Tocopherol metabolism, oxidation and recycling under high light stress in Arabidopsis.

CONCLUSIONS

Tocopherols are synthesized and accumulated by all plants and many cyanobacteria. The quenching and scavenging of reactive oxygen species and lipid peroxy radicals by tocopherols can result in the formation of various tocopherol oxidation compounds. A targeted GC/MS profiling method was developed to quantify all tocopherols and pathway intermediates, and 23 potential alpha- and gamma-tocopherol oxidation products. This method was used to study the response of wild-type Arabidopsis (Col) and the tocopherol biosynthetic mutants vte1, vte2 and vte4 during 12 h low- and high-light treatments (LL and HL, 90 and 1500 mumol photon m(-2) sec(-1), respectively) and a subsequent 12 h dark recovery period. All tocopherols and pathway intermediates exhibited HL-dependent increases except 2,3-dimethyl-6-phytyl-1,4-benzoquinone (DMPBQ) in vte1 and beta-tocopherol in Col. Profiling of potential tocopherol oxidation products during HL treatment indicated the presence of only alpha-tocopherolquinol (alpha-TQH(2)) in Col and only gamma-tocopherolquinol (gamma-TQH(2)) in vte4, both of which accumulated to similar levels and with similar kinetics the two genotypes. However, during dark recovery, the level of alpha-TQH(2) in Col decreased several times faster than that of gamma-TQH(2) in vte4, suggesting the presence of biochemical processes with higher specificity for alpha-TQH(2). (14)C-labeled alpha-tocopherolquinone (alpha-TQ) applied to isolated Col chloroplasts was converted to (14)C-alpha-tocopherol, demonstrating the existence of a plastid-based system for recycling oxidized alpha-tocopherol. The accumulation of (14)C-trimethylphytylbenzoquinone (TMPBQ) by isolated vte1 plastids treated with (14)C-labeled alpha-TQ is consistent with the tocopherolquinone-recycling pathway utilizing a yet to be identified plastid-localized dehydratase that converts tocopherolquinone to TMPBQ.