Not. Bot. Hort. Agrobot.

Cluj, 2007 Volume 35, Issue 2

Print ISSN 0255-965X; Electronic ISSN 1842-4309

COMPARATIVE FOLIAR EPIDERMAL STUDIES IN CYMBOPOGON

CITRATUS (STAPF.) AND CYMBOPOGON GIGANTEUS
(HOCHST.) CHIOV. IN NIGERIA
Folorunso A. E., O. A. Oyetunji
Department of Botany, Obafemi Awolowo University, Ile-Ife,
Nigeria; e-mail: afolorun2002@yahoo.co.uk
Abstract. The foliar epidermal studies were carried out on Cymbopogon citratus and
Cymbopogon giganteus with the aim of determining the patterns of variation in their epidermal
characteristics and assessing their value in species identification and classification. The characters of
diagnostic importance in the identification of C. citratus are the micro hairs, which are sparsely
distributed in the adaxial epidermis and prickle hairs present in both abaxial and adaxial epidermis.
The diagnostic character for C. giganteus is the papillae seen alongside their long cells.
Key words: epidermal, Cymbopogon, micro hair, prickle hair, papillae
INTRODUCTION

The genus Cymbopogon belongs to the grass family, Poaceae which is a very large
cosmopolitan family consisting of about 50 to 60 tribes, 660 genera and 9000 species
throughout the world (Olorode, 1984; Hutchinson and Dalziel, 1972). Cymbopogon is a genus
of about 55 species of grasses, native to warm temperate and tropical regions of the world. It
consists of tall grasses. It is composed of tufted perennial grasses often with aromatic leaves.
The inflorescences have two branches with the common stalk not longer than the branches
and usually hidden within the spathe. Usually, the inflorescences are very densely clustered
in large panicles. Two species grow wild in Nigeria, and a third, commonly known as
Cymbopogon citratus (lemon grass) is often cultivated as a border to roads and paths.
The economic importance of Cymbopogon citratus includes its use in aromatherapy
due to the strong and sweet aroma present in it. Cymbopogon citratus (lemon grass) both
deodorizes and is an effective antiseptic. It is excellent for tired and aching feet. Lemon grass
oil may be used in the treatment of acne, athletes foot, excessive perspiration, flatulence,
insect- repellant, muscle aches, oily skin, scabies and stress. Safety precautions should be
taken when using Cymbopogon citratus in aromatherapy and it should not be used on
children. Those with glaucoma should avoid use and caution should be used in prostatic
hyperplasia and with skin hypersensitivity or damaged skin.
Cymbopogon citratus (lemon grass) is widely used as an herb in Asia. It has a lemony
flavour and can be dried and powdered or used fresh. The stalk itself is too hard to be eaten,
except for the soft inner part. However, it can be finely sliced and added to recipes. It may
also be bruised and wholly added as this releases the aromatic oils from the juice sacs in the
stalk. It is commonly used in teas, soups and curries. It is also suitable for poultry, fish and
seafood. In India, Cymbopogon citratus is used both as a medical herb and in perfumes.
Artemisia absinthium and Cymbopogon citratus possess strong allelopathic inhibitory effects
on Bidens pilosa (Cruz, 2002).

The hydro-distilled essential oil from the leaves of Cymbopogon citratus grown in
Zambia was analysed by GC and GC- MS. Sixteen compounds representing 93.4 % of the oil
were identified, of which geranial (39.0 %), neral (29.4 %) and myrcene (18.0 %) were the
major components. Small amounts of geraniol (1.7%) and linalol (1.3%) were also detected
(Chisowa et al., 1998). Cymbopogon citratus (lemon grass) fertilized with 40 KgN and 40 Kg
P2O5/ha has led to high herb yield (256.0 g/ha). There was no response to potassium
fertilization (Prasad and Mukherji, 1980).
The grass family Poaceae, is noted for its wide diversity and complexity and so has
posed many problems to the taxonomists using the traditional methods based on gross
morphology (Srivastava, 1978). Before the later part of the 19th century, taxonomists were
confined to the use of the features of reproductive organs, as floral characters were considered
to provide the most valuable characters to taxonomic affinities (Nwokeocha, 1996). Of all the
non-reproductive organs, the leaf is the most widely used in plant taxonomy (Stace, 1965,
1984). Srivastava (1978) described the leaf epidermis as the second most important character
after cytology for solving taxonomic problem.
Following the works of Metcalfe and Chalk (1950) and Metcalfe (1954) which today
serve as standard references to plant anatomy, the use of vegetative anatomical characters in
taxonomy became a routine procedure. Since then, the immense value of the leaf epidermis
and vegetative anatomy in grass systematics (part of which is the genus, Cymbopogon) has
been demonstrated by many workers among whom are Tomlinson (1959); Srivastava (1978);
Renvoize (1982, 1987); Ogundipe and Olatunji (1989, 1991).
The aim of this study is to determine the patterns of variation in epidermal
characteristics, assess their value in species identification and classification and also using the
epidermal studies in establishing the taxonomic relationships between the two species of
Cymbopogon.
MATERIALS AND METHODS

The epidermal preparations of the adaxial and abaxial surfaces of the leaf blade were
made in ten accessions each of Cymbopogon citratus collected from the Agricultural farm of
Obafemi Awolowo University, Ile-Ife, Osun State. Herbarium specimens were used in the
case of Cymbopogon giganteus. The leaves of those herbarium specimens were boiled in
water for 5-10 minutes to soften them. The scraping method (Cutler, 1978) was used for
Cymbopogon citratus and Cymbopogon giganteus. The specimens were decolourised in about
5% solution of Sodium hypochlorite (domestic bleach) for 30-60 minutes. The cleared
epidermal peels were preserved in 50% ethanol, stained in Alcian blue and counter-stained in
Toluidine blue, to enhance contrast. To further enhance the identification of cork and silica
cells, some peels were stained in 1% Sudan IV solution. All the preparations were mounted in
25% glycerol. Both adaxial and abaxial surfaces of the leaves were studied.
Illustrations of the foliar epidermal features were done by camera lucida under 25
objective power of leitz DIALUX research microscope. Anatomical measurements were made
in ocular units at 40 objective power and converted to micrometers.
The stomatal Index (I) was estimated for the two leaf surfaces using the formula:
S
Stomatal Index (I) =
x 100
E + S
SI = Stomatal index
S = number of stomata per unit area
E = number of ordinary epidermal cells plus subsidiary cells in the same unit area
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RESULTS AND DISCUSSIONS

Cymbopogon citratus (abaxial) (Figure 1a and Table 1)

Long cells: rectangular, conspicuously elongated (many times longer than broad); 5-7 rowed
between the veins; cell wall wavy; papillae not seen.
Short cells: present over and between the veins; solitary and sometimes paired; arranged in
rows of 2-3 cells.
Prickle hair: present mostly with pointed tip and rather elongated, swollen bases; very
frequent to numerous and arranged in rows.
Unequal angular prickles also present on leaf margins.
Micro hair: none seen.
Macro hair: none seen.
Stomata: large, infrequent stomata; dome shaped stoma; 2-banded between the veins;
paracytic stomata type seen.
Anticlinal wall: straight
Silica bodies: seen and cross shaped.
Table 1
Some descriptive statistics of leaf epidermal attributes of Cymbopogon citratus
Trait
(m)
Length of stoma
Width of stoma
Length of long cells
Width of long cells
Length of short cells
Width of short cells
Length of prickle hairs
Width of prickle hairs
a = Adaxial
b = Abaxial

Minimum
a
b
2.88
3.24
2.16
2.88
5.04
9.72
1.80
2.16
0.72
0.72
1.44
2.16
2.52
3.24
0.72
0.72

Maximum
a
b
3.24
3.60
2.52
3.60
6.84
14.40
2.52
2.52
1.08
1.08
1.80
2.88
3.60
5.04
1.08
1.08

Mean
a
2.95
2.30
5.83
2.02
0.86
1.58
3.02
0.86

b
3.31
3.24
11.66
2.38
0.79
2.66
3.96
0.86

Standard Dev.
a
b
0.14
0.14
0.18
0.23
0.62
1.74
0.29
0.18
0.18
0.14
0.18
0.29
0.37
0.68
0.18
0.18

Standard Error
a
b
0.06 0.06
0.08 0.10
0.28 0.78
0.13 0.08
0.08 0.06
0.08 0.13
0.17 0.30
0.08 0.08

Figure 1 a. The leaf epidermal of Cymbopogon citratus. Abaxial epidermis; ap angular prickle;
ph prickle hair; sc short cell; lc long cell; sb silica body; s stomata

Cymbopogon citratus (adaxial) (Figure 1b and Table 1)

Long cells: rectangular, slightly elongated (longer than broad); 8-10 rowed between the veins;
the long cells have thin, wavy walls.
Short cells: mostly solitary, sometimes paired and arranged in rows of 4-6 cells.
Prickle hair: seen; frequent to numerous with pointed tip and elongated swollen bases.
Micro hair: seen; about 18.36-18.72m long and 1.08m broad.
Macro hair: none seen.
Stomata: large, frequent, dome shaped stoma; 2-4 banded between the veins; paracytic
stomata type seen.
Anticlinal wall: wavy
Silica bodies: seen; mostly cross shaped

Figure 1 b. The leaf epidermal of Cymbopogon citratus. Adaxial epidermis; ph prickle hair;
sc short cell; lc long cell; sb silica body; s stomata; mh micro hair

Cymbopogon giganteus (abaxial) (Figure 2a and Table 2)

Long cells: rectangular, conspicuously elongated (long than broad); 4-7 rowed between the
veins; cell wall of the long cells are thin and sinous.
Short cells: present over the veins in rows of 4-6 or more cells.
Prickle hair: none seen.
Micro hair: none seen.
Macro hair: none seen.
Stomata: large, infrequent stomata; 2-banded between the veins; dome shaped stoma;
paracytic stomata type seen.
Anticlinal wall; straight.
Silica bodies; seen and mostly cross shaped.

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Table 2
Some descriptive statistics of leaf epidermal attributes of Cymbopogon giganteus
Trait
(m)
Length of stoma
Width of stoma
Length of long cells
Width of long cells
Length of short cells
Width of short cells
a = Adaxial
b = Abaxial

Minimum
a
b
3.24
2.52
2.16
1.08
3.60
6.12
1.80
2.52
0.72
0.72
1.80
2.16

Maximum
a
b
3.60
4.32
2.52
2.88
6.48
10.80
2.52
6.84
1.44
1.08
2.16
2.88

Mean
a
3.31
2.30
4.90
2.02
1.01
1.73

b
3.53
1.94
7.85
3.96
0.86
2.59

Standard Dev.
a
b
0.24
0.62
0.18
0.67
0.98
1.70
0.29
1.53
0.27
0.18
0.42
0.27

Standard Error
a
b
0.11 0.28
0.08 0.30
0.44 0.76
0.13 0.68
0.12 0.08
0.20 0.12

Figure 2 a. The leaf epidermal of Cymbopogon giganteus. Abaxial epidermis;

sc short cell; lc long cell; sb silica body

Cymbopogon giganteus (adaxial) (Figure 2b and Table 2)

Long cells: rectangular, slightly elongated (almost twice longer than broad with the width
more or less uniform); 9-11 rowed between the veins; cell wall predominantly sinous; papillae
present but not numerous, more or les uniform in size.
Short cells: present over the veins in rows of 3-5 or more cells. Pairs of short cells
occasionally found between the veins.
Prickle hair: none seen.
Micro hair: none seen.
Macro hair: none seen.
Stomata: large, numerous stomata; 2-3 banded between the veins; dome shaped stoma
present; paracytic stomata type seen.
Anticlinal wall: wavy.
Silica bodies: seen; mostly cross shaped

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Figure 2 b. The leaf epidermal of Cymbopogon giganteus. Adaxial epidermis;

sc short cell; lc long cell; s stomata; sb silica body; p papillae

DISCUSSION

The leaf epidermal layers of Cymbopogon citratus and Cymbopogon giganteus

showed significant degrees of variations in terms of the anatomical characters found in them.
Metcalfe (1954) pointed out that certain characters of the epidermis such as shape of the
subsidiary cells of the stomata, micro hairs and silica bodies are important systematically. Of
all the epidermal characters studied in the two species, the length of long cells, the micro hairs
and prickle hairs showed significant differences between Cymbopogon citratus and
Cymbopogon giganteus. The long cells found in the epidermis of Cymbopogon citratus were
longer than those found in the epidermis of Cymbopogon giganteus. Papillae were also seen
alongside the long cells of Cymbopogon giganteus while they were absent in Cymbopogon
citratus. The long cells found in Cymbopogon citratus were 5-7 rowed between the veins on
the abaxial and 8-10 rowed between the veins on the adaxial surface while the long cells
found in Cymbopogon giganteus were 4-7 rowed and 9-11 rowed between the veins on the
abaxial and adaxial epidermis respectively. The characters of diagnostic importance in the
identification of Cymbopogon citratus are micro hairs, which were sparsely distributed, in the
adaxial epidermis and prickle hairs present in both abaxial and adaxial epidermis. Micro hairs
were seen sparsely distributed in the adaxial epidermis of Cymbopogon citratus while they
were absent in both the abaxial and adaxial epidermis of Cymbopogon giganteus. This
anatomical character clearly separates the two species of Cymbopogon and could be employed
in characterizing them. The diagnostic characters for Cymbopogon giganteus are the papillae
which were absent in C. citratus. Metcalfe and Chalk (1979) reported that papillae
concentrates the limited light below the canopy of the tropical rain forests by acting as lenses
and also, facilitates the removal of water from the leaf surface.
In addition, abundance of stomata was observed on the adaxial epidermis of the two

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species while stomata were found sparsely distributed on the abaxial epidermis of both
species. But the arrangement of the stomata still showed some differences in that, the stomata
present were 2-banded between the veins on the abaxial and 2-4 banded between the veins on
the adaxial epidermis of Cymbopogon citratus. In Cymbopogon giganteus, the stomata were
seen 2-bandeed between the veins on the abaxial and 2-3 banded between the veins on the
adaxial epidermis. Due to the close relationships between the two species, some characters
such as stoma length and width, shapes of the subsidiary cells, silica bodies and cork cells
seen among the accessions of the two species studied could not adequately distinguish
between Cymbopogon citratus and Cymbopogon giganteus. Although the length of stoma
showed slight differences, the width of stoma found in the two species were almost the same.
In addition, the average widths of long cells seen in the adaxial epidermis of the two species
were more or less the same.
The present study reveals that some of the characters, which were present in the two
species, may be typical of the genus despite the many anatomical variations between them.
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citratus stapf grown in Zambia, Flavour and Fragrance Journal, 13, 1, 29-30.
2. Cruz, M. E. S., 2002, Allelopathic effect of Cymbopogan citratus and Artemisia absinthium on seeds of
Bidens pilosa, Acta Horticulturae 569, 229-233.
3. Cutler, D. F., 1978, Applied Plant Anatomy, 1st ed. Longman Incorporation .New York, 5-6.
4. Hutchinson, J., J. M. Dalziel, 1972, Flora of West Tropical Africa, Vol. III, Part. 2, Whtefriars Press,
Revised by Crown Agents, London, 459-461.
5. Metcalfe, C. R., L. Chalk, 1950, Anatomy of the dicotyledons, ed.1, vol. 1, Clarendon Press, Oxford
222-234.
6. Metcalfe, C. R., L. Chalk, 1979, Anatomy of the dicotyledons, 2nd ed. vol. 1, Clarendon Press, Oxford,
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7. Metcalfe, C. R, 1954, Recent work on the systematic anatomy of the Monocotyledons (with special
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A.Rich.) Stapf and Brachiaria callopus (pilg.) Stapf (poeaceae), Feddes Repertorium, 102, 3-4, 159-166.
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Indian Journal of Agronomy, 25, 1, 42-44.
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REZUMAT
STUDII COMPARATIVE ALE EPIDERMEI FOLIARE LA CYMBOPOGON CITRATUS
(STAPF.) I LA CYMBOPOGON GIGANTEUS (HOCHST.) CHIOV. N NIGERIA
Studiile foliare epidermale au fost realizate la speciile Cymbopogon citrates i Cymbopogon
giganteus cu scopul de a determina variaiile paternale la caracterele epidermale ale frunzelor i
aprecierea valorilor unor indici n identificarea i clasificarea speciilor. Dintre caracterele cu
importan de diagnostic, periorii au o distribuie haotic n epiderma inferioar, iar periorii
neptori sunt prezeni att n epiderma superioar, ct i n epiderma inferioar a frunzelor.
Caracterul de diagnostic pentru C. giganteus sunt papilele care se pot observa de-a lungul celulelor
alungite ale frunzelor.

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