Int. J. Plant Sci. 175(7):814–827. 2014.

䉷 2014 by The University of Chicago. All rights reserved.

1058-5893/2014/17507-0008$15.00 DOI: 10.1086/676817

COMPARATIVE ANATOMY OF REPRODUCTIVE STRUCTURES

IN CYCLANTHACEAE (PANDANALES)
M. Graça Sajo,* Julio A. Lombardi,* Rafaela C. Forzza,† and Paula J. Rudall1,‡
*Instituto de Biociências, Universidade Estadual Paulista (UNESP), Rio Claro, CEP 13506-900, SP, Brazil; †Jardim Botânico do Rio de Janeiro,
CEP 22470-180, RJ, Brazil; and ‡Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3NJ, United Kingdom

Editor: Alan W. Meerow

Premise of research. The Neotropical family Cyclanthaceae is one of five families of the order Pandanales,
which includes some misfit taxa that are morphologically highly anomalous. In the taxonomically isolated
genus Cyclanthus, which is sister to all other Cyclanthaceae, individual flowers are not readily distinguishable.
Instead, flowers are replaced by discoidal rings that encircle the inflorescence. We compare features that are
shared among misfit taxa.
Methodology. We use scanning electron microscopy and light microscopy to describe the reproductive
structures of both subfamilies of Cyclanthaceae, including Cyclanthus bipartitus (Cyclanthoideae) and several
Carludovicoideae.
Pivotal results. Our pollination observations on Carludovica palmata flowers contrast with some other
reports, because we observed only small black bees of the Meliponini tribe visiting this species. The anomalous
reproductive structures of Cyclanthus show some features in common with Carludovicoideae, especially the
presence of large axial mucilage canals. However, Cyclanthus differs from Carludovicoideae in several respects;
for example, the outer integument is relatively poorly developed in Cyclanthus. The staminodes on male
flowers of Carludovicoideae are secretory, with elongated epidermal cells.
Conclusions. We interpret as staminodes the sterile secretory lobes that are present on the androphore
of most Carludovicoideae. Our observations enhance existing knowledge of floral anatomy of Cyclanthaceae,
especially Cyclanthus, and provide a comparative basis for future studies of Pandanales.

Keywords: anatomy, Carludovicoideae, Cyclanthus, pseudanthia, staminodes, tapetum.

Introduction 1998). Little detailed comparative anatomical information is

currently available on the unusual reproductive structures of
The order Pandanales, with five families, includes several this family, especially with respect to Cyclanthus. We inves-
misfit taxa that possess some highly anomalous morphological tigate the anatomy of the reproductive units of several species
features (Rudall and Bateman 2006). Perhaps the most notable of Cyclanthaceae, including one species of Cyclanthus and
examples are the inside-out flowers of Lacandonia (Triurida- seven (of 11) genera of Carludovicoideae (table 1). Our pri-
ceae), the pentamerous flowers of Pentastemona (Stemona- mary aims are to improve our understanding of the organi-
ceae), the baseball-like floral units of Sararanga (Pandanaceae), zation, homologies, and functional biology of these unusual
and the discoidal rings that replace groups of flowers on the structures. Our ultimate goal is to compare features that are
inflorescence of Cyclanthus (Cyclanthaceae). The homologies shared between misfit taxa of the anomalous order Pandanales
of some of these structures remain unresolved, partly because with respect to the apparent loss of developmental constraints
of conflicting data but also partly because scant material is that occurred during the evolution of the order.
available for comparative studies. Contrasting modes of gender
distribution characterize different families of Pandanales; for Background: Systematics and Morphology
example, Pandanaceae are dioecious, and Cyclanthaceae are
A close relationship between Cyclanthaceae and the paleo-
monoecious.
tropical family Pandanaceae was previously indicated by
In this article, we focus on Cyclanthaceae (12 genera, ca.
strong similarities in morphology (e.g., Harling 1958; Dahl-
230 species; table 1), a monocot family of perennial herbs,
gren et al. 1985) and is well supported by molecular data (e.g.,
shrubs, vines, and epiphytes that grow in Neotropical wet,
Graham et al. 2006). Harling (1958) subdivided Cyclanthaceae
shaded forests in Central and South America (Harling et al.
into two somewhat unbalanced subfamilies (table 1), Carlu-
dovicoideae (11 genera, ca. 230 species) and Cyclanthoideae,
1
Author for correspondence; e-mail: [email protected]. which includes a single genus with a single extant species,
Manuscript received February 2014; revised manuscript received March 2014; Cyclanthus bipartitus, and an extinct species, Cyclanthus mes-
electronically published August 8, 2014. selensis (Smith et al. 2008). Cyclanthus bipartitus has been

814
 SAJO ET AL.—FLOWERS OF CYCLANTHACEAE 815

Table 1
Genera of Cyclanthaceae and Material Examined
Genus No. species Material examined
Subfamily Cyclanthoideae:
Cyclanthus Poit. 1 Cyclanthus bipartitus Poit. ex A.Rich. (RC: Sajo s.n.; SEM/LM)
Subfamily Carludovicoideae:
Asplundia Harling Ca. 100 Asplundia humilis (Poepp. and Endl.) Harling (M: Bogner s.n.), As-
plundia aff. maximiliani Harling (BHCB: Lombardi 5408; HRCB:
Lombardi 7636; SEM/LM)
Carludovica Ruiz and Pav. 4 Carludovica palmata Ruiz and Pav. (RC: Sajo s.n.; HK 1984-2120)
Chorigyne R.Erikss. 7 Chorigyne pendula (Hammel) R.Erikss. (M: Bogner s.n.)
Dicranopygium Harling Ca. 50 Dicranopygium atrovirens (H.Wendl.) Harling (M: Bognor s.n.), Di-
cranopygium microcephalum (Hook.f.) Harling (M: Bogner s.n.),
Dicranopygium pygmaeum (Gleason) Harling (M: Bogner s.n.)
Dianthoveus Hammel and Wilder 1 ...
Evodianthus Oerst. 1 Evodianthus funifer (Poit.) Lindm. (HRCB: Lombardi 8788)
Ludovia Brongn. 3 Ludovia lancifolia Brongn. (M: Bogner s.n.; SEM/LM)
Schultesiophytum Harling 1 ...
Sphaeradenia Harling Ca. 50 Sphaeradenia woodsonii Harling (M: Bogner s.n.)
Stelestylis Drude 4 ...
Thoracocarpus Harling 1 ...
Note. s.n. p no number available.

placed as sister to the rest of the family in all phylogenetic found that some of these structures appear more discrete (and
analyses to date, using both morphological data (Eriksson therefore more flowerlike) at very early developmental stages,
1994a) and molecular data (Davis et al. 2004; Mennes et al. though true flowers are difficult to distinguish. Smith et al.
2013), though molecular analyses to date have not included (2008) found remarkably similar fruiting cycles in a fossil from
all genera of Carludovicoideae. the Middle Eocene of Messel, Germany, which they described
Leaves of Cyclanthaceae are characteristically petiolate, pli- as a new, extinct species of Cyclanthus (C. messelensis); this
cate, and often deeply divided. The interesting and complex species grew in the Paleogene of the Old World, indicating a
pollination biology of Cyclanthaceae, involving weevil life cy- more widespread ancient distribution for the family.
cles, has been studied in several species (Beach 1982; Eriksson
1993, 1994b; Franz 2004). Inflorescences are typically un-
branched, spadix-like, enclosed by several spathe-like bracts
before anthesis, and monoecious, bearing both staminate and Material and Methods
pistillate unisexual flowers in characteristic arrangements.
Flowers of subfamily Carludovicoideae are sessile and borne Material examined is listed in table 1. Some material was
in clusters on a central rachis, with each cluster consisting of collected from plants in their natural habitats, for which
a single central pistillate flower surrounded by approximately vouchers were deposited in the Brazilian herbaria, listed in
four staminate flowers (Harling 1958; Harling et al. 1998). In table 1 as HRCB and BHCB, followed by the collector’s name
some species of Carludovicoideae, the individual flowers are and collection number. Other material was collected from cul-
free from each other, and in others they are fused together, tivated plants grown at the Royal Botanic Gardens, Kew (HK);
though each flower remains identifiable. Each flower cluster Munich Botanic Garden (M; donated by Josef Bogner); or a
represents a partial inflorescence with a terminal pistillate private collection in Rio Claro, SP, Brazil (RC). Observations
flower (Harling 1958). Remarkably, each partial inflorescence on pollination in Carludovica palmata were made in early
is functionally equivalent to a flower, with a central pistillate January 2010 and 2013 by one of us (M. G. Sajo) using cul-
structure surrounded by staminate structures (Eriksson tivated plants growing in Parati (RJ), Brazil.
1994a). In some Carludovicoideae (e.g., Thoracocarpus bis- Some Cyclanthaceae, especially C. palmata and Cyclanthus
sectus), each partial inflorescence is subtended by an incon- bipartitus, are widely grown as ornamentals for their hand-
spicuous bract at early developmental stages, though these some leaves. Inflorescences at various stages of development
bracts become undetectable in surface view soon after their were available for some Carludovicoideae, but very early
initiation (Wilder 1977). stages are difficult to obtain without destroying plants. When
In contrast, in Cyclanthoideae (Cyclanthus), the stamens and inflorescences are visible, their floral organs are already well
carpels are inserted on the primary inflorescence axis in char- developed.
acteristic rings, or cycles, which differ from the flowers of all Inflorescences were fixed in formalin–acetic acid–alcohol
other angiosperms, making individual flowers impossible to and stored in 70% ethanol. For light microscopy, pieces of
differentiate at maturity and leading some authors to describe each inflorescence were embedded in Paraplast using standard
them as pseudanthia (Dahlgren et al. 1985; Rudall and Bate- methods and serially sectioned at ca. 12-mm thickness using a
man 2006; Endress 2014). In a detailed study, Wilder (1981) rotary microtome. For light microscopy (LM), sections were
816 INTERNATIONAL JOURNAL OF PLANT SCIENCES

Fig. 1 Carludovica palmata, reproductive sequence. A, Early-morning inflorescence, just opening. B, Inflorescence at 9:30 a.m., the outer
spathe fallen. C, The remaining spathes unfolded, exposing the staminodes, which attract small black bees. D, Staminodes starting to elongate;
more bees walk among the staminodes (apparently eating them) and reach the tops of the staminate and pistillate flowers. E, F, By the following
morning, the staminodes are abscised, and sticky drops are visible in the top of the pistillate flowers of some inflorescences. G, Subsequently,
the staminate units start to abscise; bees now collect pollen. H, Some inflorescences develop red fleshy baccate fruits. J, A single visiting bee of
the Meliponini tribe.

mounted onto microscope slides, stained in safranin and Alcian Zeiss Axiocam digital camera. For scanning electron micro-
blue, dehydrated through an ethanol series to 100% ethanol, scope (SEM) examination, fixed inflorescences were carefully
transferred to Histoclear, and mounted in DPX mounting me- dissected in 70% ethanol and then dehydrated in an ethanol
dium (distrene, with dibutyl phthalate and xylene). Slides were series to 100% ethanol. Then they were critical-point dried
examined using a Leica DMLB photomicroscope fitted with a using a Bal-Tec 030 critical-point dryer, mounted onto pin
 SAJO ET AL.—FLOWERS OF CYCLANTHACEAE 817

the tops of the staminate and pistillate flowers. We did not

observe any beetle visiting the flowers or any insect visitors in
the late evening (up to midnight). Staminode elongation is
complete by the early evening of the same day (ca. 6:30 p.m.),
and the following morning the staminodes all abcise (fig. 1E).
At this stage, secretory drops are visible at the top of the
pistillate flowers, at least in some inflorescences (fig. 1F). Sub-
sequently, the staminate flowers start to abscise (fig. 1G), but
the inflorescence is still visited by individuals of the same bee
species, which now collect pollen (fig. 1J). After all the sta-
minate flowers have abscised, some inflorescences develop into
multiple red fleshy baccate fruits (fig. 1H), while others
degenerate.
Reproductive structures. In all Carludovicoideae, 3–5 sta-
minate flowers surround a single pistillate flower (figs. 2, 3).
Each staminate flower consists of many stamens with basifixed
anthers. The stamens are fused together at their bases, forming
a triangular stalk, or “androphore,” a term that is used var-
iously to describe either an extension of the receptacle or a
column formed by united filaments. In most Carludovicoideae
(except Evodianthus funifer), the androphore bears small ster-
ile lobes surrounding the free stamens; these lobes are located
mainly in the region away from the pistillate flower (figs. 3C,
3G, 3J, 3K, 4F). Each sterile lobe lacks vasculature and consists
of a single-layered epidermis enclosing 8–10 layers of paren-
chyma cells; the abaxial epidermal cells are anticlinally elon-
gated and appear secretory (fig. 4G). Mature stamen filaments
often contain tannins (fig. 4I, 4J).
Each stamen is supplied by a single collateral vascular bun-
dle (fig. 4E). Within the androphore, the individual stamen
bundles are mostly distinct from each other but are fused into
a central vascular plexus at the androphore base (fig. 4I). The
anther wall (fig. 4H) consists of a papillate outer epidermis;
Fig. 2 Carludovicoideae, photographs of inflorescences (fixed ma- an endothecial layer of anticlinally elongated, thick-walled
terial). A, Ludovia lancifolia, base of dissected unopened inflorescence cells; a second middle layer that is thin walled and ephemeral;
with bracts removed. B, Evodianthus funifer, older, postanthetic in- and an inner tapetum, indicating the monocot type of anther
florescence. C, Sphaeradenia woodsonii, dissected unopened inflores- wall development in Carludovicoideae, as in other Pandanales.
cence with bracts removed. D, Dicranopygium microcephalium, young The tapetum is probably secretory, though its cells are at least
inflorescence with bracts partially removed. partially dispersed into the anther locule at the tetrad stage
(fig. 4H). The anther dehisces by longitudinal slits, and the
stubs, coated with platinum using an Emitech K550 sputter- endothecium is persistent after anthesis (fig. 4K). A persistent
coater, and examined using a Hitachi cold-field emission SEM crystal idioblast is present in the hinge of each anther locule
S-4700 at 2 kV. (fig. 4K), ultimately dispersing raphides into the locule.
In all Carludovicoideae, each pistillate flower possesses an
Results outer whorl of 4–5 thick perianth lobes (fig. 3A, 3E, 3F), which
remain poorly developed in Ludovia. The number of perianth
lobes varies within an inflorescence. Each perianth lobe has a
Carludovicoideae
parenchymatous mesophyll that contains many collateral vas-
Pollination and developmental observations for Carludovica cular bundles; sometimes groups of thick-walled cells are also
palmate. Many inflorescences develop in sequence during present. Within the perianth whorl, a corresponding whorl of
the reproductive season, which coincides with spring-summer 3–5 staminodes surrounds an alternating whorl of 3–5 carpels.
in São Paulo (mid-October to early January). Typically, several Each staminode is fused at its base to the corresponding peri-
inflorescences at different stages are present at the same time anth lobe. Each staminode possesses a single collateral bundle
on the same plant. Each inflorescence is first visible in early that diverges from the central bundle of the corresponding
morning (fig. 1A). At ca. 9:30 a.m., the outer inflorescence perianth lobe. The carpels are equal in number to the stami-
spathe falls off (fig. 1B), and the remaining spathes unfold and nodes and alternate with them (fig. 5B). All the organs are
expose the staminodes (fig. 1C). An hour later, the staminodes united at their bases, and the carpels are partially embedded
start to elongate (fig. 1D). The inflorescence is then visited by in the inflorescence axis (fig. 5A). There is a single ovary locule
small black bees (fig. 1C, 1D) that travel among the staminodes (figs. 5C, 5F, 6F). Each carpel is supplied by numerous col-
(apparently eating them) and also move under them, reaching lateral vascular bundles; the supranumerary carpel bundles
 Fig. 3 Carludovicoideae, SEMs. A, Ludovia lancifolia, pistillate flower (right) and one of four staminate flowers (left). B, Ludovia lancifolia,
detail of sterile lobes on outside edge of staminate flower. C, Asplundia sp., staminate flower. D, Asplundia sp., pistillate flower (at center) with
surrounding staminate flowers (visible at top). E, Asplundia brachyphylla. F, Dicranopygium atrovirens, pistillate flower (right) and one of three
staminate flowers (left). G, Carludovica palmata, staminate flower. H, Dicranopygium microcephalium, tips of staminodes of pistillate flowers.
J, Dicranopygium atrovirens, staminate flower from below. K, Dicranopygium atrovirens, two staminate flowers, with part of pistillate flower
on right. and p androphore, sl p sterile lobe, stde p staminode, stig p stigma. Scale bars p 1 mm in A, C–F; 100 mm in B, H; 500 mm in
G, J, K.
 SAJO ET AL.—FLOWERS OF CYCLANTHACEAE 819

Fig. 4 Carludovicoideae, staminate flowers (LM). A–H, Asplundia sp., preanthetic male flowers (B–E, very young flower; A, F, G, older
flowers). A, Median longitudinal section (LS) through the androphores of two male flowers. B, C, Serial transverse sections (TS) through anthers
(B) and filaments/androphore (C) of a single very young male flower. D, TS young preanthetic anther. E, TS filament, showing single vascular
bundle. F, TS through edge of androphore, showing sterile lobes. G, TS sterile lobe, showing secretory outer epidermis. H, TS anther, showing
wall layers. I–K, Carludovica palmata. I, TS base of androphore, showing vascular plexus. J, TS filaments with dark-staining tannin cells. K,
TS dehisced anthers. an p androphore, ci p crystal idioblast, end p endothecial layer, ml p middle layer, oe p papillate outer epidermis of
anther wall, pt p pollen tetrads, sl p sterile lobe, tap p tapetum, vb p vascular bundle, vp p vascular plexus. Scale bars p 100 mm in A–
F, I–K; 50 mm in G, H.

make it difficult to distinguish the dorsal and lateral bundles. dles are those located in the center of the placentae, which
Carpels of the majority of angiosperms typically have three supply the ovules (fig. 5C). A stigmatic region is present at the
bundles: two lateral bundles that supply the ovules and a dorsal top of each carpel (fig. 5A, 5D).
bundle that supplies the stigma. Thus, in our material, the Mucilage canals are absent during early development but per-
dorsal carpellary bundles are probably those located between meate the tissues of older flowers (fig. 5D). Crystal idioblasts
the placentae at the ovary base, and the lateral carpellary bun- are widespread in parenchymatous tissues of all species; they
820 INTERNATIONAL JOURNAL OF PLANT SCIENCES

Fig. 5 Carludovicoideae, pistillate flowers (LM). A–C, Asplundia sp., preanthetic flowers. A, Longitudinal section (LS); ovary is unilocular
but sectioned through placenta. B, Transverse section (TS) through staminodes and upper (free) parts of carpels. C, TS of symplicate zone of
ovary, showing single locule and five separate placentas bearing young ovules. D, Carludovica palmata, LS older flower with mucilage canals;
ovary is unilocular but sectioned through placenta. cm p carpel margin, dcb p dorsal carpellary bundle, mc p mucilage canal, ovl p ovary
locule with ovules, plac p placenta, po p perianth organ, scl p stylar canal, stde p staminode, stfl p staminate flower, stig p stigma, vcb p
lateral carpellary bundle. Scale bars p 100 mm.

consist of a bundle of raphide crystals embedded in mucilage. partially fused with the raphe region; the micropyle is formed
As the flower ages, the idioblasts enlarge and coalesce with each by the inner integument alone. The megasporocyte develops
other and with other parenchyma cells to form mucilage canals. into a female gametophyte of the Polygonum type. At the me-
The mucilage canals are usually axially (longitudinally) oriented, gasporocyte stage, the nucellus consists of a reduced hypostase
especially in regions surrounding the ovary locules. and a layer of anticlinally elongated dermal cells at the mi-
The ovules (fig. 6B–6E) are anatropous, and their curvature cropylar end. The outer integument is three layered and mul-
commences just after the integuments are initiated. Each ovule tilayered at the micropyle; the inner integument is two layered
possesses two integuments, though the outer integument is for most of its length and multilayered at the micropyle.
 SAJO ET AL.—FLOWERS OF CYCLANTHACEAE 821

Fig. 6 Carludovicoideae (LM). A, Asplundia brachyphylla, transverse section (TS) ovary locule with ovules. B, Asplundia sp., early ovule
at megasporocyte stage. C, Carludovica palmata, mature ovule. D, E, Ludovia lancifolia, mature ovules. F, Asplundia brachyphylla, TS staminode,
showing single vascular bundle. es p embryo sac, ovl p ovary locule with ovules, plac p placenta, stde p staminode. Scale bars p 100 mm
in A, D–F; 20 mm in B; 50 mm in C.

Cyclanthoideae induce flowering. We obtained a living inflorescence of this

species for detailed anatomical investigation (figs. 7–10). Sta-
Cyclanthus bipartitus flowers rarely, either in cultivation or
minate and pistillate structures are arranged in alternating
in the wild, though Wilder (1981) examined a range of de-
velopmental stages. Beach (1982) reported that C. bipartitus rings or cycles throughout most of the spadix. In our material
is fairly abundant throughout the primary and secondary for- there was a group of stamens at the tip (fig. 7).
ests near the La Selva Field Station in Costa Rica, where it The staminate cycles consist entirely of stamens inserted in
flowers in April and May, but fewer than 5% of the mature 3–4 rows, with their filaments mostly free from each other but
plants produced flowers during the period of his study. In often laterally fused at their bases (fig. 9E). Each stamen is
cultivation, spraying with ethylene in the greenhouse failed to supplied by a single collateral bundle (fig. 10C). In contrast
822 INTERNATIONAL JOURNAL OF PLANT SCIENCES

collateral bundles approaches the bases of the perianth lobes

without entering them. A single vascular bundle (a fusion prod-
uct of the lateral and dorsal carpellary bundles) enters each
stigma.
Many ovules are present in the ovary canals of the pistillate
rings (figs. 9, 10). The ovules range from campylotropous to
anatropous and are inserted in a parietal placenta. The ovules
are each covered by a single inner integument that is three
layered; the outer integument is relatively short. The nucellus
possesses a relatively reduced hypostase and an epidermis of
anticlinally elongated cells at the micropylar end.

Discussion

Functional Biology
The pollination biology of Cyclanthus is relatively poorly
known. Beach (1982) reported that the inner pair of inflores-
cence bracts differs from the outer pair in that their adaxial
Fig. 7 Cyclanthus bipartitus (Cyclanthoideae), different views of
surfaces consist of a specialized mealy tissue that provides a
the same very young inflorescence with outer bracts removed. A, En-
lipid-rich food source for pollinating beetles that remain on
tire. B, Median longitudinal section. C, Transverse section. D, Entire.
E, Detail. ov p ovary locule, p p pistillate cycle, s p staminate cycle. the spadix for long periods.
Some Carludovicoideae have been studied in more detail. In
general, our field observations of Carludovica palmata flowers
with Carludovicoideae, the anthers are basifixed (fig. 8C), and
agree with those of Schremmer (1982) but contrast with other
the anther walls consist of four cell layers (fig. 10C): a papillate
reports. Several authors (Harling 1958; Gottsberger 1990,
epidermis, an indistinct tapetum, and two intervening layers
1991; Eriksson 1994b) have reported that the inflorescences
with a relatively undifferentiated endothecium. At stages ex-
of all Carludovicoideae have similar functional morphology
amined here, the anther locule was filled with cytoplasmic
and floral phenology, with weevils (Curculionidae) acting as
material (fig. 10C).
pollinators. Gottsberger (1990, 1991) observed beetles visiting
Each pistillate cycle includes three distinct structures: a ring
the inflorescences at the stage when the staminodes are elon-
of carpels (each bearing stigmatic lobes), a ring of sterile stam-
gating; the weevils copulate on the inflorescences and facilitate
inodes on either side of the carpels, and a ring of sterile peri-
pollination. However, like Schremmer (1982), we observed
anth lobes on either side of each staminode ring (figs. 8C, 9,
only small black bees of the Meliponini tribe visiting Carlu-
10B). All these structures are distally free from each other but
dovica; the bees reach the region where the staminate and
proximally united (for most of their length) within each pis-
pistillate reproductive units are located (figs. 3, 4). We observed
tillate cycle. The staminodes each possess a distal, narrow con-
no beetles visiting the flowers, during either the day or late
striction below the sterile anther (figs. 8B, 9A). Each staminode
evenings (up to midnight), though we cannot exclude the pos-
is supplied by a single collateral bundle that divides in the
sibility that beetles visited at night or in the very early morning.
pseudoanther region (fig. 10B). The perianth lobes are not
Some authors refer to a scent emitted by the staminodes to
showy and are typically obscured by the overarching stami-
attract the pollinators (Drude 1877; Knuth 1904; Harling
nodes (figs. 8C, 9A); in their free apical regions they are
1958; Schremmer 1982; Gottsberger 1990, 1991; Eriksson
unvascularized.
1994b). Our observations show that secretory cells cover the
The base of the ovary locule is slightly sunken into the in-
abaxial surfaces of the sterile lobes of the staminate units. This
florescence axis (figs. 8C, 9A). Mucilage canals extend axially
secretion could participate in attraction of pollinators and
(longitudinally) throughout each pistillate unit (fig. 9). Close
other visitors, though there are conflicting reports of the timing
to the spadix, the staminate and pistillate cycles are united,
of the different reproductive stages. Some floral biologists con-
and the ovaries form a single ovary locule that is continuous
sider the inflorescence of Carludovicoideae to be protogynous,
around the entire axis (fig. 9). At more distal cross-sectional
with the female phase lasting about 2 d, followed by the open-
levels (away from the inflorescence axis), the staminate cycles
ing of the anthers (Drude 1877; Knuth 1904; Harling 1958;
become identifiable first by the strands of collateral bundles
Schremmer 1982; Gottsberger 1990, 1991; Eriksson 1994b).
that irrigate the stamens and then by the individual filaments
However, in common with those of Schremmer (1982), our
(fig. 9). At these levels, 3–4 rows of collateral bundles are
observations reject protogyny in Carludovicoideae. We ob-
located on each side of the ovary canal. The bundles adjacent
served anther dehiscence on the second day in C. palmata, just
to the mucilage canals are the dorsal carpellary bundles (figs.
after the staminodes are shed, when the ovules are at the mega-
9, 10A). Above the ovary canal, the dorsal carpellary bundles
gametophyte phase.
alternate with collateral bundles that supply the staminodes
(fig. 9B). At the same level, 2–3 lateral bundles per carpel are
Family Traits
distinguishable; the lateral bundles supply the ovules. Close to
the top of the pistillate unit, the lateral bundles of each carpel The anomalous genus Cyclanthus shares some distinctive
fuse with the corresponding dorsal bundle. An outer row of features with other Cyclanthaceae, especially the presence of
 SAJO ET AL.—FLOWERS OF CYCLANTHACEAE 823

Fig. 8 Cyclanthus bipartitus (Cyclanthoideae; SEM). A, Sterile tips of three staminodes. B, Surface view of staminodes and stigmatic lobes.
C, Longitudinal section through adjacent staminate and pistillate cycles. an p anther, ov p ovules, pl p perianth lobe, stde p staminode,
stig p stigma. Scale bars p 500 mm in A; 1 mm in B, C.

large axial mucilage canals that initially develop by fusion of for example, Carludovicoideae and Stemonaceae all possess
numerous large raphide idioblasts and become more promi- secretory tapeta with orbicules (Furness and Rudall 1998,
nent and extensive with age. These canals could be related to 2006; Harling et al. 1998; Sajo et al. 2013; this article). How-
the functional biology of the reproductive structures of Cy- ever, the plasmodial type occurs in some Pandanaceae (e.g., in
clanthaceae, though their presence close to the ovular region Freycinetia; Furness and Rudall 2001). An intermediate (in-
indicates that they form a physical barrier to predation of the vasive-nonsyncytial) type occurs in some Triuridaceae and Vel-
ovules and seeds by insects. Similar mucilage canals are not loziaceae, in which the tapetal cell walls degenerate at the
reported in other Pandanales, except in some Triuridaceae tetrad stage and the multinucleate tapetal protoplasts disso-
(Rudall 2003). ciate into the anther locule but do not form a true plasmodium.
The supranumerary carpel bundles that we report here in Furness and Rudall (2006) speculated that this intermediate
Carludovicoideae (though not in Cyclanthus) are probably re- type of tapetum could be relatively common in Pandanales,
lated partly to the relatively large carpel size and partly to the which could explain the numerous anomalous and conflicting
wall complexity of the developing fleshy fruit (fig. 1H). Large records, especially in Pandanaceae and Triuridaceae. Our ob-
fleshy fruits are often highly vascularized in monocots, as in servations on Cyclanthus are inconclusive; no tapetum is vis-
some palms (e.g., Romanov et al. 2011), but this is not always ible at the tetrad stage, orbicules are absent, and the anther
the case; for example, the large fleshy fruits of some bamboos locule is filled with cytoplasmic material. Ultrastructural stud-
possess relatively few dorsal carpellary bundles (Rudall and ies are required to investigate this feature further.
Dransfield 1989). Similarly, Carludovicoideae possess rela- In all Cyclanthaceae, the ovary lacks septal nectaries, as in
tively highly vascularized tepals on female flowers, though the all other Pandanales except Velloziaceae, and is slightly sunken
perianth lobes of Cyclanthus are unvascularized. in the inflorescence axis. The ovary of Cyclanthaceae is uni-
Anther dehiscence is latrorse in Cyclanthaceae, compared locular with parietal placentation, as in the Sumatran genus
with relatively variable dehiscence among other Pandanales, Pentastemona (Fukuhara et al. 2003), which is sister to other
ranging from latrorse in Triuridaceae and introrse in Stemon- Stemonaceae (Mennes et al. 2013).
aceae to both latrorse and introrse in Velloziaceae (Rudall et
al. 2005; Rudall 2008; Sajo et al. 2010). Both anther wall Subfamilial Differences and Organ Interpretations
structure and tapetum type are unusually variable within Pan-
danales, though confusion has arisen from contrasting inter- Cyclanthus differs from other Cyclanthaceae in some re-
pretations. Most records are for the secretory tapetum type; spects. For example, the outer integument is poorly developed
824 INTERNATIONAL JOURNAL OF PLANT SCIENCES

Fig. 9 Cyclanthus bipartitus (Cyclanthoideae; LM). A, Longitudinal section through adjacent staminate and pistillate cycles (as in fig. 8C),
with dashed lines indicating approximate regions of transverse sections. B–E, Serial transverse sections as indicated in A. an p anther, dcb p
dorsal carpellary bundle, fil p filament, fusfil p fused filaments, mc p mucilage canal, ov p ovules, pl p perianth lobe, stb p staminode
bundle, stde p staminode, stig p stigma, sty p stylar canal, vcb p lateral carpellary bundle. Scale bars p 200 mm in A; 100 mm in B–E.

in Cyclanthus, so that it appears unitegmic in places, and the fused together at their bases. However, in Carludovicoideae
endothecium is relatively poorly developed. the stamens form a small stalk containing a single concentric
The most striking difference between Cyclanthus and Car- vascular bundle that branches to supply individual stamens.
ludovicoideae is that the floral organs are arranged in cycles This compound staminal structure can be compared with either
around the inflorescence axis in Cyclanthus rather than being the androphore that occurs in staminate floral units of some
grouped into discrete units. In Cyclanthus, all parts of each Triuridaceae (Maas-Van de Kamer 1995; Rudall 2003) or the
pistillate cycle are laterally fused; rows of perianth lobes and stamen fascicles that occur in some Vellozia species (Sajo et
staminodes border the gynoecium, which forms a single ovary al. 2010). Despite their close similarity with stamen fascicles,
locule sunk into the inflorescence axis. Because of this high based on their locations, the staminal clusters in Carludovi-
aggregation, it is difficult to identify individual flowers. How- coideae are probably correctly interpreted as male flowers (e.g.,
ever, Wilder (1981) noted that some discrete units (both sta- Harling 1958; Eriksson 1994a). Groups of male flowers sur-
minate and pistillate) are discernible at very early develop- rounding a central female flower form a partial inflorescence.
mental stages, though they are often arranged in long rows of It appears likely that male flowers of all Cyclanthaceae lack
organs rather than flowerlike structures. He observed that in perianth lobes. The staminate cycles of Cyclanthus lack peri-
pistillate units, the perianth organs are delimited first by a anth lobes entirely. Some authors have interpreted as perianth
furrow, followed by the staminode primordia, which form as lobes the sterile lobes that are present on the androphore of
rows of protuberances parallel to the perianth, and, subse- all Carludovicoideae except Evodianthus (e.g., Harling 1958;
quently, the carpel primordia develop in a row. Staminode Harling et al. 1998). However, our observations make this
primordia are initiated after the stamen primordia have de- interpretation unlikely; we interpret these (clearly secretory)
veloped on adjacent staminate units. structures as staminodes. If they were perianth lobes, it appears
In both Cyclanthus and Carludovicoideae, the stamens are more likely that they would be inserted at the base of the
 Fig. 10 Cyclanthus bipartitus (Cyclanthoideae; LM). A, Transverse section (TS) axis at level of ovary, showing mucilage canals. B, TS sterile
tip of staminode. C, TS anther. D–F, Sections of ovules. G, TS perianth lobe, showing row of large crystal idioblasts. anl p anther locule, ci p
crystal idioblast, dcb p dorsal carpellary bundle, es p embryo sac, ii p inner integument, mc p mucilage canal, oi p outer integument, ov p
ovule, pl p perianth lobe, vb p vascular bundle, vcb p lateral carpellary bundle. Scale bars p 100 mm in A, C, G; 50 mm in B, D–F.
826 INTERNATIONAL JOURNAL OF PLANT SCIENCES

androphore. Our observations show that they are inserted at dict these possible inferences, which require further testing. In
the same level as the filaments and possess similar vasculature. Liliales, loss of septal nectaries is correlated with a high degree
of congenital carpel fusion (Remizowa et al. 2010), but there
Conclusions is no such clear correlation in Pandanales. Within Cyclantha-
ceae, the anomalous (pseudanthial) reproductive structures of
Based on a morphological cladistic analysis, Rudall and Cyclanthus show some features in common with Carludovi-
Bateman (2006) postulated an evolutionary loss of develop- coideae, especially the presence of large axial mucilage canals,
mental homeostasis within Pandanales at the phylogenetic but also indicate a loss of developmental homeostasis. Our
node above Velloziaceae, leading to some anomalous floral observations considerably enhance existing knowledge of flo-
morphologies in the other four families, including complete ral anatomy of Cyclanthaceae and will provide a comparative
loss of septal nectaries, loss of trimery, and (in some clades) basis for future studies of Pandanales.
loss of perianth and development of unisexuality/dioecy. Some
authors have noted that loss of a perianth can result in insta-
bility in organ number and position of stamens and carpels Acknowledgments
(e.g., Endress 1990). Overlap in inflorescence and flower ge-
netic programs could have resulted from a partial loss of in- We thank Dr. E. W. (Ted) Taylor for the photographs of
tegrity in the regulation of flower development, resulting in Carludovica palmata in figure 1. M. G. Sajo, J. A. Lombardi,
(perhaps iterative) formation of pseudanthial structures. The and R. C. Forzza thank CNPq for the fellowship grant. J. A.
well-sampled molecular analysis of Mennes et al. (2013), Lombardi also thanks FAPESP and MCT/CNPq/MEC/CAPES/
though differing in some details of topology, does not contra- PROTAX for financial support.

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