HCO3- enrichment causes cytosolic NO3- efflux in Posidonia oceanica leaf cells

Abstract

Posidonia oceanica is a seagrass, the only group of vascular plants to colonize the marine environment. Seawater is an extreme yet stable environment, characterized by high salinity, alkaline pH and low availability of essential nutrients, such as nitrate or phosphate. In addition, in aquatic environments the supply of CO2 for the photosynthesis is limited by diffusion and therefore many aquatic plants use HCO3- as the inorganic carbon source for photosynthesis. Previous results have shown that Na+ -dependent transport systems operate on the plasma membrane of P. oceanica mesophyll leaf cells for the high-affinity NO3-, Pi or amino acids uptake. Also, a direct transport of HCO3- driven by H+ has been found in this species that provides inorganic carbon for photosynthesis and could be a significant component of a carbon concentrating mechanism in this species. Interestingly, this HCO3- direct uptake caused the efflux of chloride from the cytosol, probably through S-type anion channels, pointing that other anions could also be removed from the cytosol. This hypothesis could be relevant in the case of NO3-, since the decrease of cytosolic NO3- in response to HCO3- enrichment could limit N-assimilation. Here we analyse the effect of HCO3- increase on NO3- uptake and cytosolic homeostasis in P. oceanica. Enrichment of natural seawater with 3 mM HCO3- evokes the on-going decrease of cytosolic NO3-, from 5.7 ± 0.2 to 4.8 ± 0.7 mM after 40 min of treatment. The incubation of P. oceanica leaf pieces in 3 mM HCO3- NSW causes an initial increase of NO3- concentration in the medium. Maximum efflux (21 nmol NO3- gFM-1 min-1) occurs within the first minute of incubation. Then, external NO3- is depleted from the medium at lower net uptake rate than the value observed in non HCO3- -enriched natural seawater. These results fit the hypothesis that HCO3- enrichment causes the nitrogen loose and could impair nitrogen assimilation promoting N biomass impoverishment.