Complete replacement, by biolistic plastid transformation, of the hexadecameric ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) of tobacco ( Nicotiana tabacum ) with the dimeric version from the bacterium, Rhodospirillum rubrum , resulted in fully autotrophic and reproductive tobacco plants that required high CO 2 concentrations to grow (Whitney SM, Andrews TJ ( 2001 ) Proc Natl Acad Sci USA 98: 14738-14743). Growth and photosynthesis of these plants was compared with that of nontransformed tobacco and other controls where the rbc L gene for the large subunit of tobacco Rubisco was linked to the aad A selectable-marker gene, simulating the gene arrangement of the transformants with R. rubrum Rubisco. An arrangement of the rbc L and aad A genes that gave rise to an abundant monocistronic rbc L transcript and a one-fifth as abundant bicistronic rbc L- aad A transcript had Rubisco levels and photosynthetic properties similar to those of nontransformed tobacco. Direct linkage of the rbc L and aad A genes, resulting in exclusive production of a bicistronic mRNA transcript analogous to that of the transformants with R. rubrum Rubisco, reduced transcript abundance and tobacco Rubisco content. The analogous transcript with the R. rubrum rbc M gene substituted for rbc L was not only reduced in abundance, but was also translated less efficiently. The photosynthetic rates of the transformants and controls were measured at high CO 2 concentrations, using a mass spectrometric method. The rates and their responses to atmospheric CO 2 concentration mirrored the amounts and the kinetic properties of the Rubiscos present. The contents of total nitrogen, carbohydrates, and photosynthetic metabolites of the leaves were also consistent with the content and type of Rubisco.
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