36th Ann. Meeting of MSI & Nat. Seminar on Fungal Biodiversity & Bioprospecting….. Goa, India Oct., 29-30, 2009
POSTER SESSION 4
PS – 41
Anti-Staphylococcus aureus Activity of Phellinus merrillii
Abdulghani Ameri*, Jitendra G. Vaidya and S.S.Deokule
Department of Botany, University of Pune, Pune 411 007.
*E-mail: ghaniameri@yahoo.com
Development of antimicrobial resistance of numerous bacteria necessitates the need for new and inexpensive drugs that will be able to act for a longer period before resistance sets in. Methicillin resistant Staphylococcus aureus (MRSA) has become a multi-drug resistant to several conventionally prescribed antibiotics. In this study, the antimicrobial potencies of fifteen strains of the mushroom Phellinus were evaluated against 30 strains of clinical isolates of methicillin resistant Staphylococcus aureus using different organic extractives. Phytochemical analysis of the extracts was carried out by thin – layer chromatography.Various polysaccharides fractions (I, II, IIIa and IIIb), diterpenoids, triterpenoids and sesquiterpenoids were isolated and tested against test microorganisms. Chloroform extract of P.merrillii was more effective as compared to other solvents. The minimal inhibitory concentration of the chloroform extract was determined to be 850 µg/ml. Sesquiterpenoids were found to be the most effective component and were subjected to column chromatography.
PS – 42
Antifungal potentiality of essential oil from discarded leaves
Ahsan Kamran*, Piyush Mishra, Rohit Mishra, Amit Tiwari and Anupam Dikshit
Biological Product Laboratory, Department of Botany, University of Allahabad, India 211 002.
*E-mail: drahsan.kamran@ymail.com
Synthetic antimycotic drugs are applied for the control of pest and diseases of the agricultural food commodities and human beings as these are effective, dependable, and economic. However indiscriminate use has resulted in several problems such as pest resistance to pesticides, resurgence of pests, toxic residues in food, causing health hazards to animal and human beings, to water, soil, air, and disruption of ecosystem. Natural products are an alternative to the use of these synthetic pesticides. Medicinal plants are traditional, accessible and affordable source of primary healthcare. Traditional medicines are culturally familiar, financially affordable and simple to use and are assuming a great significance globally as green drugs, which are healthier and safer than synthetic drugs. During the present investigations, in-vitro studies of essential oil obtained from waste guava leaves was done against fungal pathogen of stored grains such as Penicillium expansum and Aspergillus flavus. Their MIC in various media was determined. They were found to be very effective with a lower MIC as compared to various synthetic antifungal.
PS – 43
Mushroom Cultivation and processing for the empowerment of tribal women in Madhya Pradesh
Jaya Singh, Divya Bajpai and A. K. Pandey
Mycological Research Laboratory, Dept. of Biological Sciences,
Rani Durgawati University, Jabalpur (M.P.)
Cultivation of edible mushroom is an ideal income generating activity for the tribal/rural women and unemployed youth. Labour intensive nature, significant value addition, dependence on locally available, abundant and cheap raw material (straw), low initial investment, simple technology and limited space requirement make it a worth while proposition. Women can be effective mushroom growers as long as they are provided with support services and appropriate training from mushroom development centers. With the right training women can go in to this venture and supplement their family income who contributes a high percentage of the labour in the field, especially in the developing countries. Moreover, supplementation of traditional diet of rural population in general and of women and children in particular which is based on millets and coarse grains, with highly proteinaceous mushroom like Pleurotus florida, will help in alleviating the protein crisis in these weaker section of society. The transfer of mushroom biotechnology to the sites of rural women, tribals and aborigines, promises to provide not only a low cost nutrition in the form of fungal protein but will also open avenues for self employment and income generation in tribal areas.
PS – 44
Biosynthesis of Au and Ag nanoparticles using edible mushroom extract
N. Raaman*, R. Jegadeesh, L. Hariprasath, G. Rajasekhar reddy and S. Archana
Fungal Biotechnology, Natural Products and Tissue Culture Laboratory, Centre for Herbal Sciences, Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai 600 025.
*E-mail: raaman55@gmail.com
The biosynthesis principle of nanotechnology is one of the key issues in nanoscience research. There is growing need to develop ecofriendly benign metal nanoparticle synthesis process that do not use toxic chemicals in the synthesis protocols to avoid adverse effects in medical applications. The present work is a report on extracellular synthesis method for the preparation of gold and silver nanoparticles in water, using the extract of Pleurotus djamor var. roseus a naturally occurring edible mushroom, as reducing and capping agents. Gold nanoparticles showing different sizes (20–100 nm) and shapes from triangular nano prisms to nearly spherical and hexagonal were obtained. The silver nanoparticles were spherical of 15-50 nm in size. There was increased productivity of nanoparticles as shown by sharp and intense surface plasmon resonance bands for the nanoparticles prepared using an excess of the extract. All the synthesized nanoparticles were characterized by Ultra violet-Visible spectroscopy and Transmission Electron Microscopy. The protein binding was confirmed by Fourier Transform-Infrared spectroscopy measurements and these results were carried out to identify the possible biomolecules responsible for capping and efficient stabilization of the nanoparticles. It was found that gold nanoparticles were bound to proteins through free amino groups and silver nanoparticles through the carboxylate group of the amino acid residues. The position and intensity of the emission band was found to depend on composition of the nanoparticles indicating the possible use in therapeutic applications. Antibacterial and antioxidant activity were carried out for the nanoparticles.
PS – 45
Bioassay-guided fractions of novel sesquiterpene (3-(4’-hydroxyphenyl-2, 4, 7-trioxa-bicyclo [4, 1, 0] heptane) from Pestalotiopsis microspora Jeon. in Modified PDB culture
S. Kamalraj*, M. Murugan and J. Muthumary
Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai 25.
*E-mail: k_raj822001@yahoo.co. in
Pestalotiopsis sp. was isolated as an endophytic fungus from Mangifera indica L. The Pestalotiopsis microspora is a microbial factory of bioactive secondary metabolites. These strains of Pestalotiopsis microspora differ widely in the degree and diversity of secondary metabolites that they may produce bioactive compounds like Taxol, jestrone, ambuic acid, torreonic acid, pestaloside, pestalotiopsins and 2-γ-hydroxydimeniol (Strobel, et al 2002).The crude extract of the fungus showed significant antimicrobial activity against Micrococcus luteus, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Bacillus subtilis and Candida albicans. Bioassay guided fractionation of this extract was identified as a new sesquiterpene by spectral analysis. An ecological awareness of the role played by this organism in nature will provide the best clues for targeting particular types of endophytic bioactivity with the greatest potential for bioprospecting.
PS – 46
Isolation, Identification and Molecular analysis by 18S rRNA sequence of Taxol producing fungus Pestalotiopsis guepinii (Desm.,)
S. Malathi*, S. Visalakchi and J. Muthumary
Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai 600 025.
*E-mail: maalathi_selvam86@yahoo.co.in, mm_j@rediffmail.com
Taxol producing fungus Pestalotiopsis guepinii (Desm.,) was isolated from the medicinal plant Psidium guajava Linn. The presence of taxol was confirmed by TLC, UV and HPLC. The fungus was identified based on colony characteristics, morphology of conidia and also by 18S rRNA sequencing Phylogenetic tree was obtained and the evolutionary relationship between different organisms were compared. Pestalotiopsis guepinii (Desm.,) was found to be closely related to 18 of the Pestalotiopsis species showing 99% homology.
PS – 47
In-vivo and in-vitro studies on anticancer activity of fungal taxol isolated from an endophytic fungus Botryodiplodia theobromae Pat.
M. Pandi*, P. Rajapriya** and J. Muthumary
Centre for Advanced Studies in Botany, University of Madras, Guindy campus, Chennai 600 025.
**Department of Biotechnology, Srinivasan College of Arts and Science, Perambalur 621 212.
*E-mail: an_pandi@rediffmail.com
Breast cancer is one of the most common cancers in women of developed countries. In India, it is the second most frequent cancer with age standardized rates varying from 22 to 28 per 10,000 women. The drug taxol has been hailed by many in the cancer community as a major breakthrough in the treatment of cancer. It has already been approved in use against ovarian and advanced breast cancer in many countries world- wide. The most common source of Taxol is the bark of trees belonging to the Taxus family including Yew trees. Unfortunately, these trees tend to be rare, slow growing, and a large amount of bark may have to be processed to obtain a small amount of the drug. Alternative sources of Taxol have been sought, and Strobel et al., were the first to isolate Taxol-producing fungi from the tree Taxus brevifolia. Over the last decade there has been a great deal of interest in finding other fungi that produce Taxol. However, very few studies are reported with reference to the cytotoxic effects and anticancer activity of fungal Taxol. Hence, the present study is to investigate the In-vivo and In-vitro effect of fungal Taxol derived from a novel endophytic fungus Botryodiplodia theobromae Pat.
PS – 48
Molecular characterization and antimicrobial properties of hybrid from interspecific hybridization between Pleurotus ostreatus var. florida and P. djamor var. roseus
P. Selvakumar, S. Rajasekar, A. Giridhar Babu**, K. Periasamy, N. Raaman* and M. Sudhakar Reddy**
Fungal Biotechnology, Natural Products and Tissue Culture Laboratory, CAS in Botany,
University of Madras, Guindy campus, Chennai 600 025.
**TIFAC- CORE in Agro & Industrial Biotechnology, Thapar University, Patiala 147 004.
*E-mail: raaman55@gmail.com
Protoplast fusion is an important biotechnological tool to improve the mushroom strains for high yield, quick spawn run, tolerate the adverse conditions, utilize huge variety of agricultural waste, increase in nutritive values and medicinal properties. In the present study, basidiomata of P. ostreatus var. florida (white colour, high yielding mushroom) and P. djamor var. roseus (pink-rose colour, attractive and leathary mushroom) were collected from the natural habitat. To improve the edibility, biological efficiency and medicinal property of P. djamor var. roseus, protoplast fusion was developed between P. ostreatus var. florida and P. djamor var. roseus. The colony morphology, cultural character and fruiting body morphology of hybrid was characterized; it showed intermediate to parental morphology. The RFLP pattern of rDNA (ITS region) of hybrid showed different from the parental Pleurotus spp. In phylogenetic analysis, ITS sequence data of hybrid was closely clad with variety of Pleurotus spp. and designated as Pleurotus sp. HS. The hybrid strain showed high biological efficiency than the parental strains in paddy straw substrate followed by sugarcane bagasse, cotton waste, coir pith and wood chips. The proximate composition, energy values, vitamins and mineral content of hybrid and parental strains were analyzed and hybrid showed rich nutritive value. The antimicrobial properties of hybrid and parental strains were tested by different organic solvent extraction method. Ethyl acetate extract was effective and inhibited the growth of human pathogenic bacteria and yeast. The bioactive compound was isolated and purified by different chromatographic techniques. The minimal inhibitory concentration (MIC) of the purified bioactive compound 5,6,11,12, 13,15,16,17-octahydro-6,11-dihydroxy-4,4,10,13-tetramethyl-17-((E)-5,6-dimethylhept-3-en-2-yl)-4H - cyclopenta[a]phenanthren-3(9H,10 H,14H)-one was tested and it showed good antimicrobial activity.
PS – 49
Fungi: an importunate source of novel antifungal agents
Shilpa Verekar and S. K. Deshmukh
Department of Natural Products, Piramal Life Sciences Limited, 1, Nirlon Complex, Off Western Express Highway, Near NSE Complex, Goregaon (East), Mumbai 400 063.
The search for new drugs against fungal infection is a major challenge to current research in mycotic diseases. The need for new, safe and more effective antifungals is a major concern today, especially with the increase in opportunistic infections in the immunocompromised host. The presentation will cover the current scenario of antifungal antibiotics and the need and approaches to search for newer antifungals antibiotics and drug targets. Fungal diversity of India can be used to get novel diverse secondary metabolites for antifungal. The antifungal compounds isolated from Indian fungi are Mulundocandin and Deoxymulundocandin, Aranorosin, Fumifungin etc. The need of fungal culture collections is highlighted.
PS – 50
Taxol producing a novel endophytic fungus pestalotiopsis funerea from Araucaria bidwilli
K. Srinivasan*, S. Visalakchi and J. Muthumary
CAS in Botany, Guindy Campus , University of Madras, Chennai 600 025.
*E-mail: vksrini267696@yahoo.co.in
Taxus brevifolia, commonly known as the Pacific yew, is a member of the yew family (Taxaceae). It is a small, slow growing evergreen tree native to the northwestern United States (Wheeler et al., 1992). In 1971, a natural product called taxol that was isolated from T. brevifolia. Taxol has been found to be useful in the treatment of various cancers. Endophtic fungi living within the tissues of host plants , and have proven to be rich sources of bioactive natural products as one class of the most widely distributed endophytic fungus pestalotiopsis have attracted much attention in recent years for their ability to produce a variety of new bioactive secondary metabolites.The present study, pestalotiopsis funerea isolated as endophytes from Araucaria bidwilli (Bhutan) was screened for taxol production and fungal diversity, the results will be discussed later
PS – 51
Screening for extracellular enzymes from endophytic fungal strains isolated from medicinal plants
S. Sunitha, D. Nirmala Devi. and C. Srinivas
Department of Microbiology and Biotechnology, Jnana Bharathi Campus,
Bangalore University, Bangalore 560 056.
Endophytic fungi exhibit a complex web of interactions with host plants and therefore they have been intensively studied over the last several years as prolific sources of new bioactive natural products. Fungal enzymes are used in food, beverages, confectionaries, textile and leather industries to simplify the processing of raw materials. Fungal enzymes are gaining importance in industry, agricultural and human health, as they often are more stable than the enzymes derived from other source. The enzyme tests indicate that most endophytes are degraders of the sugars and cellulose available in the host plants. Some of the lginolytic enzymes such as laccase, tyrosinase and peroxidase are helpful in characterization of isolates. The use of simpler solid media permits the rapid screening of large populations of fungi for the presence or absence of specific enzymes. The present study was carried out to find new sources of valuable extracellular enzymes from endophytic fungi. Twenty-five fungal strains, isolated from medicinal plants (Calophyllum inophyllum, Catharanthus roseus, Alpinia calcarata, Carrissa carandus) were screened for extracellular enzymes such as amylase, cellulase, laccase, lipase, pectinase and protease on solid media. 80% of fungi screened showed positive for lipase, 60% for pectinase, 56% showed for amylase, 24% showed for cellulase and Laccase and only 8% showed positive for protease.
PS – 52
Screening of endophytic fungi for the production of Camptothecin an novel anticancer drug
J. Swetha, R. D. Varuni, P. Sindhu and K. P. Kannan*
Bannari Amman Institute of Technology, Sathyamangalam, Erode District .
*E-mail: kp_kannan2001@yahoo.co.in
The nature and biological role of endophytic fungi with their plant host is variable. Endophytic fungi are known to have mutualistic relations to their hosts, often protecting plants against herbivory, insect attack or tissue invading pathogens; and in some instances the endophyte may survive as a latent pathogen, causing or quiescent infections for a long period and symptoms only when physiological or ecological conditions favors virulence. Some of these endophytes produce various useful bioactive molecules and this has encouraged a worldwide scientific effort to isolate and study them. Many reports showed that in microbe-plant relationships, endophytes contribute substances that possess various types of bioactivity such as antimicrobial and antifungal agents. In the present investigation endophytic fungi were isolated from selective medicinal plants and were screened for the production of Camptothecin an anticancer drug. The results will be discussed in detail.
PS – 53
Comparative antimicrobial activity of herbal extracts and cough syrups on upper respiratory tract microbes
K. N. Varalakshmi, C. G. Sangeetha
Department of Biotechnology, Centre for Post-Graduate Studies, Jain University,
#18/3, 9th Main, III Block, Jayanagar, Bangalore 560 011.
The agar well diffusion method was used for screening the antimicrobial activity of four commonly used herbal extracts viz., Oscimum santum (Tulsi), Coleus aromaticus (Doddapatre), Curcuma longa (haldi) and Alium sativum (garlic) to treat cough and were compared to that of commercial cough syrup formulations against four fungi viz., Aspergillus niger, Candida albicans, Fusarium sp. and Penicillium sp. and four bacteria: Enterobacter aerogenes, Staphylococcus aureus, Escherichia coli and Bacillus subtilis. Among the herbal extracts, Tulsi showed maximum inhibition of C. albicans (23.6mm) followed by Doddapatre (17.3mm) and Haldi (10.3mm). Garlic could not inhibit the growth of C. albicans but both Fusarium sp. and Penicillium sp. were inhibited (8.6mm). Among the cough syrups, Ambrodylite was most effective in inhibiting C. albicans followed by Himalaya koflet and Benadryl. Salmucolite was least effective in inhibiting the fungi. Antibacterial activity was more for the commercial cough syrups than that for the herbal extracts. Benadryl was most effective followed by Himalaya koflet and Salmucolite. Among the herbal extracts, only Garlic was effective in inhibiting the growth of E. aerogenes and E. coli whereas, Tulsi and Doddapatre could inhibit the growth of only E. coli. It was found that herbal extracts were more effective in inhibiting the fungal pathogens than the commercial cough syrup formulations. From this study it can be concluded that local home remedies could be more beneficial in treating cough.
PS – 54
A novel endophytic anticancer compound producing fungus Alternaria sp. SVJM05 isolated from a medicinal plant, Indigofera enneaphylla Linn.
S. Visalakchi*, K. Srinivasan and J. Muthumary
Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai 600 025.
*E-mail: svisalakchi@gmail.com
An endophytic fungus Alternaria sp. SVJM05 was isolated from a medicinal plant, Indigofera ennaephylla Linn. and screened for its potential in anticancer production. The fungus was identified based on the morphology of the fungal culture and the characteristics of the spores. This fungus was grown in MID liquid medium and analyzed by chromatographically and spectrometrically for the presence of anticancer compound. The amount of anticancer compound produced by this fungus was quantified using high performance liquid chromatography (HPLC). The highest amount of anticancer compound production was recorded in the present study and further confirmative analyses were done by using TLC, UV, IR and LC-MS spectroscopy. The results designate that Alternaria sp. SVJM05 is an excellent candidate for anticancer compound production and thus, the fungus can serve as a potential material for fungus engineering to improve the production of anticancer compound. The effect of cytotoxicity of fungal anticancer compound isolated from Alternaria sp. SVJM05 was investigated by apoptosis method. The fungal anticancer compound extracted also showed a strong cytotoxic activity towards human cancer cells (HEP G-2) in vitro. The fungal extract was clearly showed the activity against the liver cancer in this study. The present findings suggest that Alternaria sp. SVJM05 could be an excellent alternative source for anticancer compound and may serve as a potential genetic engineered species for the enhanced production of anticancer compound.