Effects of Hot and Cold Leaves Extracts of Azadirachta Indica on Xanthomonas Campestris
Keywords:
Antibacterial, Azadirachta indica, Xanthomonas campestris, Phytopathogenicity, extracts
Abstract
There is a worldwide interest in searching for safe and effective novel antibacterial compounds of plant origin for the control of plant pathogenic bacteria which is responsible for the great impact on the growth and productivity of agricultural crops. In this study, the effects of hot and cold leaf extracts of Azadirachta indica on Xanthomonas campestris was carried out. The organism was isolated from symptomatic pepper fruits and leaves using Yeast Dextrose Chalk Agar (YDCA). The isolate was characterized and identified using its colonial description, Gram staining and biochemical characteristics. The phytochemical constituents of the leaf extract of Azadirachta indica were determined quantitatively using spectrophotometeric method. The antibacterial activity of the leaf extracts was carried out using agar-well diffusion method. Tube dilution method was used to determine the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) using double-fold serial dilution at concentrations 31.25mg/ml to 500mg/ml. The phytochemical analysis of Azadirachta indica leaf extracts revealed the presence of tannins, flavonoid, alkaloids, steroids, cardiac glycosides, phenolics and saponins. The results of the present study revealed the pronounced activities of the leaf extracts against the tested organism, which differed significantly (p < 0.05) from ciprofloxacin. The activities of hot n-hexane (8.33mm), ethanolic (7.00mm), petroleum-ether (6.33mm) and aqueous (6.00mm) leaf extracts differed significantly (p < 0.05) from activities of cold ethanolic (22.67mm), n-hexane (20.67mm), petroleum-ether (19.33mm) and aqueous (8.33mm) leaf extracts. The results of the MICs and MBCs of the leaf extracts of Azadirachta indica showed that the cold extracts exhibited inhibitory and cidal activities against the organisms. This study recommends that cold extraction using organic solvents is suitable for studying the antibacterial activity of Azadirachta indica and Azadirachta indica leaf extracts could serve as alternative pesticide to plant diseases caused by Xanthomonas campestris.
References
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Atata, R., Sani A. and Ajewole, S.M. (2003). Effect of stem back extracts of Enantia Chloranta on some clinical isolates. Biokemistri 15: 84–92.
Baez, D.H., De los Ryos, C., Crescente, O. and Caserta, A. (1998). Antibacterial and chemical evaluation of Chromolaena moritziana. Journal of Ethnopharmacy 59: 203-206.
Bergsma-Vlami, M., Martin, W., Koenraadt, H., Teunissen, H., Pothier, J.F., Duffy, B. and Vandoorn, J. (2012). Molecular typing of Dutch isolates of Xanthomonas arboricola pv. pruni isolated from ornamental cherry laurel. Journal of Plant Pathology 94: 29–36.
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Hanife, O. and Ahmet, A. (2009). Antimicrobial activity of some water plants from the Northeastern Anatolian region of Turkey. Molecules 14:321–328.
Hufford, C.D., Tia, Y., Croom, E.M.J., Mohammed, I., Okunade, A.L., Clark, A.M. and Rogers, R.D. (1993). Antimicrobial compounds from Petalostemum purpureum. Journal of Natural Products 56: 1878–1889.
Iheukwumere, I., Uba, B.O. and Ubajekwu, C.C. (2012). Antibacterial activity of Annona muricate and Persea americana leaves extracts against ampicillin-resistant Staphylococcus aureus. Journal of Science Engineering and Technology 19: 10786–10798.
Iheukwumere, I.H and Umedum, C.U. (2013). Effects of Gossypium hirsutum leaf extracts on Gram negative bacteria isolated from the cervix of females with unexplained infertility. African Journal of Science 14 (1): 3261–3270.
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Jeyaseelan, E.C., Pathmanathan, M.K. and Jeyaderan, J.P. (2011). Inhibitory effect of different solvent extracts of Vitex negundo and Allium sativum on phytopathogenic bacteria. Archives of Applied Science Research 3: 1–8.
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Kotzekidou, P., Giannakidis, P. and Boulamatsis A. (2008). Antimicrobial activity of some plant extracts and essential oils against foodborne pathogens in vitro and on the face of inoculated pathogens in chocolate. Food Science and Technology 41:119–127.
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Ali, A., Haider, S.M., Hanif, S. and Akhatar, N. (2014). Assessment of the antibacterial activity of Cuscuta pedicellata. Academic Journal, 13: 430–433.
Almeida, N.F., Yan, S., Cai, R., Clarke, C.R., Moris, C.E., Schaad, N.W and Schuenzel, E.L. (2010). PAMDB, A multilocus sequence typing and analysis database and website for plant associated microbes. Phytopathology 100: 208–215.
Amtul, J.S. and Shakoori, A.R. (2014). The potential of (Azadirachta indica) based saponin as biopesticides for invitro insect pests cellulase (Beta-1,4-endoglucanase) enzyme inhibition and in vivo repellency on Tribolium casataneum. British Biotechnology Journal 4(8): 904–917.
Arunkumar, S. and Muthuselvan, M. (2009). Analysis of phytochemical constituents and antimicrobial activities of Aloe vera L. against clinical pathogens. World Journal of Agricultural Sciences 5: 572–576.
Atata, R., Sani A. and Ajewole, S.M. (2003). Effect of stem back extracts of Enantia Chloranta on some clinical isolates. Biokemistri 15: 84–92.
Baez, D.H., De los Ryos, C., Crescente, O. and Caserta, A. (1998). Antibacterial and chemical evaluation of Chromolaena moritziana. Journal of Ethnopharmacy 59: 203-206.
Bergsma-Vlami, M., Martin, W., Koenraadt, H., Teunissen, H., Pothier, J.F., Duffy, B. and Vandoorn, J. (2012). Molecular typing of Dutch isolates of Xanthomonas arboricola pv. pruni isolated from ornamental cherry laurel. Journal of Plant Pathology 94: 29–36.
Bishnu, J., Govind, P.S., Buddha, B.B., Megh, R.B., Dinita, S., Krishna, S., Janardhan, P. and Rajani, M. (2011). Phytochemical extraction and antimicrobial properties of different medicinal plants: Ocimum sanctum (Tulsi), Eugenia caryophyllata (Clove), Achyranthes bidentata (Datiwan) and Azadirachta indica (Neem). Journal of Microbiology and Antimicrobials 3: 1–7.
Biwas, Kausik, Ishita C., Ranajit, K.B. and Uday, B.P. (2002). Biological activities and medicinal properties of neem (Azadirachta indica). Current Science 82(11): 1336–1345.
Bottger, E.C. (1996). Approaches for identification of microorganisms. A.S.M News 62: 247–250.
Bull, C.T., De Boer, S.H., Denny, T.P., Firrao, G. and Takikawa, Y. (2008). Demystifying the nomenclature of bacterial plant pathogens. Journal of Plant Pathology 90 (3):403–417.
Cheesbrough, M. (2000). Medical Laboratory Manual for Tropical Countries Volume 2, 2nd edition. Cambridge University Press, Cambridge, Pp. 377.
Chinnappan, A. (2011). Antimicrobial potential and phytochemical screening of Andrographis affinis Nees- An endemic medicinal plant from India. International Journal of Pharmacy and Pharmaceutical Sciences 3: 157–159.
Corato, U., Maccioni, O., Trupo M. and Di Sanzo, G. (2010). Use of essential oil of Laurus nobilis obtained by means of a super critical carbondioxide technique against post harvest spoilage fungi. Contributions in Microbiology 29: 142–147.
Das, A.K. (2003). Bacterial plant pathology. Journal of Applied Horticulture 3: 52–60.
Davis, A.P., Rafael, G., Diane, M.B., Marku, S.R., Justin, M. and Neil, A.B. (2009). A global assessment of distribution, diversity, endemism and taxonomic effort in the Rubiaceae. Annals of the Missouri Botanical Garden 96: 68–78.
De, N. and Ifeoma, E. (2002). Antimicrobial effects of components of the bark extracts of neem (Azadirachta indica A. Juss). Journal of Technology Development 8: 2328.
Dixon, R.A. (2011). Plant pathology. Nature 411: 843–847.
Dwivedi, S. D., Sheeraz, A. and Wagay (2014). Antimicrobial activity of leaf extracts of Jurlnea dolomiaea plant against clinical and phytopathogenic bacteria. Chemical and Process Engineering Research 24: 2224–7467.
Dye, D.W., Bradbury, J.F., Goto, M., Hayward, A.C., Lelliott, R.A., Schrotch, M.N. (1980). International standards for naming pathovars of phytopothogenic bacteria and a list of pathovars names and pathogens. Review of Plant Pathology 59: 153–168.
Ebong, P.E., Atangwo, I.J., Eyong E.U. and Egbung, G.E. (2008). The antidiabetic efficacy of combined extracts from two continental plants: Azadirachta indica (A. Juss) (Neem) and Vernonia amygdalina (Del) (African bitter leaf). American Journal of Biochemistry and Biotechnology 4 (3): 239–244.
El. Mahmood, A.M. Ogbonna, O.B. and Raji, M. (2010). The antibacterial activity of Azadirachta indica (neem) with eye and ear infections. Journal of Medicinal Plants Research 4(14): 1414–1421.
Elphinstone, J.G. (2005). The current bacterial wilt situation: a global overview. In: Bacterial wilt disease and the Ralstonia solanacearum species complex. APS Press, St. Paul, Minnesota Pp. 9–28.
Favero, J.P., Courbeau, M., Nicolas, M., Benkirane, G., Trave, J.F.P., Dixon, P., Aucouterier, S., Rasheed, P. and Parker, J.W. (1993). Inhibition of human immunodeficiency virus infection by the lectin jacalin and by a derived peptide showing a sequence similar with Gp 120. Environmental Journal of Immunology 23: 179–185.
Ghoshal, S., Krishna, B.N. and Lakshmi, V. (1996). Anti-amoebic activity of Perperlongum fruits against Entamoeba histolytica. Journal of Ethno pharmacology 50: 167–170.
Gislene, G.E.N., Lacatelli, J., Paulo, C.F. and Giuliana, L.S. (2000). Antibacterial activity of plant extracts and phytochemicals on antibiotic resistant bacteria. Brazilian Journal of Microbiology 31: 247–256.
Green, S., Studholme, D.J., Laue, B.J., Dorati, F., Lovell, H., Arnold, D., Cottrell, J.E, Bridgett, S., Blaxter, M., Huitemast, Thwaites, R., Sharp, P.M., Jackson, R.W. and Kamoun, S. (2010). Comparative genome analysis provides insight into the evolution and adaptation of Pseudomonas syringae pathovar. aesuli on Aesulus hippocastanum. PLoS ONE, 5 (4): 1371.
Hammer, K.A., Carson, C.F. and Riley, T.V. (1999). Antimicrobial activity of essential oils and other plant extracts. Journals in Applied Microbiology 86(6): 985.
Hanife, O. and Ahmet, A. (2009). Antimicrobial activity of some water plants from the Northeastern Anatolian region of Turkey. Molecules 14:321–328.
Hufford, C.D., Tia, Y., Croom, E.M.J., Mohammed, I., Okunade, A.L., Clark, A.M. and Rogers, R.D. (1993). Antimicrobial compounds from Petalostemum purpureum. Journal of Natural Products 56: 1878–1889.
Iheukwumere, I., Uba, B.O. and Ubajekwu, C.C. (2012). Antibacterial activity of Annona muricate and Persea americana leaves extracts against ampicillin-resistant Staphylococcus aureus. Journal of Science Engineering and Technology 19: 10786–10798.
Iheukwumere, I.H and Umedum, C.U. (2013). Effects of Gossypium hirsutum leaf extracts on Gram negative bacteria isolated from the cervix of females with unexplained infertility. African Journal of Science 14 (1): 3261–3270.
Jackson, R.W. (2009). Plant pathogenic bacteria. Genomics and Molecular Biology 128: 521.
Jeyaseelan, E.C., Pathmanathan, M.K. and Jeyaderan, J.P. (2011). Inhibitory effect of different solvent extracts of Vitex negundo and Allium sativum on phytopathogenic bacteria. Archives of Applied Science Research 3: 1–8.
Kagbo, H.D. and Ejebe, D.E. (2010). Phytochemistry and preliminary toxicity studies of the methanol extract of the stem bark of Garcinia kola. International Journal of Toxicology 7 (2):1–16.
Kotzekidou, P., Giannakidis, P. and Boulamatsis A. (2008). Antimicrobial activity of some plant extracts and essential oils against foodborne pathogens in vitro and on the face of inoculated pathogens in chocolate. Food Science and Technology 41:119–127.
Kumar, V.S. and Navaratham, V. (2013). Neem (Azadirachta indica): prehistory to contemporary medicinal uses to humankind. Asian Pacific Journal of Tropical Biomedecine 3 (7): 505–514.
Lalitha, V., Raveesha, K.A. and Kiran, B. (2010). Antimicrobial activity of Solanum torvum swart against important seed borne pathogens of paddy Iranica Journal of Energy and Environment 1(2): 160–164.
Leben, C. and Whitmoyer, R.E. (1979). Adherence of bacteria to leaves. Canadian Journal of Microbiology 8: 896–901.
Letidal, E.T.M., El bushia, S.N. and Mahasin, A.N. (2010). The antibacterial, antiviral activities and phytochemical screening of some Sudaneses medicinal plant. European Asian Journal of Biological Science 4: 8–16.
Millikan, T.C., Suvimol, S., Veena, S.N. and Wandee, G. (2005). Antimicrobial effects of Thai medicinal plants against acne-producing bacteria. Journal of Ethnopharmacology 5:1–4.
Mole, B.M, Baltrus, D.A., Dangl, J.L. and Grant, S.R. (2007). Global virulence regulation networks in phytopathogenic bacteria. Trends in Microbiology 15: 363–371.
Muhldorfer, I. and Schafer, K.P. (2001). Emerging Bacterial Pathogens. Contributions in Microbiology 8: 186–196.
Natarajan, V., Veugopal, P.V. and Menon, T. (2003). Effect of Azadirachta indica (Neem) on the growth pattern of dermatophytes. Indian Journal of Medical Microbiology 21: 98–101.
Nwokeke, C. (2008). The efficacy of bitter kola as a potent antibiotic ion treating opportunistic infections associated with HIV. Nigerian Natural Medicine Development Agency (NNMDA). An interview by News Agency of Nigeria (NAN) P. 5.
Onocha, P.A. and Otunla, O.E. (2010). Biological activities of extracts of Pyenanthus angolensis (Welw) warb. Archives of Applied Science Research 2: 186–190.
Opara, E.U. and Wokocha, R.C. (2008). Biological control of bacteria plant pathogens. Agricultural Journal 3: 163–170.
Parekh, J., Nair, R. and Chanda, S. (2005). Preliminary screening of some folklore medicinal plants from Western India for potential antimicrobial activity. Indian Journal in Pharmacology 37: 408–409.
Perumal Samy, R. and Ignacimuthu, S. (2000). Antibacterial activity of some medicinal plants from Eastern Ghats, South India. Solai Bulletin Ethnopharmacology 72: 39–41.
Roberto, B. (2008). Plant- microbe interaction; infection and plant defense responses during plant-bacterial interaction. Research Project 8: 169–197.
Roberto, B. (2008). Plant-microbe interaction; infection and plant defense responses during plant-bacterial interaction. Research Project 8:169–197.
Santhosh, M.S., Hemshekhar, M., Sunitha, K., Devaraja, S., Kemparaju, K. and Vishwanath, B.S. (2011). An overview on genus Garcinia: phytochemical and therapeutical aspects. Phytochemistry Reviews 10(3): 325–351.
Satish, S., Raveesha, K.A. and Janardhana, G.R. (1999). Antibacterial activity of plant extracts of phytopathogenic Xanthomonas campestris pathovars. Letters of Applied Microbiology 28: 145–147.
Satish, S., Raveesha, K.A. and Janardhana, G.R. (1999). Antibacterial activity of plant extracts on phytopathogenic Xanthomonas campestris pathovars. Letters in Applied Microbiology 8: 145–147.
Scarbert, A. (1991). Antimicrobial properties of tannin phytochemistry. African Journal of Biotechnology 30: 3875–3883.
Soine, T.O (1994). Naturally occurring coumarins and related physiological activities. Journal of Ethnopharmacology 45:43–52.
Stackebrandt, E., Frederiksen, W., Garrity, G.M., Grimont, P.A.D., Kampfer P., Maiden, M.C.J., Nesme, X., Rossello-Mova, R. and Swings, J. (2002). Report of the ad hoc committee for the re-evaluation of the species definition in bacteriology. International Journal of Systematic Evolution in Microbiology 52: 1043–1047.
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Published
2018-04-04
How to Cite
E., E., & H., I. (2018, April 4). Effects of Hot and Cold Leaves Extracts of Azadirachta Indica on Xanthomonas Campestris. African Journal of Education,Science and Technology, 3(3), pp 86-92. https://doi.org/https://doi.org/10.2022/ajest.v3i3.57
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