Выделение и структурно-функциональная характеристика биоцидных белков из семян дыни и инжирного дерева

Аннотация

Subjects of research: melon seeds and seedlings, fig latex and dodder (Cuscuta L.). Purpose of work: Isolation, structural-functional characterization of antimicrobial and insecticidal proteins, having potential importance for the biotechnology of cotton and the other agricultural crops. Methods of research: ion exchange and hydrophobic chromatography, HPLC, partial N-tcrminal sequencing, ID and 2D PAGE, MALDI-TOF, Q-TOF MS/MS sequencing from 2D gels, protein identification using bioinformatics tools, determination of a fungicidal activity, cytotoxicity in cancer cell culture and in mice. The results obtained and their novelty: A 7597 DA protein, classified as tri-thionin, was isolated from dry seeds and seedlings of melon, while 6481 Da protein, classified as PR-protein, was isolated from the latex of fig tree. A fungicidal activity of a PR-protcin of isolated from the latex of fig tree against cotton wilt pathogens (F. oxysporium, V.dahliae) and a toxicity of melon thionin aginst plant pest larvae (Scotia segetum, Heliotis armigera, Leptinotarsa decemlineata) were determined. It is determined that a thionin fraction from the melon seeds has contained considerable amount of seed storage protein fragments, which some of them have biocidal activity. In addition to thionins and defensins plants contain other biocidal substances which could overlap with biological activity of proteins during the isolation and functional testing processes. In particular, dodder consists glycosides with cytotoxic activity in the cancer cell culture. Practical value: The methods of isolation, purification and functional testing of biocidal proteins were developed, physical and chemical characteristics of plant biocidal proteins and their properties of biological importance in agricultural biotechnology were studied. The absence of toxicity for animals demonstrate their potential significance for future development of disease- and pest - resistant transgenic plants. Degree of embed and economical effectivity: results recommended as an initial data for obtaining transgenic pathogen- and pest-resistant cotton. Field of application: agricultural biotechnology, genomics and proteomics, bioorganic chemistry, educational process.