Abstract
Objective: This research aimed to verify the use of aqueous ozone as an alternative sterilizer for in vitro sterilization procedure.
 Methods: Three species of Solanaceae were selected based on their trichomes. They are thorn apple (Datura metel L.), eggplant (Solanum melongena), and tomato (Lycopersicon pimpinellifolium). The treatment used four levels of concentrations; K0 (NaOCl – control), K1 (AO 0.1 ppm), and K2 (AO 0.5 ppm), K3 (1 ppm) and it employed three types of leaves, namely, D1 (thorn apple), D2 (eggplant), and D3 (tomato). The data were analyzed by analysis of variance univariate two factors and followed by Duncan’s multiple comparisons test.
 Results: This assay showed that the effect of ozone was almost immediately against microorganisms, and the optimum ozone concentration depends on the types of the leaf. Both levels of ozone concentration and types of leaves had a significantly different effect (p<0.05), and there is no interaction between aqueous ozone concentration and types of leaves (p>0.05). The level of sterility in K0 is 77, 78%, K1 is 22, 22%, K2 is 44, 44%, and K3 is 100%. Ozone 1 ppm had the highest result, but there is no viable explant because they got damage in their cells.
 Conclusions: Aqueous ozone showed good potential to inhibit the growth of microorganisms for the in vitro sterilization procedure of Solanaceae.
Highlights
Plant tissue culture or known as in vitro culture is an effort to multiply plants by isolating plant parts and grow them in aseptic conditions [1,2,3], these parts can multiply and become selfpropagating complete plants [4]
The results of the analysis of variance (ANOVA) test at a 95% confidence level indicate that there is no interaction between the leaf types with ozone
It means the explants derived from eggplant leaves give the highest level of sterility when compared with thorn apple and tomato leaves
Summary
Plant tissue culture or known as in vitro culture is an effort to multiply plants by isolating plant parts and grow them in aseptic conditions [1,2,3], these parts can multiply and become selfpropagating complete plants [4]. Contamination with microorganisms has considered being the single most important reason for losses during in vitro culture of plants [8]. Such microorganisms include viruses, fungi, bacteria, and yeast [9]. Fungi, bacteria, and yeast [9] The presence of these microorganisms usually resulting in increased mortality and invariable growth, necrosis, reduced proliferation, and reduced rooting. Efforts to maintain sterile conditions are necessary for a whole series of in vitro culture activities, so sterilization must be done to prevent and reduce contamination [10]
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