A temporary Immersion Bioreactor System (TIB/TIS) is a fully controlled, modular biotechnical system that uses a liquid nutrient medium without agar addition. One of this system’s existing problems, however, is the emergence of vitrification (hyperhydricity), which is visually manifested in the form of plants’ morphological disorders, transparent leaf development, and swollen stems and shoots, which are deformed and become brittle. The dissertation is based on research of growing condition optimization in blueberry (Vaccinium corymbosum L.) micropropagation by a TIB/TIS system (SETISTM), with the aim of reducing harmful vitrification when applying SiO2 and ZnO nanoparticles and phloroglucinol as a plant growth regulator. The experiment was conducted at the Faculty of Agrobiotechnical Sciences Osijek during the years 2020 and 2021. The liquid nutrient medium contained the micro- and macroelements of the WPM medium (pH 5.0), with the addition of 30 g/L of sugar, 2 mg/L of zeatin, and 2 ml/L of PPM. The experiment included a control variant, four treatments using different nanoparticles (30 and 74 mg/L of nano-SiO2 and 1 and 10 mg/L of nano-ZnO, respectively), two plant growth regulator (40 and 80 mg/L of phloroglucinol) treatment, and four treatments using individual nanoparticle combinations with the plant growth regulator. At the end of the cultivation period (40 days), a significant effect of the treatment was determined on the vitrification index, leaf anatomy, morphological parameters, nutrient medium stability, mineral nutrition elements’ bioaccumulation, and blueberry plants’ physiological‐biochemical response. The vitrification index was within the range of 10 % to 40 %. The control treatment resulted in forty‐percent vitrification. By analyzing the vitrified plant material (40 %), a large deviation from a proper arrangement and structure of the leaf mesophyll cells was detected. Thus, a significantly larger stomata number was detected in the phloroglucinol treatment, as well as in the treatment of its combination with nano-SiO2 and nano-ZnO. Nano-SiO2 treatments significantly reduced the stomata number. By analyzing the morphometric and productivity parameters, nano-SiO2 treatments, and a combination with phloroglucinol, the values of a control treatment were obtained. In contrast, the nano-ZnO treatments reduced the values of morphometric parameters and multiplication, except in the case of phloroglucinol treatment with nano-ZnO, which resultn the significantly higher shoots. The decreasing concentrations of plant nutrition elements followed the element adoption dynamics within the explant plant tissue. In relation to the control treatment, the bioaccumulation of zinc (Zn) and silicon (Si) in the plant tissue is positively correlated to the values of nano-SiO2 and nano-ZnO, applied through the given treatments regardless of concentrations. The application of nano-SiO2, as well as the lower phloroglucinol levels, did not affect an increase in the total chlorophyll and carotenoid content. In contrast, the treatments of nano-ZnO, phloroglucinol, and the combination of nano-SiO2 and nano-ZnO with phloroglucinol resulted in a decrease in total chlorophyll and carotenoid content. A physiological‐biochemical response to the application of nano-SiO2 did not exert a stressful effect on the plants, and simultaneously the content of total chlorophyll and carotenoids, total phenols, and antioxidant activity and ascorbic acid did not change. Likewise, in the mentioned treatments, a low vitrification rate (20 %) and the correct morphometric and productivity proportions were established on the control level. Also, phloroglucinol, in a lower concentration of 40 mg/L, did not affect on the change of lipid peroxidation products content. The content of total chlorophyll and carotenoids, total phenols, antioxidant activity, and ascorbic acid are at the level of control treatment, with a positive effect exerted on certain morphometric parameters, productivity, and vitrification index. The research results indicate a potential of using SiO2 nanoparticles, as well as the phloroglucinol plant regulator, in suppressing the blueberry shoots’ vitrification in the TIB/TIS bioreactor under the growing conditions on a liquid nutrient medium. A protocol improvement imposes a necessity to determine the more precise concentrations and silicon forms and to determine phloroglucinol as an optional reagent model in a subsequent sub‐cultivation phase.