Int. J. LifeSc. Bt & Pharm. Res. 2012
Amal Kumar Mondal and Sayantan Tripathi, 2012
ISSN 2250-3137 www.ijlbpr.com
Vol. 1, No. 3, July 2012
© 2012 IJLBPR. All Rights Reserved
Research Paper
COMPARATIVE (QUANTITATIVE AND
QUALITATIVE) STUDIES OF STOMATA
OF SELECTED SIX MEDICINALLY
VIABLE SPECIES OF CASSIA L.
Sayantan Tripathi1 and Amal Kumar Mondal1*
*Corresponding Author: Amal Kumar Mondal,  [email protected]
The Stomatal diversity (size, shapes, types and orientation) in the foliar epidermis has great
value in plant systematics studies. The present paper deals with comparative study of stomatal
structure of six species of Cassia which are used by rural people of South West Bengal as
medicine. Leaf epidermal studies mainly stomatal studies of the 6 species of Cassia of family
Caesalpiniaceae viz., Cassia alata L., Cassia fistula L., Cassia occidentals L., Cassia siamea
Lamk., Cassia sophera L., Cassia tora L. were made. Leaf clearings and cuticular preparations
were examined with light microscopy. The study is based on the presence and absence of
stomata, types of stomata present in the epidermal surface, stomatal count/cm2, stomatal index
and epidermal cell shape. The size and shapes of stomata are also varied in the tree species
which bear larger size of stomata in respect of other habits and habitats. Three types of stomata
were observed viz. paracytic, anisocytic and anomocytic. Among these 3 types of stomata the
paracytic type of stomata are more common and than other.
Keywords: Epidermis, Medicinal use, Cassia, Paracytic stomata
INTRODUCTION
lives in rural areas that mostly depend on Unani
system of medicines (Soomro et al., 1997). The
Various plants and its parts are used in medicinal
purpose from ancient age. Now a days it is
available literature shows that leaf epidermal
observed that the African gorillas also use various
features are important in systematic botany
plants and its parts to cure their health. The
similar to the use of modern techniques and
medicinal plants have been used by Hakims and
chemical composition (Edeoga and Ikem, 2001;
in folklore medicines as 80% of the population
and Mbagwu and Edeoga, 2006). Epidermal
1
Plant Taxonomy, Biosystematics and Molecular Taxonomy Laboratory, Department of Botany & Forestry, Vidyasagar University.
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Int. J. LifeSc. Bt & Pharm. Res. 2012
Amal Kumar Mondal and Sayantan Tripathi, 2012
structures and stomatal ontogeny of some
et al., 2007; Kaplan et al., 2007; Keshavarzi and
Nigerian ferns have been found relevant in their
recognition (Gill and Karatela, 1985).
Zare, 2006). Despite the immense economic
Many workers such as Edeoga (1991),
Edeoga and Osawe (1996), Mbagwu and Edeoga
(2006), Nwachukwu and Mbagwu (2006)
stressed that epidermal and cuticular traits of
plants epidermal cells, type and arrangement of
stomata, size and shape of trichomes and
number of vascular bundles could serve as vital
tools in solving taxonomic problems in
Angiosperms.
importance of the stomatal apertures, reports on
importance of the legumes and the physiological
the frequency and the structure of the stomata
are lacking or incomplete for many species.
On the basis of arrangement of epidermal all
neighbouring the guard all, more than 25 main
types of stomata in dicots have been recognized
(Metcalfe and Chalk, 1979). Stace (1980) reported
31 different types of stomata among cotyledonous
plants. But the present study is based on the
Stomatogenesis has long been studied by
morphologists, physiologists and taxonomist .The
morphology and ontogenies of taxa are important
in intrageneric systematics. Diversity in stomata
types, even on the same surface of an organ,
indicates the weakness in using stomata as a
taxonomic character. (Pant and Kidwai, 1964). In
spite of diversity, the most frequent stomata type
can be used as a taxonomic character (Gopal,
1970).
paper of Metcalfe and Chalk (1950) who described
the 4 types – i.e. Anisocytic, Anomocytic, Diacytic
and Paracytic.
The aim of present study is to use stomatal
characters as aid in taxonomy of these selected
medicinally useful Cassia species. The study
would help in the identification and authentication
of these medicinal plants on the basis of
stomatogenesis.
Caesalpiniaceae is a family of dicotyledonous
MATERIAL AND METHODS
plant with about 160 genera and some 2,000
species. The genus Cassia is one of the genuses
of Caesalpiniaceae having about 692 species.
Some of the Cassia species is used as medicine
Plant Material: The medicinal plant specimens
Cassia alata L., Cassia fistula L., Cassia
occidentals L., Cassia siamea Lamk., Cassia
sophera L. and Cassia tora L. belonging the family
Caesalpiniaccae under order Leguminales were
taken for stomatal study.
in Eastern India mainly South West Bengal. We
select Cassia alata L., Cassia fistula L., Cassia
occidentals L., Cassia siamea Lamk., Cassia
sophera L., Cassia tora L. which are frequently
Cassia alata L. is popular in traditional
treatment of asthma, leprosy and it is used for
controlling of ringworm (Paria, 2005).
used by the rural and tribal people of south West
Bengal.
Apart from physiognomic characters,
Role of Cassia fistula L. is used as purgative
and also used to treat skin disease. Its leaves
are used to treat skin disease and ulcer. Powder
of Cassia fistula L. root is given to cure menstrual
anatomical properties of plant parts are sources
for taxonomic inferences in different groups of
flowering plants (Edeoga et al., 2007; Guimeraes
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Int. J. LifeSc. Bt & Pharm. Res. 2012
Amal Kumar Mondal and Sayantan Tripathi, 2012
Isolation of Epidermis: Epidermis of leaf is
isolated from both fresh and dry plant specimen.
The mature leaves were fixed in FAA solution
(acetic acid: alcohol: formalin: water = 2:5:1:12)
for 24 hours and washed in 70% ethanol. Three
circular disk samples were cut from an area
adjacent to the midrib of each leaf. Disk sample
was boiled in 5% aqueous solution of KOH for
5-10 minute. Epidermal peals were stripped and
stained with 1% in 50% aqueous ethanol,
saffranin and temporary mount in glycerine.
disorder and its pulp of seed is used in high blood
pressure (Paria, 2005).
Cassia ossidentales L. is popularly used as
purgative. It is also used to treat hysteria,
whooping cough and urinary troubles (Paria,
2005).
Cassia sophera L. is an annual erect shrub of
important medicinal properties. The whole plant
extracts and leaves have expectorant properties,
cures cough, asthma and acute bronchitis. They
are specific to eliminate ring worms and also
useful in the treatment of gonorrhoea and syphilis.
The bark is used in the treatment of diabetes; the
roots in elephantiasis, wounds. (Kirtikar and
Basu,1935).
Stomatal frequency counts made and camera
lucida were drawn. The numbers of stomata were
counted in each field (0.001386 cm 2). The
stomatal frequency was based on average
obtained from observations of 3 microscopic
fields. Stomatal index (I) was calculated by the
following formula using the no. of stomata(s)
and epidermal cells was present in a unit area
[I=S/(S+E)].
The leaves of Cassia siamea Lamk. are used
in the treatment of diabetes, disturbances in the
bodily functions, lymph node swelling, urine
stones, general deficiency conditions, beri beri ,
classic deficiency of avitaminose by lack of
vitamin B1 (thiamine) in gastrointestinal disorders
- malabsorption - meals taken with polished rice
etc., antihypertensive, insomnia (sleeplessness),
against dysentery and disorders of the large
intestine. (Kirtikar and Basu, 1935).
After preparing, slides were observed under
light microscope [(40x)Leica DM 1000] and phase
contrast microscope for detailed analysis and
obtaining better picture as well as measuring the
length and breadth of stomata including guard
cells.
Cassia tora L. is popularly known for the
treatment of dysentery and eye disease. Leaves
are also used in boils, ring worm, leprosy, bronchitis
and liver complaints. Pest of fresh pods is applied
to cure lentigo. Seed powder is taken to cure
intestinal problems caused by intestinal worms.
Seed pest mixed with boiled rice water is applied
in case of one sided headache (Paria, 2005).
RESULTS AND DISCUSSION
The results in this investigation were summarised
in tables including stomata count, stomata type,
stomatal index, stomatal complex.
The entire selected species share a number
of common characters, i.e., shape of epidermal
cell, presence of stomata, stomatal index and
stomatal complex which are useful in
The species identification of the selected
material was determined according to standard
literature. It was done in the months from February
to April. The foliar epidermal peals were taken
from the middle of both surfaces of mature leaves.
distinguishing these species.
Epidermal Cell: The shape of foliar epidermis is
one of the significant taxonomic characters.
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Int. J. LifeSc. Bt & Pharm. Res. 2012
Amal Kumar Mondal and Sayantan Tripathi, 2012
abundant stomata were present on abaxial
surface as compared to adaxial surface.
Taxonomic studies of a number of families are
based on leaf epidermis anatomy (Bhatia, 1984,
Jones, 1986). In this investigation it was found that
the epidermal cell shape varies slightly with growth
habit. The shrub species posses pentagonal to
polygonal shaped epidermal cells. The epidermal
cell shapes of tree species are variously shaped
or undulating to irregular (Table 1).
All these selected species bear paracytic
stomata except the adaxial surface of Cassia
alata L. (Figure 2A). The adaxial surface of Cassia
alata L. (Figure 2A) bears anisocytic stomata.
Anisocytic and paracytic stomata were also found
in the abaxial surface of Cassia occidentals L.
(Figure 1B), Cassia sophera L. (Figure 1F) and
Cassia tora L. (Figure 1D). From this investigation
it was found that all the anisocytic stomata
bearing species are shrub in growth habit.
Anomocytic stomata are found only in the abaxial
surface of Cassia siamea Lamk. (Figure 2F). All
the investigated species showed paracytic
stomata, a mixture of stomata in same species
were also observed. Cassia ocidentales L., Cassia
sophera L. and Cassia tora L. are closely related.
They showed paracytic stomata in adaxial surface
and in the abaxial surface they showed paracytic
stomata along with anisocytic type (Table 1).
In these six species only Cassia siamea Lamk.
(Figure 2E, F) bears hypostomatic leaves i.e.
stomata present only at the abaxial surface of
leaves. Here it was found that the shape of
epidermal cells varies in their surfaces. The
adaxial surface of the leaf shows tetragonal to
polygonal shaped epidermal cells but the abaxial
surface i.e. stomata bearing surface shows
undulating to irregular shaped epidermal cells.
Stomata: Three different types of stomata
anomocytic, anisocytic, paracytic were observed
in these selected species. It was found that
Table 1: Qualitative Foliar Stomatal Characteristics of the Selected Species of Cassia
S. No.
1.
2.
3.
4.
5.
6.
Taxa
Cassia alata L.
Cassia fistula L.
Cassia occidentals L.
Cassia siamea Lamk.
Cassia sophera L.
Cassia tora L.
Stomata
Stomata Type
Anomo- Aniso- Paracytic
cytic
cytic
Habit
Surface
Shrub
Ad.
Pentagonal to hexagonal
+
–
+
–
–
Ab.
Pentagonal to hexagonal
+
–
+
+
–
Ad.
Variously shaped
+
–
–
+
–
Ab.
Variously shaped
+
–
–
+
–
Ad.
Pentagonal to polygonal
+
–
–
+
–
Ab.
Pentagonal to polygonal
+
–
+
+
–
Ad.
Tetragonal to polygonal
–
–
–
–
–
Ab.
Undulating to irregular
+
+
–
+
–
Ad
Pentagonal to polygonal
+
–
–
+
–
Ab
Undulating to irregular
+
–
+
+
Ad.
Pentagonal to polygonal
+
–
–
+
–
Ab.
Pentagonal to polygonal
+
–
+
+
–
Tree
Shrub
Tree
Shrub
Shrub
Leaf Epidermal Cell Shape
Presence
of
Stomata
107
Diacytic
Int. J. LifeSc. Bt & Pharm. Res. 2012
Amal Kumar Mondal and Sayantan Tripathi, 2012
Table 2: Quantitative Foliar Stomatal Characteristics
of the Selected Species of Cassia
S. No.
1.
2.
3.
4.
5.
6.
Taxa
Cassia alata L.
Cassia fistula L.
Cassia occidentals L
Cassia siamea Lamk.
Cassia sophera L.
Cassia tora L.
Habit
Surface
Stomatal Complex. Legth × Width (with guard cell) µm2
Shrub
Ad
23.89(21.28-25.77)×20.27(18.43-22.41)
Ab
18.48(17.46-19.31)×12.36(10.47-14.12)
Ad
13.74(13.04-14.65)×11.66(11.32-12.59)
Ab
15.22(12.5-17.5)×10.25(7.75-12.5)
Ad
23.11(22.31-23.81)×16.49(15.67-17.11)
Ab
22.98(21.65-25.48)×17.09(14.28-20.99)
Ad
–
Ab
23.49(21.34-25.40)×15.32(14.49-16.71)
Ad
24.85(23.61-26.47)×15.24(14.47-16.10)
Ab
22.54(21.24-24.55)×15.29(14.54-16.35)
Ad
22.10(19.99-23.16)×13.06(11.84-13.92)
Ab
23.28(23.27-23.29)×15.66(15.17-16.38)
Tree
Shrub
Tree
Shrub
Shrub
Table 3: Stomatal Index Percentage and Stomatal Count/cm2
of the Selected Species of Cassia
S. No.
1.
2.
3.
4.
5.
6
Taxa
Cassia alata L.
Cassia fistula L.
Cassia occidentals L
Cassia siamea Lamk.
Cassia sophera L.
Cassia tora L.
Habit
Surface of Leaflet
Stomatal Index (SI) Percentage
Stomata Count/cm2
Shrub
Ad
8.3
6475
Ab
12.5
7914
Ad
13.3
8734
Ab
16.8
11511
Ad
21.7
12950
Ab
28.7
14388
Ad
–
–
Ab
22.6
13670
Ad
18
11511
Ab
23.1
19424
Ad
23.3
10072
Ab
24.3
12950
Tree
Shrub
Tree
Shrub
Shrub
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Amal Kumar Mondal and Sayantan Tripathi, 2012
Figure 1: Leaf Epidermis Showing Stomata of
A. Cassia occidentalis (adaxial surface); B. Cassia occidentalis (abaxial surface);
C. Cassia tora (adaxial surface); D. Cassia tora (abaxial surface);
E. Cassia sophera (adaxial surface); and F. Cassia sophera (abaxial surface)
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Amal Kumar Mondal and Sayantan Tripathi, 2012
Figure 2: Leaf Epidermis Showing Stomata of
A. Cassia alata (adaxial surface); B. Cassia alata (abaxial surface);
C. Cassia fistula (adaxial surface); D. Cassia fistula (abaxial surface);
E. Cassia siamea (adaxial surface); and F. Cassia siamea (abaxial surface)
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Amal Kumar Mondal and Sayantan Tripathi, 2012
stomatal characters of 6 species of Cassia
Quantitative analysis of stomatal complex in
the selected species shows great variation (Table
2). Here it was found that except Cassia fistula L.
Cassia alata L., Cassia occidentals L., Cassia
siamea Lamk., Cassia sophera L., Cassia tora
L. share almost equal size of stomata which
shows close relationship. In these 5 species
Cassia sophera L. and Cassia occidentals L.
share almost equal size of stomata and the ratio
between size of stomata of adaxial and abaxial
surface is almost equal. Smallest stomata were
found in the adaxial surface of Cassia fistula L.
(13.74×11.68 µm2).
belonging to family Caesalpniaceae under the
order Leguminales, used in medicinal purpose in
the South West Bengal region.
From the study it can be concluded that there
is no relationship between the stomata size and
growth habit. Size of stomata and epidermal cell
shape prove that all the investigated species are
closely related. All the investigated species
showed paracytic stomata, a mixture of stomata
in same species were also observed. Cassia
ocidentales L., Cassia sophera L. and Cassia tora
L. are closely related. They showed paracytic
stomata in adaxial surface. In the abaxial surface
Stomatal index is one of the useful tools in order
to distinguish species. It was found that stomatal
index has low value on adaxial surface as
compared to abaxial surface (Table 3). According
to stomatal index Casia alata L. and Cassia fistula
L. are closely related. Cassia occidentals L.,
Cassia siamea Lamk., Cassia sophera L. and
Cassia tora L. showed relationship among them.
Highest value of stomatal index was observed
on abaxial surface of Cassia occidentals L. (28.7),
where lowest value was observed on the adaxial
surface of Cassia alata L. (8.3).
they showed both anisocytic and paracytic type
of stomata. Shape of epidermal cells, presence
of stomata, stomatal type, stomatal complex,
stomatal index were investigated. These
parameters are helpful to differentiate the species.
However, the stomatal features may prove to be
a little taxonomic value unless the developments
of different stomata types were studied. A greater
number of information on taxa will be helpful to
understand the taxonomic value of stomata type
and distribution.
Stomatal count is a significant character to
differentiate various species. Highest stomata
count per unit area was found on the abaxial
surface of Cassia sophera L. (194245/cm 2).
Lowest stomata count was found in adaxial
surface of Cassia alata L. (6475/cm2). All the
investigated Cassia species share more or less
equal number of stomata which shows they are
closely related.
ACKNOWLEDGMENT
We are grateful to our honourable Vice Chancellor
of Vidyasagar University, Prof. Ranjan
Chakraborti, and we are also thankful to our
friends (Santanu Dash and Dulal Chandra Bera).
Without their help we cannot complete our work.
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