Choride as a beneficial macronutrient in plants: biological functions and regulation

1. Background and Objectives: In the agronomic context, chloride (Cl-) has been generally considered a toxic anion rather than a plant nutrient (Brumós et al, 2010). However we have recently shown that, in addition to an essential micronutrient, Cl- is a beneficial macronutrient (Franco-Navarro et al, 2016). Under non-saline conditions (1-5 mM), Cl- specifically stimulates higher leaf cell size and leads to a moderate increase of plant fresh and dry biomass mainly due to higher shoot expansion. Chloride plays specific roles in regulating leaf osmotic potential and turgor, allowing plants to improve leaf water balance parameters. In addition, Cl- regulates water relations at the whole plant level through reduction of plant transpiration. This is a consequence of a lower stomatal conductance, which results in lower water loss and greater photosynthetic and integrated water-use efficiency (Franco-Navarro et al, 2016). 2. Material and Methods: Tobacco plants were predominantly used for physiological measurements; Arabidopsis thaliana plants were used for the molecular characterization of genes involved in Cl- transport. Physiological methods used in this work include quantification of: nutrients content; plant biomass; anatomic parameters (leaf area, leaf cell size and density, chloroplast size and density and stomatal opening); water parameters (water content, relative water content, succulence, water potential, osmotic potential, turgor and water use efficiency); photosynthetic parameters (photosynthetic rate, stomatal conductance, intrinsic WUE and mesophyll conductance to CO2). Molecular methods used in this work include: tissue and cell-specific expression pattern of the candidate gene determined in transgenic lines of Arabidopsis thaliana expressing the chimeric GUS::GFP gene marker under the control of the gene promoter; gene expression response to abiotic stress and nutritional treatments quantified by Quantitative Real Time-PCR (qPCR); and phenotype of the homo