ABSTRACT Fungi of the genus Fusarium are well known plant pathogens, cause several vascular diseases and are producers of toxins. In vitro assays evaluated the effects of Neem (Azadirachta indica) oil on the diameter of colonies, dry weight, spore production, spore viability and production of Fusaric Acid toxin on Fusarium oxysporum f. sp. medicagenis and Fusarium subglutinans isolates. Effects of Neem oil were analyzed at concentrations 0.25%, 0.5% and 1% in Czapek Yeast Agar medium. The production of Fu- saric acid was determined by Thin Layer Chromatography and quantified by UV spectrophotometry. Neem oil showed inhibitory effects on the isolates tested, although they varied according to type of isolate and oil concentration. Neem oil was efficient in reducing the colonies’ diameter and dry weight in concentration-dependent manner. Neem oil was efficacious at higher concentration in the decrease of sporulation. Spore germination was affected by Neem oil when the spore was grown in Neem-contained medium as when the spore emerged from a culture in a Neem medium. Neem oil decreased and even inhibited the production of Fusaric acid by the assayed isolates. Since these isolates are plant pathogens and producers of Fusaric acid, Neem oil may be introduced as an integral item in the management of host plants.
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nullGeraldo, M. , Arroteia, C. and Kemmelmeier, C. (2010) The effects of neem [Azadirachta indica A. Juss (meliaceae)] oil on Fusarium oxysporum f. sp. medicagenis and Fusarium subglutinans and the production of fusaric acid toxin. Advances in Bioscience and Biotechnology, 1, 1-6. doi: 10.4236/abb.2010.11001.
 Pitt, J.I. and Hocking, A.D. (1997) Fungi and food spoilage. 2nd Edition, Blackie Academic and Professional, London.
Velutti, A., Marin, S., Gonzalez, P., Ramos, A.J. and Sanchis, V. ( 2004) Initial screening for inhibitory activity of essential oils on growth of Fusarium verticillioides, F. proliferatum and F. graminearum on maize-based agar media. Food Microbiology, 21, 649-656.
Booth, C. (1971) The genus fusarium. Commonwealth Agricultural Bureaux, England.
Seifert, K.A., et al. (2003) The name Fusarium moniliforme should no longer be used. Mycological Research, 107, 641-642.
Porter, J.K., Bacon, C.W., Wray, E.M. and Hagler, W.M.J. (1995) Fusaric acid in Fusarium moniliforme cultures, corn, and feeds toxic and the neurochemical effects in the brain and pineal glands of rats. Natural Toxins, 3, 91-100.
Hidaka, H. and Nagatsu, T. (1969) Fusaric acid, a hypotensive agent produced by fungi. Journal of Antibiotics, 22, 228-230.
Nagatsu, T., Hidaka, H., Kuzuya, H., Takeya, K., Umezawa, H., Takeuchi, T. and Suda, H. (1970) Inhibition of dopamine β-hydroxylase by fusaric acid (5-butylpicolinic acid) in vitro and in vivo. Biochemical Pharmacology, 19, 35-44.
Bacon, C.W., Porter, J.K. and Norred, W.P. (1995) Toxic interaction of fumonisin B1 and fusaric acid measured by injection into fertile chicken egg. Mycopathology, 129, 29-35.
Bacon, C.W., Porter, J.K., Norred, W.P. and Leslie, F. Production of fusaric acid by Fusarium species. Applied and Environmental Microbiology, 62, 4039-4043.
Luz, J.M., Paterson, R.R.M. and Brayford, D. (1990) Fusaric acid and other metabolite production in Fusarium oxysporum f. sp. Vasinfectum. Letters in Applied Microbiology, 11, 141-144.
Marrè, M.T., Vergani, P. and Albergoni, F.G. (1993) Relationship between fusaric acid uptake and its binding to cell structures by leaves of Egeria densa and its toxic effects on membrane permeability and respiration. Physiological and Molecular Plant Pathology, 42, 141-157.
Samad, L. and Behboodi, B.S. (1985) Fusaric acid induces apoptosis in savron root tip cells: roles of caspase-like activity, cytochrome c and H2O2. Planta, 225, 213-234.
Arias, J.A. (1985) Secretory organelle and mitochondrial alterations induced by fusaric acid in root cells of Zea mays. Physiological and Plant Pathology, 27, 149-158.
Bouizgarne, B., Brault, M., Pennarun, A.M., Rona, J.P., Ouhdouch, Y., El Hadrami, I. and Bouteau, F. (2004) Electrophysiological responses to fusaric acid of root hairs from seedlings of date palm-susceptible and -resis- tant to Fusarium oxysporum f. sp. Albedinis. Journal of Phytopathology, 152, 321-324.
Curir, P., Guglieri, L., Dolci, M., Capponi, A. and Aurino, G. (2000) Fusaric acid production by Fusarium oxysporum f.sp. lilii and its role in the lily basal rot disease. European Journal of Plant Pathology, 106, 849-856.
Wu, H.S., Yin, X.M., Zhu, Y.Y., Guo, S.W., Wu, C.L., Lu, Y.L. and Shen, Q.R. (2007) Nitrogen metabolism disorder in watermelon leaf caused by fusaric acid. Physiological and Molecular Plant Pathology, 71, 69-77.
Kuzniak, E. (2001) Effects of fusaric acid on reactive oxygen species and antioxidants in tomato cell cultures. Journal of Phytopathology, 149, 575-582.
Capasso, R., Evidente, A., Cutignano, A., Vurro, M., Zonno, M.C. and Bottalico, A. (1996) Fusaric and 9,10- dehydrofusaric acids and their methyl esters from Fusarium nygamai. Phytochemistry, 41, 1035-1039.
Barna, B. and Györgyi, B. (1992) Resistance of young versus old tobacco leaves to necrotrophs, fusaric acid, cell wall-degrading enzymes and autolysis of membrane lipids. Physiological and Molecular Plant Pathology, 40, 247-257.
Gapillout, L., Milat, M.L. and Blein, J.P. (1996) Effects of fusaric acid on cells from tomato cultivars resistant or susceptible to Fusarium oxysporum f. sp. Lycopersici. European Journal of Plant Pathology, 102, 127-132.
Chao, S.C., Young, D.G. and Oberg, C.J. (2000) Screening for inhibitory activity of essential oils on selected bacteria, fungi and viruses. Journal of Essential Oil Research, 12, 639-649.
Cakir, A., Kordali, S., Kilic, H. and Kaya, E. (2005) Antifungal properties of essential oil and crude extracts of Hypericum linarioides Bosse. Biochemical Systematics and Ecology, 33, 245-256.
Mossini, S.A.G. and Kemmelmeier, C. (2005) A árvore Nim (Azadirachta indica A. Juss): Múltiplos Usos. Acta Farmaceutica Bonaerense, 24, 139-148.
Neves, B.P., Oliveira, I.P. and Nogueira, J.C.M. (2003) Cultivo e utilização do nim indiano. Embrapa Circular Técnica, 62, Goiás.
Locke, J.C. (1995) Fungi, H., Ed. In Schmutterer, The Neem Tree. VHC, Weiheim, Germany, 118-126.
Martinez, S.S. (2002) O Nim – Azadirachta indica. Natureza, Usos Múltiplos, Produção. Martinez, S.S., Ed., IAPAR, Londrina.
Bowers, J.H. and Locke, J.C. (2004) Effect of formulated plant extracts and oils on population density of Phytophthora nicotianae in soil and control of phytophthora blight in the greenhouse. Plant Disease, 88, 11-16.
Hirose, E., Neves, P.M.O.J., Zequi, J.A.C., Martins, L.H., Peralta, C.H. and Moino, A.J. (2001) Effect of biofertilizers and neem oil on the entomopathogenic fungi Beauveria bassiana (bals.) vuill and Metarhizium anisopliae (metsch.) sorok. Brazilian Archives of Biology and Technology, 44, 419-423.
Krishnamurthy, Y.L. and Shashikala, J. (2006) Inhibition of aflatoxin B1 production of Aspergillus flavus, isolated from soybean seeds by certain natural plant products. Letters in Applied Microbiology, 43, 469-474.
Razzaghi-Abyaneh, M., Allameh, A., Tiraihi, T., Ghahfarokhi, M.S. and Ghorbanian, M. (2005) Morphological alteration in toxigenic Aspergillus parasiticus exposed to neem (Azadirachta indica) leaf and seed aqueous extracts. Mycopathologia, 159, 565-570.
Zeringue, H.J.J. and Bhatnagar, D. (1994) Effects of neem leaf volatiles on submerged cultures of aflatoxigenic Aspergillus parasiticus. Applied and Environmental Microbiology, 60, 3543-3547.
Arrotéia, C.C., Kemmelmeier, C. and Machinski, J.M. (2007) Efeito dos extratos aquoso e oleoso de Nim [Azadirachta indica A. Juss (Meliaceae)] na produção de patulina em maçãs contaminadas por Penicillium exp- ansum. Ciência Rural, 37, 1518-1523.
Costa, C.L. and Kemmelmeier, C. (2008) Effect of aqueous and oily extracts from Neem [Azadirachta indica A. Juss (Meliaceae)] on the production of mycotoxins by the polyketide pathway (penicillic acid and sterigmatocystin). Current Topics of Biotechnology, 4, 35-40.
Mossini, S.A.G., De Oliveira, K.P. and Kemmelmeier, C. (2004) Inhibition of patulin production by Penicillium expansum cultured with Neem (Azadirachta indica) Leaf Extracts. Journal of Basic Microbiology, 44,106-113.
Mossini, S.A.G. and Kemmelmeier, C. (2008) Inhibition of citrinin production in Penicillium citrinum by Azad- irachta indica A. Juss (Meliaceae) in culture. International Journal of Molecular Sciences, 9, 1676-1684.
Mossini, S.A.G., Arrotéia, C.C. and Kemmelmeier, C. (2009) Effect of Neem leaf extract and Neem oil on Penicillium growth, sporulation, morphology and Ochratoxin A production. Toxins, 1, 2-13.
Marques, R.P., Monteiro, A.C. and Pereira, G.T. (2004) Crescimento, esporulação e viabilidade de fungos ento- mopatogênicos em meios contendo diferentes concen- trações do óleo de Nim (Azadirachta indica). Ciência Rural, 34, 1675-1680.
Paterson, R.R.M. and Rutheford, M.A. (1991) A simplified rapid technique for fusaric acid detection in Fusarium strains. Mycopathologia, 113, 171-173.
Eged, S. (2005) Thin-layer chromatography – an appropriate method for fusaric acid estimation. Biologia Bratislava, 60, 104.
Adams, T.H. and Yu, J-H. (1998) Coordinate control of secondary metabolite production and asexual sporulation in Aspergillus nidulans. Current Opinion in Microbiology, 1, 674-677.
Pawar, V.C. and Thaker, V.S. (2006) In vitro efficacy of 75 essential oils against Aspergillus niger. Mycoses, 49, 316-323.
Soliman, K.M. and Badeaa, R.I. (2002) Effect of oil extract from some medicinal plants on different mycotoxigenic fungi. Food and Chemical Toxicology, 40, 1669- 1675.
Fandohan, P., Gbenou, J.D., Gnonlonfin, B., Hell, K., Marasas, W.F.O. and Wingfield, M.J. (2004) Effect of essential oils on the growth of Fusarium verticillioides and fumonisin contamination in corn. Journal of Agricultural and Food Chemistry, 52, 6824-6829.
Agbenin, O.N. and Marley, P.S. (2006) In vitro assay of some plant extracts against Fusarium oxysporum f. sp. lycopersici causal agent of tomato wilt. Journal of Plant Protection Research, 46, 215-220.
Schaaf, O., Jarvis, A.P., van der Esch, S.A., Giagnacovo, G. and Oldham, N.J. (2000) Rapid and sensitive analysis of azadirachtin and relate triterpenoids from neem (Azadirachta indica) by high performance liquid chromatography - tmospheric pressure chemical ionization mass spectrometry. Journal of Chromatrgraphy A, 886, 89-97.