Morphological and Microscopical characters of Eleusine indica (L.) Gaertn.

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1 Universities Research Journal 2011, Vol. 4, No. 1 Morphological and Microscopical characters of Eleusine indica (L.) Gaertn. Khin Ohnmar Saw Abstract Eleusine indica (L.) Gaertn. locally known as sin-ngo-myet was collected from North Dagon Myothit Township, Yangon Division and the whole plant was studied in this research. According to the morphological characters present in the vegetative and reproductive parts of the plants, it has been identified by referring available literature. Eleusine indica (L.) Gaertn. belongs to the family Poaceae, naturally grown in fields and open grounds and found abundantly during the rainy season. The microscopical characters of fresh leaves, culms, roots and the dried powder of the whole plant were examined to asceratin its identification. The epidermal cells are arranged in parallel row. The bulliform or motor cells and two types of vascular bundles are observed in transverse section of lamina. The vascular bundles are scattered, except in the center of the culm in transverse section. Key words : sin-ngo-myet, culm, lamina, leaf-sheath Introduction One of the traditional medicinal plants namely Eleusine indica (L.) Gaertn. which belongs to the family Poaceae was selected in this study, The plant is a common herbage with long, narrow leaves and tubular culm, including cereals, bamboo, sugarcane, fodder grass, goose grass, wire grass etc. (Hundley, 1987). Family Poaceae is the largest of the world flora and contain a very wide range of chemical constituents. However, a large proportion of chemical work has been devoted, foodstuffs, starch, sugars and volatile oil (Trease and Evans, 2002). Eleusine indica (L.) Gaertn. is a worldwide weed of the tropics and one of the serious weedy grasses of the world. This plant is locally known as sin-ngo-myet. Kress et al., (2003) mentioned that this plant as myet-thakwa, se-gwa, sin-myet. Owing to the rapidity with which it occupies distributed ground especially useful though it is annual as soil binder activities (Bor, 1960). The plants are used for diaphoretic, febrifuge (antipyretic) and asthma (Nagathein, 1977). The whole plant, but more especially the root is considered diaphoretic, used for liver complaints and convulsion as antipyretic (Kirtikar and Basu, 1935; Assistant Lecturer, Department of Botany, Yangon University of Distance Education

2 226 Universities Research Journal 2011, Vol. 4, No. 1 Chopra, 1956). The juice of the leaves is given to a woman after child-birth to bring about the discharge of after birth (Burkill, 1935). Eleusine indica (L.) Gaertn. is an annual, erect, tufted, branched and glabrous grass. The culms are slender, terete and have nodes and internodes. The lamina arises from the nodes above leaf-sheath that overlaps and surrounds the culms. Spikes are terminal, the spikelets are numerous, two seriate. The florets are bisexual, the stamens 3, the ovaries ellipsoid, and the lodicules 2. In the present research, the morphological characters and microscopical characters of leaves, culms, roots and the dried powder of whole plant of Eleusine indica (L.) Gaertn. were studied to ascertain the identification of the specimen. Materials and Methods In this research, the specimens were collected from North Dagon Myothit Township during flowering and fruiting period. After the collection, the vegetative and floral parts of fresh specimens were used to identify the specimen. Taxonomic status of this plant was verified with the help of literatures (Hooker, 1894; Hitchock, 1950; Bor, 1960; Backer, 1963; Hafliger et al., 198l). All necessities were documented by photographs and drawings. Herbarium specimens were also prepared and kept in the herbarium, Department of Botany, Yangon University. The collected plants were washed, cut into small pieces and dried under shade, after which were powdered with grinding machine. The powder was kept in the airtight container until required for further experiments. For the microscopical studies, free hand sections of the fresh lamina, leaf-sheath, midrib, culm and root of Eleusine indica (L.) Gaertn. were prepared by using a razor blade. Microscopical characters were studied as determined by (Metcalfe and Chalk, 1950; Pandey, 1993). The stomatal index in leaves was determined by the method of (Wallis, 1955; Trease and Evans, 1978). The diagnostic characters of the powder were made by using the powder of the whole plant. The powder was cleared by chloral hydrate solution on a glass slide and observed under the microscope. N/50 iodine solution was used for the examination of starch. All lignified tissues were tested with 1% Phloroglucinol solution (B.P) followed by concentrated hydrochloric acid.

3 Universities Research Journal 2011, Vol. 4, No Results Morphological Characters of Eleusine indica (L.) Gaertn. Annual grasses, slender, erect, tufted. Culms slender, mm in length, mm in diameter, terete, branched, the nodes mm long, the internodes mm long. Leaf-sheaths mm long, mm wide, coriaceous on adaxial surface, glabrous on the abaxial surface, short-ciliate along the margins, ligules membranous acute, bearded at the mouth; lamina linear, mm long, mm wide, acuminate at the apex, entire at the margin, with long hairs on the adaxial surface. Inflorescences dichotomously forking adaxial panicles, composed of 2-7 digitately arranged spike-like racemes, all of them borne together, sometimes one of them attached at the lower node, mm long, mm wide, acuminate at the apex, primary peduncles straight, glabrous, the secondary peduncles continuous, slender. Spikelets lanceolate, mm long, mm wide, awnless, laterally compressed, pedicellate; pedicels mm long, scabrescent, the rachilla jointed above the glumes and between the florets, 2 to many flowered; glumes unequal, dioecious, the lower glumes lanceolate, mm long, 0.4-o.6 mm wide, awnless, membranous or coriaceous, acuminate at the apex, distinctly 1-nerved, scabrescent on the mid-nerve, the upper glumes elliptic, mm long, mm wide, awnless, coriaceous, acuminate at the apex, distinctly 4-nerved, scabrescent on the mid-nerve on the outer surface, glabrous on the inner surface, the lower florets the longest and the upper one progressively smaller. Florets lanceolate-oblong, mm long, mm wide, awnless, acute at the apex, sessile, bisexual, the lemma broadly ovate, mm long, mm wide, awnless, coriaceous, acute at the apex, distinctly 1-nerved, scabrescent on the midnerve, glabrous, the paleas elliptic, mm long, mm wide, awnless, acute at the apex, distinctly 2-nerved, lateral nerve not continuous to the apex, ciliate at the margins, glabrous, the lodicules 2, ob-conical, the stamens 3, filaments slender, short, mm long, anthers oblong, mm long, the ovaries ellipsoid, mm long, the styles 2, mm long, the stigmas 2, plumose, mm long, white. Habit, inflorescences and parts of spikelets are as shown in Fig. (1, 2, 3).

4 228 Universities Research Journal 2011, Vol. 4, No. 1 Microscopical characters of Eleusine indica (L.) Gaertn. Lamina In surface view, the epidermal cells of both surfaces with sinuous anticlinal walls are arranged in parallel rows. The cells are of two types; long and short cells. The short cells mostly solitary but occasionally in pairs, both over and between the veins, abundant, the intercostals short cells, filled with silica bodies. Long cells, between the veins with thin to moderately thick sinuous walls, vertically elongated. Stomata present on both surfaces. The stomata are graminaceous type. Stomata with markedly triangular subsidiary cells. In transverse section, the cuticle thin layered is 0.55 μ thick. The epidermis is one layered thick. The upper epidermal cells are rectangular in shape. The lower epidermal cells are similar to upper epidermal cells. One large vascular bundle alternate with three small vascular bundles. The ribs arched over vascular bundles. The layers of sclerenchymatous cells 2-3 cells thick, 8-19 cells wide over and under large vascular bundles and 2-3 cells thick, 7-9 cells wide over and under small vascular bundles. The vascular bundles are circular in outline structure. The bundle sheath of small bundle is single layered and parenchymatous but that of larger bundles are double layered with the outer parenchymatous and the inner sclerenchymatous layers. The bulliform cells are present at the furrows of the upper epidermis, oval or circular in shape and distinct. Xylem found towards upper surface and phloem towards lower surface. Xylem composed of vessel elements, fibers and xylem parenchyma. Phloem composed of sieve tube elements, companion cells and phloem parenchyma. The vessels present are 2-4 in each bundle while the phloem consists of cells. These characters are the same in all the apical, middle and basal regions but different in sizes. The microscopical characters of lamina are as shown in (Fig. 4-6). he measurement of cells in surface view and transverse section of lamina are given in Table (1-2).

5 Universities Research Journal 2011, Vol. 4, No Table 1. The measurement of cells in surface view of lamina Characters Length x width (μ) Upper epidermis (short cells) x Upper epidermis (long cells) x Lower epidermis (short cells) x Lower epidermis (long cells) x Stomata x Stomata Index Table 2. The measurement of cells in transverse section of lamina Characters Epidermis Vascular bundle Phloem Metaxylem Protoxylem Bundle sheath cell Basal region Middle region Apical region Length x width (μ) x x x x x x Bulliform cell x Length x width (μ) x x x x x x x Length x width (μ) x x x x x x x

6 230 Universities Research Journal 2011, Vol. 4, No. 1 Midrib In surface view, the epidermal cells are rectangular in shape, compact and parallel. The cells are two types; long and short. The anticlinal walls are sinuous. Graminaceous types of stomata are present. In transverse section, the cuticle is about 0.28 μ thick present on outer surface only. The epidermal cells are rectangular in shape, compactly arranged. The parenchymatous cells are hexagonal to polygonal in shape, about 6-8 layers at the basal region, 3-5 layers at the middle region and 2-3 layers at the apical region. The vascular bundles are closed collateral type, each circular, have outer and inner bundle sheaths. Outer sheath composed of parenchymatous cells, cells at the basal region, cells at the middle region and cells at the apical region, complete, inner sheath sclerenchymatous cells, cells at the basal region, cells at the middle region and cells at the apical region, complete. The bundle with layers of sclerenchyma, about 4-6 cells high and cells wide at the basal region, 3-5 cells high and cells wide at the middle region and 2-3 cells high and cells wide at apical region. Phloem lies towards outer surface. Number of vessels 2-5 in each bundle, xylem composed of vessel elements, fiber and xylem parenchyma. Number of phloem cells in each bundle, phloem composed of sieve tube elements, companion cells and phloem parenchyma. The microscopical characters of midribs are as shown in Fig. (7-12). The measurements of cells in transverse section of midrib are given in Table (3).

7 Universities Research Journal 2011, Vol. 4, No Table 3. The measurement of cells in transverse section of midrib Characters Epidermis Parenchyma Vascular bundle Phloem Metaxylem Protoxylem Basal region Middle region Apical region Length x width (μ) x x x x x x Length x width (μ) x x x x x x Length x width (μ) x x x x x x Leaf-sheath In surface view, the epidermal cells of both surfaces are dissimilar in shape and size. The cells are in parallel rows, rectangular and compactly arranged. Stomata are present on the outer surface and rare on the inner surface when present. The stomata are graminaceous type. In transverse section, the cuticle is present on both surfaces. Cuticle layer is thin on the outer surface about 0.28 μ thick and thicker on the inner surface about 1.1μ. Both inner and outer epidermal cells one layered, compact, anticlinal walls straight, periclinal walls convex but the shape of the inner epidermal cells are barrel-shaped and that of the outer epidermal cells are rectangular. The ground parenchymatous cells 4-12 layers, compact, thin walled, hexagonal in shape, variable in size. The vascular bundles are scattered, closed collateral type, oval to circular, bundle sheath complete, and parenchymatous cells cells. The number of vessels 4-7 in each bundle, xylem composed of vessel elements, fibers and xylem

8 232 Universities Research Journal 2011, Vol. 4, No. 1 parenchyma. The number of phloem cells in each bundle, phloem composed of sieve tube element, companion cells and phloem parenchyma. The microscopical characters of the leaf-sheath are shown in Fig. (13-15). The measurement of cells in transverse section of leaf-sheaths is given in Table (4). Table 4. The measurement of cells in transverse section of leaf-sheath Characters Length x width (μ) Epidermis (inner surface) x Epidermis (outer surface) x Parenchyma x Vascular bundle x Phloem x Metaxylem x Protoxylem x Culm In surface view, the cells are rectangular, compact, parallel, elongated along the length of the culm. The stomata rare, graminaceous when present. In transverse section, the culms are about 3-4 mm in diameter, oval in outline. The cuticle layer is thin, 0.44 μ thick. The epidermal cells are one layered, rectangular in shape and compactly arranged. The anticlinal walls straight, the periclinal walls convex. The ground tissue consists of two type of cells, sclerenchymatous and parenchymatous. The sclerenchymatous cells at the periphery, 4-8 layers, thin walled. The parenchymatous cells both outside and inside the sclerenchymatous layers, oval or rounded, cells compact, intercellular spaces present. The vascular bundles are scattered, devoid in the center of culm. Smaller bundles nearer the periphery. The bundle oval or rounded in outline, bundle sheath complete, sclerenchymatous, number of sclerenchymatous cells around each bundle. Xylem towards the centre, phloem towards the epidermis. Xylem composed of vessel elements, fiber and xylem parenchyma. The vessels present are 2-4 while the phloem consist of are

9 Universities Research Journal 2011, Vol. 4, No cells in each bundle. Phloem composed of sieve tube elements, companion cells and phloem parenchyma. The microscopical characters of clum are as shown in Fig. (16-18). The measurement of cells in the transverse section of culm is given in Table (5). Table 5. The measurement of cells in transverse section of culm Characters Length x width (μ) Epidermis x Parenchyma x Vascular bundle x Phloem x Metaxylem x Protoxylem x Root In surface view, the epiblema cells are rectangular in shape, regular or irregularly arranged, longitudinally elongated, compact, the anticlinal walls straight and smooth. In transverse section, the roots are about mm in diameter, circular in outline. The epiblema (piliferous layer) with numerous unicellular root hairs. The cells are regularly or irregularly arranged and rectangular to nearly polygonal in shape, the anticlinal walls straight, the periclinal walls convex. Hypodermis lies below the epiblema, one layered thick, parenchymatous and the cells are polygonal in shape and arranged irregularly. Ground tissue is thin walled, parenchymatous, consists of cortex, endodermis, pericycle, vascular bundle and pith. The cortex consists of 6-7 layered, polygonal parenchymatous cells, with intercellular spaces between them. Outer and inner ground tissues are separated by the endodermis and pericycle. The endodermis is the innermost layer of the cortex and forms a definite ring, one layered, cells parenchymatous. The cells are ellipsoidal to rectangular in shape, cell walls thin. The pericycle layer is lying internal to the endodermis, ring-like, one layered, cells parenchymatous, irregularly rectangular. The vascular bundles are radial type, polyarch, compactly arranged, xylem strands alternate with phloem

10 234 Universities Research Journal 2011, Vol. 4, No. 1 strands. Xylem and phloem are oval to round in outline, xylem cells are 3-7 in each strand, exarch, protoxylem occurring near the periphery and metaxylem inwards. Xylem is composed of vessels, tracheids, fibertracheids, fiber and xylem parenchyma. Phloem cells are in each strand; phloem is composed of sieve-tube elements, companion cells and phloem parenchyma. The mass of parenchymatous cells in the centre is the pith. The piths are made up of parenchymatous, cells, oval to rounded in shape and larger towards the centre. The microscopical characters of roots are as shown in (Fig ). The measurements of cells in transverse section of root are given in Table (6). Table 6. The measurement of cells in transverse section of root Characters Length x width (μ) Epiblema x Hypodermis x Cortex x Endodermis x Pericycle x Pith x Xylem x Phloem x Microscopical character of powdered the whole plant of Eleusine indica (L.) Gaertn. In this investigation, fragment of epidermal cell with stomata, fiber, tracheids, fiber-tracheids, spiral vessel and pitted vessel are observed. The sensory characters of the powder of Eleusine indica (L.) Gaertn. were given in Table (7) and their microscopical characters in Fig. (21).

11 Universities Research Journal 2011, Vol. 4, No Table 7. Sensory characters of powder of whole plant Sensory Characters Colour Odour Taste Texture Whole Plants Greenish Slightly aromatic Bitter Granular Fig. 1 Habit Fig. 2 Inflorescence

12 236 Universities Research Journal 2011, Vol. 4, No. 1 Fig. 3 Parts of Spikelets

13 Universities Research Journal 2011, Vol. 4, No Fig. 4 Surface view of upper epidermal cells showing Graminaceous type of stomata (X100) Fig. 5 Surface view of lower epidermal cells showing abundant stomata (X100) Fig. 6 Transverse Section of lamina

14 238 Universities Research Journal 2011, Vol. 4, No. 1 Fig. 7 Upper surface view of midrib (X100) Fig. 8 Lower surface view of Midrib (X400) Fig. 9 Transverse section of midrib (Apical region) Fig. 10 Transverse section of midrib (Middle region) Fig. 11 Transverse section of midrib (Basal region) Fig. 12 Transverse section of midrib showing vascular bundle

15 Universities Research Journal 2011, Vol. 4, No Fig. 13 Inner surface view of leaf-sheath (X100) Fig. 14 Outer surface view of leaf-sheath (X40) Fig. 15 Transverse section of leaf-sheath

16 240 Universities Research Journal 2011, Vol. 4, No. 1 Fig. 16 Surface view of culm (X100) Fig. 17 Transverse section of culm Fig. 18 Transverse section of culm showing vascular bundle Fig. 19 Surface view of root showing rectangular cells (X400)

17 Universities Research Journal 2011, Vol. 4, No Fig. 20 Transverse section of root Fig. 21 Microscopical character of powdered herbs (X400)

18 242 Universities Research Journal 2011, Vol. 4, No. 1 Discussion and Conclusion The plant Eleusine indica (L.) Gaertn. is usually found growing wild throughout the country during the rainy season. The morphological studies on both vegetative and reproductive parts as well as the microscopical examination of leaves, culms, and roots verified the plant to be Eleusine indica (L.) Gaertn. It is annual herb, erect, slender and tufted. The leaves are linear, parallel venation, distinguished into leaf-blades and leaf-sheaths. Ligules are present. The inflorescences are terminal spike. The spikelets are two to many flowered, awnless, and laterally compressed. The florets are awnless, bisexual, the lowest florets longest and upper one progressively smaller. The lodicules 2, ob-conical, the stamens 3, anther oblong, filament short, the ovary ellipsoid, styles 2, stigmas 2, plumose. All morphological characters of this plant are in agreement with those stated by Hooker (1894); Hitchock (1950); Bor (1960); Backer (1963); Hafliger et al., (1981). In microscopical studies, the epidermal cells of the surface view of lamina, midrib and leaf-sheaths are parallel rowed. The cells are of two types, long and short, walls sinuous. Bulliform cells are found in furrows of upper epidermis of lamina in transverse section. Epidermis is one layered thick. The cuticle is present on transverse section of midrib and leafshealth. The epidermal cells are one layered, compact, rectangular and that of inner surface of leaf-shealth barrel-shaped. Close collateral type of vascular bundles are found in all parts of leaf. In surface view of roots, the epiblema cells are rectangular. In transverse section of roots, the epiblema with numerous unicellular root hairs, cells are rectangular to nearly polygonal in shape. The ground tissue is thin-walled, parenchymatous, composed of cortex, endodermis, pericycle and pith. Vascular bundles are polyarch, radial type. In surface view of culm, the epidermal cells are elongated along the length of culm. In transverse section, the cuticle thin, epidermal cells are compact and rectangular in shape. Vascular bundles are scattered, but devoid in the center of culm. Fragment of epidermal cells with stomata, fibers, tracheids, fiber-tracheids, spiral vessels and pitted vessels are observed in the microscopical characters of powdered samples. These characters are agreement with Metcalfe & Chalk, (1950); Wallis, (1955); Esau, (1965); Pandey, (1993) and Sundara rajan, (2000). The sensory characters were found to be greenish, slightly aromatic, bitter, granular and these characters can be used for the identification and standardization of drug.

19 Universities Research Journal 2011, Vol. 4, No Acknowledgements I am greatly indebted to Dr. Yee Yee Wynn, Professor and Head, Department of Botany, Yangon University of Distance Education for allowing me to undertake this research. I am also greatly indebted to Dr. Thet Thet May, Professor and Head, Dr. Aye Pe, Professor Department of Botany, Yangon University for their kind help, invaluable advice for this research. I am also greatly indebted to Professor Naw Wah Wah Paw, Retired, Professor and Head, Department of Botany, Dagon University, Dr. Moe Moe Lwin, Lecturer, Department of Botany, Pathein University close guidance and numerous suggestions and comment offered during the research and Dr. Khin Myo Thwet, Lecturer, Department of Botany, Yangon University of Distance Education, for her kind help. References Backer, C.A., (1963). Flora of Java. Vol. 3. Noordhoof Griningen Company, The Netherland Bor, N. L., (1960). The Grasses of Burma, Ceylon, India and Pakistan. Pergamon Press, Oxford, London, New York and Paris Burkill, M. A., (1935). A Dictionary of the economic products of Malay Penisula. Vol. 1. London Chopra, R. N., (1956). Glossary of Indian medicinal Plants. Council of Scientific and Industrial Research, New Delhi Esau, K., (1965). Plant Anatomy. Second Edition, Printed in Japan, By Toppan Printing Company, Ltd Hafliger, E. and H. Schlog., (1981). Monocot Weeds 3 and Grass Weed 1, 2. CIBA. Geigy Ltd. Basl, Switzerland Hitchock, A. S., Mannual of The Grasses of The United States. Second Edition. Government Printing Office. United States Hooker, J. D., (1894). Flora of British India Vol.7. Reeve and Co., Ltd. London Hundley, H. G and Chit Ko Ko, (1961). List of Trees, Shrubs, Herbs and Principal Climbers etc. recorded from Burma. P Supdt. Govt. Printing and Stationary, Burma Kirtikar, K. R. and B. D. Basu, (1933). Indian Medicinal Plants. 2 nd ed., 49, Leader Road, Allalhabad, India Kress, J. and et al., (2003). A Checklist of Trees, Shrubs, Herbs and Climbers of Myanmar. Washington D.C Metcalfe, C. R. and L. Chalk, (1950). Anatomy of Monocotyledon Vol. 1. The Clarendon Press, Oxford

20 244 Universities Research Journal 2011, Vol. 4, No. 1 Nagathein, Ashin, (1977). Pon-pya-say-A-bidon. Vol. 1. Mingala Press. Yangon Pandey, B. P., (1993). Plant Anatomy. S. Chand & Company Ltd. Pamlvagar, New Delhi Sundara rajan, S. (2000). Plant Anatomy and Embryology. Anmol Publications. Ltd., Newdelhi Trease, G. E., and W. C. Evans, (1978). A Text Book of Pharmacognosy. 11 th Edition. Bailliere Tindall, London Trease G. E. and W. C. Evans (2002). Pharmacognosy. 15 th ed., Baillere Tindoll, London Wallis (1955). Text Book of Pharmacognosy. 3 th Edition., J & A Churchill Ltd., London