Structural and functional features of the leaves of Ranunculus trichophyllus Chaix., a freshwater submerged macrophophyte
AZA, 5-Acetamido-1,3,4-thiadiazole-2-sulphonamide 
CA, carbonic anhydrase 
DIC, dissolved inorganic carbon 
Hepes, A-(2-hydroxyethyl)-1 piperazine-ethane sulfonic acid 
IC, inorganic carbon 
PAR, photosynthetic active radiation 
PATAg, periodic acid-thiosemicarbazide-silver proteinate 
Tris, tris (hydroxymethyl)-aminomethane 
 
The structural and physiological strategies developed by the leaves of the freshwater macrophyte Ranunculus trichophyllus to adapt to submersed life were studied. Photosynthesis is carried out mainly by the epidermis cells of the numerous segments into which the leaf is finely dissected. In these cells, containing most of the chloroplasts, a peculiar organization of the wall has been identified by cytochemical tests. A thin compact outer region covers the cell surface and splits up forming large lacunae between adjacent cells. Below it, a thick and loose inner region rich in hydrophilic pectic acids occurs, which grows in along the cell sides giving rise to wide transfer areas. In this latter cell wall region, in which the cell/environment contact and exchanges are amplified, the systems for inorganic carbon supply to photosynthetic cells operate. The leaves of R. trichophyllus can rely on environmental CO2 and HCO3– as sources of inorganic carbon for photosynthesis. A mechanism for bicarbonate utilization seems to involve its conversion to CO2 by an apoplastic carbonic anhydrase, whose activity gains importance as the availability of environmental CO2 decreases. Interestingly, it has been demonstrated that in this species CO2 can also be obtained from HCO3– by a photodependent increase in plasmamembrane H+-ATPase activity in the transfer areas of the epidermis cells. This is the first time that such a mechanism has been noted in a nonpolar leaf of a submerged macrophyte.