Anatomia Frutos Mono PDF

RESEARCH ARTICLE
A M E R I C A N J O U R N A L O F B O TA N Y
Anatomy of fleshy fruits in the monocots1

Marcela Thadeo2,3,5, Katherine E. Hampilos2,4, and Dennis W. Stevenson2
PREMISE OF THE STUDY: An anatomical and developmental study of distantly related fleshy fruits in the monocots was undertaken to better understand the
evolution of baccate fruits in the monocot clade as a whole. We studied 14 species with fleshy fruits spanning the Alismatales, Arecales, Asparagales, Com-
melinales, Dioscoreales, Liliales, and Poales to determine various mechanisms through which baccate fruits attain fleshiness at maturity.
METHODS: Flowers and fruits of various stages were collected, sectioned, stained, and examined using light microscopy and scanning electron
microscopy.
KEY RESULTS: Three basic pathways for attaining fleshiness were identified within the species examined (true berries, with a uniform pericarp; typical
drupes, with an endocarp differentiated by the presence of stony pyrenes; and specialized drupes, involving mesocarp and endocarp differentiated by
stone pyrenes). Furthermore, developmental characters differentiating basic fruit types were identified.
CONCLUSIONS: Fleshy fruits in the monocots do not develop through a single shared pathway, indicating that fleshiness has evolved multiple times within
the clade.
KEY WORDS berries; capsules; drupes; fruit anatomy; fruit development; monocots
The monocot clade, containing over 65 000 species across 12 orders, evolution. Indeed, phylogenetic optimization of capsule-to-berry
is one of the most morphologically varied and ecologically success- transformations in the monocots suggests that these transitions
ful of the early-divergent clades of angiosperms (Givnish et al., have occurred more than 20 times (Givnish et al., 2005).
2006). As a result of the group’s diversity, a cohesive understanding The independent evolution of berries multiple times in the
of monocot phylogeny and patterns of morphological divergence monocot clade leaves open the possibility that separate lineages with
has not yet been reached. In particular, fruit development is rela- baccate fruits may have different developmental pathways. How-
tively poorly understood in the monocot clade. The establishment ever, detailed analyses of fruit morphology and development for
of an adequate fruit typology for the monocots will be useful not those monocot families are often lacking. In 2006, the initial results
only in understanding the evolution of the trait itself, but will also be of large-scale comparative study were published by Rasmussen
applicable to studies in ecology, conservation biology, and genetics. et al. (2006) with the intention of contributing to an ongoing project
Capsules are generally accepted to be the ancestral fruit type for to “reveal homologies and establish adequate fruit typology”. Focus-
angiosperms, from which berries and drupes evolved (Dahlgren ing on the Asparagales clade, Rasmussen et al. conducted develop-
et al., 1985). Fossil and molecular evidence supports this hypothesis mental studies on eight species from the Hyacinthaceae, Ruscaceae,
(Knapp, 2002). As a result, though the chosen outgroups in most and Amaryllidaceae, specifically choosing species with capsular
phylogenetic analyses of the monocot clade show a variety of fruit and baccate fruit types for their comparative analysis. In their con-
types, it is assumed that the ancestral character state for the clade is clusion, they noted that while the berries of the species they ana-
capsular and that the presence of baccate fruits (fleshy, indehiscent lyzed had relatively little structural variation, other berries from the
fruits) in various orders and families is the result of convergent liliflorous monocots (defined by Rasmussen et al. as the orders As-
paragales, Liliales, Dioscorales, and Pandanales) had different ways
1
Manuscript received 6 May 2015; revision accepted 21 September 2015. of attaining succulence and various mechanisms for partial opening.
2
The New York Botanical Garden, 2900 Southern Blvd., Bronx, New York 10458-5126 USA; We therefore aimed to expand the scope of the study initiated by
3
Universidade Estadual de Maringá, Av. Colombo, 5790, Maringá, Paraná 87020-900 Rasmussen et al. by examining the anatomy and development of
Brazil; and
4
Department of Biology, Oberlin College, 173 W Lorain St, Oberlin, Ohio 44074 USA
flowers and fruits from select species with baccate fruit not only in the
5
Author for correspondence (e-mail: [email protected]) Asparagales, but also in all monocot orders except one (Zingiberales)
doi:10.3732/ajb.1500204 in which fleshy fruits are known (Fig. 1). The study of fleshy
A M E R I C A N J O U R N A L O F B OTA N Y 102(11): 1757–1779, 2015; http://www.amjbot.org/ © 2015 Botanical Society of America • 1757
1758 • A M E R I C A N J O U R N A L O F B OTA N Y
FIGURE 1 Strict consensus tree for monocotyledons generated using a combined data set with morphological and molecular data.
N O V E M B E R 2015 , V O LU M E 102 • T H A D E O E T A L. — A N ATO M Y O F F L E S H Y F R U I T S I N T H E M O N O COT S • 1759
fruits in Zingiberales and arrilate seeds needs to be done in the ethanol. Some fruit and flower samples were dissected and photo-
future. graphed using a stereoscopic microscope. Additional fruit and
This study will focus on detailed observations of pericarp flower samples of various stages were prepared for embedding in
ontogeny in select species to understand and compare the various glycol methacrylate (Technovit H7100, ΕΞΑΡΤ−Apparatebau
potential methods of distantly related baccate fruits for attaining GmbH, Norderstedt, Germany) using the methodology proposed
succulence. The range of variation in the pericarp of baccate fruits for herbarium material (Meira and Martins, 2003) to insure pene-
has not been known (Rasmussen et al., 2006), and most informa- tration and sectioned using a rotary microtome. Also to insure pen-
tion on fruit anatomy consists of descriptive studies of external etration, the flowers and fruits were cut in half previously to
morphology that do not take into account the development of peri- embedding techniques. Serial sections (4–6 μm thick) were stained
carp tissues (Leite et al., 2009). Such descriptions of mature fruit with toluidine blue pH 4.0 (O’Brien and McCully, 1981) for struc-
morphology can be deceptive because of the range of pericarp tex- tural characterization. Qualitative histochemical reactions were
ture possible in both berries and capsules. For example, leathery employed on select sections to determine the origin and composi-
capsules and soft-walled irregularly dehiscing fruits can only be tion of various compounds using Coomassie brilliant blue (Fisher,
distinguished by the degree of lignification of layers in the pericarp 1968) for total proteins, ruthenium red for pectic substances
tissue (Roth, 1977). By examining the development of the pericarp (Johansen, 1940), phloroglucinol for lignin (Johansen, 1940), Su-
in fleshy fruits from various monocot orders, we aimed to provide dan IV for total lipid (Pearse, 1980), and periodic acid–Schiff re-
accurate information on the development of fleshy fruits in the agent (PAS) for total polysaccharides (Maia, 1979).
monocots to better understand the evolution of fruit types in the Samples were also observed with a scanning electron micro-
monocot clade on a larger scale. scope (SEM). For this process, samples were dried using hexameth-
It is also worth highlighting that some fruits, as in Acorus, are yldisilazane (HMDS) (Araujo et al., 2003; Jeger et al., 2009) and
anatomically berries but not functionally fleshy fruits. They have coated with a thin layer of gold (Bozzola and Russel, 1992).
dry coats and lack the sweet or oily composition that is usually as- Flowers at various developmental stages were examined to un-
sociated with vertebrate dispersal (Givnish et al., 2005). Other fea- derstand the origin of pericarp and seed layers. Flowers of Joinvillea
tures of Acorus morphology and its geographical pattern of genetic ascendens and Smilax hispida were not observed directly because of
variation strongly implicate water dispersal (Liao and Hsiao, 1998). the lack of available specimens. Anatomical observations were re-
corded, and character states were scored according to the charac-
ters used for the Monocot Tree of Life Project (2007). A character
MATERIALS AND METHODS state matrix was generated for use in this project (morphological
matrix of 230 characters with 309 apomorphic states for 276 mono-
Selected fruits and flowers were collected at different botanical gardens cot genera, compilation of our data and from the literature). The
in the United States and during field trips to Australia (Table 1). morphological characters, including details about character scor-
Samples for dissection were fixed in FAA 50 (formalin–acetic acid– ing and character analysis, will be the focus of a separate paper, but
50% alcohol, 1:1:18) under vacuum and transferred to 50% ethanol they were mostly derived from Stevenson and Loconte (1995). This
for storage (Johansen, 1940). Flowers and fruits representing vari- matrix has been combined with molecular data from the nuclear,
ous stages of development were selected and dehydrated to 100% mitochondrial, and chloroplast genomes (rbcL, matK, atp1, nad5)
TABLE 1. List of investigated species and specimens. Herbarium abbreviations: AD (State Herbarium of South Australia); NY (The New York Botanical Garden);
and FTG (Fairchild Tropical Botanic Garden). Other species are cultivated in ABG (Adelaide Batanical Garden); MBC (Montgomery Botanical Center); UCB
(University of California Botanical Garden); and NYBG NG (The New York Botanical Garden Nolen Greenhouse).
Species Family Order Specimen details
Triglochin procera R.Br. Juncaginaceae Alismatales AD JGC3082A
Potamogeton crispus L. Potamogetonceae Alismatales NY 01132302
NY 02342547
Ruppia maritima L. Ruppiaceae Alismatales FTG 99823
AD JGC3086
Zannichellia palustris L. Zannichelliaceae Alismatales NY 01132304
NY 02342541
Chamaedorea seifrizii Burret Arecaceae Arecales MBC 93-87A
Dianella caerulea Sims. Hemerocallidaceae Asparagales NYBG NG 81/2007*A
Geitonoplesium cymosum (R.Br.) R.Br. Hemerocallidaceae Asparagales UCB
Cordyline stricta (Sims) Endl. Laxmanniaceae Asparagales ABG G875801
Sowerbaea juncea Sm. Laxmanniaceae Asparagales ABG JGC3090
Palisota barteri Hook. Commelinaceae Commelinales NYBG NG 507/45*A
Tacca chantrieri André Taccaceae Dioscoreales NYBG NG 4100/95
Smilax hispida Raf. Smilacaceae Liliales NY 01132551
NY 01116545
Aechmea aquilega Griseb. Bromeliaceae Poales NYBG NG1411/2007*A
NYBG NG1411/2007*B
Flagellaria indica L. Flagellariaceae Poales NY 202024
Joinvillea ascendens Gaudich. Ex Brongn. & Gris Joinvilleaceae Poales NYBG NG 800-379
to generate the tree in Fig. 1. The phylogenetic parsimony analysis Regarding the monocotyledons genera scored in this study,
was conducted in the program TNT version 1.1 (Goloboff et al., fleshy fruits are found in Acoraceae, 23 terminals of the 33 Alisma-
2007). TNT was run using the ratchet (Nixon, 1999) with 200 inter- tales, 2 in Dioscoreales (9), 6 in Pandanales (11), 8 in Liliales (20),
actions of 5 up and 5 down, 1000 replicates, and 10 trees held per 15 in Asparagales (60), all terminals in Arecales (29), 2 in Commeli-
replicate. The branch collapsing rule was set to amb- (zero length nales (16), 3 in Zingiberales (14), and 4 in Poales (61). Fleshiness
branches are collapsed when their minimum lengths are zero). All represents 36% of the terminals scored (Fig. 1).
characters were weighted equally and treated as nonadditive, and Specific observations of gynoecium structure taken from prean-
gaps were treated as missing during the tree search. Trees were thesis and postanthesis carpels as well as mature fruits are included
viewed and characters were mapped using the program Winclada here to understand the origin and development of all cell layers in
(Nixon, 2002). Character states of relevance for this paper are de- the mature pericarp. Observations are organized on the level of or-
scribed in Appendix 1. der to highlight structural similarities between more closely related
Fruits of varying developmental stages were examined for all species and to understand anatomical differences on a broad scale.
taxa selected (Table 1). Characters delimiting the cell number,
shape, type, and content in various layers of the pericarp were cho- Alismatales—Triglochin procera (Juncaginaceae), Potamogeon cris-
sen and scored for each species examined (Appendix 2). pus (Potamogetonaceae), Ruppia maritima (Ruppiaceae), and Zan-
nichellia palustris (Zannichelliaceae).
Terminology applied to fruit typology and pericarp layers—For
the purpose of this study, we followed the indehiscent fleshy fruit Juncaginaceae—Triglochin procera. Flowers contain a superior,
typology proposed by Spjut (1994). This author based the fruit apocarpous gynoecium of six weakly connate carpels (Fig. 2A).
types on pericarp texture: berries have a uniform and undifferenti- Each carpel is unilocular (Fig. 2B) and contains a single bitegmic,
ated pericarp (characterized by a complete lack of sclerified cells anatropous, endostomic ovule. The fruits are schizocarpic, with
and an uniformly parenchymatous pericarp); and drupes have a each mericarp separating from the central columella (axis where
pericarp internally differentiated by one or more stony pyrenes. the carpels are attached) at maturity and forming a drupe, contain-
The term pyrene, according to Romanov et al. (2011), is used to ing a single seed (Fig. 2C).
indicate the stony zone of the pericarp, which may be of either en-
docarpic or mesocarpic origin. Preanthesis—The carpel tissue is made up of a uniseriate inner and
The technical boundaries used to delimit various layers of the outer epidermis and parenchymatous ground tissue in between.
pericarp can vary according to the observer’s interpretation. In the The intervening parenchymatous tissue ranges from 4 layers thick
broad sense, exocarp and endocarp are interpreted as containing near the central columella up to 16 layers thick to the lateralmost
epidermis and hypodermis (differentiated subepidermal tissue), portion of each carpel. The dorsal vascular bundles are directly ad-
while in stricter interpretations, exocarp and endocarp are delim- jacent to the endocarp, while there are 7–8 cell layers to the inside
ited by layers that originate from the epidermis layer itself (Roth, of the lateral vascular bundle (Fig. 2A). All intervening parenchyma
1977). A strict interpretation of pericarp layers is used in this study cells undergo prevalent anticlinal divisions. As the ovary matures,
to examine the development of the pericarp layers from a point of periclinal division also occurs in the tissue inside the vascular bun-
view of their origin. dles. The wings of the carpels, present midway between the central
Previous workers on fruit development have noted that within columella and the tip, contain numerous intercellular spaces in
the pericarp tissue, the location of the vascular bundles, which are cross section, presumably to keep the flower afloat in aquatic envi-
present at all stages of fruit development in all fruit types, can ronments (Fig. 2B). The testa and tegmen are both parenchymatous
constitute an additional useful boundary in comparative studies and are each 2 cell layers thick.
(Farmer, 1889; Pabón-Mora and Litt, 2011). In the present study,
the location of the vascular bundles within the pericarp was used Postanthesis—In the young fruit, the exocarp and endocarp are
to differentiate inner mesocarp (the cell layers between the endo- both uniseriate and parenchymatous. The number of cell layers in
carp and vascular bundles) from outer mesocarp (the cell layers the mesocarp has not increased—with no inner mesocarp layers
between the vascular bundles and the exocarp). The location of the and 4 outer mesocarp layers at the central columella, and up to 8
vascular bundles has been shown to form a zone where differenti- inner mesocarp layers and 8 outer mesocarp layers to the most lat-
ated tissues often meet (Pabón-Mora and Litt, 2011); hence, the eral portion of each carpel. Though the number of cell layers in the
use of this extra subdivision of the pericarp tissue allows us not mesocarp has not increased, mesocarp cells have expanded and in-
only to better understand fruit development within a single spe- tercellular spaces have increased in number and size. These spaces
cies, but also to more effectively compare pericarp textures across are concentrated in the outer mesocarp near the tip and the inner
species. mesocarp near the central columella.
In the mature fruit, the endocarp divides periclinally and is
sclerified, forming a biseriate pyrene (Fig. 2D). Each of the 6 meri-
RESULTS carps split from the central columella through the central vascular
bundles. The individual mericarps remain intact and do not de-
The parsimony tree shown in Fig. 1 is a strict consensus tree of the hisce. In the most lateral portion of the mericarp, a biseriate layer of
15 984 equally parsimonious trees found in the analysis with each sclerified cells extends from the endocarp to the vascular bundle
tree having 85 540 steps with a CI = 0.36 and a RI = 0.66. The phy- (Fig. 2D).
logeny calculated from the combined analysis places Acorus within
the Alismatales, and this clade is placed as sister group to all other Potamogetonaceae—Potamogeton crispus. Each flower contains a
monocots. superior, apocarpous gynoecium of 4 free carpels. Each carpel is
FIGURE 2 Triglochin procera (cross sections / A, B, and D, toluidine blue). (A) Gynoecium with six weakly connate carpels. (B) Unilocular carpel with a
single ovule. (C) Schematic drawing of the fruit. (D) Detail of the central portion of the mericarp, showing sclerified cells in the endocarp. Abbreviations:
dv, dorsal vascular bundle; en, endocarp; ie, inner epidermis; is, intercellular space; lv, lateral vascular bundle; o, ovule; oe, outer epidermis; ow, ovary
wall; s, seed.
unilocular and contains a single bitegmic, pendulous, endostomic parenchyma cells and vascular tissue and contains well-developed
ovule (Fig. 3A). The mature fruit is a drupe containing a single seed air spaces concentrated in the outer layers of the ovary apex. The
(Fig. 3B). single vascular bundle is nearly adjacent to the inner epidermis
(Fig. 3A).
Preanthesis—The carpel tissue contains a uniseriate outer epider- As the ovary matures, expansion as well as anticlinal division
mis of elongated tabular cells with a thick outer cuticle. The uniseri- can be seen in the cell layers outside the vascular bundles. The inner
ate inner epidermis is composed of relatively small tabular cells. epidermis can be seen undergoing extensive periclinal division, es-
The ground tissue between these layers is made up of isodiametric pecially toward the apex of the ovary.
FIGURE 3 Potamogeton crispus (A and C, cross sections; B and D, longitudinal sections / toluidine blue). (A) Detail of one unilocular carpel, with a single
ovule. (B) Total view of the fruit, highlighting the stony endocarp. (C, D) Details of the pericarp. Abbreviations: en, endocarp; ex, exocarp; ie, inner epi-
dermis; m, mesocarp; oe, outer epidermis; v, vasculature.
Postanthesis—The exocarp consists of a single layer of isodiamet- Postanthesis—The exocarp of young fruits consists of a single layer
ric cells with interlocking cell walls. These cells contain a dark- of cells with thick primary walls, a thin outer cuticle, and contain-
staining substance of unknown composition (Fig. 3C and 3D). The ing phenolic compounds and starch grains. As the fruit matures,
outer mesocarp contains relatively large parenchyma cells and air exocarp cells divide periclinally to form a biseriate layer. The outer
spaces, while the inner mesocarp contains only a few layers of mesocarp consists of ground tissue made of 2–4 layers of small iso-
small, thin-walled parenchyma cells that appear to be com- diametric parenchyma cells that are also filled with starch grains.
pressed at maturity. The endocarp is 2–7 layers of tabular scleri- The single medial vascular bundle lies directly adjacent to the endo-
fied cells (Fig. 3C and 3D). When stained with phloroglucinol, carp, and no inner mesocarp cells exist. Early in fruit development,
only the endocarp was positive for lignin. Both the tegmen and the endocarp is a biseriate layer of tabular parenchyma cells. As the
testa are parenchymatous and compressed beneath the endo- fruit matures, the endocarp cells divide periclinally, enlarge, and
carp at maturity. become thickened. The mature endocarp is 4–6 layers of rela-
tively large sclerenchyma cells with an unusual lignification pattern.
Ruppiaceae—Ruppia maritima. Each flower contains a superior, While these cells do not polarize under plane polarized light, SEM
apocarpous gynoecium of four free carpels. Each carpel is unilocu- images show that surface of the endocarp is uniform, and the un-
lar and contains a single bitegmic, pendulous, endostomic ovule. usual thickenings of the endocarp cells occur entirely within the
The mature R. maritima fruit is a drupe containing a single seed. endocarp cell walls. In young fruits, the testa is 4 layers thick, while
the tegmen is 2–3 layers thick and compressed. As the fruit ma-
Preanthesis—The carpel tissue contains a uniseriate outer epider- tures, both testa and tegmen become compressed beneath the scler-
mis of elongated tabular cells filled with phenolic compounds. The ified endocarp.
uniseriate inner epidermis is composed of relatively small tabular
cells. The ground tissue between is made up of isodiametric paren- Zannichelliaceae—Zannichellia palustris. Flowers contain a supe-
chyma cells and a single vascular bundle and contains air spaces rior, apocarpous gynoecium of 1–8 carpels. Each carpel contains 1
that become well-developed air lacunae toward the base of the bitegmic, anatropous, pendulous ovule. The mature fruit is a drupe
gynoecium. containing a single seed.
Preanthesis—The ovary wall consists of a uniseriate outer epider- the pericarp, and the fruit may grow through expansion of the
mis and a uniseriate inner epidermis, separated by ground tissue parenchymatous cells in the outer mesocarp and exocarp alone.
containing isodiametric parenchyma cells and a single vascular The embryo is curved.
bundle. The ground tissue varies in thickness from 2 layers thick
at the base and middle of the ovary up to 6 layers thick at the apex Arecales—Chamaedorea seifrizii (Arecaceae). Flowers have a tri-
of the ovary. The medial vascular bundle is located adjacent to the carpellate, syncarpous gynoecium. The one-seeded fruit develops
inner epidermis. The testa is 4 layers thick and parenchymatous, from 1 of the 3 uniovulate carpels; seeds do not develop in the other
and the tegmen is 3 layers thick and parenchymatous. 2 carpels which remain small in the fruit. The mature fruit is a spe-
cialized drupe (endocarp is not sclerified but their cells are filled
Postanthesis—The pericarp of the mature fruit contains a uniseri- with silica giving the equivalent hardness, part of the mesocarp is
ate, parenchymatous exocarp. The outer mesocarp is composed of sclerified, and both contribute to form the pyrene).
isodiametric parenchyma cells and ranges from 2 layers thick at the
base and middle of the fruit to 6 layers thick at the apex — i.e., there Preanthesis—The carpel tissue contains a uniseriate outer epider-
is no growth in number of cell layers in the mesocarp from prean- mis made up of cells with distinct radial elongation as well as some
thesis to postanthesis. The vascular bundle is located adjacent to thickening of all cell walls. The inner epidermis is also uniseriate.
the endocarp, indicating that there is no inner mesocarp growth in The ground tissue between these layers contains isodiametric pa-
this fruit. The endocarp contains 1–2 layers of sclerified cells, renchyma cells 14–28 layers thick, as well as vascular bundles
forming the stony pyrene. There is very little visible division in (Fig. 4A). The vascular bundles are not directly adjacent to the inner
FIGURE 4 Chamaedorea seifrizii (cross sections / A, B, D–F, toluidine blue). (A) Flower. (B) Ovary wall detail. (C) Detail of fruit. (D) Pericarp detail. (E) Exo-
carp and hypodermis cells. (F) Endocarp aspect. Abbreviations: en, endocarp; es, endosperm; ex, exocarp; hy, hypodermis; ie, inner epidermis; m, me-
socarp; oe, outer epidermis; ov, ovary; p, pericarp; ra, raphids; s, seed; ts, testa; v, vasculature.
epidermis, but in some places are separated by only 1–2 layers of indicating the possible presence of interlocular septal nectaries
cells. Cells outside the vascular bundles are larger than those in- (Fig. 5B).
side the bundles, while the cells inside the bundles can be seen to
be dividing periclinally. Cells containing calcium oxalate raphids Postanthesis—The exocarp consists of a single layer of cells with
are present only in the ground tissue outside of the vascular thin primary walls and a thin outer cuticle (Fig. 5B and 5C). For
bundles. both species, the outer mesocarp consists of 7–9 layers of isodia-
metric parenchyma cells, while the inner mesocarp consists of 5–6
Postanthesis—The exocarp of the young fruit is uniseriate and cells layers of isodiametric parenchyma cells (Fig. 5C). Idioblast cells
are radially elongated; however, there is no increase in size com- containing calcium oxalate raphids are present, through infrequent,
pared with the preanthesis ovary. The outer mesocarp is up to 24 in the mesocarp of G. cymosum, but are absent in the mesocarp of
layers thick, while the inner mesocarp is only 15 cell layers thick; D. caerulea. The endocarp of both fruits is a single layer of small,
neither layer of the mesocarp has intercellular spaces. In the outer cuboidal cells (Fig. 5B and 5C). In young fruits, the outer integu-
mesocarp, cells have divided anticlinally and periclinally to main- ment is 3–5 cell layers thick with a differentiated outer epidermis;
tain their size, except for cells containing calcium oxalate raphids, the inner integument is only 2 cell layers thick and compressed.
which have expanded and are significantly larger than other cells. As the fruits mature, exocarp cells becomes thickened on their
The endocarp is a thin, uniseriate layer containing dark-staining outer periclinal walls and can also divide anticlinally and enlarge
nuclei (Fig. 4B). (Fig. 5D). In both species, the cells in the outer mesocarp divide
As the fruit reaches maturity, the exocarp stains dark green periclinally to form a final layer 10 cell layers thick, while the in-
for phenolic compounds and forms a thick outer cuticle (Fig. 4D ner mesocarp remains 5–6 cell layers thick. In G. cymosum, cells
and 4E). Sudan tests show that the cuticle and the seed were pos- of the outer mesocarp do not expand notably, while in D. caeru-
itive for lipids. The mesocarp cells and the seed grow simultane- lea the outer mesocarp cells expand isodiametrically. There are
ously. The outer mesocarp forms a 2–3-layer hypodermis beneath no intercellular spaces in the mature fruits of both species. The
the exocarp that also stains green for phenolic compounds (Fig. endocarp divides periclinally and anticlinally to form a biseriate
4D and 4E). In the outer mesocarp, parenchymatous cells have endocarp, which eventually becomes thickened on both periclinal
been compressed, possibly by the growth of the seed, which con- walls. As the fruits mature, the outer integument develops into the
stitutes the majority of the mature fruit (Fig. 4C). Only cells phytomelaniferous testa (Fig. 5E), while the inner integument re-
containing calcium oxalate crystals remain uncompressed. The mains parenchymatous. In G. cymosum, the inner integument is
inner mesocarp layers have become highly compressed and lig- degraded and compressed into a thin layer of tegmen surrounding
nified (Fig. 4F), forming a thin, hard layer. The endocarp re- the endosperm tissue; in D. caerulea, the inner integument develops
mains uniseriate at maturity, but the cells are filled with silica into a parenchymatous tegmen 2–5 cell layers thick.
crystals (Fig. 4F). Phloroglucinol tests show that only the vascu-
lar bundles and inner mesocarp layer test positive for lignin. The Laxmanniaceae—Cordyline stricta. Fruits are baccate and indehis-
mature seed coat is hard and appears to have adhered to the en- cent at maturity (Fig. 6A).
docarp layer, possibly through pressure applied by the growth of
the seed. Preanthesis—The carpel tissue is made up of a uniseriate inner and
outer epidermis, both composed of cuboidal cells, and between
Asparagales—Cordyline stricta (Laxmanniaceae), Dianella caeru- them ground tissue, composed of relatively large isodiametric pa-
lea (Hemerocallidaceae), Geitonoplesium cymosum (Hemerocalli- renchyma and vascular tissue. In C. stricta, there are 4–5 layers of
daceae), and Sowerbaea juncea (Laxmanniaceae). All species were ground tissue on either side of the median vascular bundles. Septal
reported to contain fleshy, indehiscent fruits at maturity except for slits are present in the mesocarp near the center of the ovary, indi-
S. juncea, which is known to have a dry, dehiscent fruit in the form cating the possible presence of interlocular septal nectaries. Cells
of a capsule. This dry fruit was examined for comparison purposes containing calcium oxalate crystals are present throughout the
and to better understand the specific characters differentiating dry ground tissue outside the vascular bundles.
and fleshy fruits.
All species observed in the Asparagales contain a superior, syn- Postanthesis—The exocarp is uniseriate at maturity, and the cells
carpous, and trilocular gynoecium. Each of the three locules con- remain cuboidal and parenchymatous. The mesocarp has under-
tains two staggered rows of 4–6 ovules, attached to the axial gone extensive expansion as well as periclinal and anticlinal divi-
placenta. The ovules are bitegmic, anatropous, and endostomic. sion, and at maturity contains 8–10 cell layers in both the inner and
Mature fruits contain 1–3 seeds, with at most 1 seed per locule. outer mesocarp layers (Fig. 6B). Calcium oxalate raphides, as well
as large intercellular spaces, are also present in both the inner and
Hemerocallidaceae—Dianella caerulea; Geitonoplesium cymosum. outer mesocarp layers. As the fruit matures, the uniseriate endo-
Both fruits are fleshy berries at maturity. carp becomes lignified and develops into a thin layer of scleren-
chyma with U-shaped thickenings (Fig. 6C and 6D). Phloroglucinol
Preanthesis—The carpel tissue is made up of a uniseriate inner and tests were positive for lignin in the endocarp only. The mature testa
outer epidermis, both composed of tabular cells, and between them is phytomelaniferous and hard (Fig. 6A), while the parenchyma-
ground tissue, composed of relatively large isodiametric paren- tous tegmen is degraded and compressed beneath the endosperm
chyma and vascular tissue (Fig. 5A). In both species, the median tissue.
vascular bundles are not adjacent to the inner epidermis—there are
4–6 cell layers on either side of the vascular bundles. Septal slits are Laxmanniaceae—Sowerbaea juncea. Fruits are hard and dehiscent
present in the mesocarp near the center of the ovary in both species, at maturity.
FIGURE 5 Geitonoplesium cymosum (cross sections / A–D, toluidine blue). (A) Trilocular gynoecium. (B) Young fruit. (C) Young fruit pericarp detail.
(D) Mature fruit pericarp detail. (E) Detail of fruit. Abbreviations: ao, abortive ovule; en, endocarp; es, endosperm; ex, exocarp; l, locule; m, mesocarp;
o, ovule; ow, ovary wall; p, pericarp; ph, phytomelanin; s, seed; ss, septal nectaries; v, vasculature.
Preanthesis—The carpel tissue is made up of a uniseriate inner and in longitudinal rows in the outer ovary wall and in horizontal
outer epidermis, both composed of cuboidal cells, and between planes in the septa (Fig. 7A).
them ground tissue, composed of relatively large isodiametric pa-
renchyma and vascular tissue. The median vascular bundles are ad- Postanthesis—The exocarp is uniseriate at maturity, although the
jacent to the inner epidermis, and only 2–3 layers of ground tissue cells have elongated circumferentially to form rectangular paren-
are present outside the vascular bundles at any point. Cells contain- chyma cells (Fig. 7B and 7C). The mesocarp has not increased in
ing calcium oxalate crystals are numerous in the ground tissue out- cell number—the median vascular bundles are adjacent to the en-
side the vascular bundles. These idioblasts are present in all floral docarp, and the outer mesocarp is only 2–3 cell layers thick. In the
organs but are more abundant in the ovary, where they often occur mature fruit, the uniseriate endocarp is lignified with U-shaped
FIGURE 6 Cordyline stricta (cross sections / B–D, toluidine blue). (A) Detail of fruit. (B) Mature fruit wall, highlighting fleshy parenquimatous mesocarp.
(C) Endocarp detail. (D) Endocarp detail under polarized light. Abbreviations: en, endocarp; ex, exocarp; m, mesocarp; p, pericarp; ph, phytomelanin;
s, seed.
thickenings (Fig. 7C). The sclerified endocarp cells shrink as they isodiametric parenchyma cells and vascular bundles (Fig. 8A). There
approach the area of the median vascular bundle and are not pres- are 8 layers of ground tissue to the outside of the vasculature and
ent immediately adjacent the vascular bundle (Fig. 7B). Over time, 4 layers to the inside. Clear anticlinal division can be seen in all
the outer epidermis of the testa differentiates into phytomelan- layers of the ground tissue.
producing cells (Fig. 7C and 7D), which eventually become the
hard phytomelan seed coat. The parenchymatous endotesta and Postanthesis—The exocarp is uniseriate with clear anticlinal divi-
tegmen degrade. As the fruit matures, the fleshy mesocarp and exo- sion (Fig. 8C). The mesocarp has grown to 40–45 cell layers thick
carp dehydrate, while the endocarp layer retains its shape due to the with visible anticlinal and periclinal division in all layers of the me-
thickenings. socarp, though the division of inner and outer mesocarp is difficult
to determine due to the proliferation of vascular bundles (Fig. 8B).
Dioscoreales—Tacca chantrieri (Taccaceae). The gynoecium is in- The endocarp is uniseriate and parenchymatous (Fig. 8D). There
ferior, tricarpellate, and eusyncarpous. The ovary is unilocular and are no intercellular spaces between cells, and cells in all pericarp
contains numerous pendulous ovules in the single cavity (Fig. 8A). layers have expanded little. The seed has 4–5 layers of testa and 1–2
The mature fruit is a berry. layers of tegmen, all of them parenchymatous. The exotesta is a
single layer of tabular cells, the mesotesta has 2 distinct layers of
Preanthesis—The gynoecium is composed of a uniseriate inner radiate cells, and the endotesta can have 1 or 2 layers of also tabular
and outer epidermis and ground tissue in between, composed of cells (Fig. 8E).
FIGURE 7 Sowerbaea juncea (cross sections / A–C, toluidine blue). (A) Trilocular gynoecium. (B) Detail of part of fruit wall and seed. (C) Pericarp and
testa. (D) Schematic drawing of fruit. Abbreviations: en, endocarp; es, endosperm; ex, exocarp; m. mesocarp; o, ovule; ow, ovary wall; ph, phytomelanin;
ra, raphids; v, vasculature.
Commelinales—Palisota barteri (Commelinaceae). The gynoecium The integuments are parenchymatous; the outer integument is 4
is superior, syncarpous, and trilocular. The ovules are bitegmic and cell layers, and the inner integument is 2 cell layers.
endostomic (Fig. 9A). The mature fruit is a berry, containing 1–3
seeds (Fig. 9B). Postanthesis—In the young fruit, the exocarp is uniseriate, com-
posed of small, uniformly tabular cells with a thickened outer peri-
Preanthesis—The carpel tissue is composed of a uniseriate inner clinal cell wall. The outer mesocarp consists of 5–10 layers of large,
and outer epidermis and parenchymatous ground tissue in be- isodiametric parenchyma cells that are tightly packed, as well as
tween. The carpel contains distinct median vascular bundles that idioblast cells containing calcium oxalate raphides (Fig. 9C). These
are located adjacent to the inner epidermal layer, with 5 cell layers idioblast cells are larger than the surrounding parenchyma cells and
of parenchymatous ground tissue located outside the vascular bundle. often occur in rows that extend the length of the fruit in longitudinal
FIGURE 8 Tacca chantrieri (A–D, cross sections; E, longitudinal section / toluidine blue). (A) Unilocular and tricarpellate ovary, with numerous ovules. (B)
Detail of parenquimatous mesocarpic cells. (C) Exocarp in detail. (D) Endocarp in detail. (E) Seed aspect. Abbreviations: en, endocarp; es, endosperm;
ex, exocarp; ie, inner epidermis; m, mesocarp; oe, outer epidermis; tg, tegmen; ts, testa; v, vasculature.
sections. The median vascular bundles are immediately adjacent to mucilage to be a carbohydrate (Fig. 9E). The inner epidermis of the
the endocarp, and no inner mesocarp layers develop in this fruit. testa becomes sclerified to form a hard layer surrounding the endo-
The endocarp is composed of a single layer of small, thin-walled sperm (Fig. 9D).
cuboidal cells (Fig. 9C).
As the fruit matures, the exocarp cells remain uniseriate but be- Liliales—Smilax hispida (Smilacaceae). Mature fruits are berries
come thickened on all walls. In the layers immediately adjacent to and contain 1–3 seeds (Fig. 10A).
the exocarp, periclinal and anticlinal divisions (Fig. 9C) create 2–3
layers of thick-walled hypodermal tissue in the mature fruit (Figs. Preanthesis—Data not available.
9D, 9E and 9F). Cells in the outer mesocarp grow mainly through
expansion and contain no intercellular spaces, remaining 5–10 lay- Postanthesis—The exocarp consists of a single layer of uniformly
ers thick at maturity. The endocarp remains uniseriate and paren- small, lignified cuboidal cells with a thin outer cuticle. Beneath the
chymatous. The mature testa is parenchymatous and 5 layers thick. epidermis is a hypodermis 2–3 cell layers thick, composed of
Beginning in the early stages of fruit development, the outer epider- slightly larger cells ranging from cuboidal to isodiametric as they
mis of the testa secretes purple-staining mucilage into the locular differentiate toward the mesocarp. The majority of hypodermal
space, which then fills the locules with thick mucilage at matu- cells are filled with phenolic compounds. In young fruits, both an
rity (Fig. 9D). Periodic acid–Schiff reagent (PAS) tests show this inner mesocarp of 4–5 cell layers and an outer mesocarp of 9–10 cell
FIGURE 9 Palisota barteri (cross sections / A, safranin; C, D, F, toluidine blue; E, periodic acid–Schiff reagent). (A) Trilocular gynoecium. (B) Image of fruit.
(C) Young fruit pericarp detail. (D) Pericarp and seed from maturing fruit. (E) Pericarp and seed from maturing fruit highlighting positive reaction for
mucilage into the locular space. (F) Pericarp of mature fruit. Abbreviations: en, endocarp; es, endosperm; ex, exocarp; hy, hypodermis; m, mesocarp;
mu, mucilage; o, ovule; ow, ovary wall; p, pericarp; ra, raphids; s, seed; st, starch; tg, tegmen; ts, testa; v, vasculature.
layers can be identified. Both inner and outer mesocarp are com- Poales—Aechmea aquilega (Bromeliaceae), Flagellaria indica (Fla-
posed of isodiametric parenchyma cells with no intercellular spaces. gellariaceae), and Joinvillea ascendens (Joinvilleaceae).
Cells in the outer mesocarp also contain extensive phenolic com-
pounds. Cells in the inner mesocarp contain idioblast cells with Bromeliaceae—Aechmea aquilega. The gynoecium is syncarpous,
calcium oxalate crystals and sometimes mucilage. The endocarp is inferior, and trilocular. There are numerous anatropic, bitegmic
uniseriate, composed of small cuboidal cells that are not substan- ovules per locule. The mature fruit is fleshy and indehiscent at ma-
tially thickened. In young fruits, the testa is composed of 10–15 turity, containing numerous seeds.
layers of parenchymatous cells that can contain idioblast cells
with calcium oxalate raphides, all surrounding a sclerified tegmen Preanthesis—The gynoecium is composed of a uniseriate inner and
(Fig. 10B). outer epidermis with ground tissue in between, made up of isodia-
As the fruit matures, the cuticle thickens. The hypodermis layer metric parenchyma cells and vascular bundles. There are 10–12 lay-
grows to 5–6 cell layers adjacent to the exocarp that contain pheno- ers of parenchyma cells surrounding the vascular bundles and
lic compounds (Fig. 10C and 10D). The cells in the hypodermal 22–24 layers inside the vascular bundles, with clear periclinal divi-
layer retain their original size, while the remaining cells of the outer sion present in all layers of ground tissue. Idioblast cells containing
and inner mesocarp have expanded isodiametrically. The cells of calcium oxalate raphides are also present throughout the ground
the uniseriate endocarp are compressed at maturity (Fig. 10E). As tissue.
the seed develops, the cells of the thick parenchymatous testa ex-
pand and are eventually degraded, while the cells of the tegmen re- Postanthesis—In the mature fruit, the exocarp remains uniseriate,
main hard and protect the seeds. and all walls have thickened. Adjacent to the exocarp are 5–6 layers
FIGURE 10 Smilax hispida (cross sections / B–E, toluidine blue). (A) Image of fruit. (B) Young fruit. (C) Mature fruit. (D) Exocarp and hypodermis detail. (E)
Endocarp detail. Abbreviations: en, endocarp; es, endosperm; ex, exocarp); hy, hypodermis; m, mesocarp; p, pericarp; ra, raphids; s, seed; tg, tegmen;
ts, testa; v, vasculature.
of hypodermal tissue that have become sclerified. Though the divi- Postanthesis—The exocarp is uniseriate, and the cells are radially
sion of inner and outer mesocarp is difficult to determine due to the elongated, with a thickened outer cell wall. Adjacent to the exocarp
proliferation of vascular bundles in this fruit, an inner mesocarp is one layer of hypodermal tissue (Fig. 11B and 11C). The outer
layer clearly separates the innermost vascular bundles from the en- mesocarp is 20 layers thick and composed of isodiametric paren-
docarp. Idioblast cells containing calcium oxalate crystals are pres- chyma cells containing starch grains (Fig. 11E). The cells are largest
ent in both layers of the mesocarp, and no intercellular spaces are toward the exocarp and smallest toward the inner mesocarp; there
present in the pericarp. The endocarp is biseriate with inner and are no intercellular spaces. The inner mesocarp layer is sclerified
outer periclinal walls notably thickened in both layers. (Fig. 11B). The vascular bundles are mostly at the outer edge of the
sclerified layer. The cells in the inner mesocarp are sclereids, as can
Flagellariaceae—Flagellaria indica. The gynoecium is superior, tri- be seen in cross section (Fig. 11D and 11E). It is impossible to visu-
carpellate, and syncarpous. There is a single, pendulous, orthotro- ally differentiate the endocarp from the inner mesocarp in the ma-
pous ovule per locule. The mature fruit is a specialized drupe (not ture fruit to determine cell number, but the endocarp is clearly
just the endocarp is sclerified, but also part of the mesocarp, and sclerified (Fig. 11B). The seed coat remains parenchymatous and
both contribute to form the pyrene) with a single seed. compressed at maturity.
Preanthesis—The gynoecium is composed of a uniseriate outer epi- Joinvilleaceae—Joinvillea ascendens. The mature fruit is a special-
dermis and a biseriate inner epidermis with dark-staining nuclei. ized drupe (the endocarp is not sclerified, the pyrene is formed by
Between these layers is ground tissue, composed of parenchyma the sclerification of the mesocarp) containing 1–3 seeds (Fig. 12A).
and vascular bundles. There are 5–6 cell layers of isodiametric pa-
renchyma outside the vascular bundles, and 1–2 cell layers of elon- Preanthesis—Data not available.
gated and compressed parenchyma separating the vascular bundles
from the inner epidermis. The integuments are 2 cell layers each of Postanthesis—The exocarp is uniseriate, isodiametric in shape.
parenchyma cells (Fig. 11A). The outer mesocarp is around 8 cell layers thick, while the inner
FIGURE 11 Flagellaria indica (cross sections / toluidine blue). (A) Detail of tricarpellate gynoecium. (B) Pericarp of mature drupe. (C) Palisade-like exo-
carp. (D) Vascular bundle in the inner sclerified mesocarp. (E) Same as D but under polarized light. Abbreviations: en, endocarp; sc, sclereids; ex, exo-
carp; hy, hypodermis; ie, inner epidermis; m, mesocarp; oe, outer epidemis; st, starch; v, vasculature.
mesocarp is 5 layers thick (Fig. 12B and 12C). In early stages of DISCUSSION
development, the endocarp cells (and possibly inner mesocarp
cells) have secreted a substance of unknown composition into Even though the inclusion of Acorus within the Alismatales has
the locule (Fig. 12B). In more mature stages of development, this been reported before (Qiu et al., 2000; Davis et al., 2006), this is a
substance forms a uniform layer (Fig. 12C). controversial result of the present analysis (Fig. 1). This subject is
As the fruit matures, the cells in the exocarp became sclerified. further discussed by Petersen et al. (2015).
Cells in the outer mesocarp have expanded isodiametrically but do
not divide, remaining 8 layers thick; cells in the inner mesocarp Comparative anatomical analysis of pericarp development—
have not expanded but are visibly dividing both periclinally and Location and extent of sclerification—Identification of fruit type based
anticlinally, forming more cell layers (Fig. 12D). There are no on gross morphological characters such as “fleshiness” or “indehis-
spaces between cells in the inner or outer mesocarp. The cells sur- cence” ignores underling structural details of pericarp texture that
rounding the vascular bundles in both the inner and outer meso- can provide a more accurate picture of fruit development and evo-
carp are lightly sclerified, forming a hard layer in the center of the lution (Pabón-Mora and Litt, 2011). The location, origin, and ex-
mesocarp. The endocarp is a single, sometimes double, layer of tent of sclerified cell layers within the pericarp are important
small, compressed parenchyma cells. The secreted substance be- characters in the identification of fruit type because they often de-
tween the endocarp and the seed coat is most likely responsible for fine the mechanism by which the fruit dehisces or remains indehis-
compressing the endocarp layer. The testa is hard, and the tegmen cent at maturity (Von Guttenberg, 1971). Indehiscent fleshy fruit
is a single layer of parenchymatous cells (Fig. 12D and 12E). types are delimited by Spjut (1994) based on pericarp texture—the
FIGURE 12 Joinvillea ascendens (cross sections / B–D, toluidine blue). (A) Fruit image. (B) Young fruit pericarp. (C) Maturing fruit pericarp. (D) Mature
fruit. (E) Scanning electron microscopy of mature fruit. Abbreviations: en, endocarp; es, endosperm; ex, exocarp; p, pericarp; s, seed; sc, secretion;
v, vasculature.
drupe pericarp is internally differentiated by one or more stony py- Fleshy fruits containing a sclerified endocarp and uniformly
renes, while the pericarp of the true berry is uniform and undiffer- parenchymatous pericarp were found primarily in fruits of the
entiated. The fruits examined in this study, while all “fleshy” and Alismatales (Potamogeton crispus, Ruppia maritima, Triglochin
“indehiscent”, exhibit a wide range of sclerification patterns that procera, and Zannichellia palustris). All of these species were previ-
may be useful in identifying underlying homologies among mono- ously described as producing achene-type fruits (Dahlgren and
cot species. Clifford, 1982; Tomlinson, 1982). However, the micrographs pre-
The phenotype of the true berry, characterized by a complete sented in this study are not consistent with the commonly accepted
lack of sclerified cells and a uniformly parenchymatous pericarp, definition of the achene—a dry fruit with undifferentiated pericarp
was found in species spanning the Asparagales (Dianella caerulea, closely adhered to the seed (Spjut, 1994)—and instead show ligni-
Geitonoplesium cymosum), Commelinales (Palisota barteri), Di- fied endocarp cells that can be differentiated from the parenchy-
oscoreales (Tacca chantrieri), Liliales (Smilax hispida), and Poales matous mesocarp and exocarp in both species. It is therefore more
(Aechmea aquilega) (Fig. 1). While cell wall thickenings were pres- accurate to classify them as drupes (Fig. 1). This kind of misinter-
ent for some species in the endocarp (A. aquilega, D. caerulea, G. pretation happens very frequently in fruit classification, and one
cymosum) and for others in the exocarp (A. aquilega, P. barteri, S. reason is that authors use only external morphology, as already
hispida), no lignified cells were found in any pericarp layers. noted for Potamogeton (Takaso and Bouman, 1984). A structural
analysis as well the ontogeny of fruits is very important to resolve Location of vascular bundles within the pericarp—Lignification of
this issue (Souza, 1984; Oliveira and Beltrati, 1993). the endocarp is a crucial part of a capsule’s opening mechanism
The mature fruits of Cordyline stricta (Asparagales) were also (Von Guttenberg, 1971), and the study by Rasmussen et al. (2006)
found to contain a uniseriate, sclerified endocarp beneath a fully revealed that the structure of the median vascular bundle is also
parenchymatous mesocarp and exocarp. However, there are nota- relevant to the dehiscence process. However, observations from
ble differences regarding the extent of the lignified cell layers within this study indicate an additional character that may be required for
the pericarp of C. stricta that separates it from the other drupes dehiscence—the specific position of the median vascular bundle
observed in this study. The C. stricta endocarp is composed of rela- within the pericarp. In the dehiscing fruit examined here, the me-
tively small, cuboidal cells that, while lignified, are enclosed by a dian vascular bundle is located directly adjacent to the lignified en-
parenchymatous mesocarp and exocarp up to 21 cell layers thick. docarp at the point of dehiscence. Conversely, most of the fleshy,
In contrast, the sclerified endocarps of the drupes observed in the indehiscent fruits examined here contain an inner mesocarp sepa-
Alismatales are consistently at least two cell layers thick and are rating the median vascular bundle from the endocarp. In previous
surrounded by a parenchymatous mesocarp and exocarp only 4–10 observations of fleshy capsules (fleshy, dehiscent fruits that differ
layers thick. Therefore, the relative thickness of the hardened endo- from capsules only in the extent of their mesocarp) of Ornithoga-
carp in the drupes of the Alismatales is large compared with the C. lum nutans (Hyacinthaceae) and Galanthus nevus (Amaryllida-
stricta drupe, possibly indicating different functions of the scleri- ceae), the median vascular bundle was also located directly adjacent
fied endocarp and likely a different evolutionary pathway. to lignified endocarp, despite the mesocarp being of similar thick-
A third and final sclerification pattern was observed in fruits ness to the indehiscent fleshy fruits examined here (Rasmussen
from the Arecales (Chaemadorea seifrizii) and Poales (Joinvillea as- et al., 2006). We observed three exceptions to this observation in
cendens and Flagellaria indica). The pericarp of these fleshy fruits is the fruits of R. maritima, Z. palustrus, and P. barteri, which remain
differentiated by a stony pyrene that derives, at least in part, from fleshy and indehiscent at maturity, but do not contain an inner
the inner mesocarp layer. In the case of F. indica, the pyrene in- mesocarp layer separating the median vascular bundle from the
cludes both a sclerified endocarp and a sclerified inner mesocarp, endocarp. These fruits may therefore demonstrate intermediate
which together form a single hard layer next to the parenchyma- phenotypes indicating a recent dehiscent ancestor or a possible
tous pericarp. In contrast, the stony pyrenes of J. ascendens and C. reversion to the dehiscent state.
seifrizii are both composed of a sclerified layer of inner mesocarp
adjacent to the endocarp that does not contain lignified cells, but Seed coat characteristics and pericarp texture—A correlation was
instead contains cells hardened with specialized substances. In C. observed in the fleshy fruits examined here between seed coat and
seifrizii, a single silica body is deposited in each cell of the endocarp. pericarp texture. Fruits with a sclerified seed coat usually had an
Similar deposition of silica has been found in other chamaedoreoid entirely fleshy pericarp, while those with soft seed coats often
palms; however, these other genera (Mascarena) exhibit at least some had hard pericarp layers. The hard seed coat is known to act as
sclerification of the endocarp in addition to silica deposition and thus protection for the embryo against mechanical and biological dam-
contain a more definite endocarp than Chamaedorea spp. (Murray, age after dispersal (Boesewinkel and Bouman, 1984). The associa-
1973). We could not determine the nature of the secretion present tion observed here may indicate that in the fruits where the seed
between the endocarp and the seed coat in J. ascendens because it coat is reduced or nonexistent at maturity, the protective func-
did not react with any histochemical reagents used in this study. tion has been transferred through evolution to the stony pericarp
(Danivlova et al., 1995). We were able to differentiate three seed coat
Cell division and expansion—Postanthesis, the pericarp layers groups within the fleshy fruits examined here. The first contains
grow in all fleshy fruits examined here, but the location and extent fruits with a soft seed coat and a hard endocarp layer (Alismatales
of cellular division and expansion varies among species. The mech- and F. indica), but in some cases, as for Flagellaria, the mesocarp
anism by which fruits grow and expand is closely linked with their can also be sclerified. The second group contains the remaining
mature fruit type; fleshy fruit growth is generally attributed to cell species that form stony pyrenes (C. seifrizii and J. ascendens), P.
division and cell expansion, and capsular fruit growth is attributed barteri, A. aquilega, and S. hispida, where the seed coat is sclerified
exclusively to cell division (Roth, 1977). Many of the fleshy fruits and the endocarp is soft (but the fruit may contain hard inner/outer
observed in this study grow via cell proliferation and cell expansion mesocarp). The third group includes fruits with phytomelanin de-
in the mesocarp. However, drupes observed in the Alismatales in- posited in the seed coat and usually a parenchymatous pericarp
crease in size after fertilization primarily through cell expansion. (Asparagales).
All four species examined exhibit some degree of anticlinal division
in preanthesis ground tissue; however, mesocarp cells do not con- Evolution of fruit type in the monocots—Fleshy fruits observed in
tinue to divide after fertilization, and the postanthetic pericarp this study attain succulence via three distinct mechanisms. These
grows primarily through expansion of cells in the outer mesocarp. pathways were defined based on the following general features of
In T. procera, P. crispus, and R. maritima, the endocarp divides pericarp structure: (1) the presence or absence of sclerification in
periclinally and enlarges in the postantheric fruit, which also con- various layers of the pericarp; (2) the relative amount of cell divi-
tributes significantly to growth in size of the mature drupe. All sion and expansion in various layers of the pericarp; (3) the loca-
fruits observed in the Alismatales contain intercellular spaces in the tion of the vascular bundles within the mesocarp.
ground tissue preanthesis; these spaces increase in number and size
to form well-developed air spaces in the mature fruits. These spaces True berries—The first and most widespread pathway observed in
are likely present in both pre- and postantheric tissue to facilitate this study is common to fruits in the Asparagales (D. caerulea, G.
growth and development of the fruits and flowers in an aquatic en- cymosum), Dioscoreales (T. chantrieri), Liliales (S. hispida), and
vironment (Tomlinson, 1982; Les and Haynes, 1995). Poales (A. aquilega). These fruits maintain an entirely parenchymatous
pericarp throughout development—though exocarp and endocarp hypothesis requires additional research on the anatomy of baccate
cells may become thickened as the fruit develops, no lignified sec- fruits in the monocots because few fleshy fruits have been exam-
ondary cell walls are present in any layers. Development occurs ined at this point and deviations from this basic developmental
mainly through extensive division and expansion of cells in the me- pathway may exist.
socarp layers, while the exocarp and endocarp remain 1–2 layers
thick throughout development and at maturity. All fruits examined Evolution of typical drupes—The four fruits from the Alismatales
in this group contain thick inner and outer mesocarp layers, with observed here each contain a sclerified endocarp surrounded by a
the vascular bundles separated from the endocarp by the thickened layer of thin parenchyma. Although comprising fewer species than
inner mesocarp layer. These fruits develop according to the typical are present in many angiosperms families, the subclass Alismatidae
pathway observed for true berries. represents the putatively monophyletic group of aquatic angio-
sperms and includes approximately 500 species, recognized either 2
Typical drupes—A second common developmental pathway was or 3 orders, 12–15 families, and 56 genera (Les and Haynes, 1995).
found in all fleshy fruits examined in the Alismatales (P. crispus, R. The fruit types present in the order include achenes, berries, cap-
maritima, T. procera, and Z. palustris). Fruits in this group contain sules, follicles, and intermediate fruit types described as “fleshy cap-
a sclerified endocarp, which may grow to be up to 5 cell layers thick, sules” (Kubitzki et al., 1998). Among monocotyledons, follicular
while cells in the mesocarp and exocarp remain parenchymatous fruits are most abundant in the Alismatales and were generally con-
throughout development. Growth occurs mainly through a small sidered to be the ancestral fruit type for the order (Dahlgren and
amount of division and expansion of cells in the outer mesocarp. Clifford, 1982) before the berry-fruited Araceae (Kubitzki et al.,
Fruits in this pathway do not develop an inner mesocarp layer—as 1998) were considered as part of the order (APG, 1998). Interestingly,
a result, the vascular bundles are located directly adjacent to the a review of the literature indicates that all fruits in the Alismatales
sclerified endocarp. This type of development is characteristic of contain a sclerified endocarp except Araceae and Groenlandia sp.
typical drupes. (Potamogetonaceae), both of which contain entirely parenchyma-
tous pericarps at maturity (Kubitzki et al., 1998; Tomlinson, 1982).
Specialized drupes—The final developmental pathway identified is Our research supports the hypothesis that the follicle is the an-
common to fruits examined in the Poales (F. indica and J. ascen- cestral fruit type for the core Alismatales (Fig. 1) because the fleshy
dens) and Arecales (C. seifrizii). These fruits are unique in that they fruits examined here exhibit more character states in common with
contain thick layers of sclerified cells within the mesocarp. These dehiscent follicles than with indehiscent berries. In addition to the
drupes contain a stony pyrene that is composed of sclerified meso- sclerified endocarp and reduced outer parenchymatous layers,
carp as opposed to endocarp tissue. Drupes in this group also often none of the four drupes examined develop an inner mesocarp layer
contain specialized cells in the exocarp or endocarp layers. beneath the vascular bundles. These key characters are found in de-
hiscent fruits and indicate a possible follicle-fruited ancestor.
Evolution of true berries—This strikingly similar developmental The aquatic habitat of most Alismatales families offers unique
pathway found in fruits spanning the Asparagales, Commelinales, environmental constraints that must be taken into account when
Dioscoreales, Liliales, and Poales was unexpected due to the phylo- considering dispersal mechanisms. All known marine and hydro-
genetic distance between these orders (Fig. 1). One explanation for phylous monocots are concentrated in the Alismatales (Li and
this phenomenon is the presence of a common ancestral fruit type Zhou, 2009). Indeed, the four fruits observed here each develop in
(the dry, dehiscent capsule) from which berries in these orders are an aquatic environment, either wholly submerged (P. cripsus, R.
all hypothesized to have derived (Dahlgren and Clifford, 1982; maritima, and Z. palustrus) or with an emergent inflorescence (T.
Dahlgren et al., 1985). However, the lack of even minor differentia- procera) (Kubitzki et al., 1998). These fruits are drupes based in
tion in the texture of the pericarp in species so distantly related also anatomy but not functionally fleshy fruits. To understand the dis-
implies that a single, adaptive developmental pathway for baccate persal mechanism for each of these fruits, one must therefore con-
fruits has been conserved over long time spans. These findings are sider the effect of an aquatic environment on dehiscence. For
consistent with those of Rasmussen et al. (2006) for two additional example, a dehiscent follicle, when grown in water or dispersed
species of baccaceous fruits from the Asparagales, Mainthemum into water, would not be able to achieve the necessary torsions be-
racemosum (Ruscaceae) and Haemanthus albiflos (Amaryllidaceae) tween the hard endocarp and soft outer layers required for dehis-
and those of Fagundes and Mariath (2010) for two species of bac- cence (Roth, 1977) due to the inability of the soft outer layers to
caceous fruits from the Poales, Aechmea calyculata (Bromeliaceae) dehydrate in water. Instead, the follicle would likely remain inde-
and Billbergia nutans (Bromeliaceae). hiscent, and the outer soft layers would eventually decay in the
Equally surprising were the few characters identified as differen- aquatic surroundings. This scenario, in fact, has been reported
tiating the development of baccate and capsular fruits. The capsular for each of the four fruits examined here (Kubitzki et al., 1998;
fruit examined here, of S. juncea, differs in its developmental path- Tomlinson, 1982; Singh, 1965). For example, the dispersal mecha-
way from baccate fruits through the relative thickness of the meso- nism of P. crispus was described by Kubitzki et al. (1998, p. 412) as
carp layers, lignification of the endocarp, and relative location of follows: “The fruits, after being released, float on the water for a
the median vascular bundles. These findings are in accordance with period of time… eventually the fruit sinks and the pericarp decays.”
those of Rasmussen et al., (2006) for other dry capsules in the It is possible therefore that the indehiscent drupaceous fruits exam-
Asparagales. The limited morphological differences between dry ined here are derived from follicles that lost dehiscence as an adap-
capsules and true berries, in conjunction with the conserved devel- tation to aquatic life. Rather than sharing a common ancestor, the
opmental pathways that exist for both fruit types in these orders, morphological character likely arose several times within the family
may imply that transitions between the two fruit types occur in distantly related groups, as is common in the Alismatales (Li and
more frequently than anticipated in these orders. However, this Zhou, 2009).
Evolution of specialized drupes—The third and final develop- Davis, J. I., G. Petersen, O. Seberg, D. W. Stevenson, C. R. Hardy, M. P.
mental pathway observed, characterized by sclerified layers in the Simmons, F. A. Michelangeli, et al. 2006. Are mitochondrial genes useful
mesocarp, was unique to drupes in the Poales and Arecales. Previ- for the analysis os monocot relationships? Taxon 55: 857–870.
Fagundes, N. F., and J. A. Mariath. 2010. Morphoanatomy and ontogeny of
ous to this research, drupes with a stony pyrene in the mesocarp
fruit in Bromeliaceae species. Acta Botanica Brasílica 24: 765–779.
had not been observed in any monocot families except for the
Farmer, J. B. 1889. Contributions to the morphology and physiology of pulpy
Arecales (Murray, 1973; Romanov et al., 2011; Reis et al., 2012; fruits. Annals of Botany 3: 833–847.
Mazzottini-dos-Santos et al., 2014). It is therefore possible that this Fisher, D. B. 1968. Protein staining of ribboned Epon sections for light micros-
is a pathway unique to the Areciflorae; however, additional ana- copy. Histochemistry and Cell Biology 16: 92–96.
tomical studies of fruit types in the monocots are needed to be sure. Furness, C. A., and P. J. Rudall. 1999a. Inaperturate pollen in monocotyledons.
Dahlgren and Clifford (1982) previously hypothesized that the International Journal of Plant Sciences 160: 395–414.
drupes of the Areciflorae may not be derived from capsules. This Furness, C. A., and P. J. Rudall. 1999b. Microsporogenesis in monocotyledons.
hypothesis makes sense in the context of our observations for the Annals of Botany 84: 475–499.
Arecales and Poales when considering the character state changes Furness, C. A., and P. J. Rudall. 2001. Pollen and anther characters in monocot
systematics. Grana 40: 17–25.
that are required for the transition between dehiscent and indehis-
Furness, C. A., and P. J. Rudall. 2006. The operculum in pollen of monocoty-
cent fruit types. For a capsule to evolve into this type of drupe
ledons. Aliso 22: 191–196.
would require three character state changes (loss of endocarp Givnish, T. J., J. C. Pires, S. W. Graham, M. A. McPherson, L. M. Prince, T. B.
sclerification, proliferation of the inner mesocarp layer, and sclerifi- Patterson, H. S. Rai, et al. 2005. Repeated evolution of net venation and
cation of mesocarp layer), while the changes needed for a true berry fleshy fruits among monocots in shaded habitats confirms a priori predic-
to evolve into this type of drupe would require only the sclerifica- tions: Evidence from an ndhF phylogeny. Proceedings of the Royal Society of
tion of mesocarp layer. It is therefore possible that the ancestral fruit London, B, Biological Sciences 272: 1481–1490.
type for these specialized drupes is a true berry. Givnish, T. J., J. C. Pires, S. W. Graham, M. A. McPherson, L. M. Prince, T. B.
Patterson, H. S. Rai, et al. 2006. Phylogenetic relationships of monocots
based on the highly informative plastid gene ndhF: Evidence for widespread
ACKNOWLEDGEMENTS concerted convergence. Aliso 22: 28–51.
Goloboff, P. A., J. S. Farris, and K. C. Nixon. 2007. Tree analysis using
The authors thank N. Pabón-Mora for helpful discussions, J. Conran new technology, version 1.1. Available at http://www.zmuc.dk/public/
for field assistance in Australia, and the anonymous reviewers phylogeny.
for their helpful and constructive comments and suggestions on Guo, Y., R. Sperry, C. D. K. Cook, and P. A. Cox. 1990. The pollination ecology
our manuscript. Research and study funds were kindly provided by of Zannichellia palustris L. (Zannichelliaceae). Aquatic Botany 38: 341–356.
The New York Botanical Garden, Oberlin College Department of Igersheim, A. I., M. Buzgo, and P. K. Endress. 2001. Gynoecium diversity and
Biology, and the NSF grant From Acorus to Zingiber — Assembling systematics in basal monocots. Botanical Journal of the Linnean Society 136:
the Phylogeny of the Monocots (DEB-082762). M.T. and K.E.H. 1–65.
Jeger, R., Y. Lichtenfeld, H. Peretz, B. Shany, R. Vago, and D. Baranes. 2009.
contributed equally to this work.
Visualization of the ultrastructural interface of cells with the outer and
inner-surface of coral skeletons. Journal of Electron Microscopy 58: 47–53.
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APPENDIX 1 Character states used in morphological and anatomical analysis of flowers and fruits of monocots at various developmental stages in order to
understand the origin of pericarp and seed layers. Characters were mostly derived from Dahlgren and Clifford (1982), Stevenson and Loconte
(1995), and this work. When another study was used, the reference is indicated.
Floral morphology
1. Floral expression: monoclinous (0), diclinous (1). Tomlinson (1982) / Remizowa et al (2010).
2. Flowers rubbish: solitary (0), inflorescence (1). Tomlinson (1982).
3. Tepals spotted: absent (0), present (1)
4. Flower fusion: hypogynous (0), perigynous (1), epigynous (2)
5. Deliquescent flowers: absent (0), present (1)
6. First tepal position: adaxial (0), abaxial (1). Buzgo and Endress (2000) / Buzgo (2001).
Perianth
7. Outer perianth: absent (0), present (1)
8. Outer perianth form: sepaloid (0), petaloid (1)
9. Outer perianth connation: absent (0), present (1)
10. Monosymetric outer perianth: absent (0), present (1)
11. Inner perianth: absent (0) present (1)
12. Inner perianth form: sepaloid (0), petaloid (1)
13. Inner perianth connation: absent (0), present (1)
14. Monosymetric inner perianth: absent (0), present (1)
15. Perianth curvature: absent (0), present (1)
16. Perigonal nectaries: absent (0), present (1). Smets et al (2000).
Androecium
17. Outer stamens: zero (0), one (1), two (2), three (3), many (4). Kubitzki et al (1998) / Remizowa et al (2010). Polymorphisms: Triglochin (2,3).
18. Monosymetric outer staminal whorl: absent (0), present (1). Kubitzki et al (1998).
19. Outer androecial whorl: epitepalous (0), hypogynous (1). Kubitzki et al (1998).
20. Adnate outer stamens: absent (0), present (1). Kubitzki et al (1998).
21. Outer staminodia: absent (0), present (1). Kubitzki et al (1998).
22. Outer connate stamen filaments: absent (0), present (1). Kubitzki et al (1998).
23. Inner stamens: zero (0), one (1), two (2), three (3), many (4). Kubitzki et al (1998) / Remizowa et al (2010). Polymorphisms: Triglochin (2,3).
24. Monosymetric inner staminal whorl: absent (0), present (1). Kubitzki et al (1998).
25. Inner androecial whorl: epitepalous (0), hypogynous (1). Kubitzki et al (1998).
26. Adnate inner stamens: absent (0), present (1). Kubitzki et al (1998).
27. Inner staminodia: absent (0), present (1). Kubitzki et al (1998).
28. Inner connate stamen filaments: absent (0), present (1). Kubitzki et al (1998).
29. Connective protusion: absent (0), present (1)
30. Anther attachment: basifixed (0), dorsifixed (1), ventrifixed (2), peltate (3)
31. Anther dehiscence orientation: introrse (0), extrorse (1), latrorse (2)
32. Anther dehiscence type: longitudinal (0), poricidal (1), valvar (2). Polymorphisms: Dianella (0,1).
33. Thecae per anther: two (0), one (1)
34. Microsporangia per theca: two (0), one (1)
35. Anther wall formation: dicotyledonous type (0), monocotyledonous type (1), basic type (2), reduced type (3)
36. Endothecial wall thickenings: spiral (0), girdle like (1), stellate (2), absent (3)
37. Exothecium: absent (0), present (1). Johri (1992).
38. Tapetum type: glandular secretory (0), plasmodial amoeboid (1). Furness and Rudall (1999b) / Furness and Rudall (2001).
39. Microsporogenesis: successive (0), simultaneous (1), modified simultaneous (2). Furness and Rudall (1999b) / Furness and Rudall (2001).
40. Pollen units prior to dispersal: monads (0), tetrads (incl.pseudomonads) (1), dyads (2), moniliform chains (3). Cox (1988) / Johri (1992). Polymorphisms:
Aechmea (0,1).
41. Extra pollen material (at shedding): absent (0), mucilaginous chains (1), mucilaginous tubes (2), massulae and pollinia (3). Cox (1988) / Johri (1992).
Polymorphisms: Ruppia (1,2).
42. Aperture number: one (0), two (1), three (2), four to many (3), none (4), convergent (5). Guo et al (1990) / Furness and Rudall (1999a). Polymorphisms:
Aechmea (0,3), Chamaedorea (0,1,2), Palisota (0,2), Smilax (0,4).
43. Porate pollen operculum: absent (0), present (1), coarse granules (2). Furness and Rudall (1999a) / Furness and Rudall (2006).
44. Exine: present (0), vestigial (1). Philbrick and Les (1996).
45. Exine protuberances: absent (0), spinulose-gemmate (1). Zavada (1983) / Guo et al (1990).
46. Scrobiculi: absent (0), present (1)
47. Pollen aperture margin: nonannulate (0), annulate (1)
48. Pollen wall ultrastructure: columellate (0), granular (1), aveolar (2), atectate (3)
49. Pollen aperture type: sulcate (0), synsulcate (1), porate (2), trichotomosulcate (3), ulcerate (4), clypeate (5), spiraperturate (6), cryptoaperturate (7),
omniaperturate (8), zonosulcate (9). Guo et al (1990) /Furness and Rudall (1999a). Polymorphisms: Smilax (0,5).
50. Tectum ornamentation: tectate (0), semitectate (1), intectate (2), atectate (3). Zavada (1983).
51. Endexine: absent (0), present (1). Zavada (1983) / Guo et al (1990).
52. Pollen nuclei: two (0), three (1). Guo et al (1990).
Gynoecium
53. Gynoecium position: superior (0), inferior (1). Rudall (2002) / Remizowa et al (2010).
54. Style fusion (in syncarpous taxa): absent (0), distinct (1). Remizowa et al (2010).
55. Stigmatic surface: dry (0), wet (1). Guo et al (1990) / Philbrick (1991) / Igersheim et al (2001) / Nunes et al (2012).
56. Stigma: symmetric (0), asymmetric (1)
57. Stigma branching: simple (0), branched (1). Tomlinson (1982) / Igersheim et al (2001).
58. Styles: solid (0), hollow (1)
59. Pseudomonomery: absent (0), present (1)
60. Carpel connation: apocarpous (0), syncarpous (1). Mering and Kadereit (2010).
61. Intraovarian mucilage: absent (0), present (1)
62. Ovules per locule: one (0), many (1)
63. Single ovule: basal (0), apical (1), median (2). Tomlinson (1982).
64. Placentation: laminar (0), marginal (1)
65. Syncarpous placenta: axile (0), parietal (1), free central (2). Tomlinson (1982) / Kubitzki et al (1998). Polymorphisms: Tacca (0,1).
66. Septal nectaries: absent (0), internal (1), external (2). Smets et al (2000) / Rudall (2002).
67. Nucellus: crassinucellate (0), tenuinucellate (1), pseudo-crassinucellar (2). Rudall (1997). Polymorphisms: Dianella (0,1).
68. Micropyle: endostomic (0), exostomic (1). Kubitzki et al (1998).
69. Zigzag micropyle: absent (0), present (1)
70. Embryostega: absent (0), outer integument (1), inner integument (2), both (3)
71. Embryosac starch: small grains (0), large grains (1)
72. Integument tannin: absent (0), present (1)
73. Ovule type: anatropous (0), campylotropous amphitropous (1), orthotropous (2). Mering and Kadereit (2010). Polymorphisms: Potamogeton (1,2).
74. Ovules: normal nucellus (0), massive nucellus (1)
75. Nucellus epidermal cells: elongated (0), square (1)
76. Nucellus shape: ovate (0), elliptical (1)
77. Nucellar cap: absent (0), present (1)
Fruit
78. Fruit type: follicle (0), achene (1), drupe (2), berry (3), utricle (4), capsule (5), schizocarp (6), nutlet (7), samara (8). Tomlinson (1982) / Romanov et al.
(2011).
79. Syncarpous fruit: dehiscent (0), indehiscent (1)
80. Fleshy exocarp: absent (0), present (1)
81. Fleshy mesocarp: absent (0), present (1)
82. Fleshy endocarp: absent (0), present (1)
83. Exocarp epidermis: absent (0), uniseriate (1), biseriate (2)
84. Exocarp hypodermis: absent (0), uniseriate (1), biseriate (2), three-five (3)
85. Exocarp cell shape: isodiametric (0), cuboidal (1), rectangular (2), compressed (3), elongated (4), irregular (5)
86. Exocarp cell type: parenchymatous (0), thick-walled (1), sclerified (2). Polymorphisms: Aechmea (1,2).
87. Exocarp phenolic idioblast: absent (0), small (1), large (2)
88. Sclerified mesocarp: absent (0), present (1)
89. Mesocarp raphide idioblast: absent (0), present (1)
90. Mesocarp phenolic idioblast: absent (0), present (1)
91. Endocarp epidermis: absent (0), uniseriate (1), biseriate (2)
92. Endocarp cell shape: isodiametric (0), cuboidal (1), rectangular (2), compressed (3), elongated (4), irregular (5)
93. Endocarp cell type: parenchymatous (0), thick-walled (1), sclerified (2)
94. Endocarp hypodermis: absent (0), uniseriate (1), biseriate (2)
95. Endocarp phenolic idioblast: absent (0), present (1)
Seed
96. Embryo sac development: monosporic (0), bisporic (1), tetrasporic (2)
97. Embryogeny: onagrad (0), caryophyllad (1), solanad (2), asterad (3), chenopodiad (4). Polymorphisms: Chamaedorea (0,3).
98. Endosperm in mature seed: absent (0), present (1)
99. Endosperm formation: cellular (0), nuclear (1), helobial (2). Swamy and Parameswaran (1963) / Mering and Kadereit (2010). Polymorphisms: Ruppia (1,2).
100. Helobial endosperm: normal (0), haemodore type (1)
101. Starchy endosperm: none or little (0), abundant (1)
102. Chalazosperm: absent (0), present (1). Rudall (1997).
103. Dermal perisperm: absent (0), present (1)
104. Subdermal perisperm: absent (0), present (1)
105. Phytomelan seeds: absent (0), present (1)
106. Microspermy: absent (0), present (1)
107. Aril: absent (0), present (1)
108. Testa cell type: compressed (0), parenchymatous (1), sclerified (2)
109. Tegmen cell type: compressed (0), parenchymatous (1), sclerified (2)
APPENDIX 2 Scores for the 109 ch aracter states used in morphological and anatomical analysis of flowers and fruits of monocots at various developmental stages in order to understand the origin
of pericarp and seed layers by taxon. Key: ?, unknown; —, inapplicable; *, polymorphic (see Appendix 1 for states).
Taxon 1–5 6–10 11–15 16–20 21–25 26–30 31–35 36–40
Aechmea 0 1 0 2 0 ? 1 0 0 0 1 1 0 0 0 0 3 0 0 0 0 0 3 0 1 0 0 0 0 0 0 0 0 0 1 0 ? 0 0 *
Chamaedorea 1 1 0 0 0 ? 1 0 0 0 1 0 0 0 0 0 3 0 1 0 0 0 3 0 1 0 0 0 0 0 2 0 0 0 1 0 ? 0 * 0
Cordyline 0 1 0 0 0 ? 1 1 0 0 1 1 0 0 0 0 3 0 1 1 0 0 3 0 1 1 0 0 0 1 0 0 0 0 1 1 0 ? ? ?
Dianella 0 1 0 0 0 ? 1 0 0 0 1 1 0 0 1 0 3 0 1 0 0 0 3 0 1 0 0 0 0 0 0 * 0 0 1 0 0 0 1 0
Flagellaria 0 1 0 0 0 ? 1 0 0 0 1 0 0 0 0 0 3 0 1 0 1 0 3 0 1 0 1 0 0 0 2 0 0 0 1 1 0 0 0 0
Geitonoplesium 0 1 0 0 0 ? 1 1 0 0 1 1 0 0 0 0 3 0 1 1 0 0 3 0 1 1 0 0 0 1 0 2 0 0 1 0 0 0 1 0
Joinvillea 0 1 0 0 0 ? 1 0 0 0 1 0 0 0 0 0 3 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0
Palisota 0 1 0 0 0 ? 1 1 0 0 1 1 0 0 0 0 2 1 1 0 1 0 0 — — — 0 — 0 0 2 0 0 0 1 1 0 1 0 0
Potamogeton 0 1 0 0 0 ? 1 0 0 1 1 0 0 1 0 0 2 0 0 0 0 0 2 0 0 0 0 0 0 0 1 0 0 0 1 1 0 1 0 3
Ruppia 0 1 0 0 0 ? 1 0 0 1 1 0 0 1 0 0 2 1 — 0 0 0 0 — — — — — 0 0 1 0 0 0 1 1 0 1 0 3
Smilax 1 1 0 0 0 ? 1 0 0 0 1 0 0 0 0 1 3 0 0 0 0 0 3 0 0 0 0 0 0 0 2 0 0 1 1 0 0 0 0 0
Sowerbaea 0 1 0 0 0 ? 1 1 0 0 1 1 0 0 0 0 3 0 1 0 1 0 0 — — — — — 0 0 0 0 0 0 1 0 0 ? ? 0
Tacca 0 1 0 2 0 ? 1 1 1 0 1 1 1 0 0 0 3 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0
Triglochin 0 1 0 0 0 1 1 0 0 1 1 0 0 1 0 0 * 0 1 0 0 0 * 0 1 0 0 0 0 0 1 0 0 0 1 0 0 1 0 0
Zannichellia 1 1 0 0 0 ? * 0 1 0 0 — — — 0 0 1 0 1 0 0 0 0 — — — 0 — 1 0 ? 0 0 0 1 0 * 1 0 3
Taxon 41–45 46–50 51–55 56–60 61–65 66–70 71–75 76–80

Aechmea 0 * ? ? ? ? 0 0 0 ? 0 * 1 1 0 0 0 ? 0 1 1 1 — — 0 1 0 ? ? 0 ? ? 0 ? ? ? 0 3 1 1
Chamaedorea 0 * ? ? ? ? * 0 0 0 0 0 0 1 0 0 1 1 1 1 0 0 2 — 0 0 0 ? ? 0 ? 0 1 ? ? ? 0 2 1 1
Cordyline 0 ? ? ? ? ? ? ? ? ? ? ? 0 1 ? 0 0 1 0 1 0 1 — — 0 1 ? 0 ? ? ? 0 0 ? ? ? ? 2 1 1
Dianella 0 0 ? 0 0 0 0 ? 3 ? 0 0 0 1 1 0 0 0 0 1 0 1 — — 0 1 * 0 ? 0 0 ? 0 0 1 1 1 3 1 1
N O V E M B E R 2015 , V O LU M E 102
Flagellaria 0 0 0 0 0 1 1 ? 4 ? 1 0 0 1 0 0 1 1 1 1 0 0 1 — 0 0 0 1 ? 0 1 ? 2 0 1 1 0 2 1 1
Geitonoplesium 0 ? ? 0 0 0 0 ? 0 1 ? 0 0 1 0 0 0 0 0 1 0 1 — — 0 1 1 0 ? 0 ? 0 0 ? ? ? ? 3 1 1
Joinvillea 0 0 0 0 0 1 1 1 4 0 1 1 0 1 0 0 1 1 0 1 0 0 1 — 0 0 0 1 ? 0 0 ? 2 0 1 1 0 2 1 0
Palisota 0 * ? 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 1 0 1 — — 0 0 0 ? ? 1 0 0 2 0 1 1 ? 3 1 1
Potamogeton ? 4 0 0 0 0 0 0 — 1 0 1 0 — * 0 0 1 0 0 0 0 0 — — 0 0 0 1 0 0 0 * 0 1 * 0 2 1 1
Ruppia * 3 0 0 0 0 0 0 6 0 0 1 0 — 0 0 0 — 0 0 0 0 1 — — 0 0 0 1 0 — 0 1 0 1 1 0 2 1 1
Smilax 0 * 0 0 0 0 0 ? * 0 ? 0 0 — 0 0 1 1 0 1 0 0 — — 0 0 0 ? ? 0 0 1 2 1 1 1 1 3 1 1
Sowerbaea 0 ? ? 0 0 0 0 ? 0 0 ? 0 0 1 1 0 0 1 0 1 0 1 — — 0 1 ? 0 1 0 ? 0 0 ? ? ? ? 5 0 0
Tacca 0 0 0 0 1 1 0 0 0 0 0 0 1 1 0 0 0 1 0 1 0 1 — — * 0 0 0 1 0 0 0 2 0 1 1 0 3 1 1
Triglochin 0 4 0 0 0 0 0 0 — 0 0 * 0 0 0 0 0 1 0 * 0 0 0 — — 0 0 0 1 0 0 0 0 0 1 1 0 2 1 1
Zannichellia 1 4 0 0 0 0 0 3 — ? 0 0 0 — 0 0 0 0 0 0 0 0 1 — — 0 0 0 1 0 — 0 0 0 1 1 1 2 1 1
Taxon 81–85 86-90 91–95 96–100 101–105 106–109

Aechmea 1 1 1 3 2 * 0 0 1 0 2 2 1 0 0 ? 3 0 2 ? 1 0 0 0 0 0 0 ? ?
Chamaedorea 1 0 1 1 2 1 1 1 1 0 1 2 0 0 0 ? * 1 1 — 0 0 0 0 0 0 0 2 1
Cordyline 1 0 1 0 2 1 0 0 1 0 1 1 2 0 0 ? ? ? ? ? ? ? ? ? 1 0 0 2 1
Dianella 1 1 1 0 1 1 0 0 0 0 2 1 1 0 0 0 ? 1 2 0 0 0 0 0 1 0 0 1 1
Flagellaria * 0 1 2 4 0 0 1 0 1 1 1 2 0 0 1 ? 1 ? ? 1 0 0 0 0 0 0 1 1
Geitonoplesium 1 1 1 0 1 0 0 0 1 0 2 1 0 1 0 ? ? 1 ? ? 0 0 0 0 1 0 0 2 1
Joinvillea * 1 1 0 1 2 0 1 0 0 1 3 0 0 0 ? 3 1 1 — 1 0 0 0 0 0 0 2 1
Palisota 1 1 2 0 1 1 0 0 1 0 2 1 0 1 0 ? ? ? 1 — 1 0 0 0 0 0 0 1 2
Potamogeton 1 0 1 2 0 0 1 0 0 0 3 4 2 0 0 0 1 1 2 ? 1 0 0 0 0 0 0 1 1
Ruppia 1 0 1 0 2 1 1 0 0 1 3 5 2 0 1 0 1 0 * ? 0 0 0 0 0 0 0 1 0
Smilax 1 1 1 2 0 1 0 0 1 1 1 0 0 0 0 0 ? 1 1 — 1 0 0 0 0 0 0 1 1
Sowerbaea 0 0 1 0 2 1 0 0 0 0 1 0 2 0 0 ? ? 1 ? ? 0 0 0 0 1 0 0 2 1
• T H A D E O E T A L. — A N ATO M Y O F F L E S H Y F R U I T S I N T H E M O N O COT S
Tacca 1 1 1 0 2 0 0 0 0 0 1 1 0 0 0 0 ? 1 1 — 0 0 0 0 0 0 0 1 1
Triglochin 1 0 1 0 1 0 0 0 0 0 2 1 2 0 0 0 1 0 1 — 0 0 0 0 0 0 0 1 ?
Zannichellia 1 0 1 0 0 0 0 0 0 0 2 3 2 0 0 1 1 0 2 ? 0 0 0 0 0 0 0 ? 0
• 1779

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RESEARCH ARTICLE

Anatomy of fleshy fruits in the monocots1

Taxon 41–45 46–50 51–55 56–60 61–65 66–70 71–75 76–80

Taxon 81–85 86-90 91–95 96–100 101–105 106–109

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