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Anatomical Studies of Withania Somnifera

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Short Communication

Anatomical Studies in Withania Somnifera L. (Dunal)

- an Important Medicinal Plant
*M.Senthil Kumar1, D.Vinoth Kumar2, A.Ramachandran3 and A.Shajahan1

Abstract
Withania somnifera L. Dunal is commonly known as Aswagandha. It is one of the most important medicinal plants and also
known as Indian ginseng. The anatomical characteristics of the leaf, stem and root of W. somnifera have been studied. The
analysis provided here by studying the cross sections of root, stem, leaf, venation pattern, and trichome comprises the species.
The results showed that there is concentration of vascular bundles at the central portion of root cortex, calcium oxalate
presence in the root powder and non glandular trichome were present in the leaves.
Keywords: Anatomical features; Leaf; Stem; Root Withania somnifera; Solanaceae

Materials and Methods

and blue to the protein bodies. For studying the stomatal morphology,
venation pattern and trichome distribution paradermal sections (sections
taken parallel to the surface of leaf) as well as clearing of leaf with 5%
sodium hydroxide were prepared. Glycerin mounted temporary
preparations were made for macerated/cleared materials. Powdered
materials of different parts were cleared with NaOH and mounted in
glycerin medium after staining. Different cell component were studied
and measured. Photographs of different magnifications were taken with
Nikon Labphot 2 Microscopic unit. For normal observations, bright field
was used. For the study of crystals, starch grains and lignified cells,
polarized light was employed. Under the polarized light they appear
bright against dark background. Magnifications of the figures are
indicated by scale-bars. Descriptive terms of the anatomical features are
as given in the standard anatomy books (Cutler, 1978).

Specimen collections

Results and Discussion

The plant specimens for the Withania somnifera were collected from
Tanjore, Perambalur and Trichirapalli districts of Tamil Nadu during
June, 2009. Care was taken to select healthy plants. The required sample
of leaf, stem, and root were cut and removed from the plant and fixed in
FAA (Farmalin 5 ml + Acetic acid - 5 ml + 70% Ethyl alcohol 90 ml).
After 24 hrs of fixing, the specimens were dehydrated with graded series
of tertiary butylalcohol. Infiltration of the specimens was carried by
gradual addition of paraffin wax (melting point 580600C) until TBA
solution attained supersaturation. The specimens were cast into paraffin
blocks.

The leaf is dorsiventral with prominent midrib and bilaterally

symmetrical lamina (Fig. 1a). The midrib has broad abaxial hump and
still broader, semicircular abaxial part. The midrib consists of a wide,
arch shaped shallow bicollateral vascular bundle and heterogeneous
growing tissue. The xylems elements are narrow, thick walled and are
disposed in short radial files. The lamina is 270 m thick. Epidermal
trichomes are abundant especially on the abaxial side of the lamina. The
lateral veins are prominent and form distinct reticulate venation pattern
(Fig. 1b). The vein islets are well defined and distinct with thick
boundaries. The islets are polyhedral in outline and random in
orientation. The vein terminations are thick and comprise three or four
rows of trachieds. They may be short or long unbranched or branched.
The leaf mesophyll of Salvia forskahlei species is entirely
parenchymatic and the vein of leaf surrounded by collenchymas
(Metcalfe and Chalk, 1972). The non glandular, trichomes are abundant
on the lamina. They are densely crowded and random in distribution
(Fig. 1c). The trichomes are predominatly branched and dendroid. The
cells of the trichomes are wide rectangular in vertical plane and thin
walled. The terminal cells are pitted and the pits are not prominent. The

Introduction
Withania somnifera L. Dunal is an important medicinal plant belongs to
the family Solanaceae. It is known classically for its rejuvenating
properties, and hence called Indian Ginseng (Singh and Kumar, 1998).
It is distributed in Maharastra, Gujarat, Rajasthan, Madhya Pradesh,
Uttar Pradesh, Punjab, Himachal Pradesh, Jammu and Kashmir and
Tamil Nadu. There are ten species under this genus, of which three
species W. somnifera, W. coaguland and W. obtusifolia are found in India.
The present paper is aimed to study the anatomical features of leaf, stem,
root, venation pattern, trichome and crystal comprises of
W. somnifera and this could be utilized in the biosystematics of this plant.

Sectioning (sample preparation)

The paraffin embedded specimens were sectioned with the help of rotary
microtome. The thickness of the sections was 1012 m. The sections
were stained with Toluidine blue as per the method published by
Buscalioni (1986). Since Toluidine blue is a polychromatic stain, the
staining results were remarkably good and some cytochemical reactions
were also obtained. The dye rendered pink colour to the cellulose walls,
blue to the lignified cells, dark green to suberin, violet to the mucilage

1. Plant Molecular Lab, PG and Research Dept. of Botany, Jamal Mohamed College (Autonomous), Tiruchinapalli, Tamilnadu.
2. Department of Biotechnology, Adhiyamaan Engineering College, Hosur, Karnataka. 3. Director in Centre for Climate Change and Adaptation,
Anna University, Chennai, Tamilnadu. *Corresponding Author: senthil2323@gmail.com
43 l Advanced Biotech. Vol. 10

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Short Communication

1.Anatomical Studies of Withania Somnifera, M.Senthil Kumar et al.,

Fig.1 Sections of the leaf of the Withania somnifera studied

Fig. 1a Epidermal morphology

Fig. 1b Venation pattern

VI Vein islets, VT Vein termination,
LV Lateral Vein, MR Midrib
GT Ground Tissue, Abs Abaxial side
BVB Bicollateral vascular

ADH Adaxial hump, Col Collenchyma,

EP Epidermis, PM- Palisade mesophyll
SM Spongy mesophyll, Tr Trichome
AbE Abaxial epidermis

Fig. 1c Trichome distribution (Ep Epidermal cells; Tr Trichome)

44 l Advanced Biotech. Vol. 10

Fig 1d - Enlarged view of vascular bundle

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Short Communication

Anatomical Studies of Withania Somnifera, M.Senthil Kumar et al.,

Fig 2: Sections of the stem and root of the Withania somnifera studied

Fig. 2aAnatomy of the stem

Fig. 2b Anatomy of the root

Col Collenchyma, Ep Epidermis

Mph Medullary Phloem, Pi Pith
Oph Outer Phloem, Ph Phloem
Pc Parenchyma, Vr Vascular ray

Pe Periderm, Sph- Secondary xylem,

VC- Vacular Cylinder, OX Outer Xylem
CX Central Xylem, GP Ground Parenchyma
sectional view with rough surfaced periderm. The periderm in narrow
and not well defined. The cortical portion is wide measuring 400 m and
consists of radially compressed thin walled parenchyma cells. Inner to
the cortex is a wide and the outer vascular cylinder is produced by a
cambial zone. The middle zone in the widest part of the root is small
radial segments of xylem and small nests of phloem. The central part of
the root has a solid core of vessels and fibred, the xylem core is irregular
which are up to 50 m wide (Fig. 2ac). The presence of calcium oxalate
crystals in the root is an interesting feature in solving taxonomic
problems and used is many plant species. The Araceae family plants that
possess five types of calcium oxalate crystals (Okoli, 1988; Edoga and
Okoli 1995; Mbagmu, 2005). The analysis in this account provides the
first detailed description of anatomy of W. somnifera, from the
anatomical observations it could be concluded that the investigated
forms of W. somnifera.

References
Buscalioni, L., 1986. Studi sui cristali di ossalato de calico. Malpighia,
9:469533.
Cutler, D.F., 1978. Applied plant anatomy. Longman, London, 125128.
Fig. 2c Distribution of crystal and starch grains
(Scr Sand crystal; Pi Pith cells; PM Palisade mesophyll;
SM Spongy mesophyll)
trichomes range in height from 100 to 450 m wide. The glandular and
non glandular trichomes hairs distributed as in the leaf (Metcalfe and
Chalk, 1950). The stem is circular in sectional view with shallow ridges
and furrows. It consist a thin epidermis, fairly wide cortex, a hollow thick
vascular cylinder and wide pith. The epidermal cells are small, having
thin cuticle. The cortex consists of an outer zone of four or five layers of
collenchyma and two or three layers of inner parenchyma and the
vascular has outer and inner phloem. The xylem consist wide, thick
walled lignified fibres. The vascular cylinder including outer and inner
phloem is about 500 m thick (Fig. 1d). The stem and a well developed
collenchyma, supporting tissue at the corners of stem in Salvia species
plants (Metcalfe and Chalk, 1972). The woody stem of Salvia forskahlei
has sclerenchyma upon the phloem and also a sclerenchymatic ring upon
the sclerenchyma (Ozdemir and Senel, 2001). The root is circular in

Edoga, H.O., and Okoli, B.E., 1995. Histochemical studies in the leaves
of some Dioscorea L. and the taxonomic importance. Feddes Report,
106:113120.
Mbagmu, 2005. Anatomical studies on the root of some Vigna savi
species Agricultural journal 1(1):810. Medwell Online. Pakistan.
Metcalfe, C.R., and Chalk, L., 1950. Labiatae, In: Anatomy of the
Dicotyledons. Oxford Clarendon Press, London, 2:1041-1053
Metcalfe, C.R., and Chalk, L., 1972. Anatomy of the Dicotyledons,
Clarendon Press, Oxford 2:1041-1053.
Okoli, B.E., 1988. On the probable function and taxonomic value of
calcium oxalate crystals in cucurbitaceae. Feddes Repert, 99:139-142.
Ozdemir, C. and Senel, 2001. The morphological anatomical and
karyological properties of Salvia forskahlei L. (Lamiaceae) in Turkey.
J Econ Tax Bot 19: 297-313.
Singh, S., Kumar S., 1998. Withania somnifera The Indian Ginseng
Aswagandha. Central Institute of Medicinal and Aromatic Plants.

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