The realm of antimicrobial proteins in plants is extensive but remains relatively uncharted. Understanding the mechanisms underlying the action of plant antifungal proteins (AFPs) holds promise for antifungal strategies. This study aimed to bridge this knowledge gap by comprehensively screening Arabidopsis thaliana species to identify novel AFPs. Using MALDI-TOF analysis, we identified a member of the TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR1 (TCP) family of transcription factors as a novel AFP, A. thaliana TCP21 (AtTCP21; accession number NP_196450). Bacterially purified recombinant AtTCP21 inhibited the growth of various pathogenic fungal cells. AtTCP21 was more potent than melittin, a well-known AFP, in combating Colletotrichum gloeosporioides. Growth inhibition assays against various fungal pathogens and yeasts confirmed the pH-dependent antimicrobial activity of AtTCP21. Without inducing any membrane alterations, AtTCP21 penetrates the fungal cell wall and membrane, where it instigates a repressive milieu for fungal cell growth by generating intracellular reactive oxygen species and mitochondrial superoxides; resulting in morphological changes and apoptosis. Our findings demonstrate the redox-regulating effects of AtTCP21 and point to its potential as an antimicrobial agent.

Although considerable scientific research data is available for sepsis and cytokine storm syndrome, there is a need to develop new treatments or drugs for sepsis management. Antimicrobial peptides (AMPs) possess anti-bacterial and anti-inflammatory activity, neutralizing toxins such as lipopolysaccharides (LPS, endotoxin). Most AMPs have been designed as a substitute for conventional antibiotics, which kill drug-resistant pathogens. The present study aimed to determine the anti-inflammatory potential of 10 designed XIW (X: lysine, arginine, or glutamic acid) α-helical peptides in macrophages and a mouse model in the presence of LPS. Among them, WIKE-14, a peptide with a helix-to-helix structure, having the 12th amino acid substituted with glutamic acid, suppressed pro-inflammatory cytokines in RAW 264.7 macrophages. This reaction was mediated by the inhibition of the binding between LPS and macrophages. In addition, the WIKE-14 peptide exhibited a potent anti-inflammatory activity in mice ears and lungs inflamed using LPS. Thus, our results may provide useful insights for the development of anti-sepsis agents via the sequence and structure information of the WIKE-14 peptide.

Cotton (Gossypium hirsutum L.) is grown worldwide for its natural hollow fibers and is used as cattle feed. Living modified (LM) cotton is not cultivated in South Korea and must be imported for food, feed, and processing. From 2009 to 2013, the Ministry of Environment (MOE) and the National Institute of Ecology (NIE) conducted a natural environment monitoring and post-management initiative for living modified organisms (LMOs) in some areas to reduce the likelihood of harmful effects caused by unintentionally discharged LMOs during transportation and use. In this study, we adopted a new strategy to identify unintentionally released LM cotton plants nationwide from 2014 to 2018. A total of 451 suspicious cotton samples were collected from 3921 survey sites. Among them, we identified 255 LM cotton plants, of which approximately 72.2% had transgenic herbicide and insecticide traits. The majority of the samples were collected from the roadside along transportation routes and from stockbreeding farms. This study establishes an LMO safety management system to efficiently maintain conservation efforts in South Korea. Our findings suggest that these efforts may play a key role in safely transporting, using, and managing approved LMOs, as well as in regulating unintentionally released LMOs, in order to preserve the natural ecosystem of South Korea.

The unintentional release of living modified organisms (LMOs) into natural environments poses potential ecological risks, particularly in terms of gene flow and biodiversity loss. Since 2009, the Ministry of Environment (MOE) in South Korea has conducted an extensive monitoring project to detect and manage LMOs, with a primary focus on LM canola. This study evaluates the outcomes of the LMO monitoring project over the past 15 years (2009–2023), analyzing the distribution, persistence, and management of LM canola across various environments. Our findings reveal that LM canola predominantly proliferates along roadsides, with occasional occurrences at festival and planting sites. Out of 10,571 monitored sites, 4326 suspicious samples were collected, with a significant increase observed in 2017, underscoring the ongoing risk of accidental releases. This study highlights the critical role of specific environments in the spread of LM canola, and assesses the effectiveness of post-management strategies in controlling these populations. The National Institute of Ecology (NIE) has developed and implemented advanced monitoring protocols and post-management systems tailored to the characteristics of the monitoring sites and the nature of the LMOs. These efforts have been effective in controlling the spread of LM canola, thereby helping to preserve the biodiversity of South Korea’s natural environments. In conclusion, the proactive and adaptive strategies employed by the NIE are essential for mitigating the ecological risks associated with LMOs. Our study emphasizes the importance of on-going vigilance and the continuous refinement of monitoring and management practices to safeguard biodiversity and ecosystems.

Since 2009, the Ministry of Environment and the National Institute of Ecology in South Korea have been conducting a living modified organism (LMO) monitoring and post-management project in natural environments to prevent the unintentional release and spread of LMOs to natural ecosystems. The project surveyed six administrative districts of South Korea from 2009 to 2013 and collected 1960 LMO suspicious samples from 1850 monitoring sites. As a result, 113 LMOs were identified at 65 sites and removed for post-management. An analysis of the five-year LMO monitoring results showed that LMOs were re-collected in 38.4% of the 65 areas where they were initially collected. This result led to the establishment of a new LMO management system in 2014, with priority given to areas where LMOs had been re-collected twice or more within the last five years. Intensive surveys and post-management were conducted four times a year in these priority management areas. The results confirmed that the novel management system for LMO priority areas effectively prevented the continuous collection of LMOs in the same areas. In conclusion, establishing a safety management system for priority management areas, intensive surveys, and post-management efforts are crucial for protecting natural ecosystems from the putative risks of unintentionally released LMOs.

Various proteins introduced into living modified organism (LMO) crops function in plant defense mechanisms against target insect pests or herbicides. This study analyzed the antifungal effects of an introduced LMO protein, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium sp. strain CP4 (CP4-EPSPS). Pure recombinant CP4-EPSPS protein, expressed in Escherichia coli, inhibited the growth of human and plant fungal pathogens (Candida albicans, C. tropicalis, C. krusei, Colletotrichum gloeosporioides, Fusarium solani, F. graminearum, and Trichoderma virens), at minimum inhibitory concentrations (MICs) that ranged from 62.5 to 250 µg/mL. It inhibited fungal spore germination as well as cell proliferation on C. gloeosporioides. Rhodamine-labeled CP4-EPSPS accumulated on the fungal cell wall and within intracellular cytosol. In addition, the protein induced uptake of SYTOX Green into cells, but not into intracellular mitochondrial reactive oxygen species (ROS), indicating that its antifungal action was due to inducing the permeability of the fungal cell wall. Its antifungal action showed cell surface damage, as observed from fungal cell morphology. This study provided information on the effects of the LMO protein, EPSPS, on fungal growth.