Received by Lynn Received on 2012-3-26
ID No. B175 Revised on
Pharmacognostic and pharmacological evaluation of Ruellia tuberosa L.
Rehmanullah, Muhammad Ibrar and Ishfaq Hameed
Department of Botany, University of Peshawar, Pakistan
Ruellia tuberosa Linn. was studied to investigate the macro – and
microscopical, vein islet and vein termination numbers, palisade ratio, stomatal index and different chemical parameters. The antibacterial, antifungal and phytotoxic activities of the crude extract of the plant were also determined. Five bacterial species were used. Among them Salmonella typhi, Escherichia coli, and Pseudomonas
aeruginosa were the most susceptible bacterial species having MICs 10, 4.0, and 14 mg/ml, respectively. Among the tested fungal species Fusarium solani, and
Aspergillus niger were more susceptible to crude extracts with MICs 1.34, 2.78, and 1.45 µg/ml, respectively. At the concentration of 10µg/ml the methnolic extract exhibited significant activity, at 100µg/ml the activity was good and at 1000 µg/ml the activity was moderate against Lemna minor. The above selected plants were
shown by in vitro assays to be a potential source for natural antifungal, antibacterial and phytotoxic agents.
Keywords: Ruellia tuberose, Macro-microscopical study, chemical tests, antibacterial, antifungal, phytotoxic activities.
Morphology and histology makes the first step to get knowledge about the diagnostic features, which are ascertained through the study of the tissue and their arrangement, cell wall and cell content (Youngken, 1950). Plant anatomy has been a
source of fascination and a field of scientific inquirysince the time of the earliest
microscopists. The subject matterof plant anatomy centers on aspects of structure that
can beobserved not only with the light microscope but also using transmission and
scanningelectron microscopy and other tools of cell biology (Dengler, 2002). Not all the chemical compounds elaborated by plants are of equal interest to the pharmacognosist. Until relatively the so-called "active" principles were frequently alkaloids or specific glycosides usually with pronounced pharmacological properties; these received special attention, and in large measure constituted the principle plant drugs of the allopathic system of medicine. Other groups such as carbohydrates, fats, and proteins are of dietetic importance, and many such as starches and gums are used
in pharmacy but lacked any marked pharmacological actions (Evans, 2002). Putiyanan et al. (2009) reported that the macroscopic characters were studied for sample collecting and microscopic characters of transverse section of Fak khaao’s
leaves were compared to the leaf powders showing the upper and lower epidermis, trichome, collenchyma, palisade mesophyll, spongy mesophyll, stoma (guard cell), vascular bundles, etc., which were similar to microscopic description of drug powders. The values of stomatal index, veinlet termination number, vein-islet number and palisade ratio were calculated for standardization of samples which were 11.84?1.77, 5.95?1.31, 2.38?0.40 and 4.49?0.73, respectively. Latha and Kannabiran (2006) reported the presence of tannins, saponins, flavanoides, phenolic compounds, cardiac glycosides and carbohydrates in S. trilobatum. Ruellia tuberosa is used in folk
medicine due to its diuretic, diabetic, antipyretic, analgesic, and antihypertensive properties (Chiu & Chang, 1995). Recently, it is also being used as one of the components in a herbal drink in Taiwan (Chen et al. 2006). As no such literature was
available on this plant therefore the present study was carried out. Materials and Methods
The fresh specimens of the plants were collected from the Department of Botany, University of Peshawar, Pakistan. The plants were identified with the help of Flora of Pakistan by Prof. Dr. Abdur Rashid Plant Taxonomist (Ali and Qaiser, 2007). The specimen was cleaned and washed and dried in air for 15 days and was used for different tests i.e microchemical tests. These entire specimens were ground with the help of electric grinder and were mesh to 60 and were preserved in airtight bottles. Some fresh specimens were used to study morphological characters, while some of it was utilized for different anatomical parameters like vein islets numbers, vein termination number, palisade ratio, general anatomy of the root, stem and leaf and stomatal study. The macroscopical features of the plant were determined by following Wallis, (2004). The anatomy of the root and stem was determined by following a standard method of (Puruis et al. 1966). For leaf anatomy Subrahmanyam, (1996)
method was used. The vein islet numbers, vein termination number, palisade ratio was determined by following Evans, (2002). Different chemical parameters like alkaloids, mucilage, anthraquinon derivatives, calcium oxalate, tannin, lignin, starch, fats & oil, cutin, cellulose were determined by following Evans, (2002) and protein was determined by following Johnson, (1940).
Preparation of Extracts
Whole plants of Ruellia tuberosa L. was ground to 60 mesh powder using an
electric grinder. Fifty grams of each sample was separately soaked in 250 ml 70 % methanol for 72 hours. Thereafter each plant extract was passed through Whatman filter paper No. 1823. This process was repeated for 3 times. Evaporating in a rotatory
oevaporator at 40 C was carried out to concentrate the extracts. These extract was
ostored at 4 C priors to use. The methanolic extract and the standard drug were dissolved in dimethylsulphoxide (DMSO) at the concentration of 2 mg/ml and 1 mg/ml for antibacterial, 24 mg/ml and 1 mg/ml for antifungal, 30 mg/ml and 1 mg/ml for phytotoxic activities, respectively.
The tested bacteria, fungi and insects
Both gram negative and gram positive bacterial species Escherichia coli,
Salmonella typhi, Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsella
pneumoni have been used. Nutrient agar medium was used for the growth of bacteria for agar diffusion method (Mariam et al., 1993) and nutrient broth medium for serial
dilution method (Spooner and Sykes, 1972). Fungal species Fusarium solani,
Aspergillus flavus, Aspergillus niger, Candida albicans and Candida glabarata.
Sabouraud dextrose agar (SDA) was used for the growth of fungi for agar dilution method following Mariam et al. (1993).
Antibacterial, antifungal and phytotoxic activities of the plant were determined by following a method of Ahmad et al. (2009).
Table No.1. Macroscopical features of the different parts of Ruellia tuberosa L.
sPlant Parameters Fresh Dry
Root Colour Brownish Brownish
Odour Characteristic Indistinct
Shape Cylindrical Cylindrical
Rootlets Present Present
Direction of growth Vertical Vertical
Fracture Fibrous Uneven
Texture Smooth Smooth
Stem Colour Dark Green Light Green
Odour Indistinct Indistinct
Shape Angular Angular
Phyllotaxis Opposite Opposite
Kind Herbaceous Herbaceous
Direction of growth Upright Upright
Fracture Uneven Fibrous
Texture Hairy Hairy
Leaf Colour Upper surface dark green Both surfaces light
lower surface light green green
Dimension Length is cm 2.3 cm, width is Length is cm 2.2 cm,
cm 1.1cm width is cm 1.0 cm
Incision Nil Nil
Composition Simple Simple
Venation Reticulate unicostate Reticulate unicostate
Margin Wavy Wavy
Apex Obtuse Obtuse
Surface Pubescent Pubescent
Shape of leaf Cordate Cordate
Table 2. Anatomical features of the different parts of the Ruellia tuberosa L.
Plant cell Value Root Stem
L(µm) W(µm) L(µm) W(µm)
22 10 34 27 Epidermis Minimum
38 19 55 39 Maximum
26 12 39 31 Mean
12 9 18 13 Xylem Minimum