Corresponding Author: Novi Dwi Octavia
E-mail: [email protected]
INTRODUCTION
Mentha is a genus in the Lamiaceae family with
approximately 30 species. Mentha is widely cultivated on five continents,
especially in temperate and subtropical areas, and is classified into five
genera containing 25-30 species (Chen et al., 2012). Several Mentha species include M. arvensis, M. aquatic, M. canadines, M.
pulegium, M. x piperita, M. piperita, and M. spicata. Among the Mentha species,
M. x piperita, M. piperita, and M. spicata are the most common in Indonesia.
Several Mentha species have
been successfully developed in 25 species. Still, there are only four Mentha
species that can be cultivated for essential oil production in the world,
namely M. canadensis, M. gracilis, M. piperita, and M. spicata (Chen et al.,
2012 ). Essential oils are aromatic
oil liquids that can be extracted from various parts of plants, namely leaves,
seeds, flowers, bark, and fruit skin (Tongkuanchan
& Benjakul, 2014). Mentha contains essential
oils, namely many chemicals in aromas such as menthol, menthone, iso menthone,
and menthofuran (Hidayat
& Rurini Retnowati, 2013).
M. piperita and M. spicata are
known for their essential oil content produced from trichome glands and also
for the abundance of phenolic compounds in the leaves (Dhifi et
al., 2016) ; (Kalemba
& Synowiec, 2019) ; (Park et al.,
2019) ; (Park et al.,
2019). Mentha species have various
types of trichomes, generally glands found in the epidermis and containing
essential oils. M. piperita can produce an essential oil commonly called
peppermint, while M. spicata can produce an essential oil commonly called
spearmint. The essential oil obtained from Mentha species is used extensively
in the pharmaceutical industry. Mentha species have antioxidant and
antimicrobial activity and the most active compounds that play a role are
menthol (Ali et al., 2015) ; (Mahdavikia
& Saharkhiz, 2015).
Based on previous research on the species M. piperita
and M. spicata, we examined several Mentha species, including M. piperita and
M. spicata, which included classification of inflorescence characters,
classification of leaf characters, taxonomic identification, and showing the
presence of trichome types in several species. Mentha (�arić-Kundalić
et al., 2016) . Previous research also examined the amount of oil
content or yield produced by the M. piperita and M. spicata species leaves. The
results were linked to observations of the anatomy of the leaves of the oil
glands of the M. piperita and M. spicata species (Puspitasari et al . , 2021).
Research can also be a research support for observing
the anatomical structure of the leaves and oil glands of the M. piperita and M.
spicata plants that will be studied. However, in the previous study, samples of
plant species used came from different places from this study; the species to
be used came from Sidomulyo Village, Kec. Batu, Malang (�arić-Kundalić
et al., 2016) ; (Puspitasari et
al., 2021) .
One of the characteristic parts
of the leaf is the leaf epidermis tissue and its derivatives. Research on the
characteristics of the anatomical structures and oil glands in the leaves of M.
piperita and M. spicata is important because there is little information about
the structural characteristics of M. piperita and M. spicata in Indonesia.
Therefore, based on the above
background, this research was carried out, which aims to analyze the
characteristics of variations in the anatomical structure and oil glands in the
leaves of two Mentha species, which are related to the presence of essential
oils. The research results will be useful in obtaining information about the
diversity of anatomical structures and supporting data for Mentha species.
������������������������������������������������������������������������������������������������������������������������������������
METHOD
This type of
research is descriptive qualitative, namely analyzing the anatomical and oil
gland structure characteristics of the leaves of M. piperita and M. spicata species. This research was conducted for 4 months, December 2022 � April
2023. Plant sampling was carried out in Sidomulyo Village, Batu, Malang. The leaves' anatomical structure was
observed at the Microtechnical Laboratory, Department of Biology, Faculty of
Mathematics and Natural Sciences, Surabaya State University.
Figure
1. Map
The plant samples used were taken
fresh. Plant sample collection was carried out by randomly sampling 3 specimens
for each Mentha species. The samples taken are leaves from nodes 3-5,
which are free of disease and other materials. The preparations were made using
three methods: the fresh and wholemount incision method for longitudinal
and the paraffin method for transverse incision. Observation of the transverse anatomical structure of
leaves using materials including FAA (formalin acetic acid-alcohol) alcohol
graded 50%, 70%, 95%, and 100%, alcohol solution: xylol (3:1), alcohol
solution: xylol (1: 1), alcohol solution: xylol (1:3, xylol absolute 1 and
xylol absolute II, safranin 1%,
fast green 0.5% and entel l an, while for observation of the longitudinal anatomical structure
of the leaves using the appropriate
wholemount method. Using distilled water, kloroks , safranin 0.1 %, glycerin
30%.
The preparations were observed with a light microscope with a magnification of
40x10 with a scale of 0.19625/mm 2 (Marantika
et al., 2021).
The data were analyzed
descriptively and qualitatively using the two Mentha species' anatomical
structures and oil glands. Data analysis includes qualitative and
quantitative data. Qualitative data were obtained by means of a literature
review and similar research results, while the application of image j processed
quantitative data. The results of data analysis are then arranged in tables and
figures to make it easier to characterize each species.
The anatomical characteristics
observed in the longitudinal incisions were the structure of the epidermis, the
shape of the epidermis, the composition of the epidermal cells and their
derivatives, the number of oil glands, the shape of the oil glands, the size of
the oil glands and the density of the oil glands. The anatomical characters of
the leaves observed in the cross-section were leaf tissue, including adaxial
and abaxial epidermal cells, palisade parenchyma and spongy parenchyma, various
forms of glandular and non-glandular trichomes, shape of essential oil, size of
essential oil, distribution of essential oil, location essential oils in the
leaves of the M. piperita and M. spicata species respectively.
RESULTS AND DISCUSSION
The results showed that there were variations in the
structure of the epidermis and its derivatives and also found the presence of
essential oil glands in the leaves of M. piperita and M. spicata species. Obtained from Sidomulyo Village,
Batu, Malang. Leaf anatomy was observed by making transverse and longitudinal incisions
to determine variations in leaf anatomy and characteristics that indicate the
presence of oil glands.
Anatomical
Structure
Mentha species, the
results obtained were the presence of constituent tissues. Based on Figure 1,
it is known that there are several similarities and differences in the
characteristics of the two species. The anatomical characteristics of the
leaves shared by both species with a magnification of 40x10 and a field of view
scale of 0.19625/mm 2 are the presence of epidermal cells, palisade
parenchyma tissue, sponge parenchyma tissue, and trichomes. Each species has
one layer of leaf epidermal cells located on the adaxial and abaxial sides of
the leaf. The palisade parenchyma and spongy parenchyma in both species have
the same structure; namely, the palisade parenchyma looks neatly arranged under
the adaxial epidermal layer, while the spongy parenchyma shows an irregular cell
arrangement as well as different shapes and sizes above the abaxial epidermal
layer. The transverse anatomical
structure of the leaves in three Mentha species can be seen in Figure 1.
Figure 2.
Structure of cross sections of two Mentha species with a magnification
of 40x10; a) M. piperita, b) M. spicata ; (EAD): Adaxial
epidermis, (PP): Palisade parenchyma, (PS): Sponge parenchyma, (MA): Essential
oils, (TK): Trichomes, (EAB): Abaxial epidermis.
Observations
obtained in longitudinal sections of the two Mentha species with a
magnification of 40x10 and a field of view scale of 0.19625/mm 2 include
The shape of the epidermal cell walls and epidermal derivatives, namely stomata
and trichomes, also found the presence of essential oils, but in Figure 2 only
shows the presence of oil glands in M. spicata. The anatomical structure
of M. piperita and M. spicata leaves showed that the epidermis surface showed grooved
and irregular anticlinal walls. Another difference is showing the presence of
trichomes in Figure 2, which are only found in M. spicata. The results of observations on the type of stomata of the leaves of the
two Mentha species have the same type, namely the anomocytic type in
Figure 2. This can be seen from the sketch of the stomata in Figure 4.2. It can
be seen that the number and arrangement of guard cells in the stomata consists
of 1 guard cell.
Figure 3. Leaf anatomical structure of longitudinal sections of two Mentha
species with a magnification of 40x10; a) M. piperita, b) M.
spicata ; (black arrow: cell wall, blue arrow: stomata, red arrow:
trichomes, green arrow: essential oil).
Trichomes
The results of observations of
other anatomical structures in the leaves of two Mentha species are the
presence of epidermal derivatives in trichomes. Trichomes on M. piperita leaves are found in the abaxial epidermis,
while trichomes on M. spicata leaves are found in both adaxial and
abaxial epidermis, Table 1.
Table 1. Characteristics of
trichomes on two types of leaf species: M. piperita and M. spicata
|
A. M.
piperita species
|
|
Type
|
Structure
|
Basal Cells
|
location
|
Size (�m)
|
Picture
|
|
Glandular
(Unicellular)
|
Round blunt stalk tip
|
4
|
Leaf adaxial
|
36.362 �m
|
|
|
Glandular
(Unicellular)
|
The pointed end of the stalk
|
4
|
Leaf
adaxial
|
12.105 �m
|
|
|
Peltate
glandular
(Unicellular)
|
1 blunt stalk cell
|
6
|
Leaf adaxial
|
32.146 �m
|
|
|
Capitate
glandular
(Multicellular)
|
1 stalk cell, 1
round head cell
|
2
|
Abaxial
leaves
|
27.296 �m
|
|
|
Non-Glandular
(Multicellular)
|
1 stalk cell, 1
blunt head cell
|
6
|
Leaf adaxial
|
19,999 �m
|
|
|
Non-Glandular
(Unicellular)
|
1 cell is flat
|
2
|
Leaf
adaxial
|
54.677 �m
|
|
|
B. Species M. spicata
|
|
Type
|
Structure
|
Basal Cells
|
location
|
Size (�m)
|
Picture
|
|
Glandular
(Unicellular)
|
1 blunt stalk cell
|
2
|
Abaxial leaves
|
37.332 �m
|
|
|
Non-Glandular
(Multicellular)
|
2 stalk cells, 1 tapered head cell
|
6
|
Leaf
adaxial
|
53.778 �m
|
|
|
Non-Glandular
(Unicellular)
|
The tip of the stem is tapered
|
2
|
Abaxial leaves
|
87.566 �m
|
|
|
Non-Glandular
(Multicellular)
|
1 stalk cell, 1
elongated pointed head cell
|
4
|
Abaxial
leaves
|
203.812 �m
|
|
|
Non-Glandular
(Unicellular)
|
1 round stalk cell
|
4
|
Abaxial leaves
|
39.992 �m
|
|
|
Non-Glandular
(Unicellular)
|
1 blunt stalk cell
|
4
|
Abaxial
leaves
|
125.479 �m
|
|
|
Dendroid
|
1 long stalk cell
|
4
|
Abaxial leaves
|
∞
|
 
|
Of
trichomes found on the abaxial epidermis of M. piperita leaves, namely
glandular trichomes (secretions) and non-glandular trichomes (non-secretions)
as presented in Table 1. An abaxial transverse section of M. piperita found:
1) glandular trichomes unicellular, blunt rounded stalk tip structure, 4 basal
cells, located at the adaxial side of the leaf, size 36.362 �m; 2) unicellular glandular trichomes, rounded structures, and
pointed stalk ends, 4 basal cells, located at the adaxial side of the leaf, 12.105 �m in size; 3) unicellular plate glandular trichomes, the structure
of 1 blunt stem cell, 6 basal cells, located in adaxial leaf, size 32.146 �m; 4) multicellular capitate glandular trichome, structure of 1
stalk cell, 1 round head cell, 2 basal cells, located abaxially on the leaf,
size 27.296 �m; 5)
multicellular non-glandular trichomes, structure of 1 blunt stalk cell, 6 basal
cells, located in adaxial leaf, size 19.999 �m; 6)
unicellular non-glandular trichomes, the structure of 1 flat stalk cell, 2
basal cells, located in adaxial leaves, size 54.677 �m.
Types of trichomes found in the abaxial and adaxial epidermis of M.
spicata plants, namely glandular trichomes and non-glandular trichomes as
presented in (Table 4.1). An abaxial transverse section of M. spicata found:
1) unicellular glandular trichomes, the structure of 1 blunt stalk cell, 2
basal cells, located abaxially on the leaf, size 37.332 �m; 2) unicellular non-glandular trichomes, tapered stalk tip
structure, 2 basal cells, located abaxially on the leaf, size 87.566 �m; 3) multicellular non-glandular trichomes, the structure of 1
stalk cell, 1 elongated pointed head cell, 4 basal cells, located abaxially on
the leaf, 203.812 �m; 4)
unicellular non-glandular trichomes, the structure of 1 round stalk cell, 4
basal cells, located abaxial in the leaf, size 39.992�m; 5) unicellular
non-glandular trichomes, structure of 1 blunt stalk cell, 4 basal cells,
located abaxially on the leaf, size 125.479 �m; 6)
unicellular non-glandular trichomes, structure of 1 round stalk cell, 4 basal
cells, located abaxially on the leaf, size 39.992 �m; 7) dendroid trichomes
with a structure of 1 cell on a long stalk, 4 basal cells, located abaxially on
the leaf. An adaxial longitudinal section of M. spicata found 1)
multicellular non-glandular trichomes, the structure of 2 stalk cells, 1
tapered head cell, and 6 basal cells, located abaxially on the leaf, size
53.778 �m (Table 1).
Oil Glands
Based on observations of the structure of oil gland cells in
the leaves of two Mentha species with
a magnification of 40x10 and a field of view scale of 0.19625/mm2, which has
similarities and differences in the number, shape, distribution, and
location of cells. The oil glands in M. piperita are located in the
lower epidermis. They are also spread across the palisade parenchyma cells and
sponge parenchyma, while the oil gland cells in M. spicata are located
in the upper epidermis (Figure 3).
Table 2. Results of observations on the amount, shape,
distribution, and location of oil in Mentha species
|
Species Name
|
Amount
|
Form
|
Distribution
|
Size
(�m)
|
location
|
|
M. piperita
|
25-35
|
Round
|
Spread
|
3,519 �m-8,642 �m
|
Located
in palisade parenchyma and spongy parenchyma
|
|
20-25
|
Round
|
Huddle
|
13,604 �m-24,012
�m
|
Located in the abaxial epidermis
|
|
M. spicata
|
10-15
|
Round
|
Huddle
|
38,986 �m-11,418
�m
|
Located in the adaxial epidermis
|
The characteristics of the oil glands with transverse
sections on the 2 leaf species of M. piperita and M. spicata showed
that in M. piperita, 1) oil glands number 25-35, have a small round
shape, are spread out and are located in the palisade parenchyma cells and
spongy parenchyma, have a size of 3.519 �m-8.642
�m; 2) oil glands number 20-25, have a large round shape, are
clustered in distribution and are located in the lower epidermis, have a size
of 13.604 �m-24.012 �m; whereas in M. spicata; 1) Oil glands number 10-15, have a large
round shape, are clustered in distribution and are located in the upper
epidermis, have a size of 38.986 �m-11.418 �m (figure 3).
Figure 4. Images
of the oil glands of two Mentha species: a) M. piperita,
M. spicata;
black arrow: location of oil glands
The
results of anatomical observations of longitudinal incisions in the leaf
epidermis of the two Mentha species with a magnification of 40x10 and a
field of view scale of 0.19625/mm 2 show that the two species have
the same epidermal cell shape Figure 4.1. Epidermal cells of Mentha
leaves are curved, irregular, and have different cell sizes. There are
variations in the characteristics of the oil glands, namely in the size and
number of the epidermis, Table 3.
Table 3.
Number, shape, and size of oil gland cells in
leaves of two
species M. piperita and M. spicata
|
Species Name
|
Amount
|
Form
|
Size
|
|
M. piperita
|
698
|
Round
|
3.124 �m - 6.719 �m
|
|
M. spicata
|
482
|
Round
|
2.112 um 3.734 �m
|
Mentha species in a longitudinal section with a magnification of 40x10 and a field of view scale of
0.19625/mm 2 can be seen in Figure 4.4. It can be
seen in the picture that there are round characters that are inside the
epidermal cells and have a round shape and also spread. Some of these oil glands
are yellow and clear (Figure 4.4). The number of oil glands in Figure 4.4 is
that in the species M. Piperita has a total of 698 and has a size of 3.124 �m -6.719 �m while in the species M. spicata, there
are 482 and a size of 2.112um-3.734 �m
(table 3)
Figure 5. Image of sebaceous gland cell density in incision
longitudinal in two Mentha species ; a) M. piperita , b) M.
spicata
Plants
from the Mentha clan, namely M. piperita and M. spicata,
can produce essential oils. Essential oils or essential oils are liquid oils
extracted from plant parts, namely leaves, seeds, flowers, bark, and fruit
peels (Tongkuanchan &
Benjakul, 2014). Several food and pharmaceutical industries widely use the essential oil of the Mentha clan because the plant
has antioxidant and antimicrobial activity (Ali et al., 2015) ; (Mahdavikia & Saharkhiz, 2015).
In this study, observations were made of the
anatomical structure of the leaves of two Mentha species in Sidomulyo
Village, Batu, Malang, namely M. piperita and M. spicata, by
making transverse and longitudinal incision preparations with a microscope
magnification of 40x10 magnification and a field of view scale of 0.19625/mm2
which was carried out based on preliminary tests and previous research
references which are useful for knowing variations in leaf anatomy between the Mentha
species studied and the anatomical characteristics of leaves that have oil
glands (�arić-Kundalić et al., 2016).
Based on the results of observations of
cross-sections on the leaves of the two Mentha species, they have one
thing in common, one of which is a thick, unstained epidermal layer, because
this tissue does not absorb fast green dye during the staining process. Neatly
arranged palisade parenchyma beneath the adaxial epidermis and irregularly
shaped spongy parenchyma. The parenchyma in both Mentha species is
included in the assimilation parenchyma because there are two types of
assimilation parenchyma: palisade and spongy (Palennari et al., 2016). Palisade
parenchyma and spongy parenchyma belong to the mesophyll structure of the
leaves of two Mentha species, which function in photosynthesis. This is
supported by other research stating that the palisade parenchyma consists of
several layers of columnar cells with many chloroplasts. At the same time, the
sponge parenchyma consists of 4-6 layers of cells with chloroplastids and
irregular shapes and air spaces between cells where these chloroplasts function
in the process. Photosynthesis (Rita & Animesh, 2011).
Based on the results of observations of longitudinal incisions
that were carried out on the leaves of two Mentha species, namely M.
piperita and M. spicata, there were results that the epidermal cell
walls were in the form of polygonal anticlinals, namely grooved and curved. The
cell wall can be curved if indicated without the formation of corners and has
inward and outward curves (Febriyani et al., 2022). There are
differences in the shape of the epidermal cells, but the epidermis is a uniform
tissue (Anu et al., 2017). The
similarity in the shape of the epidermal cell walls in the two Mentha
species is because both belong to the same genus, namely Mentha. The
Mentha plant is a genus of the Lamiaceae family, which has approximately
30 species and various hybrids and generally grows in sub-tropical areas
(Chen et al., 2012). Among the
several species of the Mentha plant, Mentha x piperita, M. piperita, and M. spicata are the most
common in Indonesia.
The results of observations on the type of
leaf stomata of both Mentha species have the same type, namely the anomocytic
type. The type of stomata is anomocytic; the guard cells are surrounded by
several cells that are the same shape and size as other epidermal cells (Fauziah & Izzah,
2019). These two Mentha species have
little or no stomata, and this is following research (Rita & Animesh,
2011). The similarity in the number of stomata
in the two Mentha species is caused by data collection carried out in
the same area so that more or less environmental factors in light intensity,
temperature, and humidity do not have much difference. Previous research stated
that stomata density can also be influenced by environmental factors, namely
temperature, light intensity, and humidity (Sundari & Atmaja,
2017). The number of stomata on the leaves of M.
piperita and M. spicata is small because it is influenced by the
environment, where it is known that the habitat of M. piperita and M.
spicata is in cool to cold places, one of which is located in Sidomulyo
Village, Batu, Malang. is located at the foot of a mountain to the south of
Mount Arjuna and has an altitude of 800�850 meters above sea level,
with an air temperature of 17oC� 25oC and fertile soil conditions (Ningtias, 2022). The number of stomata tends to decrease
in an environment with low temperature and high humidity (Paluvi & Mukarlina,
2015).
Trichomes are found on both sides of the
leaf, both on the adaxial and abaxial sides in M. spicata, while in M.
piperita, the trichomes are only found on the abaxial side. This aligns
with previous research that M. spicata has trichomes on both sides of
its leaves (�arić-Kundalić
et al., 2016). The types of trichomes found in both
species consist of glandular trichomes and non-glandular trichomes. Glandular
trichomes are trichomes capable of producing secretions, while non-glandular
trichomes are trichomes that do not. The types and forms of non-glandular
trichomes are scale hairs, branched (multi-celled) hairs, and single hairs,
while the types and forms of glandular trichomes are trichomes, salt glands,
oil glands, and itchy hairs. The structure and morphology of trichomes are
diverse and can be used as a key to identifying genera, species, subspecies,
and varieties from various families (Harisha & Jani,
2013).
Types of trichomes in M. spicata are
glandular and non-glandular in the abaxial and adaxial epidermis; they consist
of unicellular glandular trichomes as well as unicellular and multicellular
non-glandular trichomes. M. Piperita has a glandular, glandular planet,
glandular capitate, and non-glandular trichomes, which are found only in the
abaxial epidermis; consists of unicellular glandular trichomes, unicellular
plate glandular trichomes, unicellular capitate glandular trichomes and
unicellular and multicellular non-glandular trichomes (table 4.1). Unicellular
trichomes consist of one constituent cell, while multicellular trichomes
consist of more than one constituent cell (Yuliani & Ratnawati,
2018). Variations in the cells make up the
trichomes in M. piperita and M. spicata, including the shape and
number of basal cells, stalk cells, and head cells.
M. piperita shows the presence of glandular peltate and glandular capitate
trichomes. In the Lamiaceae family, two types of glandular trichomes are found:
peltate and capitate. In contrast to M. spicata, where no glandular peltate and
glandular capitate trichomes were found, this was because the leaf samples used
were not young leaves but leaves at nodes 3-5. The density of glandular
trichomes in Lamiaceae will decrease as the organ ages, and the peltate and
capitate trichomes will experience damage to the head cell sheath when entering
the aging phase of the secretory structure along with the aging of the leaf
organs. The trichomes found in plants of the Coleus genus, which are members
of the Lamiaceae, show morphologically, the head cells of pelvic and capitate
glandular trichomes in the three species from the visible presence of essential
oils (essential oils) and other secondary metabolites which experience an
increase in production at the 1st node and decrease at the nodes 3 and 5. Plant
adaptation mechanisms because leaves in the early growth phase require higher
protection to protect themselves from predators. Secretory structures are
generally capable of secreting substances that are toxic to herbivores. Peltate
glandular trichomes that accumulate monoterpenes. These trichomes contain
secretory cells responsible for oil synthesis (Rios-Estepa et al., 2010). The
difference that the M. spicata species has and that is not found in M.
piperita is the presence of dendroid trichomes (branched trichomes). This
follows previous research that branched trichomes (dendroid trichomes) are only
found in a few Mentha genus species and not in all types (�arić-Kundalić et al., 2016).
Previous research stated that the formation of trichomes begins
with a bulge in the epidermal cells. The bulge can enlarge and grow into
multicells, and then various shape modifications occur following growth (Kurrataa'yun, 2013). Mentha species,
especially parenchyma cells, will undergo a process of proliferation and
elongation so that stalk cells develop and glandular trichomes can form. This
is in accordance with the statement that essential oils are produced in
secretory cells initially originating from parenchymal cells undergoing a
differentiation process (Dorly et al., 2015). The phase of trichome formation can be called the pre-secretory
phase. The glandular trichomes develop and become complete structures
consisting of basal, stalk, and secretory cells. The development phase can
continue in the secretory phase; namely, the glandular trichomes can accumulate secondary metabolites optimally, which is characterized
by a gradual increase in size.
Secretory cells are special secretory structures that can secrete
certain compounds where these compounds are not excreted by plants (Nindyawati & Indriyani, 2017). The secretion results through the secretory are in the form of oil,
resin, latex, mineral salts, and various other chemical compounds. Secretory
structures can take the form of idioblast cells, secretory cavities, glandular
trichomes, non-glandular trichomes, laticifers, resin ducts, and oil glands (Kuster & Vale, 2016) (de Almeida et al., 2020). Secretory cells have a location that can
be divided into two, namely internal and external secretory. The internal
secretaries are in the form of idioblasts, secretory cavities, secretory ducts,
and laticifers, while the external secretaries are in the form of trichomes,
nectaries, stigmas, and hydathodes (Dorly et al., 2015). Mentha essential oil species
found were in two secretory places, namely the internal and external
secretories. The internal secretory of essential oils is in idioblast cells.
Idioblasts can be found in the mesophyll area, which can be in the form of cell
spaces or oil glands, mucus cells, or crystal cells. The external secretory of
essential oils is in glandular trichomes. Trichomes in the genus Mentha
function as storage and as synthesis in the oil glands, following Lange's
statement that the essential oils of Mentha plants are synthesized and
stored in trichomes, namely in the modified epidermis, and are called glandular
trichomes. Mentha plant trichomes contain secretory cells which are responsible for
oil synthesis. Gershenzon and McCaskill supported that Mentha's biosynthetic
processes and accumulation of monoterpenes were specifically localized to the
glandular trichomes.
The results of anatomical observations of
cross-sectional sections of M. piperita and M. spicata leaves showed
the structure of essential oil glands, which had a round and brownish-yellow
color. The structure of the oil glands in both species varies in number, shape,
distribution, and location of cells. Oil gland cells in M. piperita are distributed
in the palisade and spongy parenchyma and cluster in the abaxial epidermis. In
contrast, the oil gland cells in M. spicata are distributed in the
adaxial epidermis (Figure 4.3). This is also supported by the results of
longitudinal incisions in the adaxial epidermis of the two Mentha species, showing
the presence of oil glands that appear only in M. spicata species (Figure
4.2).
The number of oil gland
cells in the M. piperita leaf species shows that it has 20-35 oil gland
cells with details of 25-35 oil gland cells in the abaxial epidermis, which has
a size of 13.604 �m-24.012�m and 20-25 oil gland cells are spread out in the palisade parenchyma and spongy
parenchyma which have a size of 3.519 �m-8.642�m while M. spicata has
several oil glands of 10-15 cells which are on the abaxial side which has a
size of 38.986 �m-11.418�m. This
certainly affects the oil content results in each Mentha species, where
the more oil gland cells, the more oil content will be produced. According to
Atmono's statement, the increase in secretory cells aligns with cell division.
This is also in line with other studies on the maceration test using 96% pa
ethanol solvent with each species using 50g of simplicia powder; the results of
the study stated that the M. piperita species obtained more oil yield,
namely 70.4%, whereas in M. spicata obtained less oil yield, namely 69% (Puspitasari et al., 2021).
The shape
of the oil gland cells in the leaves of M. piperita and M. spicata, as
seen in longitudinal sections, is round and spread out (Figure 4.4). There are
variations in the characteristics of oil glands, namely in size and number in
one field of view. M. Piperita has more oil gland cells than M.
spicata, namely 698, while M. spicata has several oil gland cells,
482. There is a relationship between the density of gland trichomes and the
results of essential oil content (Gupta et al., 2017), (Mishra et al., 2018). The diameter of the M. piperita oil
gland cells is larger than that of M. spicata, namely 3.124 �m - 6.719
�m, while M. spicata has an oil gland cell diameter of 2.112 �m - 3.734
�m. The increase in size of secretory
cells containing essential oil glands can be in line with growth, which
includes the process of cell and tissue expansion. The greater the
density of the oil glands and the larger the size of the oil glands, the more
essential oil can be produced.
Species
from the Mentha genus are known to have medicinal and commercial
importance (Chen et al., 2012) ; (Jeyakumar et al., 2011). Previous
research stated that the M. piperita species had the highest total phenolic
content, followed by M. spicata (Zaidi & Dahiya, 2015). This is also supported by (Gharib & da Silva, 2013) ; (Naidu et al., 2012). Furthermore, in other research, phytochemical
screening tests were obtained and the results showed that two Mentha species
contained flavonoids, steroids, saponins, and tannins (Puspitasari et al., 2021). This is also supported by previous research on
phytochemical screening tests showing the presence of various bioactive
substances such as phenols and tannins, flavonoids, glycosides, and alkaloids
in methanol (Khanal, 2020). Mentha's antioxidant properties come from the content of active components such as
menthone, menthol, rosmarinic acid, and carvone (Lawrence,
2013).
Other
previous studies stated that the essential oil content of M. piperita and
M. spicata species found antimicrobial activity against clinical
isolates, phytochemicals, phenolic content, and bioactive compounds, which are
responsible for antibacterial potential by TLC ( Thin-Layer Chromatography )
bioautographic analysis (Zaidi & Dahiya, 2015). Several
previous studies also revealed that there was antibacterial activity in M.
piperita and M. spicata species, namely Staphylococcus aureus,
Escherichia coli, Klebsiella spp and also had antifungal activity
against Aspergillus spp . and Candida albicans (Sujana et al., 2013) ; (Jeyakumar et al., 2011) ; (CHAUHAN & AGARWAL, 2013). This
correlates with studies where the essential oil content of M. piperita and
M. spicata species showed antibacterial activity (Singh et al., 2015). Thus, the
species M. piperita and M. spicata can be said to contain
essential oils, which are a good source of natural antimicrobial agents.
CONCLUSION
Based on the results of
research on the anatomical structure of the leaves of two Mentha species, it
shows that M. piperita has a greater number of essential oil glands compared to
M. spicata. M. piperita in longitudinal section has 698 oil gland cells, while
M. spicata has 482 oil gland cells. M. piperita in transverse section has 20-35
oil gland cells, while M. spicata has 10-15 oil gland cells. The diameter of the
oil gland cells of M. piperita is larger than that of M. spicata, namely
3.124�m - 6.719�m, while M. spicata has an oil gland cell diameter of 2.112�m -
3.734�m. These results certainly affect the results of the essential oil
produced.
REFERENCES
Ali, B., Al-Wabel, N. A., Shams, S., Ahamad, A., Khan, S. A.,
& Anwar, F. (2015). Essential oils used in aromatherapy: A systemic review.
Asian Pacific Journal of Tropical Biomedicine, 5(8), 601�611.
Anu, O., Rampe, H. L., & Pelealu, J.
J. (2017). Struktur sel epidermis dan stomata daun beberapa tumbuhan suku
euphorbiaceae. Jurnal MIPA, 6(1), 69�73.
CHAUHAN, S. S., & AGARWAL, R. (2013). Evaluation of
antibacterial activity of volatile oil from Mentha spicata L. Journal of
Drug Delivery and Therapeutics, 3(4), 120�121.
Chen, X., Zhang, F., & Yao, L. (2012). Chloroplast DNA
molecular characterization and leaf volatiles analysis of mint (Mentha;
Lamiaceae) populations in China. Industrial Crops and Products, 37(1),
270�274.
de Almeida, V. P., Raman, V., Raeski, P. A., Urban, A. M.,
Swiech, J. N., Miguel, M. D., Farago, P. V., Khan, I. A., & Budel, J. M. (2020).
Anatomy, micromorphology, and histochemistry of leaves and stems of Cantinoa
althaeifolia (Lamiaceae). Microscopy Research and Technique, 83(5),
551�557.
Dhifi, W., Bellili, S., Jazi, S., Bahloul, N., & Mnif, W.
(2016). Essential oils� chemical characterization and investigation of some
biological activities: A critical review. Medicines, 3(4), 25.
Dorly, D., Wiryo, B. A., Nurfaizah, I., & Nidyasari, R.
R. S. (2015). Secretory structure and histochemistry tests of Asteraceae family
members of medicinal plants in Walat Mountain Educational Forest. Proceeding
Biology Education Conference: Biology, Science, Enviromental, and Learning,
12(1), 667�673.
Fauziah, A., & Izzah, A. S. Z. (2019). Analisis tipe
stomata pada daun tumbuhan menggunakan metode stomatal printing. Prosiding
Seminar Nasional Hayati, 7, 34�39.
Febriyani, H., Puspitawati, R. P., & Bashri, A. (2022).
Variasi Struktur Anatomi dan Sekretori Pada Spesies Annona Yang Berpotensi
Sebagai Tanaman Obat. LenteraBio: Berkala Ilmiah Biologi, 11(3),
575�585.
Gharib, F. A., & da Silva, J. A. T.
(2013). Composition, total phenolic
content and antioxidant activity of the essential oil of four Lamiaceae herbs. Medicinal
and Aromatic Plant Science and Biotechnology, 7(1), 19�27.
Gupta, S., Jain, U., & Chauhan, N. (2017). Laboratory
diagnosis of HbA1c: a review. J Nanomed Res, 5(4), 120.
Harisha, C. R., & Jani, S. (2013). Pharmacognostical
study on trichomes of Solanaceae and its significance. Univers J Pharm, 2,
100�104.
Hidayat, F., & Rurini Retnowati, S. (2013). Isolasi Dan
Karakterisasi Komponen Minyak Mintdari Daun Mentha Arvensislinn. Fakultas Matematika Dan Ilmu Pengetahuan
Alam. Universitas Brawijaya. Jl. Veteran Malang, 65145, 567�573.
Jeyakumar, E., Lawrence, R., & Pal, T. (2011).
Comparative evaluation in the efficacy of peppermint (Mentha piperita) oil with
standards antibiotics against selected bacterial pathogens. Asian Pacific
Journal of Tropical Biomedicine, 1(2), S253�S257.
Kalemba, D., & Synowiec, A. (2019). Agrobiological
interactions of essential oils of two menthol mints: Mentha piperita and Mentha
arvensis. Molecules, 25(1), 59.
Khanal, B. (2020). Phytochemical and antibacterial
analysis of Mentha piperita (Peppermint). Thesis-Dissertation, no.
Kurrataa�yun. (2013). Analisis struktur anatomi akar dan
batang pohon aren sebagai pohon yang dapat mencegah banjir dan erosi.
Institut Pertanian Bogor.
Kuster, V. C., & Vale, F. H. A. (2016). Leaf
histochemistry analysis of four medicinal species from Cerrado. Revista Brasileira
de Farmacognosia, 26, 673�678.
Lawrence, B. M. (2013). The story of India�s mint oils and
menthol. Perfumer Fla, 38, 26�35.
Mahdavikia, F., & Saharkhiz, M. J. (2015). Phytotoxic
activity of essential oil and water extract of peppermint (Mentha � piperita L.
CV. Mitcham). Journal of Applied Research on Medicinal and Aromatic Plants,
2(4), 146�153. https://doi.org/10.1016/j.jarmap.2015.09.003
Marantika, M., Hiariej, A., &
Sahertian, D. E. (2021). Kerapatan dan distribusi stomata daun spesies mangrove
di Desa Negeri Lama Kota Ambon. Jurnal
Ilmu Alam Dan Lingkungan, 12(1).
Mishra, A., Jain, P., Lal, R. K., & Dhawan, S. S. (2018).
Trichomes and yield traits in Mentha arvensis: Genotype performance and
stability evaluation. Journal of Herbs, Spices & Medicinal Plants, 24(1),
1�14.
Naidu, J. R., Ismail, R. B., Yeng, C., Sasidharan, S., &
Kumar, P. (2012). Chemical composition and antioxidant activity of the crude
methanolic extracts of Mentha spicata. Journal of Phytology, 4(1).
Nindyawati, D. L., & Indriyani, S. (2017). Struktur Sel
Sekretori dan Uji Mikroskopi Mikrokimiawi Metabolit Sekunder pada Daun dari
Tujuh Taksa Tanaman Obat Antihipertensi. Biotropika: Journal of Tropical
Biology, 5(2), 59�67.
Ningtias, R. A. (2022). Pelaksanaan Penyusunan Laporan
Penyelengaraan Pemerintahan Daerah (LPPD) Melalui Aplikasi E-LPPD Kota Batu
(Studi di Sekretaris Daerah bagian Pemerintahan dan Otonomi Daerah Kota Batu).
Universitas Muhammadiyah Malang.
Palennari, M., Lodang, H., Faisal, F., & Abd, M. (2016). Biologi
Dasar Bagian Pertama. Alauddin University Press.
Paluvi, N., & Mukarlina, R. L. (2015). Struktur Anatomi
Daun, Kantung dan Sulur Nepenthes gracilis Korth. yang Tumbuh di Area
Intensitas Cahaya Berbeda. Jurnal Protobiont, 4(1). http://dx.doi.org/10.26418/protobiont.v4i1.9452
Park, Y. J., Baek, S.-A., Choi, Y., Kim, J. K., & Park,
S. U. (2019). Metabolic profiling of nine Mentha species and prediction of
their antioxidant properties using chemometrics. Molecules, 24(2), 258. https://doi.org/10.3390/molecules24020258
Puspitasari, L., Mareta, S., &
Thalib, A. (2021). Karakterisasi Senyawa
Kimia Daun Mint (Mentha sp.) dengan Metode FTIR dan Kemometrik. Sainstech
Farma: Jurnal Ilmu Kefarmasian, 14(1), 5�11. https://doi.org/10.37277/sfj.v14i1.931
Rios-Estepa, R., Lange, I., Lee, J. M.,
& Lange, B. M. (2010). Mathematical
modeling-guided evaluation of biochemical, developmental, environmental, and
genotypic determinants of essential oil composition and yield in peppermint
leaves. Plant Physiology, 152(4), 2105�2119. https://doi.org/10.1104/pp.109.152256
Rita, P., & Animesh, D. K. (2011). An updated overview on
peppermint (Mentha piperita L.). International Research Journal of Pharmacy,
2(8), 1�10.
�arić-Kundalić, B., Fialov�, S., Dobe�, C., �lzant,
S., Tekeľov�, D., Grančai, D., Reznicek, G., & Saukel, J. (2016).
Addendum: �arić-Kundalić, B.; Fialov�, S.; Dobe�, C.; �lzant, S.;
Tekeľov�, D.; Grančai, D.; Reznicek, G.; Saukel, J. Multivariate
Numerical Taxonomy of Mentha Species, Hybrids, Varieties and Cultivars. Sci.
Pharm. 2009, 77, 851�876. Scientia Pharmaceutica, 84(4), 752.
Singh, R., Shushni, M. A. M., & Belkheir, A. (2015).
Antibacterial and antioxidant activities of Mentha piperita L. Arabian
Journal of Chemistry, 8(3), 322�328. https://doi.org/10.1016/j.arabjc.2011.01.019
Sujana, P., Sridhar, T. M., Josthna, P.,
& Naidu, C. V. (2013). Antibacterial
activity and phytochemical analysis of Mentha piperita L.(Peppermint)�An
important multipurpose medicinal plant.
Sundari, T., & Atmaja, R. P. (2017). Bentuk sel
epidermis, tipe dan indeks stomata 5 genotipe kedelai pada tingkat naungan
berbeda. Jurnal Biologi Indonesia, 7(1).
Tongnuanchan, P., & Benjakul, S. (2014). Essential oils:
extraction, bioactivities, and their uses for food preservation. Journal of
Food Science, 79(7), R1231�R1249. https://doi.org/10.1111/1750-3841.12492
Yuliani, E., & Ratnawati, R. (2018). Studi Keanekaragaman
Struktur Dan Kepadatan Trikoma Glanduler Pada Beberapa Tanaman Obat. Kingdom
(The Journal of Biological Studies), 7(4), 262�268.
Zaidi, S., & Dahiya, P. (2015). In vitro antimicrobial
activity, phytochemical analysis and total phenolic content of essential oil
from Mentha spicata and Mentha piperita. International Food Research Journal,
22(6), 2440.
