An open-field 3-year-long microplot experiment was set up with three micropropagated lines (SC Blossom, BFT Indiana, and STM Hajdúsági) of giant reed (Arundo donax L.). Plants were grown on a soil cover of a former sewage settling pond located in Debrecen Lovász-Zug, Hungary. Soil cover of the sewage sediment was moderately contaminated with various toxic elements (As, Ba, Cd, Cr, Cu, Mn, Ni, Pb, and Zn). The highest total concentration of examined toxic elements in leaves was found in the BFT Indiana line (∑326 mg/kg), while in the SC Blossom and STM Hajdúsági lines, ∑210 mg/kg and ∑182 mg/kg were measured, respectively. The highest Zn concentration (117 mg/kg) was found in the leaves of in BFT Indiana line and was 67% higher than that in SC Blossom and 95% more than in the STM Hajdúsági line. The BFT Indiana leaves showed typical signs of adaptation to heavy metal stress in the case of numerous micromorphometric characteristics. The extent of leaf mesophylls decreased, and the number of bulliform cells and phytoliths, as well as the sclerenchymatous stock, increased. The size of the vascular bundles was reduced. The size of the stomata decreased while the stomatal density increased. It can be concluded that the BFT Indiana line had the best adaptational response to heavy metal stress.

Open-field small plot long-term experiment was set up during 2011 with willow (Salix triandra × S. viminalis ‘Inger’), grown as a short rotation coppice energy crop in Nyíregyháza, Hungary. The sandy loam Cambisol with neutral pH was treated three times (2011, 2013, and 2016) with 15 t ha^–1 municipal sewage sludge compost (MSSC) and with 600 kg ha^–1 (2011, 2013) or 300 kg ha^–1 (2016) wood ash (WA). In 2018 the MSSC-treated plots were amended with 7.5 t ha^–1 municipal sewage sediment (MSS), and 300 kg ha^–1 WA. MSSC and WA or MSS and WA were also applied to the soil in combinations during all treatments. Control plots remained untreated since 2011. Repeated application of wastewater solids (MSSC, MSS) and wood ash (WA) significantly enhanced the amounts of As (up to +287%), Ba, Cd (up to +192%), Cu, Mn, Pb, and Zn in the topsoil of willows. The combined application of MSSC+MSS+WA resulted in significantly higher Mn and Zn and lower As Ba, Cd Cr, and Pb concentrations in topsoil than MSSC+MSS treatment of soil without WA. Nitrogen concentrations in leaves of treated plants were generally slightly lower or similar to control. All soil treatments significantly enhanced the uptake or accumulation of nutrient elements (Ca, K, Mg, P) and potentially toxic elements (As, Ba, Cd, Cr, Cu, Mn, Ni, Pb, and Zn) in the leaves of willows during 2018, 2019, and 2020. Significantly higher Mn or Zn concentrations were measured in MSSC+MSS+WA than in MSSC+MSS treatments. Significant amounts of Cd (up to 1.11 mg kg^–1) or Zn (up to 183 mg kg^–1) can be translocated (phytoextracted) from a soil amended with wastewater solids or wood ash to willow leaves. In 2018 the treatments decreased the chlorophyll fluorescence values, while in 2019 and 2020 the light adapted fluorescence yield (Y) values were higher in treated than in control plants.

This open-field small-plot long-term experiment was set up between 2011 and 2021 with willow (Salix triandra × S. viminalis ‘Inger’), grown as a short rotation coppice energy crop in Nyíregyháza, Hungary. The sandy loam Cambisol was treated with wastewater solids (WS) in the form of municipal sewage sludge compost (MSSC, 2011, 2013, and 2016), municipal sewage sediment (MSS, 2018), and with willow ash (WA, 2011, 2013, 2016, and 2018). Control plots remained untreated since 2011. All soil treatments significantly enhanced the uptake or accumulation of potentially toxic elements (PTEs) in the leaves of willows. During June 2019, 53 weeks after the last soil treatments, MSSC + MSS-, WA-, and MSSC + MSS + WA-treated willows leaves had 14–68% more As, 17–48% more Ba, 31–104% more Cr, 4–12% more Cu, 6–15% more Mn, 18–218% more Pb, and 11–35% more Zn compared to the untreated control. Significantly higher Mn and Zn concentrations were measured in the MSSC + MSS + WA treatments than in the MSSC + MSS treatments. The assumption that WA reduces the accumulation of PTEs in willow leaves when applied together with MSSC and MSS was therefore only partially confirmed. The hypothesis of this study was that PTEs accumulated in the leaves would affect the microanatomical parameters of the leaves. Numerous positive changes were observed with the combined application of WS and WA. MSSC + MSS + WA treatment reduced the thickness of the mesophyll less than MSSC + MSS or WA treatments alone; the size of the cells building the palisade and spongy parenchyma and the extent of the main vein significantly increased. In the case of the combined treatment, the extent of the sclerenchymatous stock was smaller than in the control but larger than in WS- or WA-treated willow. The extent of the collenchymatous stock significantly increased compared to the control. Increases in the thickness of the adaxial epidermis and the number of stomata were statistically significant. However, the extent of the increases did not reach the extent of the increase experienced in the case of WS treatment, as the size of the stomata did not significantly decrease.

A small-plot long-term field fertilization experiment was set up in 2011 with willow (Salix triandra x Salix viminalis ’Inger’) grown as an energy crop in Nyíregyháza, Hungary. The brown forest soil was treated three times (in June 2011, May 2013, May 2016) with municipal biocompost (MBC), municipal sewage sludge compost (MSSC) or willow ash (WA), and twice (June 2011, May 2013) with rhyolite tuff (RT). In late May – early June 2016 urea (U) and sulphuric urea (SU) fertilizers were also applied to the soil as top-dressing (TD). These fertilizers and amendments were also applied to the soil in 2016 in the combinations; MBC+SU, RT+SU, WA+SU and MSSC+WA. All the treatments were repeated four times. In July 2016 the highest nitrogen concentrations in willow leaves were measured in the U (3.47 m/m%) and SU (3.01 m/m%) treatments, and these values were significantly higher than the control (2.46 m/m%). An excess of nitrogen considerably reduced the Zn uptake of the leaves, with values of 39.5 μg g^-1 in the U treatment, 53.4 μg g^-1 in the SU treatment, and 63.5 μg g^-1 in the control. All other amendments or TDs, except for WA, enhanced the specific potassium concentrations in willow leaves compared to the control. No significant quantities of toxic elements (As, Ba, Cd, Pb) were transported from soil amendments or TDs to the willow leaves. In July 2016 the most intensive leaf chlorophyll fluorescence was observed in the MSSC and MSSC+WA treatments.

Willow was cultivated as an energy crop in a field experiment. The brown forest soil was treated with an inorganic fertilizer (ammonium nitrate−AN: 100 kg ha^-1) or with various organic or mineral soil amendments (municipal biocompost–MBC: 20 t ha⁻¹; municipal sewage sludge compost–MSSC: 15 t ha⁻¹; rhyolite tuff–RT: 30 t ha⁻¹; willow ash−WA: 600 kg ha⁻¹), or their combinations (AN+MBC; AN+RT; AN+WA, MSSC+WA) in four replications. Nineteen months after the soil treatments the macroelement-rich amendments (MBC, MSSC) enhanced the harvested fresh shoot yield most significantly (up to 41% as compared to the untreated control), and also the shoot diameter and shoot height of the willow plants. Most of the treatments enhanced the uptake of N (9.8-23.5%) and K in willow leaves, but the concentrations of P, Mg, Ca, Fe and Zn in the leaves were reduced. The toxic element (As, Cd, Pb) accumulation of willow shoots was negligible. In a 4-year field experiment the effects of the mineral fertilizers AN and AN+calcium-magnesium carbonate (CMC) were studied on the mineral nutrition of the leaves and wood yield of black locust trees cultivated as an energy crop. The brown forest soil was treated with 300 kg ha⁻¹ annual doses of these fertilizers as top-dressing in June 2009, May 2010 and May 2011. Both fertilizers caused a three to four times increase in the nitrate content of the upper soil soon after their application in June. By the end of the vegetation period (in December) the nitrate concentration in the soil was similar to that in the control plots. The nitrogen content of the leaf stalks (petioles) and leaves, however, was only slightly higher in the treated plots. As a trend, fertilization increased the phosphorus and reduced the calcium uptake in the leaf stalks and leaves, while the magnesium content was not influenced. In March 2012, when the whole trees were harvested, 22% or 28% higher aboveground fresh shoot weight was detected in the AN or AN+CMC treatments than in the control.