An improved method for the visualization of conductive vessels in Arabidopsis thaliana inflorescence stems

Dye perfusion is commonly used for the identification of conductive elements important for the study of xylem development as well as precise hydraulic estimations. The tiny size of inflorescence stems, the small amount of vessels in close arrangement, and high hydraulic resistivity delimit the use of the method for quantification of the water conductivity of Arabidopsis thaliana, one of the recently most extensively used plant models. Here, we present an extensive adjustment to the method in order to reliably identify individual functional (conductive) vessels. Segments of inflorescence stems were sealed in silicone tubes to prevent damage and perfused with a dye solution. Our results showed that dyes often used for staining functional xylem elements (safranin, fuchsine, toluidine blue) failed with Arabidopsis. In contrast, Fluorescent Brightener 28 dye solution perfused through segments stained secondary cell walls of functional vessels, which were clearly distinguishable in native cross sections. When compared to identification based on the degree of development of secondary cell walls, identification with the help of dye perfusion revealed a significantly lower number of functional vessels and values of theoretical hydraulic conductivity. We found that lignified but not yet functional vessels form a substantial portion of the xylem in apical and basal segments of Arabidopsis and, thus, significantly affect the analyzed functional parameters of xylem. The presented methodology enables reliable identification of individual functional vessels, allowing thus estimations of hydraulic conductivities to be improved, size distributions and vessel diameters to be refined, and data variability generally to be reduced

Copper demand, supply, and associated energy use to 2050

To a set of well-regarded international scenarios (UNEP's GEO-4), we have added consideration of the demand, supply, and energy implications related to copper production and use over the period 2010-2050. To our knowledge, these are the first comprehensive metal supply and demand scenarios to be developed. We find that copper demand increases by between 275 and 350% by 2050, depending on the scenario. The scenario with the highest prospective demand is not Market First (a "business as usual" vision), but Equitability First, a scenario of transition to a world of more equitable values and institutions. These copper demands exceed projected copper mineral resources by mid-century and thereafter. Energy demand for copper production also demonstrates strong increases, rising to as much as 2.4% of projected 2050 overall global energy demand. We investigate possible policy responses to these results, concluding that improving the efficiency of the copper cycle and encouraging the development of copper-free energy distribution on the demand side, and improving copper recycling rates on the supply side are the most promising of the possible options. Improving energy efficiency in primary copper production would lead to a reduction in the energy demand by 0.5% of projected 2050 overall global energy demand. In addition, encouraging the shift towards renewable technologies is important to minimize the impacts associated with copper production