36th Ann. Meeting of MSI & Nat. Seminar on Fungal Biodiversity & Bioprospecting….. Goa, India Oct., 29-30, 2009
POSTER SESSION 5
PS – 51
Herbal antimicrobials: A biological approach for purification of water
Awadhesh Kumar*, Rohit K. Mishra, Shalu Srivastava, Amit Kumar Tiwari, Kumar Pankaj Pandey, Ahsan Kamran, Rita Gupta*, Ameritesh Chandra Shukla* and Anupam Dikshit
Biological Product Laboratory, Dept. of Botany, University of Allahabad, Allahabad 211 002.
*Department of Horticulture Medicinal and Aromatic Plants, Mizoram University, Aizwal.
Water purification, is the process of removing bacteria, viruses and other biological contaminants from surface water or groundwater to make it safe and potable for human consumption. A large number of Indian populations residing in rural areas are dependent on the supply of untreated water, which is the root cause of their ailments. The 80% population of rural areas deprived of organized system of collecting, treating and supply of drinking water. The dug wells, hand pumps and step wells are the main sources of drinking water in rural areas. Somewhere the water ponds, lakes or rivers are used for domestic purpose. So that 90% of the rural diseases may be attributed to the polluted water. Unlike cities whereas fairly large population is using water filters based on ultraviolet radiation technology, ozonised water and RO system. The rural population is thriving on the contaminated water supply due to lack of financial resources and other pressing essentialities of life. Therefore, it is vital that with increased emphasis on augmenting the source of drinking water, efforts should also be made simultaneously for ensuring its quality. In rural areas the availability of plant materials used in the purification and its acceptability as environmentally safe has to be ensured. Essential oils, extracted from wide variety of plant species like Cinnamomum zeylanicum, Syzygium aromaticum, Eucalyptus globulus, Mentha piperita, Ocimum kilimandcharium, Vicia faba and Parkinsonia aculeata have potent antimicrobial activity for removing, inhibiting or killing the growth of various microorganism in drinking water.
PS – 52
Aeromycometric investigation in industrially polluted environment (ambattur) in Chennai
**S. Bhuvaneswari, N. Jawahar*, S. Vinodh*, S. Hariharan*, Ashok Livingstone Alex* and N. K. Udaya Prakash*
Research and Development, MARINA LABS, 40, Anna Nedum Pathai, Choolaimedu, Chennai 600 094.
*Department of Biotechnology, Veltech Hightech Dr. Rangarajan Dr. Sakunthala Engineering College,
Alamadhi Road, Avadi, Chennai 600 062.
*E-mail: marinalabs@yahoo.co.in
Ambattur Municipality is situated on the western side of Chennai city at a distance of about 20 km on Chennai - Thiruvallur High Road. Ambattur is fast developing as an urban center. Government of Tamil Nadu has established an Industrial Estate in Ambattur which has attracted reputed manufacturers like TVS, DUNLOP, T.I. CYCLES, etc, to set up industries. A number of housing colonies have sprung up in the area. The people are mainly employed in industrial establishments. The Municipality spreads over an area of 40.36 Sq.km., and the population of Ambattur as per 2001 Census is 3,08,764. The area is densely occupied different industries emanating physical and chemical pollutants within the area. People living on this location report suffering due to suffocation and chemical irritants. In this environment a study on presence of Atmospheric fungi was conducted. The study was conducted during the period of July – August. Altogether a total of 20 different spots were selected and atmospheric fungi were studied by exposing Potato Dextrose Agar through Andersen single stage sampler (Aerotech Inc, Arizona, USA). The exposed plates were brought in the laboratory and the growing colonies were analyzed. The isolated colonies are classified under 26 species among which Aspergillus was represented by maximum (14 spp.) number of species followed by Penicillium (5 spp.). The detailed report on this study will be discussed.
PS – 53
Prevalence and seasonal periodicity of airborne fungal spores in indoors and outdoors of a coastal village in Pondicherry region
B. K. Nayak*, R. Anandu, K. Usha and Anima Nanda**
Dept. of Plant Biology and Biotechnology, K. M. Centre for P. G. Studies (Autonomous), Pondicherry 605 008.
**Dept. of Biomedical Engineering, Sathyabama University, Rajiv Gandhi Salai, Chennai 600 119.
Studies on the prevalence of airborne fungal spores with their seasonal periodicity, in the indoors and outdoors of the houses in a coastal village of Pondicherry was carried out by implementing Petri plate sedimentation method from October 2008 to September 2009. Composition and concentration of fungal pores considerably varied from indoors to outdoors as well as from season to season. Outdoor air harbored maximum fungal spores (53%) in comparison to indoor air (47%). Occurrence of fungal species was predominated with more number of propagules during mid winter (December) and early rainy (July) periods in comparison to other months. In qualitative analysis, out of the total species recorded, Aspergillus was found with the highest frequency and had eleven members i.e., A. awamori, A. fumigatus, A. niger, A. flavus, A. flavipes, A. nidulans, A. ochraceous, A. japonicus, A. terreus, A. versicolor and A. wentii, but quantitatively, Penicillium was isolated highest in its contribution to total CFUs followed by Aspergillus. Out of the 33 isolated fungal taxa, Aspergillus fumigatus, A. awamori, A. niger, Rhizopus stolonifer and Alternaria alternata were the predominant aeroallergens, which cause different types of respiratory/lung diseases in atopic human beings. In seasonal periodicity, winter contributed the maximum spore load (41%) followed by rainy (33%), summer was found the least (26%) in harboring the spore mass in the indoors and outdoors in the village environment. Alternaria alternata, which is accounted as a human allergen for sporosis inducer and an agent for hay fever and other pathologies, was also intermittently recorded. In addition to the above aero allergenic fungi, a few plant pathogenic, saprophytic, field and storage fungi were also recorded during the study period.
PS – 54
Photodynamic control of Trichophyton mentagrophytes var. mentagrophytes by acriflavin, an acridine dye
S. Jenefar*, V. Balaji and P. Ebenezer
Centre for Advanced Studies in Botany, University of Madras, Chennai 600 025.
E-mail: jenesfashion@yahoo.co.in
Photodynamic therapy (PDT) is a treatment that involves light to activate a photosensitizing agent (photosensitizer) in the presence of oxygen. The exposure of the photosensitizer to light results in the formation of Reactive Oxygen Species, such as singlet oxygen and free radicals, causing localized photodamage and cell death. Clinically, this reaction is described as cytotoxic and vasculotoxic. The response of Trichophyton mentagrophytes to treatment with acriflavin, a common photodynamic dye has been studied in an attempt to apply the technique of PDT. Acriflavin, an acridine dye was found to have acted as a photosensitizer, exerting a photodynamic effect on the test fungus. Acriflavin was effective in controlling growth of the Trichophyton mentagrophytes in the presence of light and oxygen at a concentration of 100 µg/mL. Radial growth and biomass decreased in the presence of light and dye. Sporulation and spore germination were affected to a considerable extent. The optiumum pH and temperature for photodynamic control was found to be 7 and 35˚C respectively. The overall lipid, protein and sterol metabolism were found to have been affected by the photosensitizer. The syntheisis of protective enzymes, catalase and peroxidase also decreased with an increase in the concentration of the dye. Damage to the fungal membrane was minimal since the photosensitizer is a permeable dye. The response of Trichophyton mentagrophytes to treatment with acriflavin, and their action detected by antifungal assay.
PS – 55
Extraction, estimation and detoxification of aflatoxins present in storage mycoflora of animal feeds
K. P. Kannan*
Department of Biotechnology
Bannari Amman Institute of Technology, Sathyamangalm Erode District.
*E-mail: kp_kannan2001@yahoo.co.in
Aflatoxins are naturally occurring mycotoxins that are produced by many species of Aspergillus, most notably Aspergillus flavus and Aspergillus parasiticus. Aflatoxins are toxic and among the most carcinogenic substances known. After entering the body, aflatoxins are metabolized by the liver to a reactive intermediate, aflatoxin M1, an epoxide. The native habitat of Aspergillus is in soil, decaying vegetation, hay, and grains undergoing microbiological deterioration and it invades all types of organic substrates whenever conditions are favorable for its growth. Favorable conditions include high moisture content (at least 7%) and high temperature. Crops which are frequently affected include spices like chile pepper, black pepper, coriander, turmeric, ginger. At least 13 different types of aflatoxin are produced in nature. Aflatoxin B1 is considered the most toxic and is produced by both Aspergillus flavus and A .parasiticus.Aflatoxin G1 and G2 are produced exclusively by A. parasiticus. While the presence of Aspergillus in food products does not always indicate harmful levels of aflatoxin are also present, it does imply a significant risk in consumption of that product. In the present investigation animal feeds i.e. Cotton seeds, Different refinery cakes (sunflower cakes, groundnut cakes, and sesamum cakes) were collected from the market and were analyzed for the presence of storage mycoflora and the Aflatoxin concentration were employed. The results will be discussed in detail.
PS – 56
Bioadsorption of heavy metals by metal tolerant filamentous fungi
Peralam Yegneswaran Prakash*, Vaibhav Gupta**, Indira Bairy**
*Mycology Laboratory, Department of Microbiology, Kasturba Medical College.
**Division of Biotechnology, Manipal Life Sciences Centre, Manipal University, Karnataka.
The property of certain types of inactive, dead, microbial biomass to bind and concentrate heavy metals from dilute aqueous is referred to as bioremediation. Certain algae, fungi and bacteria have been employed to affect bioremediation in the event of oil spillages, sludge disposal and industrial waste cleanup in a large scale and involve the solid and liquid phase. Contaminated soils with toxic heavy metal pose a ecological threat and use of filamentous fungi for Bioremediation remains a choice owing to its robust nature and a potential of toxic metal recovery. To investigate bioabsorption properties of metal tolerant filamentous fungi isolated from metal contaminated soil sources by atomic absorption spectrophotometer. The study involved 3 phases; (i) Sampling for isolation of fungi from polluted soil sites (ii) Screening the heavy metal tolerance of the fungi to Copper, Lead, Iron and Cobalt using the Agar dilution and Spore germination assay (iii) Investigation of the bioadsorption properties of the metal tolerant fungi by atomic absorption spectrophotometer. Aspergillus niger, Aspergillus terreus, Absidia corymbifera, Paeciliomyces varioti and Trichoderma viridae were isolated from the polluted soil sites. In comparison to the 5 fungi studied; A. terreus was found to be tolerant to a relatively higher amount of Copper, Lead and Iron in the concentration of 35ppm, 25ppm and 200ppm respectively. The bioadsoprtion was influenced by physical parameters like temperature, pH and live versus dead mycelial biomass of Aspergillus terreus. The potential of Aspergillus terreus in bioremediation owing to their bioadsorption properties were found to be encouraging. Further studies by dose modulation effects on the genetic improvement of the strains could yield a promi
sing bioremediation agent to quench the increasing soil heavy metal toxicity making the planet a better place to live.
PS – 57
Growth control of the dermatophyte Trichophyton mentagrophytes var. mentagrophytes by a dual mode of action with Griseofulvin
D. Raynukaa*, V. Balaji V. and P. Ebenezer
Centre for Advanced Studies in Botany, University of Madras, Chennai 600 025.
*E-mail: raynukaa@gmail.com
Photodynamic therapy (PDT) involves light-sensitive photosensitizer, light, and molecular oxygen. After excitation with visible light, highly cytotoxic singlet oxygen and other reactive oxygen species (ROS) are generated by electron transfer. Singlet oxygen is a very reactive molecule that can damage many components of the target cell, resulting in cell death. Singlet oxygen can also destroy the photosensitizing agent itself preventing further action, a process referred to as photobleaching. Griseofulvin, an antifungal drug was found to have acted as a photosensitizer, exerting a photodynamic effect on the test fungus. The response of Trichophyton mentagrophytes to treatment with Griseofulvin, an antifungal drug, has been studied in an attempt to apply the technique of PDT. Griseofulvin was effective in controlling the growth of Trichophyton mentagrophytes in the presence of light and oxygen at a concentration of 100 µg/mL. Radial growth and biomass decreased in the presence of light and dye. Sporulation and spore germination were affected to an appreciable extent. The optiumum pH and temperature for photodynamic action of grieseofulvin were found to be 7 and 35˚C respectively. The overall lipid, protein and sterol metabolism were found to have been affected by the photosensitizer. The syntheisis of protective enzymes, catalase and peroxidase also decreased with an increase in the concentration of the dye. Damage to the fungal membrane appeared to be minimal since the photosensitizer is a permeable dye and the damage was endogenous. The response of Trichophyton mentagrophytes to treatment with grieseofulvin, and their action detected by antifungal assay.
PS – 58
X-ray induced mutation in Ganoderma species for higher anti thrombotic activity
P. Samuel Naveen* and M. Eyini
Research Centre in Botany, Thiagarajar College (Autonomous), Madurai 625009.
*E-mail: jeyini2005@yahoo.co.in
Thrombosis refers to the formation of a blood clot, thrombus within a vein which can partially or completely block the flow of the blood in the vein causing an embolism in the major organs and is life threatening. The mortality and morbidity resulting from thrombosis has been a major and increasing public health problem in India with a prevalence rate of 200 per 100,000 of the population per year. Thrombosis may be attributed to a sedentary lifestyle due to which disruption to blood flow occurs in the lower limbs and to an inappropriate diet that predisposes to acute thrombotic events. A regular diet containing ingredients with proven anti-thrombotic effect might be a convenient and effective way to prevent the disease. The use of Ganoderma lucidum as a dietary supplement has been documented in Chinese pharmacopoeia since approximately second century B.C. and it has been implicated against several diseases and physiological disorders like hypertension, diabetes and aging-related malfunctions. The present study investigated the effect of a physical mutagen, X–ray in increasing the antithrombotic activity of Ganoderma species in vitro. Physical mutation by X–Ray was induced by exposing PDA plates containing 6 days-old cultures of three pure cultures/wild isolates of Ganoderma lucidum (GL1, GL3, and GL5) and one pure culture of G. applanatum (GA2 IARI4655) for 0.8, 1.2, 1.6, 2.0 and 2.4 seconds at the rate of 40 mA / 75 KV from a X-ray machine (100 mA GE, INDIA). Based on mycelial morphology, growth rate and ligninase enzyme plate assays, eighty mutants were selected for the screening experiments. In the preliminary set of experiments, the PD broth aliquots of 14 days- old cultures of the four parent strains and their respective mutants were used to study the anti-thrombotic effect in whole blood. Among the parent strains, GL1 showed a significant increase in clotting time of 28sec after correcting for the dilution factor. The increase in clotting time of the other parent strains ranged between 21 sec (GA2),18.3(GL3) and 12sec (GL5) compared to the time required for clotting to occur under normal conditions (216sec). The mutants 2XLa4, 2.4XLa4, 2.4XLc4, 2.4XLc2, 1.6XLd4, 2.4XLd3, 1.6XAb3 and 2.4XAb1 showed significantly higher anti-thrombotic effect by extending the clotting time by 71 to 200 seconds compared to the parent strains at room temperature. These eight mutant strains were further subjected to coagulability tests, viz. Prothrombin Time (PT) and the Activated Partial Thromboplastin Time (APTT) using citrated plasma. The parent strains showed an increase in Prothrombin time which ranged between 19sec (GA2) and 49 sec (GL1) over that of the control (22sec) while the APTT values shown by the parents were extended by 6sec (GA2) and 29sec (GL3) over the control (25 sec) after allowing the effects of dilution. The pure culture of G. lucidum (GL1- IARI 1995) and its mutants (2XLa4, 2.4XLa4) showed the highest anti-thrombotic activity. PT & APTT duration of G. applanatum was less than that of the G. lucidum strains but its mutant, 2.4XAb1 showed significantly higher PT & APTT values than the latter. The mutants selected after the exposure time of 1.6, 2.4 and 2.0 seconds viz. 2XLc4, 2.4XLc2 (GL3 mutants) and1.6XLd4 (GL5 mutant).showed the maximum prothrombin time. The longest APTT was shown by 2XLc4, 2.4XLc2 (GL3 mutants) and 2.4XAb1, 1.6 XAb3 (GA2 mutants). PT and APTT results were more selective and confirmatory as some of the mutants of GL1 (2XLa4) and GL5 (2.4XLd3) which showed significant results in the screening tests using whole blood were found to be less efficient than their parents. The liquid growth media and the mycelia of the screened mutants thus represent a potential source for new therapeutic agents to treat thrombosis. However further characterization to study molecular level changes in the mutants are needed which may also pave the way for the production of new nutriceutical compounds for the treatment of thrombosis.
Tuesday, October 27, 2009
Technical Sessions-Abstracts-Oral session 5-OS-51-OS-54
36th Ann. Meeting of MSI & Nat. Seminar on Fungal Biodiversity & Bioprospecting….. Goa, India Oct., 29-30, 2009
ORAL SESSION 5
OS – 51
Biodegradation of Pentachlorophenol by white rot fungi isolated from forests of Western Ghats of Karnataka India.
Ch. Ramesh and Manohar G. Pattar*
Mycology Laboratory Department of Botany, Karnatak University, Dharwad 580 003 (Karnataka).
*E-mail: pgmanohar@gmail.com
Pentachlorophenol (PCP) is a major organic compound used in the wood preservation. White rot fungal strains screened from local sources such as wood trunks and twigs of higher plants. Five selected isolates of White Rot Fungi (WRF) Viz: Laetiporus cincinnatus, Trametes versicolor, Fomes fomentarius, Ganoderma aplanutun and Pleurotus ostreatus isolated from Sambrani and Dandeli forests of Western Ghats of Karnataka. The isolates were evaluated for the in-vitro biodegradation of PCP. The presence of extracellular laccase shows that these isolates are capable of degrading variety of xenobiotics including biodegradation of PCP. The maximum laccase activity (0.14 ± 003U) of F. fomentarius, P. ostreatus followed by T. versicolor study has shown significant efficiency on biodegradation of PCP which was quantitatively estimated and analyzed by High Performance Liquid Chromatography (HPLC). The static culture studies have been shown among all the isolates F. fomentarius (100%) and P. ostreatus (99.40±1.29%) degraded the PCP over 30 days of incubation.
OS – 52
L-asparaginase from fungi
T. S. Murali, Geetha Suvarna, P. M. Gopinath and K. Satyamoorthy
Division of Biotechnology, Manipal Life Sciences Centre, Manipal University,
Manipal 576 104 Karnataka.
Asparaginase (L-asparagine amido hydrolase, E.C. 3.5.1.1) is an anti-neoplastic agent and used in combination with other chemo-therapeutic agents in the treatment of acute leukemia. L-asparaginase (molecular weight-139,000) belongs to an amidase group that produces aspartic acid and ammonia by asparagine hydrolysis. L-asparaginase is produced by many bacteria, plants and animals such as E. coli, Erwinia sp., Candida utilis. Microbial asparaginases are being investigated for their therapeutic potential in treating acute lymphoblastic leukemia, and carcinomas. However, the bacterial enzyme causes few side effects such as liver damage, respiratory distress and hence there is a need for eukaryotic source of this enzyme. Though, fungi such as Aspergillus and Fusarium are known to produce asparaginase, very few studies have been carried out with regard to endophytic fungi. Endophytic fungi elicit several metabolites of medicinal importance and hence we screened endophytes isolated from medicinal plants such as Catharanthus roseus, Coleus sp. for their L-asparaginase production. The results were compared with L-asparaginase produced by filamentous fungi isolated from soil and water environments. Both qualitative and quantitative analyses of the enzyme were carried out. The results will be discussed in detail.
OS – 53
Marine derived fungi and their potential in metal bioremediation
Sarita Nazareth
Department of Microbiology, Goa University, Goa 403 206.
Marine-derived fungi comprise the obligate marine fungi and facultative species that are found in terrestrial ecosystems as well. These marine fungi have been discovered in different marine habitats exposed to various conditions, some of which may be extreme environments and the microorganisms adapted to withstand these harsh conditions. Pollution of our aquatic ecosystems by the influx of heavy metals has become a matter of concern over the last few decades. This toxic metal waste cannot be degraded by biological and chemical processes, nor can they be decomposed by in situ biological means and thus persist indefinitely in the environment, affecting organisms at the biochemical and cellular level: enzymes, nucleic acids, transport systems for essential nutrients and ions, displacement of essential ions from cellular sites, and disruption of cell and organ membrane integrity, eventually affecting community and population dynamics, thus causing severe damage to aquatic life, accumulating through the food chain and posing a grave threat to life. These deleterious effects and the emergence of more severe environmental protection laws have encouraged studies about removal and / or recovery of heavy metals from aqueous solutions. This can be achieved using biosorption or bioaccumulation which provides an attractive cost-effective, environment-friendly alternative to the physico-chemical method.The complex structure of microorganisms affords different means for metals to be taken up by the microbial cell, including ionic interactions and a passive sorption, not requiring live cells, or an energy-dependent, active uptake by live cells The fungi that predominate the estuarine waters, Goa, are observed to belong mainly to the genera of Aspergillus and Penicillium, with a few others of Cladosporium, Alternaria, Fusarium. Studies have shown that the fungi tolerate fairly high concentrations of heavy metals and react by developing a thickened cell wall perhaps due to increased chitin deposition, or a slow, dense mycelial aggregation as a means of protection and survival; they may also show morphological abnormalities such as swollen apical tips, or a constricted growth or an increase / decrease / absence / de novo synthesis of pigment production.
OS – 54
Onychomycosis - a potential threat to gardeners
Shaveta Jandial* and Geeta Sumbali
Department of Botany,
University of Jammu, Jammu 180 006
*E-mail: shavetajandial@yahoo.com
Onychomycosis means infection of nails caused by dermatophytes, yeasts and non-dermatophytic filamentous fungi. Previously researchers believed dermatophytes to be most commonly responsible for onychomycosis. But the spectrum of moulds capable of inducing onychomycosis is larger than commonly assumed, as many non-dermatophytic fungi such as Aspergillus spp., Fusarium spp., Onchocola spp., Scopulariopsis spp. etc have been recovered from nails. Farmers and gardeners are generally thought to be at higher risk of geophilic dermatomycoses because of their occupational contact with soil. Keeping this in view, a survey was undertaken to assess the pathogenic potential of dermatophytic/non-dermatophytic moulds from the toe and finger nails of 14 gardeners working in the botanical garden of University of Jammu. Visual observations showed that finger nails of only one gardener were affected. Frequent isolations from the nail clippings done on SDA (Sabouraud’s dextrose agar medium) yielded Fusarium equisetii as the causal agent. This is a new addition to the already existing list of non-dermatophytic agents of onychomycosis.
Myco-keratinophiles recovered from the soils of botanical garden of Jammu University also showed a preference for efficiently colonizing and utilizing nail keratin, thus predicting their pathogenic potential for onychomycosis.
ORAL SESSION 5
OS – 51
Biodegradation of Pentachlorophenol by white rot fungi isolated from forests of Western Ghats of Karnataka India.
Ch. Ramesh and Manohar G. Pattar*
Mycology Laboratory Department of Botany, Karnatak University, Dharwad 580 003 (Karnataka).
*E-mail: pgmanohar@gmail.com
Pentachlorophenol (PCP) is a major organic compound used in the wood preservation. White rot fungal strains screened from local sources such as wood trunks and twigs of higher plants. Five selected isolates of White Rot Fungi (WRF) Viz: Laetiporus cincinnatus, Trametes versicolor, Fomes fomentarius, Ganoderma aplanutun and Pleurotus ostreatus isolated from Sambrani and Dandeli forests of Western Ghats of Karnataka. The isolates were evaluated for the in-vitro biodegradation of PCP. The presence of extracellular laccase shows that these isolates are capable of degrading variety of xenobiotics including biodegradation of PCP. The maximum laccase activity (0.14 ± 003U) of F. fomentarius, P. ostreatus followed by T. versicolor study has shown significant efficiency on biodegradation of PCP which was quantitatively estimated and analyzed by High Performance Liquid Chromatography (HPLC). The static culture studies have been shown among all the isolates F. fomentarius (100%) and P. ostreatus (99.40±1.29%) degraded the PCP over 30 days of incubation.
OS – 52
L-asparaginase from fungi
T. S. Murali, Geetha Suvarna, P. M. Gopinath and K. Satyamoorthy
Division of Biotechnology, Manipal Life Sciences Centre, Manipal University,
Manipal 576 104 Karnataka.
Asparaginase (L-asparagine amido hydrolase, E.C. 3.5.1.1) is an anti-neoplastic agent and used in combination with other chemo-therapeutic agents in the treatment of acute leukemia. L-asparaginase (molecular weight-139,000) belongs to an amidase group that produces aspartic acid and ammonia by asparagine hydrolysis. L-asparaginase is produced by many bacteria, plants and animals such as E. coli, Erwinia sp., Candida utilis. Microbial asparaginases are being investigated for their therapeutic potential in treating acute lymphoblastic leukemia, and carcinomas. However, the bacterial enzyme causes few side effects such as liver damage, respiratory distress and hence there is a need for eukaryotic source of this enzyme. Though, fungi such as Aspergillus and Fusarium are known to produce asparaginase, very few studies have been carried out with regard to endophytic fungi. Endophytic fungi elicit several metabolites of medicinal importance and hence we screened endophytes isolated from medicinal plants such as Catharanthus roseus, Coleus sp. for their L-asparaginase production. The results were compared with L-asparaginase produced by filamentous fungi isolated from soil and water environments. Both qualitative and quantitative analyses of the enzyme were carried out. The results will be discussed in detail.
OS – 53
Marine derived fungi and their potential in metal bioremediation
Sarita Nazareth
Department of Microbiology, Goa University, Goa 403 206.
Marine-derived fungi comprise the obligate marine fungi and facultative species that are found in terrestrial ecosystems as well. These marine fungi have been discovered in different marine habitats exposed to various conditions, some of which may be extreme environments and the microorganisms adapted to withstand these harsh conditions. Pollution of our aquatic ecosystems by the influx of heavy metals has become a matter of concern over the last few decades. This toxic metal waste cannot be degraded by biological and chemical processes, nor can they be decomposed by in situ biological means and thus persist indefinitely in the environment, affecting organisms at the biochemical and cellular level: enzymes, nucleic acids, transport systems for essential nutrients and ions, displacement of essential ions from cellular sites, and disruption of cell and organ membrane integrity, eventually affecting community and population dynamics, thus causing severe damage to aquatic life, accumulating through the food chain and posing a grave threat to life. These deleterious effects and the emergence of more severe environmental protection laws have encouraged studies about removal and / or recovery of heavy metals from aqueous solutions. This can be achieved using biosorption or bioaccumulation which provides an attractive cost-effective, environment-friendly alternative to the physico-chemical method.The complex structure of microorganisms affords different means for metals to be taken up by the microbial cell, including ionic interactions and a passive sorption, not requiring live cells, or an energy-dependent, active uptake by live cells The fungi that predominate the estuarine waters, Goa, are observed to belong mainly to the genera of Aspergillus and Penicillium, with a few others of Cladosporium, Alternaria, Fusarium. Studies have shown that the fungi tolerate fairly high concentrations of heavy metals and react by developing a thickened cell wall perhaps due to increased chitin deposition, or a slow, dense mycelial aggregation as a means of protection and survival; they may also show morphological abnormalities such as swollen apical tips, or a constricted growth or an increase / decrease / absence / de novo synthesis of pigment production.
OS – 54
Onychomycosis - a potential threat to gardeners
Shaveta Jandial* and Geeta Sumbali
Department of Botany,
University of Jammu, Jammu 180 006
*E-mail: shavetajandial@yahoo.com
Onychomycosis means infection of nails caused by dermatophytes, yeasts and non-dermatophytic filamentous fungi. Previously researchers believed dermatophytes to be most commonly responsible for onychomycosis. But the spectrum of moulds capable of inducing onychomycosis is larger than commonly assumed, as many non-dermatophytic fungi such as Aspergillus spp., Fusarium spp., Onchocola spp., Scopulariopsis spp. etc have been recovered from nails. Farmers and gardeners are generally thought to be at higher risk of geophilic dermatomycoses because of their occupational contact with soil. Keeping this in view, a survey was undertaken to assess the pathogenic potential of dermatophytic/non-dermatophytic moulds from the toe and finger nails of 14 gardeners working in the botanical garden of University of Jammu. Visual observations showed that finger nails of only one gardener were affected. Frequent isolations from the nail clippings done on SDA (Sabouraud’s dextrose agar medium) yielded Fusarium equisetii as the causal agent. This is a new addition to the already existing list of non-dermatophytic agents of onychomycosis.
Myco-keratinophiles recovered from the soils of botanical garden of Jammu University also showed a preference for efficiently colonizing and utilizing nail keratin, thus predicting their pathogenic potential for onychomycosis.
Technical Sessions-Abstracts-Poster Session 4-PS-41-PS-54
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.
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.
Technical Sessions-Abstracts-Oral session 4-OS-41-OS-48
36th Ann. Meeting of MSI & Nat. Seminar on Fungal Biodiversity & Bioprospecting….. Goa, India Oct., 29-30, 2009
ORAL SESSION 4
OS – 41
Biosynthesis of Silver Nanoparticles Using Fungi
Anima Nanda*, Bijaya Kumar Nayak**
Department of Biomedical Engineering, Sathyabama University, Rajiv Gandhi Salai, Chennai 600 119.
**Department of Plant Biology & Biotechnology, K. M. Centre for P.G. Studies (Autonomous),
Airport Road, Pondicherry 605 008.
*E-mail: biju_nandu@yahoo.com
Nanotechnology is the science of creating or modifying materials at the atomic and molecular level to develop new or enhanced materials and products. Silver bio-nanoparticles (Ag-NPs) are known to possess bactericidal effects. The present study aims at development of extra-cellular biosynthesis of silver bio-nanoparticles using Aspergillus niger, Aspergillus flavus and Trichoderma harzianum and evaluated their antimicrobial activity against pathogenic bacteria. Ag-NPs will be synthesized by reducing Ag+ ions with the culture supernatants of Aspergillus niger, Aspergillus flavus and Trichoderma harzianum. The bio-reduction of the Ag+ ions in the solution assayed spectrophotometrically and allowed further confirmed by atomic force microscopy (AFM). The nano-bioparticles so obtained was evaluated for their antimicrobial activities against the target pathogen using standard methods of optimization. The most sensitive antimicrobial activity has been observed and zone of inhibition was recorded against various pathogens.
OS – 42
Production, Purification, Characterization and Biodiesel production by Extracellular lipases from oil seed-associated-fungi using Solid State Fermentation
V. Balaji* and P. Ebenezer
Centre for Advanced Studies in Botany, University of Madras, Chennai 600 025.
*E-mail: kvsbalaj@gmail.com
Oil-yielding seeds were collected from different places in the state of Tamil Nadu, India. In total, 1279 fungal isolates were recovered from the oil seeds. The fungal isolates were screened for their lipolytic activity. Out of 1279, only 40 isolates turned out to be promising interms of the enzyme yield onTween-20 Agar. These taxa were analysed stastically for the Colonization Frequency percentage (CF%), Diversity Indices, Shannon-Index H', Dominance Index, Simpson’s Index D, Species Richness (Chao) in oil seeds. The oil-yielding seeds exhibited rich fungal diversity. Quantification analysis of lipase production by these isolates revealed that Aspergillus niger (94 U/mL), Chalaropsis thielavioides (96 U/mL), Lasiodiplodia theobromae (108 U/mL), Phoma glomerata (95 U/mL) and Colletotrichum gloeosporioides (98 U/mL) produced appreciable quantities of the enzyme. Out of the above five good lipase producing isolates, Lasiodiplodia theobromae produced the maximum lipase units and therefore, the same isolate was chosen for further studies. The taxonomic identity of Lasiodiplodia theobromae was estabilished both morphometry and molecular methods. Lipase-producing ability was evaluated by Solid State Fermentation (SSF). Coconut oil cake appeared to be the best fermentation substrate for lipase production by L. theobromae VBE1 and hence the same was used through out the study. Optimization of lipase production by Response Surface Methodology (RSM) involved four factors― Central Composite Design involving 30 experiments was employed. On validating the predicted levels from the RSM, the enzyme production rose steeply. The enzyme so optimized was purified and characterized subsequently. Two distinct bands denoting two isoforms Lip A and Lip B appeared on the gels were separated by Q-Sepharose Column and Gel Filtration Chromatographic columns. The molecular mass of Lip A as determined on SDS-PAGE was 68 kDa and for Lip B it was 32 kDa. Purified lipase was used to generate Fatty Acid Methyl Esters (FAME/Biodiesel) from coconut and pogamia oils by both Lip A and NaOH catalytic action and were analysed by GC, GC-MS, IR, 1H NMR, 13C NMR. The other details will be presented in the paper.
OS – 43
Cardiovascular- protective and antimicrobial activity of a lichen species Usnea complanata
N. D. Mahadik, M. V. Morey and B. C. Behera
Agharkar Research Institute, G.G. Agarkar Road, Pune 411 004 (M.S.)
The purpose of the present study was to investigate the cardiovascular protective and antimicrobial properties of a lichen Usnea complanata. The biological active compounds the thallus of Usnea complanata was extracted in various solvents viz. methanol, ethanol, acetone, hexane, ethyl acetate and dimethyl sulfoxide (DMSO). As far as cardiovascular protective properties of U. complanata is concerned ethyl acetate extract showed maximum HMG-CoA reductase (HMGR) inhibitory activity up to 52.87% at 200μg/ml. Ethanol extract showed 46.37% inhibitory activity against angiotensin converting enzyme (ACE). Maximum fibrinolytic activity was obtained in methanol extract followed by hexane extract. Fibrinolytic activity showed by methanol extract at 100μg/ml was more than that of the activity obtained by standard plasmin at 10μg/ml. As far as antimicrobial activity of U. complanata is concerned, all extracts except the DMSO and hexane showed inhibitory activity against bacteria and fungi. Ethyl acetate extract was found to be the most efficient, since MIC90 was found in the range of (4.61μg/ml – 21.55μg/ml) which was low as compared to MIC90 of all other extract, against all bacterial cultures tested. Regarding the antifungal activity of extract of U. complanata, acetone and ethyl acetate extract inhibited all the tested fungi. The MIC ranged from 6.25 to 100μg/ml for ethyl acetate extract and was 12.5 to 100μg/ml for acetone extract.
OS – 44
Lichen metabolites developed in vitro and their antioxidative properties
Neeraj Verma* and B.C. Behera
Mycology & Plant Pathology Group,
Agharkar Research Institute, G.G. Agarkar Road, Pune 411 004 (M.S.).
*Email: neerajverma2310@gmail.com
Epidemiological studies have shown that free radicals have the potential to damage biological compounds and structures such as proteins, membrane lipids, DNA and carbohydrates and that such damages caused by these radicals are said to be involved in the etiology and pathogenesis of different diseases such as carcinogenesis, coronary heart disease, and many other health problems associated at advancing age. Antioxidants are considered as possible protective agents against oxidative damage. Therefore, there is a growing interest in the substances exhibiting antioxidant properties that are supplied to humans as food components or as specific pharmaceutical products. Recently, natural antioxidants have become one of the major areas of scientific research. The plant kingdom offers a wide range of natural antioxidants. However, little is known about the practical usefulness of most of them. Lichens are stable and self supporting symbioses between fungi (the mycobiont) and photoautotrophic algal partners (the photobiont). Lichen-forming fungi are potential sources of interesting novel metabolites known as “lichen substances”. These lichen metabolites are known to exert multiple pharmacological activities and properties such as antibiotic, antimycobacterial, antiviral, antiinflammatory, antioxidative, analgesic, antipyretic, antiproliferative and cytotoxic effects.
With this background information cell aggregates composed of mycobiont and photobiont derived from three lichen species, Arthothelium awasthii, Heterodermia podocarpa and Parmotrema tinctorum, were grown in Lilly-Barnett (LB) and Modified Bold’s Basal (MBB) media for one year under laboratory conditions. The results on growth rate, production of lichen substances barbatic acid, zeorin and lecanoric acid, and the antioxidant potential of extracts prepared with different solvents and of the partially purified substances are presented. The growth rate of the cell aggregates of the three species was found to be more or less similar in the LB and MBB media. The production of lichen substances by the three species was found to vary in both the media. The dried, soluble extract yield found from the different solvents was in the order of DMSO > methanol > acetone > ethyl acetate > chloroform > n-hexane. The antioxidant activities showed by barbatic acid, zeorin and lecanoric acid was highest at a concentration of 50µg/ml.
OS – 45
Bioactive polysaccharide isolated from pink oyster mushroom Pleurotus djamor var. roseus
N. Raaman*, R. Jegadeesh, L. Hariprasath, G. Rajasekhar Reddy, S. Archana and K. Balasubramanian
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
Higher fungi are abundant sources of a wide range of useful natural products and new products with interesting biological activities. Many species in polysaccharide, a big class of natural macromolecules, have been found to carry significant biological activities. It has been reported that polysaccharides derived from medicinal mushroom have activities of anti-infection, anti-agening, anti-radiation, immunomodulation and hepatoprocetive etc. All these activities are due to its contribution to enhance immune function of the human body. The presence study is focused to find out the medicinal properties of Pleurotus djamor var. roseus. The mushroom cultivated in Centre for Advanced Studies in Botany mushroom shed, freshly harvested mushroom were dried and make it a coarse powder. Water soluble glucans was isolated from basidiocarp of P.djamor var. roseus, by subsequent hot water extraction and ethanol precipitation. Obtained crude polysaccharide was treated with sevage reagent for removal of protein contaminations. The resulted partially purified polysaccharide was passing through DEAE cellulose Ion exchange chromatography and sapharose 6B gel permeation chromatography. The resulted FrA, fraction was characterized by analytical method. The spectroscopic analysis confirmed that purified fraction was a polysaccharide. Isolated polysaccharide was tested for in vitro antioxidant activity and its showed the IC50 of DPPH and hydroxyl radical scavenging activity 165 μg and 190 μg concentrations.
OS – 46
Optimization of medium and process parameters for the production of lipase from a newly isolated Aspergillus sp (RBD-01)
Satnam Singh Aulakh, N.Tejo Prakash*, Ranjana Prakash**
School of Chemistry and Biochemistry, Thapar University, Patiala.
*Department of Biotechnology and Environmental Sciences, Thapar University, Patiala.
**E-mail: rprakash@thapar.edu
Extracellular lipase production by Aspergillus sp. was monitored by modulating pH of the growth medium, ambient temperature for growth, source of nitrogen and percentage of carbon (virgin cottonseed oil). This strain was observed to be viable and produce lipase even up to 50% oil as a main carbon source. Maximum lipase activity of 21.8 U/ml was obtained with 50% (v/v) oil as main carbon source and peptone (0.5% w/v) as nitrogen source. The optimum pH and temperature for enzymatic activity was observed to be 7.5 and 35°C respectively. The observations are of significance due to limited reports on use 50% of oil as main carbon source and obtaining significant lipase activity of 21.8U/ml.
OS – 47
Evaluation the Probable mechanism of action of fungal bioactive compounds through bioinformatics
Shyamji Shukla* and A.K. Pandey
Mycology Research laboratory, Department of Biological Sciences, RDVV, Jabalpur (M.P.)
*Email: ji_shyam1@yahoo.com
Computer-based assessment of potential toxicity has become increasingly popular in recent years. Determination of toxicity of the drugs used in human welfare is still a big challenge for all the scientist working on discovery and development of drugs, due to its failure in certain instances. Interest in the use of Computer-based programs for toxicity prediction based on chemical structure is increasing due to the high cost and societal objections to animal testing, and the large numbers of candidate chemicals for screening. This challenge is global in its objective, but limited in its solution, to within local domains of chemicals acting according to similar mechanisms of action in the biological system. To predict the mechanisms of action of new fungal bioactive compoundsmycotoxins, some degree of generalization based on chemical structure must be possible through PASS (Prediction of activity structure for substances). The concept of the biological activity spectrum was introduced to describe the properties of biologically active substances. The PASS is a software product, use to predict pharmacological effects and biochemical mechanisms on the basis of the structural, formula of a compound. It may be efficiently used to find new targets for some ligands and, conversely, to reveal new ligands for some biological targets. Therefore, realizing the need and importance of this software, the present work was carried out with objective to predict the mechanism of action of fungal bioactive compounds obtained from several plant pathogenic fungi with the help of this tool.
OS – 48
Selection of potential isolates of Laccaria fraterna and Pisolithus albus on growth improvement of Acacia auriculiformis and A. mangium
V. Mohan* and P. Manokaran
Forest Pathology Lab., Forest Protection Division, Institute of Forest Genetics and Tree Breeding
Coimbatore 641 002.
In India, though a number of mycorrhizas have been collected, identified and described by various researchers but most of the work appears on endomycorrhizal fungi with agricultural, horticultural and few forestry crops. Limited reports are available on ectomycorrhizal association with economically important forestry species like Acacias, Eucalypts and other plantation crops. The host dependency on mycorrhizal fungi is common phenomenon in forest ecosystem but its efficiency is influenced by numerous biotic and abiotic factors particularly much depends upon the host plants and fungal isolates/strain. Therefore, there is a need for development of reliable and less expensive technique to inoculate nursery seedlings with suitable type of inoculum with selected ectomycorrhizal fungi for their better survival, establishment, growth and yield of the species on their transplantation to the new areas or fields. This may also provide an alternate use of costly fertilizers and help in afforestation programme and reclamation of wasteland particularly mined overburdens, sand dunes and other adverse sites. Acacia auriculiformis and A. mangium have been identified as the most promising tree species for both plantation forestry and reforestation in the tropics and also used for wood pulp, sawn timber and wood for particle board. In the present study, an investigation was made to select suitable type of ectomycorrhizal inoculum for growth enhancement of commercially important Acacia tree species in nursery. Different types of inocula such as basidiospore inoculum, vegetative mycelial vermiculite based inoculum and alginate bead inoculum were applied to seedlings of Acacia spp. in nursery. Data on growth parameters such as shoot & root lengths, leaf numbers, collar diameter, fresh and dry weights of shoot and roots of both inoculated (with different types of ECM fungal inocula) and uninoculated (control) seedlings of Acacia auriculiformis and A. mangium were recorded. The results the study indicated that basidiospore and vegetative mycelial inocula of L. fraterna and some of the isolates of P. albus inoculated seedlings had greater growth performance than uninoculated (control) seedlings. Morphological and anatomical features of roots of both inoculated and uninoculated (control) seedlings of Acacia species were also studied for assessing the persistence of the inoculant ECM fungi. The seedlings inoculated with vegetative mycelial inoculum and basidiospore inoculum of P. albus revealed more mycorrhizal tips in the roots of Acacia spp. in sterilized (autoclaved) potting medium as compared to unsterilized potting medium. Significance of the findings is discussed.
ORAL SESSION 4
OS – 41
Biosynthesis of Silver Nanoparticles Using Fungi
Anima Nanda*, Bijaya Kumar Nayak**
Department of Biomedical Engineering, Sathyabama University, Rajiv Gandhi Salai, Chennai 600 119.
**Department of Plant Biology & Biotechnology, K. M. Centre for P.G. Studies (Autonomous),
Airport Road, Pondicherry 605 008.
*E-mail: biju_nandu@yahoo.com
Nanotechnology is the science of creating or modifying materials at the atomic and molecular level to develop new or enhanced materials and products. Silver bio-nanoparticles (Ag-NPs) are known to possess bactericidal effects. The present study aims at development of extra-cellular biosynthesis of silver bio-nanoparticles using Aspergillus niger, Aspergillus flavus and Trichoderma harzianum and evaluated their antimicrobial activity against pathogenic bacteria. Ag-NPs will be synthesized by reducing Ag+ ions with the culture supernatants of Aspergillus niger, Aspergillus flavus and Trichoderma harzianum. The bio-reduction of the Ag+ ions in the solution assayed spectrophotometrically and allowed further confirmed by atomic force microscopy (AFM). The nano-bioparticles so obtained was evaluated for their antimicrobial activities against the target pathogen using standard methods of optimization. The most sensitive antimicrobial activity has been observed and zone of inhibition was recorded against various pathogens.
OS – 42
Production, Purification, Characterization and Biodiesel production by Extracellular lipases from oil seed-associated-fungi using Solid State Fermentation
V. Balaji* and P. Ebenezer
Centre for Advanced Studies in Botany, University of Madras, Chennai 600 025.
*E-mail: kvsbalaj@gmail.com
Oil-yielding seeds were collected from different places in the state of Tamil Nadu, India. In total, 1279 fungal isolates were recovered from the oil seeds. The fungal isolates were screened for their lipolytic activity. Out of 1279, only 40 isolates turned out to be promising interms of the enzyme yield onTween-20 Agar. These taxa were analysed stastically for the Colonization Frequency percentage (CF%), Diversity Indices, Shannon-Index H', Dominance Index, Simpson’s Index D, Species Richness (Chao) in oil seeds. The oil-yielding seeds exhibited rich fungal diversity. Quantification analysis of lipase production by these isolates revealed that Aspergillus niger (94 U/mL), Chalaropsis thielavioides (96 U/mL), Lasiodiplodia theobromae (108 U/mL), Phoma glomerata (95 U/mL) and Colletotrichum gloeosporioides (98 U/mL) produced appreciable quantities of the enzyme. Out of the above five good lipase producing isolates, Lasiodiplodia theobromae produced the maximum lipase units and therefore, the same isolate was chosen for further studies. The taxonomic identity of Lasiodiplodia theobromae was estabilished both morphometry and molecular methods. Lipase-producing ability was evaluated by Solid State Fermentation (SSF). Coconut oil cake appeared to be the best fermentation substrate for lipase production by L. theobromae VBE1 and hence the same was used through out the study. Optimization of lipase production by Response Surface Methodology (RSM) involved four factors― Central Composite Design involving 30 experiments was employed. On validating the predicted levels from the RSM, the enzyme production rose steeply. The enzyme so optimized was purified and characterized subsequently. Two distinct bands denoting two isoforms Lip A and Lip B appeared on the gels were separated by Q-Sepharose Column and Gel Filtration Chromatographic columns. The molecular mass of Lip A as determined on SDS-PAGE was 68 kDa and for Lip B it was 32 kDa. Purified lipase was used to generate Fatty Acid Methyl Esters (FAME/Biodiesel) from coconut and pogamia oils by both Lip A and NaOH catalytic action and were analysed by GC, GC-MS, IR, 1H NMR, 13C NMR. The other details will be presented in the paper.
OS – 43
Cardiovascular- protective and antimicrobial activity of a lichen species Usnea complanata
N. D. Mahadik, M. V. Morey and B. C. Behera
Agharkar Research Institute, G.G. Agarkar Road, Pune 411 004 (M.S.)
The purpose of the present study was to investigate the cardiovascular protective and antimicrobial properties of a lichen Usnea complanata. The biological active compounds the thallus of Usnea complanata was extracted in various solvents viz. methanol, ethanol, acetone, hexane, ethyl acetate and dimethyl sulfoxide (DMSO). As far as cardiovascular protective properties of U. complanata is concerned ethyl acetate extract showed maximum HMG-CoA reductase (HMGR) inhibitory activity up to 52.87% at 200μg/ml. Ethanol extract showed 46.37% inhibitory activity against angiotensin converting enzyme (ACE). Maximum fibrinolytic activity was obtained in methanol extract followed by hexane extract. Fibrinolytic activity showed by methanol extract at 100μg/ml was more than that of the activity obtained by standard plasmin at 10μg/ml. As far as antimicrobial activity of U. complanata is concerned, all extracts except the DMSO and hexane showed inhibitory activity against bacteria and fungi. Ethyl acetate extract was found to be the most efficient, since MIC90 was found in the range of (4.61μg/ml – 21.55μg/ml) which was low as compared to MIC90 of all other extract, against all bacterial cultures tested. Regarding the antifungal activity of extract of U. complanata, acetone and ethyl acetate extract inhibited all the tested fungi. The MIC ranged from 6.25 to 100μg/ml for ethyl acetate extract and was 12.5 to 100μg/ml for acetone extract.
OS – 44
Lichen metabolites developed in vitro and their antioxidative properties
Neeraj Verma* and B.C. Behera
Mycology & Plant Pathology Group,
Agharkar Research Institute, G.G. Agarkar Road, Pune 411 004 (M.S.).
*Email: neerajverma2310@gmail.com
Epidemiological studies have shown that free radicals have the potential to damage biological compounds and structures such as proteins, membrane lipids, DNA and carbohydrates and that such damages caused by these radicals are said to be involved in the etiology and pathogenesis of different diseases such as carcinogenesis, coronary heart disease, and many other health problems associated at advancing age. Antioxidants are considered as possible protective agents against oxidative damage. Therefore, there is a growing interest in the substances exhibiting antioxidant properties that are supplied to humans as food components or as specific pharmaceutical products. Recently, natural antioxidants have become one of the major areas of scientific research. The plant kingdom offers a wide range of natural antioxidants. However, little is known about the practical usefulness of most of them. Lichens are stable and self supporting symbioses between fungi (the mycobiont) and photoautotrophic algal partners (the photobiont). Lichen-forming fungi are potential sources of interesting novel metabolites known as “lichen substances”. These lichen metabolites are known to exert multiple pharmacological activities and properties such as antibiotic, antimycobacterial, antiviral, antiinflammatory, antioxidative, analgesic, antipyretic, antiproliferative and cytotoxic effects.
With this background information cell aggregates composed of mycobiont and photobiont derived from three lichen species, Arthothelium awasthii, Heterodermia podocarpa and Parmotrema tinctorum, were grown in Lilly-Barnett (LB) and Modified Bold’s Basal (MBB) media for one year under laboratory conditions. The results on growth rate, production of lichen substances barbatic acid, zeorin and lecanoric acid, and the antioxidant potential of extracts prepared with different solvents and of the partially purified substances are presented. The growth rate of the cell aggregates of the three species was found to be more or less similar in the LB and MBB media. The production of lichen substances by the three species was found to vary in both the media. The dried, soluble extract yield found from the different solvents was in the order of DMSO > methanol > acetone > ethyl acetate > chloroform > n-hexane. The antioxidant activities showed by barbatic acid, zeorin and lecanoric acid was highest at a concentration of 50µg/ml.
OS – 45
Bioactive polysaccharide isolated from pink oyster mushroom Pleurotus djamor var. roseus
N. Raaman*, R. Jegadeesh, L. Hariprasath, G. Rajasekhar Reddy, S. Archana and K. Balasubramanian
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
Higher fungi are abundant sources of a wide range of useful natural products and new products with interesting biological activities. Many species in polysaccharide, a big class of natural macromolecules, have been found to carry significant biological activities. It has been reported that polysaccharides derived from medicinal mushroom have activities of anti-infection, anti-agening, anti-radiation, immunomodulation and hepatoprocetive etc. All these activities are due to its contribution to enhance immune function of the human body. The presence study is focused to find out the medicinal properties of Pleurotus djamor var. roseus. The mushroom cultivated in Centre for Advanced Studies in Botany mushroom shed, freshly harvested mushroom were dried and make it a coarse powder. Water soluble glucans was isolated from basidiocarp of P.djamor var. roseus, by subsequent hot water extraction and ethanol precipitation. Obtained crude polysaccharide was treated with sevage reagent for removal of protein contaminations. The resulted partially purified polysaccharide was passing through DEAE cellulose Ion exchange chromatography and sapharose 6B gel permeation chromatography. The resulted FrA, fraction was characterized by analytical method. The spectroscopic analysis confirmed that purified fraction was a polysaccharide. Isolated polysaccharide was tested for in vitro antioxidant activity and its showed the IC50 of DPPH and hydroxyl radical scavenging activity 165 μg and 190 μg concentrations.
OS – 46
Optimization of medium and process parameters for the production of lipase from a newly isolated Aspergillus sp (RBD-01)
Satnam Singh Aulakh, N.Tejo Prakash*, Ranjana Prakash**
School of Chemistry and Biochemistry, Thapar University, Patiala.
*Department of Biotechnology and Environmental Sciences, Thapar University, Patiala.
**E-mail: rprakash@thapar.edu
Extracellular lipase production by Aspergillus sp. was monitored by modulating pH of the growth medium, ambient temperature for growth, source of nitrogen and percentage of carbon (virgin cottonseed oil). This strain was observed to be viable and produce lipase even up to 50% oil as a main carbon source. Maximum lipase activity of 21.8 U/ml was obtained with 50% (v/v) oil as main carbon source and peptone (0.5% w/v) as nitrogen source. The optimum pH and temperature for enzymatic activity was observed to be 7.5 and 35°C respectively. The observations are of significance due to limited reports on use 50% of oil as main carbon source and obtaining significant lipase activity of 21.8U/ml.
OS – 47
Evaluation the Probable mechanism of action of fungal bioactive compounds through bioinformatics
Shyamji Shukla* and A.K. Pandey
Mycology Research laboratory, Department of Biological Sciences, RDVV, Jabalpur (M.P.)
*Email: ji_shyam1@yahoo.com
Computer-based assessment of potential toxicity has become increasingly popular in recent years. Determination of toxicity of the drugs used in human welfare is still a big challenge for all the scientist working on discovery and development of drugs, due to its failure in certain instances. Interest in the use of Computer-based programs for toxicity prediction based on chemical structure is increasing due to the high cost and societal objections to animal testing, and the large numbers of candidate chemicals for screening. This challenge is global in its objective, but limited in its solution, to within local domains of chemicals acting according to similar mechanisms of action in the biological system. To predict the mechanisms of action of new fungal bioactive compoundsmycotoxins, some degree of generalization based on chemical structure must be possible through PASS (Prediction of activity structure for substances). The concept of the biological activity spectrum was introduced to describe the properties of biologically active substances. The PASS is a software product, use to predict pharmacological effects and biochemical mechanisms on the basis of the structural, formula of a compound. It may be efficiently used to find new targets for some ligands and, conversely, to reveal new ligands for some biological targets. Therefore, realizing the need and importance of this software, the present work was carried out with objective to predict the mechanism of action of fungal bioactive compounds obtained from several plant pathogenic fungi with the help of this tool.
OS – 48
Selection of potential isolates of Laccaria fraterna and Pisolithus albus on growth improvement of Acacia auriculiformis and A. mangium
V. Mohan* and P. Manokaran
Forest Pathology Lab., Forest Protection Division, Institute of Forest Genetics and Tree Breeding
Coimbatore 641 002.
In India, though a number of mycorrhizas have been collected, identified and described by various researchers but most of the work appears on endomycorrhizal fungi with agricultural, horticultural and few forestry crops. Limited reports are available on ectomycorrhizal association with economically important forestry species like Acacias, Eucalypts and other plantation crops. The host dependency on mycorrhizal fungi is common phenomenon in forest ecosystem but its efficiency is influenced by numerous biotic and abiotic factors particularly much depends upon the host plants and fungal isolates/strain. Therefore, there is a need for development of reliable and less expensive technique to inoculate nursery seedlings with suitable type of inoculum with selected ectomycorrhizal fungi for their better survival, establishment, growth and yield of the species on their transplantation to the new areas or fields. This may also provide an alternate use of costly fertilizers and help in afforestation programme and reclamation of wasteland particularly mined overburdens, sand dunes and other adverse sites. Acacia auriculiformis and A. mangium have been identified as the most promising tree species for both plantation forestry and reforestation in the tropics and also used for wood pulp, sawn timber and wood for particle board. In the present study, an investigation was made to select suitable type of ectomycorrhizal inoculum for growth enhancement of commercially important Acacia tree species in nursery. Different types of inocula such as basidiospore inoculum, vegetative mycelial vermiculite based inoculum and alginate bead inoculum were applied to seedlings of Acacia spp. in nursery. Data on growth parameters such as shoot & root lengths, leaf numbers, collar diameter, fresh and dry weights of shoot and roots of both inoculated (with different types of ECM fungal inocula) and uninoculated (control) seedlings of Acacia auriculiformis and A. mangium were recorded. The results the study indicated that basidiospore and vegetative mycelial inocula of L. fraterna and some of the isolates of P. albus inoculated seedlings had greater growth performance than uninoculated (control) seedlings. Morphological and anatomical features of roots of both inoculated and uninoculated (control) seedlings of Acacia species were also studied for assessing the persistence of the inoculant ECM fungi. The seedlings inoculated with vegetative mycelial inoculum and basidiospore inoculum of P. albus revealed more mycorrhizal tips in the roots of Acacia spp. in sterilized (autoclaved) potting medium as compared to unsterilized potting medium. Significance of the findings is discussed.
Technical sessions-Abstracts-Poster Session 3-PS-31-PS-40
36th Ann. Meeting of MSI & Nat. Seminar on Fungal Biodiversity & Bioprospecting….. Goa, India Oct., 29-30, 2009
POSTER SESSION 3
PS – 31
Exploration of starch loaded habitats & samples for isolation of industrially useful amylolytic yeasts
M. Albuquerque and Nandkumar Kamat
Department of Botany, Goa University, Goa 403 206.
Yeasts represent a group of microbes widely distributed in natural ecosystems & significant for their role in food fermentations & enzyme industry. There is relatively scanty knowledge of tropical yeast diversity in natural & fermented media.
Amylolytic yeasts produce amylases that find application in the clarification of fruit juices, stain remover in detergents, preparation of glucose syrup & monosodium glutamate in food processing units, clarification of beer in breweries, as digestive tablets, prevents layering of chocolate syrup & in Desizing textiles.
The screening survey constitutes the first approach in selecting yeast strains exploitable as enzyme producers from various habitats such as tuber cultivating farms, plant storage organs e.g. Potato, sweet potato, cassava.; flour deposits from flour mills, local bread sellers, traditional bakeries. All these habitats can harbor potentially wild amylolytic yeast strains.
Samples were collected from starch loaded habitats in North & South Goa, documented, transported & processed, resulting in the isolation of 119 yeast isolates, which after enzymological screening 15 hyper amylase producers were obtained, which will form a culture bank of wild type amylolytic strains that would be of tremendous use to the food industry.
PS – 32
Bioefficacy of Lactic acid bacteria in the management of Aspergillus japonicus and Rhizopus stolonifer rot of Apricots (Prunus armeniaca L.)
J. Bhadwal and Y.P. Sharma*
Department of Botany, University of Jammu, Jammu, India 180 006.
*E-mail: yashdbm3@yahoo.co.in
Aspergillus japonicus and Rhizopus stolonifer are the most notorious mycopathogens of fresh apricots inciting serious decay and rots. Keeping this in view, an attempt was made to control these two rots using Lactic acid bacteria, an ecofriendly and biosafe biocontrol agent. In the present investigation, lyophilized cells of Lactic acid bacteria were obtained, rehydrated and used in three varying concentrations (1x1012 cfu/ml, 1x1010 cfu/ml and 1x109 cfu/ml) for both pre- and post-infection treatments. It was found that all the three concentrations of Lactic acid bacteria that were used in both pre- and post-infection treatments of fresh apricots inoculated with A. japonicus showed a rot control percentage ranging from 68.81% ± 0.87 to 97.04% ± 0.57 and 37% ± 0.94 to 69.49% ± 1.59 respectively. On the other hand, pre- and post-infection treatments of apricot fruits inoculated with R. stolonifer gave a rot control percentage in the range of 65.18%
± 0.76 to 93.87% ± 0.88 and 0% respectively, indicating clearly relatively more efficacy of pre-infection treatment than post-infection treatment and positive correlation between the concentration of the biocontrol agent and the percent control of fruit rot. It was, thus, concluded that LAB were efficacious and could be used for increasing the shelf-life of apricot fruits.
PS – 33
Phytochemical composition and in vitro antifungal activity of Citrus limon
Mala Majumdar, C. G. Sangeetha, S. Karthik, B. Seema Pai , Krupa B. Paleja and Shwetha H. Gowda
Department of Biotechnology, Centre for Post-Graduate Studies, Jain University,
#18/3, 9th Main, III Block, Jayanagar, Bangalore 560 011.
Citrus limon (lemon) has natural plant chemicals (phytochemicals) which has medicinal value. Lemon also shows some antimicrobial activity. To evaluate the scientific basis of the plant, the pulp, leaves and rind extracts were evaluated against Candida albicans, Aspergillus niger, Fusarium sp .and Penicillium sp. The antifungal activity of the concentrated extracts was evaluated by determination of the diameter of the zone of inhibition against fungi using agar well diffusion method. Aqueous extract of pulp, leaves and rind showed inhibition against C. albicans whereas ethanol extract of pulp inhibited the growth of Fusarium sp .and Penicillium sp. Standard procedures were used to identify the bioactive principles like alkaloids, tannins, saponins, terpenoids, flavonoids and cardiac glycoside. Qualitative analysis revealed the presence of saponins, terpenoids and cardiac glycosides in pulp and tannins, terpenoids, flavonoids and cardiac glycosides in lemon rind. Lemon pulp showed the presence of tannins, terpenoids and cardiac glycosides whereas rind showed the presence of all the phytochemicals except alkaloids and tannins. Quantitative estimation of crude phytochemical constituents showed highest percentage of flavonoids in pulp and saponins in pulp and rind. Antifungal activity of the phytochemical tested showed that flavonoids extracted from pulp showed inhibition against C. albicans whereas saponins from rind and pulp also inhibited C. albicans.
PS – 34
Evaluation of phytochemical composition and antifungal activity of Aegle marmelos
Mala Majumdar, C. G. Sangeetha, Krupa B. Paleja, Shwetha H. Gowda, S. Karthik and B. Seema Pai
Department of Biotechnology, Centre for Post-Graduate Studies, Jain University,
#18/3, 9th Main, III Block, Jayanagar, Bangalore 560 011.
An assessment of phytochemical composition and antifungal activity of aqueous and ethanol extract of bark, leaves, pulp and rind of Aegle marmelos (Bael) against Candida albicans, Aspergillus niger, Fusarium sp .and Penicillium sp. was carried out. The antifungal activities of extracts were compared with standard antibiotics. The antifungal activity of both the extracts was performed by agar well diffusion method. Maximum inhibition of A. niger was recorded by aqueous extract of the leaves and pulp showed inhibition against C. albicans and rind extract showed inhibition against all the tested fungi. The ethanol extract did not show any inhibition against any of the tested fungi. Phytochemical tests were carried out on the aqueous extract using standard procedures to identify the bioactive principles like alkaloids, tannins, saponins, terpenoids, flavonoids and cardiac glycoside. Pulp showed the presence of all the above constituents whereas except tannins all the other active principles were present in bark. Flavonoids and cardiac glycoside were not present in leaves and rind did not show the presence of alkaloid. Quantitative estimation of the percentage of crude phytochemical constituents showed highest percentage of alkaloid in bark and flavonoid in rind. Antifungal activity of the phytochemicals tested showed that the flavanoids extracted from pulp and rind showed inhibition against Candida albicans. Alkaloids and flavonoids from bark extract, saponins from rind extract and alkaloid from bark extract showed inhibition against Penicillium sp. The study suggests that the rind and bark extracts has active principles which could be harnessed as an effective phytomedicine
PS – 35
Biological control of post harvest diseases of brinjal
Reeny Mary Zacharia
Rice Research Station, Moncompu, Alappuzha district, Kerala.
The fungi associated with the spoilage of brinjal fruits were studied. Aspergillus niger and Trichothecium roseum were occasionally present, while fungi like Alternaria solani, Fusarium solani, Colletotrichum gloeosporioides, Botrytis cinerea, Penicillium sp., Rhizopus niger, Curvularia lunata and Botryodiplodia theobromae caused damage through the year. Pythium vexans, Phytophthora palmivora, Pythium aphanidermatum and Rhizoctonia solani were seasonal in appearance and confined to rainy periods. 26 phylloplane fungi were isolated from the leaf surface of brinjal plants. The type of mycoparasitsm between the pathogen and the phylloplane fungi was studied. The pathogens viz., F. solani, A. solani were overgrown by the phylloplane fungi viz., T.viride, A.niger, and A.flavus. Homogenous intermingling was noticed with Phoma sp. and A. solani. Cessation of growth at line of growth was shown by P.palmivora with both pathogen, while a clear zone of inhibition was noticed with F. solani and A.terreus. The mode of action was studied for T.viride alone. Penetration and coiling of hyphae was noticed with F. solani, while penetration and disintegration was common with A. solani. In Trichoderma treated fruits eventhough symptoms were visible by fourth day, it caused only 48 percent damage by 12th day, while in control samples of F.solani infected fruits symptoms were observed by 12th day and complete rotting by 12th day. The study thus highlighted the possibility of utilizing Trichoderma as a biocontrol agent against the major diseases of brinjal.
PS – 36
Occurrence and distribution of keratinophilic fungi in soils of Khardung La, Ladakh (India)
Sandeep Kotwal* and Geeta Sumbali
Department of Botany, University of Jammu, Jammu 180 006.
*E-mail: s.kotwal@yahoo.com
Keratinophilic fungi are an ecologically important group of microorganisms, which cycle keratin that is one of the most abundant and highly stable animal proteins on the earth. This unique group of fungi consists of vigorous and self- sufficient saprophytes as long as environmental conditions are favourable. However, they are opportunists and may become parasitic by accident. After becoming pathogenic, they are able to survive and multiply at host’s body temperature, causing further infection by invading fresh keratinized tissues. It is the only type of fungal infection that is known to be of contagious type.
Realizing the significance of this unique group of fungi, research work was undertaken to study the occurrence and distribution of keratinophilic fungi from the unexplored soils of Khardung La (Ladakh), which is the highest motorable pass of India. It is about 17,582 feet above sea-level, geographically diverse region with cold climate, thick snow cover and comparatively less activity of humans and animals. Soil samples were collected from this area and keratinophiles were recovered by using keratin-bait technique.
Examination of the soil samples showed that 80% of them were positive for keratinophilic fungi. A total of seven keratinophilic fungal species were recovered and all of them produced only the imperfect state on Sabouraud’s dextrose agar (SDA) medium. The recovered fungal species consisted of Aspergillus flavus, A. sydowii, A. ustus, Beauveria bassiana, Fusarium oxysporum, F. sporotrichioides and F. moniliforme var. subglutinans. Among these, Aspergillus flavus, Beauveria bassiana and Fusarium oxysporum are known to be potential pathogenic fungi. During the study, Beauveria bassiana was found to be the most dominant keratinophilic species with 40% frequency occurrence; followed in decreasing order by Aspergillus flavus (30%), Fusarium oxysporum (30%), A. sydowii (20%), A. ustus (20%), F. sporotrichioides (20%) and F. moniliforme var. subglutinans (10%).
Low occurrence and distribution of keratinophilic species from Khardung La soils clearly indicates that harsh environmental conditions and meagre presence of keratinous substrates affect the growth, development and diffusion of keratinophilic fungi.
PS – 37
Study on the efficacy of the vegetative and spore inoculum of Pisolithus albus on the Casurina junghiniana
R. Shenbhagaraman* and V.Kaviyarasan
Centre for advanced studies in Botany, University of Madras, Guindy campus Chennai 600 025.
*E-mail: shenbhagaraman@rediffmail.com, shenbhagaraman@yahoo.co.in
The fruiting bodies of ectomycorrhizal fungus Pisolithus albus were collected from Eucalyptus plantations in around Chennai. The fungus was isolated and maintained over MMN medium. The mycelial growth was optimized and grown over the sterile vermiculite in order to use as vegetative inoculum. Spores were collected from matured fruit bodies were used as spore inoculum. The vegetative inoculum and spore inoculum were applied to one month old seedlings of Casurina junghuniana grown in sterile sand: red soil mixer in the ratio of 2: 1. The seedlings were maintained in the glass house for a period of one month and then transferred to the field. Ten seedlings were sacrificed from each group at the end of the fourth, six, eighth month and were studied for biomass, shoot height, root height, percentage of mycorrhization, starch, total chlorophyll, total carbohydrates, total protein, total nitrogen and micronutrients. The vegetative inoculum enhanced more growth than the spore inoculum. However the seedlings treated with spore inoculum is better than control. The results are discussed in detail.
PS – 38
Studies on characterisation of mycotoxigenic fungi from oil seed cakes and animal feeds
G. L. Sujatha and Ramalingappa
Department of studies in Microbiology, Kuvempu University, Shivagangothri Campus,
Davangere 577 002, Karnataka.
Mycotoxigenic fungi are the potent contaminants of storage foods and feed. The major contaminants are Aspergillus, Penicillium, Fusarium, most species of Aspergillus and Penicillum are able to grow on a wide range of organic substrate. Most of the feed and or feed ingredients are tested for the presence of toxigenic fungi in India. The oil seed Cakes and animal feed samples from Davangere were mycologically investigated in terms of overall fungal diversity and toxicological potential of isolated fungi. Totally 10 samples, 3 samples are oil seed Cakes and all other are animal feeds were randomly collected at intervals from new packets of feed samples in the market in Davangere and brought to laboratory and samples named as S1, S2, S3, S4, S5, S6, S7, S8, S9 and S10 respectively. From these samples mycotoxigenic fungi were isolated by Serial dilution method and direct plate method using different mycological media. Totally 13 genera of fungi was isolated. Aspergillus and Penicillium was identified upto species level. A.flavus detected rapidly on Aspergillus differential medium (ADM) and its toxigenicity tested by ammonia Vapour test. For Penicillium which produces cyclopiazonic acid or other alkaloids detected by Ehrlich test. Aflatoxins from A.flavus and A.parasiticus was purified and separated by Analytical Thin layer chromatographic method. Identified as AFB1, AFB2 & AFG and compared with standard AFB2. Cyclopiazonic acid from P. commune was also purified and identified by characteristic bluish-violet Pigment. By this the incidence of contamination of fungi in oil seed Cakes and animal feeds can be determined.
PS – 39
Laboratory Evaluation of the Entomopathogenic fungi, Aspergillus niger and Metarrhizium anisopliae for Controlling Selected pests
K. N. Varalakshmi and B. S. Kumudini
Department of Biotechnology, Centre for Post Graduate Studies, Jain University, #18/3, 9th Main, 3rd Block, Jayanagar, Bangalore 560 011.
Continuous use of chemicals at high rates against key insects led to pest resurgence resulting from development of resistance and destruction of natural enemies. Entomopathogenic fungi which are very virulent have been used in recent days to develop a logical approach. Application of integrated pest management greatly decreases the use of conventional pesticides, while crop yields remain high. In an attempt to identify such entomopathogenic fungi a study was conducted. In this study fungi were isolated from adult cockroaches, house fly, worms from cauliflower and cabbage from different locations. After proper identification of the fungi, infectivity trials were conducted on Sitophilus oryzae (rice weevil) and eggs of Periplanata americana (cockroach). Different fungi that were isolated were identified as Aspergillus niger, A. flavus, Trichoderma sp. and Penicillium sp. Infectivity trials with A. niger spores showed that by day 8 percentage mortality was 96+0.4 whereas, in control boxes percentage mortality was only 10+0.3. Results of the experiment showing the hatching rate (%) of P. americana eggs sprayed with A. niger spores was very encouraging. In the experimental boxes the percent hatching was greatly affected by A. niger infection. By 7 weeks after infection with A. niger spores, the hating rate was only 6.7% whereas in the control eggs of P. americana the hatching rate is 66.7%. Results of experiment showing mortality (%) of P. americana young ones sprayed with Metarrhizium anisopliae spores showed 100% mortality by the end of day 9. Further field infectivity trials will be carried out for evaluating the feasibility of using these fungi for the biological control of pests of economic importance.
PS – 40
Keratinophilic fungi of domestic animals at Gulbarga Region
G. M. Vidyasagar* and Narayan Hosmani
Medicinal Plants and Microbiology Laboratory, Department of P G Studies and Research in Botany, Gulbarga University, Gulbarga 585 106, Karnataka.
*E-mail:gmvidyasagar@rediffmail.com
Domestic animals play an important role in the transmission of dermatophytes and other potentially pathogenic fungi. Cow, ox, buffalo, sheep and goat are the common domestic animals in Gulbarga region and are very close to human life. Fifty hair samples from each animal were collected in sterilized polythene bags from different places of Gulbarga and incubated them on moistened sterile soil for six weeks at room temperature. The fungi grown on hair samples were transferred on to Sabouraud’s dextrose agar medium and identified with the help of literature. A total of 32 fungal species were identified. Of which, 10 were dermatophytes and 22 were other related ones. Ox hairs yielded maximum of 21 keratinophilic fungi, followed by Cow (20), goat (18), sheep (17) and buffalo (16). Maximum numbers of dermatophytes were isolated from cow and ox hairs (7). This was followed by buffalo (5), goat (4) and sheep hairs (3). Microsporium gypseum was recorded on ox, buffalo and cow hairs, while Trichophyton mentagrophytes recorded only on cow and ox hairs. No incidence of these two dermatophytes was observed on sheep and goat hairs. The occurrence of dermatophytes correlates with the relation of man with animals.
POSTER SESSION 3
PS – 31
Exploration of starch loaded habitats & samples for isolation of industrially useful amylolytic yeasts
M. Albuquerque and Nandkumar Kamat
Department of Botany, Goa University, Goa 403 206.
Yeasts represent a group of microbes widely distributed in natural ecosystems & significant for their role in food fermentations & enzyme industry. There is relatively scanty knowledge of tropical yeast diversity in natural & fermented media.
Amylolytic yeasts produce amylases that find application in the clarification of fruit juices, stain remover in detergents, preparation of glucose syrup & monosodium glutamate in food processing units, clarification of beer in breweries, as digestive tablets, prevents layering of chocolate syrup & in Desizing textiles.
The screening survey constitutes the first approach in selecting yeast strains exploitable as enzyme producers from various habitats such as tuber cultivating farms, plant storage organs e.g. Potato, sweet potato, cassava.; flour deposits from flour mills, local bread sellers, traditional bakeries. All these habitats can harbor potentially wild amylolytic yeast strains.
Samples were collected from starch loaded habitats in North & South Goa, documented, transported & processed, resulting in the isolation of 119 yeast isolates, which after enzymological screening 15 hyper amylase producers were obtained, which will form a culture bank of wild type amylolytic strains that would be of tremendous use to the food industry.
PS – 32
Bioefficacy of Lactic acid bacteria in the management of Aspergillus japonicus and Rhizopus stolonifer rot of Apricots (Prunus armeniaca L.)
J. Bhadwal and Y.P. Sharma*
Department of Botany, University of Jammu, Jammu, India 180 006.
*E-mail: yashdbm3@yahoo.co.in
Aspergillus japonicus and Rhizopus stolonifer are the most notorious mycopathogens of fresh apricots inciting serious decay and rots. Keeping this in view, an attempt was made to control these two rots using Lactic acid bacteria, an ecofriendly and biosafe biocontrol agent. In the present investigation, lyophilized cells of Lactic acid bacteria were obtained, rehydrated and used in three varying concentrations (1x1012 cfu/ml, 1x1010 cfu/ml and 1x109 cfu/ml) for both pre- and post-infection treatments. It was found that all the three concentrations of Lactic acid bacteria that were used in both pre- and post-infection treatments of fresh apricots inoculated with A. japonicus showed a rot control percentage ranging from 68.81% ± 0.87 to 97.04% ± 0.57 and 37% ± 0.94 to 69.49% ± 1.59 respectively. On the other hand, pre- and post-infection treatments of apricot fruits inoculated with R. stolonifer gave a rot control percentage in the range of 65.18%
± 0.76 to 93.87% ± 0.88 and 0% respectively, indicating clearly relatively more efficacy of pre-infection treatment than post-infection treatment and positive correlation between the concentration of the biocontrol agent and the percent control of fruit rot. It was, thus, concluded that LAB were efficacious and could be used for increasing the shelf-life of apricot fruits.
PS – 33
Phytochemical composition and in vitro antifungal activity of Citrus limon
Mala Majumdar, C. G. Sangeetha, S. Karthik, B. Seema Pai , Krupa B. Paleja and Shwetha H. Gowda
Department of Biotechnology, Centre for Post-Graduate Studies, Jain University,
#18/3, 9th Main, III Block, Jayanagar, Bangalore 560 011.
Citrus limon (lemon) has natural plant chemicals (phytochemicals) which has medicinal value. Lemon also shows some antimicrobial activity. To evaluate the scientific basis of the plant, the pulp, leaves and rind extracts were evaluated against Candida albicans, Aspergillus niger, Fusarium sp .and Penicillium sp. The antifungal activity of the concentrated extracts was evaluated by determination of the diameter of the zone of inhibition against fungi using agar well diffusion method. Aqueous extract of pulp, leaves and rind showed inhibition against C. albicans whereas ethanol extract of pulp inhibited the growth of Fusarium sp .and Penicillium sp. Standard procedures were used to identify the bioactive principles like alkaloids, tannins, saponins, terpenoids, flavonoids and cardiac glycoside. Qualitative analysis revealed the presence of saponins, terpenoids and cardiac glycosides in pulp and tannins, terpenoids, flavonoids and cardiac glycosides in lemon rind. Lemon pulp showed the presence of tannins, terpenoids and cardiac glycosides whereas rind showed the presence of all the phytochemicals except alkaloids and tannins. Quantitative estimation of crude phytochemical constituents showed highest percentage of flavonoids in pulp and saponins in pulp and rind. Antifungal activity of the phytochemical tested showed that flavonoids extracted from pulp showed inhibition against C. albicans whereas saponins from rind and pulp also inhibited C. albicans.
PS – 34
Evaluation of phytochemical composition and antifungal activity of Aegle marmelos
Mala Majumdar, C. G. Sangeetha, Krupa B. Paleja, Shwetha H. Gowda, S. Karthik and B. Seema Pai
Department of Biotechnology, Centre for Post-Graduate Studies, Jain University,
#18/3, 9th Main, III Block, Jayanagar, Bangalore 560 011.
An assessment of phytochemical composition and antifungal activity of aqueous and ethanol extract of bark, leaves, pulp and rind of Aegle marmelos (Bael) against Candida albicans, Aspergillus niger, Fusarium sp .and Penicillium sp. was carried out. The antifungal activities of extracts were compared with standard antibiotics. The antifungal activity of both the extracts was performed by agar well diffusion method. Maximum inhibition of A. niger was recorded by aqueous extract of the leaves and pulp showed inhibition against C. albicans and rind extract showed inhibition against all the tested fungi. The ethanol extract did not show any inhibition against any of the tested fungi. Phytochemical tests were carried out on the aqueous extract using standard procedures to identify the bioactive principles like alkaloids, tannins, saponins, terpenoids, flavonoids and cardiac glycoside. Pulp showed the presence of all the above constituents whereas except tannins all the other active principles were present in bark. Flavonoids and cardiac glycoside were not present in leaves and rind did not show the presence of alkaloid. Quantitative estimation of the percentage of crude phytochemical constituents showed highest percentage of alkaloid in bark and flavonoid in rind. Antifungal activity of the phytochemicals tested showed that the flavanoids extracted from pulp and rind showed inhibition against Candida albicans. Alkaloids and flavonoids from bark extract, saponins from rind extract and alkaloid from bark extract showed inhibition against Penicillium sp. The study suggests that the rind and bark extracts has active principles which could be harnessed as an effective phytomedicine
PS – 35
Biological control of post harvest diseases of brinjal
Reeny Mary Zacharia
Rice Research Station, Moncompu, Alappuzha district, Kerala.
The fungi associated with the spoilage of brinjal fruits were studied. Aspergillus niger and Trichothecium roseum were occasionally present, while fungi like Alternaria solani, Fusarium solani, Colletotrichum gloeosporioides, Botrytis cinerea, Penicillium sp., Rhizopus niger, Curvularia lunata and Botryodiplodia theobromae caused damage through the year. Pythium vexans, Phytophthora palmivora, Pythium aphanidermatum and Rhizoctonia solani were seasonal in appearance and confined to rainy periods. 26 phylloplane fungi were isolated from the leaf surface of brinjal plants. The type of mycoparasitsm between the pathogen and the phylloplane fungi was studied. The pathogens viz., F. solani, A. solani were overgrown by the phylloplane fungi viz., T.viride, A.niger, and A.flavus. Homogenous intermingling was noticed with Phoma sp. and A. solani. Cessation of growth at line of growth was shown by P.palmivora with both pathogen, while a clear zone of inhibition was noticed with F. solani and A.terreus. The mode of action was studied for T.viride alone. Penetration and coiling of hyphae was noticed with F. solani, while penetration and disintegration was common with A. solani. In Trichoderma treated fruits eventhough symptoms were visible by fourth day, it caused only 48 percent damage by 12th day, while in control samples of F.solani infected fruits symptoms were observed by 12th day and complete rotting by 12th day. The study thus highlighted the possibility of utilizing Trichoderma as a biocontrol agent against the major diseases of brinjal.
PS – 36
Occurrence and distribution of keratinophilic fungi in soils of Khardung La, Ladakh (India)
Sandeep Kotwal* and Geeta Sumbali
Department of Botany, University of Jammu, Jammu 180 006.
*E-mail: s.kotwal@yahoo.com
Keratinophilic fungi are an ecologically important group of microorganisms, which cycle keratin that is one of the most abundant and highly stable animal proteins on the earth. This unique group of fungi consists of vigorous and self- sufficient saprophytes as long as environmental conditions are favourable. However, they are opportunists and may become parasitic by accident. After becoming pathogenic, they are able to survive and multiply at host’s body temperature, causing further infection by invading fresh keratinized tissues. It is the only type of fungal infection that is known to be of contagious type.
Realizing the significance of this unique group of fungi, research work was undertaken to study the occurrence and distribution of keratinophilic fungi from the unexplored soils of Khardung La (Ladakh), which is the highest motorable pass of India. It is about 17,582 feet above sea-level, geographically diverse region with cold climate, thick snow cover and comparatively less activity of humans and animals. Soil samples were collected from this area and keratinophiles were recovered by using keratin-bait technique.
Examination of the soil samples showed that 80% of them were positive for keratinophilic fungi. A total of seven keratinophilic fungal species were recovered and all of them produced only the imperfect state on Sabouraud’s dextrose agar (SDA) medium. The recovered fungal species consisted of Aspergillus flavus, A. sydowii, A. ustus, Beauveria bassiana, Fusarium oxysporum, F. sporotrichioides and F. moniliforme var. subglutinans. Among these, Aspergillus flavus, Beauveria bassiana and Fusarium oxysporum are known to be potential pathogenic fungi. During the study, Beauveria bassiana was found to be the most dominant keratinophilic species with 40% frequency occurrence; followed in decreasing order by Aspergillus flavus (30%), Fusarium oxysporum (30%), A. sydowii (20%), A. ustus (20%), F. sporotrichioides (20%) and F. moniliforme var. subglutinans (10%).
Low occurrence and distribution of keratinophilic species from Khardung La soils clearly indicates that harsh environmental conditions and meagre presence of keratinous substrates affect the growth, development and diffusion of keratinophilic fungi.
PS – 37
Study on the efficacy of the vegetative and spore inoculum of Pisolithus albus on the Casurina junghiniana
R. Shenbhagaraman* and V.Kaviyarasan
Centre for advanced studies in Botany, University of Madras, Guindy campus Chennai 600 025.
*E-mail: shenbhagaraman@rediffmail.com, shenbhagaraman@yahoo.co.in
The fruiting bodies of ectomycorrhizal fungus Pisolithus albus were collected from Eucalyptus plantations in around Chennai. The fungus was isolated and maintained over MMN medium. The mycelial growth was optimized and grown over the sterile vermiculite in order to use as vegetative inoculum. Spores were collected from matured fruit bodies were used as spore inoculum. The vegetative inoculum and spore inoculum were applied to one month old seedlings of Casurina junghuniana grown in sterile sand: red soil mixer in the ratio of 2: 1. The seedlings were maintained in the glass house for a period of one month and then transferred to the field. Ten seedlings were sacrificed from each group at the end of the fourth, six, eighth month and were studied for biomass, shoot height, root height, percentage of mycorrhization, starch, total chlorophyll, total carbohydrates, total protein, total nitrogen and micronutrients. The vegetative inoculum enhanced more growth than the spore inoculum. However the seedlings treated with spore inoculum is better than control. The results are discussed in detail.
PS – 38
Studies on characterisation of mycotoxigenic fungi from oil seed cakes and animal feeds
G. L. Sujatha and Ramalingappa
Department of studies in Microbiology, Kuvempu University, Shivagangothri Campus,
Davangere 577 002, Karnataka.
Mycotoxigenic fungi are the potent contaminants of storage foods and feed. The major contaminants are Aspergillus, Penicillium, Fusarium, most species of Aspergillus and Penicillum are able to grow on a wide range of organic substrate. Most of the feed and or feed ingredients are tested for the presence of toxigenic fungi in India. The oil seed Cakes and animal feed samples from Davangere were mycologically investigated in terms of overall fungal diversity and toxicological potential of isolated fungi. Totally 10 samples, 3 samples are oil seed Cakes and all other are animal feeds were randomly collected at intervals from new packets of feed samples in the market in Davangere and brought to laboratory and samples named as S1, S2, S3, S4, S5, S6, S7, S8, S9 and S10 respectively. From these samples mycotoxigenic fungi were isolated by Serial dilution method and direct plate method using different mycological media. Totally 13 genera of fungi was isolated. Aspergillus and Penicillium was identified upto species level. A.flavus detected rapidly on Aspergillus differential medium (ADM) and its toxigenicity tested by ammonia Vapour test. For Penicillium which produces cyclopiazonic acid or other alkaloids detected by Ehrlich test. Aflatoxins from A.flavus and A.parasiticus was purified and separated by Analytical Thin layer chromatographic method. Identified as AFB1, AFB2 & AFG and compared with standard AFB2. Cyclopiazonic acid from P. commune was also purified and identified by characteristic bluish-violet Pigment. By this the incidence of contamination of fungi in oil seed Cakes and animal feeds can be determined.
PS – 39
Laboratory Evaluation of the Entomopathogenic fungi, Aspergillus niger and Metarrhizium anisopliae for Controlling Selected pests
K. N. Varalakshmi and B. S. Kumudini
Department of Biotechnology, Centre for Post Graduate Studies, Jain University, #18/3, 9th Main, 3rd Block, Jayanagar, Bangalore 560 011.
Continuous use of chemicals at high rates against key insects led to pest resurgence resulting from development of resistance and destruction of natural enemies. Entomopathogenic fungi which are very virulent have been used in recent days to develop a logical approach. Application of integrated pest management greatly decreases the use of conventional pesticides, while crop yields remain high. In an attempt to identify such entomopathogenic fungi a study was conducted. In this study fungi were isolated from adult cockroaches, house fly, worms from cauliflower and cabbage from different locations. After proper identification of the fungi, infectivity trials were conducted on Sitophilus oryzae (rice weevil) and eggs of Periplanata americana (cockroach). Different fungi that were isolated were identified as Aspergillus niger, A. flavus, Trichoderma sp. and Penicillium sp. Infectivity trials with A. niger spores showed that by day 8 percentage mortality was 96+0.4 whereas, in control boxes percentage mortality was only 10+0.3. Results of the experiment showing the hatching rate (%) of P. americana eggs sprayed with A. niger spores was very encouraging. In the experimental boxes the percent hatching was greatly affected by A. niger infection. By 7 weeks after infection with A. niger spores, the hating rate was only 6.7% whereas in the control eggs of P. americana the hatching rate is 66.7%. Results of experiment showing mortality (%) of P. americana young ones sprayed with Metarrhizium anisopliae spores showed 100% mortality by the end of day 9. Further field infectivity trials will be carried out for evaluating the feasibility of using these fungi for the biological control of pests of economic importance.
PS – 40
Keratinophilic fungi of domestic animals at Gulbarga Region
G. M. Vidyasagar* and Narayan Hosmani
Medicinal Plants and Microbiology Laboratory, Department of P G Studies and Research in Botany, Gulbarga University, Gulbarga 585 106, Karnataka.
*E-mail:gmvidyasagar@rediffmail.com
Domestic animals play an important role in the transmission of dermatophytes and other potentially pathogenic fungi. Cow, ox, buffalo, sheep and goat are the common domestic animals in Gulbarga region and are very close to human life. Fifty hair samples from each animal were collected in sterilized polythene bags from different places of Gulbarga and incubated them on moistened sterile soil for six weeks at room temperature. The fungi grown on hair samples were transferred on to Sabouraud’s dextrose agar medium and identified with the help of literature. A total of 32 fungal species were identified. Of which, 10 were dermatophytes and 22 were other related ones. Ox hairs yielded maximum of 21 keratinophilic fungi, followed by Cow (20), goat (18), sheep (17) and buffalo (16). Maximum numbers of dermatophytes were isolated from cow and ox hairs (7). This was followed by buffalo (5), goat (4) and sheep hairs (3). Microsporium gypseum was recorded on ox, buffalo and cow hairs, while Trichophyton mentagrophytes recorded only on cow and ox hairs. No incidence of these two dermatophytes was observed on sheep and goat hairs. The occurrence of dermatophytes correlates with the relation of man with animals.
Technical sessions-Abstracts-Oral Session 3-OS-31-OS-35
36th Ann. Meeting of MSI & Nat. Seminar on Fungal Biodiversity & Bioprospecting….. Goa, India Oct., 29-30, 2009
ORAL SESSION 3
OS – 31
Growth Response of Solanum nigrum L. to Arbuscular mycorrhizal fungi and Plant growth promoting microorganisms
K. V. Chethan Kumar, Sukesh* and Chandrashekar K.R*
Department of Microbiology, Post graduate study centre, MSRCASC, Bangalore.
*Department of Applied Botany, Mangalore University, Mangalagangothri, Mangalore.
Pot experiments were conducted to know the influence of inoculation with Arbuscular mycorrhizal fungi (AM fungi) and the Plant growth promoting microorganisms (PGPRMs). Efficacy of ten Arbuscular mycorrhizal fungi on Solanum nigrum (Medicinal Plant) was studied. Plants grown in the presence of AM fungi generally showed an increase in plant growth and biomass over those grown in the absence of AM fungal inoculation. It was observed that Glomus mosseae was the best AM symbiont to enhance the plant biomass. This best AM symbiont was selected to study the response of Solanum nigrum to the inoculation in combination with different PGPRMs (Trichoderma viridae, Aspergillus awamorii, Azotobacter chroococcum and Psuedomonas fluorescence). The plant height, number of leaves per plant, number of branches per plant, Plant dry and fresh matter, Plant P content and N content were significantly higher in plants inoculated in combination with Glomus mosseae and PGPRMs. A higher root colonization and phosphatase activity was also observed in the plants inoculated with Glomus mosseae alone and in combination. This study clearly brings out that inoculation with Glomus mosseae + PGPRMs not only improve the growth, biomass yield, P nutrition of Solanum nigrum, but also help in sustainable agricultural practices in the cultivation of Solanum nigrum.
OS – 32
Production, purification and characterization of an extracellular lipase from Aspergillus japonicus isolated from Ropalidia marginata paper nest
A. Jayaprakash* and P. Ebenezer
Centre for Advanced Studies in Botany, University of Madras, Guindy campus, Chennai 600 025.
*E-mail: aruljaypee@gmail.com
Lipases (EC 3.1.1.3) hydrolyze triglycerides to fatty acids and glycerol. They are widely distributed in nature. Although lipases have been found in many species of animals, plants, bacteria, yeast, and fungi, the enzymes from microorganisms are most interesting because of their potential applications in various industries such as food, dairy, pharmaceutical, detergents, textile,biodiesel, and cosmetic industries, and in synthesis of fine chemicals, agrochemicals, and new polymeric materials. Fungi are important enzyme producers since they produce enzymes extracellularly. The present investigation is aimed at the isolation of paper nest-inhabiting fungi of Ropalidia marginata and screening the same for extracellular lipase activity, identification of promising lipolytic fungal isolates and optimization of enzyme production of the isolates by response Surface Methodology (RSM). Further, purification and characterization of the extracellular lipase by Ion exchange and gel filtration chromatography.
OS – 33
Yeast Killer toxins elaborated by the anamorphic basidiomycetous yeast Cryptococcus neoformans towards various ascomycetous yeast genera
Peralam Yegneswaran Prakash, Taronish Dubash*, Sugandhi Rao**
Medical Mycology Laboratory, Department of Microbiology, Kasturba Medical College,
*Division of Biotechnology, Manipal Life Sciences Centre, Manipal University, Karnataka.
**Department of Microbiology and Immunology, Kasturba Medical College International Centre, Manipal University, Karnataka.
Yeast cells liberate lethal polypeptides toxins designated as killer proteins or killer toxins to which they themselves are immune. The killer yeasts produce toxins as either proteins or glycoproteins, and these toxins are fatal to other yeast strains. Not all yeasts strains are sensitive to killer toxins and yeasts are currently classified as killer strains, sensitive strains, unaffected strains and competitive yeast strains. In recent years several new killer yeast strains have been reported but not yet well classified. The activity of theses toxins is analogous to the activities of bacteriocins in bacterial strains. To isolate Cryptococcus neoformans from clinical and ecological sources and check for killer toxin elaborated by them and their interaction with the pathogenic and non-pathogenic ascomycetous yeast genera. The Killer yeast activity was estimated using the calorimetric measuring of cell growth using the Thiazole blue tetrazolium bromide (MTT) Assay where in the reduction takes place only when mitochondrial reductase enzymes are active and therefore conversion can be directly related to the number of viable cells. In addition the plate count agar (PCA) Assay using YEPG agar to determine the viable counts and Methylene Blue (MB) agar diffusion assay were performed. Data analyses were performed using SPSS 10.0 Software. Ecological isolates of Cryptococcus neoformans were found to elaborate the Killer phenotypes in comparison to the Clinical isolates (P<0.05). The Killer activity were documented against test strains of Pichia , Brettanomyces, Saccharomyces, Candida. The existence of killer phenotype in a relatively higher number in the ecological source may serve in the elimination of competitors, either literal killing or a severe reduction in the fitness of sensitive strains, through blocking one or a few specific vital biochemical reactions in their metabolic pathways or punching a hole in the cell membrane. Whatever actual form it takes, toxin production is a clear case of interference competition a type of ecological interaction differing from resource competition in that it involves a direct active suppression of the rival population by means other than just exhausting the common limiting resources.
OS – 34
Mycoparasitic Penicillium on Aspergillus niger causing pomegranate fruit rot
Rahul Sharma and S.K. Singh
National Facility for Culture Collection of Fungi, Agharkar Research Institute,
G.G. Agarkar Road, Pune 411004, MS.
Recently a Pomegranate fruit rot (post harvest) was encountered caused by a member of Aspergillus niger complex which currently comprises of nine species. During isolation of the fungus certain grayish conidial heads were seen in the centre of the fungal colonization, which on close observation with stereomicroscope appeared Penicillium like conidial apparatus which eventually was confirmed by microscopy and the two species were separated in pure form. Penicillium species are known to parasitize Aspergillus conidial heads (Horn & Peterson, 2008) but the phenomenon is quite uncommon. In our case, whether this association is strictly parasitic in nature is still a matter of speculation since no substantial Penicillium mycelium was seen growing inside Aspergillus niger hyphae. Molecular characterization was done to identify the two species involved and the present communication puts more light on this association between two most versatile mitosporic genera of Plectomycetes.
OS – 35
Fungal Endophytes: A Potential Source of Anticancer Compounds
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.
*E-mail: sunil.deshmukh@piramal.com
Endophytic fungi are microorganisms that colonize living, internal tissue of plants without causing any immediate, overt, negative effects or external symptoms. Recently, a great deal of interest has been generated by discovery of remarkable pharmacological agents from endophytic fungi and also recognized as potential source of bioactive secondary metabolites with a high level of structural diversity . These fungi are known to produce anticancer compound viz. Taxol, Vincristine, and Camptothecine which were originally obtained from the host plant. In the pre
sentation, the emphasis will be given to both endophytic fungi and their chemical diversity. Some emphasis will also be given to anticancer compounds produced by this group of fungi.The need of an extensive survey of this group of fungi from unexplored areas and exploitation of their ability to produce anticancer compounds, and culture collection of this group of fungi will also be highlighted.
ORAL SESSION 3
OS – 31
Growth Response of Solanum nigrum L. to Arbuscular mycorrhizal fungi and Plant growth promoting microorganisms
K. V. Chethan Kumar, Sukesh* and Chandrashekar K.R*
Department of Microbiology, Post graduate study centre, MSRCASC, Bangalore.
*Department of Applied Botany, Mangalore University, Mangalagangothri, Mangalore.
Pot experiments were conducted to know the influence of inoculation with Arbuscular mycorrhizal fungi (AM fungi) and the Plant growth promoting microorganisms (PGPRMs). Efficacy of ten Arbuscular mycorrhizal fungi on Solanum nigrum (Medicinal Plant) was studied. Plants grown in the presence of AM fungi generally showed an increase in plant growth and biomass over those grown in the absence of AM fungal inoculation. It was observed that Glomus mosseae was the best AM symbiont to enhance the plant biomass. This best AM symbiont was selected to study the response of Solanum nigrum to the inoculation in combination with different PGPRMs (Trichoderma viridae, Aspergillus awamorii, Azotobacter chroococcum and Psuedomonas fluorescence). The plant height, number of leaves per plant, number of branches per plant, Plant dry and fresh matter, Plant P content and N content were significantly higher in plants inoculated in combination with Glomus mosseae and PGPRMs. A higher root colonization and phosphatase activity was also observed in the plants inoculated with Glomus mosseae alone and in combination. This study clearly brings out that inoculation with Glomus mosseae + PGPRMs not only improve the growth, biomass yield, P nutrition of Solanum nigrum, but also help in sustainable agricultural practices in the cultivation of Solanum nigrum.
OS – 32
Production, purification and characterization of an extracellular lipase from Aspergillus japonicus isolated from Ropalidia marginata paper nest
A. Jayaprakash* and P. Ebenezer
Centre for Advanced Studies in Botany, University of Madras, Guindy campus, Chennai 600 025.
*E-mail: aruljaypee@gmail.com
Lipases (EC 3.1.1.3) hydrolyze triglycerides to fatty acids and glycerol. They are widely distributed in nature. Although lipases have been found in many species of animals, plants, bacteria, yeast, and fungi, the enzymes from microorganisms are most interesting because of their potential applications in various industries such as food, dairy, pharmaceutical, detergents, textile,biodiesel, and cosmetic industries, and in synthesis of fine chemicals, agrochemicals, and new polymeric materials. Fungi are important enzyme producers since they produce enzymes extracellularly. The present investigation is aimed at the isolation of paper nest-inhabiting fungi of Ropalidia marginata and screening the same for extracellular lipase activity, identification of promising lipolytic fungal isolates and optimization of enzyme production of the isolates by response Surface Methodology (RSM). Further, purification and characterization of the extracellular lipase by Ion exchange and gel filtration chromatography.
OS – 33
Yeast Killer toxins elaborated by the anamorphic basidiomycetous yeast Cryptococcus neoformans towards various ascomycetous yeast genera
Peralam Yegneswaran Prakash, Taronish Dubash*, Sugandhi Rao**
Medical Mycology Laboratory, Department of Microbiology, Kasturba Medical College,
*Division of Biotechnology, Manipal Life Sciences Centre, Manipal University, Karnataka.
**Department of Microbiology and Immunology, Kasturba Medical College International Centre, Manipal University, Karnataka.
Yeast cells liberate lethal polypeptides toxins designated as killer proteins or killer toxins to which they themselves are immune. The killer yeasts produce toxins as either proteins or glycoproteins, and these toxins are fatal to other yeast strains. Not all yeasts strains are sensitive to killer toxins and yeasts are currently classified as killer strains, sensitive strains, unaffected strains and competitive yeast strains. In recent years several new killer yeast strains have been reported but not yet well classified. The activity of theses toxins is analogous to the activities of bacteriocins in bacterial strains. To isolate Cryptococcus neoformans from clinical and ecological sources and check for killer toxin elaborated by them and their interaction with the pathogenic and non-pathogenic ascomycetous yeast genera. The Killer yeast activity was estimated using the calorimetric measuring of cell growth using the Thiazole blue tetrazolium bromide (MTT) Assay where in the reduction takes place only when mitochondrial reductase enzymes are active and therefore conversion can be directly related to the number of viable cells. In addition the plate count agar (PCA) Assay using YEPG agar to determine the viable counts and Methylene Blue (MB) agar diffusion assay were performed. Data analyses were performed using SPSS 10.0 Software. Ecological isolates of Cryptococcus neoformans were found to elaborate the Killer phenotypes in comparison to the Clinical isolates (P<0.05). The Killer activity were documented against test strains of Pichia , Brettanomyces, Saccharomyces, Candida. The existence of killer phenotype in a relatively higher number in the ecological source may serve in the elimination of competitors, either literal killing or a severe reduction in the fitness of sensitive strains, through blocking one or a few specific vital biochemical reactions in their metabolic pathways or punching a hole in the cell membrane. Whatever actual form it takes, toxin production is a clear case of interference competition a type of ecological interaction differing from resource competition in that it involves a direct active suppression of the rival population by means other than just exhausting the common limiting resources.
OS – 34
Mycoparasitic Penicillium on Aspergillus niger causing pomegranate fruit rot
Rahul Sharma and S.K. Singh
National Facility for Culture Collection of Fungi, Agharkar Research Institute,
G.G. Agarkar Road, Pune 411004, MS.
Recently a Pomegranate fruit rot (post harvest) was encountered caused by a member of Aspergillus niger complex which currently comprises of nine species. During isolation of the fungus certain grayish conidial heads were seen in the centre of the fungal colonization, which on close observation with stereomicroscope appeared Penicillium like conidial apparatus which eventually was confirmed by microscopy and the two species were separated in pure form. Penicillium species are known to parasitize Aspergillus conidial heads (Horn & Peterson, 2008) but the phenomenon is quite uncommon. In our case, whether this association is strictly parasitic in nature is still a matter of speculation since no substantial Penicillium mycelium was seen growing inside Aspergillus niger hyphae. Molecular characterization was done to identify the two species involved and the present communication puts more light on this association between two most versatile mitosporic genera of Plectomycetes.
OS – 35
Fungal Endophytes: A Potential Source of Anticancer Compounds
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.
*E-mail: sunil.deshmukh@piramal.com
Endophytic fungi are microorganisms that colonize living, internal tissue of plants without causing any immediate, overt, negative effects or external symptoms. Recently, a great deal of interest has been generated by discovery of remarkable pharmacological agents from endophytic fungi and also recognized as potential source of bioactive secondary metabolites with a high level of structural diversity . These fungi are known to produce anticancer compound viz. Taxol, Vincristine, and Camptothecine which were originally obtained from the host plant. In the pre
sentation, the emphasis will be given to both endophytic fungi and their chemical diversity. Some emphasis will also be given to anticancer compounds produced by this group of fungi.The need of an extensive survey of this group of fungi from unexplored areas and exploitation of their ability to produce anticancer compounds, and culture collection of this group of fungi will also be highlighted.
Technical Sessions-abstracts-Poster Session 2-PS-21-PS-33
36th Ann. Meeting of MSI & Nat. Seminar on Fungal Biodiversity & Bioprospecting….. Goa, India Oct., 29-30, 2009
POSTER SESSION 2
PS – 21
Isolation, identification and purification of endophytic fungi from medicinal plants for the enumeration of extracellular enzymes
A. Amrita, J. Swetha, P. Sindhu, and K. P. Kannan*
Department of Biotechnology, Bannari Amman Institute of technology
Satyamangalam, Erode district 638 401
*E-mail: kp_kannan2001@yahoo.co.in
Endophytes are microorganisms, which inhabit in healthy living plant tissues for all or part of their life cycle without causing apparent harmful symptoms to the host. In resent years, much research has been focused on the bioactivities of endophytic fungi. This interest is due to the possibility of these microorganisms producing pharmacologically active substances with biotechnological potential such as antitumor agents (Taxol), antifungal agents (quercine), besides producing factors of plant growth, toxins and enzymes, including some microorganisms being used as biological controllers of many diseases and plagues. In the present investigation four medicinal plants viz., Adathoda vasica, Costus ingneus, Coleus aromaticus, and Lawsonia inerims were examined for the presence of endophytic fungi. Altogether 400 segments (approximately1 cm2) of 50 segments each from leaf and stem tissues of respective plants were screened for the enumeration of the endophytic fungi using the modified surface sterilization techniques. A total of 17 different species of endophytic fungi were recorded. Among them 9 belongs to Hyphomycetes, 7 belongs to Coelomycetes and one belongs to Xylariales. From the endophytic fungi recorded, 10 dominant endophytic fungi viz., Cladosporium cladosporioides, Curvularia brachyspora, C.verruciformis, Drechslera hawaiiensis, Colletotrichum carssipes, Colletotrichum falctum, Colletotrichum gleosporioides, Lasiodiplodia theobromae, Phyllosticta Sp. and Xylariales species were tested for their ability to produce extracellular enzymes i.e., Amylase, Cellulase, Laccase, Lipase, Protease and Tyrosinase activity by the qualitative assays, majority of the entophytic fungi showed the positive results. The results will be discussed in detail.
PS – 22
Screening of endophytic fungi from the medicinal plants for the production of Chitoson
A. Amrita, P. Sindhu, J. Swetha, and K. P. Kannan*
Department of Biotechnology, Bannari Amman Institute of Technology, Satyamangalam,
Erode district 638 401.
*E-mail: kp_kannan2001@yahoo.co.in
Endophytic fungi that live inside the tissues of living plants are under-explored group of microorganisms. Recently they have received considerable attention after they were found to protect their host against insect pests, pathogens and even domestic herbivores. Medicinal plants are reported to harbour endophytes which in turn provide protection to their host from infectious agents and also provide adaptability to survive in adverse environmental conditions. It is therefore important to determine the endophytic diversity of medicinal plants. In the present study four medicinal plants were screened for the presence of Endophytic fungi and were screened for the production of chitin and Chitoson. Industrially, Chitosan is derived from the chemical deacetylation of chitin, using strong alkali, a waste product of the crustacean exoskeletons obtained after industrial processing of seafood. This process is subject to seasonal, limited supply and it pollutes the environment, due to the large amount of waste of concentrated alkaline solution. An alternative to solve these problems is the fungal production of Chitosan, whose main advantage is that it is environmentally friendly. Therefore, physicochemical properties and yields of chitosan isolated directly from a fungus may be optimised by controlling fermentation and processing parameters. The development of applications for chitosan has expanded rapidly in recent years. Chitosan, being polycationic, nontoxic, biodegradable as well as antimicrobial, has many applications especially in the agriculture, food and pharmaceutical industries. It has been used in enzyme immobilization, wastewater treatment, as food additive and anticholesterolemic, for wound healing, and in pharmaceuticals in several drug delivery systems. The results will be discussed in detail.
PS – 23
Partial purification of exracellular enzyme cellulase from Trichoderma viride and T. reesei isolated from cotton seeds
N. Kavitha, C. Sivaranjani and K. P. Kannan*
Bannari Amman institute of Technology, Sathyamangalam Erode District Tamil Nadu.
E-mail: kp_kanann2001@yahoo.co.in
The goal was to investigate the Cellulase production ability of fungal strains in batch culture using shake flasks. The cultures of Trichoderma viride and T.reesei were found to be the best producer of cellulases. Cellulose powder and agricultural by products such as wheat bran, sugarcane bagasse were evaluated as substrates by cultures for the production of enzymes.
Among the substrates used, natural substrates were found to be the ideal substrate providing all the nutrients for the synthesis of cellulases. Effect of concentration of cellulose powder and natural substrates, on enzyme production was studied using Folin Lowry’s method and Dinitro salicylic method. The enzyme activity was found to be maximum on 7th day for Trichoderma reesei and 8th day for Trichoderma viride. The partial purification of the cellulase enzyme on SDS PAGE revealed protein bands showing subunits of cellulase enzyme.
PS – 24
Growth responses of mycorrhizal and non-mycorrhizal bajra plants under salinity conditions
*Mahesh Borde, Mayura Dudhane and Paramjit Jite
Department of Botany, University of Pune, Pune 411007
E-mail:maheshmyco@gmail.com
Under salinity condition mycorrhizal bajra plants showed significant increase in shoot length, root length, plant fresh and dry weight. Mycorrhizal bajra plants were found to be dependent (Mycorrhizal dependency) on G. fasciculatum at second level of salinity condition. Mycorrhizal colonization was observed to be increased significantly in non saline and first level of salinity condition. After 90 days of AM inoculation mycorrhizal bajra plants showed increased photosynthetic activity upto second level of salinity. Shoot proline accumulation was higher in non mycorrhizal bajra plants and root proline accumulation was higher in mycorrhizal bajra plants. Root CAT activity was increased significantly in mycorrhizal bajra plants at second and third level of salinity, after 90 days of AM inoculation. After 60 days of AM inoculation, POD activity increased significantly in mycorrhizal bajra plants at third level of salinity (77.64%). After 90 days of AM inoculation, POD activity was found to increase significantly at all salinity levels in mycorrhizal bajra plants. Root SOD activity was found to be more in non-saline; first and second level of salinity after 60 days of AM inoculation. After 90 days, SOD activity was found to be more in second and third salinity levels in mycorrhizal bajra plants.
PS – 25
Effect of manganese concentration on growth responses of Gmelina arborea inoculated with Gigaspora margarita
Mayura Dudhane*, Mahesh Borde, and Paramjit Jite
Department of Botany, University of Pune, Pune 411 007.
*E-mail:mayuradudhane@gmail.com
Gmelina arborea Roxb. a medicinally and economically important tree species was selected for study. Mycorrhizal and nonmycorrhizal seedlings were treated with 50 mM, 100mM and 150mM concentration of Manganese. Gigaspora margarita treated Gmelina plants showed increase in shoot and root length, fresh and dry weight, greater percentage of mycorrhizal colonization, higher accumulation of proline and chlorophyll content with increasing levels of manganese. G. margarita colonization significantly increased acid and alkaline phosphatase activity, Proline content and also antioxidant enzymes like peroxidase and superoxide dismutase activity in mycorrhizal Gmelina plants as compared to non mycorrhizal plants.
PS – 26
Penicillium funiculosum rot of amla (Phyllanthus emblica L.) and its management with hot water dip treatment
Nisha Goswami* and Geeta Sumbali
Department of Botany, University of Jammu, Jammu 180 006.
*E-mail: nishagoswami23@yahoo.com
Fruits of Phyllanthus emblica L. are considered as potent gift of Mother Nature to mankind owing to their culinary usage and therapeutic potential. Approximately 15-20 % of the amla fruits are lost due to both pre- and post-harvest rot caused by Penicillium funiculosum. It is a unique pathogen requiring low pH (2.5) and low temperature (20-25oC) for its growth and the acidic fruits of amla serve as an excellent host.
Penicillium funiculosum grows aggressively, softening the fruit completely within 3-4 days, sporulating heavily on its surface and causing 100 percent loss. Healthy amla fruits get infected even by a brief contact with P. funiculosum infected fruits, suggesting thereby, the enzymatic potential of the pathogen.
In view of the extensive loss of amla fruits that occurs every year, an attempt was undertaken for the management of this disease by giving hot water dip treatment. Both pre-infection and post-infection dip treatments were given to amla fruits at 45oC, 50oC, 55oC, 60oC, 65oC, 70oC for 5 minutes. Results showed that hot water dip at 55 oC for 5 mins. gave maximum control (98.2%) without changing the texture and colour of the fruit. This method is considerably advantageous due to low cost, simplicity of application and good disease management.
PS – 27
Studies on the antagonistic effect of Bacillus sp. and Pseudomonas sp. against soil borne plant pathogenic fungi.
P. Rajapriya*, M.Pandi** and J. Muthumary**
Department of Biotechnology, Srinivasan College of Arts and Science, Perambalur 621 212.
**Centre for Advanced Studies in Botany, University of Madras, Guindy campus, Chennai 600 025.
*E-mail: pandypriya@rediffmail.com
The suppression of the growth of soil-borne plant pathogens by the rhizosphere microorganisms is termed as biocontrol. It is the reduction of the amount of inoculums or disease producing activity of pathogen accomplish by or through one or more organism. Biocontrol of plant disease with bacterial antagonist is a potential alternative of chemical control because chemical control is expensive and result in accumulation of hazardous compounds being toxic to the soil biota. However, recently some of beneficial bacterial strains applied in agriculture as biological control agents to suppress plant disease. Hence, the present study is deals with the antagonistic effect of bacillus sp. and pseudomonas sp. active against the pathogenic fungi i.e. aspergillus sp. and fusarium sp. The results will be discussed.
PS – 28
Biochemical changes in Sweet Potato infected with Charcoal Rot
V. B. Patil, A. R. Apte and S. S. Kamble*
Department of Botany, Shivaji University, Kolhapur 416 004.
*E-mail: s20sk@yahoo.co.in
Infection of sweet potato roots by M. phaseolina decreased the reducing sugars, DNA, RNA, calcium, manganese and zinc. Polyphenols were increased due to the infection by M. phaseolina isolates (sensitive and resistant). The increase was more in sweet potato roots infected by resistant isolate of M. phaseolina. Starch content was also increased in the sweet potato roots infected by sensitive and resistant isolates of M. phaseolina. There was also increase in iron content of sweet potato root due to infection by sensitive and resistant isolate of M. phaseolina.
PS – 29
Production of chitinase enzyme by Aspergillus species
Sandhya Ravishankar, C. V. Sushma, J. Sandhya, Pushpa Agrawal and Ashwani Sharma Department of Biotechnology, R.V. College of Engineering, R.V. Post, Mysore road,
Bangalore-59, Karnataka.
Fungi are used extensively to produce industrial enzymes such as lipases, amylases, cellulases, chitinases, invertases, proteases and xylanases. Among different species of fungi, Aspergillus species produces many enzymes, and the present study was aimed at production of chitinase enzyme from three different strains of Aspergillus. In vitro production of chitinases was examined under various cultural conditions. Three strains of Aspergillus were cultured in liquid synthetic medium containing different concentrations of colloidal chitin and at different incubation period. The highest specific activity of 0.257 U/mg of NAG was obtained with A.nigerS1 at 2ml concentration of colloidal chitin for 96hours incubation period. Antagonistic effect of the chitinase enzyme produced was tested by zone inhibition method for important phytopathogenic fungi (Alternaria, Fusarium, Colletotrichum and Sclerotium). The growth of these pathogens was inhibited by the action of crude extract of chitinase enzyme.
PS – 30
Ergosterol in Mucoraceous fungi
R. Rajeshwari, M. Rajeshwari, Preeti Garai, S. K. Sukrutha and J. Savitha*
Department of Microbiology & Biotechnology, Bangalore University,
Jnanabharathi Campus, Bangalore 560 056.
*E-mail: drsvtj@yahoo.co.in
Microorganisms have received broad attention as sources of novel lipids. Ergosterol [I (22E, 24R)- 24 methyl-cholesta-5,7,22, trienol] and its metabolic precursors are major lipid components of industrial and commercial interest. Ergosterol is considered as predominant sterol in fungi and yeast and is responsible for vital membrane functions such as fluidity and permeability similar to the way of cholesterol in mammalian cells. Ergosterol is the major precursor of vitamin D and is an important pharmaceutical intermediate that can be converted into cortisone and flavone hormones. Our present work involves screening of fungi belonging to the order Mucorales isolated from different natural sources for ergosterol content. The amount of ergosterol was determined using colorimetric estimation. Out of 24 isolates, the isolate 12 (Mucor sp.) showed highest ergosterol content i.e. 64mg/gm dry weight of mycelium whereas in other isolates, the ergosterol content ranged from 10mg-40mg/gm. Ergosterol in these isolates were further confirmed by HPLC analysis.
PS – 31
Exploring hidden dimension of soil fungal biodiversity: A simple technique to detect soil fungi resistant to antifungal compounds
Sonashia Velho-Pereira and Nandkumar Kamat*
Department of Botany, Goa University, Goa 403206.
*E-mail: nkamat@unigoa.ac.in
Soils are known to be ultimate and complex reservoirs of microbial diversity. The complex dimensions of bacterial and fungal diversity in tropical soils and microbial community dynamics are underexplored. Isolation techniques aimed at Actinomycetes generally employ highly selective media, powerful antibiotics and antifungal compounds to suppress undesirable bacteria and fungi. However some soil fungi may show their resistance towards these antifungal compounds. During our work to explore soil actinomycetes diversity, slides coated with Arginine Vitamin agar (AVA) incorporating a cocktail of antibiotics and antifungal compounds such as Nystatin, Cycloheximide, Terbinafin, Griseofulvin, and Flucanazole were exposed to soil environment and were retrieved at intervals of 4, 7, 15 and 28 days for detail microscopic studies of surface colonies. Alongwith actinomycetes the presence of unidentified aseptate and septate fungi was revealed indicating their resistance to combination and concentration of antifungals. Heat treatment of the soil was found to cause considerable decrease in fungal contamination probably due to elimination of heat labile fungi. Our results have led us to develop a simple procedure to sample the interesting and industrially useful strains of soil fungi resistant to common antifungal compounds. Some fungal strains are reported resistant to certain antifungals with resulting therapeutic failures as use of these antifungals inevitably selects resistant fungi, thereby pressing the urge for continuing and cyclical need of new antifungals. This technique could prove useful to detect novel antifungal resistant strains with potential to emerge as novel human pathogens. It has not escaped our notice that the probability of such finding could also help to verify whether these fungi could utilize such antifungal compounds through use of hitherto undiscovered metabolic pathways and novel enzymes leading to identification of genes responsible for antifungal resistance.
PS – 32
Biochemical investigations of fungal laccase from Pleurotus sp.
Sunil S. More* and Renuka Sudarshan
Department of Biochemistry, Center for Post Graduate studies, Jain University, 18/3,
9th Main Jayanagar, 3rd Block Bangalore 560 011.
*E-mail:sunilacr@yahoo.co.in
Laccases (benzenediol oxygen oxidoreductases, EC 1.10.3.2) are polyphenol oxidases that require O2 to oxidize phenols, polyphenols, aromatic amines and different non-phenolic substrates by one electron transfer resulting in the formation of reactive radicals.Laccasea are widely distributed in plants and fungi. In fungi they involve in morphogenesis and virulence.
In the present investigation A positive strain was isolated and characterized as non-spore forming Basidiomycetes Pleurotus sp. Laccase activity was determined using ABTS as substrate at 420 nm.Maximum laccase activity (30.8 U) was observed on 20th day in the medium with pH 6.5 laccase was purified by conventional ion exchenge chromatography. The purified laccase was a monomer showed a molecular mass of 39 ±1 kDa as estimated by SDS- PAGE with 72 fold purification with a 22 % yield. The Km and Vmax values are 250 (mM) and 0.33 (µmol/min) respectively for ABTS as substrate where as Km values for phenolic compound 2,6-DMP was 38.46 mM and Vmax 20 (µmol/min) which shows that phenolic compounds are better substrates for Pluerotus laccase than commonly used ABTS. Metal ions like CuSO4, BaCl2, MgCl2, FeCl2, ZnCl2 has no effect where as HgCl2, MnCl2 moderately decreases enzyme activity. SDS,completely inhibited enzyme where as sodium azide inhibited enzyme activity with Ki of 0.03mM and EDTA inhibited laccase to a lesser extent with a Ki of 3.2 .Urea, PCMB, DTT and Mercaptoethanol has no effect on enzyme activity.
PS – 33
Studies on Coelomycetous fungi with special reference to taxol production
S. Yogeswari*, M. Pandi, S. Kamalraj and J. Muthumary
Centre for Advanced Studies in Botany,
University of Madras, Guindy campus, Chennai 600 025.
E-mail: yogi_sagi2009@yahoo.co.in, mm_j@rediffmail.com
This paper gives an account of isolation of Coelomycetes from different places in Tamilnadu. Fifteen Coelomycetous fungi were isolated and examined. 12 were selected for screening for the production of taxol. Botryodiplodia theobromae Pat., Phomopsis sp. and Pestalotiopsis sp. were selected for taxol producing potential. Intracellular taxol was extracted from each fungus and
studied using TLC, Ultraviolet (UV) Absorption, Infrared (IR) Spectrophotometer and High Performance Liquid Chromatography (HPLC) analysis to detect Taxol. The results are presented.
POSTER SESSION 2
PS – 21
Isolation, identification and purification of endophytic fungi from medicinal plants for the enumeration of extracellular enzymes
A. Amrita, J. Swetha, P. Sindhu, and K. P. Kannan*
Department of Biotechnology, Bannari Amman Institute of technology
Satyamangalam, Erode district 638 401
*E-mail: kp_kannan2001@yahoo.co.in
Endophytes are microorganisms, which inhabit in healthy living plant tissues for all or part of their life cycle without causing apparent harmful symptoms to the host. In resent years, much research has been focused on the bioactivities of endophytic fungi. This interest is due to the possibility of these microorganisms producing pharmacologically active substances with biotechnological potential such as antitumor agents (Taxol), antifungal agents (quercine), besides producing factors of plant growth, toxins and enzymes, including some microorganisms being used as biological controllers of many diseases and plagues. In the present investigation four medicinal plants viz., Adathoda vasica, Costus ingneus, Coleus aromaticus, and Lawsonia inerims were examined for the presence of endophytic fungi. Altogether 400 segments (approximately1 cm2) of 50 segments each from leaf and stem tissues of respective plants were screened for the enumeration of the endophytic fungi using the modified surface sterilization techniques. A total of 17 different species of endophytic fungi were recorded. Among them 9 belongs to Hyphomycetes, 7 belongs to Coelomycetes and one belongs to Xylariales. From the endophytic fungi recorded, 10 dominant endophytic fungi viz., Cladosporium cladosporioides, Curvularia brachyspora, C.verruciformis, Drechslera hawaiiensis, Colletotrichum carssipes, Colletotrichum falctum, Colletotrichum gleosporioides, Lasiodiplodia theobromae, Phyllosticta Sp. and Xylariales species were tested for their ability to produce extracellular enzymes i.e., Amylase, Cellulase, Laccase, Lipase, Protease and Tyrosinase activity by the qualitative assays, majority of the entophytic fungi showed the positive results. The results will be discussed in detail.
PS – 22
Screening of endophytic fungi from the medicinal plants for the production of Chitoson
A. Amrita, P. Sindhu, J. Swetha, and K. P. Kannan*
Department of Biotechnology, Bannari Amman Institute of Technology, Satyamangalam,
Erode district 638 401.
*E-mail: kp_kannan2001@yahoo.co.in
Endophytic fungi that live inside the tissues of living plants are under-explored group of microorganisms. Recently they have received considerable attention after they were found to protect their host against insect pests, pathogens and even domestic herbivores. Medicinal plants are reported to harbour endophytes which in turn provide protection to their host from infectious agents and also provide adaptability to survive in adverse environmental conditions. It is therefore important to determine the endophytic diversity of medicinal plants. In the present study four medicinal plants were screened for the presence of Endophytic fungi and were screened for the production of chitin and Chitoson. Industrially, Chitosan is derived from the chemical deacetylation of chitin, using strong alkali, a waste product of the crustacean exoskeletons obtained after industrial processing of seafood. This process is subject to seasonal, limited supply and it pollutes the environment, due to the large amount of waste of concentrated alkaline solution. An alternative to solve these problems is the fungal production of Chitosan, whose main advantage is that it is environmentally friendly. Therefore, physicochemical properties and yields of chitosan isolated directly from a fungus may be optimised by controlling fermentation and processing parameters. The development of applications for chitosan has expanded rapidly in recent years. Chitosan, being polycationic, nontoxic, biodegradable as well as antimicrobial, has many applications especially in the agriculture, food and pharmaceutical industries. It has been used in enzyme immobilization, wastewater treatment, as food additive and anticholesterolemic, for wound healing, and in pharmaceuticals in several drug delivery systems. The results will be discussed in detail.
PS – 23
Partial purification of exracellular enzyme cellulase from Trichoderma viride and T. reesei isolated from cotton seeds
N. Kavitha, C. Sivaranjani and K. P. Kannan*
Bannari Amman institute of Technology, Sathyamangalam Erode District Tamil Nadu.
E-mail: kp_kanann2001@yahoo.co.in
The goal was to investigate the Cellulase production ability of fungal strains in batch culture using shake flasks. The cultures of Trichoderma viride and T.reesei were found to be the best producer of cellulases. Cellulose powder and agricultural by products such as wheat bran, sugarcane bagasse were evaluated as substrates by cultures for the production of enzymes.
Among the substrates used, natural substrates were found to be the ideal substrate providing all the nutrients for the synthesis of cellulases. Effect of concentration of cellulose powder and natural substrates, on enzyme production was studied using Folin Lowry’s method and Dinitro salicylic method. The enzyme activity was found to be maximum on 7th day for Trichoderma reesei and 8th day for Trichoderma viride. The partial purification of the cellulase enzyme on SDS PAGE revealed protein bands showing subunits of cellulase enzyme.
PS – 24
Growth responses of mycorrhizal and non-mycorrhizal bajra plants under salinity conditions
*Mahesh Borde, Mayura Dudhane and Paramjit Jite
Department of Botany, University of Pune, Pune 411007
E-mail:maheshmyco@gmail.com
Under salinity condition mycorrhizal bajra plants showed significant increase in shoot length, root length, plant fresh and dry weight. Mycorrhizal bajra plants were found to be dependent (Mycorrhizal dependency) on G. fasciculatum at second level of salinity condition. Mycorrhizal colonization was observed to be increased significantly in non saline and first level of salinity condition. After 90 days of AM inoculation mycorrhizal bajra plants showed increased photosynthetic activity upto second level of salinity. Shoot proline accumulation was higher in non mycorrhizal bajra plants and root proline accumulation was higher in mycorrhizal bajra plants. Root CAT activity was increased significantly in mycorrhizal bajra plants at second and third level of salinity, after 90 days of AM inoculation. After 60 days of AM inoculation, POD activity increased significantly in mycorrhizal bajra plants at third level of salinity (77.64%). After 90 days of AM inoculation, POD activity was found to increase significantly at all salinity levels in mycorrhizal bajra plants. Root SOD activity was found to be more in non-saline; first and second level of salinity after 60 days of AM inoculation. After 90 days, SOD activity was found to be more in second and third salinity levels in mycorrhizal bajra plants.
PS – 25
Effect of manganese concentration on growth responses of Gmelina arborea inoculated with Gigaspora margarita
Mayura Dudhane*, Mahesh Borde, and Paramjit Jite
Department of Botany, University of Pune, Pune 411 007.
*E-mail:mayuradudhane@gmail.com
Gmelina arborea Roxb. a medicinally and economically important tree species was selected for study. Mycorrhizal and nonmycorrhizal seedlings were treated with 50 mM, 100mM and 150mM concentration of Manganese. Gigaspora margarita treated Gmelina plants showed increase in shoot and root length, fresh and dry weight, greater percentage of mycorrhizal colonization, higher accumulation of proline and chlorophyll content with increasing levels of manganese. G. margarita colonization significantly increased acid and alkaline phosphatase activity, Proline content and also antioxidant enzymes like peroxidase and superoxide dismutase activity in mycorrhizal Gmelina plants as compared to non mycorrhizal plants.
PS – 26
Penicillium funiculosum rot of amla (Phyllanthus emblica L.) and its management with hot water dip treatment
Nisha Goswami* and Geeta Sumbali
Department of Botany, University of Jammu, Jammu 180 006.
*E-mail: nishagoswami23@yahoo.com
Fruits of Phyllanthus emblica L. are considered as potent gift of Mother Nature to mankind owing to their culinary usage and therapeutic potential. Approximately 15-20 % of the amla fruits are lost due to both pre- and post-harvest rot caused by Penicillium funiculosum. It is a unique pathogen requiring low pH (2.5) and low temperature (20-25oC) for its growth and the acidic fruits of amla serve as an excellent host.
Penicillium funiculosum grows aggressively, softening the fruit completely within 3-4 days, sporulating heavily on its surface and causing 100 percent loss. Healthy amla fruits get infected even by a brief contact with P. funiculosum infected fruits, suggesting thereby, the enzymatic potential of the pathogen.
In view of the extensive loss of amla fruits that occurs every year, an attempt was undertaken for the management of this disease by giving hot water dip treatment. Both pre-infection and post-infection dip treatments were given to amla fruits at 45oC, 50oC, 55oC, 60oC, 65oC, 70oC for 5 minutes. Results showed that hot water dip at 55 oC for 5 mins. gave maximum control (98.2%) without changing the texture and colour of the fruit. This method is considerably advantageous due to low cost, simplicity of application and good disease management.
PS – 27
Studies on the antagonistic effect of Bacillus sp. and Pseudomonas sp. against soil borne plant pathogenic fungi.
P. Rajapriya*, M.Pandi** and J. Muthumary**
Department of Biotechnology, Srinivasan College of Arts and Science, Perambalur 621 212.
**Centre for Advanced Studies in Botany, University of Madras, Guindy campus, Chennai 600 025.
*E-mail: pandypriya@rediffmail.com
The suppression of the growth of soil-borne plant pathogens by the rhizosphere microorganisms is termed as biocontrol. It is the reduction of the amount of inoculums or disease producing activity of pathogen accomplish by or through one or more organism. Biocontrol of plant disease with bacterial antagonist is a potential alternative of chemical control because chemical control is expensive and result in accumulation of hazardous compounds being toxic to the soil biota. However, recently some of beneficial bacterial strains applied in agriculture as biological control agents to suppress plant disease. Hence, the present study is deals with the antagonistic effect of bacillus sp. and pseudomonas sp. active against the pathogenic fungi i.e. aspergillus sp. and fusarium sp. The results will be discussed.
PS – 28
Biochemical changes in Sweet Potato infected with Charcoal Rot
V. B. Patil, A. R. Apte and S. S. Kamble*
Department of Botany, Shivaji University, Kolhapur 416 004.
*E-mail: s20sk@yahoo.co.in
Infection of sweet potato roots by M. phaseolina decreased the reducing sugars, DNA, RNA, calcium, manganese and zinc. Polyphenols were increased due to the infection by M. phaseolina isolates (sensitive and resistant). The increase was more in sweet potato roots infected by resistant isolate of M. phaseolina. Starch content was also increased in the sweet potato roots infected by sensitive and resistant isolates of M. phaseolina. There was also increase in iron content of sweet potato root due to infection by sensitive and resistant isolate of M. phaseolina.
PS – 29
Production of chitinase enzyme by Aspergillus species
Sandhya Ravishankar, C. V. Sushma, J. Sandhya, Pushpa Agrawal and Ashwani Sharma Department of Biotechnology, R.V. College of Engineering, R.V. Post, Mysore road,
Bangalore-59, Karnataka.
Fungi are used extensively to produce industrial enzymes such as lipases, amylases, cellulases, chitinases, invertases, proteases and xylanases. Among different species of fungi, Aspergillus species produces many enzymes, and the present study was aimed at production of chitinase enzyme from three different strains of Aspergillus. In vitro production of chitinases was examined under various cultural conditions. Three strains of Aspergillus were cultured in liquid synthetic medium containing different concentrations of colloidal chitin and at different incubation period. The highest specific activity of 0.257 U/mg of NAG was obtained with A.nigerS1 at 2ml concentration of colloidal chitin for 96hours incubation period. Antagonistic effect of the chitinase enzyme produced was tested by zone inhibition method for important phytopathogenic fungi (Alternaria, Fusarium, Colletotrichum and Sclerotium). The growth of these pathogens was inhibited by the action of crude extract of chitinase enzyme.
PS – 30
Ergosterol in Mucoraceous fungi
R. Rajeshwari, M. Rajeshwari, Preeti Garai, S. K. Sukrutha and J. Savitha*
Department of Microbiology & Biotechnology, Bangalore University,
Jnanabharathi Campus, Bangalore 560 056.
*E-mail: drsvtj@yahoo.co.in
Microorganisms have received broad attention as sources of novel lipids. Ergosterol [I (22E, 24R)- 24 methyl-cholesta-5,7,22, trienol] and its metabolic precursors are major lipid components of industrial and commercial interest. Ergosterol is considered as predominant sterol in fungi and yeast and is responsible for vital membrane functions such as fluidity and permeability similar to the way of cholesterol in mammalian cells. Ergosterol is the major precursor of vitamin D and is an important pharmaceutical intermediate that can be converted into cortisone and flavone hormones. Our present work involves screening of fungi belonging to the order Mucorales isolated from different natural sources for ergosterol content. The amount of ergosterol was determined using colorimetric estimation. Out of 24 isolates, the isolate 12 (Mucor sp.) showed highest ergosterol content i.e. 64mg/gm dry weight of mycelium whereas in other isolates, the ergosterol content ranged from 10mg-40mg/gm. Ergosterol in these isolates were further confirmed by HPLC analysis.
PS – 31
Exploring hidden dimension of soil fungal biodiversity: A simple technique to detect soil fungi resistant to antifungal compounds
Sonashia Velho-Pereira and Nandkumar Kamat*
Department of Botany, Goa University, Goa 403206.
*E-mail: nkamat@unigoa.ac.in
Soils are known to be ultimate and complex reservoirs of microbial diversity. The complex dimensions of bacterial and fungal diversity in tropical soils and microbial community dynamics are underexplored. Isolation techniques aimed at Actinomycetes generally employ highly selective media, powerful antibiotics and antifungal compounds to suppress undesirable bacteria and fungi. However some soil fungi may show their resistance towards these antifungal compounds. During our work to explore soil actinomycetes diversity, slides coated with Arginine Vitamin agar (AVA) incorporating a cocktail of antibiotics and antifungal compounds such as Nystatin, Cycloheximide, Terbinafin, Griseofulvin, and Flucanazole were exposed to soil environment and were retrieved at intervals of 4, 7, 15 and 28 days for detail microscopic studies of surface colonies. Alongwith actinomycetes the presence of unidentified aseptate and septate fungi was revealed indicating their resistance to combination and concentration of antifungals. Heat treatment of the soil was found to cause considerable decrease in fungal contamination probably due to elimination of heat labile fungi. Our results have led us to develop a simple procedure to sample the interesting and industrially useful strains of soil fungi resistant to common antifungal compounds. Some fungal strains are reported resistant to certain antifungals with resulting therapeutic failures as use of these antifungals inevitably selects resistant fungi, thereby pressing the urge for continuing and cyclical need of new antifungals. This technique could prove useful to detect novel antifungal resistant strains with potential to emerge as novel human pathogens. It has not escaped our notice that the probability of such finding could also help to verify whether these fungi could utilize such antifungal compounds through use of hitherto undiscovered metabolic pathways and novel enzymes leading to identification of genes responsible for antifungal resistance.
PS – 32
Biochemical investigations of fungal laccase from Pleurotus sp.
Sunil S. More* and Renuka Sudarshan
Department of Biochemistry, Center for Post Graduate studies, Jain University, 18/3,
9th Main Jayanagar, 3rd Block Bangalore 560 011.
*E-mail:sunilacr@yahoo.co.in
Laccases (benzenediol oxygen oxidoreductases, EC 1.10.3.2) are polyphenol oxidases that require O2 to oxidize phenols, polyphenols, aromatic amines and different non-phenolic substrates by one electron transfer resulting in the formation of reactive radicals.Laccasea are widely distributed in plants and fungi. In fungi they involve in morphogenesis and virulence.
In the present investigation A positive strain was isolated and characterized as non-spore forming Basidiomycetes Pleurotus sp. Laccase activity was determined using ABTS as substrate at 420 nm.Maximum laccase activity (30.8 U) was observed on 20th day in the medium with pH 6.5 laccase was purified by conventional ion exchenge chromatography. The purified laccase was a monomer showed a molecular mass of 39 ±1 kDa as estimated by SDS- PAGE with 72 fold purification with a 22 % yield. The Km and Vmax values are 250 (mM) and 0.33 (µmol/min) respectively for ABTS as substrate where as Km values for phenolic compound 2,6-DMP was 38.46 mM and Vmax 20 (µmol/min) which shows that phenolic compounds are better substrates for Pluerotus laccase than commonly used ABTS. Metal ions like CuSO4, BaCl2, MgCl2, FeCl2, ZnCl2 has no effect where as HgCl2, MnCl2 moderately decreases enzyme activity. SDS,completely inhibited enzyme where as sodium azide inhibited enzyme activity with Ki of 0.03mM and EDTA inhibited laccase to a lesser extent with a Ki of 3.2 .Urea, PCMB, DTT and Mercaptoethanol has no effect on enzyme activity.
PS – 33
Studies on Coelomycetous fungi with special reference to taxol production
S. Yogeswari*, M. Pandi, S. Kamalraj and J. Muthumary
Centre for Advanced Studies in Botany,
University of Madras, Guindy campus, Chennai 600 025.
E-mail: yogi_sagi2009@yahoo.co.in, mm_j@rediffmail.com
This paper gives an account of isolation of Coelomycetes from different places in Tamilnadu. Fifteen Coelomycetous fungi were isolated and examined. 12 were selected for screening for the production of taxol. Botryodiplodia theobromae Pat., Phomopsis sp. and Pestalotiopsis sp. were selected for taxol producing potential. Intracellular taxol was extracted from each fungus and
studied using TLC, Ultraviolet (UV) Absorption, Infrared (IR) Spectrophotometer and High Performance Liquid Chromatography (HPLC) analysis to detect Taxol. The results are presented.
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