Powder Microscopic analysis on Leaf, Petiole, Stem and Root on Thespesia populnea (L.) Soland ex Correa

 

S. Muthukumar, N. Sami Veerappa*

Department of Botany, Periyar EVR College (Autonomous), Tiruchirappalii – 620 023, Tamil Nadu, India.

*Corresponding Author E-mail: prof.nsveerappa@gmail.com

 

ABSTRACT:

Thespesia populnea is a reputed ever green tree belonging to the family Malvaceae; commonly known as Indian tulip tree. The plant is distributed tropical regions and coastal forest in India. It is well known and all the parts are used in traditional system of medicine. The plant has been used as astringent, antibacterial, hepatoprotective, haemostatic, anti-diarrhoeal and anti-inflammatory. The scientific parameter is necessary to identify the exact plant material and to find its quality and purity. The present study deals with powder microscopically studies plant parts like leaf, petiole, stem and root. These studies indicated the possible information for correct identification and standardization of this plant material.

 

KEYWORDS: Thespesia populnea, leaf, petiole, stem and root, powder microscopy.

 

 

 

INTRODUCTION:

Thespesia populnea Soland ex Correa (family Malvaceae) is medium sized evergreen tree averaging 6 -10 m (20 - 33 ft) in height, with a short, often crooked stem and a broad, dense crown. It has glossy green, heart-shaped leaves and yellow hibiscus-type flowers. The tree grows well along warm coastal areas from the east coast of Africa and South and Southeast Asia to Melanesia, Micronesia, and Polynesia. It is currently naturalized in tropical climates throughout the world from the Caribbean to Africa. The tree is valuable as a coastal windbreak because it is highly resistant to wind and salt spray and grows well in sandy, saline soils 1. It propagates easily and grows rapidly. It naturalizes easily and has become a weed in some areas, so it should not be planted in areas where it is not already present.

 

The tree grows best under full sunlight and tolerates drought conditions. The heartwood is resistant to dry wood termites. Milo has many uses including coastal protection, animal fodder, windbreaks, and living fences. The most common use in the Pacific today is probably as an ornamental tree, despite its valuable timber 2.

 

Thespesia populnea is a large tree found in the tropical regions and coastal forests in India and cultivated in the gardens. All the parts of the plant used in traditional system of medicine. The bark, leaves, flower and fruits are useful in cutaneous infection such as scabies, psoriasis, eczema, ringworm, and guinea worm. The decoction of the bark is commonly used for the treatment of skin and liver diseases. The barks and flowers posses astringent, hepatoprotective, antioxidant and anti-inflammatory activities in rats3, 6.The leaves and bark of this tree are still used to produce oil for the treatment of fracture wounds and as an anti-inflammatory poultice applied to ulcers and boils, as a folk medicine6. Gossypol was found to be the major component of Thespesia populnea7 producing anti-fertility effects in rats8,9 as well as in human beings10. Four naturally occurring quinones viz thespone, thespesone, mansonone-D, and mansonone-H have been extracted from heart wood of the plant11. The phytochemical study of bark reveals the presence of gossypol, tannin and coloring matter12. Lack of proper standard of medicinal plants may lead to usage of substandard drugs which will cause damage to the faith on traditional system of medicine. Therefore scientific method must be developed to identify and maintain quality of plant drugs. With this aim the present investigation was planned to study the powder microscopic aspects of Thespesia populnea whole plant.

 

MATERIALS AND METHODS

Collection of specimens:

The plant specimens for the proposed study were collected from Periyar EVR College campus, Tiruchirappalli, Tamil Nadu, India. Care was taken to select healthy plants and normal organs. The required samples of different organs were cut and removed from the plant and fixed in FAA (Farmalin-5ml+ Acetic acid-5ml + 70% Ethyl alcohol-90ml). After 24 hrs of fixing, the specimens were dehydrated with graded series of tertiary -Butyl alcohol as per the schedule13. Infiltration of the specimens was carried by gradual addition of paraffin wax (melting point 58- 60oC) until TBA solution attained super saturation. The specimens were cast into paraffin blocks.

 

Sectioning:

The paraffin embedded specimens were sectioned with the help of Rotary Microtome. The thickness of the sections was 10-12 μm. Dewaxing of the sections was by customary procedure14. The sections were stained with Toluidine blue as per the method15. 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, blue to the protein bodies etc. wherever necessary sections were also stained with safranin and Fast-green and IKI(for Starch).

 

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 or epidermal peeling by partial maceration employing Jeffrey’s maceration fluid13 were prepared. Glycerine mounted temporary preparations were made for macerated/cleared materials. Powdered materials of different parts were cleared with Naoh and mounted in glycerine medium after staining. Different cell component were studied and measured.

 

Photomicrographs:

Microscopic descriptions of tissues are supplemented with micrographs wherever necessary. Photographs of different magnifications were taken with Nikon labphoto 2 microscopic Unit. For normal observations bright field was used. For the study of crystals, starch grains and lignified cells, polarized light was employed. Since these structures have birefringent property, under polarized light they appear bright against dark background. Magnifications of the figures are indicated by the scale - bars. Descriptive terms of the anatomical features are as given in the standard Anatomy books16.

 

 

FIG-1: Thespesia populnea - Fragments adaxial epidermal as seen in surface view

ACW: Anticlinal walls; CoC: Costal Cells;

EC: Epidermal Cells

 

 

FIG-2: Adaxial epidermis showing cuticular ridges 

ACW: Anticlinal walls; CuS: Cuticular Striatious        

 

 

FIG-3: Abaxial epidermis showing Stomata 

EC: Epidermal Cells; SC: Subsidiary Cells; St: Stoma  

 

 

FIG - 4: Thespesia populnea - Calcium Oxalate crystals as seen under polarised light            Cr: Crystals

 

 

 

FIG- 5 and 6: Thespesia populnea - Peltate epidermal scales

PlS: Peltate Scales RaS: Radiating Scales; StC: Stalk Cells       

 

 

FIG - 7: Periderm tissue    

 

 

Figure -8: Liberiform fiberes and Parenchyma cells      

Fi: Fiberes; Pa: Parenchyma Cells; Pe: Periderm - fragment

 

Figure 9: Vessel elements

 

 

Figure 10: Liberiform fiberes

Fi: Fiberes; Pe: Perforation; Pi: Pits; VE: Vessel Elements

 

Figure 11 and 12: Vessel elements showing lateral

Pa: Rectangular Parenchyma cells; Pe: Perforation; Pi: Pits;

VE: Vessel Elements                                      

RESULT AND DISCUSSION:

Powder Microscopic Studies:

The plant parts - leaf, petiole, stem and root were made in to coarse powder. The powder was immersed in warm water to remove the air bubbles in the powder. The powder was the stained with dilute safranin, washed the excess of safranin and mounted in dilute glycerine for microscopic observation. Small fragments epidermal peelings were seen in the powder. The adaxial epidermal peeling appears in surface view (Fig: 1). The epidermal cells are polyhedral in outline with straight anticlinal walls. The walls are thick. In the cell lumen these are minute thin parallel cuticular striatious (Fig: 2).

 

The abaxial epidermis is also seen in surface view. The epidermis is densely stomatiferous. The stomata are narrowly elliptical. The stomata are cyclocytic type. The stomata are surrounded by four or five subsidiary cells (Fig: 3). Calcium oxalate crystals are seen scattered in the power (Fig: 4). The crystals are prismatic type. They are also dimses which are spherical bodies with spiny surface. Peltate Scales: (Fig: 5). Peltate scales are unique type of epidermal structures they are circular, thin plates comprising several-long pear - shaped scales other lateral to form a serrate circular plate (Fig: 6).

 

Small pieces of periderm:

Broken and isolateral pieces of periderm is often seen in the powder (Fig: 7). The periderm cells are narrowly rectangular and are arranged in compact parallel radial files. The cells are thin walled and straight the cells are phellem portion of the periderm. Fibres (Fig: 8). Liberiform fibres are abundant in the powder. They are long narrow, needle line cells with thick walls, reduced cell lumen and tapering ends. Lateral walls pits are not evident in the fibres. The fibres are 450 - 650 µm long and 20µm thick.

 

Vessel elements:

(Fig 9.10): Vessel elements common in the powder they are wide short cylinders (Fig: 11 and12). The vessel elements have prominent circular, much seriate alternate pits on the lateral walls (Fig: 12). Wide perforatous are seen on the end walls. The perofratious horizontal in orientation in short wide vessel elements (Fig: 7, 8). The peroforatious are narrow, elliptical and oblique in the case long narrow vessel elements range in length from 300 - 400µm they are 20 -70µm wide.

 

CONCLUSION:

The present investigation it can be concluded that the powder microscopically studies of Thespesia populnea yielded a qualitative and quantitative parameters or standards that can serve as an important possible sources of information to ascertain the identity and to determine the quality and purity of the plant material. These information will also be helpful to differentiate Thespesia populnea from the closely related other species and varieties of Thespesia.

 

ACKNOWLEDGEMENT:

I express my sincere gratitude to Prof. P. Jeyaraman, Director, Plant Anatomy Centre, West Tambaram, Chennai - 600 045 to carry out the anatomical work. Dr. M. Senthilkumar, Assistant Professor and Research Coordinator, PG Research Department of Botany, Vivekanandha College of Arts and Sciences for Women (Autonomous), Elayampalayam–637 205, Tiruchengode, Tamil Nadu, India for his critical analysis.

 

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2.        Sathyanarayana, T., Sarita, T., Balaji, M., Ramesh, A., Boini., M.K. Antihyperglycemic and hypoglycemic effect of Thespesia populnea fruits in normal and alloxan–induced diabetes in rabbits. Saudi Pharmaceutical Journal. 2004; 12: 107-111.

3.        Ilavarasan, R., Vasudevan, M., Anbazhagan S., Venkataraman S. Antioxidant activity of Thespesia populnea bark extracts against carbon tetrachloride induced liver injury in rats. J. Ethanopharmacology. 2003; 87: 2-3: 227-230.

4.        Ilavarasan, R., Vasudevan, M., Anbazhagan S., Venkataraman S., Sridher, S.K. Hepatoprotective activity of Thespesia populnea bark extracts against carbon tetrachloride-induced liver injury in rats. Natural Product Sciences. 2003; 9: 83-86.

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9.        Murthy, R.S.R., Basu, D.K., Murti, V.V.S. Antifertility activity of (+) gossypol. Current Sciences. 1981; 50: 64 - 66.

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13.      Sass, J.E. 1940. Elements of Botanical Microtechnique. McGraw Hill Book Co; New York. pp.222.

14.      Johansen, D.A. 1940. Plant Microtechnique. Mc Graw Hill Book Co; New York. Pp.523.

15.      O’Brien, T.P; Feder, N. and Mc Cull, M.E. 1964. Polychromatic Staining of Plant Cell walls by toluidine blue-O.protoplasma; 59:364 - 373.

16.      Easu, K. 1979. Anatomy of seed Plants. John Wiley and sons. New York. pp. 550.

 

 

 

 

Received on 03.07.2019         Modified on 31.07.2019

Accepted on 18.08.2019       ©A&V Publications All right reserved

Res.  J. Pharmacognosy and Phytochem. 2019; 11(3):189-192.

DOI: 10.5958/0975-4385.2019.00032.3