Application of a System for Targeting Fe-Dependent Superoxide Dismutase and Choline Oxidase Enzymes to Chloroplast as a Strategy for Effective Plant Resistance to Abiotic Stresses

Abstract

The plastids involved in photosynthesis and other essential metabolic processes are the cellular compartments most susceptible to abiotic stress factors with adverse effects in plants. In order to protect the chloroplast compartment, the plants of the Belyi Naliv tomato cultivar and the Darenka potato cultivar were transformed with genes for Fe-dependent superoxide dismutase and choline oxidase with the use of the method for plant genetic engineering. These target genes were equipped with signal sequences directing the proteins into the plastid compartment. The experimental results have demonstrated the optimization of the agrobacterium-mediated transformation method for the analyzed tomato and potato cultivars. The naturally growing leaves and stems of the mature plants proved to be the most effective explants to obtain the regenerants. The highest regeneration frequency in the explants was reached with the use of a medium containing 5 mg L^–1 BAP, 0.1 mg L^–1 NAA, and 1 mg L^–1 zeatin. Augmentin and timentin were equally efficient among the antibiotics tested for the inhibition of the agrobacterium; the concentration of 200 mg L^–1 in the culture medium was sufficient to eliminate the agrobacterium. In addition, they had no inhibitory effects on the shoot regeneration process. However, the cefotaxime concentration required to inhibit the agrobacterium should be 500 mg L^–1 and higher. Moreover, this antibiotic at the concentration of 100 mg L^–1 was proven to inhibit the regenerant growth.