Kovalyova V. Structural and functional characteristics of Scots pine defensin 1 (Pinus sylvestris L.): isolation, identification, cloning of the gene and biological activity.

Object - molecular mechanisms, which a Scots pine resistance into phytopathogenic fungus are providing. Aim of the study - to isolate a Scots pine defensin 1 (PsDef1) and to study its structural and functional features and also to clone cDNA of defensin 1, to obtain its recombinant product and antibody against defensin. Methods: biochemical, gene engineering, microbiological, statistical. A new defensin-like protein have been identified in Scots pine seedling tissues. PsDef1 was purified and was shown to possess high antifungal activity toward phytopathogenic fungi. cDNA library of 7-day old pine root seedlings was created. Clones which encoded two pine defensins, named by us PsDef1 and PsDef2, was isolated from this library. Nucleotide sequences of these proteins were deposited in Genbank. Structural and functional similarity of PsDef1 with plant defensins of the first (morphogenic) group was shown. Functionally active recombinant Psdef1 and polyclonal antibodies were generated and used in the research of PsDef1expression in pine seedling tissues during seed germination and under phytopathogenic fungus Heterobasidion annosum infection. Area of use - biochemistry, phytopathology, plant physiology.