Purine metabolism during white spruce somatic embryo development: salvage of adenine, adenosine, and inosine

Abstract

Contribution of the adenine, adenosine and inosine salvage to the purine nucleotide and nucleic acid biosynthesis during white spruce (Picea glauca) somatic embryo maturation was estimated by in situ assays using [8-(14)C]adenine, [8-(14)C]adenosine and [8-(14)C]inosine. The salvage of adenine and adenosine was high during the initial stages of embryo maturation, characterized by rapid cell proliferation, but it declined upon further embryo development. Inosine salvage activity was always much lower than that observed for adenine and adenosine. Consistent with these results, activities of adenine phosphoribosyltransferase (APRT) and adenosine kinase (AK) measured in the embryo extracts in vitro were much higher than the activity of inosine kinase (IK) during all stages of embryo development. Utilization of adenosine and inosine for nucleotide and nucleic acid synthesis was found to be regulated by the enzymes AK and IK, as the pattern of their activities was very similar to the activity of adenosine and inosine salvage, estimated with exogenously supplied precursors. However, little correlation between salvage of adenine and activity of APRT was found throughout somatic embryo maturation. As no adenosine nucleosidase activity was found in white spruce embryos, adenosine, but not adenine, seems to be the major end product of adenylate catabolism and becomes the predominant substrate for purine salvage in vivo. Thus, adenosine salvage appeared to have the most important role in white spruce embryos. Studies on the metabolic fate of [8-(14)C]adenine and [8-(14)C]adenosine suggest that turnover of adenine nucleotides is rapid, as some of them are utilized for nucleic acid synthesis. In contrast, most of [8-(14)C]inosine taken up by the embryos seems to be directly catabolized by the conventional purine catabolic pathway via ureides in all stages of embryo maturation.