Relocation of the plastid rbcL gene to the nucleus yields functional ribulose-1,5-bisphosphate carboxylase in tobacco chloroplasts.

Abstract

The conserved plastid localization of rbcL suggests that biosynthesis of the large subunit of ribulose-1,5-bisphosphate carboxylase [Rubisco; 3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39] in chloroplasts is required to obtain functional enzyme. To examine the validity of this hypothesis, we relocated the plastid rbcL gene to the nucleus. First, we deleted the rbcL gene from the tobacco plastid genome by targeted insertion of a selectable aadA gene encoding spectinomycin resistance. The rbcL coding region was then inserted into an expression cassette and introduced into the nuclear genome of these plants by Agrobacterium-mediated transformation. We report that the nuclear rbcL functionally complements the defective plastids when the Rubisco large subunit is targeted to chloroplasts by a transit peptide. Therefore, the evolutionary process that relocates functional plastid genes to the nucleus has not yet occurred in the case of the rbcL gene. Targeted deletion of plastid genes, combined with their allotopic expression, will provide opportunities for studying the function of plastid enzyme complexes.