seward D, Winkler W (1999) Neogene tectonic evolution and exhumation of the southern Ecuadorian Andes; a combined stratigraphy and
fission-track approach. Tectonophysics 307:255–276.
88. Wesselingh FP, et al. (2010) in Amazonia: Landscape and species evolution, eds
Wesselingh FP, Hoorn C (Blackwell Publishing, Oxford), in press.
89. Smith SA, Donoghue MJ (2008) Rates of molecular evolution are linked to life history
in flowering plants. Science 322:86 – 89.
90. Richardson JE, Pennington RT, Pennington TD, Hollingsworth PM (2001) Rapid diversification of a species-rich genus of Neotropical rain forest trees. Science 293:2242–
2245.
91. Erkens RHJ, Chatrou LW, Maas JW, van der Niet T, Savolainen V (2007) A rapid
diversification of rainforest trees (Guatteria; Annonaceae) following dispersal from
Central into South America. Mol Phylogen Evol 44:399 – 411.
92. Santos JC, et al. (2009) Amazonian amphibian diversity is primarily derived from Late
Miocene Andean lineages. PLoS Biol 7:e56.
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Fig. S1. Distribution patterns in Neotropial Rubiaceae: tribes Isertieae and Cinchoneae. (A) Geographical range and species richness of Rubiaceae tribe Isertieae
s. str. (based on maps in ref. 29, specimen citations in ref. 38, and localities cited by ref. 36). (B) Geographical range and species richness of Rubiaceae tribe
Cinchoneae. (C) Geographical range and species richness of the genus Ladenbergia (based on data from ref. 31). (D) Geographical range and species richness
of the genus Remijia (based on Lennart Andersson’s unpublished notes). (E) Recorded localities of Cinchonopsis amazonica (based on Lennart Andersson’s
unpublished notes). (F) Branch tip with inflorescences of Ciliosemina pedunculata and detail of a dissected flower (branch based on herbarium specimen Lleras
et al. in Prance 16958, S and flower detail on St. John and Arcila 20627, GB. [Flower image in F reproduced with permission from ref. 1 (Copyright 2005, Olof Helje).
Branch image in F reproduced with permission from ref. 35 (Copyright 1994, University of Gothenburg).]
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
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Fig. S2. Distribution patterns in tribe Cinchoneae. (A) Geographical range and species richness of the genus Cinchona (based on data from ref. 32), and localities
of Maguireocharis neblinae recorded by ref. 33. (B) Recorded localities of Ciliosemina and Pimentelia (based on Lennart Andersson’s unpublished notes). (C)
Recorded occurrences of the genus Joosia (based on data from ref. 30). (D) Geographical range and species richness of the genus Stilpnophyllum (based on
specimen citations in ref. 35, and Lennart Andersson’s unpublished notes).
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
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.97
89
.98
69 1.00
100
.59
54 1.00
100 1.00
99 .91
98 1.00
100 1.00
100
.65
1.00
97
1.00
.97
100
79
1.00
77 1.00
100 1.00
100
1.00
100
1.00
100
1.00
100 1.00
100 1.00
99
1.00
100
1.00
100
.87
72 1.00
100 1.00
100
1.00
100
.74
83
1.00
100
.35
1.00
100
1.00
100
.41
1.00
98
1.00
100
1.00
100
1.00
84
.79
56 1.00
99
.36
1.00
100
1.00
100
1.00
100
1.00
100 1.00
100 1.00
91 .97
76 .91
.95
67
73
.98
1.00 86
100 1.00
.40
100
.71
1.00
99 1.00
1.00
63
1.00
59 1.00
95
Gelsemium sempervirens
Thevetia peruviana
Geniostoma rupestre
Strychnos nux-vomica
Luculia grandiflora
Ophiorrhiza mungos
Pauridiantha sp.
Lasianthus batangensis
Psychotria kirkii
Paederia foetida
Phyllis nobla
Condaminea corymbosa
Posoqueria latifolia
Sipanea biflora
Mussaenda raiateensis
Sabicea aspera
Vangueria madagascariensis
Genipa americana
Mitriostigma axillare
Hymenodictyon floribundum
Cephalanthus occidentalis
Hallea rubrostipulata
Nauclea orientalis
Guettarda speciosa
Rovaeanthus suffrutescens
Cosmibuena grandiflora
Hamelia papillosa
Cubanola domingensis
Chiococca alba
Exostema lineatum
Kerianthera preclara
Isertia coccinea
Isertia hypoleuca
Isertia haenkeana
Isertia laevis
Isertia pittieri
Isertia rosea
Isertia parviflora
Isertia spiciformis
Joosia aequatoria
Joosia umbellifera
Stilpnophyllum grandifolium
Stilpnophyllum oellgaardii
Cinchonopsis amazonica
Cinchona calisaya
Cinchona pitayensis
Cinchona macrocalyx
Cinchona officinalis
Cinchona mutisii
Cinchona pubescens
Remijia chelomaphylla
Remijia macrocnemia
Remijia pacimonica
Remijia ulei
Ciliosemina pedunculata
Ladenbergia amazonensis
Ladenbergia sp.
Ladenbergia macrocarpa
Ladenbergia carua
Ladenbergia oblongifolia
Ladenbergia pavonii
Fig. S3. Summary of the Bayesian phylogenetic analysis (50% majority-rule consensus tree with compatible groups added). A thick-lined branch indicates that
the branch was also present in the majority-rule consensus tree of the parsimony-jackknife analysis. Numbers above branches indicate the posterior probability
of the clade. Numbers below branches show jackknife support values, whenever applicable. Brackets identify subfamilies and tribes (CHI, Chiococceae s. lat.; CIN,
Cinchoneae s. str.; GUE, Guettardeae; HAM, Hamelieae; HIL, Hillieae; ISE, Isertieae s. str.; NAU, Naucleeae s. lat.; RON, Rondeletieae).
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
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Fig. S4. Molecular dating analysis of the Rubiaceae. (A) Chronogram showing 95% confidence intervals of node ages (indicated by bars; n ⫽ 1,000). Tree
topology is the 50% majority-rule Bayesian consensus tree with all compatible groups added. Node numbers refer to Table S2. Red stars indicate calibration points
based on molecular and fossil evidence [Node 1: crown group age of Gentianales as determined in the large-scale analysis of Bremer et al. (16); node 22: fruits
of Cephalanthus kireevskianus from the Late Eocene of Germany reported by Mai and Walther (18)]. (Inset) SEM pictures depicting fossil (I–II) and extant (III–IV)
fruits of Cephalanthus. I, III: Apical part of mericarp with germination valve detached showing characteristic strophiole (ST) and seed (S). II, IV: seed. [Reproduced
with permission from ref. 22 (Copyright 1985, The Royal Danish Academy of Sciences and Letters).] (B) Number of recovered Cephalanthus fruit fossils per geologic
epoch. (C) Locations of all 19 records of Cephalanthus fossilized fruits.
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Lowland (< 500 m)
Montane (> 500 m)
Gelsemium sempervirens
Thevetia peruviana
Geniostoma rupestre
Strychnos nuxvomica
Luculia grandiflora
Ophiorrhiza mungos
Pauridiantha sp
Lasianthus batangensis
Psychotria kirkii
Paederia foetida
Phyllis nobla
Condaminea corymbosa
Posoqueria latifolia
Sipanea biflora
Mussaenda raiateensis
Sabicea aspera
Vangueria madagascariensis
Genipa americana
Mitriostigma axillare
Hymenodictyon floribundum
Cephalanthus occidentalis
Hallea rubrostipulata
Nauclea orientalis
Guettarda speciosa
Rovaeanthus suffrutescens
Cosmibuena grandiflora
Hamelia papillosa
Cubanola domingensis
Chiococca alba
Exostema lineatum
Kerianthera preclara
Isertia coccinea
Isertia hypoleuca
Isertia haenkeana
Isertia laevis
Isertia pittieri
Isertia rosea
Isertia parviflora
Isertia spiciformis
Joosia aequatoria
Joosia umbellifera
Stilpnophyllum grandifolium
Stilpnophyllum oellgaardii
Cinchonopsis amazonica
Cinchona calisaya
Cinchona pitayensis
Cinchona macrocalyx
Cinchona officinalis
Cinchona mutisii
Cinchona pubescens
Remijia chelomaphylla
Remijia macrocnemia
Remijia pacimonica
Remijia ulei
Ciliosemina pedunculata
Ladenbergia amazonensis
Ladenbergia sp
Ladenbergia macrocarpa
Ladenbergia carua
Ladenbergia oblongifolia
Ladenbergia pavonii
Fig. S5. Altitudinal optimization on ancestral nodes. Pie charts represent relative frequencies of optimizations, based on 5,000 Bayesian trees from a stationary
sample. Optimizations performed in the software Mesquite under the Maximum Parsimony (Fitch) criterion, and plotted on the 50% majority-rule consensus
tree (compatible groups added) of Fig. 1. Only trees where the relevant node is present were considered in calculating the relative frequencies.
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
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Geographic distribution of species
Total
Sampled
No. of collections
of each species
Joosia
11
2
23
6
34
6
Cinchona
Ladenbergia
Genus not
revised
Remijia
41
4
4
2
14
8
Tribe
Cinchoneae
118
22
Tribe
Isertieae
15
9
Tribes
Cinchoneae
& Isertieae
133
31
Stilpnophyllum
Isertia
Legends
A
C
1 (= type specimen)
D
E
F
G
H
2–6
>6
Fig. S6. Estimate of geographic bias on taxon sampling for tribes Isertieae and Cinchoneae. (Left) Geographic distribution of all species in the main genera
and tribes of subfamily Cinchonoideae, compared with the distribution of species sampled in our phylogeny. The number of species for each chart is given
immediately after it. Area codings as in Fig. 1. (Right) Number of collected specimens for each species in a genus/tribe. Note that these are minimal numbers,
because only specimens verified in taxonomic revisions have been accounted for (additional specimens may have been collected recently and deposited at various
herbaria).
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A)
32
J 0 33
31
J2
J4
40
42
J1
41 J
3
43 J
5
46
44
45
J6
52
J7
51
55
J8
56
J9
B)
Taxon
addition
(none)
J 0 :D
J 1 :D
J 2 :D
J 3 :D
J 4 :D
J 5 :D
J 6 :G
J 6 :H
J 7 :G
J 7 :H
J 8 :H
J 9 :A
J 9 :H
Kerianthera praeclara
Isertia coccinea
Isertia hypoleuca
Isertia haenkeana
Isertia laevis
Isertia pittieri
Isertia rosea
Isertia parviflora
Isertia spiciformis
Joosia umbellifera
Joosia aequatoria
Stilpnophyllum grandifolium
Stilpnophyllum oellgaardii
Cinchonopsis amazonica
Cinchona calisaya
Cinchona pitayensis
Cinchona macrocalyx
Cinchona officinalis
Cinchona mutisii
Cinchona pubescens
Remijia chelomaphylla
Remijia macrocnemia
Remijia pacimonica
Remijia ulei
Ciliosemina pedunculata
Ladenbergia amazonensis
Ladenbergia sp.
Ladenbergia macrocarpa
Ladenbergia carua
Ladenbergia oblongifolia
Ladenbergia pavonii
Ancestral area reconstructions for key nodes
31
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
32
F
F
F
F
F
F
F
F
F
F
F
F
F
F
33
F
F
F
F
F
F
F
F
F
F
F
F
F
F
40
C
C
C
C
C
C
C
C
C
C
C
C
C
C
41
C
C
C
C
C
C
C
C
C
C
C
C
C
C
42
C
C
C
C
C
C
C
C
C
C
C
C
C
C
43
C
C
C
C
C
C
C
C
C
C
C
C
C
C
44
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
CF
45
C
C
C
C
C
C
C
C
C
C
C
C
C
C
46
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
51
C CF
C CF
C CF
C CF
C CF
C CF
C CF
C CF CG CFG
C CF CH CFH
C CF
C CF
C CF
C CF
C CF
F
F
F
F
F
F
F
F
F
F
F
F
F
F
52
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
CF CDF
55
C
C
C
C
C
C
C
C
C
C
C
C
C
C
56
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
CD
Fig. S7. Effect of missing taxa on ancestral area reconstructions. (A) Phylogeny of tribes Isertieae and Cinchoneae, as estimated under the Bayesian analysis
(50% majority-rule consensus tree with all compatible groups added). J0–J9 indicate branches where missing taxa might be expected to attach, as suggested by
morphological evidence (e.g., morphology-based cladograms, accounts in taxonomic revisions); when no morphological evidence was available, the missing
taxon was attached to the most basal or most distal node of the genus. Node numbers as in Fig. S4. (B) Results from DIVA simulations after the addition of missing
taxa at each intersection node (J0–J9) outlined above. The distribution of the missing (hypothetical) taxon added is given after each intersection node, according
to the area codings in Fig. 1 (Inset) and based on the geographic bias estimated for each genus (Fig. S6). Ancestral area distributions differing from the original
reconstruction (no taxon addition) are marked in yellow.
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A)
Node 20
50
Node 31
Node 32
40
Node 33
Age (Ma)
Node 40
Node 42
30
Node 44
Node 45
Node 46
20
Node 52
Node 55
10
Node 56
All the above
combined
0
40
60
80
100
120
140
160
180
Percentage of taxa included
B)
Node
No.
20
31
32
33
40
42
44
45
46
52
55
56
Percentage of
taxa inc luded
Ages based on 1000 trees
(all ta xa incl uded)
30
40
60
80
100
Mean
46.5
39.0
20.1
9.25
24.3
15.8
14.3
13.5
6.45
3.63
12.7
10.9
48.9
44.1
22.6
10.7
26.7
17.8
15.7
14.7
6.95
4.09
13.8
11.6
49.0
40.2
22.7
12.6
25.3
16.7
14.8
14.1
6.92
4.30
13.1
11.1
50.3
41.9
24.3
12.8
22.3
17.9
15.8
14.9
7.16
4.59
13.9
11.4
51.3
44.5
25.1
13.7
28.0
18.6
16.8
15.7
9.08
4.94
13.8
10.9
51.3
45.2
25.0
13.1
28.6
19.0
17.2
16.2
9.00
5.19
14.4
11.8
95% Confidence Interval
Lower bound Upper bound
47.8
54.6
39.7
50.0
19.9
31.4
9.78
18.3
22.9
35.1
14.0
24.8
14.0
24.8
12.5
22.0
6.05
12.9
2.96
7.84
10.5
19.7
8.51
16.3
Fit of
trend l ine
R2
0.92
0.29
0.93
0.98
0.02
0.62
0.60
0.68
0.57
0.96
0.47
0.00
Fig. S8. Effect of missing taxa on divergence time estimations. (A) Ages of key nodes estimated from randomly reduced datasets (including 30%, 40%, 60%,
and 80% of the original number of species) and their expected development with the addition of missing taxa (projection curve calculated following a
second-degree logarithmic function). Node numbers as in Fig. S4. (B) Ground data for the diagram above, compared with age node statistics calculated from
the original dataset (all taxa included). Boxes marked in yellow fall outside the 95% confidence interval calculated from 1,000 Bayesian trees inferred from the
whole dataset. The last column shows the statistical fit of the trend line to the data points in the diagram.
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
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Table S1. Genera and number of species currently recognized in tribes Cinchoneae and Isertieae
Tribe
Cinchoneae
Isertieae
Genus
No. of species
Ciliosemina
Cinchona
Cinchonopsis
Joosia
Ladenbergia
Maguireocharis
Pimentelia
Remijia
Stilpnophyllum
Isertia
Kerianthera
2
23
1
11
34
1
1
41
4
14
1
A complete list of the species in each genus is given in Table S4.
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Table S2. Crown group ages of all nodes in the phylogeny
Consensus phylogram
Estimates based on 1000 Bayesian trees
95% credibility interval
Node
Age
Median Age
Lower
Upper
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
78.0
68.6
64.9
43.8
65.8
64.8
46.8
44.9
43.6
30.8
20.6
62.7
48.1
46.5
40.4
46.1
43.7
30.3
15.7
51.3
36.4
33.9
28.2
49.2
35.2
48.5
46.4
29.7
18.8
15.3
44.5
25.1
13.7
6.34
10.9
8.56
1.85
10.0
1.87
28.0
13.4
18.6
5.97
16.8
15.7
9.08
6.74
6.02
4.50
3.40
15.1
4.94
3.08
1.49
13.8
10.9
8.46
9.81
8.66
6.16
78.0
68.8
65.4
44.0
66.1
65.1
47.9
46.0
44.2
31.8
21.5
62.7
48.1
46.9
41.0
45.9
43.7
30.3
15.8
51.3
36.3
33.9
28.3
49.4
35.3
49.2
47.3
30.0
19.2
15.6
45.2
25.0
13.1
5.76
10.8
8.80
1.91
10.3
1.94
28.6
13.7
19.0
6.05
17.2
16.2
9.00
6.67
5.98
4.51
3.42
15.8
5.19
3.27
1.54
14.4
11.8
9.07
10.6
9.42
6.68
–
63.8
59.2
37.6
63.0
61.5
43.4
41.3
39.9
27.4
17.7
59.7
43.8
42.6
35.7
41.5
38.9
26.2
12.5
47.8
34.8
–
23.1
44.8
29.9
44.9
42.5
23.5
16.1
12.0
39.7
19.9
9.78
2.90
7.47
4.71
0.35
6.52
0.57
22.9
8.93
14.0
3.73
14.0
12.5
6.05
4.32
3.88
2.77
1.87
11.4
2.96
1.65
0.56
10.5
8.51
6.31
7.83
6.69
4.62
–
74.5
71.5
51.1
68.8
67.8
52.5
50.6
49.1
36.7
25.8
66.2
52.4
51.0
45.5
50.5
48.2
34.8
19.5
54.6
38.1
–
32.5
53.3
40.8
53.1
52.1
37.6
23.2
18.9
50.0
31.4
18.3
10.2
15.0
14.0
4.12
14.0
3.76
35.1
18.3
24.8
9.83
24.8
22.0
12.9
9.71
8.60
6.80
5.39
21.5
7.84
5.43
2.80
19.7
16.3
13.0
15.0
13.1
9.55
Statistics based on 2 calculations: Consensus phylogram refers to ages calculated from mean branch lengths of trees sampled by a Bayesian MCMC analysis
(n ⫽ 6,000); Estimates based on 1,000 Bayesian trees refer to median age estimates and 95% credibility intervals calculated by independently dating trees from
a random subsample of the Bayesian MCMC analysis (n ⫽ 1,000). Node numbers refer to Fig. S4.
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
17 of 20
Table S3. Sequences used in the phylogenetic and biogeographic analyses, together with source and GenBank accession numbers
Taxon
Outgroup
Gelsemium sempervirens (L.) J. St.-Hil.
Geniostoma rupestre J.R. Foster & G. Foster
Strychnos nux-vomica L.
Thevetia peruviana K. Schum.
Ingroup
Cephalanthus occidentalis L.
Chiococca alba (L.) Hitchc.
Cinchona calisaya Wedd.
C. macrocalyx Pav. ex DC.
C. mutisii Lamb.
C. officinalis L.
C. pitayensis Wedd.
C. pitayensis Wedd.
C. pubescens Vahl
C. pubescens Vahl
Cinchonopsis amazonica (Standl.) L. Andersson
Condaminea corymbosa (Ruiz & Pav.) DC.
Cosmibuena grandiflora (Ruiz & Pav.) Rusby
Cubanola domingensis (Britton) Aiello
Exostema lineatum (Vahl) Roem. & Schult.
Genipa americana L.
Guettarda speciosa L.
Hallea rubrostipulata (K. Schum.) J.-F. Leroy
Hamelia papillosa Urb.
Hymenodictyon floribundum Robinson
Isertia coccinea (Aubl.) J.F. Gmel.
I. laevis (Triana) Boom
I. haenkeana DC.
I. hypoleuca Benth.
I. parviflora Vahl
I. pittieri (Standl.) Standl.
I. rosea Spruce ex K. Schum.
I. spiciformis DC.
Joosia aequatoria Steyerm.
J. umbellifera H. Karst.
Kerianthera preclara Kirkbr.
Ladenbergia amazonensis Ducke
L. carua (Wedd.) Standl.
L. macrocarpa (Vahl) Klotzsch
L. oblongifolia (Mutis) L. Andersson
L. pavonii (Lamb.) Standl.
L. sp. (prob. nova)
Lasianthus batangensis K. Schum.
Luculia grandiflora Ghose
Mitriostigma axillare Hochst.
Mussaenda raiateensis J.W. Moore
Nauclea orientalis L.
Ophiorrhiza mungos L.
Paederia foetida L.
Pauridiantha sp.
Phyllis nobla L.
Posoqueria latifolia (Rudge) Roem. & Schult.
Psychotria kirkii Hiern
Remijia chelomaphylla G.A. Sullivan
R. macrocnemia (Mart.) Wedd.
R. pacimonica Standl.
R. pedunculata (H. Karst.) Flueck.
R. ulei K. Krause
Rovaeanthus suffrutescens (Brandeg.) Borhidi
Sabicea aspera Aubl.
Sipanea biflora (L.f.) Cham. & Schltdl.
Stilpnophyllum grandifolium L. Andersson
S. oellgaardii L. Andersson
Vangueria madagascariensis J.F. Gmel.
GenBank accession number
Origin
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
Colombia
Colombia
French Guiana
Colombia
Ecuador
French Guiana
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
GenBank
Voucher
ITS
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
Alzate et al. 203 (GB),
Andersson et al. 2173 (GB)
Andersson et al. 1969 (GB),
Andersson et al. 2099 (GB)
Hekker & Hekking 10.147 (GB)
Andersson et al. 1905 (GB
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
matK
—
—
—
—
Z70195
Z70194
Z70193
Z70188
—
—
AY538352
—
AY538353
AY538354
AY538355
—
AY538356
AY538377
AY538378
AY538379
AY538380
—
AY538381
AY538382
—
—
Z70197
AY538383
AY538384
AY538385
AY538386
AY538387
AY538388
AY538389
AY538390
AY538391
AY538392
AY538393
AY538394
—
—
—
—
—
—
AY538395
AY538396
AY538397
AY538398
AY538399
AY538400
AY538401
AY538402
AY538403
AY538404
Z701999
AY538405
AY538406
AY538407
AY538408
AY538409
AY538410
AY538411
AY538412
AY538413
AY538414
AY538415
AY538416
AY538417
AY538418
AY538419
AY538420
AY538421
AY538422
AY538423
AY538424
AY538357
—
—
—
—
—
—
—
—
—
AY538358
AY538359
DQ448607*
DQ448608*
DQ448609*
DQ448610*
DQ448611*
DQ448612*
AY538360
AY538361
AY538362
AY538363
AY538364
AY538365
AY538366
AY538367
AY538368
—
—
—
—
—
—
—
—
—
—
—
AY538369
AY538371
AY538372
AY538373
AY538374
—
—
—
AY538375
AY538376
—
rbcL
L14397
Z68828
L14410
X91773
X83629
L14394
AY538478
AY538479
—
AY538480
AY538481
—
—
X83630
AY538482
Y18713
AY538483
X83632
AY538484
Z68839
AY538485
AY538486
AY538487
AY538488
AY538489
AY538490
DQ448595*
DQ448596*
DQ448597*
DQ448598*
DQ448599*
DQ448600*
AY538491
AY538492
AY538493
AY538494
AY538495
AY538496
AY538497
Z68801
AY538498
AY538499
X83648
X83650
AY538500
AY538501
X83656
AF332373
AY538502
Z68814
Z68850
X83663
AY538503
AY538504
AY538505
AY538506
AY538507
X83665
AY538508
AY538509
AY538510
AY538511
X83670
rps16
trnL–F
AF004092
—
AF004094
—
AF159696
—
AF102484
—
AF004033
AF004034
AF242927
AF538425
AY538426
AY538427
—
AF242928
—
AF004035
AY538428
AF004039
AF242929
AF004044
AF242944
AF200997
AF246924
AF538429
AF004053
AF004058
AY538430
AY538431
DQ448601*
DQ448602*
DQ448603*
DQ448604*
DQ448605*
DQ448606*
AY538432
AY538433
AF242970
AY538434
AY538435
AF242971
AY538436
AY538437
AY538438
AY538439
AF242974
AF201006
AF242983
AY538440
AF004064
AF004065
AF004068
AF003613
AF242998
AF410728
AY538441
AY538442
AY538443
AY538444
AY538445
AF243003
AF004079
AF004085
AY538446
AF243026
AF243033
AF152692
AF102400
AY538447
AY538448
AY538449
AY538450
—
AF152684
AY538451
—
AY538452
AF102406
AF152686
AF152701
AF152698
AF201045
AF152725
AY538453
AF102439
AY538454
AY538455
AY538456
—
—
—
—
—
—
AY538457
AY538458
AY538459
AY538460
AY538461
AF152683
AF538462
AY538463
AY538464
AY538465
AF102453
AF201054
AY538466
AY538467
AF152610
AF152619
AF102467
AY538468
AF152680
AY538469
AY538470
AY538471
AY538472
AY538473
AY538474
AF152738
AY538475
AF152675
AY538476
AY538477
AF152654
Data on origin and voucher are given only for sequences added in this study (indicated by an asterisk).
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
18 of 20
Table S4. Checklist of species and genera in the sister tribes Cinchoneae and Isertieae, their distribution, and number of collections
verified in recent taxonomic revisions (see SI Text for references)
Species count in tribes
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
Species count in genera
1
2
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
1
1
2
3
4
5
6
7
8
9
10
11
12
13
14
1
2
3
4
5
6
7
8
9
10
11
1
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
Genus
Ciliosemina
Ciliosemina
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchona
Cinchonopsis
Isertia
Isertia
Isertia
Isertia
Isertia
Isertia
Isertia
Isertia
Isertia
Isertia
Isertia
Isertia
Isertia
Isertia
Joosia
Joosia
Joosia
Joosia
Joosia
Joosia
Joosia
Joosia
Joosia
Joosia
Joosia
Kerianthera
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Species
pedunculata
purdieana
antioquiae
asperifolia
barbacoensis
calisaya
capuli
fruticosa
glandulifera
hirsuta
krauseana
lancifolia
lucumifolia
macrocalyx
micrantha
mutisii
nitida
officinalis
parabolica
pitayensis
pubescens
pyrifolia
rugosa
scrobiculata
villosa
amazonica
coccinea
haenkeana
hypoleuca
krausei
laevis
longifolia
parviflora
pittieri
reticulata
rosea
scorpioides
spiciformis
verrucosa
wilhelminensis
aequatoria
dichotoma
dielsiana
longisepala
macrocalyx
multiflora
obtusa
oligantha
pulcherrima
standleyana
umbellifera
preclara
acutifolia
amazonensis
brenesii
bullata
buntingii
carua
chapadensis
cujabensis
discolor
dwyeri
epiphytica
ferruginea
graciliflora
heterophylla
hexandra
klugii
lambertiana
laurifolia
Distribution
No. of collections
CF
F
C
D
CE
D
C
C
D
D
D
C
C
CD
CD
C
D
C
CD
C
ACD
D
C
C
C
F
F
ABCEF
ACFG
D
ACDE
F
FG
EC
D
F
A
FG
F
G
C
D
C
C
C
D
C
C
CD
C
ACDF
F
D
DFG
A
D
C
D
H
H
D
A
C
D
DFH
AC
H
D
FG
A
>6
2
⬎6
2
⬎6
>6
⬎6
2
5 or 6
4
5 or 6
⬎6
⬎6
>6
⬎6
>6
5
>6
6
>6
>6
3
⬎6
1 (type only)
2
>6
>6
>6
>6
1 (type only)
>6
⬎6
>6
>6
5
>6
5
>6
5
3
2
⬎6
⬎6
2
1 (type only)
1 (type only)
1 (type only)
1 (type only)
⬎6
3
>6
4
4
>6
⬎6
2
3
>6
1 (type only)
⬎6
6
⬎6
1 (type only)
1 (type only)
⬎6
⬎6
⬎6
1 (type only)
⬎6
3
19 of 20
Species count in tribes
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
Species count in genera
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
1
1
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
1
2
3
4
Genus
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Ladenbergia
Maguireocharis
Pimentelia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Remijia
Stilpnophyllum
Stilpnophyllum
Stilpnophyllum
Stilpnophyllum
Species
lehmanniana
macrocarpa
magdalenae
moritziana
muzonensis
nubigena
oblongifolia
obovata
paraensis
pauciflora
pavonii
pittieri
riveroana
rubiginosa
stenocarpa
undata
neblinae
glomerata
amazonica
amphithrix
aracamuniensis
argentea
asperula
berryi
bracteata
chelomaphylla
delascioi
densiflora
duckei
ferruginea
firmula
globosa
glomerata
hispida
involucrata
leiocalyx
longifolia
macrocnemia
macrophylla
maguirei
marahuacensis
megistocaula
morilloi
pacimonica
paniculata
peruviana
physophora
pilosinervula
prismatostylis
reducta
roraimae
sessilis
sipapoensis
steyermarkii
tenuiflora
trianae
ulei
vaupesiana
wurdackii
grandifolium
lineatum
oellgaardii
revolutum
Distribution
No. of collections
C
C
C
C
CEF
A
CDEFG
C
F
C
CE
C
C
CE
C
C
G
CD
F
F
G
FG
F
F
F
CD
F or G
G
F
H
FG
G
F
F
F
E
F
F
C
G
F
F
F
F
H
F
F
G
C
F
FG
F
G
G
F
F
F
F
F
C
D
C
C
3
>6
⬎6
⬎6
⬎6
5
>6
3
1 (type only)
1 (type only)
>6
⬎6
⬎6
2
⬎6
⬎6
2
⬎6
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
not revised
1 (type only)
1 (type only)
2
1 (type only)
Species in bold type were included in the molecular phylogeny. Distribution codings are according to Fig. 1 Inset.
Antonelli et al. www.pnas.org/cgi/content/short/0811421106
20 of 20
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