Microanatomical Properties of Energy Willow (Salix spp.) Leaves after Exposure to Potentially Toxic Elements from Wastewater Solids and Wood Ash

Abstract

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.