Enhanced laccase production by mutagenized Myrothecium verrucaria using corn stover as a carbon source and its potential in the degradation of 2-chlorophen.

Chlorophenols are widely used in industry and are known environmental pollutants. The degradation of chlorophenols is important for environmental remediation. In this study, we evaluated the biodegradation of 2-chlorophenol using crude laccase produced by Myrothecium verrucaria. Atmospheric and room temperature plasma technology was used to increase laccase production. The culture conditions of the M-6 mutant were optimized. Our results showed that corn stover could replace glucose as a carbon source and promote laccase production. The maximum laccase activity of 30.08 U/mL was achieved after optimization, which was a 19.04-fold increase. The biodegradation rate of 2-chlorophenol using crude laccase was 97.13%, a positive correlation was determined between laccase activity and degradation rate. The toxicity of 2-CP was substantially reduced after degradation by laccase solution. Our findings show the feasibility of the use of corn stover in laccase production by M. verrucaria mutant and the subsequent biodegradation of 2-chlorophenol using crude laccase.

Flexible Wearables for Plants.

The excellent stretchability and biocompatibility of flexible sensors have inspired an emerging field of plant wearables, which enable intimate contact with the plants to continuously monitor the growth status and localized microclimate in real-time. Plant flexible wearables provide a promising platform for the development of plant phenotype and the construction of intelligent agriculture via monitoring and regulating the critical physiological parameters and microclimate of plants. Here, the emerging applications of plant flexible wearables together with their pros and cons from four aspects, including physiological indicators, surrounding environment, crop quality, and active control of growth, are highlighted. Self-powered energy supply systems and signal transmission mechanisms are also elucidated. Furthermore, the future opportunities and challenges of plant wearables are discussed in detail.

High-Throughput and Accurate Determination of Transgene Copy Number and Zygosity in Transgenic Maize: From DNA Extraction to Data Analysis.

It is vital to develop high-throughput methods to determine transgene copy numbers initially and zygosity during subsequent breeding. In this study, the target sequence of the previously reported endogenous reference gene hmg was analyzed using 633 maize inbred lines, and two SNPs were observed. These SNPs significantly increased the PCR efficiency, while the newly developed hmg gene assay (hmg-taq-F2/R2) excluding these SNPs reduced the efficiency into normal ranges. The TaqMan amplification efficiency of bar and hmg with newly developed primers was calculated as 0.993 and 1.000, respectively. The inter-assay coefficient of variation (CV) values for the bar and hmg genes varied from 1.18 to 2.94%. The copy numbers of the transgene bar using new TaqMan assays were identical to those using dPCR. Significantly, the precision of one repetition reached 96.7% of that of three repetitions of single-copy plants analyzed by simple random sampling, and the actual accuracy reached 95.8%, confirmed by T1 and T2 progeny. With the high-throughput DNA extraction and automated data analysis procedures developed in this study, nearly 2700 samples could be analyzed within eight hours by two persons. The combined results suggested that the new hmg gene assay developed here could be a universal maize reference gene system, and the new assay has high throughput and high accuracy for large-scale screening of maize varieties around the world.

Identifying yield-related genes in maize based on ear trait plasticity.

BACKGROUND: Phenotypic plasticity is defined as the phenotypic variation of a trait when an organism is exposed to different environments, and it is closely related to genotype. Exploring the genetic basis behind the phenotypic plasticity of ear traits in maize is critical to achieve climate-stable yields, particularly given the unpredictable effects of climate change. Performing genetic field studies in maize requires development of a fast, reliable, and automated system for phenotyping large numbers of samples. RESULTS: Here, we develop MAIZTRO as an automated maize ear phenotyping platform for high-throughput measurements in the field. Using this platform, we analyze 15 common ear phenotypes and their phenotypic plasticity variation in 3819 transgenic maize inbred lines targeting 717 genes, along with the wild type lines of the same genetic background, in multiple field environments in two consecutive years. Kernel number is chosen as the primary target phenotype because it is a key trait for improving the grain yield and ensuring yield stability. We analyze the phenotypic plasticity of the transgenic lines in different environments and identify 34 candidate genes that may regulate the phenotypic plasticity of kernel number. CONCLUSIONS: Our results suggest that as an integrated and efficient phenotyping platform for measuring maize ear traits, MAIZTRO can help to explore new traits that are important for improving and stabilizing the yield. This study indicates that genes and alleles related with ear trait plasticity can be identified using transgenic maize inbred populations.

Gallium-Based Liquid Metal Materials for Antimicrobial Applications.

The hazards caused by drug-resistant bacteria are rocketing along with the indiscriminate use of antibiotics. The development of new non-antibiotic antibacterial drugs is urgent. The excellent biocompatibility and diverse multifunctionalities of liquid metal have stimulated the studies of antibacterial application. Several gallium-based antimicrobial agents have been developed based on the mechanism that gallium (a type of liquid metal) ions disorder the normal metabolism of iron ions. Other emerging strategies, such as physical sterilization by directly using LM microparticles to destroy the biofilm of bacteria or thermal destruction via infrared laser irradiation, are gaining increasing attention. Different from traditional antibacterial agents of gallium compounds, the pronounced property of gallium-based liquid metal materials would bring innovation to the antibacterial field. Here, LM-based antimicrobial mechanisms, including iron metabolism disorder, production of reactive oxygen species, thermal injury, and mechanical destruction, are highlighted. Antimicrobial applications of LM-based materials are summarized and divided into five categories, including liquid metal motors, antibacterial fabrics, magnetic field-responsive microparticles, liquid metal films, and liquid metal polymer composites. In addition, future opportunities and challenges towards the development and application of LM-based antimicrobial materials are presented.

Enhanced Production of Two Bioactive Isoflavone Aglycones in Astragalus membranaceus Hairy Root Cultures by Combining Deglycosylation and Elicitation of Immobilized Edible Aspergillus niger.

A cocultivation system of Astragalus membranaceus hairy root cultures (AMHRCs) and immobilized food-grade fungi was established for the enhanced production of calycosin (CA) and formononetin (FO). The highest accumulations of CA (730.88 ± 63.72 µg/g DW) and FO (1119.42 ± 95.85 µg/g DW) were achieved in 34 day-old AMHRCs cocultured with immobilized A. niger (IAN) for 54 h, which were 7.72- and 18.78-fold higher than CA and FO in nontreated control, respectively. IAN deglycosylation could promote the formation of CA and FO by conversion of their glycoside precursors. IAN elicitation could intensify the generation of endogenous signal molecules involved in plant defense response, which contributed to the significantly up-regulated expression of genes in CA and FO biosynthetic pathway. Overall, the coupled culture of IAN and AMHRCs offered a promising and effective in vitro approach to enhance the production of two health-promoting isoflavone aglycones for possible nutraceutical and pharmaceutical uses.

Preparation and applications of Arabidopsis thaliana guard cell protoplasts.

• Guard cells play an important role in the physiology and development of plants. The genetic resources available for Arabidopsis thaliana make it the most favorable plant species for the study of guard cell processes, but it is not easy to isolate highly purified preparations of large numbers of guard cells from this species. Here, we describe methods for isolation of both guard cell and mesophyll cell protoplasts from A. thaliana and their use in the study of unique biochemical and cellular properties of these cell types. • Protocols developed for large- and small-scale preparation of guard cell protoplasts and mesophyll cell protoplasts are described, followed by specific examples of their use in electrophysiological, biochemical and molecular approaches such as patch clamping, enzyme assays, and reverse-transcription polymerase chain reaction. • The protocols described yield millions of highly purified, viable guard cell protoplasts and mesophyll cell protoplasts from A. thaliana. These protoplasts have been used successfully in the study of ion channel properties, assay of ABA activation in phospholipase D activity and comparisons of gene and protein expression levels. • These techniques make it possible to elucidate electrophysiological, biochemical and molecular genetic pathways of guard cell function.

[Effects of hypertonic saline on erythrocyte adherence function and bacterial infection of hemorrhagic shock rabbits].

AIM: To explore the effect of hypertonic saline on the erythrocyte adherence function and bacterial infection of hemorrhagic shock rabbits. METHODS: 60 Japanese rabbits were randomly divided into 6 groups, 10 for each group. Artery catheterization and heparin were given to the rabbits in group 1 (sham shock group). Hemorrhagic shock model was set up by bleeding resulting from carotid artery catheter in group 2 (normal saline group )and group 3 (hypertonic saline group). 30 minutes after shock, the rabbits in group 1 and group 2 were treated with normal saline and balanced salt solution containing 1 x 10(9)/kg E.coli, respectively. And the rabbits in group 3 were treated with 75 g/L NaCl solution and balanced salt solution containing 1 x 10(9)/kg E.coli. Then the survival rates of the rabbits in group 1-3 were observed. Rabbits in group 4-6 were same treatment as received, group 1-3, respectively, except that there was no E.coli in balanced salt solution. The erythrocyte immune adherence function of rabbits in group 4-6 were detected 5 hours after shock by RBC-C3bR and RBC-IC rosette forming assays. RESULTS: The survival rate of rabbits in hypertonic saline group was significantly higher than that in normal saline group. The RBC-C3bR rosette forming rate of the normal saline treated rabbits were pronouncedly decreased, while RBC-IC rosette forming rate was notably elevated, as compared with those of either sham shock group or hypertonic saline group(P<0.01). Hypertonic saline markedly increased RBC C3bR rosette forming rate. CONCLUSION: The above findings suggest that hypertonic salt solution can remarkably improve the depressed erythrocyte immune adherence function and enhance the rabbit's resistance to E.coli challenge after hemorrhagic shock.