Localization and activity of lipoxygenase in the ovule of Larix kaempferi (Lamb.) Carr. during female gametophyte maturation.

ABSTRACT

KEY MESSAGE Lipoxygenase activity and localization vary throughout the development of Larix kaempferi ovules, with the highest enzyme activity observed in ovules at the cellular stage and the most intense immunogold reaction noted at the mature archegonium stage of gametophyte development. Lipoxygenases are a family of oxidoreductases with a significant role in biological systems, widespread in living organisms e.g. mammals, fish, corals, plants, mosses, algae, fungi, yeasts, and bacteria. Lipoxygenase activity in plants leads to the formation of phytooxylipins, i.e. signaling molecules, which play a crucial role in many significant physiological processes such as male and female gametophyte maturation, germination and seedling growth, pathogen resistance, abiotic stress response, fruit ripening, and senescence. The activity and localization of lipoxygenase change during plant growth and development. The localization of lipoxygenase in a developing ovule of Larix kaempferi was analyzed using the immunogold labeling method, and the activity was determined spectrophotometrically with linolenic acid as a substrate. Among the investigated stages, the immunogold reaction was the most intense at the mature archegonium stage in the ovule. Lipoxygenase was found in all parts of the L. kaempferi ovule. The largest number of immunogold particles was detected in the integument cells of all the analyzed stages of ovule development. Only one isoform of lipoxygenase with an optimum at pH 8 was active in the ovules during female gametophyte maturation. The highest enzyme activity was determined at the cellular stage, whereas the mature archegonium stage was characterized by its lowest level, which means that LOX activity in developing ovules of the Japanese larch is not correlated with the number of antibody-labeled molecules of the enzyme.