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Micromorphological and Phytochemical Studies on Cleome Rutidosperma Linn

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Micromorphological and Phytochemical Studies on Cleome Rutidosperma Linn Journal of Advances in Biology & Biotechnology 11(3): 1-8, 2017; Article no.JABB.31028 ISSN: 2394-1081 SCIENCEDOMAIN international www.sciencedomain.org Micromorphological and Phytochemical Studies on Cleome rutidosperma Linn. K. Okonwu 1* , C. Ekeke 1 and S. I. Mensah 1 1Department of Plant Science and Biotechnology, Faculty of Science, University of Port Harcourt, P.M.B. 5323, Choba, Rivers State, Nigeria. Authors’ contributions This work was carried out in collaboration between all authors. Authors KO and CE designed the study, performed the statistical analysis, wrote the protocol, managed the literature searches and wrote the first draft of the manuscript. All authors read and approved the final manuscript. Article Information DOI: 10.9734/JABB/2017/31028 Editor(s): (1) Mohammad Arif, Department of Plant Pathology, Kansas State University, Manhattan, Kansas, USA. Reviewers: (1) Abdullahi M. Nuhu, Kaduna Polytechnic, Kaduna, Nigeria. (2) Preeya P. Wangsomnuk, Khon Kaen University, Thailand. Complete Peer review History: http://www.sciencedomain.org/review-history/17534 Received 16 th December 2016 Accepted 12 th January 2017 Original Research Article th Published 16 January 2017 ABSTRACT The study was carried out to determine the micromorphological structure and quantify the phytochemical components of Cleome rutidosperma Linn. The study revealed the presence of five stomatal types (anomocytic, staurocytic, tetracytic, anisocytic and isotricytic) and polar contiguous stomata on both abaxial and adaxial surface of the leaf of C. rutidosperma . The shapes of the adaxial epidermal cells are relatively regular while the abaxial epidermal cells are irregular. There are fewer stomata on the adaxial epidermal surface than the abaxial surface. The range of the stomatal index (SI) on the adaxial and abaxial surfaces is between 3.85-20.0 and 63.64-84.21 respectively. Cleome rutidosperma has eglandular trichomes which occur on the leaf surfaces and stem of the plant; these trichomes are disrriate or trisariate with multicellular base. The petiole has five U-shaped free open collateral vascular bundles. The midrib is U-shaped adaxially and contains 5-free vascular bundles which formed a semicircle. The adaxial and abaxial epidermis have single layer of cell with 4-5 and 4-6 layers of parenchymatous cells, respectively. The upper parenchyma comprised 3-4 layers of cell while lower parenchyma has 4-7 layers of cells. The fruit stalk comprised 7-vascular bundles in a concentric ring. The cuticle is undulated with one-layer of epidermis and 3-4 layers of parenchymatous cells. The stem is pentagonal with 18-20 vascular _____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected]; Okonwu et al.; JABB, 11(3): 1-8, 2017; Article no.JABB.31028 bundles in a concentric ring. The epidermis has 1-layer of cell, collenchymatous cells 1-3 layers and parenchyma cells 1-4 layers but up to 6-10 layers the angles or protruded ends. The quantitative analysis of the phytochemicals showed that the C. rutidosperma contains 1.64% alkaloid, 2.84% flavonoid, 0.145% tannin, 3.25% saponin, 30 mg/kg cyanogenic glycoside, 48.8 mg/g oxalate while phytate was not detected. These micromorphological information recorded in this work can be used to distinguish C. rutidosperma from other members of the genus and are therefore of taxonomic value. Also, the presence of these phytochemicals could account for its use by the local communities in south-east Nigeria for the treatment of ear infection and other illnesses. Keywords: Anatomy; Cleome; phytochemical; stomatal index; trichome. 1. INTRODUCTION the extract from the leaf of C. rutidosperma calms irritation and itching in the ear, this Cleome rutidosperma Linn. belongs to the family experience has made the indigene of Ogoni to Cleomeceae. It is native to Nigeria and has been describe C. rutidosperma as "hospital too far" reported to be among the 50 species of Cleome because of the relieve it provides. A similar claim occurring in Africa [1]. A plant with spreading and was also reported in India that C. rutidosperma prostrate branches and leaves divided in 3 relieve ear pain and skin diseases [8]. leaflets. Flowers are small, violet-blue, 5 mm long, borne on long stalks from the leaf axil of the Preliminary phytochemical screening of the flowering stems. Fruit is cylindrical and curved, aqueous extract of C. rutidosperma showed the about 5 cm long, with numerous round, reddish- presence of tannins, saponins, flavonoids and brown to black seeds, 1.5 mm diameter with carbohydrates [10]. Similar phytochemical barred surfaces. C. rutidosperma is an annual screening carried out on the seed flours of species reproducing solely by seed. In Africa, C. rutidosperma revealed the presence of within its native distribution range, flowering and alkaloids, steroids, tannins, flavonoids, cardiac fruiting plants of C. rutidosperma can be found glycosides, pentose, and reducing sugars [11]. throughout the year, although most abundantly in This promising plant, C. rutidosperma, if well the rainy season [2]. It is a common weed with researched will clear the doubt of its inherent economic impacts in a wide range of crops, potential in curing most ailments. This study where its scrambling habit smothers and stunts focuses on the micromorphological and young crop plants. It is an environmental weed in quantification of some of the phytochemical disturbed ground, roadsides, gardens, and component of C. rutidosperma otherwise abandoned land as well in natural and semi- considered by many as weed. The result of the natural coastal forest where it has the potential to study will broaden the taxonomic knowledge outcompete native vegetation [3,4]. base and probably provide information for the C. rutidosperma spreads by seeds. Seed pharmaceutical industries to improve upon. dispersal is myrmecochorous, ants being attracted to the seeds by the fatty elaiosome[5]. 2. MATERIALS AND METHODS In Malaysia, C. rutidosperma is planted around field edges as part of insect control programs 2.1 Sources Plant Material [2]. Fresh plant materials of C. rutidosperma used for The young shoots and leaves of C. rutidosperma this study were collected from University of Port are eaten as a cooked vegetable or added to Harcourt Park, properly identified, authenticated soup in most part of Africa and Asia [6,7]. In and deposited in the University of Port Harcourt Ghana, Gabon and DR Congo, the leaf sap is Herbarium with reference number UPH/V/1254. applied to cure earache and deafness. In Ghana, the leaf extract of C. rutidosperma is used to 2.2 Anatomical Studies treat irritated skin and in Nigeria it is used to treat convulsions [1]. The plant is used as antimalarial The central portions of the matured leaf were by traditional healers in Cameroon. The plant is peeled, stained with 1% safranin O and in slides used in the treatment of paralysis, epilepsy, with glycerin. Specimens for anatomical analysis convulsion and spasm [8]. The plant is frequently were obtained fresh from matured plants and used in traditional medicine [7,9]. Ethno-botanical fixed in FAA for 12 hrs. They were transferred to survey conducted in Ogoni, Nigeria claimed that 50% and 70% ethanol and kept at room 2 Okonwu et al.; JABB, 11(3): 1-8, 2017 ; Article no.JABB .31028 temperature un til required. The petiole, midrib Where: and fruit stalk were hand sectione d using sharp razor blades [12, 13]. The sections were stained A = weight of filter and residue and in 1% Safranin red for two minutes, counter B = weight of empty filter paper stained Alcian blue and mounted on a slide. Thereafter, the slides viewed and phot ographed 2.5 Determination of Flavonoid by Bohm with LeitzDiaplan photomicroscope fitted with and Kocipal-Abyazan (1994) Leica WILD MPS 52 camera. The terminology used in respect to stomatal complex followed 10 g of the plant sample was extracted Malvey [14]. repeatedly with 100 mL of 80% aqueous methanol at room temperature. The whole 2.3 Determination of Tannin solution was filtered through Whatman filter paper No 42 (125 mm). The filtrate was later The percentage composition of tannin in the transferred into a crucible and evaporated into plants was determined using the methods of dryness over a water bath and weighed to a AOAC (1980) with some modifications. Folin - constant weight. Denis reagent and saturated sodium carbonate were prepared in accordance with the procedure to analyze the tannin content. Sta ndard solution of tannic acid was freshly prepared by dissolving 10 mg of tannic acid in 100 mL water. A series of Where; tannic acid standards were prepared in the range of 0-2.5 mL, aliquots in 25 mL volumetric flasks, A = weight of flask and sample residue and then added to 1.25 mL Folin-Denis reagent and B = weight of empty flask 2.5 mL sodium carbonate solution. The mixture was made up to the volume and the colour was 2.6 Determination of Phytate measured after 30 min at 760 nm using a spectrophotometer (Perkin Elmer). The samples 0.1 g of the plant was extracted with 100 m L of were prepared by boiling 1 g of their dried 0.5 M HCl for 2 hours and filtered. 15 m L of the powder in 80 mL of water for 30 min. The aliquot was neutralized with 0.5 M NaOH, made samples were cooled, transferred into a 100 mL slightly acidic with 0.17 M HCl and diluted to 50 volumetric flask and diluted to mark. The solution mL with distilled water. 4 mL of 0.25% w/v FeCl 3 was filtered to get a clear filtrate and analyzed as (0.25 g in 100 mL) added to 10 m L aliquot in in the standard. Tannin content was determined centrifuge tube, heated for 15 minutes at 100 °C, by a tannic acid standard curve and expressed cooled, centrifuged and supernatant discarded.
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