Formate-14C was fed to detached primary leaves of wheat and the distribution of activity among various fractions was examined after 2 to 26 h of metabolism in the light or 4 h of metabolism in the dark.All samples contained activity in free neutral, acidic, and basic compounds. The radioactive metabolites in the basic fraction were examined in detail. Initially, in the light, glutamate, aspartate, serine, and an unknown, chromatographically similar to, but not identical with histidine, contained most of the activity. Activity in betaine increased with time until, at 26 h, it contained nearly half of the activity in this fraction. Following dark metabolism, most of the activity resided in glutamate and serine. Glycine was not radioactive, and most of the activity in serine resided in carbon 3, indicating that formate served as a precursor of C1-units that were used for serine synthesis. Activity was also detected in a number of other amino acids and choline, both in the light and dark.A comparison of formate-14C metabolism in the dark in stem rust resistant and susceptible wheat leaves revealed that betaine, lipids, nucleic acid bound adenine and guanine, and a protein-containing residue from rust-infected susceptible leaves contained much more activity than the corresponding components from healthy susceptible or from healthy or rust-infected resistant leaves.Activity from glycine-2-3H, methionine-14CH3, serine-3-14C, ethanolamine-1,2-14C, and choline-14CH3 was incorporated into betaine in the dark, but was not detected in sarcosine or dimethylglycine. These results support the view that betaine was synthesized from glycine via serine, ethanolamine, and choline with methionine as the methyl donor, and not by direct N-methylation of glycine.Betaine-14CH3 was translocated but not metabolized in healthy or rust-infected primary leaves of wheat, or in aerial portions of adult wheat plants. In these wheat tissues, betaine would thus appear to be a metabolic end product.