Antifungal properties of haem peroxidase from Acorus calamus.
OBJECTIVE
Plants have evolved a number of inducible defence mechanisms against pathogen attack, including synthesis of pathogenesis-related proteins. The aim of the study was to purify and characterize antifungal protein from leaves of Acorus calamus.


METHODS
Leaf proteins from A. calamus were fractionated by cation exchange chromatography and gel filtration and the fraction inhibiting the hyphal extension of phytopathogens was characterized. The temperature stability and pH optima of the protein were determined and its presence was localized in the leaf tissues.


RESULTS
The purified protein was identified as a class III haem peroxidase with a molecular weight of approx. 32 kDa and pI of 7.93. The temperature stability of the enzyme was observed from 5 degrees C to 60 degrees C with a temperature optimum of 36 degrees C. Maximum enzyme activity was registered at pH 5.5. The pH and temperature optima were corroborated with the antifungal activity of the enzyme. The enzyme was localized in the leaf epidermal cells and lumen tissues of xylem, characteristic of class III peroxidases. The toxic nature of the enzyme which inhibited hyphal growth was demonstrated against phytopathogens such as Macrophomina phaseolina, Fusarium moniliforme and Trichosporium vesiculosum. Microscopic observations revealed distortion in the hyphal structure with stunted growth, increased volume and extensive hyphal branching.


CONCLUSIONS
This study indicates that peroxidases may have a role to play in host defence by inhibiting the hyphal extension of invading pathogens.
Ghosh
Modhumita
M
Division of Plant Biotechnology, Institute of Forest Genetics and Tree Breeding Forest Campus, R.S. Puram, Coimbatore--641 002, India.
D029023:Acorus / Q000201:enzymology*; D000595:Amino Acid Sequence; D000908:Antibodies, Fungal / Q000187:drug effects; D000935:Antifungal Agents / Q000378:metabolism / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010544:Peroxidases / Q000378:metabolism / Q000494:pharmacology*; D010935:Plant Diseases / Q000382:microbiology; D018515:Plant Leaves / Q000201:enzymology
D000908:Antibodies, Fungal; D000935:Antifungal Agents; D010544:Peroxidases