The Influence of Glow and Afterglow Cold Plasma Treatment on Biochemistry, Morphology, and Physiology of Wheat Seeds.

Cold plasma (CP) technology is a technique used to change chemical and morphological characteristics of the surface of various materials. It is a newly emerging technology in agriculture used for seed treatment with the potential of improving seed germination and yield of crops. Wheat seeds were treated with glow (direct) or afterglow (indirect) low-pressure radio-frequency oxygen plasma. Chemical characteristics of the seed surface were evaluated by XPS and FTIR analysis, changes in the morphology of the seed pericarp were analysed by SEM and AFM, and physiological characteristics of the seedlings were determined by germination tests, growth studies, and the evaluation of α-amylase activity. Changes in seed wettability were also studied, mainly in correlation with functionalization of the seed surface and oxidation of lipid molecules. Only prolonged direct CP treatment resulted in altered morphology of the seed pericarp and increased its roughness. The degree of functionalization is more evident in direct compared to indirect CP treatment. CP treatment slowed the germination of seedlings, decreased the activity of α-amylase in seeds after imbibition, and affected the root system of seedlings.

Use of Plasma Technologies for Antibacterial Surface Properties of Metals.

Bacterial infections of medical devices present severe problems connected with long-term antibiotic treatment, implant failure, and high hospital costs. Therefore, there are enormous demands for innovative techniques which would improve the surface properties of implantable materials. Plasma technologies present one of the compelling ways to improve metal's antibacterial activity; plasma treatment can significantly alter metal surfaces' physicochemical properties, such as surface chemistry, roughness, wettability, surface charge, and crystallinity, which all play an important role in the biological response of medical materials. Herein, the most common plasma treatment techniques like plasma spraying, plasma immersion ion implantation, plasma vapor deposition, and plasma electrolytic oxidation as well as novel approaches based on gaseous plasma treatment of surfaces are gathered and presented. The latest results of different surface modification approaches and their influence on metals' antibacterial surface properties are presented and critically discussed. The mechanisms involved in bactericidal effects of plasma-treated surfaces are discussed and novel results of surface modification of metal materials by highly reactive oxygen plasma are presented.

The Combined Inhibition of Autophagy and Diacylglycerol Acyltransferase-Mediated Lipid Droplet Biogenesis Induces Cancer Cell Death during Acute Amino Acid Starvation.

Lipid droplets (LDs) are dynamic organelles involved in the management of fatty acid trafficking and metabolism. Recent studies suggest that autophagy and LDs serve complementary roles in the protection against nutrient stress, but the autophagy-LD interplay in cancer cells is not well understood. Here, we examined the relationship between autophagy and LDs in starving HeLa cervical cancer- and MDA-MB-231 breast cancer cells. We found that acute amino acid depletion induces autophagy and promotes diacylglycerol acyltransferase 1 (DGAT1)-mediated LD accumulation in HeLa cells. Inhibition of autophagy via late-stage autophagy inhibitors, or by knocking down autophagy-related 5 (ATG5), reduced LD accumulation in amino acid-starved cancer cells, suggesting that autophagy contributes to LD biogenesis. On the contrary, knockdown of adipose triglyceride lipase (ATGL) increased LD accumulation, suggesting that LD breakdown is mediated by lipolysis under these conditions. Concurrent inhibition of autophagy by silencing ATG5 and of LD biogenesis using DGAT inhibitors was effective in killing starving HeLa cells, whereas cell survival was not compromised by suppression of ATGL-mediated lipolysis. Autophagy-dependent LD biogenesis was also observed in the aggressive triple-negative MDA-MB-231 breast cancer cells deprived of amino acids, but these cells were not sensitized to starvation by the combined inhibition of LD biogenesis and autophagy. These findings reveal that while targeting autophagy-driven and DGAT-mediated LD biogenesis reduces the resilience of HeLa cervical cancer cells to amino acid deprivation, this strategy may not be successful in other cancer cell types.

Chemical alterations of grain surface by cold plasma technology: Comparison of buckwheat and wheat grain responses to oxygen low-pressure plasma.

Cold plasma (CP) has a great potential for decontamination or improvement of grain germination. However, disputing results have been reported, as plasma treatment can affect species and varieties of grains in different ways. The differences may be due to the chemical composition of grain pericarps, the structure of the grains and metabolic response mechanisms. CP treatment decreased grain germination rate, speed and activity of α-amylase of buckwheat grains. Such effects on both varieties of wheat grains were present after longer exposure to plasma. Lipid peroxidation was highest in buckwheat grains, whereas wheat grains were less affected. Plasma-treated Gorolka variety exhibited a low level of lipid peroxidation, no different to untreated grains, compared to Primorka grains, where longer treatment triggered higher levels of lipid peroxidation. The response of grains to CP treatment depends on the chemical and structural properties of grains pericarp, as well as plant tolerance to certain abiotic conditions.

Gold-Assisted Molecular Imaging of Organic Tissue by MeV Secondary Ion Mass Spectrometry.

The quality of molecular imaging by means of MeV primary ion-induced secondary ion mass spectrometry by coating with gold was evaluated on different reference organic molecules and plant samples. The enhancement of the secondary ion yield was evident for the majority of the studied analytes, reaching the highest values at gold thicknesses between 0.5 and 2 nm, and increased the intensity up to 5-fold for reference samples and >2-fold for specific peaks within the plant sample. Improved propagation of the electric field due to the target potential on otherwise electrically insulating plant samples was also evident through improved image resolution and by reducing the background in mass spectra. However, detection of several molecules was significantly decreased at even at 1 nm thick gold layer. The results indicated that an optimized sequence of analysis is required to reliably interpret results.

The Sensitivity of Fungi Colonising Buckwheat Grains to Cold Plasma Is Species Specific.

Fungi are the leading cause of plant diseases worldwide and are responsible for enormous agricultural and industrial losses on a global scale. Cold plasma (CP) is a potential tool for eliminating or inactivating fungal contaminants from biological material such as seeds and grains. This study used a low-pressure radiofrequency CP system with oxygen as the feed gas to test the decontamination efficacy of different genera and species commonly colonising buckwheat grains. Two widely accepted methods for evaluating fungal decontamination after CP treatment of seeds were compared: direct cultivation technique or contamination rate method (%) and indirect cultivation or colony-forming units (CFU) method. For most of the tested fungal taxa, an efficient decrease in contamination levels with increasing CP treatment time was observed. Fusarium graminearum was the most susceptible to CP treatment, while Fusarium fujikuroi seems to be the most resistant. The observed doses of oxygen atoms needed for 1-log reduction range from 1024-1025 m-2. Although there was some minor discrepancy between the results obtained from both tested methods (especially in the case of Fusarium spp.), the trends were similar. The results indicate that the main factors affecting decontamination efficiency are spore shape, size, and colouration.

Exploring the potential of cold plasma treatment followed by zinc-priming for biofortification of buckwheat sprouts.

Increasing the concentration of an element in edible produce (i.e., biofortification) can mitigate the element deficiency in humans. Sprouts are small but popular part of healthy diets providing vitamins and essential elements throughout the year. Element composition of sprouts can easily be amended, e.g., by soaking the grains in element-rich solution before germination (grain-priming). In addition, pre-treatment of grains to improve element translocation from the solution into the grain may further enhance the element concentration in the sprout. Cold plasma technique could provide such solution, as it increases wettability and water uptake of grains. Grains of common buckwheat (Fogopyrum esculentum Moench) were pre-treated/ untreated with cold plasma and soaked in ZnCl2 solution/pure water. Germination tests, α-amylase activity, grain hydrophilic properties and water uptake were assessed. Element composition of grain tissues and of sprouts was assessed by micro-particle-induced-X-ray emission and X-ray fluorescence spectroscopy, respectively. Grain-priming increased Zn concentration in shoots of common buckwheat sprouts more than five-times, namely from 79 to 423 mg Zn kg-1 dry weight. Cold plasma treatment increased grain wettability and water uptake into the grain. However, cold plasma pre-treatment followed by grain-priming with ZnCl2 did not increase Zn concentration in different grain tissues or in the sprouts more than the priming alone, but rather decreased the Zn concentration in sprout shoots (average ± standard error: 216 ± 6.13 and 174 ± 7.57 mg Zn kg-1 dry weight, respectively). When the fresh weight portion of whole sprouts (i.e., of roots and shoots) was considered, comparable average requirements of Zn, namely 24.5 % and 35 % for adult men and women would be satisfied by consuming cold plasma pre-treated and not pre-treated grains. Potential advantages of cold plasma pre-treatment need to be tested further, mainly to optimize the duration of soaking required to produce Zn-enriched sprouts.

Decontamination and Germination of Buckwheat Grains upon Treatment with Oxygen Plasma Glow and Afterglow.

Buckwheat is an alternative crop known for its many beneficial effects on our health. Fungi are an important cause of plant diseases and food spoilage, often posing a threat to humans and animals. This study reports the effects of low-pressure cold plasma treatment on decontamination and germination of common (CB) and Tartary buckwheat (TB) grains. Both plasma glow and afterglow were applied. The glow treatment was more effective in decontamination: initial contamination was reduced to less than 30% in CB and 10% in TB. Fungal diversity was also affected as only a few genera persisted after the glow treatment; however, it also significantly reduced or even ceased the germination capacity of both buckwheat species. Detailed plasma characterisation by optical spectroscopy revealed extensive etching of outer layers as well as cotyledons. Afterglow treatment resulted in a lower reduction of initial fungal contamination (up to 30% in CB and up to 50% in TB) and had less impact on fungal diversity but did not drastically affect germination: 60-75% of grains still germinated even after few minutes of treatment. The vacuum conditions alone did not affect the fungal population or the germination despite an extensive release of water.

Response of Two Different Wheat Varieties to Glow and Afterglow Oxygen Plasma.

Cold plasma technology has received significant attention in agriculture due to its effect on the seeds and plants of important cultivars, such as wheat. Due to climate change, wherein increasing temperatures and droughts are frequent, it is important to consider novel approaches to agricultural production. As increased dormancy levels in wheat are correlated with high temperatures and drought, improving the germination and root growth of wheat seeds could offer new possibilities for seed sowing. The main objective of this study was to evaluate the influence of direct (glow) and indirect (afterglow) radio-frequency (RF) oxygen plasma treatments on the germination of two winter wheat varieties: Apache and Bezostaya 1. The influence of plasma treatment on seed surface morphology was studied using scanning electron microscopy, and it was observed that direct plasma treatment resulted in a high etching and nanostructuring of the seed surface. The effect of plasma treatment on germination was evaluated by measuring the germination rate, counting the number of roots and the length of the root system, and the fresh weight of seedlings. The results of this study indicate that the response of seeds to direct and indirect plasma treatment may be variety-dependent, as differences between the two wheat varieties were observed.

Cold Plasma Affects Germination and Fungal Community Structure of Buckwheat Seeds.

Crop seeds are frequently colonised by fungi from the field or storage places. Some fungi can cause plant diseases or produce mycotoxins, compromising the use of seeds as seeding material, food or feed. We have investigated the effects of cold plasma (CP) on seed germination and diversity of seed-borne fungi in common and Tartary buckwheat. The seeds were treated with CP for 15, 30, 45, 60, 90, and 120 s in a low-pressure radiofrequency system using oxygen as the feed gas. The fungi from the seed surface and fungal endophytes were isolated using potato dextrose agar plates. After identification by molecular methods, the frequency and diversity of fungal strains were compared between CP treated and chemically surface-sterilised (30% of H2O2) seeds. CP treatments above 60 s negatively affected the germination of both buckwheat species. A significant reduction in fungal frequency and diversity was observed after 90 s and 120 s in common and Tartary buckwheat, respectively. The filamentous fungi of genera Alternaria and Epicoccum proved to be the most resistant to CP. The results of our study indicate that CP treatment used in our study may be applicable in postharvest and food production, but not for further seed sowing.

Effects of Nonthermal Plasma on Morphology, Genetics and Physiology of Seeds: A Review.

Nonthermal plasma (NTP), or cold plasma, has shown many advantages in the agriculture sector as it enables removal of pesticides and contaminants from the seed surface, increases shelf life of crops, improves germination and resistance to abiotic stress. Recent studies show that plasma treatment indeed offers unique and environmentally friendly processing of different seeds, such as wheat, beans, corn, soybeans, barley, peanuts, rice and Arabidopsis thaliana, which could reduce the use of agricultural chemicals and has a high potential in ecological farming. This review covers the main concepts and underlying principles of plasma treatment techniques and their interaction with seeds. Different plasma generation methods and setups are presented and the influence of plasma treatment on DNA damage, gene expression, enzymatic activity, morphological and chemical changes, germination and resistance to stress, is explained. Important plasma treatment parameters and interactions of plasma species with the seed surface are presented and critically discussed in correlation with recent advances in this field. Although plasma agriculture is a relatively new field of research, and the complex mechanisms of interactions are not fully understood, it holds great promise for the future. This overview aims to present the advantages and limitations of different nonthermal plasma setups and discuss their possible future applications.