International Research Journal of Plant Science (ISSN: 2141-5447) Vol. 4(6) pp.

168-191, June, 2013

Available online http://www.interesjournals.org/IRJPS
Copyright 2013 International Research Journals

Full Length Research Paper

Anatomical studies on the genus Euphorbia L. Saudi

Arabia (Subgenera: Triucalli, Ermophyton, Esula and
Chamaesyce)
Aldhebiani, A.1* and Jury, S.2
1

Department of Biology, King Abdulaziz University, Jeddah, Saudi Arabia.

2
Centre for Plant Diversity and Systematics, University of Reading.
*Corresponding Author Email: aaldhebiani@kau.edu.sa
Abstract

The genus Euphorbia is the largest in Saudi Arabia, even though no anatomical study has been done
intensively. In this study the epidermis, the stomata and the venation patterns have been investigated.
The shape of the epidermal cell in Euphorbia species in Saudi Arabia varies: polygonal, rectangular,
undulate or elongated. Moreover, the cell shape relies on the cell location on a leaf, i.e. the middle
region, the margin, the apex or above the vein. Furthermore, in some cases both leaf surfaces have the
same cell shape but more often they are unlike. Hairs are generally simple, unbranched and with a
warty ornamentation on their surface. Papillae occur only in one species E. hypericifolia .The most
common stomata type is anomocytic, while the rare type is actinocytic, recorded only in E. helioscopia.
Stomata of more than one type (have been encountered on the same leaf surface as in E. scordiifolia
and E. hirta. Venation patterns vary from one-veined, three-veined to those with four or more veins.
Keywords: Euphorbia, Saudi Arabia, stomata, venation, epidermal cells, anatomical characters.

INTRODUCTION
Euphorbiaceae, the spurge family, one of the major
flowering plant families: with 334 genera grouped in 52
tribes and 5 subfamilies, is considered as the sixth
largest family of Angiospermae. In Saudi Arabia,
Euphorbiaceae is represented by 15 genera (Andrachne
L., Flueggea Willd, Phyllanthus L., Clutia L., Chrozophora
Neck. ex Juss., Ricinus L., Mercurialis L., Erythrococca
L., Micrococca Benth., Acalypha L., Tragia L.,
Dalechampia L., Jatropha L., Croton L. and Euphorbia L.)
(Chaudhary, 2001). Among them, Euphorbia is the
largest and varies from herbs to shrubs and trees, and
from succulent to non-succulent plants. Species are
scattered all over the country but the succulent taxa
mostly occur in the South West Region. According to
Govaerts et al., (2000), the genus Euphorbia L. is the
third largest genus in the flowering plants (after
Astragalus [Fabaceae] and Psychotria [Rubiaceae]) with
about 2000 species distributed worldwide, both in Old
and New Worlds, and mainly in the tropical, subtropical
and warm temperate regions. In addition to the wide
distributional range, Euphorbia L. has various life forms,
as annual or perennial herbs, shrubs or trees. The

species can be prostrate or erect, monoecious or

dioecious and succulent or non-succulent. Most of the
succulent Euphorbia species are endemic to Africa.
Even though the genus Euphorbia is the largest in
Saudi Arabia, no extensive study on the anatomy of the
genus has been done.
This work has been done to revise the genus
Euphorbia in Saudi Arabia in three parts: morophological,
anatomical
and
phytochemical
studies.
The
phytochemistry study has been published in a separate
paper.
In the Euphorbiaceae, anatomical characters have
been found significant since the work of Pax, (1884). In
his work, Pax found the importance of laticifers as a
character to define natural groups. In addition, he used
the type of laticifers (articulate or non-articulate), phloem
characters and trichome type as the major anatomical
features to support his redivison of the major
suprageneric taxa in the family. The first significant
anatomical survey of the family was provided by
Solereder, (1899) who revised the groups of Radlkofer,
(1870) and added many observations by himself. Then,

Aldhebiani and Jury 169

Gaucher, (1902) tried to omit all the work of the German

school by publishing an independent anatomical survey
of the family. His work was criticized by Solereder,
(1908). Yet, it was valuable in providing a comparative
review of anatomical characters which was arranged by
tribe. Later, in a series of studies, Mahlberg, (1973; 1974;
1975; 1982) and Mahlberg et al., (1987) showed the
systematic importance of laticifers in Euphorbia,
especially with regard to the starch grains produced.
Furthermore, the vascular anatomy of petioles has been
studied in cross section by Dehay, (1935) who found a
considerable variation in stellar configurations. However,
it seems unlikely that this feature will be of general value.
Meanwhile, Miller and Webster, (1962) have used
differences in petiolar steles to separate Cridoscolus from
Jatropha. Then, Dehgan, (1982) could prove that the
petiolar stellar dissection were significant at the sectional
and sub-sectional level in some genera, such as
Jatropha. However, in a considerable number of taxa,
stipules have become reduced or are early deciduous.
For example, many species lack stipules, whereas they
are large and remarkable in others. In Euphorbia, the
presence or absence of stipules is a diagnostic character
for some sections and subgenera (Webster, 1994a).
Anatomical Characters in Euphorbia
Latex
The white latex is a useful distinguishing character in the
genus Euphorbia L. It is distributed throughout the plant
in a series of tubes derived from either single cells (nonarticulated laticifers) or articulated laticifers formed by the
fusion of several cells. The value of laticifer types as a
taxonomic marker in systematic comparisons between
and within families has been established by Carlquist,
(1961).The laticifer system of the mature Euphorbia plant
was explained by Gaucher, (1898; 1902). In addition,
Rosowski, (1968) studied the branched non-articulated
laticifers system in mature tissue of the internode and
node in transition from the node to and throughout the
mature leaf of E.supina Raf. He found that the latex
system in the E. supina stem is restricted to the cortex
and does not break through the leaf gaps. In the leaf
lamina he found that the widest laticifers are in
association with the vascular system. These laticifers
begin to branch from the base of the leaf and continue
throughout certain areas in the mesophyll. Moreover,
they are associated with the phloem and may send some
branches at right angles to the vein and between the
bundle-sheath cells.
Epidermis
Epidermal cells may vary greatly in size, shape and

outline from species to species, especially when seen

in a surface view. Generally, in dicotyledons, epidermal
cells have irregular shapes and sizes. Sometimes the
shape of the cells in both leaf surfaces are similar, but
more often they are different. The costal cells usually
differ from those intercostal regions; they tend to be
elongated in the direction of the veins. Some marginal
cells develop unicellular or multicellular prickles. Moreover,
anticlinal walls can be either very thin and hardly visible from
the surface or they may range through degrees of thickness
to be very thick (Kakkar and Paliwal, 1972).
One of the important epidermal features for all
taxonomists is the hairs. They can be glandular or nonglandular; and can be divided depending on the
component number of cells and degree of branching. The
occurrence of distinctive types of hair can be a valuable
character to recognize a whole family. Moreover, hairs
are more useful in determination at the level of genus or
species. In other words, variations in size and density
should be accepted in the differentiation of closely related
genera or species after a comprehensive investigation of
a wide range of material (Metcalfe and Chalk, 1950).
In a study of 150 species of Euphorbia, Kakkar and
Paliwal (1974) described the epidermal cells in Euphorbia
as circular, trapezoidal, rectangular or polygonal in
outline. Additionally, they found that the shape may vary
depending upon the location of the epidermal cell on the
leaf, i.e. the middle region, margin, apex or above the
vein. Some xeromorphic species exhibit a waxy covering
which takes on many crystalline forms. Hairs were also
recorded in the epidermis of Euphorbia in some species.
In addition, the papillae occurred in the surface view as
rounded structures in the centre of cell lumen.
Stomata
A stoma consists of the stomatal aperture and the pair of
guard cells that form it. Stomata usually tend to be on the
lower surface only. But in some cases this distribution
varies from species to species and depends on whether
the plant is a xerophyte or a mesophyte. They might be
superficial or sunken. Stomata sometimes are
surrounded by specialized epidermal cells which are
called subsidiary cells. These subsidiaries differ from
unmodified epidermal cells in shape, size and staining
properties (Baranova, 1992; Metcalfe and& Chalk, 1950;
Stace, 1965). On the other hand, the arrangement of
subsidiary cells, where present, is of the greatest interest
to the taxonomist. This variation is used to define the
different types of stomata. Occasionally species have
several types of stomata on one leaf, while some have
only one type for the species (Stace, 1984). In addition,
Van Cotthem, (1973) pointed out that those
morphological stomata types can provide not only
diagnostic characters but also very valuable taxonomic
ones or even phylogenetic clues. Metcalfe and Chalk,

170 Int. Res. J. Plant Sci.

(1950) had established some terms to replace the

representative family name proposed by Vesque,
(1889).
Anomocytic
was
substituted
for
the
ranunculaceous type; anisocytic replaced cruciferous,
diacytic the caryophyllaceous and finally paracytic for the
rubiaceous. The tetracytic type which can be found in
most of the monocotyledons was added by Metcalfe,
(1960). Later, Stace, (1965) proposed the term cyclocytic
for the narrow ring of four or more subsidiary cells
surrounding the stomata. Metcalfe and Chalk, (1950)
have named and defined the actinocytic type as stomata
surrounded by a circle of radiating cells. Three more
types were introduced by Van Cotthem, (1970),
hexacytic, epicytic and hemiparacytic. And some
intermediate types were added by Payne, (1970) who
described the helicocytic and allelocytic types in relation
to mesogenous forms of anisocytic, paracytic and diacytic
patterns. Stace, (1989) lists 35 types of stomata in
vascular plants. Closely related families are distinguished
by the presence of a specific type of stomata; such as
Acanthaceae and Scrophulariaceae separated by the
presence of diacytic stomata in the former as against
anomocytic in the latter. Moreover, some stomatal types are
distinctive of certain families: for example, Ranuculaceae
has the anomocytic type, Brassicaceae the anisocytic,
Caryophyllaceae diacytic, Rubiaceae paracytic and finally
Poaceae has the graminaceous type (Singh, 2004).
According to Metcalfe and Chalk (1950), the mature
stomata of Euphorbiaceae, are anomocytic, paracytic and
anisocytic. They are usually confined to the lower leaf
surface, more rarely on both surfaces of the lamina.
Paracytic stomata were reported by Tognini, (1897)
which are mesogenous in development in E. variegata
and Ricinus communis. On the other hand, according to
Raju and Rao (1977), stomata in the Euphorbiaceae
show considerable variation. They found that the woody
taxa have predominantly the paracytic stomata type,
while the anisocytic stomata are characteristic of the
herbaceous Phyllanthoideae. Moreover, they indicated
that Chamaesyce has a high percentage of anomocytic
stomata (Raju and Rao, 1987). Finally, a considerable
diversity of stomatal types were found in Euphorbia by
Kakkar and Paliwal, (1974). They reported that the
common type of stomata in Euphorbia species is the
anomocytic, even though stomata of paracytic and
anisocytic have been also observed.
Venation
Some features that give the leaf its structure are, for
instance, leaf shape, margin type and venation patterns.
These together form the leaf architecture as discussed by
Hickey, (1979). A more sophisticated knowledge of leaf
architecture has allowed a start in discriminating
phylogenetic trends from leaves (Hickey, 1973; RothNebelsick et al., 2001). Due to its importance, especially
for systematic classification, attention has been paid

largely to the architectural properties of leaf venation

(Khler, 1993). The careful description of venation in
cooperation with studies of other leaf anatomy details can
provide valuable taxonomic evidence. The works of Pray
(1955a; 1955b) suggest that the venation can be useful in
a comparative analysis. Moreover, Wagner (1979) has
proved that the vein patterns were significant for
classification at the level of species, genus and family in
ferns. Of course, this is more noticed and observed in
angiosperms especially dicotyledons where the
hierarchical network patterns of the veins are much more
advanced and complicated.
Leaf venation has been neglected for a long time in
the Euphorbiaceae until the work of Levin, (1986a;
1986b; 1986c). Levin developed a new insight into
relationships within subfamily Phyllanthoideae. No
detailed studies have been provided for other
subfamilies, except the work of Sehgal and Paliwal
(1974) on the tribe Euphorbieae. In part II of their study
on the leaf anatomy of Euphorbia, Sehgal and Paliwal
designated the venation in petiolate leaves as uni-, biand tri-veined according to the number of strands
entering the petiole or lamina base. They noticed that in
the 150 species under investigation, the majority bear
leaves belonging to the tri-veined category. Moreover,
they divided the latter into ornamented and
unornamented veins. When the veins are surrounded by
a parenchymatous sheath it is called ornamented. And
when this sheath is absent it is unornamented. Besides,
sometimes trachedial nodules are encountered at the
apices and in the serrations of the leaf. Small veins
usually form networks in the lamina. These networks
may vary in size and shape, and subdivide the area of
the mesophyll. The thinnest branches of the bundles
bounded by the smallest areas or regions are called
areoles. These areoles usually contain blind vein
endings. The degree of branching of these vein endings
varies in the leaves of different species. Therefore, this
can be a useful identification tool.
MATERIAL AND METHODS
Herbarium specimens were obtained from Kew (Kew
Garden herbarium, UK) and RIY (National Herbarium in
Riyadh). Since leaves are early deciduous in succulent
species, most of the herbarium sheets or spirit collections
of these species have no leaves. As a result, these
species were omitted from this anatomy study, except for
the stick-like Euphorbias: E. balsamifera subsp.
adenensis, E. schimperi, E. aff. schimperi, E. aff.
consobrina and E. cuneata. Only mature leaves were
used for studying the epidermis, stomata and venation.
Epidermal peel
This method was used to observe stomata, epidermal

Aldhebiani and Jury 171

cells and hairs.

1Leaves were rehydrated using polyoxyethylene
sorbitan monolaurate (Tween20) in water and heated for
30 minutes. Then, washed by water and conserved in
70% ethanol for future work.
2Both surfaces of the leaf were scraped and
peeled off under a dissection microscope with the aid of
fine forceps and a razor blade to remove loose cells.
3Samples were bleached by Jefferys Solution or
the bleaching agent Vortex for 10-20 minutes depending
on leaf thickness. Sometimes this was overnight.
4Samples were washed, mounted and observed
under a light microscope.
5Photographs were taken using a Leitz Diaphlan
polarizing microscope or a Reichert Polyvar 2
Microscope.
Staining by alucin blue was tried, but without
success. Samples became too dense or obscure due to
the presence of tannins and other compounds in the
herbarium specimens. Therefore, some samples were left
without staining.

l)
Mounted in 100% glycerol and sealed with Canda
balsam.
m)
Observed under light microscope.
n)
Photographs were taken using a Leitz Diaphlan
polarizing microscope or Reichert Polyvar 2 Microscope.
Scanning Electron Microscopy
1Dry specimens were mounted surface up on
scanning electron microscope stubs using Bostik No.1
adhesive.
2Stubs were sputter coated for 2-3 minutes with a
gold palladium alloy using an Edwards sputter coater to
give a coating about 15-20 nm thick.
3Samples were examined in FEI Quanta FEG 600
Environmental Scanning Electron Microscope (ESEM) in
high vacuum SEM mode.
4Photographs were taken using the computerized
digital system of the microscope.
RESULTS

Leaf clearing
Epidermis
The following method was adopted from Radford et al.,
(1974), with some modifications for use on herbarium
materials.
1The leaf was placed in a Petri dish and covered
with 5% Sodium Hydroxide (NaOH). The dish was
wrapped in cling film leaving a small gap in one area for
ventilation.
2This was microwaved for five seconds on
medium power.
3The Petri dishes were left on a hot plate at 35-37
degrees centigrade for a week or until the leaf was
transparent. The NaOH was changed twice a day for
each sample.
4When the leaf had cleared sufficiently, it was
washed in water and covered in 90% bleaching agent
Vortex for ten minutes to one hour depending on the
reaction of the sample.
5The leaf was washed again in water.
6Samples were dehydrated in ethanol series and
stained as following:
a)
30% ethanol
2 minutes.
b)
50% ethanol
2 minutes.
c)
1% safranin in 50% ethanol
4 minutes.
d)
Wash in water three times
e)
30% ethanol
2 minutes repeated 3 times.
f)
50% ethanol
2 minutes repeated twice.
g)
70% ethanol
2 minutes.
h)
90% ethanol
2 minutes.
i)
100% ethanol 2 minutes.
j)
Absolute ethanol: histoclear 1:1 2 minutes.
k)
Histoclear
2 minutes.

Results of the epidermal characters in Euphorbia species

under investigation are summarized in Table 1.
Cell shape
Depending upon the shape of the cell wall and
considering the results from Kakkar and Paliwal, (1974),
the shape of the epidermal cell varies: polygonal,
rectangular, undulate or elongated. Moreover, the cell
shape relies on the cell location on a leaf, i.e. the middle
region, the margin, the apex or above the vein.
Furthermore, in some cases both leaf surfaces have the
same cell shape but more often they are unlike. This
combination of cell shape characters gives each species
its unique leaf surface appearance.
The normal straight cell walls in both adaxial and
abaxial surfaces are shown in the following taxa: E.
schimperi, E. cuneata, E. helioscopia, E. dracunculoides
and E. balsamifera subsp. adenensis see Figure 1. In
addition, E. aff. schimperi and E. aff. consobrina have
also the same straight cell wall on both surfaces.
Meanwhile, E. grossheimii, E. retusa and E. pirottae have
elongated cells on both leaf surfaces (Figure 2 a and b).
On the other hand, the plicate cell walls on both adaxial
and abaxial surfaces occur in E. chamaepeplus, E.
hypericifolia and E. peplus (Figure 2 c and d), but the
walls are more folded on the lower surface. In some
species, the cell wall on the adaxial surface is straight,
whilst it is plicate on the abaxial such as E. hirta, E.

172 Int. Res. J. Plant Sci.

Table 1. Epidermal characters of Euphorbia taxa in Saudi Arabia.

Cell shape (cell wall)

Straight
Subgenus
Tirucalli
Ermophyton

Species
E. balsamifera subsp.adenensis

Ad.*

Ab.*

Ad.

Ab.

Plicate
Ad.

Ab.

Glabrous

Unicellular

Multicellular

Ad.

Ad.

Ab.
+

E.aff. schimperi

E. aff. consobrina

E. cuneata

E. acalyphoides

E. pirottae
E. helioscopia

+
+

Esula

E. retusa

E. grossheimii
+

Papillae

Ab.

E. schimperi

E. dracunculoides

E. falcate

E. peplus

Chamaesyce
*Ab.: abaxial
*Ad. : adaxial

Elongated

Hairs

++

E. chamaepeplus
E. schimperiana
E. Arabica

+
+

++
+
+

+
+
+

E. serpens

E. scordiifolia

E. granulate

E. inaequilatera

E. hirta

E. hypericifolia

++

+ : Present
++ : the plicate folded wall is morefully developed in the lower surface than in the upper one
+? : hardly observable by LM only

+?

+
+

+
+

Aldhebiani and Jury 173

Adaxial

Abaxial

C
E

D
Figure 1. Normal irregular epidermal cells in both adaxial and abaxial surfaces of the leaf (LM).
(A) E. balsamifera subsp. adenensis, (B) E. schimperi, (C) E. cuneata, (D) E. falcata, (E) Abaxial in E. dracunculoides

174 Int. Res. J. Plant Sci.

Adaxial

Abaxial

D
Figure 2. Error! No text of specified style in document. Both adaxial and abaxial surfaces
have the same shape of epidermal cells (LM). Elongated in A &B; plicate in C&D. (A) E.
grossheimii, (B) E. pirottae, (C) E. chamaepeplus and (D) E. peplus.

schimperiana, E. helioscopia, E. inaequilatera, E.

arabica, E. granulata, E. serpens and E. scordiifolia see
Figure 3 and Figure 4. This last group of species belong
to subgenus Chamaesyce except for E. helioscopia and
E. schimperiana.

Hairs
Hairs on the leaf epidermis can be both unicellular and

multicellular and may appear only on one surface or both.

Furthermore, hairs in the Euphorbia species under
investigation are generally simple, unbranched and with a
warty ornamentation on their surface. Papillae appear in
ESEM as outgrowths, whereas in the surface view they
give the impression of being rounded structures at the
centre of the cell. Prominent papillae occur only in one
species of Euphorbia in Saudi Arabia; this species is E.
hypericifolia (Figure 5 and 6), whereas in E. arabica they
were poorly distinguishable using LM.

Aldhebiani and Jury 175

Adaxial

Abaxial

D
Figure 3. Epidermal cells vary on leaf surfaces: adaxial surface has irregular cells while
the abaxial surface has plicate (LM).
(A)
E. schimperiana, (B) E. arabica, (C) E. serpens, (D) E. scordiifolia.

176 Int. Res. J. Plant Sci.

Adaxial

Abaxial

Figure 4. Epidermal cell shape varies on the adaxial and

the abaxial surfaces, straight cell walls in the former
while folded plicate walls occur on the latter (LM). (A) E.
granulata, (B) E. inaequilatera and (C) E. helioscopia.

Hemiparacytic stomata
on both surfaces

Figure 5. E. hypericifolia (LM).

Above photo: the adaxial surface with irregular epidermal cells and
very clear papillae as a circle in the middle of the cell.
Lower photo: the abaxial surface with plicate cells and papillae, a
hair base is obvious in the right side of the picture.

Aldhebiani and Jury 177

Adaxial

Abaxial

Papillae

Multicellular hairs in abaxial surface.

Figure 6. E. hypericifolia (ESEM), papillae on the adaxial and abaxial surfaces,

multicellular hairs are on the abaxial surface of the leaf only.

Figure7. Multicellular hairs on adaxial and abaxial surfaces of E. hirta.

178 Int. Res. J. Plant Sci.

Figure 8. Hairs, wart ornamentation on hair surfaces of (A) E. scordiifolia and (B) E. pirottae; (C) Short hairs on
abaxial surface of E. balsamifera subsp. adenensis.

In E. balsamifera subsp. adenensis, the hairs are short

and unicellular (Figure 8C) and they are located mostly at
the leaf margins and the mid-vein area on the abaxial
surface only.
Hairs on the leaf of E. pirottae are present on the
abaxial surface only and are both unicellular and
multicellular. Euphorbia granulata is similar to E. pirottae
in having both unicellular and multicellular hairs, but here
they occur on both surfaces of the leaf (Figure 8B).
On the other hand, in E. scordiifolia, E. hirta and E.
hypericifolia the hairs are multicellular and on both
surfaces in the first two species and on the abaxial only in
the latter (Figure 7). Moreover, E. hypericifolia and E.
hirta have long hairs when compared with the other
species (Figure 6).
No hairs were observed and leaves were glabrous in
the following species: E. serpens, E. arabica, E. peplus,
E. inaequilatera, E. falcata, E. helioscopia, E. cuneata, E.
retusa, E. grossheimii, E. schimperiana, E. schimperi,
E. chamaepeplus and the new variety of E. granulata.

Wax
The cuticle layer and wax in the Euphorbia species under
investigation vary from smooth to densely cover in wax
scales. This may be a thick smooth layer of wax which
sometimes has upright scales. In some species, only the
cell surface has a smooth wax layer whereas the wall
between cells has upright scales of wax as in E.
schimperiana, especially around the stomata pores
(Figure 9). In contrast, the anticlinal walls of some
species have a smooth layer of wax, whereas the cell
surface is densely covered with upright wax flakes or with
some stellate wax flakes in other species.
Stomata
Stomata were observed through LM and ESEM. In the
species under investigation, stomata commonly appear
on both leaf surfaces (amphistomatic leaves) except
in E. cuneata where the stomata are located only on the

Aldhebiani and Jury 179

E
Figure 9. Wax formation in Euphorbia species in Saudi Arabia: (A+B) smooth and
thick in E. peplus and E. schimperi; (C+D) smooth on top of cell while forming
upright scale flakes in between, especially around stomata as in E. schimperiana.;
smooth in anticlinal walls; (E) upright scale flakes in E. balsamifera

abaxial surface (hypostomatic leaves). The presence and

position of subsidiary cells are variable in Euphorbia
species. Subsidiary cells were found to be of unequal
size and shape whether the stoma was anisocytic,
paracytic, actinocytic or tetracytic. While some species
lack them around the stomata, others have more than
one arrangement of subsidiary cells.
The most common stomata type is anomocytic, while
the rare type is actinocytic, recorded only in E.
helioscopia. Moreover, sometimes stomata of more than
one type (up to three or four types) have been
encountered on the same leaf surface as in E. scordiifolia
(three types on the abaxial surface) and E. hirta (five

types on the abaxial and three on the adaxial). The

distribution and type of stomata are shown in Table 2.
In addition, in some species such as E. falcata and E.
helioscopia, the stomata are deeply sunken and
represented only by cuticular ledges surrounding the
minute pores as seen in ESEM.
Examples of species lacking subsidiary cells around
stomata are: E. peplus, E. chamaepeplus, E. granulata,
E. dracunculoides and E. acalyphoides. Therefore, they
all have anomocytic stomata.
On the other hand, E. balsamifera subsp. adenensis
has the mononcytic type on both leaf surfaces and
anisocytic and paracytic only on the abaxial surface. In

180 Int. Res. J. Plant Sci.

Table 2. Stomata type in Euphorbia taxa in Saudi Arabia.

Stomata
Subgenus

Anomocytic
Ad.*
Ab.*

Species
Tirucalli
Ermophyton
Esula

E. balsamifera subsp. adenensis

E. schimperi
E.sp. aff. consobrina
E. sp. aff. schimperi
E. cuneata

E. acalyphoides
E. pirottae

E. helioscopia
E. grossheimii
E. retusa
E. dracunculoides
E. falcate
E. peplus
E. chamaepeplus
E. schimperiana

Chamaesyce

E. Arabica
E. serpens
E. scordiifolia
E. granulate
E. inaequilatera
E. hirta
E. hypericifolia

*Ab. : abaxial surface

*Ad. : adaxial surface

Hemiparacytic
Ad.
Ab.

Paracytic
Ad.
Ab.

+
+

Anisocytic
Ad.
Ab.

+
+
+
+

+
+
+
+
+

Sunken
Deeply
Slightly
+
+
+
+
+

+
+

+
+

+
+

+
+

+
+
+

+
+
+
+
+

+
+
+
+

+
+
+

+
+
+

+
+
+
+
+

Actinocytic
Ad.
Ab.

+
+

Tetracytic
Ad.
Ab.

+
+

+
+

+
+
+
+

+
+
+
+
+
+

+
+

+
+
+

+
+

+
+

+
+

+
+

+
+
+

+ : stomata present

addition, E. schimperiana has anomocytic and

tetracytic on the abaxial only. In contrast, E.
cuneata has three types of stomata anomocytic,
paracytic and anisocytic only on the lower surface.
As well, E. aff. consobrina, E. aff. schimperi

and E. schimperi have paracytic and anisocytic

only in the abaxial on the first two and in both leaf
surfaces on the latter. Therefore, E. aff.
consobrina and E. aff. schimperi are the same
taxa. Then, comparing to E. schimperi results

from anatomy, we can also support that a new

subspecies of E. schimperi is identified.
Euphorbia serpens and E. scordiifolia are
similar
in
having
anomocytic,
paracytic
and anisocytic stomata, but they differ in their

Aldhebiani and Jury 181

Table.3. Venation type in Euphorbia taxa in Saudi Arabia.

Venation
Subgenus

Number of veins at
the leaf base

Species
1

Tirucalli
Ermophyton

Sheathed
Closed

Extended Tracheas

4 or more

Open

E.balsamifera
subsp.adenensis

E. schimperi

Rounded

E. aff. Schimperi

E. cuneata

E. acalyphoides

E. pirottae

Oval

Elongated

Esula

E. retusa

E. dracunculoides

E. falcate

E. peplus

E. chamaepeplus

E. schimperiana

Chamaesyce

E. Arabica

+
+

E. serpens

E. scordiifolia

E. granulate

E. inaequilatera

E. hirta

E. hypericifolia

E. cyathophora

Star shaped

E. grossheimii

E. helioscopia

Poinsettia

Margins

182 Int. Res. J. Plant Sci.

Figure 10. Sunken stomata on the abaxial surface:

1) Deeply sunken: (A) E. helioscopia, (B) E. falcata, (C) E. schimperiana.
2) Slightly sunken: (D) E. peplus, (E) E. pirottae.

Aldhebiani and Jury 183

Abaxial

Adaxial

A
Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

D
D

Figure 11. Stomata types in Euphorbia taxa in Saudi Arabia: (A) E. balsamifera subsp. adenensis, (B) E. schimperi,
(C) E. cuneata and (D) E. grossheimii.

184 Int. Res. J. Plant Sci.

Adaxial

Abaxial

AnomocyticAbax
ial
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

D
Figure 12. Stomata types in Euphorbia taxa: (A) E. falcata, (B) E. peplus, (C) E. chamaepeplus and (D) E.
schimperiana.

Aldhebiani and Jury 185

Adaxial

Abaxial

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

D
Figure 13. Figure 13. Stomata types in Euphorbia taxa in Saudi Arabia: (A) E. arabica, (B) E. serpens, (C)
E. scordiifolia and (D) E. helioscopia

distribution between the leaf surfaces (see Figure 13).

While E. hirta has the most types, E. helioscopia, E.
dracunculoides and E. acalyphoides have the least.
Regarding the position of the stomata on the surface
level of the epidermis, a variable degree of sunkeness
has been recorded. Using the ESEM, the stomata appear

deeply sunken in some species, whilst in others they are

only slightly sunken with the outer rims of the subsidiary
cells surrounding the aperture. The deeply and slightly
sunken species are recorded in Table 2. Additionally,
Figure 10 shows the sunken stomata in different species
under the ESEM.

186 Int. Res. J. Plant Sci.

Abaxial

Adaxial
Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Anomocytic
Hemiparacytic
Paracytic
Anisocytic
Tetracytic
Actinocytic

Figure 14. Stomata types in Euphorbia taxa in Saudi Arabia: (A) E. hirta, (B) E. hypericifolia, (C) E. pirottae and (D)
E.sp.aff. schimperi.

Venation
The venation patterns in the Euphorbia species under
investigation vary from one-veined, three-veined to those
with four or more veins. The predominant type is threeveined. However, in E. balsamifera subsp. Adenensis

there are seven middle parallel veins, three middle thick

and four thinner lateral ones (Figure 15 A and B).
Whereas in E. pirottae, there are three main middle
parallel veins and two thinner laterals (Figure 15 C). In
contrast, E. chamaepeplus, E. schimperiana and E.
arabica have only one thick main midrib coming from the

Aldhebiani and Jury 187

C
Figure 15. Figure 15: Venation patterns at base of leaves in
variants with more than three veins: (A & B) E. balsamifera
subsp. adenensis and (C) E. pirottae.

petiole then branched from the side into two lateral veins
at the point when the leaf starts widening (Figure 16).
Each of these veins branches again into secondary veins
at the leaf margin.

The rest of the species have just three veins: E.

grossheimii, E. dracunculoides, E. falcata, E. peplus, E.
serpens, E. scordiifolia, E. granulata, E. inaequilatera,
E. helioscopia, E. hirta, E. hypericifolia, and E.

188 Int. Res. J. Plant Sci.

Figure 16. Venation patterns with one main mid vein at the leaf base in (A) E. arabica and (B) E. chamaepeplus.

Figure 17. Venation patterns with three-veined leaves in (A) E. falcata and (B) E. granulata.

cyathophora. All the three veins come from the petiole

and start branching into secondary veins both toward the
margin and the mid vein direction (Figure 17 B).
The vein endings at the leaf margin vary from closed
to open. There is a relationship between the closed
margins and the sheathed veins (midrib and secondary
veins that are surrounded by parenchyma tissue). When
the veins are ornamented (sheathed), the margins are
closed. This can be seen in subgenus Chamaesyce, in
species such as: E. serpens, E. scordiifolia, E. granulata,
E. inaequilatera, E. hirta, E. hypericifolia and E. arabica.
Additionally, these species have the three-vein venation.
On the other hand, when the veins are not sheathed,
the veins ends are open at the leaf margin, as in: E.
balsamifera subsp. adenensis, E. schimperi, E. cuneata,
E. acalyphoides, E. grossheimii, E. dracunculoides, E.

falcata, E. peplus, E. chamaepeplus, E. schimperiana, E.

helioscopia, E. pirottae, E. aff. schimperi, and E.
cyathophora.
CONCLUSIONS
Anatomical characters are seen here to be very useful in
Euphorbia classification and identification. The results
obtained from anatomical studies were useful to group
some species and identify others.
Depending on the venation type, the Euphorbia
species under investigation are divided anatomically into
three main groups, see Figure 18.
Group one: veins are sheathed, epidermal cells
differentiated on adaxial and abaxial surfaces. Group two:

Aldhebiani and Jury 189

Figure 18. Hierarchial relation of Euphorbia taxa in Saudi Arabia based on anatomical characters.

190 Int. Res. J. Plant Sci.

veins are not sheathed and epidermal cells are alike in

both surfaces. And the third group includes species that
do not belong to any of the previous groups. The first
group can be further divided into three-veined and oneveined, while the second group into three-veined and four
or more veins.
In addition, E. grossheimii, E. retusa and E. pirottae
are related by being three-veined, open at the margins,
with rounded to oval tracheas and elongated cells both
on adaxial and abaxial surfaces. However, in E. retusa
the epidermal cells have straight cell walls on the adaxial
surface. They are also similar in having hemiparacytic
and paracytic types of stomata either on the adaxial or
the abaxial surface.
On the other hand, species with two different
epidermal cells on the leaf surfaces usually have threesheathed veins with closed margins and elongated
trachea, such as: E. serpens, E. scordiifolia, E. granulata,
E. inaequilatera, E. hirta and E. hypericifolia which belong
to subgenus Chamaesyce according to (Carter and
Smith, 1988).
In the case of E. granulata, which has been
described as having three different variants: E. granulata
var. granulata (both leaf surfaces hairy), E. granulata var.
glabrata (adaxial glabrous and abaxial hairy) and a
glabrous variant (the whole plant glabrous). Anatomical
work has shown, especially by using the ESEM, that the
leaves are either hairy or glabrous on both surfaces. So,
when the leaf is hairy, both surfaces have differences in
hair distribution. Usually, the lower surface is more hairy
than upper surface (E. granulata var. glabrata), but
sometimes they are evenly distributed (E.granulata var.
granulata). In addition, hairs on both surfaces are
multicellular. In contrast, when the leaf is glabrous, both
leaf surfaces are glabrous and the whole plant is
glabrous (suggested new variety). They were similar in
having the same venation type (three-veined).
Finally, E. aff. schimperi has some similarity with E.
schimperi in having the same open margin not the
ornamented venation type. As well, they both have the
same type of stomata paracytic and anisocytic but they
differ in their distribution on abaxial and adaxial. Thus,
this may support the suggestion considering E. aff.
schimperi as a new subspecies of E. schimperi.
REFERENCES
Baranova M (1992). Principels of comparative stomatographic studies
of flowering plants. The Botanical Review 58: 49-99.
Carlquist S (1961). Comparative Plant Anatomy. Holt, Rinehart &
Winston, New York.
Carter S, Smith AR (1988). Euphorbiaceae (Part2). In: Polhill, ed. Flora
of Tropical East Africa. Rotterdam, Brookfield: A.A. Balkema. 409597.
Chaudhary SA (2001). Flora of The Kingdom of Saudi Arabia Vol. 3.
Ministry of Agriculture and Water Riyadh.
Dehay C (1935). Lppareil Libero-ligneux foliare des Euphorbiaceae.
Annales des Sciences Naturelles Botanique 10: 147-190.

Dehgan B (1982). Comparative anatomy of the petiole and infrageneric

relationships in Jatropha (Euphorbiaceae). American Journal of
Botany 69: 1283-1295.
Gaucher L (1898). tude anatomique du genre Euphorbia L.P.
Klincksieck, Paris.
Gaucher L (1902). Recherches anatomiques sur les Euphorbiaces.
Annales des Sciences Naturelles Botanique 15: 161-309.
Gaucher L (1902). Recherches anatomiques sur les Euphorbiaces.
Annales des Sciences Naturelles Botanique 15: 161-309.
Govaerts R, Fordin D, Radcliffe-Smith A (2000). World Checklist and
Bibliography of Euphorbiaceae. The Royal Botanic Gardens, Kew.
Hickey LJ (1973). Classification of the architecture of dicotyledonous
leaves. American Journal of Botany 60: 17-33.
Hickey LJ (1979). A revised classification of the architecture of
dicotyledonous leaves. In: Metcalfe CR and Chalk L, eds. Anatomy of
Dicotyledons, Systematic Anatomy of the leaf and stem. 2nd ed.
Clarendon Press, Oxford:. 25-39.
Kakkar L, Paliwal GS (1972). Epidermis in Euphorbia. Indian Science
Congress Association Proceedings 59: 326-327.
Kakkar L, Paliwal GS (1974). Studies on the Leaf Anatomy of Euphorbia
Part 5 Epidermis. Proceedings of the Indian National Science
Academy Part B Biological Sciences 40: 55-67.
Khler E (1993). Blattvervatur-Muster der Buxaceae Dumortierr und
Simmondsiaceae Van Tieghem. Feddes Repertorium 104: 145-167.
Levin GA (1986a). Systematic foliar morphology of Phylanthoideae
(Euphorbiaceae). II. Phenetic analysis. Annals of Missouri Botanical
Garden 73: 86-98.
Levin GA (1986b). Systematic foliar morphology of Phylanthoideae. III
Cladistic analysis. Systematic Botany 11: 515-530.
Levin GA (1986c). Systematic foliar morphology of Phyllanthoideae
(Euphorbiaceae). I conspectus. Annals of Missouri Botanical Garden
73: 29-85.
Mahlberg P (1973). Histochemistry and Scanning Electron Microscopy
of Starch Grains from Latex of Euphorbia-Terracina and EuphorbiaTirucalli. Proceedings of the Indiana Academy of Science 82: 132.
Mahlberg PG (1974). Scanning Microscopic Comparison of Starch
Grains from the Latex of Euphorbia-Spp. Proceedings of the Indiana
Academy of Science 83: 83-84.
Mahlberg PG (1975). Evolution of the Laticifer in Euphorbia as
Interpreted from Starch Grain Morphology. American Journal of
Botany 62: 577-583.
Mahlberg PG (1982). Comparative Morphology of Starch Grains in
Latex from Varieties of Poinsettia Euphorbia-Pulcherrima
Euphorbiaceae. Botanical Gazette 143: 206-209.
Mahlberg PG, Davis DG, Galitz DS, Manners GD (1987). Laticifers and
the Classification of Euphorbia the Chemotaxonomy of EuphorbiaEsula L. Botanical Journal of the Linnean Society 94: 165-180.
Metcalfe C, Chalk L (1950). Anatomy of the Dicotyledons. At the
Clarendon Press, Oxford.
Miller K, Webster GL (1962). Systematic position of Cnidoscolus and
Jatropha. Brittonia 14: 174-180.
Pax F (1884). Die Anatomie der Euphorbiaceen in ihrer Beziehung zum
System derselben. Botanische Jahrbcher fr Systematik,
Pflanzengeschichte und Pflanzengeographie. Leipzig 5: 384-421.
Payne WW (1970). Helicocytic and allelocytic stomata: unrecognized
patters in the Dicotyledonae. Am. J. Bot.57: 140-147.
Pray TR (1955a). Foliar venation of angiosperm. II. Histogenesis of the
venation of Liriodendron. Am. J. Bot. 42: 18-27.
Pray TR 1955b. Foliar venation of angiosperms. III. Pattern and
histology of the venation of Hosta. Am. J. Bot. 42: 11-18.
Radford AE, Dickison WC, Massey JR, Bell CR (1974). Vascular Plant
Systematics. Harper & Row Publishers, London, New York.
Radlkofer L (1870). Ueber Pausandra, ein neues EuphorbiaceenGenus. Flora 53: 81-95.
Raju VS, Rao PN (1977). Variation in the structure and development of
foliar stomata in the Euphorbiaceae. Botanical Journal of the Linnean
Society 75: 69-97.
Raju VS, Rao PN (1987). The Taxonomic Use of the Basic Stomatal
Type in the Generic Delimitation of Chamaesyce Euphorbiaceae.
Feddes Repertorium 98: 137-142.

Aldhebiani and Jury 191

Rosowski JR (1968). Laticifer Morphology in the Mature Stem and Leaf

of Euphorbia supina. Botanical Gazette 129: 113-120.
Roth-Nebelsick A, Uhl D, Mosbrugger V, Kerp H (2001). Evolution and
function of leaf venation architecture: a review. Annals of Botany 87:
553-566.
Sehgal L, Paliwal GS (1974). Studies on the Leaf Anatomy of Euphorbia
Part 2 Venation Patterns. Botanical Journal of the Linnean Society
68: 173-208.
Singh G (2004). Plant Systematics: An Integrated Approach. Science
Publishers, Enfield.
Solereder H (1899). Systematische Anatomie der Dicotyledonen.
Ergnzungsband, Enke, Stuttgart.
Solereder H (1908). Systematische Anatomie der Dicotyledonen.
Ergnzungsband, Enke, Stuttgart.

Stace CA (1965). Cuticular studies as an aid to plant taxonomy. Bulletin

of the British Museum Natural History Botany 4: 3-78.
Stace CA (1989). Plant Taxonomy and Biosystematics. Hodder &
Stoughton, London.
Tognini F (1897). Contribuzione allo studio della organogenia
comparata degli stomi. Institut de Botanique, University de Geneve 4:
1-42.
Van Cotthem WR (1970). A classification of stomatal types Botanical
Journal of Linnean Society 63: 235-246.
Wagner W (1979). Reticulate veins in the systematics of modern ferns.
Taxon 1: 87-95.
Webster GL (1994a). Classification of the Euphorbiaceae. Annals of the
Missouri Botanical Garden 81: 3-32.