The mutagenic effect of cold plasma on Medicago sativa L.

To explore the feasibility of using cold plasma as a mutagenesis breeding technology for forage crops, in this study we used the Medicago sativa L. cultivar, Zhongcao No. 3, as the experimental material. The effects of plasma treatments on Medicago sativa L. were analyzed through the use of plasma and activated water. Treatments with plasma and activated water inhibited plant height but promoted root growth. By creating a closed environment, adding a dielectric barrier plate, and combining these two treatment methods, the greatest impact can be had on the growth of Medicago sativa L. seeds. After treatment, the plant heights were approximately half those of the control group, and the root lengths were approximately 1.6 times those of the control group. Through emission spectroscopy, it was found that active particles such as O, NO2, and N2* were present and could be considered to have produced plasma-activated water through contact with the water surface, thus affecting the survival and growth of the seeds. Whole-genome resequencing (WGRS) was performed on the wild-type and selected mutants after treatment, with an average sequencing depth of 115.93×, an average genome alignment rate of 91.72 %, and an average genome coverage rate of 91.85 %. Various types of mutations were detected and annotated. After filtering, 7,822,324 SNP (single nucleotide polymorphisms) sites, 2,161,917 indel sites, 200,544 SV sites, 238 CNV (copy number variation) sites. The SNPs, indels (insertions/deletions), and SVs (structural variations) were mainly heterozygous, with heterozygosity rates of 87.13 %, 92.16 %, and 83.49 %, respectively. The CNVs were dominated by low copy numbers, accounting for 53.77 %. These results indicate that plasma treatment has significant effects on plant growth and genome of Medicago sativa L.

Effect of Plasma-Activated Water on the Cellulase-Producing Strain Aspergillus niger A32.

To investigate the effect and mechanism of plasma-activated water (PAW) on Aspergillus niger, PAW was prepared using a needle array-plate dielectric barrier discharge plasma system. The concentrations of long-lived reactive oxygen and nitrogen species (RONS), namely, H2O2, NO2-, and NO3-, in the PAW were 48.76 mg/L, 0.046 mg/L, and 172.36 mg/L, respectively. Chemically activated water (CAW) with the same concentration of long-lived RONS was also prepared for comparison. A. niger A32 was treated with PAW and CAW. After treatment, the treated strains were observed and analyzed with scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to screen probable mutants. The results indicated that the pH, conductivity, and ORP values of PAW were 2.42, 1935 µS/cm, and 517.07 mV, respectively. In contrast, the pH and ORP values of CAW were 6.15 and 301.73 mV, respectively, which differed significantly from those of PAW. In addition, the conductivity of CAW showed no change. SEM and TEM analyses revealed that A. niger A32 treated with CAW exhibited less damage compared with the control. In contrast, A. niger A32 treated with PAW showed significant shrinkage, deformation, and exudate attachment over time. Following PAW treatment, after four passages, a high cellulase-producing stable mutant strain A-WW5 was screened, exhibiting a filter paper enzyme activity of 29.66 U/mL, a cellulose endonuclease activity of 13.79 U/mL, and a ß-glucosidase activity of 27.13 U/mL. These values were found to be 33%, 38%, and 2.1% higher than those of the original fungus sample, respectively. In total, 116 SNPs and 61 InDels were present in the genome of the mutant strain A-WW5. The above findings indicate that the impact of PAW on A. niger is not only attributed to long-lasting H2O2, NO2-, and NO3- particles but also to other short-lived active particles; PAW is expected to become a new microbial breeding mutagen.

Effects of discharge plasma on seed germination and volatile compounds content of Agropyron Mongolicum.

To investigate the effects of discharge plasma on Agropyron mongolicum seeds, various treatments including direct exposure to discharge plasma, combined treatment with discharge plasma and plasma-activated water (PAW) were applied to the seeds. The changes in germination rate, MDA content, and volatile compound levels of Agropyron mongolicum seeds after different treatments were examined. The results showed that the direct effect of plasma had no significant effect on the MDA content or germination rate of Agropyron mongolicum seeds due to the limited penetration depth. However, the combined effect of plasma and activated water could cause active nitrogen and oxygen particles to enter the seeds and cause oxidative stress damage. After 18 h of combined treatment, the MDA content increased significantly, and the germination rate decreased to below the semilethal dose, which was 33.44 %. After plasma treatment, 55 volatile compounds, mainly alcohols, aldehydes and ketones, were identified from the seeds of Agropyron mongolicum. Due to the oxidation and modification of the plasma, the content of most aldehydes increased with increasing reaction time. After screening, 13 volatile organic compounds could be used as potential markers to distinguish between different treatment methods. These results reveal the mechanism underlying the biological effects of plasma treatment on Agropyron mongolicum seeds.

Insight into the surface discharge cold plasma efficient inactivation of Pseudomonas fluorescens in water based on exogenous reactive oxygen and nitrogen species: Synergistic mechanism and energy benefits.

As well known, surface discharge cold plasma has efficient inactivation ability and a variety of RONS are main active particles for inactivation, but their synergistic mechanism is still not clear. Therefore, surface discharge cold plasma system was applied to treat Pseudomonas fluorescens to study bacterial inactivation mechanism and energy benefit. Results showed that energy efficiency was directly proportional to applied voltage and inversely proportional to initial concentration. Cold plasma treatment for 20 min was inactivated by approximately > 4-log10Pseudomonas fluorescens and application of •OH and 1O2 scavengers significantly improved survival rate. In addition, •OH and 1O2 destroyed cell membrane structure and membrane permeability, which promoted diffusion of RONS into cells and affecting energy metabolism and antioxidant capacity, leading to bacterial inactivation. Furthermore, accumulation of intracellular NO and ONOOH was related to infiltration of exogenous RNS, while accumulation of •OH, H2O2, 1O2, O2- was the result of joint action of endogenous and exogenous ROS. Transcriptome analysis revealed that different RONS of cold plasma were responsible for Pseudomonas fluorescens inactivation and related to activation of intracellular antioxidant defense system and regulation of genes expression related to amino acid metabolism and energy metabolism, which promoting cellular process, catalytic activity and other biochemical pathways.

Effect of electrohydrodynamic (EHD) drying on active ingredients, textural properties and moisture distribution of yam (Dioscorea opposita).

This paper systematically investigates the changes in material properties during electrohydrodynamic (EHD) drying, the discharge characteristics of the EHD system as well as the active ingredients, textural properties (hardness, adhesiveness, etc.) and moisture distribution of yam under EHD, air drying and hot air drying were investigated. The results showed that the active particles and the ionized wind generated during the discharge process of the electrohydrodynamic drying device had a significant effect on the drying. Compared to thermal drying, 21 kV drying resulted in the most complete cellular structure, the best internal bound water content as well as textural properties of yam. It played a positive role in the retention of internal nutrients in yam, and the total phenol and allantoin contents were increased by 25.74% and 81.99%, respectively. These results elucidate the advantages of electrohydrodynamic drying in yam drying and provide a reference for the application of EHD in drying.

Study on the thawing characteristics of beef in ultrasound combined with plasma-activated water.

The effects of deionized water thawing (DT), plasma-activated water thawing (PT), ultrasound (150 W, 40 kHz) combined with deionized water thawing (UDT), and ultrasound combined with plasma-activated water thawing (UPT) on the thawing characteristics and the physicochemical properties of the beef were investigated. The results showed that the UPT group had a faster thawing rate (38 % higher compared to the PT group) and good bactericidal ability (75 % higher compared to the UDT group), and had no adverse effect on the color and pH value of the beef. Plasma-activated water (PAW) can maintain the stability of the beef fiber, improve the water holding capacity (WHC), inhibit lipid oxidation, and reduce the loss of soluble substances such as protein. Therefore, UPT thawing is a promising meat thawing technology, which provides practical guidance and methods for the wide application of UPT in the field of meat thawing.

Efficient degradation of imidacloprid by surface discharge cold plasma: Mechanism of interaction between ROS and molecular structure and evaluation of residual toxicity.

Incorrect use of neonicotinoid pesticides poses a serious threat to human and pollinator health, as these substances are commonly present in bee products and even drinking water. To combat this threat, the study developed a new method of degrading the pesticide imidacloprid using surface discharge cold plasma oxidation technology. The study showed that this method achieved a very high efficiency of imidacloprid degradation of 91.4%. The main reactive oxygen species (H2O2, O3, ·OH, O2-, 1O2) effectively participated in the decomposition reaction of imidacloprid. Reactive oxygen species were more sensitive to the structure of the nitroimine group. Density functional theory (DFT) further explored the sites of reactive oxygen species attack on imidacloprid and revealed the process of energy change of attacking imidacloprid. In addition, a degradation pathway for imidacloprid was proposed, mainly involving reactive oxygen species chemisorption, a ring-opening intermediate, and complete cleavage of the nitroimine group structure. Model predictions indicated that acute oral and developmental toxicity were significantly reduced after cold plasma treatment, as confirmed by insect experiments. Animal experiments have shown that plasma treatment reduces imidacloprid damage to mice hippocampal tissue structure and inhibits the reduction of brain-derived neurotrophic factor content, thus revealing the detoxification mechanism of the body.

The combined effects of ultrasound and plasma-activated water on silkworm pupae:Physicochemical properties, microbiological diversity and ultrastructure.

A novel technique was proposed for processing silkworm pupae by combining plasma- activated water (PAW) with ultrasound (US). The microbial diversity and quality characteristics of the silkworm pupae were also evaluated. The results of the microbial diversity analysis indicated that PAW combined with US treatment significantly reduced the relative abundance of Streptococcaceae, Leuconostocaceae, and Acetobacteraceae from 32%, 18% and 16% to 27%, 11% and 11%, respectively. Microstructural analysis demonstrated that the collapse of the internal structure of chitin in silkworm pupae facilitated the release of nutrients and flavour compounds including fatty acids, water-soluble proteins (WSP), amino acids, phenolics, and volatile compounds. Furthermore, the increase in antioxidant capacity and the decrease in catalase activity and malondialdehyde content confirmed the mechanism of quality change. These findings provide new insights into the possible mechanism of PAW combined with US to improve the quality of edible insects.

Measurement of entropy production in living cells under an alternating electric field.

Entropy is a thermodynamic property toward equilibrium based on the dissipation of energy. Cells constitute such a thermodynamic system, in which entropy production is both inevitable and highly significant. Although the experimental measurement of entropy production in a cell is very difficult, a new method to accomplish this in living cells is reported herein. Through heating the sample by alternating electric fields and recording the heat flow from cells, the entropy production in two normal cell lines, MCF10A and HL-7702, and two cancerous cell lines, MDA-MB-231 and SMMC-7721, was measured and compared. The scaled electroinduced entropy production rate (SEEP) of cancer cells monotonically increases with electric field strength at 5-40 V/cm, while that of normal cells changes nonmonotonically with electric field strength, reaching a peak at 5-30 V/cm. For all cell lines, the cancerous-to-normal ratio of field-induced entropy production is clearly <1 in a large range of field strength from 5 to 25 V/cm. Therefore, this work presents an easy and effective strategy for experimentally investigating the thermodynamic properties of the cell, and gives deeper insight into the physical differences between normal and cancerous cells exposed to electric fields.

Effect of Electrohydrodynamic Drying on Drying Characteristics and Physicochemical Properties of Carrot.

This study investigates the effects of electrohydrodynamic (EHD) drying technology on the drying kinetics, microstructure, quality, and nutritional components of carrots, along with conducting experiments on EHD drying under different voltage gradients. The experimental results showed that EHD drying technology could significantly increase the drying rate and the effective moisture diffusion coefficient. Within a certain range, the drying rate was directly proportional to the voltage. When the range was exceeded, the increase in voltage had a minimal effect on the drying rate. In terms of quality, the EHD drying group's color, shrinkage rate, and rehydration performance were superior to the control group, and different voltages had no significant effect on the shrinkage rate and rehydration performance. The retention of carotenoids in the EHD drying group was 1.58 to 2 times that of the control group. EHD drying had a negative impact on the total phenolic content and vitamin A content of dried carrot slices. Based on the results of infrared spectroscopy and scanning electron microscopy (SEM), the dehydrated carrot slices showed wrinkling due to water loss, with numerous pores, a generally intact structure, and retained functional groups. EHD drying had a significant impact on the secondary structure of proteins, where an increase in voltage led to an increase in disordered structure, with a smaller proportion of disordered structure in the lower voltage group compared to the control group, and a similar proportion of disordered structure between the higher voltage group and the control group. Results from low-field nuclear magnetic resonance (NMR) showed that EHD drying could retain more bound water compared to the control group, with the best retention of cellular bound water at a voltage of 26 kV and the best retention of cellular immobilized water at a voltage of 38 kV, indicating the superiority of EHD drying in preserving cellular structure. This study provided a theoretical basis and experimental foundation for the application of electrohydrodynamic drying technology to carrot drying, and promoted the practical application of EHD drying technology.

Influence of electrohydrodynamics on the drying characteristics and physicochemical properties of garlic.

Electrohydrodynamic (EHD) drying, natural air drying (AD) and hot air drying (HAD) were used to comprehensively study the drying characteristics and physicochemical properties of garlic, and low-field nuclear magnetic resonance (NMR), infrared spectroscopy, scanning electron microscopy and other technologies were used as detection methods. In terms of drying characteristics, HAD has the largest effective diffusion coefficient of moisture and the shortest average drying time. EHD-treated garlic slices had the most attractive color, the highest rehydration rate, the most stable cell structure, the highest content of active ingredients, and the most stable protein secondary structure. Therefore, electrohydrodynamic drying is a promising garlic slice drying technology and provides an effective method for the large-scale production of high-quality garlic.

Electrohydrodynamic drying of citrus (Citrus sinensis L.) peel: Comparative evaluation on the physiochemical quality and volatile profiles.

Based on the concept of circular economy, citrus peel was considered a valuable source of bioactive compounds for high-value foods. Electrohydrodynamic (EHD) drying is a novel technology appropriated for the dehydration of heat-sensitive products such as citrus peel. In current work, EHD drying of citrus peel was performed based on alternating current (AC) or direct current (DC) sources at various voltage levels (9, 18, 27, 36, and 45 kV). The effect of EHD on drying characteristics, water retention capacity, enzyme inactivation, phytochemical contents (phenolic compounds and carotenoids), and volatile compounds of citrus peel were evaluated and compared. Results showed that the drying time in the AC electric field was shorter compared to DC electric field at the same applied voltages due to the polarization layer formed by unipolar charges. The applied voltage determined electric field strength as well as the degree of tissue collapse and cell membrane rupture. EHD elucidated the transformation and degradation of phytochemicals including phenolic compounds, carotenoids, and volatile composition in proportion to the applied voltage. The findings indicate that EHD drying with AC improves drying behaviors, inactivates enzymes, and retains the phytochemical properties of citrus peel.

Influence of electrohydrodynamics on the drying characteristics and volatile components of iron stick yam.

The drying characteristics, rehydration capacity, color, infrared spectra and volatile components of iron stick yam slices were investigated under different alternating current (AC) voltages (13, 17, 21 kV), hot air drying (HAD) (60 °C) and natural drying (AD) by electrohydrodynamic (EHD) drying and HAD experimental devices. The results showed that slices of iron stick yam dried the quickest with HAD, which also had the fastest drying rate; while drying the slices of iron stick yam with EHD led to a better rehydration capacity, higher brightness L* and whiteness, a more stable protein secondary structure, and a greater variety and content of volatile components compared with AD and HAD. These finding indicated that EHD is a more promising method for drying iron stick yam.

Effect of Electrohydrodynamic (EHD) on Drying Kinetics and Quality Characteristics of Shiitake Mushroom.

The effect of an electrohydrodynamic (EHD) drying system on the drying kinetics, microstructure and nutritional composition of shiitake mushrooms was studied. Shiitake mushroom slices were dried at 0, 18, 22, 26, 30 and 34 kV. The results showed that the drying rate, effective moisture diffusion coefficient and shrinkage of the EHD treatment group were significantly higher than those of the control group. The 34 kV treatment group had the highest drying rate (0.24 g W/g DM × h) and the highest effective moisture diffusion coefficient (1.01 × 10-10 m2/s), which were 6.75 and 7.41 times higher than those of the control group, respectively. The control group had the highest rehydration ratio (7.72) and showed unsatisfactory color performance. The scanning electron microscopy (SEM) results showed that compared with the control group, the surface of samples dried by EHD exhibited different degrees of encrustation, and the area of encrustation increased with increasing voltage. After analysis by Fourier transform infrared (FTIR) spectroscopy, it was found that the samples of both the EHD-treated and control groups had similar absorption peak positions, but the intensity of the absorption peak of the EHD-dried samples was greater. Compared with the control group, the shiitake mushroom slices dried by EHD had a higher protein content and polysaccharide content. The polysaccharide content in 22 kV treatment group was the highest (4.67 g/100 g), and the protein content in 26 kV and 34 kV treatment groups was the highest (17.0 g/100 g). This study provides an experimental and theoretical basis for an in-depth study of the drying kinetics of shiitake mushrooms and provides theoretical guidance and clues for the wider application of EHD drying technology.

Gene expression variation of Astragalus adsurgens Pall. through discharge plasma and its activated water.

To explore the effects of plasma-activated water (PAW) on gene expression, the combined treatment of PAW and discharge plasma on Astragalus adsurgens Pall seeds were performed, and then the gene expression of seedlings after treatmentwas analyzed at the molecular level. A needle array-plate dielectric-barrier discharge plasma was used to treat Astragalus adsurgens Pall seeds for 1, 2, and 3 h, and PAW was prepared at the same time to cultivate seeds. When the treatment time was 3 h, the survival rate of Plasma + PAW seedlings was only 9.2% of that of the CK. The Astragalus adsurgens Pall seedlings were analyzed using reactive oxygen species (ROS) and RNA-Seq. The ROS content of the seedlings in treatment group was significantly higher than that in the CK after 3 days of culture, that PAW cultivated can cause oxidative stress damage to Astragalus adsurgens Pall. The enzyme activity of the treated plant increased and the metabolic rate was accelerated. It helped to regulate the growth process of plants and improve the yield and quality of crops. This study discussed the gene expression of plasma and PAW induced Astragalus adsurgens Pall at the molecular level, and provided experimental data support for plasma and PAW treatment and selection of Astragalus adsurgens Pall.

Spectral characteristics on increasing hydrophilicity of Alfalfa seeds treated with alternating current corona discharge field.

In this study, corona discharge at atmospheric pressure under an alternating current (AC) field with a multi-needle plate electrode structure was used as mutagen to study the effects of corona discharge field on the hydrophilicity of alfalfa seeds, especially the effects of non-uniform electric field that is frequently ignored. Alfalfa seed were divided into two groups, one group was covered with 1 mm-thick polypropylene petri dish cover that can be approximated as a single factor effect of non-uniform electric field, the other group was directly irradiated by corona discharge field under different parameters. Fourier transform infrared spectroscopy (FTIR) and dual-index sequence analysis were used to detect and compare the spectra of alfalfa seed coats with different treatments. Results showed that the peaks of the seed coat at 2856 and 1727 cm-1 in the treatment group changed, indicating that hydrophobic wax, lipid, and cellulose may be cracked or degraded. Corona discharge field treatment can change the chemical structure of alfalfa seed coats, resulting in the changes in their hydrophilicity. The results of the apparent contact angle and scanning electron microscopy (SEM) and other experiments showed that the change in 19 kV non-covered treatment group was greatest, the petri dish cover can effectively reduce the physicochemical etching of ionic wind, the hydrophilicity of alfalfa seeds improved after corona discharge field treatment regardless of whether there is an ionic effect during irradiation, and the ionic wind has a greater influence on the hydrophilicity of alfalfa seeds than the non-uniform electric field. This study revealed the biological effect mechanism of corona discharge field from the perspective of spectral characteristics and provided experimental data support for the analysis of alfalfa seed surface modification and chemical structure after corona discharge field treatment.

Electrohydrodynamic Drying of Chinese Wolfberry in a Multiple Needle-to-Plate Electrode System.

In order to systematically and comprehensively investigate electrohydrodynamic (EHD) drying characteristics and mechanisms in a multiple needle-to-plate electrode system, drying experiments of Chinese wolfberry were conducted by blocking ionic wind and changing needle spacing in a multiple needle-to-plate electrode system. Drying characteristics, quality parameters, and the microstructure of Chinese wolfberry fruits were measured. Results show that ionic wind plays a very important role during the drying process. Drying rates of different needle spacing treatments are significantly higher than that of the control, and the drying rate decreases with the increase of needle spacing. Needle spacing has a great influence on the speed of ionic wind, rehydration rate, and polysaccharide contents. The effective moisture diffusion coefficient and the electrical conductivity disintegration index decreases with an increase in needle spacing. Ionic wind has a great influence on the effective moisture diffusion coefficient and the electrical conductivity disintegration index of Chinese wolfberry fruits. The microstructure of Chinese wolfberry fruits dried in an EHD system significantly changed. This study provides a theoretical basis and practical guidance for understanding characteristic parameters and mechanisms of EHD drying technology.

Using feature optimization-based support vector machine method to recognize the ß-hairpin motifs in enzymes.

ß-Hairpins in enzyme, a kind of special protein with catalytic functions, contain many binding sites which are essential for the functions of enzyme. With the increasing number of observed enzyme protein sequences, it is of especial importance to use bioinformatics techniques to quickly and accurately identify the ß-hairpin in enzyme protein for further advanced annotation of structure and function of enzyme. In this work, the proposed method was trained and tested on a non-redundant enzyme ß-hairpin database containing 2818 ß-hairpins and 1098 non-ß-hairpins. With 5-fold cross-validation on the training dataset, the overall accuracy of 90.08% and Matthew's correlation coefficient (Mcc) of 0.74 were obtained, while on the independent test dataset, the overall accuracy of 88.93% and Mcc of 0.76 were achieved. Furthermore, the method was validated on 845 ß-hairpins with ligand binding sites. With 5-fold cross-validation on the training dataset and independent test on the test dataset, the overall accuracies were 85.82% (Mcc of 0.71) and 84.78% (Mcc of 0.70), respectively. With an integration of mRMR feature selection and SVM algorithm, a reasonable high accuracy was achieved, indicating the method to be an effective tool for the further studies of ß-hairpins in enzymes structure. Additionally, as a novelty for function prediction of enzymes, ß-hairpins with ligand binding sites were predicted. Based on this work, a web server was constructed to predict ß-hairpin motifs in enzymes (http://202.207.29.251:8080/).

Identification of metal ion binding sites based on amino acid sequences.

The identification of metal ion binding sites is important for protein function annotation and the design of new drug molecules. This study presents an effective method of analyzing and identifying the binding residues of metal ions based solely on sequence information. Ten metal ions were extracted from the BioLip database: Zn2+, Cu2+, Fe2+, Fe3+, Ca2+, Mg2+, Mn2+, Na+, K+ and Co2+. The analysis showed that Zn2+, Cu2+, Fe2+, Fe3+, and Co2+ were sensitive to the conservation of amino acids at binding sites, and promising results can be achieved using the Position Weight Scoring Matrix algorithm, with an accuracy of over 79.9% and a Matthews correlation coefficient of over 0.6. The binding sites of other metals can also be accurately identified using the Support Vector Machine algorithm with multifeature parameters as input. In addition, we found that Ca2+ was insensitive to hydrophobicity and hydrophilicity information and Mn2+ was insensitive to polarization charge information. An online server was constructed based on the framework of the proposed method and is freely available at http://60.31.198.140:8081/metal/HomePage/HomePage.html.