STUDIES ON ANTIFUNGAL POTENTIAL OF BRYUM CELLULARE (A MOSS) CRUDE EXTRACTS AGAINST SPORE GERMINATION OF FUNGUS CURVULARIA LUNATAHTML Full Text
STUDIES ON ANTIFUNGAL POTENTIAL OF BRYUM CELLULARE (A MOSS) CRUDE EXTRACTS AGAINST SPORE GERMINATION OF FUNGUS CURVULARIA LUNATA
- S. Deora *1 and N. Guhil 2
Department of Botany, New Campus, Jai Narain Vyas University, Jodhpur - 342005, Rajasthan, India.
Department of Botany, B.N.P.G College, Udaipur - 313001, Rajasthan, India
ABSTRACT: Chemical protection plays a decisive role in the resistance of plants against various pathogens and their spore germination. The so-called secondary metabolites which are a characteristic feature of plants and are important to protect plants against a wide variety of microorganisms including fungi. In order to this, the present study was carried out to evaluate the antifungal potential of Bryum cellulare aqueous and ethanolic crude extracts against percentage inhibition of spore germination and hyphal length of test fungi Curvularia lunata, the causal organism of leaf spot of Zea mays. The preliminary tests showed that flavanoid, sterols and terpenoids etc are present in the test plant. Further various concentrations of crude extracts of aqueous and ethanolic form were tested against spore germination and hyphal length of C.lunata through hanging drop method. The results showed that all the extracts posses a significant antifungal potential but highest inhibition of percentage in spore germination was observed in ethanolic extract of B.cellulare. The percentage of spore germination was 60.43 and 15.20 in 10 and 100 per cent concentration of the extract respectively in comparison to control i.e. 110.46 per cent. In aqueous it was 85.13 and 33.26 in 10 and 100 per cent concentration in comparison to its respective control.
Crude extracts, Bryum cellulare, Curvularia lunata, Zea mays
INTRODUCTION: Bryophytes are considered as a ‘remarkable reservoir’ of novel natural products or secondary metabolites, which have shown interesting biological activity and could be used in medicine. Bryophytes especially mosses and liverworts are the source of man biologically active novel compounds pertaining to pharmaceutical uses 1-4. The occurrence of antibiotic substances in bryophytes have been well documented by botanists and microbiologists 5. They possess compounds such as alkaloids (clavatoxine, clavatine, nicotine, lycopodine) polyphenolic acids (dihydrocaffeic) and flavonoids (apigenin, triterpenes etc.) but only few of the species have yet been thoroughly studied.
Recently, public demand of plant base medicine and rise of antibiotic resistant bacteria have motivated biologists to look for new plant based natural products. Therefore, bryophytes can be a promising source of many new biologically active compounds in nature. Apart from the medicinal properties, bryophytes are important component of ecosystem diversity and adds up to the species richness 6, 7, 8 and also increases the plant biomass in forests in some cases 9. Further, bryophytes are important environmental indicators and have been used as predictors of past climate change to validate climate models and potential indicators of global warming 10. They play a chief role in ecosystem functions, such as soil development, nutrient biogeochemical cycling, water retention, plant colonization seed germination, seedling growth, and forest renovation 11.
Deora et al.12 carried out the studies on antibacterial effect of aqueous crude extract of Plagiochasma articulatun, Anthoceros longii, Fissidens bryoides; liverwort, hornwort and moss respectively for their antibiotic effect on Agrobacterium tumifacians and reported that mosses are highly antibiotic in nature followed by hornwort and liverworts. The aim of the present study was to find out the antifungal acitivity of Bryum cellulare against spore germination of phytopathogenic fungi Curvularia lunata. Phytochemical screening of different extract will also carried out to detect the presence of secondary metabolites.
MATERIAL AND METHODS:
Collection of Plant material and extract preparation:
The moss Bryum cellulare was collected in rainy season (2013) from Mt. Abu, Distt. Sirohi (Raj.) around Nakki Lake, Guru Shikhar and Sunset point in both vegetative and sporophytic phases. Both plants were washed with distilled water to remove soil particles, attached litter, dead material. For ethanolic extract preparation, plant material weighted was grinded in mortar and pestle with equal amount of ethanol till the formation of fine paste, then it was centrifuged and filtered. This filtrate was used as (100%) crude extract then it was serially diluted by double distilled water to prepare various concentrations from 10-100 per cent. The same method was adopted for aqueous extract preparation except grinding the plant material with water instead of ethanol.
The test fungi Curvularia lunata (MTCC No.283) was obtained from the Institute of Microbial Technology, Chandigarh, India and sub-cultured in laboratory at 30ºC temperature to obtain its pure isolate.
Screening of antifungal activity:
Fungal spores of the test fungi were bioassayed against the extracts on cavity slides by hanging drop method13. Hyphal length was measured after 8 hrs. of inoculation using Ocular-micrometer14 under Compound Microscope. Percentage of spore germination was counted under light microscope after 24 hrs of incubation.
Phytochemical Analysis: Phytochemical analysis of moss Bryum cellulare extract was done by the methods of Trease and Evans 15 to detect the presence or absence of certain bioactive compounds.
TABLE 1: PHYTOCHEMICAL SCREENING OF THE BRYUM CELLULARE EXTRACTS
|Compound||Ethanolic extract||Aqueous extract|
(+) = phytoconstituents present
(-)= phytoconstituents absent
RESULTS AND DISCUSSION:
Antimicrobial activity of selected bryophyte extracts in different solvents on test microorganism Curvularia lunata are represented in Table 2, 3. The percentage of spore germination and hyphal length of Curvularia lunata was examined using different concentrations of ethanolic and aqueous extracts of Bryum cellulare. The observations on the spore germination indicated the adverse effect of ethanolic extract. The spore germination was decreased from lower to higher concentrations, only 15.20 per cent spore germination reported at 100 per cent concentration while it was 60.43 per cent at 10 per cent. Hyphal length also decreased from 60.40 μm to 0.00 at 10 to 100 per cent concentration (Table 2, Photoplate 2). Percentage of spore germination was 85.13 and 33.26 at 10 to 100 per cent concentration and 121.30 μm hyphal length were observed at 10 per cent while 38.13 μm were noticed in 100 per cent concentration of aqueous extract (Table 3, Photoplate 3). Various extract were tested for the presence of flavanoids, terpenoids, sterols, alkaloids etc. however terpenoids, sterols, flavanoids were detected in extracts of Bryum cellulare (Table 1). The bryophyte extracts prepared in different solvents were found effective in reducing fungal growth as they possess various secondary metabolities which acts as antifungal agent.
The activity of different solvent extracts was in order of ethanolic> aqueous as the bioactive compounds are more soluble in organic solvents. The present results showed similarity with the results of Deora et al.,16 who determined the antifungal activity of a moss against certain phytopathogenic fungi. Deora and Suhalka 17 studied the effect of liverwort R.gangetica against F.moniliforme and found cold water extract was more effective than boiled water extract. Bodade et al, 18 evaluate the antimicrobial efffect of Plagiochasma appendiculatum, Thuidium cymbifolium, Bryum cellulare, Bryum argentium and Racomitrium crispulum on 12 microorganism.
TABLE 2: EFFECT OF ETHANOLIC CRUDE EXTRACT OF BRYUM CELLULARE ON CURVULARIA LUNATA
|SN||Extract Concentration (%)||Spore Germination (%)||Hyphal Length (µm)|
FIG.1: BRYUM CELLULAR ETHANOLIC EXTRACT
TABLE 3: EFFECT OF AQUEOUS CRUDE EXTRACT OF BRYUM CELLULARE ON CURVULARIA LUNATA
|SN||Extract Concentration (%)||Spore Germination (%)||Hyphal Length (µm)|
FIG.2: BRYUM CELLULAR AQUEOUS EXTRACT
CONCLUSION: From the present study it was concluded that spore germination and hyphal length at higher concentration suggests that some antifungal potent chemicals are present in Bryum cellulare which have inhibited the growth. Further this study will also unlock the naturally occurring antifungal phytochemicals in this moss which can be further used as biocontrol agents against plant diseases.
ACKNOWLEGMENT: The authors are thankful to Principal and Head- Department of Botany, Faculty of Science, B.N.P.G College Udaipur (Raj), for providing necessary facilities.
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How to cite this article:
Deora GS and Guhil N: Studies on Antifungal Potential of Bryum Cellulare (A Moss) Crude Extracts against Spore Germination of Fungus Curvularia Lunata . Int J Pharm Sci Res 2016; 7(1): 353-57.doi: 10.13040/IJPSR.0975-8232.7(1).353-57.
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G. S. Deora * and N. Guhil
Department of Botany, New Campus, Jai Narain Vyas University, Jodhpur, Rajasthan, India
20 July, 2015
09 September, 2015
06 November, 2015
01 January, 2016