Aa500fg

Seedlings were grown over 1.5 months and then seedling growth was recorded in six seedlings per treatment. The number of leaves and seedling height were measured. Leaf stomatal conductance was measured by using a ∆T-Porometer AP4 (Delta-T Devices-Cambridge, UK). Leaf chlorophyll fluorescence (chlorophyll fluorometer, opti-sciences OS-30p, UK) was measured on two fully expanded leaves per plant, along with leaf chlorophyll content (six replications/treatment) [35] (link). Plants were harvested above substrate, upper fresh weight was weighed (g), dried, and total dry matter content (%) was then computed.
Mineral accumulation in upper part of the plant (including leaves and shoots) was measured on three replications/treatment (two pooled plants/replication) as described previously [36] (link). The content of nitrogen (N) was determined with Kjeldahl digestion method (BUCHI, Digest automat K-439 and Distillation Kjeldahl K-360, Oldham, UK), while phosphorus (P) spectrophotometrically (Multiskan GO, Thermo Fischer Scientific, USA), and potassium (K), magnesium (Mg), calcium (Ca), sodium (Na), iron (Fe), copper (Cu), manganese (Mn), and zinc (Zn) were determined by an atomic absorption spectrometer (PG Instruments AA500FG, Leicestershire, UK). Results were indicated in g kg -1 and mg kg -1 of dry weight, for macro-and micronutrients, respectively.

The nutrient content in leaves was determined at the flowering and at the veraison phenological stage. Approximately 100 leaf stems (four replicates/treatment) were collected, dried at 65 • C for four days, weighed, and ground in a Wiley mill to pass through 40 mesh screens, as described in Marinou et al. [41] (link). Nitrogen (N) content was determined by the Kjeldahl method (BUCHI, Digest automat K-439 and Distillation Kjelflex K-360, Switzerland). Potassium (K) and sodium (Na) were determined photometrically (Flame photometer, Lasany Model 1832, Lasany International, India), phosphorus (P) was determined spectrophotometrically (Multiskan GO, Thermo Fisher Scientific, Vantaa, Finland) and magnesium (Mg) by an atomic absorption spectrophotometer (PG Instruments AA500FG, Leicestershire, UK). Data were expressed in g kg -1 of dry weight.