Dried leaf samples were ground using 4 mm diameter inox beads in an oscillating grinder (Mixer Mill MM400, Retsch, Haan, Germany). The concentrations of most elements (Mg, P, S, K, Ca, B, Mn, Fe, Ni, Cu, Zn, Mo Na, Co, V, and Se) were quantified using the procedure previously described [102 (link)] using 40 mg of dry powder per sample that was submitted to acid digestion and mineralization. The resulting solutions were then analyzed by high-resolution inductively coupled plasma mass spectrometry (HR-ICP-MS, Element 2TM, Thermo Fisher Scientific, Bremen, Germany).
The total N concentration was determined with 1.5 mg of fine powder placed in tin capsules before analysis with an isotope-ratio mass spectrometer (IRMS, Isoprime, GV Instruments, Manchester, UK) linked to a C/N/S analyzer (EA3000, Euro Vector, Milan, Italy).
The remaining elements (Cl, Si, and Al) were quantified with approximately 1 g of dry weight powder analyzed with an X-ray-fluorescence spectrometer (XEPOS, Ametek, Berwyn, PA, USA) using calibration curves obtained from international standards with known concentrations.
In this study, elemental content is expressed as the ratio WD/control plants following the calculation method described [61 (link)].
The total N concentration was determined with 1.5 mg of fine powder placed in tin capsules before analysis with an isotope-ratio mass spectrometer (IRMS, Isoprime, GV Instruments, Manchester, UK) linked to a C/N/S analyzer (EA3000, Euro Vector, Milan, Italy).
The remaining elements (Cl, Si, and Al) were quantified with approximately 1 g of dry weight powder analyzed with an X-ray-fluorescence spectrometer (XEPOS, Ametek, Berwyn, PA, USA) using calibration curves obtained from international standards with known concentrations.
In this study, elemental content is expressed as the ratio WD/control plants following the calculation method described [61 (link)].
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