Interaction of elevated CO2 and form of nitrogen nutrition alters leaf abaxial and adaxial epidermal and stomatal anatomy of wheat seedlings

Plant's stomatal physiology and anatomical features are highly plastic and are influenced by diverse environmental signals including the concentration of atmospheric CO2 and nutrient availability. Recent reports suggest that the form of nitrogen (N) is a determinant of plant growth and nutrient nitrogen use efficiency (NUE) under elevated CO2 (EC). Previously, we found that high nitrate availability resulted in early senescence, enhanced reactive oxygen species (ROS), and reactive nitrogen species (RNS) production and also that mixed nutrition of nitrate and ammonium ions were beneficial than sole nitrate nutrition in wheat. In this study, the interactive effects of different N forms (nitrate, ammonium, mixed nutrition of nitrate, and ammonium) and EC on epidermal and stomatal morphology were analyzed. Wheat seedlings were grown at two different CO2 levels and supplied with media devoid of N (N0) or with nitrate-N (NN), mixed nutrition of ammonium and nitrate (MN), or only ammonium-N (AN). The stoma length increased significantly in nitrate nutrition with a consistent reduction in stoma width. Guard cell length was higher in EC treatment as compared to AC. The guard cell width was maximum in AN-grown plants at EC. Epidermal cell density and stomatal density were lower at EC. Nitrate nutrition increased the stomatal area at EC while the reverse was true for MN and AN. Wheat plants fertilized with AN showed a higher accumulation of superoxide radical (SOR) at EC, while in NN treatment, the accumulation of hydrogen peroxide (H2O2) was higher at EC. Reactive oxygen species, particularly H2O2, can trigger mitogen-activated protein kinase (MAPK) mediated signaling and its crosstalk with abscisic acid (ABA) signaling to regulate stomatal anatomy in nitrate-fed plants. The SOR accumulation in ammonium- and ammonium nitrate-fed plants and H2O2 in NN-fed plants might finely regulate the sensitivity of stomata to alter water/nutrient use efficiency and productivity under EC. The data reveals that the variation in anatomical attributes viz. cell length, number of cells, etc. affected the leaf growth responses to EC and forms of N nutrition. These attributes are fine targets for effective manipulation of growth responses to EC.