Antifungal activity of Moringa oleifera oil and seed extract against ...

242 Middle East Journal of Agriculture Research, 3(2): 242-249, 2014 ISSN 2077-4605

Antifungal activity of Moringa oleifera oil and seed extract against some plant pathogenic fungi. 1

Riad S.R. El–Mohamedy and 2Aboelfetoh M. Abdallah

1 2

Plant Pathology Department, National Research Center, Cairo, Egypt. Horticulture Technology Department. National Research Center, Cairo, Egypt.

ABSTRACT The antifungal activities of Moringa oleifera oil and seed extract were investigated in vitro against seven plant pathogenic fungi i.e., Fusarium oxysporum, Fusarium solani, Alternaria solani, Alternaria alternate, Rhizoctonia solani , Sclerotium rolfsii and Macrophomina phaseolina . Results clearly showed that seed extracts and oil of Moringa oleifera were significantly reduced linear growth, spore germination and dry growth weight of all tested pathogens. M . oleifera oil and seed extract had different degrees of inhibition against growth rate and spore germination all tested pathogens. Reduction of growth and spore germination increased by increasing oil and seed extract. The highest records of reduction on both growth and spore germination were recorded for moringa oil at a concentration of 2.0%, followed by seed extract. F. oxysporum, F. solani and A. solani, A. alternate were highly affected by M. oleifera oil and seed extract than R. solani, S. rolfsii and M. phaseolina. Moringa oil and seed extract may be recommended as a potent bio-fungicide. Extensive studies should be undertaken for the Moringa oleifera extracts as a strong antifungal agent against fungal plant diseases under field conditions. Key words: Moringa oleifera , phytopathogenic fungi , Antifungal activity , spore germination . Introdction In recent years, interests have been generated in the development of safer antifungal agents from natural plant products such as, essential oils and extracts to control fungal diseases. Various plant materials are believed to have antifungal activity, as many essential oils and plant extracts have been reported to have antifungal activities with no side effects on humans and animals (Tabassum and Vidyasagar, 2013). Previous in vitro and in vivo investigations suggested that the essential oils and plant extracts could be used as effective antifungal agents against many phytopathogenic fungi (Sun et al., 2007; Lee et al., 2007; El-Mohamedy et al., 2013). Moringa (Moringa oleifera Lam.) has gained much importance in the recent days due to its multiple used and benefits to agriculture and industry. Regarded as a miracle plant, all parts of moringa plant are used for medicinal and other purposes (Price, 2000; Anwar et al., 2007). Recently, the roles of aqueous extracts of various plant parts in enhancing plant growth and productivity have been explored, making it even more valuable plant species (Anwar and Rashid, 2007). Investigations were carried out to evaluate the therapeutic properties of the seeds and leaves of Moringa oleifera Lam. as natural fungicides and herbal medicines (Bowers and Locke, 2000; Chuang et al., 2007; Satish et al., 2007; Dwivedi and Enespa, 2012; Tabassum and Vidyasagar, 2013) Fungal infections cause significant loss in many economic crops. Crop losses are estimated to be about 14% worldwide (Agrios 2005). Chemical control may be available to effectively and extensively reduce the effects of most fungal diseases but field application of these chemical fungicides may not always be desirable. Excessive and improper use of these fungicides presents a danger to the health of humans, animals, and the environment. Therefore, extensive searches for bio fungicides that are environmentally safe and easily biodegradable have been carried out during the last two decades (Gnanamanickam 2002). The investigation of plants containing natural antimicrobial metabolites for plant protection has been identified as a desirable method of disease control (Rai and Carpinella 2006). Various plant products like plant extracts, essential oils, gums, resins etc. were shown to exert biological activity in vitro and in vivo and are used as bio-fungicidal compounds (Fawzi et al. 2009; Al-Askar and Rashad 2010). The main reasons for using essential oils as antifungal agents are because they are natural in origin and because there is a low chance of pathogens developing resistance to them. Essential oils may have a minimum adverse effect on the physiological processes of plants and have less environmental hazards compared to their synthetic alternatives. Since essential oils are plant products, they are easily convertible into a common organic material and eco-friendly (Gnanamanickam 2002). Sseed extracts of Moringa oleifera were assayed for the evaluation of antimicrobial activity against bacterial (Pasturella multocida, Escherichia coli, Bacillus subtilis and Staphlocuccus aureus) and fungal (Fusarium solani and

Corresponding Author: Riad S.R. El–Mohamedy, Plant Pathology Department, National Research Center, Cairo, Egypt. E-mail: [email protected]

243 Middle East j. Agric. Res., 3(2): 242-249, 2014

Rhizopus solani) strains. The zones of growth inhibition showed greater sensitivity against the bacterial strains as compared to the fungal strains (Jamil et al. 2008). The aim of this work was to investigate the antifungal activity of seed extract and oil of Moringia olelfera in vitro, on the growth of seven phytopathogenic fungi i.e., Fusarium oxysporum , Fusarium solani, Alternaria solani, Alternaria alternate, Rhizoctonia solani, Sclerotium rolfsii and Macrophomina phaseolina the causal agents of many root rot, wilt and foliar diseases of many crops. Materials And Methods Plant materials: This study was carried out at the Department of Plant Pathology, National Research Center, Egypt. The source of Moringa oleifera oil and seeds kindly were obtained from Egyptian Scientific Society of Moringa (ESSM), National Research Center, Dokki, Cairo, Egypt. Fungal cultures: Seven plant pathogenic fungi such as Fusarium oxysporum, Fusarium solani, Alternaria solani, Alternaria alternate, Rhizoctonia solani, Sclerotium rolfsii and Macrophomina phaseolina were maintained and grown on potato dextrose agar medium. These pathogens were isolated and identified at Plant Pathology Department .National Resrach Center ,Cairo ,Egypt , the pathogenicty of each fungi were tested and recorded in previous studies by the author(El-Mohamedy et al., 2013). Preparation of morniga seed extract: Good quality dried Moringa oleifera seeds were selected and wings and coat from seeds were removed. Fine powder was prepared by using mortar and pestle and this powder was directly used aqueous extraction according to the method adopted by Price (2000). The aqueous extract prepared by this way in extraction; an amount of 100g of M. oleifera crashed seeds was boiled in 500 ml distilled water in a water path at 70 °C for 15 minutes and filtered. The filtrate was put in the freezer till freezing point, then extracted successfully by using Freeze drier apparatus for 96 hours. The supernatant layer was collected carefully in a clean container. Different concentrations (5, 10,15, 20, 25 %) of moringa seed extracts were prepared. Antifungal assay: Effect of moringa seed extract and oil on mycelia growth: Antifungal activity of moringa oil and/or seed extract was performed by the agar medium assay. Potato dextrose agar (PDA) medium with different concentrations of moringa seed extract (5, 10,15, 20, 25 %) as well as different concentrations of moringa oil (0.5, 1.0, 1.5, 2.0, 2.5 %) were prepared by adding appropriate quantity of moringa oil to melted medium, followed by addition of Tween 80 (100 µL to 100 mL of medium) to disperse the oil in the medium . About 20 ml of the medium were poured into glass Petri-dishes (9 cm x 1.5 cm). A 6 mm diameter agar disk bearing hyphae of either Fusarium oxysporum, Fusarium solani, Alternaria solani, Alternaria alternate, Rhizoctonia solani, Sclerotium rolfsii or Macrophomina phaseolinafrom 7-daysold colonies grown on PDA medium was transferred at the centre of each Petri-dish. Positive control (without moringa oil and or seed extract ) plates were inoculated following the same procedure. Plates were incubated at 25°C for 8 days and the colony diameter was recorded each day. Minimal inhibitory concentration (MIC) was defined as the lowest concentration of moringa oil in which no growth occurred. The MGI (Mycelia Growth Inhibition) percentage was calculated and expressed as percentage of reduction. Effect of moringa seed extract and oil on mycelia dry mass: The moringa oil and/or seed extract were mixed aseptically with sterilized Potato broth medium to produce concentrations 0.5, 1.0,1.5, 2.0, 2.5 % and 5, 10, 15, 20, 25 % of oil and seed extract respectively and dispensed in 50 mL aliquots into 250 mL Erlenmeyer flask. A 6 mm diameter agar disk bearing hyphae of either F. oxysporum, F. solani, A. solani, A. alternate, R. solani, S. rolfsii or M. phaseolinafrom 7-days-old colonies grown on PDA medium was transferred to each flask and incubated at 27±1°C for 10 days. Five flasks were prepared for each treatment .The mycelia were harvested, dried to constant weight at 80±l°C, the dry mass yield was recorded and the percentage of reduction in mass production was calculated


Effect of moringa seed extract and oil on spore and sclertia germination: Sclerotia of R. solani, S. rolfsii and M . phaseolina produced on potato dextrose agar (PDA) were collected and surface disinfected by soaking them for 5 min in 1:400 (w/v) bromine in water to kill hyphal extension, washed thoroughly with distilled water and dried . Ten sclerotia/Petri dish for either pathogen were plated on the surface of tap water agar (1.5% w/v) supplemented with different concentrations of oil and/or seed extract of moringa maintained above. The dishes were incubated at 27±1°C for 24 h the percentage of germinated sclerotia were determined and five plates were prepared for each treatment. For F. oxysporum, F. solani, A. solani and A. alternate, microscope slides were covered, each, with 1 mL of spores suspension of each pathogen in aqueous solution of the desired oil and/or seed extract concentrations in Petri dishes and then incubated at 27±1°C for 8 h in complete darkness. The percentage of germination was assessed according . Five plates were prepared for each treatment and the means were compared. Statistical analyses: All experiments were set up in a complete randomized design. One-way ANOVA was used to analyze differences between antagonistic inhibitor effect and mycelial growth of pathogenic fungi in vitro. A general linear model option of the analysis system SAS (48) was used to perform the ANOVA. Duncan’s multiple range tests at P < 0.05 level was used for means separation (Winer, 1971). Results: Fungicidal activity against myceliae linear growth, spore /sclerotia germination and dry growth amount weight of seven pathogenic plant fungi i.e., Fusarium oxysporum, Fusarium solani, Alternaria solani, Alternaria alternate, Rhizoctonia solani, Sclerotium rolfsii and Macrophomina phaseolina was observed at different concentrations of M. oleifera oil and seed extracts in vitro. 1-Antifugal activities of Moringa oleifera oil: Efficiency of M. oleifera oil at different concentrations in decreasing linear growth, spore / sclerotia germination and growth amount dry weight of the investigated pathogenic plant fungi was studied. Results in Table (1) show that fungal mycelial growth was sharply decreased with increasing concentration of moringa oil to reach minimum growth at 2.0%. Moringa oil at 2.5% completely inhibit the linear growth of all tested pathogens. F. oxysporum and F. solani were complete hampered at 2.0% of moringa oil, but the growth of A. solani, A. alternate inhibited by 86.6 and 75.6 % .Meanwhile, R. solani, S. rolfsii and M. phaseolina showed growth reduction reach to 81.1, 74.8, and 79.4 % respectively at 2.0% moringa oil concentration. Mycelial growth of F. oxysporum, F. solani, A. solani, A. alternate showed more sensitivity to morigna oil than R. solani , S. rolfsii and M. phaseolina. It is clear to state that moringa oil at all tested concentration had antifungal activity against the most root rot and foliar disease pathogens under this investigation. Table 1: Effect of M. oleifera oil at different concentrations on Linear growth (mm) of some pathogenic plant fungi on PDA medium . M. oleifera oil Concentration % Pathogenic fungi 0.0 % 0.5% 1.0% 1.5% 2.0% 2.5% D I D I D I D I D I D I Fusarium oxysporum 90a 0.0 38b 55.8 28c 68.8 16d 75.6 0.0e 100 0e 100 Fusarium solani 90a 0.0 40b 54.0 26c 66.6 12d 70.2 0.0e 100 0e 100 Alternaria solani 90a 0.0 56b 39.6 42c 53.3 31d 62.1 12e 86.6 0e 100 Alternaria alternate 90a 0.0 60b 36.0 45c 50.0 36d 57.6 16e 75.6 0f 100 Rhizoctonia solani 90a 0.0 68b 28.8 52c 43.2 40d 54.0 17e 81.1 0f 100 Sclerotium rolfsii 90a 0.0 70b 27.0 60c 36.0 43d 51.3 22e 74.8 0f 100 Macrophomina phaseolina 90a 0.0 65b 27.7 57c 38.7 46d 58.6 18e 79.4 0f 100 D= colony diameter (mm) I = Inhibition percentage (%) as compared to the control (0.0 %) . Means followed by the same letters are not significantly different according to Duncan’s multiple range test (P ≤ 0.05).

Studies on spore germination represent and integral part of the ecological studies of the pathogenic fungi, as spores are the specialized structures capable of initiating new growth. Once germination had occurred, the ensuring mycelia growth rate may be of prime importance in determining the degree of virulence of the fungus concerned. The antifungal efficiency of moringa oil towards spore germinationand dry growth weight of seven phytopathogenic fungi was studied in vitro and the results are presented in Figar (1). Figar (1) show cleary that all concentrations of moringa oil inhibit spore germination of F. oxysporum, A. solani and A. alternate as well as sclertia germination of R. solani , S. rolfsii and M. phaseolina with different


values. Moringa oil at 2.5 % completely inhibit spores and sclerotia germination of all tested pathogens. Moringa oil at 2.0 % reduced spores germination of F. oxysporum and F. solani by 100%; A. solani, A. and A. alternate by 92.2 and 90.0 %, sclerotia germination of R. solani, S. rolfsii and M. phaseolina by 84.4 , 82.2 and 95.8 % respectively. Spores germination of F.oxysporum , F. solani , A. solani and A. alternate showed more sensitivity to morigna oil than R. solani, S. rolfsii and M. phaseolina. Concerning the effect of moringa oil on dry growth amount weight of all tested pathogens, the same trend of result was observed also in Figure(1). Moringa oil at 2.5 % was the most effective in decreasing dry growth amount of F. oxysporum , F. solani, A. solani, A. alternate, R. solani, S. rolfsii and M. phaseolina (100%). At 1.5 and 2.0 % concentrations of moringa oil reduction varies from 51.2% to 88.6 % and 71.4 % to 96.0% of moringa oil. In the lowest concentrations 0.5 and 1.0 % of moringa oil minimum inhibition of dry growth amount as recoded by R solani, S rolfsii and M. phaseolina .The most affected pathogens by all concentrations of moringa oil were F. oxysporum , F. solani followed by A. solani and A. alternate, but R. solani, S. rolfsii and M. phaseolina showed less sensitivity against moringa oil compeered with other pathogens. Moringa oil show high effectiveness against spore and sclerotia germination of all tested pathogens than on dry growth amount of these pathogens.

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Fig. 1: Reduction (%) of spore/sclerotia germination (SG) and dry growth (DW) amount of some phtyopathogenic fungi in response to different concentrations of Moringa oleifera oil in vitro


2-Antifungal activities of Moringia oleifera seed extract (MSE): Different concentrations (5,10,15, 20, 25 %) of moringa oliefera seeds extracts were tested against F. oxysporum, F. solani, A. solani, A. alternate, R. solani, S. rolfsii and M. phaseolina to determine their antifungal activity in vitro tests. Results in Table (2) Cleary show that fungicidal activity against linear growth of all tested pathogens was observed at all concentrations of moringa seed extract (MSE) .The inhibitory effect against all tested pathogens increased by increasing MSE concentration to reach maximum effect (complete reduction) at 25 % of MSE . Moringa seed extract at 20 % completely reduce (100%) the linear growth of F. oxysporum , F. solani , A.solani , A. alternate , and by 94.4 , 81.9 , 92.2 % of R. solani , S. rolfsii and M. phaseolina .At 10 % and 20 % concentrations of MSE, growth reduction varies from 66.7 % to 82.2 % and 82.2 % to 95.5 % of all tested pathogens respectively . Mycelial growth of F. oxysporum , F. solani , A. solani , A. alternate showed more sensitivity to morigna seed extract (MSE) than R. solani , S. rolfsii and M. phaseolina. It is clear to state MSE had antifungal activity against the most roots rot and foliar plant pathogens under this investigation. Table 2: Efficiency of M. oleifera seed extract at different concentrations on Linear growth (mm) of some pathogenic plant fungi on PDA medium. M. oleifera seed extract concentration % Pathogenic fungi 0.0 % 5% 10% 15% 20% 25% D I D I D I D I D I D I Fusarium oxysporum 90a 0.0 30b 66.7 19b 82.2 4c 95.5 0c 100 0c 100 Fusarium solani 90a 0.0 30b 66.7 18b 80.0 7c 92.2 0c 100 0c 100 Alternaria solani 90a 0.0 60b 33.3 18b 80.0 8c 91.1 0d 100 0d 100 Alternaria alternate 90a 0.0 72b 20.0 13b 85.5 8c 91.1 0d 100 0d 100 Rhizoctonia solani 90a 0.0 54b 41.0 28b 68.9 11c 87.8 5d 94.4 0d 100 Sclerotium rolfsii 90a 0.0 64b 32.0 36b 60.0 18c 80.0 9d 81.9 0c 100 Macrophomina phaseolina 90a 0.0 65b 30.0 30b 66.7 16c 82.2 7d 92.2 0c 100 D= colony diameter (mm) I = Inhibition percentage (%) as compared to the control (0.0 %) . Means followed by the same letters are not significantly different according to Duncan’s multiple range test (P ≤ 0.05).

M. oleifera seed extract cause decreasing in percentage of spore /sclerotia germination as well as amount of dry growth of all tested pathogens (Figure 2). Results in Figure(2) demonstrated that all concentrations of moringa seed extract (MSE) hampered spore germination of F. oxysporum, F. solani, A. solani and A. alternate as well as sclertia germination of R. solani, S. rolfsii and M. phaseolina with different values. Moringa seed extract at 20.0 and 25 % cause complete (100%) inhibition of germinated spores and/or sclerotic of all tested pathogens. Moringa seed extract at 15 % reduced spores germination of F. oxysporum and F. solani by 100%; A. solani and A. alternate by 88.8 and 86.2 % . Meanwhile, it reduced sclerotia germination of R. solani, S. rolfsii and M. phaseolina by 90.0, 85.4 and 92.2 % respectively. At lowest concentrations of MSE (5 % and 10%) reduction of germinated spores and/or sclerotia varies from 40.6 % to 65.2 % and 71.2 % to 82.0 % of all tested pathogens respectively . Spores germination of F. oxysporum, F. solani, A. solani and A. alternate showed more sensitivity to morigna seed extract than R. solani, S. rolfsii and M. phaseolina. Concerning the effect of moringa seed extract on dry growth amount weight of all tested pathogens, the same trend of result was observed in Figure(2). Concentrations of Moringa seed extract at 20 and 25 % were the most effective in decreasing dry growth weight of F. oxysporum, F. solani, A. solani, A. alternate R. solani, S. rolfsii and M. phaseolina (100%). Reduction in dry growth of all tested pathogens was varies from 63.6 % to 82.0 % and 85.4 % to 100 % at 10 and 15 % concentrations of moringa seed extract. In the lowest concentrations 5 % of moringa seed extract , minimum inhibition of dry growth amount as recoded by R. solani, S. rolfsii and M. phaseolina. Moringa seed extract at all concentrations show high effectiveness against spore and sclerotia germination of all tested pathogens than on dry growth amount of these pathogens. The most affected pathogens by all concentrations of moringa seed extract were F. oxysporum, F. solani followed by A. solani and A. alternate, but R. solani, S. rolfsii and M. phaseolina showed less sensitivity against moringa oil compeered with other pathogens


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Fig. 2: Reduction (%) of spore/sclerotia germination (SG) and dry growth (DW) amount of some phtyopathogenic fungi in response to different concentrations of Moringa oleifera seed extract in vitro. Discussion: Synthetic fungicides are currently used as the primary means for the control of plant diseases. However, the alternative control methods are needed because of the negative public perceptions about the use of synthetic chemicals, resistance to fungicides among fungal pathogens, and high development cost of new chemicals. The uses of plant derived products as diseases control agents have been studied, since they tend to have low mammalian toxicity, less environmental effects and wide public acceptance (Seema et al., 2011; Moyo et al., 2012). To develop environment-friendly alternatives to synthetic fungicides for the control of fungal plant diseases, the interest on essential oils and plant extracts has been increased. (Gnanamanickam 2002) In this study, we investigated the antifungal activities of Moringa oleifera oil and seed extract against seven plant pathogenic fungi i.e., F. oxysporum, F. solani, A. solani, A. alternate, R. solani, S. rolfsii and M. phaseolina. Our results demonstrated that Moringa oleifera oil and seed extract at all tested concentrations had considerable effect on the growth rate and spore germination of all tested pathogens. Moringa oil at 2.5% completely inhibit the linear growth of all tested pathogens. F. oxysporum and F. solani were complete


hampered at 2.0% of moringa oil ,but the growth of A. solani, A. alternate inhibited by 86.6 and 75.6 % .Meanwhile, R. solani, S. rolfsii and M. phaseolina showed growth reduction reach to 81.1, 74.8, and 79.4 % respectively . The most affected pathogens by all concentrations of moringa oil were F. oxysporum, F. solani followed by A. solani and A. alternate, but R. solani, S. rolfsii and M. phaseolina showed less sensitivity against moringa oil compeered with other pathogens . Similar studies have been carried out by different researcher on antifungal activity of essential oil and extract of many plants (Satish et al., 2007 ; Jamil et al., 2007 ; Anwar and Rashid, 2007;Sun et al., 2007). The essential oil of Thymus vulgaris suppressed the mycelial growth of C. gloeosporioides, Fusarium oxysporum and Rhizoctonia solani and that of Cymbopogon citratus was active to only F. oxysporum (Lee et al., 2007). Many essential oils and plant extracts have been found to be potent fungitoxic agents against many plant pathogens (Siripornvisal and Ngamchawee, 2010; El-Mohamedy et al., 2013; Tabassum and Vidyasagar, 2013; Abd el-kader et al., 2013). However, the harmful effects on fungi were restricted in: (a) partial or complete inhibition on spore germination, sporulation or mycelia growth and (b) alternation in physiology and biochemistry activities of the fungal cells. Several in vitro studies have been published confirming the effect of essential oil and their major compounds on plant and human pathogenic fungi (Lee et al., 2007; Chuang et al., 2007; Tabassum and Vidyasagar, 2013; Hadi and Kashefi, 2013). Studies on spore germination represent and integral part of the ecological studies of the pathogenic fungi, as spores are the specialized structures capable of initiating new growth. Our studies also show that Moringa seed extract at 20 % completely reduce (100%) the linear growth of F. oxysporum, F. solani, A. solani, A. alternate, and by 94.4, 81.9, 92.2 % of R. solani, S. rolfsii and M. phaseolina. In addition, Moringa seed extract at 20.0 and 25 % cause complete (100%) inhibition of germinated spores and/or sclerotia of all tested pathogens. Spores germination of F. oxysporum, F. solani, A.solani, A. alternate and M. phaseolina showed more sensitivity to morigna oil than R. solani and S. rolfsii. Moringa seed extract at 20 and 25 % were the most effective in decreasing dry growth weight of F. oxysporum , F. solani, A. solani, A. alternate R. solani and S. rolfsii and M. phaseolina (100%). These finding are in agreement with these reported by many investigators (Bowers and Locke, 2000 ; Dwivedi and Enespa, 2012; Abdulmoneim et al., 2011; Abandonon et al., 2006; Chuang et al., 2007) In this respect, Jamil et al. (2008) evaluated antimicrobial activity of Moringa oleifera against bacterial (Pasturella multocida, Escherichia coli, Bacillus subtilis and Staphlocuccus aureus) and fungal (Fusarium solani and Rhizopus solani) strains. The zones of growth inhibition showed greater sensitivity against the bacterial strains as compared to the fungal strains. Reports have been elucidated on the findings of the antibiotic principle of M. oleifera seeds through their purification, elucidation, and antimicrobial properties, and also on the antibiotic substance of the roots of M. oleifera (Jamil et al., 2008; Bowers and Locke, 2000; Dwivedi and Enespa, 2012). The findings in this study confirmed that Moringa oleifera oil and seed extract can be used as natural fungicides to control pathogenic fungi and thus reduce the dependence on the synthetic fungicides Conclusion: Chemicals from plants, which retards the reproduction of undesirable microorganisms, would be a more realistic and ecologically sound method for plant protection and will have a prominent role in the development of future commercial pesticides for crop protection strategies with special reference to the management of plant diseases. The results obtained in the present work indicate that Moringa oleifera oil and seed extract had antifungal activity against seven plant pathogenic fungi tested in the present study in vito. So, it is interested to state that we can used Moringa oleifera oil and/or extracts as eco-friendly means, environmentally safe, less risky for developing resistance in pests, and pest resurgence, has less adverse effect on plant growth, less harmful to seed viability and quality, and above all, less expensive for controlling plant diseases under field. Referencese Abdel-Kader, M.M., F. Abdel-Kareem, N.S. El-Mougy and R.S. El-Mohamady, 2013. Integration between Compost, Trichoderma harzianum and Essential Oils for Controlling Peanut Crown Rot under Field Conditions Journal of Mycology, 7: 1-7. Abdulmoneim, M., M. Saadabi and I.E. Abu Zaid, 2011. An In vitro Antimicrobial Activity of Moringa oleifera L. Seed Extracts Against Different Groups of Microorganisms. Australian Journal of Basic and Applied Sciences, 5(5): 129-134, 2011. Adandonon, T., A.S. Aveling, N. Labuschagne and M. Tamo, 2006. Biocontrol agents in combination with Moringa oleifera extract for integrated control of Sclerotium-caused cowpea damping-off and stem rot. European Journal of Plant Pathology. Agrios, G.M., 2005. Plant Pathology. 5th ed. AP, New York, NY, pp: 922.


Al-Askar, A.A. and Y.M. Rashad, 2010. Efficacy of some plant extracts against Rhizoctonia solani on pea. J. Plant Prot. Res., 50(3): 239-243. Anwar, F. and U. Rashid, 2007. Physico-chemical characteristics of Moringa Oleifera seeds and seed oil from a wild provenance of Pakistan. Pak. J. Bot., 39(5): 1443-1453. Anwar, F., S. Latif, M. Ashraf and A.H. Gilan, 2007. Moringa oleifera: A Food plant with Multiple Medicianl uses, Phytotherapy Research, 21: 17-25. Bowers, J.H. and J.C. Locke, 2000. Effect of botanical extracts on the population density of Fusarium oxysporum in soil and control of Fusarium wilt in the greenhouse, Plant Disease, 84: 300-305. Chuang, P.H., C.W. Lee, J.Y. Chou, M. Murugan, B.J. Shieh and H.M. Chen, 2007. Antifungal activity of crude extracts and essential oil of Moringa oleifera Lam. Biores Tech., 98: 232-236. Dwivedi, S.K. and A. Enespa, 2012. Effectiveness of extract of some medical plants against soil borne fusaria causing diseases on Lycopersicon esculantum and Solanum melongena. Int. J. Pharm. Bio. Sci., 3(4): 11711180. El-Mohamedy, R.S.R., M.M. Abdel-Kader, F. Abd-El-Kareem and N.S. El-Mougy, 2013. Essential oils, inorganic acids and potassium salts as control measures against the growth of tomato root rot pathogens in vitro. Journal of Agricultural Technology, 9(6): 1507-1520. Hadi, M. and B. Kashefi, 2013. Study on Effect of Some Medicinal Plant Extracts on Growth and Spore Germination of Fusarium oxysporum schlecht. In vitro American-Eurasian .J. Agric. and Environ. Sci., 13(4): 581-588. Fawzi, E.M., A.A. Khalil, A.F. Afifi, 2009. Antifungal effect of some plant extracts on Alternaria alternata and Fusarium oxysporum. Afr. J. Biotechnol., 8(11): 2590-2597. Jamil, A., M. Shahid, M.M. Khan and M. Ashraf, 2007. Screening of some medicinal plants for isolation of antifungal proteins and peptides. Pak. J. Bot., 39(1): 211-221. Gnanamanickam, S.S., 2002. Biological Control of Crop Diseases. New York. Basel: Marcel Dekker, Inc., pp: 15. Lee, S.O., J.C. Gyung, S.J. Kyoung, L. He Kyoung, Y.C. Kwang and K. Jin-Cheol, 2007. Antifungal Activity of Five Plant Essential Oils as Fumigant Against Postharvest and Soilborne Plant Pathogenic Fungi. Plant Pathol. J. 23(2): 97-102 Moyo, B., P.J. Masika and V. Muchenje, 2012. Antimicrobial activities of Moringa oleifera Lam leaf extracts, African Journal of Biotechnology, 11(11): 2797-2802. Rai, M. and M. Carpinella, 2006. Naturally Occurring Bioactive Compounds. Elsevier, Amsterdam, p. 502. Price, M.L., 2000. The Moringa Tree purification and activity assay of the coagulant protein from Moringa oleifera seed. Water Res., 39: 2338-2344. Siripornvisal, S. and K. Ngamchawee, 2010. Utilization of herbal essential oils as biofumigant against fungal decay of tomato during storage. Proceedings 16th Asian Agricultural Symposium, Bangkok, Thailand., pp: 655-658. Satish, S., D.C. Mohana, M.P. Raghavendra, K.A. Raveesha, 2007. Antifungal activity of some plant extracts against important seed borne pathogens of Aspergillus sp. J. of Technol., 3: 109-119. Seema, M.S.S., N.D. Sreenivas, Rekha and N.S. Devaki, 2011. In vitro studies of some plant extracts against Rhizoctonia solani Kuhn infecting FCV tobacco in Karnataka Light Soil, Karnataka, India. Journal of Agricultural Technology, 7(5): 1321-1329. Sun, O.L., J.C. Gyung, S.J. Kyoung, K.L. He, Y.C Kwang and K. Jin-Cheol, 2007. Antifungal Activity of Five Plant Essential Oils as Fumigant against Postharvest and Soil borne Plant Pathogenic Fungi. Plant Path J. 23(2): 97-102. Tabassum, N. and G.M. Vidyasagar, 2013. Antifungal investigations on plant essential oils, A review. Int J Pharm Pharm Sci., 5(2): 19-28. Winer, B.J., 1971. Statistical Principles in Experimental Design. Page 596. In 2nd ed. MiGraw-Hil Kogakusha, LTD.