A digital caliper was used for measuring the length of the root and shoot. The dry weight of root and shoot was recorded after drying for 3 days at 80 °C in an electric oven. For measuring the chlorophyll score of young leaves, a SPAD meter was used (Minolta, Japan). Furthermore, photosynthesis biophysics through chlorophyll fluorescence kinetic (OJIP), such as maximum quantum yield of PSII (FvFm), photosynthesis performance index (Pi_ABS), dissipation energy per active reaction center (DIo/RC), absorption flux/effective antenna size of an active reaction center (ABS/RC), electron transporter flux further than QA DIo/RC (ET2o/RC) and trapped energy flux leading to a reduction of QA (TRo/RC) was recorded on young leaves kept for 1 h at dark using FluorPen FP 100 (Photon Systems Instruments, Czech Republic).
Fe chelate reductase activity (FCR) activity in roots was determined in roots by ferrozine assay15 (link). Briefly, the 0.2 mM CaSO4 followed by Milli-Q water washed off the root surface. The roots samples were then homogenized with 1 ml of assay mixture (100 mM Fe(III) EDTA, 0.10 mmol MES-NaOH (pH 5.5), 300 mM ferrozine). The samples and blank tubes (without assay mixture) were incubated in the dark for 20 min at 25 °C. Finally, aliquots were read at 562 nm. The FCR activity was determined by using the molar extinction coefficient (27.9 mM−1 cm−1) of ferrozine.
Fe chelate reductase activity (FCR) activity in roots was determined in roots by ferrozine assay15 (link). Briefly, the 0.2 mM CaSO4 followed by Milli-Q water washed off the root surface. The roots samples were then homogenized with 1 ml of assay mixture (100 mM Fe(III) EDTA, 0.10 mmol MES-NaOH (pH 5.5), 300 mM ferrozine). The samples and blank tubes (without assay mixture) were incubated in the dark for 20 min at 25 °C. Finally, aliquots were read at 562 nm. The FCR activity was determined by using the molar extinction coefficient (27.9 mM−1 cm−1) of ferrozine.