INFLUENCE OF STERILANT SUBSTANCES ON THE DECONTAMINATION OF PINUS SYLVESTRIS L. EXPLANTS IN CULTURE IN VITRO

Abstract

An analysis and generalization of a number of literary data, both domestic and foreign scientists, concerning achievements in the field of microclonal propagation in in vitro culture of Pinus sylvestris L. plants, in particular the process of decontamination of initial explants, was carried out. A study was carried out on the influence of the concentration of chemical sterilant agents, as well as the duration of treatment (exposure) with them on the sterilization of initial explants of the studied species. The used methodology for conducting experimental studies on obtaining an aseptic culture of Pinus sylvestris L. explants for their further propagation in vitro was described in detail. A stepwise scheme of decontamination of plant material was tested, which took place in two main stages: primary and modified sterilization. The first stage consisted of treating the explants as follows: running water with an appropriate detergent («Tween 80»), hydrogen peroxide (H2O2) (1; 2; 3% and 3; 5; 10 minutes), ethyl alcohol (C2H5OH) (50; 70; 96% and 5; 10; 15 seconds) in the indicated concentrations and exposures. Modification of the primary scheme of decontamination of explants was performed using the following basic chemicals: NaClO (10; 20; 30% and 3; 5; 7 minutes), AgNO3 (0.1; 0.2; 0.3 % and 5; 10; 15 minutes) and HgCl2 (1; 2; 3% and 5; 10; 15 minutes). To prevent the development of internal infections, 0.1% antibiotic «Imanin» was used. A study was conducted of the specific individual effect of the main chemical sterilants on the sterilization efficiency and viability of Pinus sylvestris L. explants. The optimal stepwise decontamination scheme for explants of the studied species was experimentally established, which provided sterilization efficiency of 94.33±1.41% and explant viability of 91.7±1.34% and consisted of alternate treatment of plant material as follows: С2Н5ОН (96%; 10 seconds) + Н2О with detergent (6 h) + Н2О2 (3%; 10 minutes) + С2Н5ОН (70 %; 10 s) + AgNO3 (0.2%; 5 minutes) + NaClO (30%; 5 minutes).