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Durum wheat is one of the oldest and most important edible cereal crops and its cultivation has considerable economic importance in many countries. However, adverse conditions, such as high irradiance and increasing salinity of soils, could lead to a decrease in productivity over the next few decades. Durum wheat plants under salinityare able toaccumulate glycine betaine to osmotically balance the cytosol and reduce oxidative stress, especially in young tissues. However, the synthesis of this fundamental osmolyte is inhibited by high light in T. durum even under salinity. Choline monooxygenase is the first enzyme involved in the glycine betaine biosynthetic pathway. Thus, to explain the glycine betaine inhibition, we analyzed the effect of both salinity and high light on the putative TdCMO gene expression. Thirty-eight TdCMO different transcripts were isolated in the young leaves of durum wheat grown in different stress conditions. All translated amino acid sequences, except for the TdCMO1a6 clone, showed a frame shift caused by insertions or deletions. The presence of different transcripts could depend on the presence of duplicated genes, different allelic forms, and alternative splicing events. TdCMO1a6 computational modeling of the 3D structure showed that in durum wheat, a putative CMO-like enzyme with a different Rieske type motif, is present and could be responsible for the glycine betaine synthesis.