Hydrogen sulphide regulates the growth of tomato root cells by affecting cell wall biosynthesis under CuO NPs stress

Copper oxide nanoparticles (CuO NPs) show strong nano-toxic effects on organisms. Hydrogen sulphide (H₂ S) plays a pivotal role in plant response to abiotic stress. In this study, we examine the crucial role of the cell wall as regulated by H₂ S in response to CuO NPs stress. The digestion method was employed to determine Cu content using atomic absorption spectrometry. The TraKine pro-tubulin staining kit was used to investigate the microtubule cytoskeleton using confocal laser-scanning microscopy. Cell wall component analysis utilized the ICS-3000 HPLC system. Application of H₂ S reduced growth inhibition caused by CuO NPs. Furthermore, most of the CuO NPs accumulates in roots, indicating a low transfer rate, and H₂ S significantly decreased CuO NPs content in roots, leaves and stems. Subcellular distribution analysis implied most Cu accumulated in root cell walls, and that H₂ S reduced the content of Cu in root cell walls. Cortical microtubules in the plasma membrane, guide cell wall biosynthesis. H₂ S obviously alleviated microtubule cytoskeleton disorders caused by CuO NPs. In addition, the content of cellulose, hemicellulose, pectin and other monosaccharides in root cell walls was reduced by CuO NPs treatment. H₂ S enhanced the monosaccharide and polysaccharide contents compared with that after CuO NPs treatment. In conclusion, H₂ S regulates cell wall development in response to CuO NPs stress by stabilizing microtubules. H₂ S affected Cu distribution and alleviated growth inhibition of tomato seedlings. The research results provide a theoretical basis for further study of nano-toxicity regulation in plants.