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Antifungal Properties and Phytochemical Screening of Extracts of African Basil (Ocimum Gratissimum

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Antifungal Properties and Phytochemical Screening of Extracts of African Basil (Ocimum Gratissimum AGRICULTURE AND BIOLOGY JOURNAL OF NORTH AMERICA ISSN Print: 2151-7517, ISSN Online: 2151-7525 © 2010, ScienceHuβ, http://www.scihub.org/ABJNA Antifungal properties and phytochemical screening of extracts of African Basil (Ocimum gratissimum L.) 1*Amadi, J.E, 1Salami, S.O and 2Eze, C.S 1Department of Plant Biology University of Ilorin, PMB 1515, Nigeria 2Department of Applied Biology and Biotech, Enugu State University, Enugu State [email protected] ABSTRACT A study was carried out to evaluate the antifungal properties of African Basil (Ocimum gratissimum L.) extracts on Aspergillus repens, Curvularia lunata and Fusarium moniliforme using the pour plate method. A phytochemical screening of the extracts was also carried out to determine the constituents of O. gratissimum. Water, ethanol and acetone were the extractants used. Results showed that radial growth in all the three test organisms was impaired by the addition of the extracts in the culture medium used. The test organisms differed in their reaction to the different extracts but on the whole, growth inhibition increased with the concentration of each extract. Phytochemical screening of the different extracts showed that O. gratissimum contains important compounds such as carbohydrates, reducing sugars, lipids, flavonoids, alkaloids, steroids, tannins and saponins. The significance of these results is discussed. Keywords: Ocimum gratissimum, antifungal, extracts, phytochemical, inhibition. INTRODUCTION compounds on the germination and growth of many Ocimum belongs to the family Lamiaceae, sub-family fungal pathogens have been reported. Ramos et al. Napetoideae of the tribe Ocimeae. Known species (2007) have reported components of the neem tree are O. gratissimum (African basil, tree basil), O. (Azadirachta indica) as promising candidates for the americanum (Hoary basil), O. basilicum (Sweet basil) control of Crinipellis perniciosa and Phytophthora O. campechiamum (Peruvian basil), O. minimum species. Ocimum extracts are used in traditional (Greek basil), O. kilimandscharicum (Camphor basil), medicine and have been shown to contain O.Citriodorum (Lemon Basil), O. tenuiflorum (Holy biologically active constituents that are insecticidal, Basil) and O. viride. O gratissimum is the most nematicidal, fungicidal and generally antimicrobial popular species in the genus. The genus Ocimum or (Morales and Simon, 1996). The essential oil, leaves, Mint, collectively called Basil, has long been seeds, flowers and roots of basils are used as acclaimed for its diversity. Ocimum constitutes medicine. between 50-150 species of aromatic annual and The objective of this study is to investigate the effect perennial herbs and shrubs from the tropical region of of O. gratissimum extracts on the growth of three Asia, Africa, Central and South America (Darrah, fungi namely Aspergillus repens, Curvularia lunata 1980). Basil originated from Iran, India and other and Fusarium moniliforme. Phytochemical screening tropical regions of Asia, having been cultivated there of the different extracts will also be carried out to for more than 5,000 years. Basil folklore is as determine their constituents. complex as its flavour and aroma. They are heat and drought tolerant (i.e. they do not require high MATERIALS AND METHODS humidity). Basil is a source of essential oils and Sample Collection and Extraction Procedures: aromatic compounds, culinary herb, and an attractive Fresh and matured leaves of O. gratissimum were fragrant ornamental. The seeds also contain edible collected from the Biological Garden of the University oils (Morales and Simon, 1996). of Ilorin. The leaf samples were collected in a sterile The presence of antifungal compounds, in higher polyethylene bag and taken to the herbarium of the plants, has long been recognised as an important Department of Plant Biology of the University for factor in disease resistance. Such compounds, being proper identification. The leaves were surface- biodegradable and selective in their toxicity, are sterilized using sodium hypochlorite and rinsed twice considered valuable for controlling some plant with sterile distilled water. Thirty (30g) grams of O. diseases (Siva et al., 2008). In addition, plant extracts gratissimum leaves were mashed in a sterile mortar might have inhibitors to enzymes from the invading using a sterile pestle. The resultant paste was pathogens, and the effects of different phenolic soaked for 24hr in 100ml of any of the three Agric. Biol. J. N. Am., 2010, 1(2): 163-166 extractants (water, acetone and ethanol) used. The Ocimum extracts to inhibit growth of organisms extract was filtered through sterile Whatman No. 1 differed with the extractant used, the organism itself filter paper and stored at 4oC for subsequent use. and the concentration of the extract used. On the Three different extracts were thus obtained in water, whole, A. repens was more sensitive to Ocimum acetone and ethanol. extracts than C. lunata and F. moniliforme. At the Pure cultures of the test organisms used (Aspergillus highest concentration tested in this study (40ml repens, Curvularia lunata and Fusarium moniliforme) extract: 120ml molten agar), all the Ocimum extracts were obtained from the Plant Pathology laboratory in completely inhibited growth in all the three organisms the Department of Plant Biology of the University of tested. At low concentrations, differences began to Ilorin. The Organisms were sub-cultured on potato show among the different extracts with reference to dextrose agar (PDA) plates and bottle slants and growth inhibition capabilities. At 20ml extract: 120ml stored in an incubator at 40C. molten agar concentration, the acetone extract was most effective against, A. repens in which complete Effect of Different Concentrations of Extracts on inhibition of growth was observed whereas a little Radial Growth of test Organisms: Different growth occurred in both C. lunata and F. moniliforme. concentrations of the extracts were obtained by At the same 20ml extract: 120ml molten agar amending 100ml of molten PDA with 5, 10, 20 and concentrations of the other extracts, A. repens was 40ml of the different extracts. The amended medium also completely inhibited while marginal growth was dispensed into sterile Petri plates and allowed to occurred in the other two test fungi. The aqueous solidify. Two perpendicular lines were drawn at the extract was the least effective in inhibiting growth of bottom of each plate to cross each other at the centre any the test fungi at all the concentrations tested in of the plate. Each plate was inoculated with any of this study. There was more growth in the control the test organisms. A 4-mm diameter mycelial disc of plates than in any of the test plates. C. lunata was the each of the test organisms was inoculated on each fastest growing organism among the three fungi used amended agar plate. Inoculated plates were for this study. incubated at 25±2oC and growth measured along the Phytochemical screening of the different Ocimum perpendicular lines. Daily radial growth of each test extracts revealed the presence of carbohydrates, organism in any of the test extracts was recorded for reducing sugars, lipids, flavonoids, alkaloids, 7 days. Each organism and extract was replicated steroids, tannins and saponins (Table 1). It was three times and the mean radial growth determined observed that the extractant used has effect on the for each test organism per test extract. final constituents of any plant extract because some Phytochemical Screening: Screening was carried constituents appeared only in one or two of the three out on all the Ocimum extracts to determine the extracts tested. active principles in them following the procedures of Table 1: Phytochemical Screening of Acetone Extract Sofowora (1993). Two (2) milliliters of each extract of Ocimum gratissimum was measured into a test tube for each of the tests and concentrated by evaporating the extractant in a Phytochemicals Acetone Extract trough. Tests were carried out for carbohydrates, Carbohydrate + reducing sugars, tannins, polyphenols, lipids, Reducing sugar + flavonoids, ketones, alkaloids, steroids and triterpenes. The screening was carried out in the Lipids + Department of Chemistry of the University of Ilorin, Alkaloids + Nigeria. Flavonoids + Steroids + RESULT Tanins + Effects of Extracts on Growth of Test Organisms: Saponins + Results of this study showed that the different extracts of O. gratissimum have inhibitory effects on Triterpenes - the growth of the test organisms. The potential of 164 Agric. Biol. J. N. Am., 2010, 1(2): 163-166 Plate 1: Mycelia and conidia of Curvularia lunata DISCUSSION plant extracts has been shown to be affected by the extractant, the extraction procedure and Results of this study show that all the three extracts concentration. Chloroform extracts of Passiflora of O. gratissimum exhibited varying degrees of edulis was reported (Johnson et al., 2008) to have antimicrobial activities against C. lunata, A. repens shown maximum efficacy against the tested bacteria. and F. moniliforme. All of these organisms are known Both benzene and petroleum ether extracts were to cause different diseases in plants, especially crop found to be ineffective in bacterial inhibition, due to plants. Thus, effective antimicrobial activities against the presence of less number of active compounds these organisms will be handy towards controlling the like saponins, steroids and alkaloids. Reid et al. diseases they cause in plants. Growth assessment
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