ABSTRACTThe investigation was aimed to carry out the antifungal activities of the methanol and aqueous extracts of Abutilon theophrasti against four fungalstrains (Microsporum gypseum, Penicillium spp., Fusarium spp., Aspergillus spp.). Antifungal activity of the extracts was determined by the discdiffusion method against various fungi with soubred dextrose agar as media. Methanolic extracts of A. theophrasti showed promising antifungalactivity compared to aqueous extracts against selected fungal species. Methanolic extract of leaf displayed the highest activity with zone of inhibition14 mm against Penicillium spp. which were totally concentration dependent; however, a poor activity was seen in aqueous extracts. Methanolicextracts of root displayed good antifungal activity, while as no activity was seen in aqueous extracts of root. The results obtained in the study showthat methanolic leaf extracts possess phytoconstituents with potent antifungal activity.Keywords: Abutilon, Methanol, Rhizopus, Penicillium

Habib, Huma

Hassan, Musheerul

Kausar, Nasreen

Ma, Zargar

Mir, Reyaz Ahmad

English

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Vol 9, Issue 3, 2016Online - 2455-3891 Print - 0974-2441EVALUATION OF ROOT AND LEAF OF ABUTILON THEOPHRASTI MEDIK FOR ANTIFUNGAL ACTIVITYMUSHEERUL HASSAN1, HUMA HABIB2, NASREEN KAUSAR3, ZARGAR MA4, REYAZ AHMAD MIR5*1Department of Biotechnology, Pacific University, Udaipur, Rajasthan, India. 2Department of Biochemistry, Islamia College of Science and Commerce, Srinagar, Jammu and Kashmir, India. 3Department of Zoology, University of Kashmir, Srinagar, Jammu and Kashmir, India. 4Department of Biotechnology, Central University of Kashmir, Srinagar, Jammu and Kashmir, India. 5Department of Zoology, Islamia College of Science and Commerce, Srinagar, Jammu and Kashmir, India.  Email: mn-reyazap@gmail.comObjective: To investigate the  antifungal  activity of root and leaf extracts of Abutilon theophrasti against Microsporum gypseum, Penicillium spp, Fusarium spp, Aspergillus spp.Methods: Abutilon theophrasti (velvetleaf) was collected from “Lower Munda” Dist: Qazigund (Jammu & Kashmir) India. The collected samples were dried and extracted with two different polaristic solvents (Methanol and aquous).Antifungal activity of the extract was determined by disc diffusion method against four fungal strains with soubred dextrose agar media.Results: Methanolic extracts of Abutilon theophrasti showed promising antifungal activity compared to aqueous extracts against selected fungal species. Methanolic extract of leaf displayed highest activity with zone of inhibition 14mm (ZOI-14) against Penicillium spp, however poor antifungal activity was observed in aqueous extracts. Methanolic extracts of root displayed good antifungal activity, while as no activity was seen in aqueous extracts of root.Conclusion: It can be concluded from present study that methanolic leaf extracts possess phytoconstituents with potent antifungal activity.Keywords: Abutilon, Methanol, Rhizopus, Penicillium.INTRODUCTIONPlants produce a diverse assortment of organic compounds the majority of which do not participate directly in growth and development of the plant. These substances traditionally referred as secondary metabolites; there functions many of which remains unknown. On other hand metabolites products such as phytosterols, lipids, nucleotides, amino acids, and organic acids are found in all plants and perform an essential and evident role in growth and development. The secondary metabolites are distributed among limited taxonomical groups within the plant kingdom. The complexity of the chemical structures and biosynthetic pathway, natural products have been widely perceived as biologically insignificant and receive less attention from most of the biologists. Organic chemicals are inherited in novel photochemicals and investigating their chemical properties. Abutilon (L) genus of the Malvaceae family comprises about 150 annual or perennial herbs, shrubs or even small plants widely distributed in the tropical and subtropical countries of Asia, Africa, America, and Australia [1].China is stated to be the origin of Abutilon theophrasti. In china, it has been grown since around 2000 BC for its jute like fiber. It is also known by many vernacular names like “velvetleaf, China-jute, button-weed.” A. theophrasti is a quantitative short-day herbaceous annual plant that colonizes highly different habitat such as agriculture fields and high altitude areas of work in Kashmir valley. Time of flowering is mostly rapid under short day photoperiods. Under glass house conditions, increased height, fruit weight, internode length with increasing photoperiods is seen [2]. Nurse et al. (2004) [3] studied the effects of varying natural photoperiods on the germinability of A. theophrasti seeds and seedlings vigor as measured by initial rate of radical growth. A medicinal herb contains a number of chemical compounds called secondary metabolites, whenever any plant found to be useful it is taken up for investigation, as regards to the constituents present for its biological action. On confirmation of its biological activity, the suitable phytochemicals are isolated and put into usage. Many consumers preferred to treat themselves with phytopharmaceuticals or herbal preparation, and the sale of these is increasing in mostly first world countries. All these have led to the development of new field called “Herbal Drugs Extraction.” Many manufacturers are making efforts to improve yields as well as the composition of total extracts and currently also the area of investigating and isolating the herbal drugs is gaining considerable importance and leading to evaluation of green chemicals. Various species of Abutilon are traditionally claimed for their varied pharmacological and medicinal activities. Different parts of Abutilon plant contain specific phytoconstituents responsible for these activities. Kremer (1986) [4] studied antimicrobial activity of A. theophrasti seeds and he found significant antimicrobial activity. Saini et al. (2014) [5] evaluated antifungal activity of Abutilon indicum. Traditionally, Abutilon is used for the treatment of various ailments. The roots of Abutilon are considered useful as Demulcent, Diuretic, in chest infections and urethritis. The decoction of leaves is used in toothache and for inflammation of the bladder. The bark is used as anthelmintic, laxative, and diuretic [6]. The Ayurvedic pharmacopoeia of India indicates the use of roots in Gout, polyuria, piles, gonorrhea and hemorrhagic diseases [7]. Aerial parts of the plant are used in folk medicine as an expectorant and emollient [8]. Besides this A. theophrasti is also claimed to have cytotoxic and antioxidant activities.METHODSCollection of plant materialA. theophrasti (velvetleaf) was collected from “Lower Munda” Dist: Qazigund (Jammu and Kashmir, India). Latitude 33.56° and longitude 75.20°. The plant was identified and registered (VS-No.2113-KASH) at Herbarium center for Biodiversity and Taxonomy, “University of Kashmir” India.Preparation of extractsThe plant material was dried in shade at room temperature for about 15 days. Dried plant samples were powdered by the mechanical grinder. The powder was then sieved to fine mesh, stored in polythene bags at Research ArticleReceived:  8 December 2016,  Revised  and  Accepted:  21 December 2016         ABSTRACTAsian J Pharm Clin Res, Vol 9, Issue 3, 2016, 75-77 Hassan et al. 76room temperature before extraction. Methanol and distilled water were used as solvents. Extraction was done by simple maceration process. The extracts were concentrated to dryness using rotary evaporator and crude extracts were tested for antifungal activity.MicroorganismsFour fungal strains were used, (Microsporum gypseum, Penicillium spp., Fusarium spp., Aspergillus spp.).Determination of antimicrobial activityDisk diffusion method was used to evaluate antifungal activity [9,10]. Standard size Whatman No. 1 filter paper discs, 5.0 mm in diameter, sterilized by moist heat at 121 lb in an autoclave for 15 minutes were used. Saboured dextrose agar (SDA) medium was prepared for disc diffusion test. After sterilization, SDA was poured in sterilized Petri plates and allowed to solidify. A fresh culture of fungi was used for inoculum preparations. Turbidity suspension was taken 0.5 McFarland. Using a sterile cotton swab, fungal cultures were swabbed on the surface of sterile agar plates. The dried plant extracts were re-suspended to 500, 250, and 100 μg/ml in dimethyl sulfoxide and sonicated to dissolve. Sterile 5 mm discs were impregnated with 50 μl of extract and placed on the surface of agar plates inoculated with a microbial culture. Each extract was tested in triplicate. Itraconazole (30 μg/disc) was taken as the reference standard. The plates were incubated at 27°C for 24 hrs. The diameter of the inhibition zones was measured in millimeter. Three replicates were kept in each case and average values were calculated.RESULTS AND DISCUSSIONThe result obtained from antifungal activity assay using methanolic, and aqueous crude extracts showed that growth of all fungi was inhibited (Tables 1 and 2). Methanolic leaf extracts showed good antifungal activity while as very little activity was seen in aqueous crude extracts. Among all selected fungal species methanolic leaf extracts showed the highest activity against Penicillium (ZOI-14) fallowed by M. gypseum (ZOI-11). The present study successfully evaluated the role of A. theophrasti for its antifungal activity. It has been already reported that the phytochemicals have an excellent activity to get against microorganisms [11]. This could be due to active chemicals which are present in A. theophrasti making it a potential antifungal plant. The Table 1: Antifungal activity of methanolic and aqueous extracts of leaf (A. theophrasti)Serial numberFungi Zone of inhibition (mm of diameter)Concentration of methanolic leaf extractsConcentration of aqueousleaf extractsItraconazole100 μg/ml 250 μg/ml 500 μg/ml 100 μg/ml 250 μg/ml 500 μg/ml 30 μg/ml1 Micro sporum gypseum 1 6 11 0 0 2 152 Penicillium spp. 2 8 14 0 0 0 163 Fusarium spp. 0 2 8 0 0 3 174 Aspergillus spp. 2 7 10 0 0 2 16A. theophrasti: Abutilon theophrastiTable 2: Antifungal activity of methanolic and aqueous extracts of root (A. theophrasti)Serial numberFungi Zone of inhibition (mm of diameter)Concentration of methanolic root extractsConcentration of aqueousroot extractsItraconazole100 μg/ml 250 μg/ml 500 μg/ml 100 μg/ml 250 μg/ml 500 μg/ml 30 μg/ml1 Microsporum gypseum 0 0 0 0 0 0 152 Penicillium spp. 0 3 7 0 0 0 163 Fusarium spp. 0 0 5 0 0 0 174 Aspergillus spp. 2 3 8 0 0 0 16A. theophrasti: Abutilon theophrastiFig. 1: 2Asian J Pharm Clin Res, Vol 9, Issue 3, 2016, 75-77 Hassan et al. 77investigated aqueous and methanolic root extracts of A. theophrasti did not show strong antifungal activity, however, negative results do not mean absence of bioactive constituents. Active compounds may be present in insufficient quantities in the crude aqueous extracts to show the activity with the dose level employed. Saini et al. (2014) evaluated the antifungal activity of A. indicum in which he observed the potential antifungal activity in alcoholic extracts compared to aqueous extracts which is in harmony with the present study. Pundair and Sharma [12] have earlier reported that ethanol garlic extracts showed antimycotic activity against fungal genera such as Aspergillus, Rhizopus, Fusarium, and Cladosporium which is in accordance with our present study.CONCLUSIONThis study has demonstrated that A. theophrasti exhibited a potential degree of antimycotic activity. The comparable activity of different extracts can be attributed to polyphenolic compounds.ACKNOWLEDGMENTThe authors are grateful to IIIM Srinagar (Jammu and Kashmir, India), Department of Biotechnology and Allied Sciences, Suresh Gyan Vihar University, Jaipur (Rajasthan) for providing the facilities to complete the work. REFERENCES1. Sikorska M, Matlawska I. Polyphenolic compounds from Abutilon grandiflorum leaves. Acta Pol Pharm 2008;65:467-71.2. Patterson DT. Effect of photoperiods on reproductive development in velvetleaf. Weed Sci 1995;43:627-63.3. Nurse RE, Ditommaso A, Ramirez RA. Planting date effects on the germ inability and seedling vigour of Abutilon theophrasti (Malvaceae) seeds. Phytoprotection 2004;85:161-8.4. Kremer JR. Antimicrobial activity of velvetleaf (Abutilon theophrasti) seed. Weed Sci 1986;34:617-22.5. Saini A, Sin DP, Gutum GK, Ashfaq M, Dwivedi B. Evaluation of root and leaf extract of Abutilon indicum Linn for antifungal activity. Int J Chem Pharm Sci 2014;3(1):717-21.6. Kumar GG, Gali V. Phytochemical screening of Abutilon muticam and Celosia argentea Linn. Int J Pharm Biosci 2011;2(3):463-7.7. Ayurvedic Pharmacopoeia Committee, India. Sahrcara (whole plant). The Ayurvedic Pharmacopoeia of India. Part 1, Vol. 1. New Delhi: Department of ISM and H, Government of India, Ministry of Health and Family Welfare; 1978. p. 25-28.8. Kiyamova SE, Khidyrova NK, Kukina TP, Shakhidoyatov KM. Neutral constituents from leaves of plant of the family Malvacea. I. Abutilon theophrasti. Chem Nat Compd 2012;48(2):297-8.9. Rios JL, Recio MC, Villar A. Screening methods for natural products with antimicrobial activity: A review of the literature. J Ethnopharmacol 1988;23(2-3):127-49.10. Kim DJ, Chung SG, Lee SH, Choi JW. Relation of microbial biomass to counting units for Pseudomonas aeruginosa. Afr J Microbial Res 2012;6(21):4620-2.11. Murala Khrishna T. In vitro determination of anti-oxidant and anti-bacterial of Vitex negunndo Linn. Int J Pharm Bio Sci 2013;4(1):121-7.12. Pundair RK, Sharma JP. Antimicrobial activity of ethanolic extracts of Syzygium aromaticum and Allium satium against food associated bacteria and fungi. Ethnobot Leafl 2010;2010(14):344-60.

EVALUATION OF ROOT AND LEAF OF ABUTILON THEOPHRASTI MEDIK FOR ANTIFUNGAL ACTIVITY

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EVALUATION OF ROOT AND LEAF OF ABUTILON THEOPHRASTI MEDIK FOR ANTIFUNGAL ACTIVITY

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