Vitamin B₁is a family of molecules, the most renowned member of which is diphosphorylated thiamine (TDP)-a coenzyme vital for the activity of key enzymes of energy metabolism. Triphosphorylated thiamine derivatives also exist within this family, specifically thiamine triphosphate (TTP) and adenosine thiamine triphosphate (ATTP). They have been investigated primarily in mammalian cells and are thought to act as metabolic messengers but have not received much attention in plants. In this study, we set out to examine for the presence of these triphosphorylated thiamine derivatives in Arabidopsis. We could find TTP in Arabidopsis under standard growth conditions, but we could not detect ATTP. Interestingly, TTP is found primarily in shoot tissue. Drivers of TTP synthesis are light intensity, the proton motive force, as well as TDP content. In plants, TTP accumulates in the organellar powerhouses, the plastids, and mitochondria. Furthermore, in contrast to other B₁vitamers, there are strong oscillations in tissue levels of TTP levels over diel periods peaking early during the light period. The elevation of TTP levels during the day appears to be coupled to a photosynthesis-driven process. We propose that TTP may signify TDP sufficiency, particularly in the organellar powerhouses, and discuss our findings in relation to its role.