Flora 281 (2021) 151870

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Flora
journal homepage: www.elsevier.com/locate/flora

Floral synorganization in acmantheroid clade suggests hypotheses to

explain elaiophore suppression in Malpighiaceae
Stéphani Karoline Vasconcelos Bonifácio a, *, Rafael Felipe de Almeida b,
André Márcio Araújo Amorim c, Denise Maria Trombert Oliveira a
a
PlantSeR Lab, Departamento de Botânica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil
b
Independent researcher, Brazil
c
Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz, Ilhéus, Bahia, Brazil

A R T I C L E I N F O A B S T R A C T

Editor: Louis Philippe Ronse De Craene Elaiophores in angiosperms first appeared in Malpighiaceae and are fundamental for this family’s sexual
reproduction. However, they are absent in some lineages. The complete loss of calyx oil glands is an important
Keywords: deviation from the floral conservatism of the Malpighiaceae. Although the literature points to ontogenetic
Acmanthereae processes involved in the loss of anterior sepal glands in some lineages, the eglandular calyx remains unex­
Calyx glands
plained. Thus, understanding how flowers are losing the main structure responsible for conserving their floral
Floral vasculature
architecture is important to comprehend the processes that drive floral evolution in Malpighiaceae. We selected
Hypanthium
Malpighiales one species from each genus of the acmantheroid clade, which is the unique lineage with all the main character
states related to the occurrence of elaiophores. Floral buds near anthesis were processed using usual techniques
for light microscopy. The calyx vasculature is variable and forms vascular complexes shared between sepals,
petals, and, in some species, stamens. For the first time, we describe the occurrence of a hypanthium in Mal­
pighiaceae, indicating different levels of floral synorganization. Floral synorganization appears to be the key to
understand how specific neotropical lineages broke the floral conservatism in Malpighiaceae with eglandular
calyces. The presence of different vascular complexes between perianth parts allows us to infer the group’s floral
evolution. Thus, we propose two hypotheses that explain the absence of the elaiophores and indicate the ten-
elaiophore condition as plesiomorphic for Malpighiaceae.

1. Introduction
Oil as a floral reward did not exist in angiosperms until the appear­
ance of Malpighiaceae around 80 Ma. (Renner and Schaefer, 2010). In
the family, the main elaiophores (oil secretory structures) are inserted
on the calyx and considered a morphological synapomorphy (Vogel,
1974; Davis and Anderson, 2010). These glands play an important role
in the sexual reproduction of neotropical species since some of these taxa
depend on pollinators due to herkogamy, protogyny, and the presence of
a cuticle on the stigma (Sigrist and Sazima, 2004). The role of calyx
glands (which are mentioned in this text simply as elaiophores) is so
vital in the attraction of oil-collecting bees that the floral morphology
among neotropical species hardly varies, and its conservatism is attrib­
uted to the mutualistic relationship with these insects (Vogel, 1974,
1990; Anderson, 1979; Simpson and Neff, 1981; Davis et al., 2014).
The presence of elaiophores is a synapomorphy in Malpighiaceae
(Anderson, 1990a). However, the number of elaiophores per flower is
variable, and their total absence can be noticed at every taxonomic level
(Anderson, 1990a): at population, genus (Anderson, 1979; 1990a;
Cappellari et al., 2011) and individual (Sazima and Sazima, 1989; Car­
valho et al., 2005) levels. The presence of an eglandular calyx is
considered homoplastic and appeared at least 14 times in the family,
including seven times only in the Neotropics (i.e., in Coleostachys A.Juss,
Galphimia Cav., Echinopterys A.Juss., Ectopopterys W.R.Anderson, Lasio­
carpus Liebm. + Ptilochaeta Turcz clade, Psychopterys W.R.Anderson & S.
Corso and Thryallis Mart.) (Anderson et al., 2006; Renner and Schaefer,
2010; Davis et al., 2014). Phylogenetic relationships can be seen in
Fig. 1. The process appears to be ongoing in the family since the total
number of elaiophores can vary from zero to ten among the genera
(Anderson et al., 2006; Guesdon et al., 2019), as well as at species and
specimen level (Vogel, 1990).
Additionally, the presence of eight elaiophores is the most recurrent
in the Neotropics, and, in this case, the flower during anthesis has an
eglandular anterior sepal (Anderson, 1990a; Anderson et al., 2006).

* Corresponding author.

https://doi.org/10.1016/j.flora.2021.151870
Received 18 November 2020; Received in revised form 15 May 2021; Accepted 1 June 2021
Available online 7 June 2021
0367-2530/© 2021 Elsevier GmbH. All rights reserved.
S.K.V. Bonifácio et al.
(caption on next column)
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Flora 281 (2021) 151870
Fig. 1. Summary of major clades of Malpighiaceae and its sister-group Elati­
naceae (dashed line and bold type) showing acmantheroid clade relationship
with other clades (adapted from Davis et al., 2001; Davis and Anderson, 2010,
including the new genus Andersoniodoxa from Davis et al., 2020a modified by
Davis et al., 2020b and the reference to Elatinaceae from Davis and Chase,
2004). The numbers in front of the clades represent species with described
floral vasculature, studied by Souto and Oliveira (2013)1 and A.C.M.P. Mello, R.
F. Almeida, A.M.A. Amorim, and D.M.T. Oliveira (personal communication)2.
asterisks indicate the occurrence of eglandular species (Anderson et al., 2006).
From an ecological perspective, the suppression of the elaiophores on
the anterior sepal occurred because oil-collecting bees could not access
them and, thus, these glands were lost over time (Sazima and Sazima,
1989; Cappellari et al., 2011). From an anatomic point of view, Souto
and Oliveira (2013) describe the floral vasculature of some Malpighia­
ceae, and their observations indicate two distinct processes that resulted
in the eglandular sepal: in Mascagnia cordifolia (A.Juss.) Griseb. the
vasculature indicates connation between elaiophores of adjacent sepals;
in Tetrapterys chamaecerasifolia A.Juss. and Janusia mediterranea (Vell.)
W.R.Anderson, there is a numerical reduction since there are no signs of
the vasculature of the suppressed elaiophores.
In contrast, the only ecological explanation for the complete sup­
pression of elaiophores is deceit pollination, where eglandular morphs
attract pollinators by mimicking glandular flowers (Sazima and Sazima,
1989; Sigrist and Sazima, 2004; Carvalho et al., 2005; Cappellari et al.,
2011). However, no ontogenetic investigations point out the processes
responsible for the complete loss of elaiophores. Besides, the clades that
have information available about floral vasculature are derived in the
Malpighiaceae, so it is not possible to infer if connation or reduction is
related to the suppression in all eglandular neotropical genera.
In this scenario, the acmantheroid clade sensu Davis and Anderson
2010 (= tribe Acmanthereae W.R.Anderson of Anderson, 1977) is an
excellent model to study the processes related to the loss of calyx glands
in Malpighiaceae. The clade comprises three genera (Davis and Ander­
son, 2010) with 0–10-glandular representatives (see a summary of major
clades in Fig. 1): Coleostachys is monospecific, eglandular (Almeida and
Hall, 2016), and the sister to the other genera; Acmanthera Griseb.
comprises seven species always has ten elaiophores (Anderson, 1975,
1980a, 1990b; Farroñay et al., 2019); and Pterandra A.Juss., with 15
species, has in the same species different morphs with ten glands, glands
irregular in number and format, or no glands (Anderson, 1997, 2005a).
Further, the acmantheroid clade is one of the first lineages that diverged
in Malpighiaceae (Davis and Anderson, 2010; Davis et al., 2020), and
investigating the floral vasculature of its representatives may clarify the
processes that culminated in the absence of the glands. These combined
characteristics make the acmantheroid clade unique within the family.
Revealing the processes involved in the loss of calyx glands is a
significant step towards understanding the floral evolution of Malpigh­
iaceae, especially the mutualism between the family and oil-collecting
bees, which can be lost with the suppression of the glands. Informa­
tion about the suppression of glands can also help interpret relationships
between subfamilies and tribes, which are polyphyletic according to the
most recent phylogeny (Davis and Anderson, 2010). Considering that
our subject is the suppression of the elaiophores, this work aims to
characterize the calyx vasculature in the acmantheroid clade and pro­
pose hypotheses that can explain the complete suppression of the
elaiophores in some Malpighiaceae.
2. Material and methods
We selected one species per genus (or the only species, i.e., Coleos­
tachys genipifolia A.Juss.) in the acmantheroid clade, and three in­
dividuals were sampled per species. For Pterandra pyroidea A.Juss., we
sampled individuals with no elaiophores (eglandular morph) and with
elaiophores (glandular morph). The material consisted of floral buds
near anthesis taken from herbarium specimens (Table 1). We submitted
S.K.V. Bonifácio et al. Flora 281 (2021) 151870

Table 1 comprising traces that originate basipetally and irrigate the distal por­
Species and specimens of the genera of the acmantheroid clade sampled in this tions of the receptacle and elaiophores (Fig. 2E-F). Since there are
work. basipetal, branched traces, the xylem and phloem are inverted, and the
Species Glandular Voucher Collection locality in internal xylem in acropetal branches becomes external in basipetal
condition of the states of Brazil branches (Fig. 2G).
calyx The sepal-petal-antipetalous stamen complexes are emitted in the
Coleostachys Eglandular Silva Jr 59 Parauapebas, Pará regions between the sepals (Fig. 2D). These vascular complexes are so
genipifolia A.Juss. (MG) named because they ramify and emit the lateral sepal, petal, and anti­
Vidal 712 Parauapebas, Pará
petalous stamen traces. The antisepalous stamen traces (et; yellow in the
(CEPEC)
Sales 02 Parauapebas, Pará figures) are emitted from the vascular cylinder (Fig. 2E), while each
(INPA) sepal-petal-antipetalous stamen complex irrigates a sepal twice (Fig. 2B,
Acmanthera latifolia Glandular Martinelli Bonfim, Roraima E), one petal and one antipetalous stamen. First, the complex migrates to
(A.Juss.) Griseb. 17688 the receptacle periphery, where it branches, and its ends irrigate the
(CEPEC)
Amorim 8575 Barcelos, Amazonas
elaiophores (Fig. 2H). Above the central portion of the complex, the
(CEPEC) sepal trace is externally differentiated and irrigates the glands again,
Anderson Barcelos, Amazonas followed by the petal trace (pt; green in the figures) and, internally, the
13771 antipetalous stamen trace (tt; yellow in the figures) (Fig. 2I). Even after
(CEPEC)
the differentiation of the vascular traces at the height after this region,
Pterandra pyroidea Glandular Silva 73 Brasília, Distrito
A. Juss. (CEPEC) Federal the calyx and corolla remain adnate until the anthers’ height. However,
Romero 4442 São Roque de Minas, antipetalous stamens are separate (Fig. 2J).
(CEPEC) Minas Gerais Thus, each sepal receives three vascular traces: a middle trace,
Fonseca 3481 Mimoso de Goiás, coming directly from the stele, and two lateral traces, from the sepal-
(CEPEC) Goiás
petal-antipetalous stamen complexes adjacent to each sepal. The
Pterandra pyroidea Eglandular Duarte 2807 Serra do Salitre,
A. Juss. (CEPEC) Minas Gerais petals and stamens, in turn, receive one trace each.
Francener Pirenópolis, Goiás
1139 (CEPEC) 3.2. Pterandra pyroidea
Pereira 709 Brasília, Distrito
(CEPEC) Federal
The elaiophores in Pterandra pyroidea A.Juss., when present (the
species has glandular and eglandular morphs), are in the lower part of
the samples to an herborization reversal procedure (Smith and Smith, the sepals, fused to the receptacle, and are vascularized only by vascular
1942, modified by Mello et al., 2019), dehydrated them in an increasing complexes that also irrigate the calyx (Fig. 3A–B).
ethanol series, and stored them in 70% ethanol. In the glandular morph, the number of elaiophores varied from six to
The samples were then embedded in Leica (2-hydroxyethyl)-meth­ nine, irregularly distributed among the samples. Before the emission of
acrylate (Paiva et al., 2011) and sectioned (10 µm thick) with a Zeiss the traces, the elaiophores are already vascularized, as well as the small
Hyrax M40 rotary microtome to obtain transversal series. Subsequently, extension of the receptacle fused to them (Fig. 3C). The calyx is vascu­
the sections were stained with 0.05% toluidine blue in a pH 4.7 acetate larized only by traces from vascular complexes: in the portions corre­
buffer (O’Brien et al., 1964, modified), counterstained with ruthenium sponding to the middle region of the sepals, five median sepal traces
red (Gonçalves-Souza et al., 2018), and mounted in Entellan. modified into sepal-petal complexes (sp; pink in the figures), and be­
The images were obtained using an LC20 camera coupled to an tween the sepals, five sepal-petal-antipetalous stamen complexes are
Olympus CX41microscope, joined and aligned with the software Image emitted (ss; light blue in the figures) (Fig. 3D). The ends of both com­
Composite Editor 2015, and edited with Corel PHOTO-PAINTTM 2020 plexes unite and migrate to the elaiophores basipetally, while anti­
software. The figures and illustrations of the vasculature of the calyx and sepalous stamen traces (et; yellow in the figures) are emitted from the
other directly associated floral whorls were produced from photomi­ vascular cylinder (Fig. 3E). These vascular traces irrigate the elaiophores
crographs with the vector program CorelDRAWTM Graphics Suite 2020. only once. At the ovary base, the lateral sepal, median petal, and anti­
The terminology adopted to describe the floral vasculature follows petalous stamen traces separate from the complexes (Fig. 3E).
Puri (1951), Souto and Oliveira (2013), and A.C.M.P. Mello, R.F. In the eglandular morph, the vasculature pattern is similar to that
Almeida, A.M.A. Amorim, and D.M.T. Oliveira (personal communica­ described above. The distal receptacular region is vascularized in the
tion). In this sense, we have adopted the term vascular complexes to position corresponding to the region of the elaiophores (Fig. 3F–H). Like
designate those vascular traces that, after branching, supply more than the glandular morph, five median sepal traces modified into sepal-petal
one floral whorl simultaneously, e.g., calyx and corolla. complexes and five sepal-petal-antipetalous stamen complexes are
emitted (Fig. 3I). All the vascular complexes fuse at the level of the base
3. Results of the ovary, where the traces start to separate from the complexes and
antisepalous stamen traces are emitted from the vascular cylinder
3.1. Acmanthera latifolia (Fig. 3J).
In both morphs, each petal is vascularized from three vascular traces
In Acmanthera latifolia (A.Juss.) Griseb., the ten elaiophores are fused (Fig. 4A, E, I): the median trace petal, from the sepal-petal-antipetalous
to the receptacle and vascularized by median sepal traces and vascular stamen complexes, and two lateral petal traces. These lateral traces
complexes emitted from the vascular cylinder (Fig. 2A–B). originate from the median sepal traces modified into sepal-petal com­
Before the emission of a trace, the peripheral receptacular region, plexes and have variable patterns between the petals of the same flower,
and more distant portions of the elaiophores are vascularized (Fig. 2C). independently of the occurrence of elaiophores.
In turn, the calyx is vascularized from five median sepal traces (st; red in In total, we observed three different patterns of emission of vascular
the figures) and five sepal-petal-antipetalous stamen complexes (ss; light traces towards petals in P. pyroidea. In the first, the end of the adjacent
blue in the figures) (Fig. 2D). sepal-petal complex bifurcates, and the branches migrate towards the
The median sepal traces are in the middle of the sepals (Fig. 2D–E). side of the same petal (Fig. 4A–D). In the second case, part of the end of
When they migrate to the receptacular periphery, they branch and end the sepal-petal complex fuses to the sepal-petal-antipetalous stamen
complex, and then the three petal traces are emitted (Fig. 4E–H). In the

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S.K.V. Bonifácio et al. Flora 281 (2021) 151870

Fig. 2. Calyx and associated whorls vasculature pattern in floral buds near anthesis of Acmanthera latifolia. Diagrams (A-B, E) and photomicrographs of transversal
sections (C–D, F–J). The colors represent sepal traces (red), petal traces (green), sepal-petal-antipetalous stamen complexes (light blue), stamen traces (yellow) and
the vascular cylinder and its remnants for gynoecium (black). A. Diagram of median longitudinal reconstruction of the floral bud showing the vascular pattern; the
letters following the dashed lines indicate the approximate position of the corresponding images. B. Representation of a floral bud highlighting the origin of the
vasculature of a calyx gland; the dashed lines indicate basipetal traces. C–E. General views of the receptacle showing the emission of the vascular traces and
complexes, highlighted by the areas with the colors indicated above; the ellipses in C delimit the regions corresponding to each sepal; in E and F, the dashed circles
highlight the basipetal traces. F. Detail of the distribution of the basipetal traces; near the center of the image, note that median sepal trace (st) is oriented acropetally,
so xylem is inner to phloem. G. Detail of a basipetal trace; note the position of the xylem vessel elements external to the phloem. H–I. Sequence of the branching
pattern of a sepal-petal-antipetalous stamen complex. J. Region above H–I sequence, showing the petal trace originating from the sepal-petal-antipetalous stamen
complex. arrow, branched traces from basipetal traces; asterisk, calyx gland; as, anterior sepal; co, corolla; et, antisepalous stamen traces; gl, calyx gland; ph, phloem;
pt, petal trace; sa, stamen; se, sepal; ss, sepal-petal-antipetalous stamen complex; st, median sepal trace; tt, antipetalous stamen trace; xy, xylem.

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S.K.V. Bonifácio et al. Flora 281 (2021) 151870

Fig. 3. Calyx vasculature and associated whorls of floral buds near anthesis of Pterandra pyroidea. Glandular (A–E) and eglandular (F–J) morphs. Diagrams (A-B, D, F-
G, I) and photomicrographs of transversal sections (C, E, H, J). The colors represent sepal traces (red), sepal-petal complexes (pink), petal traces (green), sepal-petal-
antipetalous stamen complexes (light blue), stamen traces (yellow), carpel traces (dark blue) and the vascular cylinder and its remnants for gynoecium (black). A, F.
Diagrams of median longitudinal reconstruction of floral buds; the letters following the dashed lines indicate the corresponding following images. B, G. Repre­
sentation of floral buds; note the vasculature in the marginal region of two adjacent sepals; the dashed lines indicate basipetal traces. C–E, H–J. General views of the
receptacle showing the emission of vascular traces and complexes highlighted by the areas shaded with the colors indicated above; the ellipses in E and J indicate the
basipetal traces. arrow, branched traces from basipetal traces; asterisk, calyx glands; co, corolla; et, antisepalous stamen traces; gl, calyx glands; pt, petal trace; se,
sepal; sp, median sepal trace modified into sepal-petal complex; st, sepal trace; ss, sepal-petal-antipetalous stamen complex; tt, antipetalous stamen trace.

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S.K.V. Bonifácio et al. Flora 281 (2021) 151870

Fig. 4. Origin of the lateral petal traces of Pterandra pyroidea. The diagrams to the left summarize the patterns corresponding to the sequences to the right; the dashed
lines represent a fusion between the complexes. The colors represent sepal-petal complexes (pink), petal traces (green), sepal-petal-antipetalous stamen complexes
(light blue) and the vascular cylinder (black). The photomicrographs were taken from the region below the beginning of the hypanthium. A–D. Pattern involving ends
of the sepal-petal complex. E–H. Pattern involving a sepal-petal complex and a sepal-petal-antipetalous stamen complex. I–L. Pattern involving the two sepal-petal
complexes adjacent to the sepal-petal-antipetalous stamen complex. M–O. Sequence of petals receiving the three traces until they separate. arrow, indicates the origin
of the lateral petal traces; fi, filament; hy, hypanthium; pe, petal; pt, petal trace; se, sepal; sp, median sepal trace modified into sepal-petal complex; ss, sepal-petal-
antipetalous stamen complex; tt, stamen trace.

third pattern, the lateral ends come from only one sepal-petal complex;
however, there is a fusion between branches of the two adjacent sepal-
petal complexes to the sepal-petal-antipetalous complex (Fig. 4I–L).
Acropetally, the petals and filaments separate, and each petal maintains
the three vascular bundles (Fig. 4M–O).
In summary, each sepal and petal receive three vascular traces, and
each stamen receives only one trace.
3.3. Coleostachys genipifolia
The eglandular calyx is vascularized by median sepal traces and
traces emitted by the sepal-petal complexes (Fig. 5A–B). The basipetal
traces are present and originated exclusively from the sepal-petal com­
plexes (sp; pink in the figures) (Fig. 5B) that peripherally irrigate the
calyx.

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S.K.V. Bonifácio et al. Flora 281 (2021) 151870

Fig. 5. Calyx vasculature and associated whorls of floral buds near anthesis of Coleostachys genipifolia. Diagrams (A-C, F) and photomicrographs of transversal
sections (D–E, G–J). The colors represent sepal traces (red), sepal-petal complexes (pink), petal traces (green), stamen traces (yellow), carpel traces (dark blue) and
the vascular cylinder and its remnants for gynoecium (black). A. Diagram of median longitudinal reconstruction of the floral bud showing the distribution pattern of
the vascular traces; the letters following the dashed lines indicate the approximate position of the corresponding images. B. Representation of a floral bud; note the
vasculature in the marginal region of two adjacent sepals; the dashed lines indicate basipetal traces. C–E. General views of the receptacle showing the emission of the
vascular traces and complexes. F. General view of hypanthium at the height of the already individualized ovary. G–J. Sequence of emission and branching of a sepal-
petal complex. arrow, branches from sepal-petal complex; as, anterior sepal; co, corolla; et, antisepalous stamen traces; gy, gynoecium; se, sepal; sp, sepal-petal
complex; st, median sepal trace; tt, antipetalous stamen trace.

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S.K.V. Bonifácio et al. Flora 281 (2021) 151870

In the periphery of the receptacle, there are vascular traces before the
branching of the five median sepal traces (st; red in the figures) and five
sepal-petal complexes (Fig. 5C–D). The emission of these five traces and
five complexes is simultaneous and immediately followed by the emis­
sion of all stamen traces (Fig. 5E). Each median sepal trace branches
before irrigating the calyx, so the middle sepal trace is indistinct
(Fig. 5F).
One sepal-petal complex migrates to one of the adjacent sepals and
then bifurcates (Fig. 5G); at this point, although not emitted yet, the
traces originating from the other side of the bifurcation are already
visible. Above this, the branch to the other sepal is emitted (Fig. 5H),
and, in this plane, the external traces fuse. Then, the newly emitted trace
migrates in the direction of the corresponding external trace (Fig. 5I). In
the end, four traces originate from one sepal-petal complex (Fig. 5J),
which acropetally and basipetally bifurcate.

3.4. Floral synorganization in the acmantheroid clade

All the species analyzed form a hypanthium (sensu Gifford and Fos­
ter, 1989) that involves adnation between the perianth parts and the
androecium. The separation between the floral whorls occurs at heights
different from than the emission of the vascular traces. In A. latifolia
(Fig. 6A-B) and P. pyroidea (Fig. 6C-D), the fusion extends from the level
where the gynoecium is visible, when vascular complexes are already
differentiated, to the level of petals and stamens. In C. genipifolia, the
hypanthium extends from gynoecium is visible, through vascular com­
plexes emission (Fig. 6E), after stamen and then petals separation
(Fig. 6F).

4. Discussion

Calyx vasculature in Malpighiaceae is complex and variable between

genera, species, and floral morphotypes, including processes of fusion
that involve the formation of vascular complexes. Unlike what has been
reported for species of more recent lineages of Malpighiaceae – tetrap­
teroids, stigmaphylloids (Souto and Oliveira, 2013) and malpighioids
(Souto and Oliveira, 2013; A.C.M.P. Mello, R.F. Almeida, A.M.A.
Amorim and D.M.T. Oliveira, personal communication) –, the calyx
vasculature in the acmantheroid clade is not uniform. In this sense, the
tribes Acmanthereae, Byrsonimeae W.R.Anderson, and Galphimieae
Nied. might constitute another evolutionary line since they represent the
first large lineage of Malpighiaceae and is the sister group to the rest of
the family (Anderson et al., 2006; Davis and Anderson, 2010).

4.1. Floral synorganization in Malpighiaceae

Our data indicate that the floral synorganization is constant in the

acmantheroid clade and possibly a tendency in Malpighiaceae, a family
in which several fusion processes occur and can result in the numerical
reduction of floral parts, which is recurrent in several genera (Anderson
et al., 2006). Previous works describe sepal-petal complexes in the
family (Souto and Oliveira, 2013; A.C.M.P. Mello, R.F. Almeida, A.M.A.
Amorim, and D.M.T. Oliveira, personal communication), but the present
is the first work to report sepal-petal-antipetalous stamen complexes.
The vascular fusion represents the last step in the fusion process, which
starts from the outside to the inside of each organ (Eames and MacDa­
niels, 1947); anatomically, the process starts from epidermal, then
ground tissue, and lastly at the vascular level. Thus, the origin of the
Fig. 6. General view of the hypanthium of floral buds near anthesis of
vascular traces shared between the calyx and corolla, and among the
Acmanthera latifolia (A-B), Pterandra pyroidea (C-D), Coleostachys genipifolia (E-
calyx, corolla, and androecium, in a basal clade of Malpighiaceae in­ F); note the separation of the gynoecium and filaments in A–F. as, anterior
dicates that the adnation between these floral whorls is phylogenetically sepal; et, antisepalous stamen traces; gy, gynoecium; fi, filament; hy, hypan­
old. thium; pp, posterior petal; pt, petal trace (green ellipse); sp, sepal-petal complex
The sepal-petal complex observed in both the morphs of P. pyroidea (pink ellipse); ss, sepal-petal-antipetalous stamen complex (blue ellipse); tt,
differs from those described for C. genipifolia and descriptions of other antipetalous stamen trace (yellow ellipse).
groups in Malpighiaceae (Souto and Oliveira, 2013; A.C.M.P. Mello, R.F.
Almeida, A.M.A. Amorim, and D.M.T. Oliveira, personal

8
S.K.V. Bonifácio et al.
communication). Although the complexes irrigate the calyx and corolla,
the branching pattern, position, and bundles from which they originate
are different. The petals of P. pyroidea receive three traces, of which the
median comes from sepal-petal-antipetalous stamen complexes, and the
lateral traces originate from sepal-petal complexes. Thus, the entire
perianth is vascularized from vascular complexes in this species, which,
as far as we know, has not been previously reported for the
Malpighiaceae.
The data here confirm how the development of calyx and corolla are
correlated in the flowers of Malpighiaceae. This relation is evident when
considering the floral symmetry, where the bilateral corolla in the
family is associated with the presence of elaiophores (Davis and
Anderson, 2010; Zhang et al., 2010). In the species analyzed here, in
which the glandular or eglandular state is fixed, there is no variation in
the corolla symmetry plane. Acmanthera (glandular) and Coleostachys
(eglandular) have a bilateral corolla, while Pterandra (genera where
both conditions occur) can be nearly radial (Anderson et al., 2006).
Thus, parallel to the process of establishing the eglandular condition, we
have to consider the change in the plane of the corolla symmetry, since
the vasculature highlights the close relationship between these whorls.
Further, the symmetry plane change could be related to a generalized
pollination system, considering that specialist pollinators tend to be
attracted to zygomorphic flowers, unlike generalist pollinators (Gong
and Huang, 2009).
4.2. Origin of the vasculature of the elaiophores in acmantheroid clade
The presence of similar basipetal traces that irrigate elaiophores even
in eglandular flowers, described in this study, and by Souto and Oliveira
(2013) and A.C.M.P. Mello, R.F. Almeida, A.M.A. Amorim, and D.M.T.
Oliveira (personal communication), reinforces Anderson’s hypothesis
(1979) that the glandular condition is a precursor of the eglandular
condition. The presence of the 10-glandular calyx seems to be the ple­
siomorphic condition since the basipetal traces are associated with all
five sepals. Therefore, the presence of basipetal traces in eglandular
flowers can be understood as vestigial from suppressed glands. Besides,
genera with 10-glandular calyx occupy the first branches of Malpigh­
iaceae (Davis and Anderson, 2010). Thus, the basipetal traces can be
recognized as a phylogenetic signal for the presence of ancestral
elaiophores.
Comparing the acmantheroid genera to each other, and considering
the 10-glandular condition as plesiomorphic, Acmanthera latifolia retain
character states from the ancestral condition: basipetal traces irrigate
elaiophores twice, and there is no additional vascular fusion between
adjacent sepals. In addition, the shared origin of the lateral sepal traces
between adjacent sepals, indicates a primary connation between sepals
in the ancestor of the clade. Pterandra pyroidea, on the other hand, does
not present an additional fusion between adjacent sepals, but its elaio­
phores are irrigated only once by the basipetal traces. Coleostachys
genipifolia, even though eglandular, presents the basipetal traces irri­
gating the region below the sepals twice (in the position corresponding
to the elaiophores in glandular species), but, differently from the other
genera, shows an additional (secondary) connation between adjacent
sepals. Thus, the vascular fusion between adjacent sepals and a reduc­
tion in the number of basipetal traces would be derived conditions,
resulting in floral morphology changes.
The number of traces that irrigates the elaiophores and their origin
also vary between the glandular species analyzed here. In A. latifolia, the
elaiophores are irrigated by two types of traces: those from median sepal
traces and those from sepal-petal-antipetalous stamen complexes. On
the other hand, the elaiophores of P. pyroidea are vascularized only by
traces originating from the fusion between sepal-petal-antipetalous
stamen complexes and ends of lateral sepal traces from median sepal
traces modified into sepal-petal complexes. Thus, in this species, the
basipetal traces are emitted only in the region between sepals, irrigating
them once and then branching higher up.
9
Flora 281 (2021) 151870
This reduction in the number of traces emitted to the elaiophores
indicates the vascular adaptation to reducing the gland tissue. The
presence of these traces even on the eglandular morph of P. pyroidea
indicates that this reduction is ongoing in the species. Thus, the nu­
merical reduction of traces allows the numerical suppression of elaio­
phores, which seems to be more parsimonious (involving fewer
evolutionary steps). As an organ decreases in size, less vasculature it
should receive.
4.3. Emission of three traces per petal in Pterandra pyroidea
Although uncommon, the emission of three traces per petal, first
reported here for Malpighiaceae, is also mentioned for other families of
Malpighiales: Irvingiaceae and Caryocaraceae (Matthews and Endress,
2011; Endress et al., 2013), and Medusagynaceae, Ochnaceae and
Quiinaceae (Matthews et al., 2012; Endress et al., 2013). However, the
variation observed in the vasculature of P. pyroidea could be related to
the establishment of the eglandular condition since, in morphs of this
species, the glands can be present or absent. Moreover, the eglandular
condition is not well defined in this species because even in the glan­
dular morph, the elaiophores can occur and be reduced in size (Cap­
pellari et al., 2011).
The populational variation of the presence and absence of elaio­
phores has already been reported for P. pyroidea (Anderson, 1997).
Further, the species’ reproductive biology indicates a new evolutionary
tendency for the group, starting with a specific mutualistic relationship
that offers oil and changing to a generalist pollination system that pro­
vides pollen (Cappellari et al., 2011). Ecologically, a hypothesis for the
presence of three petal traces would be an adaptation to new pollinating
agents. We believe that more robust petals could support the landing and
visitation of pollinators which, according to Cappellari et al. (2011), are
bees, especially bumblebees that rotate in the flower to reach all the
anthers when they are collecting pollen.
4.4. Floral structure and reproductive biology in Malpighiaceae
The literature reports several variations in the reproductive biology
of Malpighiaceae: cleistogamous flowers (Anderson, 1980b; Mamede,
1990), apomictic species (Ritzerow, 1908; Subba Rao, 1940; Singh,
1959; Lorenzo, 1981; Anderson, 1982), diversity in the
self-incompatibility system (Sigrist and Sazima, 2004; Cappellari et al.,
2011; Mendes et al., 2011) and differences in the total number of pollen
grains per anther and flower in morphs with and without elaiophores
(Teixeira and Machado, 2000). The association of the above character­
istics with changes of the plane of symmetry of the corolla indicates
changes in the reproductive system in the family, especially to a
generalist pollination system, as noted by Cappellari et al. (2011) for
P. pyroidea. Adaptation to the new reproductive system may lead to the
suppression of the glands because forming oil secretory structures is
energetically demanding for the plant, which had this characteristic due
to the reproductive success associated with oil-collecting bees.
Nothing is known about the reproductive biology of the eglandular
genera in Malpighiaceae. This is true for C. genipifolia, a species with
poricidal anthers, which may be associated with a reproductive strategy
different from all other lineages. This kind of data is necessary to un­
derstand better which agents have been exerted selective pressure on
these lineages.
4.5. Hypothetical reconstruction of the loss of elaiophores in the
acmantheroid clade
According to Puri (1951), the floral vascular ground plan comprises
the emission of three traces per sepal and one trace per petal (Fig. 7A).
The vascular ground plan modifications may include a reduction
(Fig. 7B) and fusion that, in turn, consists of connation or adnation
(Fig. 7C).
S.K.V. Bonifácio et al. Flora 281 (2021) 151870

Fig. 7. Hypotheses of possible evolutionary scenarios that culminate in eglandular calyces in Coleostachys and Pterandra pyroidea. The tree follows the most recent
phylogeny of Malpighiaceae (adapted from Davis and Anderson, 2010). A–C. Representations of floral vascular ground plan (A) and the modifications proposed by
Puri (1951). B. Reduction. C. Fusion (connation and adnation). D–F. Possible ancestral patterns of elaiophores vasculature, presented in longitudinal (left) and
transversal (right) sections. D. Elaiophore vasculature originated directly from lateral sepal traces. E. Elaiophore vasculature by acropetal traces from lateral sepal
traces of the sepal-petal complexes. F. Elaiophore vasculature by basipetal traces from lateral sepal traces of the sepal-petal complexes. G–I. Levels of synorganization
among the whorls and their relation to elaiophore vasculature. The horizontal lines with arrows indicate the levels of synorganization involving the calyx: red (sepal
traces), pink (sepal-petal complexes), and light blue (sepal-petal-antipetalous stamen complexes). G. Levels of synorganization involving calyx, corolla and
androecium. H. Hypothesis of the reduction producing an eglandular calyx; i, ii and iii indicate the respective evidence for the first hypothesis. I. Hypothesis of the
connation producing an eglandular calyx; 1, 2 and 3 indicate the respective evidence for the second hypothesis. arrowhead, the vascular bundles for the glands; one
asterisk, the byrsonimoid subclade; two asterisks, the galphimioid subclade.

Sharing vascular tissue between adjacent sepals occurs in all the
species analyzed here, which suggests that the ancestral of this clade
should already emit vascular complexes. However, this cannot be
inferred for the ancestor of the Malpighiaceae since there are no data
about the floral vasculature of Elatinaceae, the sister group of Mal­
pighiaceae (Davis and Chase, 2004). Thus, we assume that in the
vascular ground plan for Malpighiaceae, the ten elaiophores are sup­
plied by lateral sepal traces, considering they are placed marginally
between two adjacent sepals (Fig. 7D). Though, as previously demon­
strated in this work (see Figs. 2–6), acmantheroid genera exhibit
different levels of synorganization. As its consequence or as its cause, the
perianth vasculature involves connation, represented by the shared
origin of the lateral sepal traces, and adnation, represented by the
vascular complexes (Fig. 7E). Therefore, based on this evidence, prob­
ably the ancestral condition to acmantheroid clade also involves
vascular complexes formation, and glands vasculature would be sup­
plied by acropetal (Fig. 7E) and/or basipetal bundles (Fig. 7F). Very
likely, the spatial orientation of bundles depends on the gland position
and extension: in many species, the calyx glands are parallel to the se­
pals, so probably, they are vascularized by acropetal bundles (e.g.,
Janusia mediterranea; Souto and Oliveira, 2013). On the other hand, in
species with glands inserted below the sepals, which is the condition in
the studied species (Anderson et al., 2006), elaiophores are vascularized
mainly by basipetal bundles (with inverted xylem), as we registered
here. Additionally, it seems clear that all the levels of synorganization
involving sepals are directly related to elaiophores (Fig. 7G).
Considering all these points and, as well, the 10-glandular condition
of A. latifolia, we propose two different hypotheses to explain the
occurrence of eglandular calyces in P. pyroidea and C. genipifolia.
In the first hypothesis, we propose that the suppression of elaio­
phores occurs by reduction, which we observed in P. pyroidea (Fig. 7H).
Comparing the two morphs, only external morphology is changed, and
there are no significant differences between their vasculature (evidence
i); according to Puri (1951, p. 477), in some reduction cases, “[…] their
vascular stubs may persist in the tissue of the receptacle in positions
corresponding to those of the lost organs”. Moreover, comparing to
A. latifolia, it is possible to notice the numerical reduction of bundles
designated to the calyx glands (evidence ii) and the reduction of the
glands in size (evidence iii; see also Figs. 2–3). This hypothesis is
corroborated by the case of gland reduction in Tetrapterys chamaecer­
asifolia and Janusia mediterranea, species in which there are no vestigial
traces of the suppressed glands (Souto and Oliveira, 2013). The absence
of vestigial traces represents the last stage of reduction (Puri, 1951).
In the second hypothesis, we propose that connation between adja­
cent elaiophores occurred in C. genipifolia (Fig. 7I). This is mainly based
on the fusion between adjacent sepals represented by the connection of
the most external traces from branches formed by sepal-petal complexes
(evidence 1; see also Fig. 4H). Assuming that the ancestor of Coleostachys
had similar vasculature to the other genera in the clade, it is expected
that there were ten glands and two basipetal bundles per gland.

10
S.K.V. Bonifácio et al.
However, in Coleostachys, the branching that originates the basipetal
bundles is asymmetric, and one of the sepals receives a trace that divides
into two branches. In contrast, the adjacent sepal receives one trace that
gives rise to only one branch since there is a fusion between its ends.
Thus, after a fusion between elaiophores, one of the fused glands be­
comes small, and the pair disappear (evidence 2). At least, compared to
A. latifolia and considering the ancestral condition, there are four
basipetal bundles in Coleostachys; therefore, essentially, there is no
reduction in the number of traces that are emitted to the glands (evi­
dence 3). Souto and Oliveira (2013) corroborate this second hypothesis
for Mascagnia cordifolia, where traces that typically irrigate the gland of
the anterior sepal, which is eglandular in this species, are shared with
the adjacent sepal. The presence of five vestigial glands between the
sepals in some Galphimia (Anderson, 2005b, 2007), a genus of the clade
galphimioid (sister of acmantheroid), also supports this hypothesis.
5. Conclusion
Evidence shows that the presence of ten elaiophores is plesiomorphic
to Malpighiaceae, based on our observations and the literature on floral
vasculature showing the occurrence of vascular complexes and their
basipetal traces. Presently, it is not possible to affirm if the completely
eglandular calyx of Malpighiaceae is linked with connation, and in all
cases of the presence of both morphs in the same species results from
reduction. The occurrence of sepal-petal complexes in the older lineages
of Malpighiaceae points out that these structures are produced in pro­
cesses that culminate in suppressing glands. Based on this, the corolla,
commonly overlooked when discussing the numerical variations of
calyx glands in Malpighiaceae, plays a more critical role in the floral
evolution process since selective pressure by bees over 80 million years
not only involved searching for floral rewards but resulted in variation
in corolla symmetry.
Credit authorship contribution statement
Stéphani Karoline Vasconcelos Bonifácio: Conceptualization,
Methodology, Formal analysis, Validation, Investigation, Writing -
original draft, Writing - review & editing, Visualization. Rafael Felipe
de Almeida: Conceptualization, Writing - Original Draft. André Márcio
Araújo Amorim: Conceptualization, Resources, Writing - Review &
Editing, Funding acquisition. Denise Maria Trombert Oliveira:
Conceptualization, Methodology, Validation, Resources, Writing -
Original Draft, Writing - Review & Editing, Supervision, Funding
acquisition.
Declaration of Competing Interest
None.
Acknowledgments
This study was partially financed by the Coordenação de Aperfei­
çoamento de Pessoal de Nível Superior – Brazil (CAPES, Finance Code
001) and Conselho Nacional de Desenvolvimento Científico e Tec­
nológico (CNPq) – Brazil (Universal Project, process 436283/2018-2).
We thank the herbaria curators and technicians for the collection. A.
M.A. Amorim and D.M.T. Oliveira thank the CNPq for the productivity
research grants (processes 312404/2018-2 e 305686/2018-6, respec­
tively). We also acknowledge the anonymous reviewers for their valu­
able suggestions in the manuscript.
References
Almeida, R.F., Hall, C.F., 2016. Taxonomic revision of Coleostachys. Phytotaxa 277,
77–84. https://doi.org/10.1017/S0024282908007937.
Anderson, C., 1997. Revision of Pterandra (Malpighiaceae). Contrib. Univ. Michigan
Herb. 21, 1–27.
11
Flora 281 (2021) 151870
Anderson, C., 2005a. Pterandra viridiflora (Malpighiaceae), a new species from Brazil.
Contrib. Univ. Michigan Herb. 24, 21–23.
Anderson, C., 2005b. Galphimia (Malpighiaceae) in South America. Contrib. Univ.
Michigan Herb. 24, 1–12.
Anderson, C., 2007. Revision of Galphimia (Malpighiaceae). Contrib. Univ. Michigan
Herb. 25, 1–82.
Anderson, W.R., 1975. The Taxonomy of Acmanthera (Malpighiaceae). Contr. Univ.
Michigan Herb. 11, 41–50.
Anderson, W.R., 1977. Byrsonimoideae, a new sub-family of Malpighiaceae. Leandra 7,
5–18.
Anderson, W.R., 1979. Floral conservatism in Neotropical Malpighiaceae. Biotropica 11,
219–223. https://doi.org/10.2307/2388042.
Anderson, W.R., 1980a. A New Species of Acmanthera (Malpighiaceae). Syst. Bot 5,
438–441. https://doi.org/10.2307/2418524.
Anderson, W.R., 1980b. Cryptic self-fertilization in the Malpighiaceae. Science 207,
892–893. https://doi.org/10.1126/science.207.4433.892.
Anderson, W.R., 1982. Notes on Neotropical Malpighiaceae - I. Contrib. Univ. Michigan
Herb. 15, 93–136.
Anderson, W.R., 1990a. The Origin of the Malpighiaceae - the evidence from
morphology. Mem. N. Y. Bot. Gard. 64, 210–224.
Anderson, W.R., 1990b. Notes on Neotropical Malpighiaceae. – III. Contr. Univ. Michigan
Herb. 17, 39–54.
Anderson, W.R., Anderson, C., Davis, C.C., 2006. Malpighiaceae http://herbarium.lsa.
umich.edu/malpigh/index.html (accessed 23.11.21).
Cappellari, S.C., Haleem, M.A., Marsaioli, A.J., Tidon, R., Simpson, B.B., 2011. Pterandra
pyroidea: a case of pollination shift within Neotropical Malpighiaceae. Ann. Bot. 107,
1323–1334. https://doi.org/10.1093/aob/mcr084.
Carvalho, P.D., Borba, E.L., Lucchese, A.M., 2005. Variação no número de glândulas e
produção de óleo em flores de Stigmaphyllon paralias A. Juss. (Malpighiaceae). Acta
Bot. Bras. 19, 209–214. https://doi.org/10.1590/S0102-33062005000200002.
Davis, C.C., Chase, M.W., 2004. Elatinaceae are sister to Malpighiaceae; Peridiscaceae
belong to Saxifragales. Am. J. Botany 91, 262–273. https://doi.org/10.3732/
ajb.91.2.262.
Davis, C.C., Anderson, W.R., 2010. A complete generic phylogeny of Malpighiaceae
inferred from nucleotide sequence data and morphology. Am. J. Bot. 97, 2031–2048.
https://doi.org/10.3732/ajb.1000146.
Davis, C.C., Anderson, W.R., Donoghue, M.J., 2001. Phylogeny of Malpighiaceae:
evidence from chloroplast ndhF and trnL-F nucleotide sequences. Am. J. Bot. 88,
1830–1846.
Davis, C.C., Schaefer, H., Xi, Z., Baum, D.A., Donoghue, M.J., Harmon, L.J., 2014. Long-
term morphological stasis maintained by a plant-pollinator mutualism. Proc. Natl.
Acad. Sci. U. S. A. 111, 5914–5919. https://doi.org/10.1073/pnas.1403157111.
Davis, C.C., Marinho, L.C., Amorim, A.M.A., 2020a. Andersoniella: a new genus of
neotropical Malpighiaceae. Harvard Pap. Bot. 25, 51–56. https://doi.org/10.3100/
hpib.v25iss1.2020.n6.
Davis, C.C., Marinho, L.C., Amorim, A.M.A., 2020b. Andersoniodoxa, a replacement name
for Andersoniella (Malpighiaceae). Phytotaxa 470, 121–122. https://doi.org/
10.11646/phytotaxa.470.1.9.
Eames, A.J., MacDaniels, L.H., 1947. An introduction to plant anatomy, 2nd ed. Mc Graw
Hill, New York.
Endress, P.K., Davis, C.C., Matthews, M.L., 2013. Advances in the floral structural
characterization of the major subclades of Malpighiales, one of the largest orders of
flowering plants. Ann. Bot. 111, 969–985. https://doi.org/10.1093/aob/mct056.
Farroñay, F., Oliveira-Perdiz, R., Prata, E.M.B., Vicentini, A., 2019. Notes on morphology
and distribution of Acmanthera (A. Juss.) Griseb. (Malpighiaceae), an endemic genus
from Brazil. Phytotaxa 415, 199–207. https://doi.org/10.11646/phytotaxa.415.4.4.
Gifford, E.M., Foster, A.S., 1989. Morphology and evolution of vascular plants, 3rd ed.
W.H. Freeman and Company, San Francisco.
Gonçalves-Souza, P., Schlindwein, C., Paiva, E.A.S., 2018. Floral resins of Philodendron
adamantinum (Araceae): Secretion, release and synchrony with pollinators. Acta Bot.
Bras. 32, 392–401. https://doi.org/10.1590/0102-33062018abb0115.
Gong, Y.B., Huang, S.Q., 2009. Floral symmetry: Pollinator-mediated stabilizing
selection on flower size in bilateral species. Proc. R. Soc. B Biol. Sci. 276, 4013–4020.
https://doi.org/10.1098/rspb.2009.1254.
Guesdon, I.R., Amorim, A.M., Meira, R.M.S.A., 2019. Functional role and evolutionary
contributions of floral gland morphoanatomy in the Paleotropical genus Acridocarpus
(Malpighiaceae). PLoS One 14, 1–20. https://doi.org/10.1371/journal.
pone.0222561.
Lorenzo, E., 1981. Sobre la inflorescencia, morfologia floral y embriologia de Janusia
guaranitica (Malpighiaceae). Kurtziana 14, 101–124.
Mamede, M.C.H., 1990. Revisão do gênero Camarea Saint-Hilaire (Malpighiaceae).
Hoehnea 17, 1–34.
Matthews, M.L., Endress, P.K., 2011. Comparative floral structure and systematics in
Rhizophoraceae, Erythroxylaceae and the potentially related Ctenolophonaceae,
Linaceae, Irvingiaceae and Caryocaraceae (Malpighiales). Bot. J. Linn. Soc. 166,
331–416. https://doi.org/10.1111/j.1095-8339.2011.01162.x.
Matthews, M.L., Amaral, M.D.C.E., Endress, P.K., 2012. Comparative floral structure and
systematics in Ochnaceae s.l. (Ochnaceae, Quiinaceae and Medusagynaceae;
Malpighiales). Bot. J. Linn. Soc. 170, 299–392. https://doi.org/10.1111/j.1095-
8339.2012.01299.x.
Mello, A.C.M.P., De Almeida, R.F., Amorim, A.M.A., Oliveira, D.M.T., 2019. Leaf
structure in Amorimia and closely related Neotropical genera and implications for
their systematics and leaf evolution in Malpighiaceae. Bot. J. Linn. Soc. 191,
102–127. https://doi.org/10.1093/botlinnean/boz028.
Mendes, F.N., Rêgo, M.M.C., de Albuquerque, P.M.C., 2011. Fenologia e biologia
reprodutiva de duas espécies de Byrsonima Rich. (Malpighiaceae) em área de Cerrado
S.K.V. Bonifácio et al.
no Nordeste do Brasil. Biota Neotrop 11, 103–115. https://doi.org/10.1590/S1676-
06032011000400011.
O’Brien, T.P., Feder, N., McCully, M.E., 1964. Polychromatic staining of plant cell walls
by toluidine blue O. Protoplasma 59, 368–373. https://doi.org/10.1007/
BF01248568.
Paiva, E.A.S., Pinho, S.Z., Oliveira, D.M.T., 2011. Large plant samples: how to process for
GMA embedding? In: Chiarini-Garcia, H., Melo, R.C.N. (Eds.), Light microscopy:
methods and protocols. Springer, New York, pp. 37–49. https://doi.org/10.1007/
978-1-60761-950-5.
Puri, V., 1951. The role of floral anatomy in the solution of morphological problems. Bot.
Rev. 17, 471–553. https://doi.org/10.1007/BF02882536.
Renner, S.S., Schaefer, H., 2010. The evolution and loss of oil-offering flowers: New
insights from dated phylogenies for angiosperms and bees. Philos. Trans. R. Soc. B
Biol. Sci. 365, 423–435. https://doi.org/10.1098/rstb.2009.0229.
Ritzerow, H., 1908. Über Bau und Befruchtung kleistogamer Blüten. Flora 98, 163–212.
Sazima, M., Sazima, I., 1989. Oil-gathering bees visit flowers of eglandular morphs of the
oil-producing Malpighiaceae. Bot. Acta 102, 106–111. https://doi.org/10.1111/
j.1438-8677.1989.tb00073.x.
Sigrist, M.R., Sazima, M., 2004. Pollination and reproductive biology of twelve species of
neotropical Malpighiaceae: stigma morphology and its implications for the breeding
system. Ann. Bot. 94, 33–41. https://doi.org/10.1093/aob/mch108.
12
Flora 281 (2021) 151870
Simpson, B.B., Neff, J.L., 1981. Floral rewards: alternatives to pollen and nectar, 68. Ann.
Missouri Bot. Gard, pp. 301–322. https://doi.org/10.2307/2398800.
Singh, B., 1959. Studies in the family Malpighiaceae: I. Morphology of Thryallis glauca
Kuntze. Hortic. Adv. 3, 1–19.
Smith, F.H., Smith, E.C., 1942. Anatomy of the Inferior Ovary of Darbya. Am. J. Bot. 29,
464. https://doi.org/10.2307/2437312.
Souto, L.S., Oliveira, D.M.T., 2013. Evaluation of the floral vasculature of the Janusia,
Mascagnia and Tetrapterys species as a tool to explain the decrease of floral organs in
Malpighiaceae. Flora 208, 351–359. https://doi.org/10.1016/j.flora.2013.05.002.
Subba Rao, A.M., 1940. Studies in the Malpighiaceae - I Embryo sac development and
embryogeny in the genera Hiptage, Banisteria and Stigmatophyllum. J. Indian Bot. Soc.
18, 145–156.
Teixeira, L.A.G., Machado, I.C., 2000. Sistema de polinização e reprodução de Byrsonima
sericea DC (Malpighiaceae). Acta Bot. Bras. 14, 347–357. https://doi.org/10.1590/
S0102-33062000000300011.
Vogel, S., 1974. Ölblumen und ölsammelnde Bienen. Trop. und Subtrop. Pflanzenwelt 7,
283–547.
Vogel, S., 1990. History of the Malpighiaceae pollination ecology. Mem. N. Y. Bot. Gard.
55, 130–142.
Zhang, W., Kramer, E.M., Davis, C.C., 2010. Floral symmetry genes and the origin and
maintenance of zygomorphy in a plant-pollinator mutualism. Proc. Natl. Acad. Sci.
U. S. A. 107, 6388–6393. https://doi.org/10.1073/pnas.0910155107.