Spinach-based RNA mimicking GFP in plant cells.

Spinach RNA-mimicking GFP (S-RMG) has been successfully used to monitor cellular RNAs including microRNAs in bacterium, yeast, and human cells. However, S-RMG has not been established in plants. In this study, we found that like bacterial, yeast, and human cellular tRNAs, plant tRNAs such as tRNALys can protect and/or stabilize the Spinach RNA aptamer interaction with the fluorophore DFHBI enabling detectable levels of green fluorescence to be emitted. The tRNALys-Spinach-tRNALys, once delivered into "chloroplast-free" onion epidermal cells can emit strong green fluorescence in the presence of DFHBI. Our results demonstrate for the first time that Spinach-based RNA visualization has the potential for in vivo monitoring of RNAs in plant cells.

Mitigating fungal contamination of cereals: The efficacy of microplasma-based far-UVC lamps against Aspergillus flavus and Fusarium graminearum.

Fungal contaminations of cereal grains are a profound food-safety and food-security concern worldwide, threatening consumers' and animals' health and causing enormous economic burdens. Because far-ultraviolet C (far-UVC) light at 222 nm has recently been shown to be human-safe, we investigated its efficacy as an alternative to thermal, chemical, and conventional 254 nm UVC anti-fungal treatments. Our microplasma-based far-UVC lamp system achieved a 5.21-log reduction in the conidia of Aspergillus flavus suspended in buffer with a dose of 1032.0 mJ/cm2, and a 5.11-log reduction of Fusarium graminearum conidia in suspension with a dose of 619.2 mJ/cm2. We further observed that far-UVC treatments could induce fungal-cell apoptosis, alter mitochondrial membrane potential, lead to the accumulation of intracellular reactive oxygen species, cause lipid peroxidation, and result in cell-membrane damage. The lamp system also exhibited a potent ability to inhibit the mycelial growth of both A. flavus and F. graminearum. On potato dextrose agar plates, such growth was completely inhibited after doses of 576.0 mJ/cm2 and 460.8 mJ/cm2, respectively. To test our approach's efficacy at decontaminating actual cereal grains, we designed a cubical 3D treatment chamber fitted with six lamps. At a dose of 780.0 mJ/cm2 on each side, the chamber achieved a 1.88-log reduction of A. flavus on dried yellow corn kernels and a 1.11-log reduction of F. graminearum on wheat grains, without significant moisture loss to either cereal type (p > 0.05). The treatment did not cause significant changes in the propensity of wheat grains to germinate in the week following treatment (p > 0.05). However, it increased the germination propensity of corn kernels by more than 71% in the same timeframe (p < 0.05). Collectively, our results demonstrate that 222 nm far-UVC radiation can effectively inactivate fungal growth in liquid, on solid surfaces, and on cereal grains. If scalable, its emergence as a safe, cost-effective alternative tool for reducing fungi-related post-harvest cereal losses could have important positive implications for the fight against world hunger and food insecurity.

Valorization of Food Waste: Utilizing Natural Porous Materials Derived from Pomelo-Peel Biomass to Develop Triboelectric Nanogenerators for Energy Harvesting and Self-Powered Sensing.

Food waste is an enormous challenge, with implications for the environment, society, and economy. Every year around the world, 1.3 billion tons of food are wasted or lost, and food waste-associated costs are around $2.6 trillion. Waste upcycling has been shown to mitigate these negative impacts. This study's optimized pomelo-peel biomass-derived porous material-based triboelectric nanogenerator (PP-TENG) had an open circuit voltage of 58 V and a peak power density of 254.8 mW/m2. As porous structures enable such triboelectric devices to respond sensitively to external mechanical stimuli, we tested our optimized PP-TENG's ability to serve as a self-powered sensor of biomechanical motions. As well as successfully harvesting sufficient mechanical energy to power light-emitting diodes and portable electronics, our PP-TENGs successfully monitored joint motions, neck movements, and gait patterns, suggesting their strong potential for use in healthcare monitoring and physical rehabilitation, among other applications. As such, the present work opens up various new possibilities for transforming a prolific type of food waste into value-added products and thus could enhance long-term sustainability while reducing such waste.

Intelligent films based on pectin, sodium alginate, cellulose nanocrystals, and anthocyanins for monitoring food freshness.

To address consumer-level food waste, and pollution from commercial plastics, we developed intelligent films using sodium alginate (SA), pectin (PC), cellulose nanocrystals (CNCs), and anthocyanins extracted from red cabbage (RCA). We also investigated two methods of reinforcing these films - cross-linking (CL), and the addition of CNCs. Both together and separately, these methods improved SA/PC films' mechanical properties and thermal stability. The optimal SA/PC/CNCs/RCA/CL films exhibited pH-dependent color-response properties and high water resistance. These were then tested as colorimetric freshness indicators for shrimp samples, both through seepage and the monitoring of volatile compounds. The colors of the indicators changed from lilac to dark green to greenish-yellow after storage at 25 °C for 72 h, whereas at 4 °C, they changed much more slowly over the same time period. This demonstrated the excellent potential of such films to reduce food waste by providing real-time warnings of pH variation resulting from spoilage.

Transcriptional and post-transcriptional regulation of RNAi-related gene expression during plant-virus interactions.

As sessile organisms, plants encounter diverse invasions from pathogens including viruses. To survive and thrive, plants have evolved multilayered defense mechanisms to combat virus infection. RNAi, also known as RNA silencing, is an across-kingdom innate immunity and gene regulatory machinery. Molecular framework and crucial roles of RNAi in antiviral defense have been well-characterized. However, it is largely unknown that how RNAi is transcriptionally regulated to initiate, maintain and enhance cellular silencing under normal or stress conditions. Recently, insights into the transcriptional and post-transcriptional regulation of RNAi-related genes in different physiological processes have been emerging. In this review, we integrate these new findings to provide updated views on how plants modulate RNAi machinery at the (post-) transcriptional level to respond to virus infection.

Linking calcium and RNAi signaling in plants.

The genetic link between calcium signaling and RNA interference (RNAi) has remained undiscovered until now. A new study shows that wound-triggered calcium flux acts as an initial messenger for priming RNAi for its role in plant antiviral defense. This paves the way to investigate plant development and response to (a)biotic stresses.

Hotspot siRNA Confers Plant Resistance against Viral Infection.

A hallmark of antiviral RNA interference (RNAi) is the production of viral small interfering RNA (vsiRNA). Profiling of vsiRNAs indicates that certain regions of viral RNA genome or transcribed viral RNA, dubbed vsiRNA hotspots, are more prone to RNAi-mediated cleavage for vsiRNA biogenesis. However, the biological relevance of hotspot vsiRNAs to the host innate defence against pathogens remains to be elucidated. Here, we show that direct targeting a hotspot by a synthetic vsiRNA confers host resistance to virus infection. Using Northern blotting and RNAseq, we obtained a profile of vsiRNAs of the African cassava mosaic virus (ACMV), a single-stranded DNA virus. Sense and anti-sense strands of small RNAs corresponding to a hotspot and a coldspot vsiRNA were synthesised. Co-inoculation of Nicotiana benthamiana with the double-stranded hotspot siRNA protected plants from ACMV infection, where viral DNA replication and accumulation of viral mRNA were undetectable. The sense or anti-sense strand of this hotspot vsiRNA, and the coldspot vsiRNA in both double-stranded and single-stranded formats possessed no activity in viral protection. We further demonstrated that the hotspot vsiRNA-mediated virus resistance had a threshold effect and required an active RDR6. These data show that hotspot vsiRNAs bear a functional significance on antiviral RNAi, suggesting that they may have the potential as an exogenous protection agent for controlling destructive viral diseases in plants.

Developing a unidirectionally permeable edible film based on ĸ-carrageenan and gelatin for visually detecting the freshness of grass carp fillets.

In this study, a unidirectionally permeable film (UPF, the inner layer is a hydrophilic film and outer layer is a hydrophobic film) was applied to the preservation of grass carp fillets for the first time. The hydrophilic film was made of ĸ-carrageenan, gelatin, and curcumin, which was a smart indicator for aquatic product's freshness detection. The hydrophobic film was made of zein. The optimum dosage of UPF were ĸ-carrageenan (1.5 % w/v), gelatin (1.03 % w/v), curcumin (60 mg/100 mL), zein (10.1 % w/v) and glycerol (0.29 % w/w) according to response surface methodology. The results showed that UPF had better mechanical strength and was more resistant to water than hydrophilic films (p < 0.05). Subsequently, UPF was implemented to pack and store fresh grass carp fillets at 4 ℃. With the increase of storage time, UPF detected the change of freshness and informed consumers through its color changes from yellow to red.

Mini review: Revisiting mobile RNA silencing in plants.

Non-cell autonomous RNA silencing can spread from cell to cell and over long-distances in animals and plants. This process is genetically determined and requires mobile RNA signals. Genetic requirement and molecular nature of the mobile signals for non-cell-autonomous RNA silencing were intensively investigated in past few decades. No consensus dogma for mobile silencing can be reached in plants, yet published data are sometimes inconsistent and controversial. Thus, the genetic requirements and molecular signals involved in plant mobile silencing are still poorly understood. This article revisits our present understanding of intercellular and systemic non-cell autonomous RNA silencing, and summarises current debates on RNA signals for mobile silencing. In particular, we discuss new evidence on siRNA mobility, a DCL2-dependent genetic network for mobile silencing and its potential biological relevance as well as 22 nt siRNA being a mobile signal for non-cell-autonomous silencing in both Arabidopsis and Nicotiana benthamiana. This sets up a new trend in unravelling genetic components and small RNA signal molecules for mobile silencing in (across) plants and other organisms of different kingdoms. Finally we raise several outstanding questions that need to be addressed in future plant silencing research.