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Abstract

Medicinal plants Tephrosia villosaand Cassia tora (Linn.) are belongs to the family Fabaceae and widely distributed in India and other tropical regions. Different parts (roots, stem, leaves and seeds) of the both plants have been used in various traditional medicare systems for ailments of human being. In the present study deals with the evaluation of pharmacognostic parameters in terms of the macro and micro-morphological characters and quantitative profiles of the leaves of Tephrosia villosa andCassia tora(Linn.). The observations of the current study can be useful for identification, standardization and authentication of plants.

Introduction

The nature has provided us depositories of medication to cure all ailments of humans. The traditional herbal medicines are still utilized in rural and tribal area of our country. The traditional herbal medicines are still utilized by a large part of community and conventional practitioners, who depends on natural medication for their primary healthcare system 1. According to the World Health Organization, the macroscopical and microscopical description of a medicinal plant is the first step towards establishing the identity and degree of their purity 2. Tephrosia villosais commonly known as Ruvali-biyani and Kharanio. This plant is an annual, gregarious, erect bushy herb, up to 9 cm high which is widely distributed in Southern Asia, and in India throughout in plain 3. It is very useful plant due to the presence of various phytochemicals. Aqueous extract of T. villosa leaves is used as herbal remedy in the treatment of diabetes mellitus in India. In Africa, the herb is used as green manure to improve the soil4. Cassia tora Linn. is commonly known as Foetid Cassia, Tora, Sickle Senna, Wild Senna, Sickle Pod, Coffee Pod, Tovara, Chakvad and Ringworm Plant. It is also known as Charota and Chakvad in Hindi, Chakramarda & Dadmari in Sanskrit5. It is an annual fetid herbaceous plant which is widely distributed in India, Ceylon, China and other tropical countries. Different parts of plant Cassia toraare widely used to cure various diseases like skin diseases, eye diseases, liver diseases etc.6. The present studies were carried out to determine the macro and micro-morphological characters, and quantitative parameters to evaluate pharmacognostic properties of the leaves of Tephrosia villosaand Cassia tora Linn.

Materials And Methods

Material and Methods Collection and Authentication of Botanical Material

Fresh leaves of Tephrosia villosa andCassia tora (Linn.)were collected from the local area around Ajmer, Rajasthan (India). A herbarium specimen of each plant has been deposited at the Department of Botany, Samrat Prithviraj Chauhan Government College Ajmer.

Preparation for Examination

Collected fresh leaves of plantsTephrosia villosa andCassia tora (Linn.) were washed with water and cleared of foreign matter and other contaminants. Microscopic evaluation was done by using standard methods.

Chemicals

The chemicals like lactic acid, trichloroacetic acid, phenol, safranin, glycerine, distilled water etc. were used. The entire chemical, used in experiments were of analytical grade.

Macroscopic Examination

The fresh leaves of both plants were evaluated for various organoleptic parameters such as the shape, size, colour, margin, texture, apex, presence or absence of petiole, phyllotaxy etc.

Microscopic Examination

The histological features such as structure of epidermal cells, structure, distribution and type of stomata, structure and distribution of trichomes on the fresh leaves have been analyzed.

Quantitative Microscopy

For the quantitative microscopy the number of epidermal cell, number of stomata and stomatal index for both abaxial and adaxial surfaces of the leaves were determined according to method of Mohan Ram and Nayyar7 with slight modifications. Entire fresh leaves were soaked in the mixture of trichloroacetic acid and phenol in the proportion of 2:1 for 15-60 min at 60˚C temperature. The cleared leaves were flooded with pure lactic acid and kept at 60˚C temperature for various time periods and were then washed with water. Cleared leaves were stained with safranin, mounted with glycerin and observed under compound microscope. The quantitative estimations were carried out by using methods of Salisbury (1927); 8; 9; 10

Epidermal cell number: The average number of epidermal cells per square millimeter unit of epidermis is termed as epidermal cell number.

Stomatal number: The average number of stomata per square millimeter of epidermis is termed as stomatal number.

Stomatal index: The percentage of number of stomata to the total number of epidermal cells (wherein each stomata also is considered to be a single epidermal cell), is termed as stomatal index. The stomatal Index was calculated using the standard formula given below.

Stomatal Index (S.I.) =S*100/S+E

S = quantity of stomata per unit area and E = number of ordinary epidermal cells in the same unit area 1112.

The quantitative parameters have been subjected to statistical analysis and expressed as Mean ± SEM.

Results

Results and Discussion Macromorphology

The results of the macroscopic and organoleptic examination established that the plant Tephrosia villosa is an annual gregarious, erect bushy herb. Stem and branches angular, densely hairy with white appressed hairs. Leaves cauline and ramal, odor less, pinnately compound (imparipinnate), stipulate (free lateral). Leaflets alternate/opposite/ superposed, sessile, entire margin with dense hairs, oblanceolate in shape, apex emarginate, 10-14, pale green, more or less glabrous on upper surface and densely hairy on lower surface, having reticulate unicostate venation. Trichomes (hairs) are unicellular with glandular base. Figure 1.

Figure 1. a. Plant of Tephrosia villosa b. Twig of Tephrosia villosa c. Leaves of Tephrosia villosa showing adaxial and abaxial surfaces

The results of the examination reveals that the plant Cassia tora (Linn.) an annual herbaceous malodorous herb or under shrub. Stem is cylindrical, branched and grows in rainy season. Leaves cauline and ramal, pinnately compound (paripinnate), stipulate (free lateral). Leaflets are alternate/ Opposite/Superposed, numbers are 6, Obovate, entire margin with dense hairs, apex obtuse, dark green on adaxial surface and light green on abaxial surface, glabrous on adaxial surface and some trichomes are present on abaxial surface, conical shaped gland present at the base of lower leaflet. Terminal leaflet are larger than lower leaflet with dimension 10.29±0.95 and 4.07 ±0.23 respectively and having reticulate unicostate venation. Trichomes are unicellular. Figure 2, Table 1.

Figure 2. a. Cassia tora plant b. leaf of Cassia tora c. leaflets showing adaxial and abaxial surfaces d. conical gland present on base of lower leaflets

Table 1. Organoleptic Parameters.

S.No.	Parameters	Tephrosia villosa	Cassia tora (Linn.)
1.	Bearing of leaf	Cauline and Ramal	Cauline and Ramal
2.	Odour	Obnoxious/ displeasing	Obnoxious/ displeasing
3.	Leaf type	Pinnately Compound / Imparipinnate	Pinnately Compound / Paripinnate
4.	Glands at leaf base	Absent	Conical shaped gland present at the base of lower leaflet
5.	Phyllotaxy of leaflets	Alternate/Opposite/Superposed	Alternate/Opposite/Superposed
6.	Outgrowths at base of leaf	Stipulate/ Free lateral	Stipulate/ Free lateral
7.	Colour of leaf-let (adaxial surface)	Pale green	Dark green
8.	Colour of leaf-let (abaxial surface)	Pale green	Light green
9.	Petiole (of each leaf-let)	Absent/sessile	Petiolate (very short)
10.	Number of leaflet	11-17	6
11.	Composition of lamina (of each leaf-let)
a.	Shape	Oblanceolate	Obovate
b.	Dimension (l*b)	1.80±0.07	10.29±0.95/4.07 ±0.23
c.	Margin	Entire with dense hairs	Entire with dense hairs
d.	Apex	Emarginate	Obtuse
e.	Surface appearance	Glabrous on upper surfaces and Dense trichomes are present on lower surface of leaflets	Glabrous on upper surface and Small amount of trichomes are present on lower surface of leaflets
f.	Trichomes	Unicellular with glandular base	Unicellular
g.	Venation	Reticulate/Unicostate	Reticulate/Unicostate

Micromorphology

The results of the micromorphology reveal that the leaves of Tephrosia villosaconsists of amphistomatic type of stomata (stomata found equally upon the both surfaces). The epidermal peeling of both adaxial and abaxial surfaces of leaves consists of small polygonal, thin walled epidermal cells and paracytic stomata which were surrounded by 2 or 3 subsidiary cells on either side of the guard cells and were parallel to stomata. Trichomes were absent on adaxial surfaces. Dense, white, silky and unicellular trichomes with glandular base were present on adaxial surface and leaf margin. Figure 3.

Figure 3. Micromorphology of leaves of Tephrosia villosa (abaxial surface and adaxial surface)

The results of the micromorphology of the leaves of Cassia toraare that stomata were present more on upper surface and less on its lower surface. The epidermal peeling of both adaxial and abaxial surfaces of leaves consists of small polygonal, thin walled epidermal cells and paracytic stomata which were surrounded by 2 or 3 subsidiary cells on either side of the guard cells and were parallel to stomata. Trichomes were absent on upper surface. Dense, unicellular trichomes were present on entire margin where few trichomes present on lower surface. Figure 4.

Figure 4. Micromorphology of leaves of Cassia tora (adaxial surface and abaxial surface) Quantitative Microscopy

The quantitative analysis shows that the number of epidermal cells is 78.5±0.57and 69.7±0.83, stomata number is 13.9±0.5 and 15.5±0.36, stomata index is 15.02±0.50 and 18.19±0.4for adaxial and abaxial surfaces respectively for the leaves of Tephrosia villosa.

The results of quantitative analysis of leaves ofCassia tora (Linn.) reveals the number of epidermal cells is 51.4±0.7and 50.7±0.5, stomata number is 12.1±0.5and 10.1±0.37, stomata index is 19±0.49and 16.59±0.53for adaxial and abaxial surfaces respectively. Table 2, Figure 5.

Figure 5. On X-axis- parameters and on Y- axis mean value of observed data

Table 2. Quantitative Parameters

S.No.	Parameters	Tephrosia villosa	Cassia tora (Linn.)
1	Stomata number on adaxial surface	13.9±0.5	12.1±0.5
2	Stomata number on abaxial surface	15.5±0.36	10.1±0.37
3	Epidermal cell number on adaxial surface	78.5±0.57	51.4±0.7
4	Epidermal cell number on abaxial surface	69.7±0.83	50.7±o.5
5	Stomatal index on adaxial surface	15.02±0.50	19±0.49
6	Stomatal index on abaxial surface	18.19±0.4	16.59±0.53

Conclusion

The pharmacognostic profile in terms of macro and micro-morphological characters of the leaves of Tephrosia villosaand Cassia tora (Linn.) can be useful in identification, standardization and authentication of raw material. The various parameter studied during the present analysis will also be helpful in quality assessment and detection of adulterants in the herbal material used by various pharmaceutical companies.

Journal of Advances in Plant Biology