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Despite extensive study of extracellular vesicles (EVs), specifically exosomes (EXs) as biomarkers, important modulators of physiological or pathological processes, or therapeutic agents, relatively little is known about nonconventional sources of EXs, such as invertebrate or plant EXs, and their uses. Likewise, there is no clear information on the overview of storage conditions and currently used isolation methods, including new ones, such as microfluidics, which fundamentally affect the characterization of EXs and their other biomedical applications. The purpose of this review is to briefly summarize conventional and nonconventional sources of EXs, storage conditions and typical isolation methods, widely used kits and new “smart” technologies with emphasis on the influence of isolation techniques on EX content, protein detection, RNA, mRNA and others. At the same time, attention is paid to a brief overview of the direction of biomedical application of EXs, especially in diagnostics, therapy, senescence and aging and, with regard to the current situation, in issues related to Covid-19.

Second-generation biofuel crop miscanthus is one of the most promising plants tested for phytomanagement of contaminated sites. In this preliminary pot case study, the most used hybrid Miscanthus x giganteus was cultivated in three different real contaminated soils: agricultural soil contaminated with Cd; post-military soil slightly contaminated with Zn, Pb and Cd; and soil contaminated by petroleum industry with metals and hydrocarbons. The stress response of plants and soil microbial communities was monitored to receive data that are important for successful phytomanagement application. With metals only, the plant grew well, and chlorophyll fluorescence measurement proved their good vitality. Changes in leaf anatomy (leaf thickness and sclerenchyma cells area) were additionally determined in post-military soil compared to agricultural. On the contrary, in petroleum-contaminated soil, the biomass yield was too reduced and also physiological parameters were significantly decreased. The response of microbial communities also differed. In agricultural soil, no microbial stress was determined. In post-military soil, it became reduced during the experiment, and in petroleum contamination, it increased year-on-year. It could be concluded that miscanthus is suitable for cultivation in metals contaminated soils with potential for microbial communities support, but in soil contaminated by the petroleum industry, its application did not seem meaningful.

Bioenergy crops such as Miscanthus × giganteus are foreseeable as an alternative source to replace fossil fuel and reduce greenhouse gas emissions. They are also assessed as an environment-friendly solution for polluted, marginal and low-quality agricultural soils. Several studies had been launched on soil organic carbon sequestration potentials of miscanthus culture along with its impacts on restoring soil functionality, most of which focus on the long-term basis of the plant’s cultivation. Nevertheless, information concerning the short term impacts as well as the situation in Czechia is still scarce. In this context, a field experiment was launched in 2017 in a poor-quality agricultural land in the city of Chomutov (North-Western Czechia) to compare the impacts of the perennial C4 miscanthus with an annual C3 forage crop (wheat) on the soil carbon stocks as well as enhancing its functionality. Results through the 0–30 cm soil profile examination showed that miscanthus plants played a role in improving the studied soil physico-chemical (bulk density and soil organic carbon concentrations) and biological (Phospholipid fatty acids stress indicator, basal respiration and fluorescein diacetate hydrolytic activity) parameters. The naturally occurring δ13C concentrations were used to evaluate the direct plant contribution to the total soil organic carbon (SOC) stocks and revealed considerable miscanthus contribution all over the detected soil layers (1.98 ± 0.21 Mg C. ha−1 yr−1) after only 3 growing seasons. It is thus suggested that the C4 perennial miscanthus possess remarkable prospects for SOC sequestration and restoring degraded lands.

Abstract Plant extracellular vesicles (pEVs) derived from numerous edible sources gain a lot of attention in recent years, mainly due to the potential to efficiently carry bioactive molecules into mammalian cells. In the present study, we focus on isolation of PDNVs (plant-derived nanovesicles) and pEVs from callus culture and from BY-2 culture of Nicotiana tabacum (tobacco). Tobacco was selected as a source of plant vesicles, as it is commonly used by human, moreover it is a model organism with established techniques for cultivation of explant cultures in vitro. Explant cultures are suitable for the isolation of pEVs in large quantities, due to their fast growth in sterile conditions. As the efficiency of isolation methods varies, we were comparing two methods of isolation. We evaluated biophysical and biochemical properties of plant vesicles, as well as differences between isolates. We encountered difficulties in the form of vesicles aggregation, which is often described in publications focused on mammalian nanovesicles. In an effort to prevent vesicle aggregation, we used trehalose in different stages of isolation. We show tobacco-derived vesicles successfully enter tobacco and mesenchymal cell lines. We observed that tobacco-nanovesicles isolated by different methods incorporated fluorescent dye with different efficiency. The results of our study show tobacco-derived vesicles isolated by various isolation methods are able to enter plant, as well as mammalian cells.