World Journal of Science and Technology 2012, 2(6):58-62 ISSN: 2231 2587 Available Online: www.worldjournalofscience.

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Antifungal potential of crude plant extract on some pathogenic fungi

*Jaya Gujar and Dhananjay Talwankar *Singhania University (Raj), Dept. of Botany G. S. Sci, Arts and Comm. College, Khamgaon, Dist- Buldana (MS), India. Abstract In the present work, the antifungal activity of plant extract of selected medicinal and locally available plant on some pathogenic fungi is tested. Six different plants used for the testing and these plant are Azadirachta indica, Aloe vera, Ocimum sanctum, Ocimum basilicum, Lantana camara and Asparagus. These plants showed the antifungal activity against the Aspergillus niger, Aspergillus flavus, Rhizoctonia solani, Rhizoctonia bataticola. These plants exhibited varying degrees of inhibition activity against the fungi. Antifungal activity was tested against the four pathogens attacking commercial crop. All selected plant exhibited considerable distinction in radial mycelial growth of tested pathogen. Over all, Azadirachta indica and Aloe vera appeared significantly the most effective and suppressed the radial mycelial growth of Aspergillus and Rhizoctonia species. Whereas Ocimum sanctum exhibited maximum inhibition (90-95%), against Aspergillus niger. However Ocimum basilicum and Lantana camara exhibited moderate type of inhibition against all tested pathogen and Asparagus shows least potential of inhibition against all tested pathogen. It was also observed that radial mycelial growth of selected pathogen reduced in alcoholic solvent. Among three solvent highest inhibitions in radial mycelial growth of all four pathogen was observed in alcohol. And acetone shows moderate inhibition while minimum inhibition was recorded in water extract of plant. It may be concluded from the present investigation that Azadirachta indica and Aloe vera can be utilized for the management of fungal disease caused by the Aspergillus niger, Aspergillus flavus, Rhizoctonia solani, Rhizoctonia bataticola. Keywords: Pathogenic Fungi,Antifungal, plant extract. INTRODUCTION Biological screening of plant extract is carried out throughout the world for the determination of their antifungal activity. Large number of synthetic chemicals and fungicides used to control plant disease not only pollute the environment but are also harmful to human health because of that environmental considerations, many workers are involved to find the cheaper and more environment ecofriendly bio compounds for the control of plant disease using diffusates from different plants (Gerresten and Haagsma 1951). The use of Neem cake and extract as a soil treatment measure have produced good result against various soil borne fungi like Pythium aphanidermatum and Rhizoctonia solani,(khan et al 1974), Fusarium oxysporum (kannaiyan and Prasad 1981), Colletotrichum atramentarium (sing1986) Effectiveness of Neem extract and oil as a fungicide has earlier been reported by several worker (R. C. Dubey et al 2009, Ilyas et al1997, Sharma and Basandrai1997, Lokhande et. Al 1998, Dubey and kumar 2003) found almost similar effect of Azadirachta on growth of M. phaseolina. Antifungal activity of selected medicinal plant diffusates against Alternaria solani, Rhizoctonia solani and M. phaseolina also Aspergillus niger causing crown rot of groundnut , the most serious plant disease . The methanolic extract of 43 plants exhibited
Received: April 12, 2012; Revised: May 15, 2012; Accepted: June 25, 2012. *Corresponding Author Jaya Gujar Singhania University (Raj), Dept. of Botany G. S. Sci, Arts and Comm. College, Khamgaon, Dist- Buldana (MS), India. Email: jayagujar@gmail.com

varying degree of inhibition activity against the fungi ( Vara Prasad Babbura la et al 2009) alcoholic extract of Neem used for retarding the growth of Aspergillus species (Monaali N.K. et al 2009). Rhizoctonia solani an important destructive soil borne pathogen has detrimental effects on agricultural and horticultural crops by pre-emergence and post emergence damping off , root rot, and stem canker. There are over five hundred hosts in United States of America alone (Farr et al, 1989). It can affect potato plants from planting to harvest by inhibition in eyes germination, killing of underground sprouts, stem canker and stolon canker resulting in subsequent yield reduction (Banville 1989). The most common effect method for controlling these pathogens is the use of fungicides but the development of resistance in pathogenic fungi to common fungicides and increasing residual hazardous effect on human health and environmental pollution has given a thrust to search for new plant derivatives that can obstruct the fungal Pathogenicity use of natural products for the management of fungal disease in the plants is considered as a good alternate to synthetic fungicides, due to their less negative impact on the environment. Plant extract are not only easy to prepare but also non polluting and low priced as compare to commercial fungicides six plant extract were evaluated for their antifungal activity against Aspergillus niger, Aspergillus flavus, Rhizoctonia solani, Rhizoctonia bataticola. These selected pathogens cause yield losses in numerous economically important crop during vegetative growth. The objective of present study is to see the effect of selected medicinal plant extract against these pathogens. MATERIAL AND METHODS Fair pathogens viz, Aspergillus niger, Aspergillus flavus,

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Rhizoctonia solani, Rhizoctonia bataticola were selected for this experimental work. Pure culture of pathogen was brought from mycology lab and also these were from infected potato, tomato, oil seed like soybean and groundnut with visible symptoms of black scurf of potato. Pathogens were isolated with the help of sterilized forceps and plated on sterilized potato dextrose agar(PDA) medium (potato slice 300 gm, dextrose 20gm, agar 20 gm and distilled water to make the volume 1 liter which was sterilized in autoclave at 15 pounds pressure per sq. inch (PSI) for 20 minutes. Plates were incubated at 25o c and observed daily for emergence of colonies and the pure culture was obtained. Fresh leaves of Azadirachta indica, Aloe vera, Ocimum sanctum, Ocimum basilicum, Lantana camara and Asparagus were collected from local areas. These were washed with tap water and

air dried for one day to eliminate surface moisture. The leaves packed into envelop and kept in oven at 60oc temperature until dried. Dried leaves were grinded separately in mortar and pestle to obtain powder which was then kept in plastic bags for future use. 50 grams of the dried powdered plant were soaked separately in 100 ml water acetone and alcohol respectively extract collected by using soxhlet apparatus. The complete antifungal analysis was carried out under strict aseptic condition. The antifungal activity was evaluated by measuring the radial mycelial growth of fungi.The current study evaluates the effect of dry powder leaf extract of selected plant on growth of mycelium of Aspergillus niger, Aspergillus flavus, Rhizoctonia solani, Rhizoctonia bataticola. OBSERVATIONS

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Table 1. Antifungal activity against Aspergillus niger.

Sl. no. 1 2 3 4 5 6 7 Treatment of dry powder of extract Azadirachta indica Ocimum Sanctum Ocimum basailicum Lantana camara Aloe vera Asperagus Control Colony diameter (mm) of Aspergillus niger Aqueous Acetone Alcohol 25 08 00 35 10 02 25 10 06 35 12 08 25 20 02 35 25 20 45 Inhibition% %Aqueous 100 95.55 86.66 82.22 95.55 55.55

Fig 1. Antifungal activity of different solvent dry powder extract of plant Aspergillus niger. Table 2. Antifungal activity against Aspergillus flavus.
Sl. No Treatment of dry powder of extract Azadirachta indica Ocimum Sanctum Ocimum basailicum Lantana camara Aloe vera Asperagus Control Colony diameter (mm) of Aspergillus flavus Aqueous Acetone Alcohol 30 10 00 35 10 04 30 12 04 40 18 10 30 15 06 40 25 18 50 Inhibition % %Aqueous 100 92 92 80 88 64

1 2 3 4 5 6 7

Fig 2. Antifungal activity of different solvent dry powder extract of plant Aspergillus flavus. Table 3. Antifungal activity against Rhizoctonia solani.
Sl. No Treatment of dry powder of extract Azadirachta indica Ocimum Sanctum Ocimum basailicum Lantana camara Aloe vera Asperagus Control Colony diameter (mm) of Rhizoctonia solani Aqueous Acetone Alcohol 25 08 02 35 10 08 35 12 10 30 12 10 35 06 04 40 25 18 55 Inhibition% %Aqueous 96.36 85.45 81.81 81.81 92.22 67.27

1 2 3 4 5 6 7

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Fig 3. Antifungal activity of different solvent dry powder extract of plant Rhizoctonia solani. Table 4. Antifungal activity against Rhizoctonia bataticola.
Sl. No Treatment of dry powder of extract Azadirachta indica Ocimum Sanctum Ocimum basailicum Lantana camara Aloe vera Asperagus Control Colony diameter (mm) of Rhizoctonia bataticola. Aqueous Acetone Alcohol 35 16 04 30 12 06 32 12 08 35 28 12 40 20 15 45 30 18 55 Inhibition% %Aqueous 92.72 89.09 85.45 78.18 72.72 67.27

1 2 3 4 5 6 7

Fig 4. Antifungal activity of different solvent dry powder extract of plant Rhizoctonia bataticola

DISCUSSION AND CONCLUSION REFERENCES Highest inhibition in radial mycelial growth % induced by Aspergillus niger was exhibited by Azadirachta indica (100 %) followed by Aloe vera (95%) and Ociumum sanctum (95%) (Fig.1) in alcohol (Table 1). While less inhibition in radial mycelial growth was exhibited by Asparagus (Table 1). Maximum inhibition in radial mycelial growth % induced by Aspergillus flavus was exhibited by A.indica (Table 2) followed by O. sanctum and O. basilicum(fig.2). Six plant extract were evaluated for antifungal activity against Rhizoctonia solani A. indica and Aloe vera appeared significantly the most effective and exhibited 90-98 % (Table 3) inhibition in radial mycelial growth of pathogen followed by Ocimum sanctum and Lantana camara (fig.3) Highest inhibition in radial mycelial growth % induced by Rhizoctonia indica (92%) (Table 4) followed by Ocimum sanctum and other plant extract (fig 4). The current results demonstrate that selected medicinal plant extract effective suppressed the radial mycelial growth of all 9 pathogen. In alcoholic extract shows maximum inhibition than acetone and water respectively. [1] Aqsa, Aslam, Farahnaz, Muhammad Arshad, Rahmatullah qureshi and C.A. rauf 2010 In vitro antifungal activity of selected medicanal plant diffusate against Alternaria solai, Rhizoctonia solani and Macrophomina phaseolina, Pak.J.Bot 42(4) 2011-2919. [2] Banville G.J. 1989. Yield losses and damage to potato plants Rhzoctonia solani Kuhn, AM J., 66:821-834. [3] Dubey ,R.C. and Kumar,R Efficacy of Azadirachtin of sclerotia of Macrophomina phaseolina causing charcoal rot in soybean. Indian phytopathology 2003, 56, 216-217. [4] Farr, D., F,Bills chamris and A.Y. Rossman 1989 Fungi on plants and plant products in the United states APS press, st.Paul Minnesota.PP. 1252. [5] Gerretsen, F.C.and N. Haagsma, 1951 accurrence of antifungal substance in Brossica repa, Brassica, Olteracea, and Beta vulgaris Nature (Londan) 168-659.

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[6] Ilyas M.B., Iftikar, K, Anwar, W.and Haq, M. 1997. Effect of different neem products on the vegetative growth and sclerotial production of Macrophomina phaseolina, Pakistan Journal of phytopathology , 9, 77-79. [7] Kannaiyan S.and Prasad, N.N. 1981. Effect of organic amendment on seedling infection of rice caused by Rhizoctonia solani plant and soil , 62, 131-133. [8] Khan, M.W., Kahn A.M. and Saxena S.K. 1974. Rhizospheric fungi and nematodes of egg plant as influenced by oil cake amendments Indian phytopathology , 27, 480-484. [9] Lokhande N.M., Lanjewar, R.D. and Newaskar V.B. 1998. Effect of different fungicides and neem products for control of leaf spot of groundnut J. of soil and crop, 8, 44-46. [10] Madali , N.K. Majumdar, A Chatterjee S.K., Banerjee A. Datla,

J.K. Gupta S. 2009. Antifungal activities and chemical characterization of neem leaf extract on growth of some selected fungal species in vitro culture medium J.Appi s(1), Environ, Manage. Vol 13(1) 49-53. [11] R.C. dubey, Harish Kumar, RR Pandey, 2009. Fungitoxic effect of neem extract on growth and sclerotial survival of Macrophomina phaseolina in vitro,J of American Science 2009 5(5) 17-24. [12] Sharma B.K. and Basandrai, A.K. 1997. Effect of bio control agent, fungicides and plant extract on sclerotial viability of sclerotina sclerotiorum Indian J of Agricultural science, 132133. [13] Singh, R.S. 1986. Incidence of black scurf on potatoes in oil cakes amended soil Indian phytopathology , 35, 300-305.