The structure of cystoliths in selected taxa of the genus Ficus L.

(Moraceae) in
Peninsular Malaysia
B. Ummu-Hani and T. Noraini

Citation: AIP Conference Proceedings 1571, 372 (2013); doi: 10.1063/1.4858686

View online: http://dx.doi.org/10.1063/1.4858686
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Published by the American Institute of Physics
 The Structure of Cystoliths in Selected Taxa of the Genus
Ficus L. (Moraceae) in Peninsular Malaysia
B. Ummu-Hani and T. Noraini

School of Environmental and Natural Resource Sciences, Faculty of Science and Technology,
Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor

Abstract. A study was undertaken on mature leaves of 15 taxa of the genus Ficus in Peninsular Malaysia. The main
objectives of this study are to determine the morphology and distribution of cystoliths in the epidermal layers of the leaf
lamina in selected taxa of Ficus. The morphology of cystoliths is classified based on its size, shape, colour, and the
presence of stalk cystolith. There are seven types of cystolith morphology observed in this study. Most of the cystoliths
are either solitary, elongated, narrow or broad, and pointed or blunt at one or both ends. However, double- and rarely
triple-cystoliths are also present in some species. The size of the cystoliths varies even within the same species. Based on
the position of cystoliths, all the 15 taxa studied can be generally classified into three groups: Group 1 - with cystoliths
adjacent to the adaxial epidermis layer (F. annulata, F. benghalensis and F. superba), Group 2 - with cystoliths adjacent
to the abaxial epidermis layer (F. aurantiacea, F. lepicarpa, F. hispida, F. obscura var. borneensis, F. religiosa, F.
schwarzii, F. ucinata and F. vasculosa), and Group 3 - with cystoliths present in both adaxial and abaxial epidermis
layers (F. benjamina, F. depressa, F. microcarpa and F. tinctoria). Based on the occurrence of cystoliths, the types of
lithocysts were related to the number of epidermal layers, i.e. hair-like lithocysts in uniseriate epidermis is present in all
species studied. However, the characteristics of the cystoliths may not suitably be used as a taxonomic marker but it can
be useful as additional character for group identification in Ficusper.
Keywords: Moraceae, Ficus, cystoliths, calcium carbonate, crystals.

INTRODUCTION
Ficus L. is the largest genus in Moraceae which comprises of 1,000 species worldwide, mostly found in tropical
and subtropical regions with up to 105 species in the African floristic region [1] and about 101 species recorded in
Peninsular Malaysia [2]. These species of trees, shrubs, climbers and hemi-epiphytic stranglers are recognized by a
specialised inflorescence and pollination syndrome [1].
Cystoliths are silicified bodies with cellulose skeleton or occasionally not encrusted. They are generally found
only in a few families such as Moraceae, Urticaceae and Acanthaceae [3]. The cystoliths can be identified either by
their nature, shape, size, colour and occurrence throughout the family. The cystoliths consisting of calcium
carbonate are usually located in the lithocysts. According to Mauseth [4], the lithocysts can be found in the form of
papillate or hair-like lithocysts and occur mostly in the epidermal layer of the leaves.
Since cystoliths are not considered common in occurrence or important for classification purposes, they are not
used as a diagnostic characteristic for a family, much less a genus. However, in some plant families such as
Moraceae, Acanthaceae, Urticaceae and Boraginaceae, various forms of cystoliths can be good criteria for genus and
species identification [5-6]. This type of cell inclusion has received attention of many plant anatomists such as
Metcalfe and Chalk [3], Solereder [5], Ahmad [7-9] and Inamdar et al. [10] Therefore, the purpose of this study are
to determine the morphology and distribution of cystoliths in the epidermal layers of the leaf lamina that may have
taxonomic value to be used as an identification characteristic of Ficus sp.

MATERIALS AND METHODS

A total of 15 taxa of Ficus were used in this study and the details of each species are given in TABLE (1). Fresh
specimens used in this study were obtained from several forest reserves in Selangor, Terengganu and Perak. Dried
specimens were obtained from Herbarium, Universiti Kebangsaan Malaysia (UKMB). Fresh leaves samples were
cut and fixed in AA solutions (70% Alcohol: 30% Acetic acid in a ratio of 1:3) for 48 hours before subjected to

The 2013 UKM FST Postgraduate Colloquium

AIP Conf. Proc. 1571, 372-376 (2014); doi: 10.1063/1.4858686
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372
further procedures. Lamina parts were sectioned (30m of thickness) using sliding microtome, stained in safranin
and alcian green, dehydrated in a series of ethanol with different concentrations (50%, 70%, 90%, 95 % and absolute
ethanol) and finally mounted on slides using Euparal, before kept in an oven at 50C for two weeks.
Photomicrographs of leaves sections were captured and processed using Cell^B software. Fixation and embedding
followed the methods described by Johansen [11] and Saas [12] with appropriate modifications. Description of data
was based on Metcalfe and Chalk [6], Solereder [5] and Ahmad [9].

RESULTS AND DISCUSSION

For convenience and to avoid lengthy and complex descriptions, the following illustrations of cystoliths
structures of transverse section of lamina have been described in TABLE (2), which are illustrated by
photomicrographs.
This study was mainly concerned with the presence of cystoliths, or calcium carbonate deposits in the leaf
epidermis. Sometimes cystoliths are found in the idioblasts cells [6]. According to Solereder [5], cystoliths also can
be found in the pith and cortex of plant stems. Amorphous cystoliths are found in several unrelated dicotyledonous
plant families and reported to be abundant in some members of the order Urticales, such as in Moraceae, Urticaceae,
and Ulmaceae [13].
The cystoliths is a spindle-shaped body composed of concentric layers of longitudinally orientated cellulose
microfibrils associated with pectins and other cell wall polysaccharides and at maturity it is heavily impregnated
with calcium carbonate [14]. Ahmad [9] mentioned the various shapes of cystoliths ie. round, oval, oblong, conical,
arc-shaped, bean-shaped, T, Y or V-shaped and for elongated cystoliths either spindle or cigar-shaped either with
blunt or pointed ends. Cylindrical, globular, bilanceolate and bilobed types have also been reported by Selvaraj and
Subramanian [15]. Metcalfe and Chalk [6] noted that true cystoliths known to occur in some genera in Moraceae
such as Broussonetia, Chlorophora, Conocephalus, Ficus, Morus, and so on. According to Esau [16] and Foster
[17], generally cystoliths in Ficus occur in the cells of the multiple epidermis. Various colours have been recorded
by Solereder [5] such as greenish, violet and blue green. Wu et al. [18] stated that, most lithocysts contain a stalk and
cystoliths body structure.
In the leaf lamina of 15 taxa of the genus Ficus studied, seven types of cystoliths morphology are observed. List
and descriptions of cystoliths structure found in this study are given in TABLE (2). Type 1 is present in F. annulata,
F. aurantiacea, F. benjamina, F. microcarpa, F. obscura var. borneensis and F. ucinata. The cystoliths adjacent to
the adaxial epidermis layer with oblong-shaped and blue colour. Type 2 also found in adjacent to the adaxial
epidermis layer but with cigar-shaped and violet colour. This type of cystoliths was observed only in F. religiosa.
Types 3, 4, 5, 6 and 7 are presented by cystoliths adjacent to the abaxial epidermis layer respectively but, they are
different in shape and colour and also the sizes of cystoliths and stalk. Type 3 is found in F. aurantiacea, F. obscura
var. borneensis, F. schwarzii, F. superba and F. ucinata with solitary rounded with blunt ends shaped and greenish
colour (Table 2). Type 4 is cystolith with a bean-shaped and blue colour which present in F. tinctoria only (TABLE
(2)). While, the cystolith with Y-shaped or double cystoliths and violet colour (Type 5) is found only in F.
vasculosa. Type 6 is only found in F. lepicarpa, the shape is solitary elongated cystolith with pointed ends and with
blue colour. Lastly, type 7 also is found only in F. hispida with solitary rounded-shaped and blue colour cystoliths.
The results have shown that all the cystolith shapes are composed of various sizes even in the same species but the
size for each species varies insignificantly.
Based on the position of their cystoliths, the 15 taxa studied can be classified into three groups, Group 1 with
cystoliths adjacent to the adaxial epidermis layer (F. annulata, F. benghalensis and F. superba), Group 2 with
cystoliths adjacent to the abaxial epidermis layer (F. aurantiacea, F. lepicarpa, F. hispida, F. obscura var.
borneensis, F. religiosa, F. schwarzii, F. ucinata and F. vasculosa) and, Group 3 with cystoliths present in both
adaxial and abaxial epidermis layer (F. benjamina, F. depressa, F. microcarpa and F. tinctoria,). A second and third
layer of epidermal-like cells was particularly well developed adaxially in all Ficus taxa studied. This layer is
referred as a hypodermis cells with highly thickened walls and reduced cytoplasmic contents at maturity similar to
the epidermal cells. Although the function of the hypodermis is apparently unknown, it may inrease the protective
and supportive function of the epidermis especially in marginal environment. This role is also related to some
function in adaptation to the surrounding habitat.
Lithocysts is a cell that contains cystoliths and stalk. According to Wu and Kuo-Huang [19], two types of
lithocysts are usually found in some Moraceae species, the hair-like lithocysts in uniseriate epidermis and papillate

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lithocysts in multiseriate epidermis. In this study, only hair-like lithocysts is observed in all Ficus taxa studied.
Furthermore, the results have shown that the presence of stalk in the lithocyts may have significance value
especially in providing additional data for species identification. Most of lithocysts found in Ficus species studied
contain stalk in various sizes except in F. religiosa and F. tinctoria, which are similar.
As a conclusion, a combination of cystoliths morphology such as size, shape, colour, and the presence of stalk
cystolith, also distribution of cystoliths in the epidermal layers of the leaf lamina have shown to be useful as
additional character for group identification in the genus Ficus.

TABLE 1. List and details of Ficus species studied.

Species Collector, Locality Date of
Collection number collection
F. annulata Blume A. Hussin, AH 16 Bukit Lagong, FRIM Kepong, Selangor 04.04.2012
A. Zainuddin, AZ 04390 Endau, Johor 10.03.1988
A. Zainuddin, AZ 6794 Kuala Berang, Terengganu 01.06.2000

F. aurantiacea Griff. B. Ummu-Hani, UHB 05 Recretional Forest of Sungai Salu, Perak 10.11.2011
B. Ummu-Hani, UHB 15 Batu Berangkai, Kampar, Perak 13.11.2011
A. Zainuddin, AZ 4665 Km 87, road to Dungun from Kuantan, 15.08.1993

F. benghalensis L. A. Hussin, AH 24 Road to Rawang, Selangor 04.04.2012

F. benjamina L. A. Hussin, AH 01 Bukit Rawang, Selangor 03.03.2012

B. Ummu-Hani, UHB 02 Section 15, Bangi, Selangor 20.10.2011
B. Ummu-Hani, UHB 30 Section 10, Bangi, Selangor 21.02.2012

F. depressa Blume B. Ummu-Hani, UHB 07 Batu Berangkai, Kampar, Perak 11.11.2011

M. Kassim, MK 315 Batu 20, Kuala Selangor, Selangor 05.10.1972

F. lepicarpa Blume B. Ummu-Hani, UHB 26 Chemerong, Recretional Forest, Terengganu 18.02.2012

Batu Berangkai, Kampar, Perak
B. Ummu-Hani, UHB 17 Pulau Tioman, Rompin 13.11.2011
A. Zainuddin, AZ 5466 28.04.1995

F. hispida L.f. A. Hussin, AH 02 Bukit Rawang, Selangor 03.04.2012

B. Ummu-Hani, UHB 48 Klinik Desa Matang, Perak 26.04.2012
B. Ummu-Hani, UHB 50 Hulu Matang, Perak 24.06.2012

F. microcarpa L.f. A. Hussin, AH 23 Road to Bukit Lagong, FRIM Kepong, 04.04.2012

Selangor
B. Ummu-Hani, UHB 25 Rintis Alsophila, Taman Paku-Pakis, FST, 21.02.2012
UKM, Selangor
B. Ummu-Hani, UHB 32 Rintis Cibotium, Taman Paku-Pakis, FST, 21.02.2012
UKM, Selangor

F. obscura Blume var. A. Hussin, AH 08 Teluk Senangin, Perak 31.03.2012

borneensis (Miq.) Corner A. Zainuddin, AZ 3865 Lenggong, Perak 29.09.1991
A. Latiff, ALM 115 Bukit Mat Deling, Taman Negara Pahang, 23.10.1982
Pahang

F. sagittata Vahl A. Hussin, AH 19 Road to Bukit Lagong, FRIM Kepong, 04.04.2012

Selangor
B. Ummu-Hani, UHB 38 Bukit Bauk, Terengganu 06.03.2012
B. Ummu-Hani, UHB 63 Kampung Sungai Rumput, Gombak, Selangor 24.12.2012

F. schwarzii Koord. A. Hussin, AH 18 Road to Bukit Lagong, FRIM, Kepong, 04.04.2012

Selangor
B. Ummu-Hani, UHB 10 Batu Berangkai, Kampar, Perak 13.11.2011
B. Ummu-Hani, UHB 28 Chemerong Recretional Forest, Terengganu 18.02.2012
F. superba (Miq.) Miq. A. Hussin, AH 10 Road to Bukit Lagong, FRIM, Kepong, 04.04.2012
Selangor
M. Kassim, MK 524 UKM, Bangi, Selangor 05.10.1989

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 A. Latiff, ALM 2658 Pulau Aur, Mersing, Johor 25.01.1988

F. tinctoria G. Forst A. Hussin, AH 15 FRIM Kepong, Selangor 02.04.2012

A. Zainuddin, AZ 5901 Pulau Tinggi, Mersing, Johor. 08.11.1995

F. ucinata (King) Becc. A. Hussin, AH 06 Kampung Bukit Rawang, Selangor 03.04.2012

F. vasculosa Wall. ex Miq. A. Hussin, AH 22 Road to Bukit Lagong, FRIM, Kepong, 04.04.2012
Selangor
A. Zainuddin, AZ 5480 Pulau Tioman, Pahang 29.04.1995
A. Zainuddin, AZ 3886 Bukit Bauk, Terengganu 20.10.1991

TABLE 2. Descriptions of cystoliths structure in Ficus species studied.

Types of Descriptions
Illustrations cystoliths
morphology
1 Cystoliths adjacent to the adaxial epidermis
Oblong-shaped
Colour: blue
Sizes: cystoliths 50 m wide and 60 -100
m long
Stalk length: 30 m long

2 Cystoliths adjacent to the adaxial epidermis

Cigar-shaped
Colour: violet
Size: cystoliths 30 m wide and 115 m
long
Stalk: absent

3 Cystoliths adjacent to the abaxial epidermis

Solitary rounded cystoliths with blunt ends
Colour: greenish
Size: cystoliths 60-70 m wide and 40-50
m long
Stalk length: 35-40 m long

4 Cystoliths adjacent to the abaxial epidermis

Bean-shaped
Colour: blue
Size: cystoliths 30 m wide and 15 m
long
Stalk: absent

5 Cystoliths adjacent to the abaxial epidermis

Y-shaped or double cystoliths
Colour: violet
Size:cystoliths 20-25 m wide and 15 m
long
Stalk length: 50 m long

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 6 Cystoliths adjacent to the abaxial epidermis
Solitary elongated cystolith with pointed ends
Colour: blue
Size: cystoliths 25-30 m wide and 35-40
m long
Stalk length: 30 m long

7 Cystoliths adjacent to the abaxial epidermis

layer
Solitary rounded-shaped
Colour: blue
Size: cystoliths 70 m wide and 75 m
long
Stalk length: 20 m long

ACKNOWLEDGMENTS
The authors wish to thank the Herbarium of Universiti Kebangsaan Malaysia, Bangi for providing dried leaves
samples. We would like to thank Mohamad Ruzi Abdul Rahman, UKMs research officer and Abu Hussin Harun
science officer of Forest Research Institute of Malaysia (FRIM), in Kepong for their technical assistance in the field.

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