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Acua-C.
35(1):1-9.
& Marn-M.
2013
ABSTRACT
The genus Zamia is morphologically and ecologically the most diverse of the
Cycadales. Colombia is the country where most living species of Zamia have been
recorded. Even though in the past this genus extended beyond the Neotropical region,
nowadays it is almost entirely restricted to that biogeographical realm. As with the
remaining Cycadales, this genus shows several unusual anatomical traits in the
leaflets. The objective of this research was to study and compare the leaflet anatomy
of Zamia disodon and Z. restrepoi and establish possible relationships between the
anatomical traits and the habitats of these species. We found that both species share
several unusual traits with each other and with other species of the genus, such as
the parenchyma morphology, the distribution of tissues between the veins and the
stomata morphology. The main differences between these species were seen in their
fiber clusters and in the distribution and abundance of the photosynthetic tissue near
the vein areas. The presence of stomata both adaxially and abaxially could be the
result of the very wet habitats where both species are native, while the presence
of fiber clusters just at the vein areas could account for the longevity of the leaves
of Zamia. On the other hand the unique traits of Zamia restrepoi are related to the
presence of a midvein, a trait not found in any other species of Zamia.
RESUMEN
back in the Mesozoic (Norstog & Nicholls variation of external leaflet morphology
1998): a thick cuticle, a strongly developed of several West Indian species. Stevenson
hypodermis with thick walls and sunken (1981, 1990), Stevenson et al. (1996) and
stomata usually present only on the abaxial Norstog & Nicholls (1998) provided detailed
surface of the leaflets. All species of Zamia descriptions about the general foliar anatomy
have pinnate leaves (Jones 2002) but the and the differences between the genera of the
number of leaflets per leaf varies considerably order. Zuo et al. (2004) have even analyzed
between species. The main axis of the leaf how the ultrastructure and function of the
consists of the petiole, which can be prickly chloroplasts change relative to carbon
or smooth, and the rachis which supports dioxide concentrations. The scarcity of
the leaflets. The young leaves show erect recent literature on cycadalean anatomy
vernation in aerial stemmed species, and shows the need for more research, especially
inflexed vernation in those with subterranean in species that are virtually unstudied, as
stems (Stevenson 1981), probably to protect is the case for most Central and South
the fragile new leaves as they emerge from American species of Zamia. Anatomical
the soil. The internal anatomy of the petiole- traits can be helpful in trying to establish
rachis axis is characterized by an abundance the relationships of the species within the
of parenchyma along with mucilage channels genus, as has happened with Cycas, a genus
and relatively few and small discrete in which the series and species groups have
vascular strands, which show an omega similar foliar anatomy (Hill 1996). Previous
() shape, if seen in cross section (Norstog studies of Zamia fairchildiana L.D. Gmez
& Nicholls 1998). The leaflets are variable and Z. neurophyllidia D.W. Stev. from Costa
between species in regards to shape, length, Rica confirmed that even though the general
width, margin and texture, and do not show a anatomical features within the genus follow
distinct main midvein (except in one species, a common general pattern, there are many
Z. restrepoi). Internally, in some species at details which have potential taxonomic
least, such as Zamia integrifolia, the palisade applications, and also demonstrated many
parenchyma is indistinct, unlike in Cycas interesting features that could allow these
(Norstog & Nicholls 1998). However, there species to survive in their natural wet forest
is a subepidermal compact parenchyma under undergrowth environments (Acua-Castillo
the adaxial surface in some tropical species. y Marn-Mndez 2012).
Also, idioblasts (Norstog & Nicholls 1998,
Jones 2002) and occasionally isolated cells The objective of this research was to
(Acua-Castillo & Marn-Mndez 2012) study and compare the leaflet anatomy of
with thickened secondary walls can be found Zamia disodon D.W. Stev. & Sabato and
in the mesophyll. Z. restrepoi, both native to Colombia, and
establish possible relationships between the
Zamia foliar anatomy, both internal and anatomical traits and the habitats of these
external, has been studied in previous species.
works. Probably the most relevant one is
that by Greguss (1968), which considered MATERIALS AND METHODS
most cycad species known at the time,
and was centered mostly on characters We worked with Zamia disodon and Z.
useful in paleobotany, such as epidermal restrepoi. Both species inhabit wet forests
anatomy. More recent research includes to rain forests in the lowlands and mid
the works by Newell (1985, 1989) which elevations of northwestern Colombia. Zamia
emphasized inter- and intra-population disodon is known only from Antioquia (and
Leaflet anatomy of Zamia disodor and Z. restrepoi
The veins of Zamia disodon are raised In the vein area and under the epidermal cells
adaxially. On the abaxial surface, the flat veins there are clusters of fibers, both abaxially and
are clearly distinguished from the area between adaxially (Figs. 1d, 1c). These fiber clusters
the veins due to differences in cell shape (Fig. are restricted to the veins, vertically aligned
1c). Also on the abaxial side of the vein, some with the vascular tissues, and no similar
conic or truncated structures with a trichome- structures were seen between the veins. The
like appearance can be seen. These structures abaxial fiber cluster is trapezoid shaped and
are almost entirely restricted to the epidermis is notoriously more conspicuous than the
over the veins, although a few of them can be adaxial cluster, which is composed of rather
observed in areas between the veins (Fig. 1a). few cells. Between the fiber cluster and the
Leaflet anatomy of Zamia disodor and Z. restrepoi
vascular tissue there is parenchyma with Even though the stomata are more frequent
cells of variable diameter and relatively small in the areas between the veins, some can be
intercellular spaces. Occasionally, scarce found on the midrib edges. On the midrib it
thick walled cells can be seen. In the area is possible to see trichome-like projections
between veins it is possible to distinguish air over the surface. These structures are conical,
chambers (Fig. 1b). truncated or cylindrical (Fig. 2c). As in Z.
disodon, there are also similar structures in
The midrib of Zamia restrepoi is raised both the areas between veins, but in such areas
adaxially and abaxially, although it is more they are very rare.
prominent on the adaxial surface (Fig. 2b).
Acua-C. & Marn-M.
Under the adaxial epidermis of the midrib and Acua-Catillo & Marn-Mndez (2012)
there are 2-4 layers of fibers. Under these for other species of the genus: sunken guard
fibers there is spongy parenchyma, similar cells and raised subsidiary cells that form
to that observed in the area between the the walls of the epistomatic chamber. As
veins (Fig. 2b), formed by tubular cells, with pointed out by Norstog & Nicholls (1998),
rather large air chambers that are connected this is an adaptation for xeric environments,
to the spongy areas between the veins and in mesic habitat species, this could be
(Fig. 2a). Under the spongy parenchyma an evolutionary remnant that was present in
there is a relatively compact parenchyma the ancestors of the living species. As was
surrounding the vascular tissue. There are reported for Zamia fairchildiana and Z.
2-5 collateral vascular strands within the neurophyllidia by Acua-Castillo & Marn-
midrib and they are relatively small, with Mndez (2012), Z. disodon and Z. restrepoi
little xylem and crushed protophloem. The have trichome-like structures on their leaflets,
parenchyma towards the abaxial side, next especially on the veins. These seem to be the
to the vascular tissue, is more compact than basal cells of the branched trichomes, which
that under the adaxial surface, and with cells are very common in young leaves of Zamia
of larger diameters. Under it there are 3-5 (Stevenson 1981), although these are no
layers of abaxial, subepidermic fibers, even longer present on mature leaves. It appears
more developed than those located adaxially. that all that remains of the trichome is the
The fiber clusters are entirely restricted basal cell, after the branched apical cell has
to the veins, as in Zamia disodon. Rarely, been shed. This suggests that these trichomes
there are thick-walled cells dispersed in the are very fragile.
parenchyma (Fig. 2d).
In Zamia disodon, a very uncommon leaf
DISCUSSION feature was observed: the leaflet veins are
raised on the adaxial surface, but flat at the
Most cycad species are hypostomatic abaxial side of the leaflet. This is in contrast
(Greguss 1968), but in a few species, and to the general pattern, in which the leaflets
occasionally in some individuals of some are either flat or the veins are raised abaxially
species, it is possible to find amphistomatic (Jones 2002). However, the general structure
leaflets; however, the stomatal density on of the leaflet is very similar to that of other
the adaxial surface of the leaflet is usually species of Zamia, the main differences being
much lower than that of the abaxial surface. the texture of the leaflet and the size of the
This was noticed in the samples of Zamia fiber clusters above and below the vascular
disodon, in which the abaxial density of strands. These fiber clusters could indeed
stomata seemed higher compared to the have taxonomic importance. However, we
upper surface. It is likely that the density are not aware of their use in this way, even
of the stomata and their presence on just though in Cycas the location and abundance
the abaxial surface of the leaflet, or on both of foliar tissues have been applied in
surfaces, could be related to the atmospheric taxonomic studies (Hill 1993).
humidity and the climate, especially
because amphistomatic species are chiefly The conspicuous midrib seen in all leaflets
from very humid environments such as the of Zamia restrepoi was the main trait used
undergrowth of tropical wet forests (Greguss to separate this species in the genus Chigua
1968). The structure of the stomata and their D.W. Stev. (Stevenson 1990), although
subsidiary cells in the studied species is the the most recent genetic evidence suggests
same as that described by Greguss (1968) that what was considered Chigua is nested
Leaflet anatomy of Zamia disodor and Z. restrepoi
within a clade of mostly South American allow the leaflets to increase gas exchange
Zamia. Nonetheless, the foliar anatomy of in order to eliminate excess water and fix
Zamia restrepoi shows several unusual traits, carbon. In contrast to Cycas there is no
mostly related to the midrib. As was pointed evident transfusion tissue in Zamia (Norstog
out by Stevenson et al. (1996), the midrib in & Nicholls 1998), although in this genus
this species is very similar to that seen in the the elongated tubular spongy parenchyma
South African endemic, Stangeria eriopus cells could be involved in horizontal transfer
(Kunze) Baill., as it is composed mostly of of substances and are therefore probably
parenchyma and has several discrete vascular fulfilling the role of a transfusion tissue.
strands. However, in the samples analyzed we
found air chambers in the adaxial side of the ACKNOWLEDGEMENTS
midrib. This is a rare feature in leaf anatomy
and to our knowledge this is the first report We are indebted to Anders Lindstrm, from
of this characteristic in Cycadales. We think the Nong Nooch Tropical Botanical Garden,
that either the presence or size of such air Thailand, for providing the material for this
chambers is variable, and probably related study. To the staff of Centro de investigacion
to relatively higher transpiration rates. This en Estructuras Microscpicas at the
hypothesis remains to be tested. The spongy University of Costa Rica for their valuable
parenchyma of the vein is connected to the help during this research, especially to Ethel
spongy parenchyma of the blade. Besides the Snchez-Chacn and Alexander Rodrguez
particularities associated with such a midvein, for their help with the electron microscopy
Z. restrepoi has several traits in common with study and the image processing. We are
other species of Zamia. The main cluster of also thankful to Paul Hanson for reviewing
fibers is located on the abaxial side of the the manuscript. Financial support for this
leaflet, and even though those on the adaxial research was provided by Vicerrectora de
surface are abundant, they have thinner Investigacin, Universidad de Costa Rica
walls than the abaxial fibers. We suspect through grant No 810-B0-042.
that fiber location and development could be
related to structural factors adding resistance LITERATURE CITED
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(Cycadales). Caryologia 34(3): 255-260. Aceptado: 28/01/2013