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Int. J. Life. Sci. Scienti. Res.

eISSN: 2455-1716
Das, 2018
DOI:10.21276/ijlssr.2018.4.6.8

Research Article

Karyomorphological Studies in Three Species of Alocasia (Schott.)


G.Don.- An Ethno-medicinally and Economically Important Genus
*
Bandana Nabis Das

Associate Professor, Department of Botany, Handique Girls’ College, Guwahati, Assam, India

*Address for Correspondence Dr. Bandana Nabis Das, Associate Professor, Department of Botany, Handique Girls’
College, Guwahati, Assam, India

Received: 08 Apr 2018/ Revised: 05 Jul 2018/ Accepted: 29 Oct 2018

ABSTRACT
Karyomorphological studies in Alocasia macrorrhiza (L.) G.Don., Alocasia fornicate (Roxb.) Schott, Alocasia longiloba Miq.
belonging to the family Araceae using root tip squash technique was carried out. It was observed that the chromosome number
for the three species was found to be 2n=28 and chromosomes are smaller in size. The chromosomes in Alocasia longiloba were
found to be longer in length in comparison to Alocasia macrorrhiza, Alocasia fornicata. Present studies also reveal that the
karyotype is a symmetric type. The present karyomorphological study has been undertaken as it is an established fact that
karyomorphological analysis forms a prerequisite for the genetic improvement of any plant species. This study would be helpful in
the protection, conservations of the species by establishment of germplasm bank.

Key-words Alocasia macrorrhiza, Karyomorphology, Karyotypes, Symmetric type, Chromosomes

INTRODUCTION
The genus Alocasia (Schott) G. Don belongs to the family Boiled stems of A. macrorrhizos are used as a laxative,
Araceae consisting of about 2,500 species [1]. There are chopped-up roots and leaves as a rubefacient, and juice
79 species native to tropical and sub-tropical Asia to from the petiole against a cough. The plants are applied
Eastern Australia and wildly cultivated elsewhere [2]. The for stimulating the skin, e.g. in cases of fever and to
species of this genus have been distributed in the remove blotches. The rhizome is sometimes used as a
Northeast and South India, Sri Lanka, and Bangladesh [3]. poultice to treat furuncles. The pounded stems are
These are generally grown in marshy lands, but applied as a paste to snakebites and scorpion stings.
sometimes also in shady damp places in the forest and in Alocasia fornicata (Roxb.) Schott is important as food
village thickets. In this study, three species of Alocasia and ethno-medicine in Asia and Africa since time
(Schott) G.Don, A. macrorrhiza (L.) G.Don, A. fornicate immemorial. In India, many people have been using
(Roxb.) Schott, A. longiloba Miq. were selected. These rhizome paste to treat wounds, cure heel cracks and kill
species are ethno-medicinally and economically worms in domestic animals. A number of chemicals could
important plants. According to IUCN Red list Alocasia be isolated from this species. The antioxidant properties
atropurpurea and Alocasia sanderiana are tagged as of the edible parts of different plants have already been
Critically Endangered [4]. Several medicinal applications established [5]. There is growing industrial interest
of Alocasia have been reported for South-East Asia. towards the production of herbal antioxidants that can
be supplemented in medicine, cosmetics, and
How to cite this article
Das BN. Karyomorphological Studies in Three Species of Alocasia nutraceuticals in the modern world.
(Schott.) G.Don.-An Ethno-medicinally and Economically Important The tissues of Alocasia contain calcium oxalate crystals,
Genus. Int. J. Life. Sci. Scienti. Res., 2018; 4(6): 2116-2121.
which produce irritation of the skin and inflammations of
the oral cavity and mucous membranes. Sapotoxin is also
Access this article online
present, and the toxic effects include gastroenteritis and
www.ijlssr.com paralysis of the nerve centres. Hydrocyanic acid is often
present. The poisonous substances can be removed by
repeated cooking, but the rhizomes and bases of petioles

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Int. J. Life. Sci. Scienti. Res. eISSN: 2455-1716
Das, 2018
DOI:10.21276/ijlssr.2018.4.6.8

of A. macrorrhizos which are sometimes used for food For the preparation of slides the root tips were first
usually contain few poisonous substances. A lectin has hydrolysed with 1N HCl and stained with aceto orecin
been isolated from the rhizome of A. macrorrhizos, and warmed over flame for 10 to 15 minutes and kept
which showed potent mitogenic activity on human for 2 - 3 hours at room temperature. Single root tip was
peripheral blood lymphocytes in the [3H]-thymidine taken in a drop of 45% acetic acid on a slide. Only the
uptake assay. It was a T-cell mitogen and did not induce dividing tip region was taken discarding the other tissue.
any appreciable DNA synthesis in B-enriched Cover slip was placed over the tip and squashed by
lymphocytes. This species also contains a protein which applying uniform pressure with the thumb through a
inhibits both the enzymes trypsin and chymotrypsin. The piece of blotting paper, a gentle tapping followed by
seed extract showed antifungal activity. There is also a heat fixing and finally sealed with paraffin for further
report on HIV-1 protease inhibitory activity [6]. studies [8,9].
Alocasia is a plant of great economic value. The temporary slides thus prepared were observed
Experimentally antimicrobial, antifungal, antioxidant, under compound microscope at a magnification of 1000x
hepato-protective, anti-diarrheal, anti-protozoal, using oil immersion (10×100x, oil immersion). This
anticancer properties have been found in many species procedure was standardized through trial and error
of Alocasia. As such it needs conservation and detailed method. Metaphase plates were selected for
chromosome studies would aid in creating germplasm [7]. karyomorphological analysis of the chromosomes.
Perfectly stained chromosomes were photographed
Hence, the present study has been undertaken: using Trinocular microscope-N400-M, CMOS camera 5M
To conduct karyomorphological studies in three species with image analysis system. The drawings of the
of Alocasia (Schott) G.Don: chromosomes were made with the help of camera lucida
o Alocasia macrorrhizos (L.) G.Don apparatus. Idiograms were then constructed on tracing
o Alocasia fornicate (Roxb.) Schott paper.
o Alocasia longiloba Miq.
Following parameters of the chromosomes were
MATERIALS AND METHODS
considered:
In the present investigation three species of Alocasia
i) Length of the long arm
(Schott) G.Don were collected from different localities of
ii) Length of the short arm
Greater Guwahati. The saplings were potted inside the
iii) Total length of the chromosome
institute campus for collection of fresh roots for the
iv) Volume of the chromosome
experiment. The following investigation proceeded for 5
v) Relative length
months i.e. from January to May 2017.
vi) Centromeric position
Cytological investigation- For chromosome
On the basis of the length, the chromosomes lengths
characterization in A. macrorrhiza, A. fornicata and A.
were categorized under the following types-
longiloba detailed karyotypic studies were undertaken.
Karyotypes were prepared from the somatic Type A= 3.00 µm and above
chromosomes. For cytological studies, root tips squash Type B= 2.50 µm - 2.90 µm
technique is done. Very young root tips were collected Type C= 2.00 µm - 2.49 µm
from the plant between 7:45 AM - 8:15 AM washed in Type D= 1.00 µm - 1.90 µm
double distilled water and pre-treated with saturated Type E= 0.01 µm - 0.99 µm
solution of Para dichlorobenzene at a suitable
temperature for 3 hours at 4°C±2°C. Pretreated young The volume of an individual chromosome was calculated
root tips are fixed in a suitable fixative such as Carnoy’s as chromosome volume-
fluid-2 (1:3:6; Glacial acetic acid: Chloroform: Ethanol) (V) = πr2h
for 24 to 28 hours at room temperature. After fixation
Where, r= Radius of the chromosome; h= Length of the
the root tips were thoroughly washed with 70% ethanol
whole chromosome.
and finally they were stored used for cytological work.

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The total chromosome volume was then expressed by classified into metacentric, sub-metacentric, sub-
adding the volumes of all the chromosomes. On the basis telocentric, and telocentric following the nomenclature
of centromere position, the chromosomes were system of Levan et al. [10].

Fig. 1 (a-d): Karyotype and Ideogram of somatic chromosomes of Alocasia macrorrhiza (L) G.Don

Fig 2 (a-d): Karyotype and Ideogram of somatic chromosome of Alocasia fornicate (Roxb.) Schott.

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Int. J. Life. Sci. Scienti. Res. eISSN: 2455-1716
Das, 2018
DOI:10.21276/ijlssr.2018.4.6.8

Fig 3 (a-d): Karyotype and Idiogram of somatic chromosomes of Alocasia longiloba Miq.

RESULTS
Cytological and karyotypic investigation in A. macrorrhiza chromosomes belongs to type B. So the Chromosome
(L) G.Don, A. fornicata (Roxb.) Schott. and A. longiloba formula can be written as: A15+B13+C0+D0+E0= 2n= 28,
showed that all the three species consist of 2n=28 is the and the karyotypic formula can be written as: M10+
chromosome (Table 1). On the basis of length of Sm18= 28. However, A. longiloba Miq. 20 chromosomes
chromosomes, it was observed that in A. macrorrhiza 10 belongs to type A and 8 chromosomes belongs to type B,
chromosomes belongs to type A and 18 chromosomes therefore their chromosomes formula can be written as:
belongs to type B and the chromosomes formula can be A20+B8+ C0+ D0+E0= 2n= 28 and the karyotypic formula
written as: A10+ B18+ C0+D0+ E0= 2n= 28. Depending on can be written as: M8+ Sm20= 28. The microphotograph,
the position of their centromere, the karyotypic formula camera lucida diagram, karyotype and idiogram of the
can be written as M17+Sm11= 28. In A. fornicate (Roxb.) chromosomes are represented in Fig. 1 A, B, C, and D
Schott. 15 chromosomes belongs to type A and 13 respectively.

Table: 1: Details of karyotype analysis in three species of Genus Alocasia (Schott) G. Don

Range of Chromosomes
Chromosome No.

Types of Karyotype
Arm ration
(2n)

Taxa
Relative

volume

chromosomes formula
length

length

radius

Alocasia
1.5 µm to
macrorrhizos (L.) 28 2.7-4.9 0.50 1.5-3 1-1.5 A10+ B18 M17+Sm11= 28
3.5 µm
G.Don
Alocasia
1.5 µm to
fornicate (Roxb.) 28 2.4-4.8 0.50 1.5- 3.2 1-1.5 A15+ B13 M10+Sm18= 28
3.5 µm
Schott.

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Int. J. Life. Sci. Scienti. Res. eISSN: 2455-1716
Das, 2018
DOI:10.21276/ijlssr.2018.4.6.8

Alocasia 1.5 µm to
28 1.9- 4.9 0.50 1.5-4 1-1.6 A20+ B8 M8+Sm20= 28
longiloba Miq. 3.5 µm

DISCUSSION posses 2n = 28 chromosomes. A centromeric formula of


It is well known that karyotype analysis often plays an M17+ Sm11= 28 was found in A. macrorrhiza (L,) G.Don,
important role in determining the taxonomic status of a M10 +Sm18= 28 in A. fornicata (Roxb.) Schott. and M8
taxon where the taxonomic parameters are insufficient, +M20=28 in A. longiloba Miq. A. macrorrhiza (L,) G.Don,
because the karyotype indicates a very stable character showed more symmetric karyotype consisting of
that is specific for each specimen. However, a problem maximum metacentric chromosomes. The other two
arises when different taxa possess the same forms have relatively fewer metacentric chromosomes
chromosome number and similar karyotype features. In and more sub-metacentric chromosomes. This result
this situation, it is hard to distinguish between different shows the karyotypic relatedness between the species,
taxa by conventional karyotype analysis. Even the which could be taken as a positive output to do further
consideration of chromosome length, arm ratio, position, works on the commonly available species of this genus,
and number of secondary constrictions are not always so as to conserve the other threatened species of this
sufficient to differentiate individual chromosomes. same genus. The following study also reveals the
Minute alterations regarding the distribution pattern of symmetric karyomorphology between species which
GC- and AT-rich repeats in the karyotypes cannot be shows the primitiveness in this genus [15,16].
detected through conventional karyotype analysis.
CONCLUSIONS
Moreover, the deletion of heterochromatic regions may
There is growing industrial interest towards the
change the karyotype of a specimen without affecting
production of herbal antioxidants and aim to evaluate
the morphology [11].
the biological activities, pharmacological applications
Different scientists have studied karyotype and RAPD
and clinical studies of Alocasia macrorrhiza (L.) G.Don,
Analysis of Three Morphological Forms of Alocasia
in an attempt to provide a direction for further research.
fornicata (Roxb.) Schott. The three forms of Alocasia
It is an established fact that karyomorphological analysis
fornicata (Roxb.) Schott were found to possess 2n=28
forms a prerequisite for the genetic improvement of any
chromosomes. Similar diploid chromosome numbers
plant species. Therefore the detailed cytological
were reported [12,13].
investigation has been undertaken in these species of
Singh et al. [14] in their article, aim to evaluate the
Alocasia (Schott.) G.Don. Realizing their importance and
biological activities, pharmacological applications and
utility, ex-situ conservation measures may be initiated
clinical studies of Alocasia macrorrhiza in an attempt to
for protection, preservation, and regeneration of such
provide a direction for further research. They found that
medicinally and economically important plants.
apart from household decorative purposes, this plant has
some pharmacological activity like antifungal, weak CONTRIBUTION OF AUTHORS
hemagglutinating activity, antidiuretic, laxative, All authors equally contributed in this article.
antitubercular and reduces the activity of human
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Int. J. Life. Sci. Scienti. Res. eISSN: 2455-1716
Das, 2018
DOI:10.21276/ijlssr.2018.4.6.8

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