Comprehensive analysis of hepatocellular carcinoma 3D models revealed enhanced penetrative siRNA delivery by a nanodiamonds compared to liposomes. Nanodiamonds were able to improve siRNA's gene knockdown and anti-cancer effects in 3D tumor models.
The role of NADPH oxidases (NOXs) in pathogenesis and development in the Curvularia leaf spot (CuLS) agent Curvularia lunata remains poorly understood. In this study, we identified C. lunata ClNOX2, which localized to the plasma membrane and was responsible for reactive oxygen species (ROS) generation. Scavenging the ROS production inhibited the conidial germination and appressorial formation. The ClNOX2 and ClBRN1 deletion mutants were defective in 1,8-dihydroxynaphthalene (DHN) melanin accumulation, appressorial formation, and cellulase synthesis and exhibited lower virulence. However, disruption of the ClNOX2 and ClBRN1 genes facilitated hyphal growth, enhanced stress adaptation to cell wall-disrupting agents, and promoted developmental processes such as conidiation, conidial germination, and pseudothecium and ascus formation. Interestingly, loss of ClM1, the cell wall integrity (CWI) MAPK gene in C. lunata led to similar morphology and pathogenicity phenotypes with ClNOX2 and ClBRN1 deletion mutants, like abnormal conidia, fewer appressoria, less melanin, increased hyphal growth and enhanced tolerance to congo red (CR). These results indicated that the ClNOX2 gene plays an important role in C. lunata development and virulence via regulating intracellular DHN melanin biosynthesis. RT-qPCR revealed that the ClNOX2-related ROS signaling pathway and ClM1-mediated CWI signaling pathway are cross-linked in regulating DHN melanin biosynthesis. Our findings provide new insights into how ClNOX2 participates in the pathogenesis and development in hemibiotrophic plant fungal pathogens.
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