Deciphering the mechanism of E3 ubiquitin ligases in plant responses to abiotic and biotic stresses and perspectives on PROTACs for crop resistance.

With global climate change, it is essential to find strategies to make crops more resistant to different stresses and guarantee food security worldwide. E3 ubiquitin ligases are critical regulatory elements that are gaining importance due to their role in selecting proteins for degradation in the ubiquitin-proteasome proteolysis pathway. The role of E3 Ub ligases has been demonstrated in numerous cellular processes in plants responding to biotic and abiotic stresses. E3 Ub ligases are considered a class of proteins that are difficult to control by conventional inhibitors, as they lack a standard active site with pocket, and their biological activity is mainly due to protein-protein interactions with transient conformational changes. Proteolysis-targeted chimeras (PROTACs) are a new class of heterobifunctional molecules that have emerged in recent years as relevant alternatives for incurable human diseases like cancer because they can target recalcitrant proteins for destruction. PROTACs interact with the ubiquitin-proteasome system, principally the E3 Ub ligase in the cell, and facilitate proteasome turnover of the proteins of interest. PROTAC strategies harness the essential functions of E3 Ub ligases for proteasomal degradation of proteins involved in dysfunction. This review examines critical advances in E3 Ub ligase research in plant responses to biotic and abiotic stresses. It highlights how PROTACs can be applied to target proteins involved in plant stress response to mitigate pathogenic agents and environmental adversities.

Recent advances in the multifaceted functions of Cys2/His2-type zinc finger proteins in plant growth, development, and stress responses.

Recent research has highlighted the importance of Cys2/His2-type zinc finger proteins (C2H2-ZFPs) in plant growth and in responses to various stressors, and the complex structures of C2H2-ZFP networks and the molecular mechanisms underlying their responses to stress have received considerable attention. Here, we review the structural characteristics and classification of C2H2-ZFPs, and consider recent research advances in their functions. We systematically introduce the roles of these proteins across diverse aspects of plant biology, encompassing growth and development, and responses to biotic and abiotic stresses, and in doing so hope to lay the foundations for further functional studies of C2H2-ZFPs in the future.

The proteome of Penicillium expansum during infection of postharvest apple is revealed using Label-Free and Parallel Reaction Monitoring(PRM)Techniques.

Penicillium expansum is the main pathogen in the postharvest storage of apples. Penicilliosis caused by P. expansum infection not only seriously affects the appearance and quality of fruits, but also the secondary metabolite Patulin (PAT) can cause harm to human health. Until now, little attention has been paid to the molecular mechanism of P. expansum infecting apples. Studying its molecular mechanism can help us better prevent and control apple postharvest blue mold. In this present investigation, we will use Label-Free technology to perform proteomic sequencing on apple samples at key time points of P. expansum infection, explore and screen key proteins and metabolic pathways during infection, and use Parallel Reaction Monitoring (PRM) technology to thoroughly validate proteomic data. The infection of P. expansum activates the MAPK signaling pathway, plant-pathogen interaction metabolic pathway and phenylpropanoid biosynthesis pathway of apple, participates in the regulation of ROS generation and oxidative stress process, promotes the synthesis of lignin and flavonoids, and the synthesis of Pathogenesis-Related Protein helps apple directly defend against P. expansum infection. This study provides the foundation for relevant postharvest control strategies, paving the way for further exploration of the proteome of pathogens infecting fruit and vegetables. SIGNIFICANCE: Proteins are macromolecules essential to the life of organisms, as they participate in the function and structure of cells. Proteomics technology is currently one of the important means to study the the response mechanism of pathogenic bacteria to plant infection, which can reveal the essence of physiological and pathological processes and help to clarify the possible relationship between protein abundance and plant stress. The present study essentially uses recent proteome analysis technology, namely label-free and PRM techniques, and lays the foundations for studying the of the infection response between P. expansum and apples. In particular, it provides a broad perspective on the molecular mechanism of P. expansum in the early stage of apple infection through detailed functional exploration and verification of associated proteins. Thus, it provides a theoretical basis for preventing and treating apple postharvest blue mold.

A First Expression, Purification and Characterization of Endo-ß-1,3-Glucanase from Penicillium expansum.

ß-1,3-glucanase plays an important role in the biodegradation, reconstruction, and development of ß-1,3-glucan. An endo-ß-1,3-glucanase which was encoded by PeBgl1 was expressed, purified and characterized from Penicillium expansum for the first time. The PeBgl1 gene was amplified and transformed into the competent cells of E. coli Rosetta strain with the help of the pET-30a cloning vector. The recombinant protein PeBgl1 was expressed successfully at the induction conditions of 0.8 mmol/L IPTG at 16 °C for 16 h and then was purified by nickel ion affinity chromatography. The optimum reaction temperature of PeBgl1 was 55 °C and it had maximal activity at pH 6.0 according to the enzymatic analysis. Na2HPO4-NaH2PO4 buffer (pH 6.0) and NaCl have inhibitory and enhancing effects on the enzyme activities, respectively. SDS, TritonX-100 and some metal ions (Mg2+, Ca2+, Ba2+, Cu2+, and Zn2+) have an inhibitory effect on the enzyme activity. The results showed that PeBgl1 protein has good enzyme activity at 50-60 °C and at pH 5.0-9.0, and it is not a metal dependent enzyme, which makes it robust for storage and transportation, ultimately holding great promise in green biotechnology and biorefining.

Overexpression of the SDR gene improves the ability of Meyerozyma guilliermondii to degrade patulin in pears and juices.

The intracellular enzymes of antagonistic yeast are effective in controlling patulin (PAT) contamination. However, countless enzymes that have been revealed remain functionally uncharacterized. The study built on previous transcriptomic data obtained by our research group to amplify and express a gene encoding a short-chain dehydrogenase/reductase (SDR) in Meyerozyma guilliermondii. Overexpression of SDR increased the tolerance of M. guilliermondii to PAT and the ability to degrade PAT of the intracellular enzymes. Furthermore, MgSDR-overexpressed M. guilliermondii showed higher PAT degradation in juices (apple and peach) and controlled the blue mold of pears at 20 °C and 4 °C while significantly reduced the content of PAT and the biomass of Penicillium expansum in decayed tissues than wild-type M. guilliermondii. This study provides theoretical references for the subsequent heterologous expression, formulation, and application of the SDR protein from M. guilliermondii and contributes to elucidating the PAT degradation mechanism of antagonistic yeasts.

Recent advances in Penicillium expansum infection mechanisms and current methods in controlling P. expansum in postharvest apples.

One of the most significant challenges associated with postharvest apple deterioration is the blue mold caused by Penicillium expansum, which leads to considerable economic losses to apple production industries. Apple fruits are susceptible to mold infection owing to their high nutrient and water content, and current physical control methods can delay but cannot completely inhibit P. expansum growth. Biological control methods present promising alternatives; however, they are not always cost effective and have application restrictions. P. expansum infection not only enhances disease pathogenicity, but also inhibits the expression of host-related defense genes. The implementation of new ways to investigate and control P. expansum are expected with the advent of omics technology. Advances in these techniques, together with molecular biology approaches such as targeted gene deletion and whole genome sequencing, will lead to a better understanding of the P. expansum infectious machinery. Here, we review the progress of research on the blue mold disease caused by P. expansum in apples, including physiological and molecular infection mechanisms, as well as various methods to control this common plant pathogen.

Study on the biocontrol effect and physiological mechanism of Hannaella sinensis on the blue mold decay of apples.

Blue mold decay is a major postharvest disease of apples, causing considerable losses to the apple industry. In the early stage of this research, an antagonistic yeast, Hannaella sinensis, with a good control effect on the blue mold of apples, was selected. On this basis, the main purpose of this work was to study the biocontrol effect of H. sinensis on the blue mold of apples and the mechanisms involved. The results showed that H. sinensis could effectively control the blue mold decay of apples, reduce the rot rate and diameter, and the antagonistic effect strengthened with the increase of H. sinensis concentration (1 × 108 cells/mL). Further in vitro experiments proved that H. sinensis could significantly inhibit the spore germination and germ tube length of P. expansum. In addition, stable colonization of H. sinensis on apple wounds and surfaces confirmed the environmental adaptability and the ability to compete with other microbiota for nutrition and space. Moreover, H. sinensis induced the activities of resistance-related enzymes such as polyphenol oxidase (PPO), peroxidase (POD), ascorbate peroxidase (APX), superoxide dismutase (SOD), and phenylalanine ammonia-lyase (PAL) in apples and the content of the coding genes corresponding to these enzymes was also higher than that of the control group. Our results indicate that H. sinensis treatment could induce the disease resistance of apples. In summary, H. sinensis served as a promising antagonistic yeast for the prevention and treatment of postharvest blue mold decay of apples.

Unveiling ochratoxin a controlling and biodetoxification molecular mechanisms: Opportunities to secure foodstuffs from OTA contamination.

Anarchic growth of ochratoxin A (OTA) producing fungi during crop production, prolonged storage, and processing results in OTA contamination in foodstuffs. OTA in food exacerbates the risk of health and economic problems for consumers and farmers worldwide. Although the toxic effects of OTA on human health have not been well established, comprehensive preventive and remedial measures will be essential to eliminate OTA from foodstuffs. Strict regulations, controlling OTA at pre- or post-harvest stage, and decontamination of OTA have been adopted to prevent human and animal OTA exposure. Biological control of OTA and bio-decontamination are the most promising strategies due to their safety, specificity and nutritional value. This review addresses the current understanding of OTA biodegradation mechanisms and recent developments in OTA control and bio-decontamination strategies. Additionally, this review analyses the strength and weaknesses of different OTA control methods and the contemporary approaches to enhance the efficiency of biocontrol agents. Overall, this review will support the implementation of new strategies to effectively control OTA in food sectors. Further studies on efficacy-related issues, production issues and cost-effectiveness of OTA biocontrol are to be carried out to improve the knowledge, develop improved delivery technologies and safeguard the durability of OTA biocontrol approaches.