Peroxidase activity and isoenzyme composition in Robinia pseudoacacia L. leaves under the influence of Parectopa robiniella Clemens, 1863

Іn recent times R. pseudoacacia has been suffering from populations of new pests, which cause serious economic damage to this strategic species. Therefore, studying the features of plant defense responses against harmful insects is of great importance for biodiversity conservation for preserving this species in natural and artificial plantings. The aim of the study was to analyze the peculiarities of the activity and isoenzyme composition of peroxidase in R. pseudoacacia leaves during attacks by the invading insect P. robiniella. In the green plantations of Dnipro city, as the object of research, six groups of Robinia pseudoacacia L. model trees of different ages (5 and 35–40 years of age) with the existing leaf damages by P. robiniella leaf blotch miner were identified. Differences in benzidine-peroxidase activity were found in the leaves of 5- and 40-year-old R. pseudoacacia trees. In 35‒40-year-old trees with the presence of leaf blotch mi­ners, the leaves showed a tendency to increase BPOD activity by 24.6% vs control. While in young plants, a significant increase in peroxidase activity was found by 3.8 times due to mechanical damage of the leaves by the miner compared to the control. The revealed significant increase (by 63.2%) in the activity of guaiacol-peroxidase of black locust indicates its active participation in enhancing the barrier functions of leaf cells against P. robiniella, and it indicates the promising use of the enzyme as a biomarker for pest influence. The composition of benzidine-peroxidase isoenzymes was analyzed by electrofocusing. The main pattern of changes in the expressiveness of benzidine-peroxidase due to damage of leaves by a pest was a quantitative redistribution of activity between different molecular forms of peroxidase. Several enzyme isoforms from the highly acidic pH zone showed a significant activity increase in R. pseudoacacia leaves damaged by P. robiniella; this especially applies to the peroxidase system of young black locust plants. The increase in bensidine- and guajacol-peroxidase activity under the influence of leaf miner may be explained by a change in the gene expression of individual isoforms of peroxidases, which is confirmed by the results of studying the relative share of peroxidase molecular forms with isoelectrofocusing. Calculation of the internal diversity index (KD) of IEF benzidine-peroxidase spectra showed that the studied peroxidase spectra of young and medieval trees of control and experimental samples of R. pseudoacacia leaves differ significantly in the level of complexity. Under the influence of the invader the level of complexity increased. The study of the reaction of black locust and the mechanisms of its protection against mechanical damage by phytophages with the involvement of a larger range of enzymes in further research can serve as a theoretical basis for species selection and management to restore it under biotic stress.