Proton and anion transport across the tonoplast vesicles in bromeliad species

Crassulacean acid metabolism (CAM) is one of the key innovations in the Neotropical family Bromeliaceae that has enabled many of its species to occupy seasonally water-limited terrestrial environments or microclimatically arid epiphytic niches. However, the relationship between CAM activity and the transport processes responsible for vacuolar organic-acid accumulation at night has not been systematically explored in this family. In the present investigation, ATP- and PPi-dependent proton transport rates were studied in tonoplast membrane vesicles isolated from leaves of six CAM and one C3 species of bromeliads. A consistent feature of these species was the high activity of the tonoplast ATP-driven H+ pump, which, when averaged across the seven species tested, showed a higher specific activity than the tonoplast PPi-driven H+ pump. For all CAM species, the rate of ATP-dependent proton transport into the tonoplast vesicles was strongly influenced by the nature of the balancing organic-acid anion, which displayed the following order of effectiveness: fumarate > malate > citrate. Measurements of leaf organic-acid content in six CAM bromeliads at dusk and dawn showed that nocturnal accumulation of malate exceeded citrate by a factor of ~2.4–20.0-fold in five of six bromeliad species used in this study, demonstrating a close correlation between the CAM rhythm and the intrinsic properties of the vacuolar membrane across which these organic acids are transported