American Journal of Plant Sciences, 2013, 4, 1863-1871
http://dx.doi.org/10.4236/ajps.2013.49229 Published Online September 2013 (http://www.scirp.org/journal/ajps)
Elaphoglossum (Dryopteridaceae-Fern) of Amazon
Rainforest in Brazil: Anatomic Characterization and
Adaptative Strategies*
Ana Carla Feio1#, Ana Cristina Andrade de Aguiar-Dias2, Raimunda Conceição de Vilhena Potiguara3†
1Doctoral Student in Botany, Universidade Federal de Viçosa—UFV, Viçosa, Brazil; 2Instituto de Ciências Biológicas, Universidade
Federal do Pará—UFPA, Belém, Brazil; 3Museu Paraense Emílio Goeldi—MPEG, Coordenação de Botânica, Belém, Brazil.
Email: #anacarlafeio@gmail.com
Received June 17th, 2013; revised July 17th, 2013; accepted August 20th, 2013
Copyright © 2013 Ana Carla Feio et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
ABSTRACT
This study describes the anatomy of sterile leaves of Elaphoglossum discolor (Kuhn) C. Chr., E. flaccidum (Fée) T.
Moore and E. laminarioides (Bory ex Fée) T. Moore, the most representative species of the genus in the Ecological
Park of Gunma in Pará State. It reports the main diagnostic characters and provides new systematic data fo r the group.
In addition, it locates the production and accumulation sites of bioactive compounds to determine possible adaptive
strategies of these species in the Amazon rainforest environment. Diagnostic structural features include stoma typology,
central veins and margin forms, type of mesophyll, an d the presence of schlerenchymatous sheaths in the cortex, among
others. Among the bioactive compounds related to defense adaptation are phenolic compounds, which occur in all three
species, and alkaloids and mucilage, which are exclusive to E. laminarioides. Of the three species studied, E. lami-
narioides has features that make it the best suited to the rainforest environment.
Keywords: Fern; Ombrophilous Forest; Epiphytes; Comparative Anatomy; Secondary Metabolites
1. Introduction
Because many of the features of Elaphoglossum Schott
ex Smith do not allow for the diagnosis of sterile indi-
viduals, it is one of the fern genera with the greatest need
for further studies [1,2]. In fact, most infrageneric classi-
fications are based on morphological characters such as
plant size, blade form and size, and scale color and type
[2-6]. However, most of these characters are only rele-
vant when plants are in their reproductive stage [3].
Elaphoglossum grows in primary formations and is
sensitive to environmental changes. Some species may
be resistant for some time in human modified habitats,
but different light and moisture conditions have consid-
erable influence on their morphology [7].
Since the 1970s, these species have been mentioned in
floristic studies in the Amazon [8-15]. Their constant
presence in inventories is due to different adaptation
strategies developed during their evolution, especially in
the Amazon rainforest environment. Such strategies in-
clude various phytochemical defenses, high resistance to
diseases in moist environments, high tolerance to acute
nutrient imbalance in substrates, plasticity to adapt to
different ecological opportunities, mycotrophic interact-
tions [16] and the epiphytic h abitat, in which Elaphoglos-
sum prevails [17].
The scarcity of studies on fern anatomy hinders the
understanding of their structural organization. Among the
main anatomical studies carried out on this genus are
[18], who analyzed rhizomes and leaves of Jamaican spe-
cies; [19] who described the leaf anatomy of Elaphoglos-
sum in Tucumán, Argentina and [20,21], who addressed
leaf architecture of Argentinean species by focusing on
their induments. None of these studies examined Brazil-
ian species, indicating the lack of the knowledge about
their anatomy and possible relationships to the environ-
ments where they grow.
The aim of this study is to characterize the anatomy of
sterile leaves of E. discolor, E. flaccidum and E. lami-
narioides, listing their main diagnostic characters and
providing new data on the systematics of this group, in
addition to identifying pro duction and accumulation sites
*Part of the dissertation of the first author.
#Corresponding author.
In memoria
m
.
Copyright © 2013 SciRes. AJPS
Elaphoglossum (Dryopteridaceae-Fern) of Amazon Rainforest in Brazil:
Anatomic Characterization and Adaptative Strategies
1864
of bioactive compounds, in order to determine possible
adaptation strategies of these species in the Amazon rain-
forest environment.
2. Material and Methods
2.1. Plant Materials
Botanical material was collected in the Ecological Park
of Gunma (PEG) located in Santa Bárbara do Pará
(01˚13'25''S , 48˚17'40''W), Pará State, Brazil. The area is
covered by lowland Amazon forest with a maximum
altitude of 100 m above sea level. Adult individuals with
totally unwound leaf blad es were collected. The vouchers
of the fertile material deposited in the herb ariu m MG are:
E. discolor (MG 195547; MG 193856), E. flaccidum
(MG 195545; MG 193858), and E. laminarioides (MG
195548; MG 195551; MG 195552 ; MG 193852).
2.2. Field Observations
Elaphoglossum discolor and E. flaccidum were always
found and collected in shady areas within the forest,
while E. laminarioides was found at the forest edges,
more exposed to the sunlight. Although the individuals of
the latter presented many leaves that seemed to have
been chewed, no visitors were observed during our ten
excursions to the field over two years.
2.3. Light Microscopy and Histochemical Tests
Leaves were fixed in FAA70 for 24 h [22], and in neutral
buffered formalin [23] and formalin-ferrous sulphate [22]
for 48 h. Leaf samples (n = 7) were dehydrated in a terti-
ary butanol series [22] and embedded in histological par-
affin. Leaves were cut into sections with a Leica RM
2245 microtome and stained in safranin and Astra blue
[24].
The following histochemical tests were applied by
hand to sections of the central vein: Sudan black B [25],
Nile blue sulphate [26], NADI reagent [27], copper ace-
tate/rubeanic acid [28], PAS reaction (Periodic Acid Schiff)
[29], ruthenium red [30], tannic acid/ferric chloride [31],
Lugol’s solution [22], ferric chloride [22], fluoroglucinol
[22], vanillin hydrochloric [32], Dragendorff’s [33] and
Wagner’s reagents [34]. Control sections without treat-
ment were also analyzed and photographed to compare
their color to that of the material fixed in FAA70 and
NBF. Photomicrographs were taken with a Zeiss Axiolab
microscope equipped with a Canon digital camera.
2.4. Scanning Electron Microscopy (SEM)
The samples fixed in FAA70 were dehydrated in an etha-
nol series [22], critical point dried with CO2 [35] and me-
tallized with gold for SEM analyses. Images were cap-
tured with an LEO 145 0 VP electron microscope. Frontal
views of epidermis were observed in a scanning electron
microscope because chemical and mechanical dissocia-
tion techniques were unsuccessful.
3. Results
3.1. Leaf Blade
In frontal view, the epidermis of each of the three species
presented sinuous, thin anticlinal walls on both faces
(Figures 1(a) and (b)), with subtle cuticular striations
(Figure 1(c)), as well as fungal hyphae along the surface.
All species were hypostomatic (Figures 1(a) and (b)).
Stomata are on the same level as the other epidermal
cells (Figures 3(a) and (e)). In E. discolor and E. flac-
cidum, stomata were exclusively polocytic (Figures 1(d)
and (e)), while E. laminarioides presented anomocytic
(Figure 1(g)) and copolocytic stomata (Figure 1(h)) as-
sociated to polocytic (Figure 1(f)) stomata. Cross sec-
tions of the epidermis of each species showed that it con-
sists of one layer of cells covered with a thin cuticle
(Figures 2(b), (f) and (j)). However, in E. laminarioides,
the outer periclinal walls are prominent on both faces
(Figure 2(j)).
In cross sections, the midrib differed according to spe-
cies: in E. discolor, both the adaxial and abaxial faces
(Figure 2(a)) are similarly concave; in E. flaccidum both
faces were concave, but the adaxial one was more pro-
nounced (Figure 2(e)) and in E. laminarioides, the ad-
axial face is flat (Figure 2(i)). The midrib also revealed a
pattern of the structural organization of the studied spe-
cies: schlerenchymatous cells subjacent to the epidermis
of both faces formed an arch comprised of 2 - 6 layers
(Figures 2(b), (f) and (j)) with the rest filled with paren-
chymatous tissue. The midrib enclosed two large and one
small vascular bundles, each of which was surrounded by
a cortical schlerenchymatous sheath (Figures 2(c), (g)
and (k)), which was thicker in E. laminarioi des . Adjacent
to this sheath was the endodermis, followed by a 2-lay-
ered pericycle (Figures 2(d) and (h)). Vascular bundles
were bicollateral, with curved xylem at their tips.
Mesophyll was homogenous (Figures 3(a), (e) and (i)),
with 4 - 6 strata of branched cells in E. flaccidum (Fig-
ures 3(e) and (f)) and lob ed cells in the oth er two species
(Figures 3(a), (b), (i) and (j)). In addition, the intracel-
lular spaces were conspicuous with scarce collateral bun-
dles.
Although these species had revolute leaf margins,
(Figures 3(c), (g) and (k)), they differed according to
taxa: in E. discolor, they were less revolute and the epi-
dermis had lignified walls (Figure 3(c)); in E. flaccidum,
they were rostriform (Figure 3(g)) and in E. laminario-
ides they were more revolute, with slightly truncated ex-
tremities (Figure 3(k)).
Copyright © 2013 SciRes. AJPS
Elaphoglossum (Dryopteridaceae-Fern) of Amazon Rainforest in Brazil:
Anatomic Characterization and Adaptative Strategies 1865
Figure 1. Scanning electron mic rographs of Elaphoglossum.
(a), (b) General view of the surface of E. laminarioides: (a)
Adaxial face, (b) Abaxial face, (c) Epidermis with striated
cuticle and fungal hyphae (arrow); (d)-(f) Details of polo-
cytic stomata: (d) E. discolor, (e) E. flaccidum, (f) E. lami-
narioides, (g) Anomocytic stoma of E. laminarioides, (h) Co-
polocytic stoma of E. laminarioides.
3.2. Petiole
Cross sections of the petiole were cylindrical (Figures
4(a), (e) and (i)), but concave on the adaxial surface of E.
laminarioides (Figure 4(i)). The epidermis of all species
was formed by thin-walled rounded cells (Figures 4(b),
(f) and (j)). The cortical region, which comprised 6 - 10
strata of schlerenchymatous cells, is more evident in E.
discolor and E. flaccidum. Below this strata, the rest of
the cortex was filled with parenchymatous tissue. Unlike
E. laminarioides (Figure 4(k)), the cortex sheaths of E.
discolor and E. flaccidum were inconspicuous (Figures
4(c) and (g)) in both the medial and basal regions of the
petiole. The petiole vascular system was similar to that
described for the midrib of the taxa, with the only differ-
ence being that it presented five vascular bundles at its
basal portion (Figures 4(d), (h) and (l)), which merged
Figure 2. Cross sections of the central vein of Elaphoglos-
sum. (a)-(d) E. discolor; (e)-(h) E. flaccidum; (i)-(l) E. la-
minarioides. (a), (e), (i) General view; (b), (f), (j) Detail of
the epidermal cells; (j) Cells with prominent outer pericli-
nal walls; (c), (g), (k) Vascular bundles; (d), (h) Detail of
schlerenchymatous sheath in the cortex and endodermis; (l)
Cells with granulose, compact content (asterisk). Bars: (a),
(e), (i) = 250 µm; (b), (c), (f), (g), (j), (k) = 50 µm; (d), (h), (l)
= 20 µm.
to three bundles as they approached the midrib median
region.
3.3. Secretory Tissues
In all the species analyzed, the secretory tissues were epi-
Copyright © 2013 SciRes. AJPS
Elaphoglossum (Dryopteridaceae-Fern) of Amazon Rainforest in Brazil:
Anatomic Characterization and Adaptative Strategies
1866
Figure 3. Sections of the leaf blade of Elaphoglossum. (a)-(d)
E. discolor; (e)-(h) E. flaccidum; (i)-(l) E. laminarioides. (a),
(e), (i) Cross sections of the blade showing homogeneous
mesophyll; (b), (f), (j) Longitudinal sections of the meso-
phyll; (b), (j) Mesophyll with lobed cells; (f) Mesophyll with
branched cells; (c), (g), (k) General view of the revolute
margins; (d), (h), (l) Detail of the cells with content; (l) Cell
with dense content (asterisk) and adjacent starch grains
(arrow). Bars: (a), (e), (i) = 50 µm; (b), (d), (f), (h), (j), (l) =
20 µm; (c), (g), (k) = 200 µm.
Figure 4. Cross sections of the petiole of Elaphoglossum.
(a)-(d) E. discolor; (e)-(h) E. flaccidum; (i)-(l) E. lamina-
rioides; (a), (e), (i) General view; (b), (f), (j) Detail of the
epidermis and adjacent schlerenchy matous cells; (c), (g), (k)
Detail of the vascular bundles in the median region; (d), (h),
(l) Vascular bundles in the basal region. Bars: (a), (e), (i) =
250 µm; (b)-(d), (f)-(h), (j)-(l) = 50 µm.
dermis (Figures 2(b), (f), (j), 3(e) and (i)), parenchyma
(Figures 3(d), (h) and (l)) (including that of the xylem
and phloem), and pericycle. No exudate was released
into the environment. Independent of the individuals or
parts, the exudate had a compact or granulose aspect, but
it may appear differently in neighboring cells (Figure
2(l)), or even in the same cell (Figure 3(l)). In the mate-
rial fixed in either FAA or NBF, exudate color varied
from yellow to light brown (Figures 5(a) and (b)). Exu-
date was more abundant on leaf blades and more con-
spicuous in E. laminarioides. The results of the histoche-
mical tests are shown in Table 1 and Figures 5(a)-(t).
Copyright © 2013 SciRes. AJPS
Elaphoglossum (Dryopteridaceae-Fern) of Amazon Rainforest in Brazil:
Anatomic Characterization and Adaptative Strategies
Copyright © 2013 SciRes. AJPS
1867
4. Discussion
The species analyzed showed significant variation in
their structural characters, especially leaf blades and his-
tochemical features. Based on our results, E. laminari-
oides was the most different in terms of structural varia-
tion and the highest number of bioactive compounds.
Some of the leaf structural features studied, such as sin-
uous epidermis and hypostomatic leaf blades, concur
with the descriptions by [18] for E. latifolium, E. musco-
sum, E. pallidum and E. villosum and by [20,21] for E.
crassipes (Hieron.) Diels, E. gayanum (Fée) T. Moore, E.
lindbergii (Mett. ex Kuhn) Rosenst., E. lorentzii (Hieron.)
H. Christ, E. pachydermum (Fée) T. Moore, E. pilosel-
loides (C. Presl) T. Moore and E. yungense De la Sota.
While polocytic stomata predominated in all species,
they were associated to copolocytic and anomocytic sto-
mata in Elaphoglossum laminarioides. Although polo-
cytic and copolocytic stomata are among the five types
that can be found in Elaphoglossum [36], anomocytic
stomata are not described for this genus. [37,38] did not
correlate this stoma type with those found in E. laminari-
oides, but Sen and De (1992) asserted that polocytic sto-
mata are never associated to anomocytic ones. Neverthe-
less, latter studies carried out by [20,21] identified ano-
mocytic stomata in Argentinean species of Elaphoglos-
sum. This variation in stomata typology is relevant since
it can be used as an infrageneric diagnostic feature.
As only E. flaccidum presented branched cells and ro-
striform margins, mesophyll and leaf margins are also
useful infrageneric features. [39] stated that homogene-
ous mesophyll with branched cells occurs only in Pteri-
daceae species that grow in shady habitats. Since this is
the case of E. flaccidum, the present study corroborates
their assertion.
One of the most common features mentioned in ana-
tomical studies on ferns is their lignified schlerenchyma-
tous sheath in the cortex [40-46], which only encloses the
petiole vascu lar bundles in E. laminarioides. It is consid-
ered a specialized character found in specimens of most
of the derived families, such as Dryopteridaceae [45].
Although no visitors were observed in loco, many
leaves were chewed and pathogens such as fungal hy-
phae were observed on leaf epidermis. This is likely due
to the presence of lipids, which are considered nutrition-
ally important for different classes of visitors and patho-
gens [47].
The starch grains observed on the leaf blade of the
studied species are believed to be transitorily stocked in
the chloroplast during the day and degraded at night to
maintain plant metabolism [48].
Although we know that epiphytes are more vulnerable
to water stress [49], E. laminarioides has always been
collected in areas more exposed to the sun. The muci-
Table 1. Results of histochemical tests on secreting of Elaphoglossum.
Species
Metabolites Tests
E. discolorE. flaccidum E. laminarioides
Total lipids Sudan black B +
Acids and neutral lipids Nile blue sulphate ++
Lipids
Free fatty acids Cooper acetate/rubeanic acid +
Terpenoids Essential oils and oil-resin NADI reagent
FFS + + ++
Total phenolic compounds Ferric chloride + + +
Tannins Vanillin hydrochloric
Phenolic compounds
Lignins Fluoroglucinol
Wagner’s reagent ++
Alkaloids
Dragendorff’s reagent ++
Starch Lugol’s solution
++ ++ ++
Mucilage Ruthenium red
Total polysaccharides PAS reaction ++
Polysaccharides
Mucilage Tannic acid/ferric chloride +
Notes: ++ = high positive; + = positive; = negative.
Elaphoglossum (Dryopteridaceae-Fern) of Amazon Rainforest in Brazil:
Anatomic Characterization and Adaptative Strategies
1868
Figure 5. Histochemical characterization of the secretion in leaf blade cross sections of E. laminarioides. (a), (b) Sections
without treatment, note yellow to light brown color; (c), (d) Detection of lipoic acid with Nile blue, note the secretion in the
cell (asterisk); (e), (f). Positive result for fatty acids with copper acetate/rubeanic acid. (g)-(j) Positive reaction for phenolic
compounds: (g), (h) Detection with FFS, note the secretion in the cell (asterisk), (i), (j) Detection with ferrous chloride; (k), (l)
Positive result for alkaloids: (k) Detection with Wagner’s reagent, (l) Detection with Dragendorff’s reagent; (m), (n) positive
reaction for starch grains; (o)-(r) Detection of polysaccharides with PAS reaction, note the secretion (asterisk) and starch
grains (arrow) in the cell; (s), (t) Positive reaction for mucilage with tannic acid/ferric chloride. Bars: (a), (c), (e), (g), (i), (k),
(m), (o), (s) = 150 µm; (b), (f), (j), (l), (q), (t) = 50 µm; (d), (h), (n), (p), (r) = 20 µm.
laginous compounds found exclusively in this species
thus serve to retain water and protect the plant against
herbivory [50], since some chewed leaves were observed.
When [51] studied fungus-insect-plant interactions, they
observed that insects feed on the fungal hyphae present
on the plants (mycophagy), which could explain the oc-
currence of both chewed leaves and fungal hyphae on
leaf surfaces.
Phenolic compounds are known components of defen-
sive adaptations. According to [52], they present pharma-
cological and antinutritional properties that inhibit lipid
oxidation and the proliferation of fungi. [53 ] asserted that
Copyright © 2013 SciRes. AJPS
Elaphoglossum (Dryopteridaceae-Fern) of Amazon Rainforest in Brazil:
Anatomic Characterization and Adaptative Strategies 1869
they are involved in adaptation processes as a defense
against radiation and pathogenic aggressions.
Likewise, alkaloids, found only in E. laminarioides,
are toxic and act as a defense against herbivores and pa-
rasites, in addition to playing an allelopathic role [54].
Although no Elaphoglossum monospecific formations
were observed in the field (which could lead us to induce
some kind of dominance), the potential allelopathic ac-
tivity of many fern genera has been proved, e.g. by [55]
for Pteridium Gled. ex Scop., and [56] for Dicranopteris
Bernh., Gleichenia Sm. and Sticherus C. Pr esl.
Thus, the structural data reported constitute new char-
acters of the representatives of Elaphoglossum in vege-
tative stage. The bioactive compounds identified in situ
also provide important unpublished data, adding infor-
mation on the species and suggesting the development of
adaptive strategies. This could be observed especially on
E. laminarioides, which in addition to the compounds of
communal occurrence on the species studied, was the
only species presenting alkaloids and mucilage. These
compounds possess peculiar properties related to herbi-
vore and pathogen avoidance, demonstrating that E. la-
minarioides is possibly better adapted to its environment,
producing secondary metabolites that are more varied
and directly related to adaptive success.
5. Acknowledgements
The authors thank Dr. Alejandra Vasco, New York Bo-
tanical Garden (NYBG), for information on the systema-
tics studies in the group and the National Coun cil for Sci-
entific and Technological Development (CNPq) for the
Master scholarship of the first author (CNPq 132809/
2009-5). We also thank Nicholas J. Walker and Anna
Luiza Ilkiu Borges Benkendorff for revising the English
of this manuscript.
REFERENCES
[1] R. E. G. Pichi Sermolli, “Adumbratio Florae Aetiopicae:
l5. Elaphoglossaceae,” Webbia, Vol. 23, No. 1, 1968, pp.
209-246. doi:10.1080/00837792.1968.10669885
[2] J. T. Mickel and L. Atehortúa, “Subdivision of the Genus
Elaphoglossum,” American Fern Journal, Vol. 70, No. 2,
1980, pp. 47-68. doi:10.2307/1546218
[3] J. T. Mickel and A. R. Smith, “The Pteridophytes of
Mexico,” Memories of the New York Botanical Garden,
Vol. 88, 2004, p. 282.
[4] A. Vasco, R. C. Moran and G. Rouhan, “Circumscription
and Phylogeny of the Elaphoglossum ciliatum Group (E.
sect. Lepidoglossa, Dryopteridaceae) Based on cpDNA
Sequences,” Taxon, Vol. 58, No. 3, 2009, pp. 825-834.
[5] A. F. Rojas-Alvarado, “New Taxa, New Records and
Redefined Concepts in the Elaphoglossum sect. Elaph-
oglossum subsec. Pachyglossa (Lomariopsidaceae) from
Mexico and Central America,” Revista de Biología Tro-
pical, Vol. 51, No. 1, 2003, pp. 1-32.
[6] A. F. Rojas-Alvarado, “A New Species in Elaphoglossum
sect. Elaphoglossum subsect. Pachyglossa (Dryopteri-
daceae) from Costa Rica and Panama,” Novon, Vol. 20,
2010, pp. 334-337. doi:10.3417/2008122
[7] A. C. Brade, “O Gênero Elaphoglossum (Pteridophyta)
no Brasil,” 2003. http://www.saude.unisinos.br/pteridos
[8] R. M. Tryon and D. S. Conant, “The Ferns of Brasilian
Amazonia,” Acta Amazononica, Vol. 5, No. 1, 1975, pp.
23-24.
[9] S. T. Rodrigues, S. S. Almeida, L. H. C. Andrade, I. C. L.
Barros and M. E. Van den Berg, “Composição Florística
e Abundância de Pteridófitas em três Ambientes da Bacia
do rio Guamá, Belém, Pará, Brasil,” Acta Amazonica, Vol.
34, No. 1, 2004, pp. 35-42.
doi:10.1590/S0044-59672004000100005
[10] J. Prado, “Flora da Reserva Ducke, Amazonas, Brasil:
Pteridophyta—Lomariopsidaceae,” Rodriguésia, Vol. 56,
No. 86, 2005, pp. 59-66.
[11] J. M. Costa, M. G. C. Souza and M. R. Pietrobom,
“Levantamento Florístico das Pteridófitas (Lycophyta e
Monilophyta) do Parque Ambiental de Belém (Belém,
Pará, Brasil),” Revista de Biologia Neotropical, Vol. 3,
No. 1, 2006, pp. 4-12.
[12] J. M. Costa and M. R. Pietrobom, “Pteridófitas (Lyco-
phyta e Monilophyta) da Ilha de Mosqueiro, Município de
Belém, Estado do Pará, Brasil,” Boletim do Museu Pa-
raense Emílio Goeldi Ciências Naturais, Vol. 2, No. 3,
2007, pp. 45-56.
[13] S. Maciel, M. G. C. Souza and M. R. Pietrobom, “Licó-
fitas e Monilófitas do Bosque Rodrigues Alves Jardim
Botânico da Amazônia, Município de Belém, Estado do
Pará, Brasil,” Boletim do Museu Paraense Emílio Goeldi
Ciências Naturais, Vol. 2, No. 2, 2007, pp. 69-83.
[14] M. R. P. Silva and S. M. Rosário, “Licófitas e Monilófitas
(Pteridophyta) da Floresta Nacional de Caxiuanã, Estado
do Pará, Brasil: Chave Para as Famílias e as Espécies de
Aspleniaceae e Blechnaceae,” Boletim do Museu Para-
ense Emílio Goeldi Ciências Naturais, Vol. 3, No. 2,
2008, pp. 151-163.
[15] J. M. Costa and M. R. Pietrobom, “Samambaias e Licó-
fitas do Parque Ecológico do Gunma, Município de Santa
Bárbara do Pará, Estado do Pará, Brasil,” Rodriguésia,
Vol. 61, No. 2, 2010, pp. 223-232.
[16] C. N. Page, “Ecological Strategies in Fern Evolution: A
Neopteridological Overview,” Review of Palaeobotany
and Palynology, Vol. 119, No. 1-2, 2002, pp. 1-33.
doi:10.1016/S0034-6667(01)00127-0
[17] H. Tuomisto and A. D. Poulsen, “Pteridophyte Diversity
and Species Composition in Four Amazonian Rain For-
ests,” Journal of Vegetable Science, Vol. 11, 2000, pp.
383-396. doi:10.2307/3236631
[18] P. R. Bell, “Studies in the Genus Elaphoglossum. III.
Anatomy of the Rhizome and Frond,” Annals of Botany,
Vol. 14, 1951, pp. 347-357.
[19] M. E. Guantay and M. H. Terán, “Estudio Morfoana-
Copyright © 2013 SciRes. AJPS
Elaphoglossum (Dryopteridaceae-Fern) of Amazon Rainforest in Brazil:
Anatomic Characterization and Adaptative Strategies
1870
tómico de las Especies del Género Elaphoglossum Schott
de Tucumán,” Lilloa, Vol. 38, 1995, pp. 73-84.
[20] M. C. Lavalle and M. Rodríguez, “Arquitectura Foliar y
Otros Caracteres Vegetativos en Especies Argentinas de
Elaphoglossum Schott ex Sm. (Lomariopsidaceae—
Pteridophyta),” Boletín de la Sociedad Argentina de Bo-
tánica, Vol. 42, 2007, p. 107.
[21] M. C. Lavalle and M. Rodríguez, “Indumento y Modelos
Epidérmicos en Especies Argentinas de Elaphoglossum
(Dryopteridaceae—Pteridophyta),” Boletín de la Socie-
dad Argentina de Botánica, Vol. 45, No. 3-4, 2010, pp.
245-256.
[22] D. A. Johansen, “Plant Microtechnique,” 2nd Edition, Mc-
Graw Hill, New York, 1940.
[23] R. D. Lillie, “Histopathologic Technic and Pratical Histo-
chemistry,” 3rd Edition, McGraw Hill, New York, 1965.
[24] D. Gerlach, “Botanische Mikrotechnik, Eine Einfuhrung,”
Georg Thieme, Stuttgart, 1969.
[25] A. G. E. Pearse, “Histochemistry Theoretical and Apl-
lies,” Vol. 2, C. Livingstone, Edinburgh, 1985.
[26] A. J. Cain, “The Use of Nile Blue in the Examination of
Lipids,” The Quarterly Journal of Microscopical Science,
Vol. 33, 1947, pp. 383-392.
[27] R. David and J. P. Carde, “Coloration Differentielle des
Inclusions Lipidique et Terpeniques des Pseudophylles du
Pin maritime au Moyen du Reactif Nadi,” Comptes Ren-
dus de lAcadémie des Sciences, Paris, Vol. 258, 1964,
pp. 1338-1340.
[28] P. Ganter and G. Jollés, “Histologie Normale et Patholo-
gique,” Vol. 1-2, Gauthier-Villars, Paris, 1969, 1970.
[29] W. A. Jensen, “Botanical Histochemistry: Principles and
Practice,” W. H. Freeman and Co., San Francisco, 1962.
[30] M. Gregory and P. Baas, “A Survey of Mucilage Cells in
Vegetative Organs of the Dicotyledons,” Israel Journal of
Botany, Vol. 38, 1989, pp. 125-174.
[31] P. Pizzolato and R. D. Lillie, “Mayer’s Tannic Acid-Fer-
ric Chloride Stain for Mucins,” Journal of Histochemistry
& Cytochemistry, Vol. 21, 1973, pp. 56-64.
doi:10.1177/21.1.56
[32] M. E. Mace and C. R. Howell, “Histochemistry and Iden-
tification of Condensed Tannin Precursors in Roots of
Cotton Seedlings,” Canadian Journal of Botany, Vol. 52,
1974, pp. 2423-2426. doi:10.1139/b74-314
[33] A. B. Svedsen and R. Verpoorte, “Chromatography of
Alkaloids,” Elsevier Scientific Publish Company, New
York, 1983.
[34] M. Furr and P. G. Mahlberg, “Histochemical Analyses of
Laticifers and Glandular Trichomes in Cannabis sativa,”
Journal of Natural Products, Vol. 44, No. 2, 1981, pp.
153-159. doi:10.1021/np50014a002
[35] J. J. Bozzola and L. D. Russel, “Eletron Microscopy:
Principles and Techniques for Biologists,” Jones and Bar-
lett Publishers, New York, 1991.
[36] U. Sen and B. De, “Structure and Ontogeny of Stomata in
Ferns,” Blumea, Vol. 37, No. 1, 1992, pp. 239-261.
[37] E. Fryns-Clae ssens and W. van Cotthem, “A New Classi-
fication of the Ontogenetic Types of Stomata,” Botanical
Review, Vol. 39, No. 1, 1973, pp. 71-131.
doi:10.1007/BF02860071
[38] W. R. J. Van Cotthem, “Stomatal Types and Systematic,”
In: J. A. Crabbe and B. A. Thomas, Eds., The Phylogeny
and Classification of the Ferns, Academic, Nueva York,
1973, pp. 59-71.
[39] D. Graçano, A. A. Azevedo and J. Prado, “Anatomia
Foliar das Espécies de Pteridaceae do Parque Estadual do
Rio Doce (PERD)-MG,” Revista Brasileira de Botanica,
Vol. 24, No. 3, 2001, pp. 333-347.
doi:10.1590/S0100-84042001000300012
[40] Y. Ogura, “Comparative Anatomy of Vegetative Organs
of the Pteridophytes,” 2nd Edition, Gebrüder Bornt Rae-
ger, Berlin, 1972.
[41] L. Atehortúa, “Contribuciones al Conocimiento de los He-
lechos del Gênero Elaphoglossum (Elaphoglossaceae),”
Actualidades Biológicas, Vol. 13, No. 50, 1985, pp. 83-
93.
[42] R. Bercu, “Anatomical Aspects of Asplenium adiantum-
nigrum L,” Journal of Biological Research, Vol. 2, 2004,
pp. 57-61.
[43] V. Hernández, T. Terrazas and G. Angeles, “Anatomía de
seis Espécies de Helechos del Gênero Dryopteris (Dryo-
pteridaceae) de México,” Revista de Biología Tropical,
Vol. 54, No. 4, 2006, pp. 1157-1169.
[44] A. Bagniewska-Zadworna and E. Zenkteler, “Ultra-
structure of Endodermis and Stele Cells of Dehydrated
Polypodium vulgare L. Rhizomes,” Acta Biologica Cra-
coviensia Series Botanica, Vol. 48, No. 2, 2006, pp. 73-
81.
[45] V. Hernández-Hernández, T. Terrazas and K. Mehltreter,
“Anatomía Vegetativa de Ctenitis melanosticta (Dry-
opteridaceae, Pteridophyta),” Boletín de la Sociedad
Botánica de México, Vol. 80, 2007, pp. 7-17.
[46] I. B. Suffredini, E. M. Bacchi and J. E. Kraus, “Estudo
Farmacognóstico Do Caule e Raízes de Microgramma
squamulosa (Kaulf.) Sota (Polypodiaceae),” Revista Bra-
sileira de Farmacognosia, Vol. 18, No. 2, 2008, pp. 279-
286. doi:10.1590/S0102-695X2008000200023
[47] H. G. Baker and I. Baker, “The Predictive Value of Nec-
tar Chemistry to the Recognition of Pollinator Type,” Is-
rael Journal of Botany, Vol. 39, No. 1-2, 1990, pp. 157-
166.
[48] P. L. Keeling and K. L. Myers, “Biochemistry and Ge-
netics of Starch Synthesis,” Annual Review of Food Sci-
ence Technology, Vol. 1, 2010, pp. 271-303.
doi:10.1146/annurev.food.102308.124214
[49] D. H. Benzing, “Vascular Epiphytes: General Biology
and Related Biota,” University Press, Cambridge, 1990.
doi:10.1017/CBO9780511525438
[50] A. Fahn, “Secretory Tissues in Plants,” Academic Press,
London, 1979.
[51] C. Gehring and A. Bennett, “Mycorrhizal Fungal-Plant-
Insect Interactions: The Importance of a Community Ap-
proach,” Environmental Entomology, Vol. 38, No. 1,
Copyright © 2013 SciRes. AJPS
Elaphoglossum (Dryopteridaceae-Fern) of Amazon Rainforest in Brazil:
Anatomic Characterization and Adaptative Strategies
Copyright © 2013 SciRes. AJPS
1871
2009, pp. 93-102. doi:10.1603/022.038.0111
[52] S. E. Soares, “Ácidos Fenólicos como Antioxidantes,”
Revista de Nutrição, Vol. 15, No. 1, 2002, pp. 71-81.
doi:10.1590/S1415-52732002000100008
[53] A. Farah and C. M. Donangelo, “Phenolic Compounds in
Coffee,” Brazilian Journal of Plant Physiology, Vol. 18,
No. 1, 2006, pp. 23-36.
doi:10.1590/S1677-04202006000100003
[54] T. Robinson, “Metabolism and Function of Alkaloids in
Plants,” Science, Vol. 184, No. 4135, 1974, pp. 430-435.
doi:10.1126/science.184.4135.430
[55] R. E. Stewart, “Allelopathic Potential of Western Bra-
ken,” Journal of Chemical Ecology, Vol, 1, No. 2, 1975,
pp. 161-169. doi:10.1007/BF00987865
[56] G. L. G. Soares and T. R. Vieira, “Inibição da Germi-
nação e do Crescimento Radicular de Alface (cv. ‘Grand
Rapids’) por Extratos Aquosos de Cinco Espécies de
Gleicheniaceae,” Floresta e Ambiente, Vol. 7, 2000, pp.
180-184.