Properties , Physiological Role and Possible Use in Biotechnology of Proteinase Inhibitor from Buckwheat Seeds

Isolation and detailed investigation of protein proteinase inhibitor BWI-1a from buckwheat seeds has been carried out. The protein obtained was homogeneous and its physico-chemical, kinetic and biological properties were characterized. The molecular mass of BWI1a inhibitor was 7.7 kDa and it consisted of 69 amino acid residues. Interaction of the studied inhibitor with a number of proteinases secreted by pathogenic filamentous fungi has been studied. The results obtained made it possible to propose that the inhibitor participated in the defense system of buckwheat seeds. Further complete amino acid sequence of BWI-1a inhibitor was established, the gene of the inhibitor was obtained, and genetic engineered constructions were created for transformation of some plants. Finally, transgenic potato and tobacco plants, containing BWI-1a inhibitor gene, were obtained, which demonstrated increased resistance to 2 bacterial phytopathogens Pseudomonas syringae pv. tomatoe and Clavibacter michiganensis sbsp. michiganensis. Biotest consisting in damaging of tobacco plants with white wings butterfly Trialeurodes vaporariorum in the greenhouse also demonstrated the existence of defense effect in transgenic tobacco plants. Thus, obtained results confirmed the proposal on the defense role of proteinase inhibitor BWI-1a and point to the possibility of its use for production of transgenic plants with increased resistance to phytopathogens and insect pests. _____________________________________________________________________________________________________________

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