Leaf anatomy in the Asparagaceae as exemplified by Cordyline fruticosa, Dracaena reflexa,

& Sansiviera trifasciata

V. Casison and J. Elevazo

Students of Biology 101 Laboratory, Section LB1, Department of Biology, College of Arts and Sciences, University of the Philippines Manila,
Ermita, Manila 1000

Abstract. Plant anatomy has always been instrumental in providing means for taxonomically
classifying plants. Leaf anatomy in particular has proven to be useful in resolving species
circumscriptions in a few Asparagaceae taxa. In this study, leaf anatomy was described and
compared across 3 Asparaceae species of different genera: Cordyline fruticosa, Dracaena reflexa,
and Sansevieria trifasciata. Results showed variation in leaf anatomical features. Leaves differed
in stomata complex type and fiber bundle abundance and distribution. Unifying features include a
uniseriate epidermis, convergent mesophyll, and presence of raphide calcium oxalate crystals.
Sunken stomata and leaf depressions are notable xeromorphic characters. A simple investigation
of leaf anatomy supports the classification of the 3 species at the family level. Relationships
below the family level might not be manifested in leaf anatomy, therefore an extensive and
deeper anatomical examination is recommended.

Introduction Allium species determined that stomatal

Plant anatomy has been a long-standing tool
for plant taxonomy. In as early as the
timeslikes of: Theoprastus, (371-297 BC),)
dubbed as the “father of botany”,, who used
morphological charactersplant morphology
in determining the likeness of plant species
in his Historia Plantarum (Trinity College
Dublin, 2011); to the likes of Carl vonVon
Linne, who used microscopic evidencewith
his contribution to classify plants taxonomy
in his seminal workwork Species Plantarum
introducing the binomial nomenclature, who
used microscopy in specimens for the
aforementioned work in classifying plants
(Ford, 2009), plant anatomy has proven
itself as a usefuluniversal tool in for the
bases of taxonomy in history, and even to
this day with the advent of genetics,
cladistics, and phylogeny, plant anatomy
still holds sway in providing means for
simple differentiation between species.
Leaf, anatomy, as a subset of the huge
disciplinean extension of plant anatomy,
hashave also had its specific share of the
literature. For, for one, epidermal studies of
index and arrangement werewas diagnostic
of plant taxa (Tan & Lin, 2015). Other
studies onconcerning Ficus species’ leaves
as well as Malpighiaceae members
showspecies showed that an extensive study
of leaf mesophyll, trichomestrichome,
epidermal character, glandular placements,
and phloem positions, among others,
wereother things, was instrumental to
identification (Robbertse, Greuning, &
Grobbelaar, 1984), (Arau´jo, Ariste´a, Silva,
& Meira, 2010).
Anatomical studies ofFor the species under
Asparagaceae members are scarce and Formatted: Font: Not Italic
conducted individually as taxonomic
monographs of an entire genus.: Firstly,
studies on Cordyline were concerned mostly
on morphology (Fisher & Tomlinson, 1972),
and stem anatomy (Cattai & De Menezes,
2010). Secondly, microscopic leaf anatomy
was used to resolve 8 species of Dracaena
(not including D. reflexa). Notable finding
was the inter-specific variation in terms of
xeromorphic features, with all species
exhibiting isobilateral mesophyll and water-
storage cells (Klimko, Nowińska, Wilkin, &

1
Wiland-Szymańska, 2018). A similar study
on 49 Sansiviera taxa found variation in
xeromorphic charactersmosty characteristic
of xeromorphs that are useful in
classification (Koller & Rost, 1988). Despite
the demonstrated value of leaf anatomy in
taxonomy, to our knowledge no one has
attempted to study the leaf anatomical
variation across major subfamilies and
genera of Asparagaceae.
ThisThe study aimedaims to describe and
compareobtain anatomical
featuresdifferences of leaf samples across 3
Asparagaceae between the species:
Cordyline fruticosa (subfamily
Lomandroideae) commonly known as
cabbage treecordyline, Dracaena reflexa
(subfamily Nolinoideae) commonly known
as the dragon plant or pleomele, and&
Sansiviera trifasciata (subfamily
Nolinoideae), commonly known as snake
plantSansiviera, all belonging under the
Asparagaceae family. The study was done to
know the limits of leaf anatomy in
determining the taxonomic classifications of
plants. The plants were specifically chosen
due to their ubiquity as decorative plants
found within the University of the
Philippines Manila, and due to their
convenient relation to each other as, being
monocots under the Asparagaceae family.
The objectives of the study are as follows:
obtain leaf samples and slides of the species
so as to see their anatomical characters; and,
a comparative anatomy of the
aforementioned species to know the extent
of their differences.
Methodology
Leaf samples Specimens obtained were
sourced within theThe University of theThe
Philippines Manila. Identification was
facilitated by the distinct appearance of the
leaves, especially by the variegated pigment,
and was confirmed by a botany professor (F.
Evangelista, personal communication, May
18, 2019)., inclusive of: Cordyline fruticosa,
Dracaena reflexa, & Sansiviera trifasciata
leaf samples.
Freehand cross-sectionsMost of the study
was done with light microscopy, of which
cross-sectional slides of the leaf lamina,
petiole, and midrib werewas obtained,
whenever possible to study the internal leaf
organization. Epidermalthe best of the
researcher’s ability. Supplementing the fresh
slides, epidermal imprinting usingwas done
as well with the use of nail polish and
freehand paradermal sections were done as
well to studythe subsequent application light
microscopy of the epidermis and stomata.
Fresh sections were then subjected to bright-
field microscopy.obtained film.
Leaf
The anatomical features of importance,
specific to leaf anatomy included the
following: Epidermis and stomata,
Mesophyll type, crystals, fiber bundles,
vascular tissue arrangement, and the
existence of bundle caps.
Most of the microscopy was done on May 9
and May10, 2019 at Gusaling Andres
Bonifacio room 401 of The University of the
Philippines Manila.
Results and Discussion
Epidermis and Stomata
Epidermal cells proper are rectangular in
surface view with the longitudinal axes
oriented apical-basal (Figure 1), while in
transverse sections, the cells are isodiametric
and smaller (Figure 2, 3); this suggests a
flat,tabular 3D shape.
2
 these depressions may be an adaptation to
Resul arid conditions.
Sansiviera trifasciata 400x
Cordyline fruticosa 100x

Cordyline fruticosa 400x

Dracaena reflexa 100x

Figure 1. Paradermal sections of Sansiviera

trifasciata, & Cordyline fruticosa. Notice: the arrows
at Sansiviera, two subsidiary cells seen and reduced
bordering the guard cells (type II); arrows at
Cordyline, two subsidiary cells bordering the guard
cells (type III), but as opposed to Sansiviera not
reduced in size.
Figure 2. Petiolar sections of Cordyline fruticosa, &
Dracaena reflexa. Notice: petiolar sections of
Sansiviera trifasciata were not obtained due to the
succulent nature of sansiviera leaves, being consistent
at the blade, and near the node.

All leaf epidermises are uniseriate and Cordyline fruticosa 100x

glabrous lacking trichomes (Figure 3).
Stomatal distribution is amphistomatic for
all leaves as is typical of unifacial monocot
leaves (Evert, 2006). Sunken stomata are
also common to all 3, however those of
Cordyline and Dracaena are localized in
leaf depressions accentuating the sunken
position (Figure 2). As is known of the
function of sunken stomata (Evert, 2006),

3

Sansiviera trifasciata 40x
Dracaena reflexa
Green bands 100x
White bands 100x
Figure 3. Blade sections of Cordyline fruticosa,
Dracaena reflexa, & Sansiviera trifasciata. Notice
the thick mesophyll in Sansiviera causing its bundles
to form a non-linear arrangement. Also, Dracaena
reflexa exhibiting alternating bands of storage
mesophyll and chlorenchymatous mesophyll
Stomata complex type is perhaps the most
important diagnostic character. Guard cells
are kidney-shaped in all leaf samples and are
flanked by subsidiary cells (SC) (Figure
1). C. fruticosa exhibits Type III typical of
grasses which have 2 SC on each side of the
guard cells; D. reflexa exhibits Type I with 4
equal-sized SC; and S. trifasciata exhibits
Type II with 2 small and 2 large SC.
However, these stomata complex types are
likely species-specific in these taxa rather
than being a consistent feature of a genus
because inter-specific variation were noted
in anatomical studies of Sansevieria (Koller
& Rost, 1988) and Dracaena (Klimko et al,
2018).
Mesophyll
Mesophyll is convergent and compact in all
3 specimens. There is no differentiation into
a chlorenchymatous palisade and
aerenchymatous spongy in the lamina.
However, the distribution of chlorophyllous
cells might have taxonomic importance.
Cordyline chlorophyllous cells are
uniformly distributed in the lamina
mesophyll while the epidermal cells at the
peripheries have a violet pigment. Those of
Dracaena characteristically alternate with
achlorophyllous regions reflecting their
external pigment pattern of white-and-green
bands. Those of Sanevieria are localized at
the peripheries beneath the epidermis, much
of the mesophyll comprising
achlorophyllous storage parenchyma which
accounts for its succulence.
Cross-sections of the sheathing leaf bases of
Cordyline and Dracaena differ from lamina
sections which are flat. In contrast, leaf
bases have thicker mesophyll and have an
abundance of fiber bundles. The fiber
bundles of Cordyline seem to be scattered in
the mesophyll while those of Dracaena are
predominant in the adaxial side near the
4
upper epidermis. These fibers can be
considered xeromorphic features of the
Asparagaceae leaves (Blunden & Jewers,
1972; Koller & Rost, 1988; Klimko et al,
2018), and the thicker and more numerous
they are, the greater the degree of
xeromorphism. Although they do not
function directly to prevent water loss, their
presence might have allowed evolution of
large, broad, sometimes succulent, leaves as
supporting structures. It is surprising
therefore that these fiber bundles are absent
in S. trifasciata leaf samples. These may be
due to limitations of the freehand technique
employed because there is reason to expect
fiber bundles in the Sansevieria genus
(Koller & Rost, 1988).
The only observable ergastic substance are
raphide Calcium oxalate crystals which are
present in all leaf specimens. This might be
the most important feature that unifies not
only the 3 species under examination but
also the rest of Asparagaceae. Few other
monocots produce this crystal type in
abundance among which are members of the
arum family Araceae (Prychid & Rudall,
1999).
Vascular Tissues
The vascular bundles in the sheathing base
of Cordyline are scattered much like in the
stem (Figure 2). Those of Dracaena are
arranged in a single file whether at level of
the base or lamina. Sansevieria notably has
numerous smaller vascular bundles scattered
in the mesophyll. Because of its succulent
nature, base and lamina anatomy were
expected to be similar. All vascular bundles
are collateral with the phloem abaxial to the
xylem. Sclerenchyma fibers were also found
to cap the phloem.
Conclusions and Recommendations
Leaf anatomy is an important consideration
of taxonomic classification, as demonstrated
by this study and others. The numerous
characters of a leaf can help in diversifying
similarities and differences between plants,
which can help in their classifications, as
well as species identification.
Specifically for the species examined under
Asparagaceae, diagnostic features include
stomata complex type, mesophyll thickness,
and fiber bundle distribution. Unifying
features are those characteristic of monocot
leaves in general rather than Asparagaceae
such as amphistomatic distribution and
convergent mesophyll with no prominent
midvein. On the other hand, some features
less common in monocots but prevalent in
Asparagaceae include the sunken stomata
and raphide crystals. Thus, a simple
investigation of leaf anatomy supports the
classification of Cordyline fruticosa,
Dracaena reflexa, Sansevieria trifasciata at
least at the family level. At lower taxonomic
ranks, leaf anatomical features here
presented do not seem to support the closer
affinity of the Dracaena genus to
Sansevieria (both under subfamily
Nolinoideae) as determined by conventional
phylogenetic analysis. It is possible that
close relationships such as these are
manifested in other aspects of plant anatomy
(i.e., stem, root and floral) and not in the
leaves.
As a recommendation for future studies,
extensive anatomical techniques should be
performed. This must include staining to
better observe fundamental tissue placement
as well as to determine the existence of other
ergastic substances.
5

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