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ORIGINAL ARTICLE Table of Contents   
Year : 2009  |  Volume : 3  |  Issue : 1  |  Page : 63-65
In vitro antifungal potency of some plant extracts against Fusarium oxysporum


Laboratory of Plant Tissue Culture and Secondary Metabolites (Laboratory No.3), Department of Botany, University of Rajasthan, Jaipur - 302 004, Rajasthan, India

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Date of Submission10-Jul-2008
Date of Acceptance19-Sep-2008
 

   Abstract 

In an approach towards the development of ecofriendly antifungal compounds for controlling plant diseases caused by Fusarium oxysporum , different extracts of three weed plants, namely, Capparis decidua, Lantana camara and Tridax procumbens, were tested for their antifungal potential. The spore germination/spore counting technique was followed for the evaluation of the antibiotic properties of the extracts at three different concentrations. Results revealed that the free flavonoids and sterols of T. procumbens (flower) and bound flavonoids of C. decidua (fruit and stem) totally inhibited spore germination of the fungi (100%). The antifungal components from these plants could be used in developing novel fungicides (biopesticides) for the diseases caused by F. oxysporum (plant pathogen).

Keywords: Capparis decidua, Fusarium oxysporum , Lantana camara , percentage spore inhibition, Tridax procumbens

How to cite this article:
Sharma B, Kumar P. In vitro antifungal potency of some plant extracts against Fusarium oxysporum . Int J Green Pharm 2009;3:63-5

How to cite this URL:
Sharma B, Kumar P. In vitro antifungal potency of some plant extracts against Fusarium oxysporum . Int J Green Pharm [serial online] 2009 [cited 2014 Dec 20];3:63-5. Available from: http://www.greenpharmacy.info/text.asp?2009/3/1/63/49377



   Introduction Top


Fusarium oxysporum also referred to as panama disease or agent green, is a plant pathogenic fungus that causes ' Fusarium wilt' in more than a hundred species of plants such as tomato, potato, sugarcane, cowpea, Musa spp ., pea, ginger, etc. It colonizes the xylem of the host plant, and as a result, blockage and breakdown of the xylem leads to wilt disease symptoms such as, leaf wilting, yellowing and eventually the death of the plant. [1] Management of F. oxysporum is required, as this pathogen and its many special forms affect a wide variety of hosts of economic value.

The development of resistance to common fungicides and increasing restrictions on the use of toxic material in the environment has given an impetus to the search for novel plant protectants that interfere with the fungal pathogenicity factors. Use of natural products for the control of fungal diseases in plants is considered as an alternative to synthetic fungicides, due to their lower negative impacts on the environment. Besides being harmless and nonphytotoxic, it has been proved that plant extracts exhibit effects on germination and on the viability of fungal spores as well. Several higher plants and their constituents have been successful in plant disease control and have proved to be harmless and nonphytotoxic, unlike chemical fungicides. [2],[3],[4] Three weed plants, namely, Lantana camara L. (Verbenaceae), Tridax procumbens L. (Asteraceae) and Capparis decidua Forsk (Edgew) (Capparaceae) were selected in the present study, for evaluation of their antifungal activities. The selected plants were well adapted to harsh (xerophytic) climatic conditions and were well known, among local natives, for their medicinal properties.


   Materials and Methods Top


Different parts of L. camara (root, stem, leaf and flowers), T. procumbens (root, stem, leaf and flowers) and C. decidua (root, stem and fruits) were collected from different localities of Jaipur and its nearby areas. The selected plant parts were separately shade dried, finely powdered and subjected to flavonoid and alkaloid extraction, following the method of Subramanian and Nagarajan [5] and Harborne, [6] respectively. The flowers of T. procumbens were taken for sterol extraction, [7] while the leaves of L. camara were Soxhlet extracted in 80% ethanol for 24 hours, and the ethanolic extract thus obtained was further used for bioassay.

The plant pathogenic microorganism F. oxysporum (MTCC 7678) was procured from the Microbial Type Culture Collection (MTCC), in Chandigarh. The fungal culture was grown and maintained on a Potato Dextrose Agar medium. The antifungal activity of plant extracts (2.5mg/ml, 5mg/ml and 10mg/ml) was investigated by using the method of spore counting. [8] Spore suspension was prepared from a 15-day-old culture of the fungus in sterile distilled water, and 100µl fungal suspension was added to 100µl plant extracts, of concentration 2.5mg/ml, 5.0mg/ml and 10mg/ml (prepared in acetone), in glass vials and incubated at 25 ± 2°C for 24 hours. The control vials contained acetone in place of plant extracts. After incubation, the content of the vials was stained with cotton blue and mounted in lactophenol. The spores were observed under a microscope for their germination status. Percentage inhibition was calculated by using the established formula.



A: Spore inhibition in control; B: Spore inhibition in treatment


   Results Top


The effect of different extracts of selected plants was observed on spore germination of F. oxysporum [Figure 1],[Figure 2],[Figure 3]. A total of 33 extracts were screened for their antifungal properties at three different concentrations (2.5mg/ml, 5.0mg/ml and 10mg/ml). Except for two, all the other plant extracts were found to be significantly active against the selected pathogen, at the three tested concentrations. Twelve extracts showed >80% inhibition in spore germination at 10mg/ml concentration, while 12 extracts inhibited spore germination >70% at 5mg/ml concentration, thus showing significant antifungal potential against the test pathogen. Free flavonoids and sterols of T. procumbens (flower) and bound flavonoids of C. decidua (fruit and stem) totally inhibited spore germination of the fungi (100%) at the concentration of 10mg/ml, while >95% inhibition and >85% inhibition was reported at 5mg/ml and 2.5mg/ml concentrations, respectively. Free flavonoids of L. camara (flower), C. decidua (fruit), T. procumbens (stem, leaf), bound flavonoids of T. procumbens (leaf, flower) and alkaloids of T. procumbens (flower) were found with excellent inhibitory potential against the test pathogen. Observations revealed that the free flavonoids of selected plants were more effective than the bound flavonoids and alkaloids of the plants.


   Discussion Top


In the present investigation the results are quite encouraging, as all the extracts, except two, are found to be active against test plant pathogens and most of the extracts totally inhibited spore germination. For pathogenic fungi, spore germination is a determining factor at the onset of host colonization, and many of the tested extracts have a definite potential, which can be used as a new effective fungicide.

Extracts of T. procumbens were found to have enormous antifungal potential and the plant was observed to be the most potent in the present study followed by C. decidua and L. camara. The selected plants were well known for their antimicrobial properties, but most of the study was carried out with human pathogens. [9],[10],[11],[12],[13],[14]

Application of chemical fungicides was a conventional method to control diseases caused by fungal pathogens, but tremendous health hazards have been reported from time to time during the application of chemical fungicides, in the field conditions. [15] The present investigation has been carried out to explore the hidden antifungal potential of weed plants. Many of the tested plant extracts showed an excellent antifungal efficacy, which may be used for formulating new, safer and ecofriendly fungicides.


   Acknowledgment Top


The authors are thankful to the Head of the Botany Department, University of Rajasthan, Jaipur, India, for providing all the facilities to carry out the study.

 
   References Top

1.Agrios GN. Plant Pathology. 3 rd ed. New York: Academic Press, Inc; 1988. p. 803.  Back to cited text no. 1    
2.Alam S, Akhter N, Begum F, Banu MS, Islam MR, Chowdhary AN, et al . Antifungal activities ( in vitro ) of some plant extracts and smoke on four fungal pathogens of different hosts. Pak J Biol Sci 2002;5:307-9.  Back to cited text no. 2    
3.Dubey RC. Fungicidal effect of essential oils of three higher plants on sclerotia of Macrophomina phaseolina. Indian Phytopathol 1991;44:241-3.  Back to cited text no. 3    
4.Singh RN, Sindhu IR, Gupta K. Effect of leaf exudates and extract of apinach on some phylloplane fungi. Acta Bot Indica 1986;14:104-10.  Back to cited text no. 4    
5.Subramanian SS, Nagarjan S. Flavonoids of the seeds of Crotolaria retusa and Crotolaria striata. Curr Sci 1969;38:65.  Back to cited text no. 5    
6.Harborne JC. Phytochemical Methods: A guide to modern techniques of plant analysis. 2 nd ed. London, New York: Chapman and Hall Ltd.; 1984.  Back to cited text no. 6    
7.Tomita Y, Uomori A, Minato H. Steroidal sapogenin and sterols in tissue cultures of Dioscorea tokora. Phytochemistry 1970;9:111-4.  Back to cited text no. 7    
8.Milosevick T, Slavica S, Sudolak S. In vitro study of ethanolic extract of Hypericum perforatum L. on growth and sporulation of some bacteria and fungi. Turk J Biol 2007;31:237-41.  Back to cited text no. 8    
9.Almas K. The antimicrobial effects of seven different types of Asian chewing sticks. Odontostomatol Trop 2001;24:17-20.  Back to cited text no. 9    
10.Ali-Shtayeh MS, Abu Ghdeib SI. Antifungal activity of plant extracts against dermatophytes. Mycoses 1999;42:665-72.  Back to cited text no. 10    
11.Sukul S, Chaudhuri S. Antibacterial natural products from leaves of Lantana camara L. Indian J Pharm Sci 2001;63:20-3.  Back to cited text no. 11    
12.Alves TM, Silva AF, Brandγo M, Grandi TS, Smβnia E, Smβnia Júnior A, et al . Biological screening of Brazilian medicinal plants. Mem Inst Oswald Cruz 2000; 95:367-73.  Back to cited text no. 12    
13.Taddie A, Rosas-Romaro AJ. Bioactivity studies of extracts from Tridax procumbens . Phytomedicine 2000;7:235-8.  Back to cited text no. 13    
14.Udupa SL, Udupa AL, Kulkarni DR. Influence of Tridax procumbens on lysyl oxydase activity and wound healing. Planta Med 1991;57:325-7.  Back to cited text no. 14    
15.Alam S, Alam MS, Mahal F. Growth inhibition ( in vitro ) of chilli fruit rot pathogen Alternaria tenuis. J Asiat Soc Bangla Sci 1999;25:211-6.  Back to cited text no. 15    

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Correspondence Address:
Bindu Sharma
Laboratory of Plant Tissue Culture and Secondary Metabolites (Laboratory No.3), Department of Botany, University of Rajasthan, Jaipur - 302 004, Rajasthan
India
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DOI: 10.4103/0973-8258.49377

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    Figures

  [Figure 1], [Figure 2], [Figure 3]

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    Abstract
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    Materials and Me...
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