PHYTOCHEMICAL ANALYSIS AND ANTIFUNGAL POTENTIAL OF DURANTA ERECTA AGAINST SOME PHYTOPATOGENIC FUNGI
HTML Full TextPHYTOCHEMICAL ANALYSIS AND ANTIFUNGAL POTENTIAL OF DURANTA ERECTA AGAINST SOME PHYTOPATOGENIC FUNGI
P. Sharma*, S. Khandelwal, T. Singh and R. Vijayvergia
Plant Pathology and Biochemistry Laboratory, Department of Botany, University of Rajasthan, Jaipur, Rajasthan, India
ABSTRACT
The present study describes the antifungal activity of Durenta erecta L. against some phytopathogenic fungi: Aspergillus niger, A. flavus, A. fumigatus and Penicillium sp. For this purpose, methhanolic extract of leaf, stem and root were prepared and tested by “Disc Diffusion Method”. The methanol extract was subjected to preliminary phytochemical analysis. Alkaloids, saponins, and polyphenols (tannins and flavonoids) were detected as phytoconstituents of the methanol extract. As a result of antifungal activity it was found that the extract of leaf generally revealed antifungal activity against all Aspergillus spp. but activity was highest against A. fumigatus (20±0.67 mm). Stem extract showed less activity against all test fungi but no activity against A. flavus. Root extract did not show any antifungal activity except A. fumigatus with less activity (9±0.98).
Keywords:
Antifungal activity, phytochemical analysis, Duranta erecta L., Phytopathogenic fungi, |
Disc diffusion method
INTRODUCTION: Plants generally produce many secondary metabolites which constitute an important source of microbicides, pesticides and many pharmaceutical drugs. Plant products still remain the principal source of pharmaceutical agents used in traditional medicine 1, 2. Medicinal plants represent a rich source of antimicrobial agents 3. Many of the plant materials used in traditional medicines are readily available in rural areas at relatively cheaper than modern medicines 4.
Duranta erecta Linn. (Syn. Duranta plumieri Jacq., D. repens Linn. and Eng: Goldendewdrop) is commonly known as pigeon berry and locally called ‘Kata mehedi’ belongs to the family Verbenaceae. It is shrubs, herbs or small tree usually 1 to 3 m. in height 5. The plant is not browsed by cattle and is believed to be poisonous 6. Ethyl acetate and aqueous extracts of leaves showed significant antimalarial activity when administered to mice 7.
The fruits are used in the treatment of malaria and intestinal worms 8. The leaves are used in the treatment of abscess 9. From the genus Duranta several iridoid glycosides as durantosides I, II, III, IV, and lamiide were isolated 10, 11. Flavonoids and C-alkylated flavonoids 12, 13 and some alkaloids 14 were isolated.
The aim of the present study was phytochemical screening of plant and evaluation of the in vitro antifungal activity of the crude methanolic extracts (Leaf, stem and root), of Duranta erecta against some phytopathogenic fungi.
MATERIAL AND METHOD:
Plant Material: Duranta erecta leaf, stem and root were collected from local area of Jaipur city. The collected material was authenticated from Herbarium, Department of Botany, University of Rajasthan, Jaipur.
Extract Preparation: The collected materials were washed thoroughly in water, chopped, air dried for a week at 35-40°C and pulverized in electric grinder. For methanolic extraction, powdered dry plant material (50 gm) was extracted with 100 ml methanol for 24 hrs using Soxhlet apparatus. The extracts were filtered and concentrated under vacuum sounding apparatus for 30 min. and the extracts were stored at 4°C.
Phytochemical screening: Phytochemical screening was performed using standard procedures 15, 16.
- Test for terpenoids (Salkowski’s test): To 0.5 g each of the extract was added 2 ml ofchloroform. Concentrated H2SO4 (3 ml) was carefully added to form a layer. A reddish brown colouration of the interface indicates the presence of terpenoids.
- Test for flavonoids: Three methods were used to test forflavonoids. First, dilute ammonia (5 ml) wasadded to a portion of an aqueous filtrate of the extract. Concentrated sulphuric acid (1 ml) was added. A yellow colouration that disappears on standing indicates the presence of flavonoids. Second, a few drops of 1% aluminium solution were added to a portion of the filtrate. A yellow colouration indicates the presence of flavonoids. Third, a portion of the extract was heated with 10 ml of ethyl acetate over a steam bath for 3 min. The mixture was filtered and 4 ml of the filtrate was shaken with 1 ml of dilute ammonia solution. A yellow colouration indicates the presence of flavonoids.
- Test for saponins: To 0.5 g of extract was added 5 ml of distilledwater in a test tube. The solution was shakenvigorously and observed for a stablepersistent froth. The frothing was mixed with3 drops of olive oil and shaken vigorouslyafter which it was observed for the formationof an emulsion.
- Test for tannins: About 0.5 g of the extract was boiled in 10 ml of water in a test tube and then filtered. A few drops of 0.1% ferric chloride was added and observed for brownish green or a blue-black colouration.
- Test for alkaloids: 0.5 g of extract was diluted to 10 ml with acidalcohol, boiled and filtered. To 5 ml of thefiltrate was added 2 ml of dilute ammonia. 5ml of chloroform was added and shaken gently to extract the alkaloidal base. Thechloroform layer was extracted with 10 ml ofacetic acid. This was divided into twoportions. Mayer’s reagent was added to oneportion and Dragendroffs reagent to theother. The formation of a cream (with Mayer’sreagent) or reddish brown precipitate (withDragendroffs reagent) was regarded aspositive for the presence of alkaloids.
- Test for phenol: To estimate total phenols in each of the test sample, the protocol of Bray and Thorpe 17was followed, wherein a standard curve of caffeic acid (a phenol) was prepared A stock solution (100μg/ml) of caffeic acid was prepared in 80% ethanol, from which 0.1 to 0.9 ml was transferred into test-tubes separately and the volume in each case was raised to 1 ml with 80% ethanol. To each of these tubes, 1 ml of Folin-Ciocalteu reagent (prepared by diluting the reagent with distilled water in 1:2 ratio just before use) accompanied by 2 ml of 20% Na2CO3 solution was added and the mixture was shaken vigorously. Each of these were boiled on a water bath (1 min), cooled and diluted to 25 ml with distilled water. The OD was taken at 750 nm using a spectrophotometer against a blank.
- Test for steroids: To 0.2 g of each portion, 2 ml of acetic acid was added; the solution was cooled well in ice followed by the addition of conc. H2SO4 carefully. Colour development from violet to blue or bluish-green indicated the presence of a steroidal ring i.e. aglycone portion of cardiac glycoside 16.
Antifungal activity:
- Test Organisms: Four test organisms, Aspergillus niger A. flavus, A. fumigatus and Penicillium sp. were obtained from Plant Pathology Laboratory, University of Rajasthan, Jaipur, Rajasthan and maintained on Potato Dextrose Agar (PDA).
- Bioassay: Disc diffusion bioassay was employed for testing antifungal activity of plant extracts 18. The readymade PDA medium (Hi-media, 39g) was suspended in 1 lit. distilled water and autoclaved at pressure of 15lbs for 20min. Seven days old cultures of test organisms (0.5 ml) were seeded onto plate and uniformly spread with spreader. Paper discs measuring 6mm diameter, that absorbs about 0.1ml of the test sample and a known quantity of standard reference antibiotic (Fluconazole) were used. The inoculated plates were kept at 5°C for 45-55min and then incubated at 25-27°C for 48hrs. The inhibition zone was measured and compared with those of the standard reference antibiotics. Three to four replicates were maintained for each treatment.
- Determination of activity index: The activity index 19 of the crude plant extract was calculated as:
Activity index (A.I.) = Mean of zone of inhibition of the extract/Zone of inhibition obtained for standard antibiotic drug.
RESULTS AND DISCUSSION: The presence of alkaloids is interesting, as significant quantities are used as antimalarials, analgesics and stimulants 20. The flavonoids are known to inhibit tumor growth and serve also to protect against gastrointestinal infections and are of Pharmacognostic importance thus giving evidence to the use of the plant in ethnomedicine.
Tannins are known to have antimicrobial and astringent activity and play a very important biochemical role in wound healing 21. Such roles include binding to proteins of exposed tissues, thus precipitating the proteins, and forming antiseptic protective coat which enables the regeneration of new tissues to take place. Thus, the antibacterial activity of the extract will have a positive effect on wound healing. In the present work phytochemical screening of the crude extracts of leaf of Duranta erecta revealed the presence of excellent amount of alkaloids, flavonoids, terpenoids, saponins and phenolic compounds (Table 1).
TABLE 1: RESULT OF THE PHYTOCHEMICAL SCREENING OF CRUDE METHANOLIC EXTRACTS OF DURANTA ERECTA.
Phytochemical groups | Leaf | Stem | Root |
Alkaloids | ++++ | + | + |
Flavonoids | +++ | ++ | ++ |
Terpenoids | ++++ | + | + |
Saponins | ++++ | - | - |
Steroids | + | ++ | + |
Tannins | + | + | + |
Phenolics | ++++ | ++ | ++ |
- Absent; + Present; ++ Low concentration; +++ Moderate concentration; ++++ High concentration.
The antimicrobial compounds may be found as alkaloids, flavonoids, phenolic compounds, saponins, steroids, tannins and triterpenoids, whose presence may be attributed to the medicinal properties of plants 22, 23, 24, 25.
In the present study, leaf extract was found to possess higher antifungal activity against all Aspergillus spp. but activity was highest against A. fumigatus (20±0.67 mm). Stem extract showed less activity against all test fungi except A. flavus (No activity). Root extract did not show any antifungal activity except A. fumigatus with less activity (9±0.98) (figure 1, Table 2).
PLATE: A PLATE: B
a: Leaf; b: Stem; c: Root; d: Standard
FIG. 1: ANTIFUNGAL ACTIVITY OF DURANTA ERECTA AGAINST A. FLAVUS (PLATE: A) AND A. FUMIGATUS (PLATE: B)
TABLE 2: ANTIFUNGAL ACTIVITY OF DURANTA ERECTA CRUDE METHANOLIC EXTRACT ON DIFFERENT PHYTOPATHOGENIC FUNGI
Plant parts | A. niger | A. flavus | A. fumigatus | Penicillum sp. | ||||
DIZ (Mean ± SEM mm) | AI | DIZ (Mean ± SEM mm) | AI | DIZ (Mean ± SEM mm) | AI | DIZ (Mean ± SEM mm) | AI | |
Leaf | 17±0.4 | 0.85 | 15±1.32 | 0.42 | 20±1.77 | 1.00 | 7±0.28 | 0.25 |
Stem | 15±1.2 | 0.75 | - | - | 15±0.67 | 0.75 | 10±1.31 | 0.35 |
Root | - | - | - | - | 9±0.98 | 0.45 | - | - |
Fluconazole
(Standard) |
20±0.44 | 17±1.64 | 20±0.54 | 28±1.54 |
DIZ = diameter of inhibition zone; AI= activity index
REFERENCES:
- Ibrahim MB: Anti-microbial effects of extract leaf, stem and root bark of Anogeissus leiocarpus on Staphylococcus aureaus, Streptococcus pyogenes, Escherichia coli and Proteus vulgaris. J. Pharma. Devpt . 1997; 2: 20-30.
- Ogundipe O, Akinbiyi O and Moody JO: Antibacterial activities of essential ornamental plants. Nigeria J. Nat. Prod. & Med. 1998; 2: 46-47.
- Mahesh B, Satish S: Antimicrobial activity of some important medicinal plant against plant and human pathogens. World J. Agri.Sci . 2008; 4 (5): 839-843.
- Mann A, Banso A and Clifford LC: An Antifungal property of crude plant extracts from Anogeissus leiocarpus and Terminalia avicennioides. Tanzania J. Health Res. 2008; 10: 34-38.
- David P: Bengal Plants. Dehradun, India. 1981; 822-838.
- Nelson G: The shrubs and woody vines of Florida. Pineapple Press, Inc. Sarasota 1996.
- Castro O, Barrios M, Chinchilla M and Guerrero O: Chemical and biological evaluation of the effect of plant extracts against Plasmodium berghei. Rev De. Biol. Trop . 1996; 44: 361-367.
- Whistler WA: Tropical ornamentals, a guide. Timber Press, Inc., Portland.2000.
- Xiao P: A pictorial Encyclopaedia of Chinese Medical Herbs, Chuokoron-Sha, Inc.1992.
- Takeda Y , Morimoto Y, Matsumoto T, Ogini C, Hirata E, Takushi A and Otsuka H: Iridiod glycosides from the leaves and stem of Duranta erecta. Phytochemistry. 1995; 39(4): 829-833.
- Rimpler H and Timm H: Iridiods and ecdysones from Verbenaceae. V. Iridiods from Duranta repens L. Z Naturfosch (C).1974; 29:111-115.
- Salama OM, Amer MM, Lahloub MF and Spengel S: Repennoside, A new iridiod glucoside from Duranta repens Fruits. Mans. J. Pharm. Sci . 1992; 8: 212-221.
- Anis, I, Ahmed S, Malik A, Yasin A and Choudary MI: Enzyme inhibitory constituents from Duranta repen. Chem Pharm. Bull. 2002; 50: 515-518.
- Subramanian SS and Nair AGR: Scutellarein and pectolinarigenin from the leaves of Clerodenron phlomides and Duranta repens. Phytochemistry. 1972; 50: 3095-3096.
- Trease GE and Evans WC: Pharmacognosy: A physician guide to herbal medicine, 13th edn. Bailliere Tindall, London. 1989; 176-180.
- Sofowora A: Medicinal plants and Traditional Medicine in Africa. Spectrum Books, Ibadan. 1993; 150.
- Bray HG and Thorpe WV: Analysis of phenolic compounds of interest in metabolism. Meth Biochem. Anal. 1954; 1: 27-52.
- Linday EM: Practical Introduction to Microbiology. (E & FN Spon Ltd). 1962; 77.
- Singh B, Sahu PM and Sharma MK: Anti-inflammatory and Antimicrobial activities of triterpenoids from Strobilanthes callosus Nees. Phytomedicine. 2002; 9:355-359.
- Duke JA and Ayensu ES: Medicinal Plants of China Reference Publications. Inc. 1985; ISBN 0-917256-20-4
- Karodi R, Jadhav M and Bafina A: Evaluation of the wound healing activity of a crude extract of Rubia cordifolia L, (Indian madder) in mice. Int. J. App. Res. Nat. Prod. 2009; 2(2): 12-18.
- Shah, GL, Menon AR and Gopal GV: An account of the ethnobotany of Saurashtra in Gujarat State. J. Econ. Tax. Bot. 1981; 2: 173-182.
- Rabe, T. and Van Stadin VJ: Antibacterial activity of South African plants used for medicinal purposes. J. Ethnopharmacol. 1997; 56: 81-87.
- Ramasamy S: Antibactrial effect of extracts of medicinal plants Anisomeles indica, Anisomeles malabrica, Blumea lacera and Melia azadirachta on selected pathogenic bacteria. M.Phil. Thesis. Submitted to Bharathidasan University, Trichy.2000.
- Santhi R, Alagesaboopathi C, Rajasekara Pandian M: Antibacterial activity of Andrographis ineata Nees and Andrographis echioides Nees of Shevaroy hills of Salem District, Tamil Nadu. Ad. Plant Sci. 2006; 19(II): 371-375.
How to cite this article:
Sharma P, Khandelwal S, Singh T and Vijayvergia R. Phytochemical Analysis and Antifungal Potential of Duranta Erecta against some Phytopatogenic Fungi. Int J Pharm Sci Res 2012; Vol. 3(7): 2686-2689.
Article Information
46
2686-2689
601KB
1440
English
IJPSR
P. Sharma*, S. Khandelwal, T. Singh and R. Vijayvergia
Plant Pathology and Biochemistry Laboratory, Department of Botany, University of Rajasthan, Jaipur, Rajasthan, India
prl.shrm1@gmail.com
24 April, 2012
14 May, 2012
27 July, 2012
http://dx.doi.org/10.13040/IJPSR.0975-8232.3(8).2686-89
01 August, 2012