Isolation and recrystallization of epicuticular waxes from Sorbus and Cotoneaster leaves

Wax morphology and chemical composition are widely accepted to be important for the protective properties of the leaf’s surface and also valuable characteristics in plant systematics. The leaves of Sorbus domestica L. and Cotoneaster granatensis Boiss., species of two large genera with intricate taxonomy referred to subtribe Pyrinae, Rosaceae (formerly subfamily Maloideae), were studied by scanning electron microscope (SEM) and performing different methods of wax isolation. The aim of the study was to acquire a suitable, cost and time effective method for wax removal. Chloroform and methanol extractions and freeze-embedding method for direct isolation of the wax crystals were applied. Immersing the leaves for 3 minutes in chloroform was sufficient to extract the waxes whereas the efficiency of the methanol solvent was lower. Wax layers with wellpreserved structures of the crystals from both upper and lower epidermis were successfully transferred to artificial surfaces. The recrystallization experiment demonstrated that waxes from chloroform extracts could recrystallize in in vitro conditions on artificial surfaces. The crystals showed same micromorphology as on the intact leaves. Results of this study could be applied in further analytical researches of the waxes of S. domestica and C. granatensis or other species of the subtribe Pyrinae

Seed Priming with Single-Walled Carbon Nanotubes Grafted with Pluronic P85 Preserves the Functional and Structural Characteristics of Pea Plants

The engineering of carbon nanotubes in the last decades resulted in a variety of applications in electronics, electrochemistry, and biomedicine. A number of reports also evidenced their valuable application in agriculture as plant growth regulators and nanocarriers. In this work, we explored the effect of seed priming with single-walled carbon nanotubes grafted with Pluronic P85 polymer (denoted P85-SWCNT) on Pisum sativum (var. RAN-1) seed germination, early stages of plant development, leaf anatomy, and photosynthetic efficiency. We evaluated the observed effects in relation to hydro- (control) and P85-primed seeds. Our data clearly revealed that seed priming with P85-SWCNT is safe for the plant since it does not impair the seed germination, plant development, leaf anatomy, biomass, and photosynthetic activity, and even increases the amount of photochemically active photosystem II centers in a concentration-dependent manner. Only 300 mg/L concentration exerts an adverse effect on those parameters. The P85 polymer, however, was found to exhibit a number of negative effects on plant growth (i.e., root length, leaf anatomy, biomass accumulation and photoprotection capability), most probably related to the unfavorable interaction of P85 unimers with plant membranes. Our findings substantiate the future exploration and exploitation of P85-SWCNT as nanocarriers of specific substances promoting not only plant growth at optimal conditions but also better plant performance under a variety of environmental stresses

Single-walled carbon nanotubes modify leaf micromorphology, chloroplast ultrastructure and photosynthetic activity of pea plants

Single-walled carbon nanotubes (SWCNTs) emerge as promising novel carbon-based na-noparticles for use in biomedicine, pharmacology and precision agriculture. They were shown to penetrate cell walls and membranes and to physically interact and exchange electrons with photosynthetic complexes in vitro. Here, for the first time, we studied the concentration-dependent effect of foliar application of copolymer-grafted SWCNTs on the structural and functional characteristics of intact pea plants. The lowest used concentration of 10 mg L−1 did not cause any harmful effects on the studied leaf characteristics, while abundant epicuticular wax generation on both leaf surfaces was observed after 300 mg L−1 treatment. Swelling of both the granal and the stromal regions of thylakoid membranes was detected after application of 100 mg L−1 and was most pronounced after 300 mg L−1. Higher SWCNT doses lead to impaired photosynthesis in terms of lower proton motive force generation, slower generation of non-photochemical quenching and reduced zeaxanthin con-tent; however, the photosystem II function was largely preserved. Our results clearly indicate that SWCNTs affect the photosynthetic apparatus in a concentration-dependent manner. Low doses (10 mg L−1) of SWCNTs appear to be a safe suitable object for future development of nanocarriers for substances that are beneficial for plant growth. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Physiological and structural adjustments of two ecotypes of Platanus orientalis L. from different habitats in response to drought and re-watering

Platanus orientalis covers a very fragmented area in Europe and, at the edge of its natural distribution, is considered a relic endangered species near extinction. In our study, it was hypothesized that individuals from the edge of the habitat, with stronger climate constrains (drier and warmer environment, Italy, IT ecotype), developed different mechanisms of adaptation than those growing under optimal conditions at the center of the habitat (more humid and colder environment, Bulgaria, BG ecotype). Indeed, the two P. orientalis ecotypes displayed physiological, structural and functional differences already under control (unstressed) conditions. Adaptation to a dry environment stimulated constitutive isoprene emission, determined active stomatal behavior, and modified chloroplast ultrastructure, ultimately allowing more effective use of absorbed light energy for photochemistry. When exposed to short-term acute drought stress, IT plants showed active stomatal control that enhanced instantaneous water use efficiency, and stimulation of isoprene emission that sustained photochemistry and reduced oxidative damages to membranes, as compared to BG plants. None of the P. orientalis ecotypes recovered completely from drought stress after re-watering, confirming the sensitivity of this mesophyte to drought. Nevertheless, the IT ecotype showed less damage and better stability at the level of chloroplast membrane parameters when compared to the BG ecotype, which we interpret as possible adaptation to hostile environments and improved capacity to cope with future, likely more recurrent, drought stress