Метод стерилізації агару та живильних середовищ для біотехнологічних досліджень в культурі in vitro

Abstract

Purpose. To develop a method of sterilization of agar and nutrient media for the propagation and cultivation of in vitro plant material using a microwave oven. Methods. Laboratory, biotechnological, physiological, analytical. Results. The research has identified the possibility of using a microwave oven to sterilize nutrient media and agar. The influence of sterilization conditions in a microwave oven (terms, mode and power of microwave radiation) on the quality of nutrient media — sterility, density, infection was analyzed. The optimal conditions of sterilization in a microwave oven was established: in a continuous mode 2–3 minutes after boiling of a liquid (temperature about 100 °C) at 750–900 W; in a throbbing 30 seconds at 750–900 W + 1–2 minutes at 500 W + 30 seconds at 750–900 W or 30 seconds + 30 seconds + 30 seconds at 900 W, which ensure the production of sterile qualitative media in a short time. While in autoclaves, agar and nutrient media was sterilized at temperatures above 100 °C (130–140 °C) for 120 minutes, which does not ensure the preservation of the biological value of sugars, vitamins, growth regulators, amino acids. According to the developed method of sterilization, more than 30 different compositional nutrient media were produced for microclonal propagation of miscanthus, obtaining morphogenic callus and embryos of sugar beet by the method of androgenesis. in vitro. Conclusions. The method of sterilization of nutrient media for the propagation and cultivation of in vitro plant material by thermal treatment using a microwave oven allows the preparation of nutrient media in much shorter terms (2–3 minutes) than when using an autoclave (120 minutes) and provides sterility and the preservation of the biological value of organic and mineral substances and the costeffectiveness of the process.