A search for antiplasmodial metabolites among fungal endophytes of terrestrial and marine plants of southern India

Eighty four different fungal endophytes isolated from sea grasses (5), marine algae (36) and leaves or barks of forest trees (43) were grown in vitro and the secondary metabolites secreted by them were harvested by immobilizing them on XAD beads. These metabolites were eluted with methanol and screened using SYBR Green I assay for their antiplasmodial activity against blood stage Plasmodium falciparum in human red blood cell culture. Our results revealed that fungal endophytes belonging to diverse genera elaborate antiplasmodial metabolites. A Fusarium sp. (580, IC50: 1.94 μg ml−1) endophytic in a marine alga and a Nigrospora sp. (151, IC50: 2.88 μg ml−1) endophytic in a tree species were subjected to antiplasmodial activity-guided reversed phase high performance liquid chromatography separation. Purification led to potentiation as reflected in IC50 values of 0.12 μg ml-1 and 0.15 μg ml−1 for two of the fractions obtained from 580. Our study adds further credence to the notion that fungal endophytes are a potential storehouse for a variety of novel secondary metabolites vested with different bioactivities including some that can stall the growth of the malaria parasite.

[1]  J. Keasling,et al.  High-level semi-synthetic production of the potent antimalarial artemisinin , 2013, Nature.

[2]  S. Jennewein,et al.  Getting to the bottom of Taxol biosynthesis by fungi , 2013, Fungal Diversity.

[3]  John A. Johnson,et al.  Antimicrobials from the Marine Algal Endophyte Penicillium sp , 2013, Natural product communications.

[4]  S. Nayak.  Do endophytic fungi possess pathway genes for plant secondary metabolites? , 2013 .

[5]  T. S. Murali.  Fungal communities of symptomless bark of tropical trees , 2013 .

[6]  P. Newton,et al.  A Major Genome Region Underlying Artemisinin Resistance in Malaria , 2012, Science.

[7]  John L. Spouge,et al.  Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi , 2012, Proceedings of the National Academy of Sciences.

[8]  Jay D. Keasling,et al.  Production of amorphadiene in yeast, and its conversion to dihydroartemisinic acid, precursor to the antimalarial agent artemisinin , 2012, Proceedings of the National Academy of Sciences.

[9]  M. Nei,et al.  MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. , 2011, Molecular biology and evolution.

[10]  S. Stephenson,et al.  From morphology to molecular biology: can we use sequence data to identify fungal endophytes? , 2011, Fungal Diversity.

[11]  J. Clardy,et al.  New naphthoquinones and a new δ-lactone produced by endophytic fungi from Costa Rica. , 2011, Tetrahedron letters.

[12]  P. Proksch,et al.  Fungal endophytes: unique plant inhabitants with great promises , 2011, Applied Microbiology and Biotechnology.

[13]  M. Tejesvi,et al.  Potential of Tree Endophytes as Sources for New Drug Compounds , 2011, Endophytes of Forest Trees.

[14]  R. Sukumar,et al.  Endophytic fungal communities in woody perennials of three tropical forest types of the Western Ghats, southern India , 2011, Biodiversity and Conservation.

[15]  T. Anke,et al.  Antimicrobial Compounds from Tree Endophytes , 2011 .

[16]  A. Pirttilä,et al.  Endophytes of forest trees : biology and applications , 2011 .

[17]  S. Meshnick,et al.  Antimalarial drug resistance of Plasmodium falciparum in India: changes over time and space. , 2011, The Lancet. Infectious diseases.

[18]  M. Doble,et al.  Internal mycobiota of marine macroalgae from the Tamilnadu coast: distribution, diversity and biotechnological potential , 2010 .

[19]  Wolfgang Maier,et al.  Current state and perspectives of fungal DNA barcoding and rapid identification procedures , 2010, Applied Microbiology and Biotechnology.

[20]  P. Proksch,et al.  Methods for isolation of marine-derived endophytic fungi and their bioactive secondary products , 2010, Nature Protocols.

[21]  Tim Anderson,et al.  Mapping the Spread of Malaria Drug Resistance , 2009, PLoS medicine.

[22]  F. Sasse,et al.  Fungal endophytes and bioprospecting , 2009 .

[23]  D. Weber.  Endophytic Fungi, Occurrence and Metabolites , 2009 .

[24]  U. Pinruan,et al.  Marine endophyte sources of new chemical natural products: a review , 2008 .

[25]  N. White,et al.  Qinghaosu (Artemisinin): The Price of Success , 2008, Science.

[26]  C. Raghukumar.  Marine fungal biotechnology: An ecological perspective , 2008 .

[27]  K. Siems,et al.  Screening strategies for obtaining novel, biologically active, fungal secondary metabolites from marine habitats , 2008 .

[28]  W. Nierman,et al.  The effect of elevated temperature on gene transcription and aflatoxin biosynthesis , 2007, Mycologia.

[29]  W. Nierman,et al.  The effect of elevated temperature on gene transcription and aflatoxin biosynthesis. , 2007 .

[30]  R. Henrik Nilsson,et al.  Taxonomic Reliability of DNA Sequences in Public Sequence Databases: A Fungal Perspective , 2006, PloS one.

[31]  A. Wright,et al.  Potential antimalarial lead structures from fungi of marine origin. , 2005, Planta medica.

[32]  Weltgesundheitsorganisation.  World malaria report , 2005 .

[33]  A Ganesan,et al.  Natural products and combinatorial chemistry: back to the future. , 2004, Current opinion in chemical biology.

[34]  P. Wilairat,et al.  Simple and Inexpensive Fluorescence-Based Technique for High-Throughput Antimalarial Drug Screening , 2004, Antimicrobial Agents and Chemotherapy.

[35]  P. Roepe,et al.  Novel, Rapid, and Inexpensive Cell-Based Quantification of Antimalarial Drug Efficacy , 2004, Antimicrobial Agents and Chemotherapy.

[36]  C. Ireland,et al.  Marine-derived fungi: a chemically and biologically diverse group of microorganisms. , 2004, Natural product reports.

[37]  P. Kongsaeree,et al.  Antimalarial dihydroisocoumarins produced by Geotrichum sp., an endophytic fungus of Crassocephalum crepidioides. , 2003, Journal of natural products.

[38]  Axel Zeeck,et al.  Big Effects from Small Changes: Possible Ways to Explore Nature's Chemical Diversity , 2002, Chembiochem : a European journal of chemical biology.

[39]  P. Kittakoop,et al.  Antimalarial halorosellinic acid from the marine fungus Halorosellinia oceanica. , 2001, Bioorganic & medicinal chemistry letters.

[40]  B. Schulz,et al.  The endophyte-host interaction: a balanced antagonism? , 1999 .

[41]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[42]  R. Pereira,et al.  Polyphenols, Terpenes and Sterols in Brazilian Dictyotales and Fucales (Phaeophyta) , 1994 .

[43]  T. Bruns,et al.  ITS primers with enhanced specificity for basidiomycetes ‐ application to the identification of mycorrhizae and rusts , 1993, Molecular ecology.

[44]  J. Hamilton-miller,et al.  Comparative study of cephradine and amoxicillin-clavulanate in the treatment of recurrent urinary tract infections , 1990, Antimicrobial Agents and Chemotherapy.

[45]  Thomas J. White,et al.  PCR protocols: a guide to methods and applications. , 1990 .

[46]  T. White.  Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics , 1990 .

[47]  N. Saitou,et al.  The neighbor-joining method: a new method for reconstructing phylogenetic trees. , 1987, Molecular biology and evolution.

[48]  J. Felsenstein.  CONFIDENCE LIMITS ON PHYLOGENIES: AN APPROACH USING THE BOOTSTRAP , 1985, Evolution; international journal of organic evolution.

[49]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[50]  C. Lambros,et al.  Synchronization of Plasmodium falciparum erythrocytic stages in culture. , 1979, The Journal of parasitology.

[51]  W. Trager,et al.  Human malaria parasites in continuous culture. , 1976, Science.

[52]  W. J. Humphreys.  SECTION B, PHYSICS. , 1916, Science.